WO2025255452A2 - Molécules de liaison à l'antigène qui se lient à des particules d'aav et leurs utilisations - Google Patents

Molécules de liaison à l'antigène qui se lient à des particules d'aav et leurs utilisations

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Publication number
WO2025255452A2
WO2025255452A2 PCT/US2025/032630 US2025032630W WO2025255452A2 WO 2025255452 A2 WO2025255452 A2 WO 2025255452A2 US 2025032630 W US2025032630 W US 2025032630W WO 2025255452 A2 WO2025255452 A2 WO 2025255452A2
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seq
amino acid
acid sequence
abd1
abd2
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WO2025255452A3 (fr
Inventor
Christos Kyratsous
Andrew J. Murphy
Sven MOLLER-TANK
Yang Shen
Tri Nguyen
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Regeneron Pharmaceuticals Inc
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Regeneron Pharmaceuticals Inc
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Publication of WO2025255452A2 publication Critical patent/WO2025255452A2/fr
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Pending legal-status Critical Current
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/081—DNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2851—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the lectin superfamily, e.g. CD23, CD72
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2881—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD71
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2299/00—Coordinates from 3D structures of peptides, e.g. proteins or enzymes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/35—Valency
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/54—F(ab')2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/55—Fab or Fab'
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/64—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising a combination of variable region and constant region components
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/77—Internalization into the cell
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

Definitions

  • the instant application contains a Sequence Listing which has been submitted electronically in XML file format and is hereby incorporated by reference in its entirety. Said XML copy, created on May 30, 2025, is named 250298_000954_SL.xml and is 1,536,977 bytes in size. FIELD OF THE DISCLOSURE [0003] The present disclosure relates to antigen-binding molecules, including antibodies or antigen-binding fragments thereof, that bind to a capsid of an adeno-associated virus (AAV) particle.
  • AAV adeno-associated virus
  • multispecific antigen-binding molecules such as multispecific antibodies, e.g., bispecific antibodies, or antigen-binding fragments thereof, that bind to a capsid of an AAV and/or a molecule on a cell surface, as well as related molecular complexes and pharmaceutical compositions.
  • Methods for using the antibodies (e.g., multispecific antibodies) disclosed herein, molecular complexes and/or pharmaceutical composition are also provided.
  • AAV vector-mediated gene delivery to different cell types also varies greatly.
  • One possible mechanism for inefficient AAV transduction of certain cells may be a lack of cellular receptor(s) to mediate virus binding and entry.
  • the present disclosure provides antibodies that bind to a capsid of an adeno-associated virus (AAV) particle, and multispecific antibodies such as, for example, bispecific antibodies, which can bind both to the capsid of an AAV particle and a molecule (i.e., a target molecule) on a cell surface, and methods of use thereof.
  • AAV adeno-associated virus
  • multispecific antibodies such as, for example, bispecific antibodies, which can bind both to the capsid of an AAV particle and a molecule (i.e., a target molecule) on a cell surface
  • the antibodies described herein can bridge the virus to target cell surfaces that are not naturally targeted by the virus, thereby redirecting virus delivery and transduction.
  • a multispecific antibody or a multispecific antigen- binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a second heavy chain region of a second Fab (“Fab2”) operably linked to a second Fc domain (“Fc2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); and d) a fourth polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form Fab2, wherein Fab2 comprises a second antigen-binding domain (“ABD2”); wherein e) ABD
  • ABD1 binds to the capsid of the AAV particle and ABD2 binds to the molecule on the cell surface.
  • ABD2 binds to the capsid of the AAV particle and ABD1 binds to the molecule on the cell surface.
  • Fc1 and Fc2 form an Fc heterodimer.
  • the Fc1 and/or Fc2 in the Fc heterodimer comprise a knob-in- hole mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid substitutions S354C and T366W (according to EU numbering) as compared to a wild type Fc domain
  • the other of Fc1 and Fc2 in the Fc heterodimer comprises amino acid ⁇ ⁇ Attorney Docket No.250298.000954 substitutions Y349C, T366S, L368A, and Y407V (according to EU numbering) as compared to a wild type Fc domain.
  • at least one of Fc1 and Fc2 in the Fc heterodimer comprises a star mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid mutations H435R and/or Y436F (according to EU numbering) as compared to a wild type Fc domain.
  • Fc1 and/or Fc2 comprises the amino acid sequence of SEQ ID NO: 1345 or 1365, or a variant thereof.
  • the first light chain and the second light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof.
  • ABD1 or ABD2 bind to the capsid of the AAV particle, and ABD1 or ABD2 comprises: h) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10; or i) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 71, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • ABD1 or ABD2 binds to the capsid of the AAV particle, and ABD1 or ABD2 comprises: j) an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or k) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 71, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof.
  • ABD1 or ABD2 binds to the capsid of the AAV particle
  • ABD1 or ABD2 comprises: l) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or m) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 73, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 75, and an HCDR3 comprising the ⁇ ⁇ Attorney Docket No.250298.000954 amino acid sequence of SEQ ID NO: 77; and/or an LCDR1 comprising the amino acid sequence of SEQ
  • the molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary subunit gamma 1 (CACNG1). [0020] In some embodiments, the molecule on the cell surface is TfR.
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • ABD1 or ABD2 binds to TfR
  • ABD1 or ABD2 comprises: n) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; o) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 511, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; p) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of
  • ABD1 or ABD2 binds to TfR
  • ABD1 or ABD2 comprises: r) an HCVR that comprises the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; s) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 511, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof; t) an HCVR that comprises the amino acid sequence of SEQ ID NO: 531, or a variant thereof and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or u) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 552, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID ⁇ ⁇ Attorney Docket No.250298.0009
  • ABD1 or ABD2 binds to TfR
  • ABD1 or ABD2 comprises: v) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 502, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 503, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 504; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; w) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 513, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 514; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an
  • a multispecific antibody or multispecific antigen- binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a second heavy chain region of a second Fab (“Fab2”) operably linked to a second Fc domain (“Fc2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); and d) a fourth polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form Fab2, wherein Fab2 comprises a second antigen-binding domain (“ABD2”), wherein e) ABD1 bind
  • the first light chain and the second light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof.
  • ABD1 binds to the capsid of the AAV particle, and ABD1 comprises: j) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or k) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • ABD1 binds to the capsid of the AAV particle, and ABD1 comprises: l) an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or m) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 71, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof.
  • ABD1 binds to the capsid of the AAV particle, and ABD1 comprises: n) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or o) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 73, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 75, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 77; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR
  • ABD2 binds to TfR, and ABD2 comprises: p) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the ⁇ ⁇ Attorney Docket No.250298.000954 amino acid sequence of SEQ ID NO: 501, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; q) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 511, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; r) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the ⁇ ⁇
  • ABD2 binds to TfR, and ABD2 comprises: t) an HCVR that comprises the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; u) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 511, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof; v) an HCVR that comprises the amino acid sequence of SEQ ID NO: 531, or a variant thereof and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or w) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 552, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof.
  • ABD1 or ABD2 binds to TfR
  • ABD1 or ABD2 comprises: x) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 502, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 503, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 504; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; y) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 513, and an HCDR3 comprising the ⁇ ⁇ Attorney Docket No.250298.000954 amino acid sequence of SEQ ID NO: 514; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”), said Fc1 operably linked to a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a third heavy chain region of a third Fab (“Fab3”) operably linked to a second Fc domain (“Fc2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form Fab2, wherein
  • two of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and one of ABD1, ABD2, and ABD3 binds to the molecule on the cell surface.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD3 binds to the molecule on the cell surface.
  • ABD3 binds to the capsid of the AAV particle, and ABD1 and ABD2 bind to the molecule on the cell surface.
  • Fc1 and Fc2 form an Fc heterodimer.
  • the Fc1 and/or Fc2 in the Fc heterodimer comprise a knob-in- hole mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid substitutions S354C and T366W (according to EU numbering) as compared to a wild type Fc domain
  • the other of Fc1 and Fc2 in the Fc heterodimer comprises amino acid substitutions Y349C, T366S, L368A, and Y407V (according to EU numbering) as compared to a wild type Fc domain.
  • At least one of Fc1 and Fc2 in the Fc heterodimer comprises a star mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid mutations H435R and/or Y436F (according to EU numbering) as compared to a wild type Fc domain.
  • Fc1 and/or Fc2 comprises the amino acid sequence of SEQ ID NO: 1345 or 1365, or a variant thereof.
  • the first light chain, the second light chain, and the third light chain comprise the amino acid sequence of SEQ ID NO: 20 or 813, or a variant thereof.
  • the first light chain and the second light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof
  • the third light chain comprises the amino sequence of SEQ ID NO: 813, or a variant thereof.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10.
  • the linker is or comprises a multimer of G4S (SEQ ID NO: 242). [0047] In some embodiments, the linker is G4Sx3 (SEQ ID NO: 1115). [0048] In some embodiments, at least one of ABD1, ABD2, and ABD3 binds to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: i) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid ⁇ ⁇ Attorney Docket No.250298.000954 sequence of SEQ ID NO: 10 or 803, or a variant thereof; or j) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid
  • At least one of ABD1, ABD2, and ABD3 binds to the capsid of the AAV particle
  • ABD1, ABD2, ABD3 comprises: k) an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10 or 803, or a variant thereof; or l) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 71, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10 or 803, or a variant thereof.
  • At least one of ABD1, ABD2, and ABD3 binds to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: m) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12 or 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14 or 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16 or 809; or n) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 73, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 75, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 77; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO:
  • the molecule on the cell surface is ASGR1, TfR, or CACNG1. [0052] In some embodiments, the molecule on the cell surface is TfR. [0053] In some embodiments, the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: o) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; p) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the ⁇ ⁇ Attorney Docket No.250298.000954 amino acid sequence of SEQ ID NO: 511, or a variant thereof
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: s) an HCVR that comprises the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; t) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 511, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof; u) an HCVR that comprises the amino acid sequence of SEQ ID NO: 531, or a variant thereof and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or v) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 552, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO:
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: w) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 502, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 503, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 504; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; x) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 513, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 514; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ
  • the molecule on the cell surface is CACNG1.
  • the other(s) of ABD1, ABD2, and ABD3 binds to CACNG1, and ABD1 or ABD2 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • the other(s) of ABD1, ABD2, and ABD3 binds to CACNG1, and ABD1 or ABD2 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • the other(s) of ABD1, ABD2, and ABD3 binds to CACNG1, and ABD1 or ABD2 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 797, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 799, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 801; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”), said Fc1 operably linked to a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a third heavy ⁇ ⁇ Attorney Docket No.250298.000954 chain region of a third Fab (“Fab3”) operably linked to a second Fc domain (“Fc2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain
  • the first light chain, the second light chain, and the third light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: k) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 803, or a variant thereof; or l) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 803, or a variant thereof.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: m) an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or n) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 71, or a ⁇ ⁇ Attorney Docket No.250298.000954 variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: o) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or p) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 73, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 75, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 77; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO:
  • ABD3 binds to TfR, and ABD3 comprises: q) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; r) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 511, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof; s) an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 531,
  • ABD3 binds to TfR, and ABD3 comprises: u) an HCVR that comprises the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; v) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 511, or a ⁇ ⁇ Attorney Docket No.250298.000954 variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof; w) an HCVR that comprises the amino acid sequence of SEQ ID NO: 531, or a variant thereof and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof; or x) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 552, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 10, or a variant thereof;
  • ABD3 binds to TfR, and ABD3 comprises: y) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 502, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 503, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 504; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; z) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 513, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 514; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”), said Fc1 operably linked to a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a third heavy chain region of a third Fab (“Fab3”) operably linked to a second Fc domain (“Fc2”); ⁇ ⁇ Attorney Docket No.250298.000954 c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain
  • the first light chain and the second light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof.
  • the third light chain comprises the amino acid sequence of SEQ ID NO: 813, or a variant thereof.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ⁇ ⁇ Attorney Docket No.250298.000954 ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.
  • ABD3 binds to CACNG1, and ABD3 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • ABD3 binds to CACNG1, and ABD3 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • ABD3 binds CACNG1, and ABD3 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 797, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 799, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 801; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”), said Fc1 operably linked to a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a third heavy chain region of a third Fab (“Fab3”) operably linked to a second Fc domain (“Fc2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form Fab2, wherein
  • the first light chain, the second light chain, and the third light chain comprise the amino acid sequence of SEQ ID NO: 813, or a variant thereof.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD1 and ABD2 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • ABD3 binds to CACNG1, and ABD3 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • ABD3 binds to CACNG1, and ABD3 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • ABD3 binds CACNG1, and ABD3 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 797, an HCDR2 comprising ⁇ ⁇ Attorney Docket No.250298.000954 the amino acid sequence of SEQ ID NO: 799, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 801; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to an Fc domain, said Fc domain operably linked to a first scFv comprising a first antigen-binding domain (“ABD1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a second heavy chain region of a second Fab (“Fab2”) operably linked to an Fc domain, said Fc domain operably linked to a second scFv comprising a second antigen-binding domain (“ABD2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a third antigen-binding domain (“ABD3”); and d) a
  • two of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD3 and ABD4 binds to CACNG1.
  • ABD3 and ABD4 bind to the capsid of the AAV particle, and ABD1 and ABD2 binds to CACNG1.
  • the Fc domain comprises the amino acid sequence of SEQ ID NO: 1397, or a variant thereof.
  • the first light chain and the second light chain comprise the amino acid sequence of SEQ ID NO: 813, or a variant thereof.
  • the first scFv and/or the second scFv are linked to the Fc domain via a linker.
  • the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10. ⁇ ⁇ Attorney Docket No.250298.000954 [0099]
  • the linker is or comprises a multimer of G4S (SEQ ID NO: 242).
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • at least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 1159, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 1157, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 1159, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 1157, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 797, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 799, and an HCDR3 comprising the amino acid ⁇ ⁇ Attorney Docket No.250298.000954 sequence of SEQ ID NO: 801; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first scFv comprising a first antigen-binding domain (“ABD1”) operably linked to an Fc domain, said Fc domain operably linked to a first heavy chain region of a first Fab (“Fab1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a second scFv comprising a second antigen-binding domain (“ABD2”) operably linked to an Fc domain, said Fc domain operably linked to a second heavy chain region of a second Fab (“Fab2”) ; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a third antigen-binding domain (“ABD3”); and d)
  • two of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1.
  • ABD1 and ABD2 bind to the capsid of the AAV particle, and ABD3 and ABD4 binds to CACNG1.
  • ABD3 and ABD4 bind to the capsid of the AAV particle, and ABD1 and ABD2 binds to CACNG1.
  • the Fc domain comprises the amino acid sequence of SEQ ID NO: 1397, or a variant thereof.
  • the first light chain and the second light chain comprise the sequence of SEQ ID NO: 20 or 813, or a variant thereof.
  • the first Fab and/or the second Fab is linked to the Fc domain via a linker.
  • the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10.
  • the linker is or comprises a multimer of G4S (SEQ ID NO: ⁇ ⁇ Attorney Docket No.250298.000954 242).
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • at least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10 or 1157, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803 or 1352, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 797, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 799, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 801; and/or an LCDR1 comprising the amino acid sequence of ⁇ ⁇ Attorney Docket No.250298.000954 SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to an Fc domain, said Fc domain operably linked to a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a third heavy chain region of a third Fab (“Fab3”) operable liked to an Fc domain, said Fc domain operably linked to a fourth heavy chain region of a fourth Fab (“Fab4”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second
  • two of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1.
  • ABD1 and ABD3 bind to the capsid of the AAV particle, and ABD2 and ABD4 binds to CACNG1.
  • ABD2 and ABD4 bind to the capsid of the AAV particle, and ABD1 and ABD3 binds to CACNG1.
  • the first light chain, the second light chain, the third light chain, and the fourth light chain comprise the amino acid sequence of SEQ ID NO: 813, or a variant thereof.
  • the Fc domain is linked to the second heavy chain region ⁇ ⁇ Attorney Docket No.250298.000954 and the fourth heavy chain region via a linker.
  • the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10.
  • the linker is or comprises a multimer of G4S (SEQ ID NO: 242).
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • at least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 1351, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 803, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to ⁇ ⁇ Attorney Docket No.250298.000954 CACNG1, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 797, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 799, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 801; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 805, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 807, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 809.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to an Fc domain, said Fc domain operably linked to a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a third heavy chain region of a third Fab (“Fab3”) operable liked to an Fc domain, said Fc domain operably linked to a fourth heavy chain region of a fourth Fab (“Fab4”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second
  • two of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to TfR.
  • ABD1 and ABD3 bind to TfR, and ABD2 and ABD4 binds to the capsid of the AAV particle.
  • ABD2 and ABD4 bind to TfR, and ABD1 and ABD3 binds to the capsid of the AAV particle.
  • the first light chain, the second light chain, the third light chain, and the fourth light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof.
  • the Fc domain is linked to the second heavy chain region and the fourth heavy chain region via a linker.
  • the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10.
  • the linker is or comprises a multimer of G4S (SEQ ID NO: 242).
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • at least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • At least one of ABD1, ABD2, ABD3, and ABD4 bind to the capsid of the AAV particle, and ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to TfR
  • ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to TfR, and ABD1, ABD2, ABD3, and/or ABD4 comprises: ⁇ ⁇ Attorney Docket No.250298.000954 an HCVR that comprises the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 binds to TfR
  • ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 502, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 503, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 504; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”), said Fc1 operably linked to a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a second Fc domain (“Fc2”), said Fc2 operably linked to a third heavy chain region of a third Fab (“Fab3”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form
  • two of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and one of ABD1, ABD2, and ABD3 binds to TfR.
  • two of ABD1, ABD2, and ABD3 bind to TfR, and one of ABD1, ABD2, and ABD3 binds to the capsid of the AAV particle.
  • ABD1 binds to TfR, and ABD2 and ABD3 binds to the ⁇ ⁇ Attorney Docket No.250298.000954 capsid of the AAV particle.
  • ABD1 binds to the capsid of the AAV particle, and ABD2 and ABD3 binds to TfR.
  • Fc1 and Fc2 form an Fc heterodimer.
  • the Fc1 and/or Fc2 in the Fc heterodimer comprise a knob- in-hole mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid substitutions S354C and T366W (according to EU numbering) as compared to a wild type Fc domain
  • the other of Fc1 and Fc2 in the Fc heterodimer comprises amino acid substitutions Y349C, T366S, L368A, and Y407V (according to EU numbering) as compared to a wild type Fc domain.
  • at least one of Fc1 and Fc2 in the Fc heterodimer comprises a star mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid mutations H435R and/or Y436F (according to EU numbering) as compared to a wild type Fc domain.
  • Fc1 and/or Fc2 comprises the amino acid sequence of SEQ ID NO: 1397 or 1511, or a variant thereof.
  • the first light chain, the second light chain, and the third light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof.
  • Fc1 and/or Fc2 is linked to the second heavy chain region and the third heavy chain region via a linker.
  • the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10. [00167] In some embodiments, the linker is or comprises a multimer of G4S (SEQ ID NO: 242). [00168] In some embodiments, the linker is G4Sx3 (SEQ ID NO: 1115).
  • At least one of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • At least one of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, ⁇ ⁇ Attorney Docket No.250298.000954 and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • At least one of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR
  • ABD1, ABD2, ABD3, and/or ABD4 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 502, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 503, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 504; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 13, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 14.
  • ABD1 binds to TfR; j) ABD2 and ABD3 bind to the capsid of the AAV particle; k) Fc1 comprises the amino acid sequence of SEQ ID NO: 1397, or a variant thereof; and l) Fc2 comprises the amino acid sequence of SEQ ID NO: 1511, or a variant thereof.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation: a first heavy chain region of a first Fab (“Fab1”) operably linked to a first Fc domain (“Fc1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation: a second heavy ⁇ ⁇ Attorney Docket No.250298.000954 chain region of a second Fab (“Fab2”) operably linked to a second Fc domain (“Fc2”), said Fc2 operably linked to a third heavy chain region of a third Fab (“Fab3”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain
  • two of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and one of ABD1, ABD2, and ABD3 binds to TfR.
  • two of ABD1, ABD2, and ABD3 bind to TfR, and one of ABD1, ABD2, and ABD3 binds to the capsid of the AAV particle.
  • ABD2 and ABD3 bind to the capsid of the AAV particle, and ABD1 binds to TfR.
  • ABD2 and ABD3 bind to TfR, and ABD1 binds to the capsid of the AAV particle.
  • Fc1 and Fc2 form an Fc heterodimer.
  • the Fc1 and/or Fc2 in the Fc heterodimer comprise a knob- in-hole mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid substitutions S354C and T366W (according to EU numbering) as compared to a wild type Fc domain
  • the other of Fc1 and Fc2 in the Fc heterodimer comprises amino acid substitutions Y349C, T366S, L368A, and Y407V (according to EU numbering) as compared to a wild type Fc domain.
  • at least one of Fc1 and Fc2 in the Fc heterodimer comprises a star mutation as compared to a wild type Fc domain.
  • one of Fc1 and Fc2 in the Fc heterodimer comprises amino acid mutations H435R and/or Y436F (according to EU numbering) as compared to a wild type Fc domain.
  • Fc1 and/or Fc2 comprises the amino acid sequence of SEQ ID NO: 1397 or 1511, or a variant thereof.
  • the first light chain, the second light chain, and the third light chain comprise the amino acid sequence of SEQ ID NO: 20, or a variant thereof.
  • Fc2 is linked to the third heavy chain region via a linker.
  • the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10.
  • the linker is or comprises a multimer of G4S (SEQ ID NO: 242).
  • the linker is G4Sx3 (SEQ ID NO: 1115).
  • At least one of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • At least one of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO: 52, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • At least one of ABD1, ABD2, and ABD3 bind to the capsid of the AAV particle, and ABD1, ABD2, and/or ABD3 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: an HCVR that comprises an HCDR1, HCDR2, and HCDR3 of an HCVR comprising the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and/or an LCVR that comprises an LCDR1, LCDR2, and LCDR3 of an LCVR comprising the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: ⁇ ⁇ Attorney Docket No.250298.000954 an HCVR that comprises the amino acid sequence of SEQ ID NO: 501, or a variant thereof, and an LCVR that comprises the amino acid sequence of SEQ ID NO: 10, or a variant thereof.
  • the other(s) of ABD1, ABD2, and ABD3 binds to TfR, and ABD1, ABD2, and/or ABD3 comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO: 502, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 503, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 504; and/or an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 13, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 14.
  • ABD1 binds to TfR; j) ABD2 and ABD3 bind to the capsid of the AAV particle; k) Fc1 comprises the amino acid sequence of SEQ ID NO: 1397, or a variant thereof; and l) Fc2 comprises the amino acid sequence of SEQ ID NO: 1511, or a variant thereof.
  • the AAV is wild type.
  • the AAV particle comprises one or more mutations in one or more AAV capsid proteins inhibiting the natural tropism of said AAV particle.
  • the multispecific antibody or multispecific antigen-binding fragment is a bispecific antibody or bispecific antigen-binding fragment thereof.
  • a pharmaceutical composition comprising the multispecific antibodies or multispecific antigen-binding fragments as described herein, and a pharmaceutically acceptable carrier or excipient.
  • a molecular complex comprising an AAV particle bound to one or more multispecific antibodies and/or multispecific antigen-binding fragments as described herein.
  • the AAV particle comprises one or more mutations in one or more AAV capsid proteins inhibiting the natural tropism of said AAV particle.
  • a pharmaceutical composition comprising the molecular complex as described herein and a pharmaceutically acceptable carrier or excipient.
  • a method of preparing the molecular complex as described herein comprising incubating the AAV particle in the presence of the one or more multispecific antibodies and/or multispecific antigen-binding fragments under conditions allowing specific binding of said one or more multispecific antibodies and/or multispecific antigen-binding fragments to said AAV particle capsid.
  • a method for targeting an AAV particle to a ⁇ ⁇ Attorney Docket No.250298.000954 cell expressing a molecule on the cell surface comprising contacting the cell with the molecular complex as described herein, or the pharmaceutical composition as described herein, wherein said molecular complex comprises one or more multispecific antibodies and/or multispecific antigen-binding fragments which bind to said molecule on the cell surface.
  • a method for delivering a polynucleotide to a cell expressing a molecule on the cell surface comprising contacting the cell with the molecular complex as described herein or the pharmaceutical composition as described herein, wherein said molecular complex comprises the AAV particle comprising said polynucleotide and bound to one or more multispecific antibodies and/or multispecific antigen-binding fragments which bind to said molecule on the cell surface.
  • the cell is in a subject and the molecular complex is administered to the subject.
  • the AAV particle does not target the cell in the absence of the one or more multispecific antibodies and/or multispecific antigen-binding fragments.
  • any of the features or components of embodiments discussed above or herein may be combined, and such combinations are encompassed within the scope of the present disclosure. Any specific value discussed above or herein may be combined with another related value discussed above or herein to recite a range with the values representing the upper and lower ends of the range, and such ranges are encompassed within the scope of the present disclosure.
  • Other embodiments will become apparent from a review of the ensuing detailed description.
  • Figures 1A-1L depict graphs of binding activities of the anti-adeno-associated virus (AAV) antibodies described herein to various adeno-associated virus (AAV) serotypes. RLU, relative light units.
  • Figures 2A-2J depict graphs showing immunoglobulin gamma Fc receptor I (FcGR- 1) AAV retargeting by anti-AAV antibodies derived using next-generation sequencing (NGS)/mass spectrometry (MS) methods ( Figures 2A-2F) and by anti-AAV antibodies derived from hybridoma ( Figures 2G-2J) described herein for various AAV serotypes.
  • FcGR- 1 depict graphs showing immunoglobulin gamma Fc receptor I (FcGR- 1) AAV retargeting by anti-AAV antibodies derived using next-generation sequencing (NGS)/mass spectrometry (MS) methods ( Figures 2A-2F) and by anti-AAV antibodies derived from hybridoma ( Figures 2G-2J)
  • FIG 3 shows a schematic representation of alternative format antibody designs for bispecific antibodies of the present disclosure.
  • Exemplary bispecific antibodies target AAV and Asialoglycoprotein Receptor 1 (ASGR1).
  • Star mutations are depicted with an asterisk (*) and knob-in-hole (KiH) mutations are depicted with a triangle (e.g., ⁇ ).
  • ⁇ Attorney Docket No.250298.000954
  • Figure 4 displays a description of alternative format bispecific antibody molecules described herein.
  • Figure 5 illustrates an example rationale for anti-AAV antibody selection.
  • Figures 6A-6J depict graphs of binding affinities of alternative format antibodies described herein (e.g., anti-AAV x anti-ASGR1 [also called AAV x ASGR1, and the like, herein] bispecific antibodies) to an AAV2 heparin binding mutant (HBM) serotype.
  • the AAV2-HBM serotype contains R585A, R588A mutations. RLU, relative light units.
  • Figures 7A-7B illustrate that bispecific antibodies can effectively retarget adeno- associated virus (AAV) to the central nervous system (CNS).
  • AAV adeno- associated virus
  • FIG. 7A A schematic representation of anti-transferrin receptor (TfR) x anti-AAV bispecific antibody retargeting of AAV to the CNS is depicted in Figure 7A.
  • BBB blood-brain barrier.
  • Figures 8A-8B demonstrate that liver detargeting can be controlled by modulating antibody concentration [Ab] and/or use of a detargeting capsid.
  • a comparison of transduction efficiency between the covalent platform and the bispecific platform evaluated using cDNA quantitation is shown in Figure 8A.
  • Figure 9 shows a schematic representation of anti-AAV x anti-CACNG1 (also called AAV x CACNG1, and the like, herein) alternative format (AF) antibodies AF70 and AF71.
  • AF70 comprises anti-AAV#70 scFv fused to the N-terminus of anti-CACNG1 REGN10717 hIgG1 N297G antibody.
  • AF71 comprises anti-AAV#70 scFv fused to the C-terminus of anti-CACNG1 REGN10717 hIgG1 N297G antibody.
  • FIG. 10 shows flow cytometry data illustrating co-incubation of AAV9 W503A with AF70 (top two rows) and AF71 (bottom two rows) resulted in improved transduction efficiency over control AAV without antibody on HEK 293 cells overexpressing mouse CACNG1 (mCACNG1). Transduction efficiency was optimal between molar ratios of 1 AAV to 1 antibody (AAV:Ab ratio 1:1) and 1 AAV to 9 antibodies (AAV:Ab ratio 1:9) as assessed by percent (%) GFP positive cells.
  • FIG 11 depicts line graphs showing co-incubation of AAV9 W503A with AF70 (top) and AF71 (bottom) resulted in improved transduction efficiency over control AAV without antibody on HEK 293 cells overexpressing mouse CACNG1 (mCACNG1). Transduction efficiency was optimal between molar ratios of 1 AAV to 1 antibody (AAV:Ab ratio 1:1) and 1 AAV to 9 antibodies (AAV:Ab ratio 1:9) as assessed by mean fluorescence intensity (MFI).
  • MFI mean fluorescence intensity
  • Figure 12 shows flow cytometry data illustrating co-incubation of AAV9 W503A with ⁇ ⁇ Attorney Docket No.250298.000954 AF70 (top two rows) and AF71 (bottom two rows) resulted in improved AAV transduction efficiency over control AAV without antibody on HEK 293 cells overexpressing human CACNG1 (hCACNG1). Transduction efficiency was optimal between molar ratios of 1 AAV to 1 antibody (AAV:Ab ratio 1:1) and 1 AAV to 9 antibodies (AAV:Ab ratio 1:9) as assessed by percent (%) GFP positive cells.
  • FIG. 13 depicts line graphs showing co-incubation of AAV9 W503A with AF70 (top) and AF71 (bottom) resulted in improved AAV transduction efficiency over control AAV without antibody on HEK 293 cells overexpressing human CACNG1 (hCACNG1). Transduction efficiency was optimal between molar ratios of 1 AAV to 1 antibody (AAV:Ab ratio 1:1) and 1 AAV to 9 antibodies (AAV:Ab ratio 1:9) as assessed by mean fluorescence intensity (MFI).
  • MFI mean fluorescence intensity
  • Figure 14 shows immunohistochemical staining of differentiated C2C12 myotubes for Myosin Heavy Chain (MyHC) and demonstration that incubation of AAV with AF70 and AF71 enhanced transduction into the myotubes as determined by GFP fluorescence.
  • Figure 15 shows immunohistochemical staining of differentiated human myotubes for Myosin Heavy Chain (MyHC) and demonstration that incubation of AAV with AF70 and AF71 enhanced transduction into the myotubes as determined by GFP fluorescence. Human myotubes were most efficiently transduced with AAV complexed with AF71 at molar ratio 1:3 and 1:9.
  • Figures 16A-16D demonstrate incubation of AAV with AF70 and AF71 enhanced transduction into differentiated C2C12 myotubes as determined by quantification of GFP positive cells.
  • Figures 17A-17D demonstrate incubation of AAV with AF70 and AF71 enhanced transduction into differentiated human myotubes as determined by quantification of GFP positive cells.
  • Figures 18A-18B show non-limiting examples of anti-AAV x anti-mTfR (also called AAV x mTfR, and the like, herein) alternative format (AF) antibodies described herein.
  • anti-AAV x anti-mTfR also called AAV x mTfR, and the like, herein
  • AF alternative format
  • FIG. 19 shows an example experimental setup used to test alternative format antibodies binding to AAV9W503A virus by ELISA.
  • Figure 20 depicts ELISA data for anti-AAV x anti-mTfR alternative format antibodies binding to AAV9W503A.
  • Figure 21 shows a schematic diagram of a FLuc assay protocol used to test retargeting of AAV9W503A using mTfR alternative format antibodies on 293T cells expressing mTfR receptor.
  • Figure 22 shows a line graph of data generated in experiments testing AAV9W503A retargeting using anti-AAV x anti-mTfR alternative format antibodies on mTfR293T cells.
  • Figures 23A-23C illustrate a retargeting assay using AAV9 scCBH.eGFP.
  • Figures 24A-24B show in vitro test infection results for in vivo injection samples.
  • Figures 25A-25F depict anti-AAV x anti-mTfR liver and brain (hippocampus, cortex, and cerebellum) green fluorescent protein (GFP) staining for control groups ( Figure 25A) and alternative format designs, anti-AAV x anti-mTfR AF1 ( Figure 25B), anti-AAV x anti-mTfR AF3 ( Figure 25C), anti-AAV x anti-mTfR AF5 ( Figure 25D), anti-AAV x anti-mTfR AF7 ( Figure 25E), and anti-AAV x anti-mTfR AF9 (Figure 25F).
  • Figure 26 shows relative RNA expression of GFP in liver samples as determined by RT-qPCR.
  • Figure 27 shows RNA expression of GFP in brain samples as determined by RT- qPCR.
  • Figures 28A-28D depict GFP staining in brain, heart, and liver tissues from mice receiving AAV9 ( Figures 28A-28B) or AAV9W503A ( Figures 28C and 28D) complexed with anti-AAV x anti-mTfR alternative format AF7 at AAV vector genome (VG) to antibody (Ab) ratios (VG:Ab ratios) of 1:9 and 1:3.
  • Figures 29A-29I depict anti-AAV x anti-mTfR alternative format designs AF3 and AF7.
  • Figure 29A provides a description of anti-AAV x anti-mTfR alternative format designs AF3 and AF7. Star mutations are depicted with an asterisk (*) and knob-in-hole (KiH) mutations are depicted with a triangle (e.g., ⁇ ).
  • Figures 29B-29E show examples of anti-AAV x anti-mTfR AF3 amino acid sequences ( Figure 29B) and corresponding nucleotide sequences ( Figures 29C-29E).
  • Figures 29F-29I show examples of anti-AAV x anti-mTfR AF7 amino acid sequences ( Figure 29F) and corresponding nucleotide sequences ( Figures 29G- 29I).
  • Figures 30A-30G depict anti-AAV x anti-CACNG1 alternative format designs AF70 and AF71.
  • Figure 30A provides a description of anti-AAV x anti-CACNG1 alternative format designs AF70 and AF71.
  • Figures 30B-30G show examples of anti-AAV x anti-CACNG1 AF70 amino acid sequences ( Figure 30B) and corresponding nucleotide sequences ( Figures 30C- 30D).
  • Figures 30E-30G show examples of anti-AAV x anti-CACNG1 AF71 amino acid sequences ( Figure 30E) and corresponding nucleotide sequences ( Figures 30F-30G).
  • Figure 31 illustrates anti-AAV x anti-CACNG1 bispecific antibody enhancement of transduction into CACNG1 overexpressing 293 cells. GFP expression was assessed by flow cytometry and is shown as % GFP positive cells (top panels) or MFI of GFP expressing cells (bottom panels). ⁇ ⁇ Attorney Docket No.250298.000954 [00244]
  • Figure 32 depicts an example of a study design for testing Hu37 complexed with anti-AAV x anti-CACNG12x2 antibodies in D2.MDX mice.
  • Figure 33 shows in vitro transduction in HEK293-hCACNG1 of complexes prepared for in vivo study. ⁇
  • Figures 34A-34C illustrate improved Hu37 transduction in muscle tissues when complexed with an 2x2 anti-AAV x anti-CACNG1 alternative format antibody.
  • Figures 35A-35B illustrate bispecific antibody enhancement of Hu37 transduction into skeletal muscle.
  • Figures 36A-36B depict anti-AAV x anti-ASGR alternative format designs AF21 (REGN16199), AF22 (REGN16200), AF29, AF30 (REGN16204), AF32 (REGN16202), AF37, AF41, AF60A, AF61A, AF62, and AF63.
  • Figure 36A provides a description of anti-AAV x anti- ASGR alternative format designs AF21 (REGN16199), AF22 (REGN16200), AF29, AF30 (REGN16204), and AF32 (REGN16202).
  • Figure 36B provides a description of anti-AAV x anti-ASGR alternative format designs AF37, AF41, AF60A, AF61A, AF62, and AF63.
  • Figure discloses “3xG4S” as SEQ ID NO: 1115 and “4xG4S” as SEQ ID NO: 1161.
  • Figures 37A-37B depict IVIS (In Vivo Imaging Instrument, Perkin Elmer) data illustrating retargeting of AAV to liver with AF30 and AF32.
  • Figure 38 depicts IVIS data that demonstrate retargeting of multiple serotypes to liver with AF30.
  • Figures 39A-39D depict IVIS & PK qPCR data that demonstrate retargeting to liver with antibodies comprised of either one or two anti-AAV binding arms.
  • Figures 40A-40C depict IVIS, PK qPCR, & ELISA data that demonstrate antibody can effectively retarget AAV when dosed prior to AAV administration.
  • Figures 41A-41B depict IVIS & PK qPCR data that demonstrate retargeting of REGN16199 with and without Fc.
  • Figures 42A-42F depict IVIS, PK qPCR, & RNA Taqman data that demonstrate successful targeting with linear Fab formats.
  • Figure 43 depicts PK qPCR data of serum samples 24 hours after administration of various antibody formats complexed with AAV.
  • Figure 44 depicts IVIS data that demonstrate retargeting of AAV with a variety of different antibody formats.
  • Figure 45 depicts IVIS data that demonstrate retargeting of AAV with antibody comprised of one anti-AAV arm and one anti-ASGR1 arm.
  • Figures 46A-46I depict anti-AAV x anti-ASGR alternative format designs AF1, AF5, and AF9.
  • Figures 46A-46C show examples of anti-AAV x anti-ASGR AF1 amino acid sequences ( Figure 46A) and corresponding nucleotide sequences ( Figures 46B-46C).
  • Figures 46D-46F show examples of anti-AAV x anti-ASGR AF5 amino acid sequences ( Figure 46D) and corresponding nucleotide sequences ( Figures 46E-46F).
  • Figures 46G-46I show examples of anti-AAV x anti-ASGR AF9 amino acid sequences ( Figure 46G) and corresponding nucleotide sequences ( Figures 46H-46I).
  • Figures 47A-47I depict anti-AAV x anti-mTfR alternative format designs AF1, AF5, and AF9.
  • Figures 47A-47C show examples of anti-AAV x anti-mTfR AF1 amino acid sequences ( Figure 47A) and corresponding nucleotide sequences ( Figures 47B-47C).
  • Figures 47D-47F show examples of anti-AAV x anti-mTfR AF5 amino acid sequences ( Figure 47D) and corresponding nucleotide sequences ( Figures 47E-47F).
  • Figures 47G-47I show examples of anti-AAV x anti-mTfR AF9 amino acid sequences ( Figure 47G) and corresponding nucleotide sequences ( Figures 47H-47I).
  • Figure discloses “3xG4S” as SEQ ID NO: 1115 and “4xG4S” as SEQ ID NO: 1161.
  • Figure 48 ⁇ shows a schematic representation of anti-AAV x anti-CACNG1 alternative format (AF) antibodies AF70, AF71, AF71x, AF72a, AF72b, AF73, AF74A, AF74B, and AF76.
  • AF anti-AAV x anti-CACNG1 alternative format
  • Figures 49A-49B show flow cytometry data illustrating that co-incubation of AAV Hu37 with AF71, AF71x, AF72a, AF73, AF74A, and AF74B resulted in improved transduction efficiency over control AAV without antibody on HEK 293 cells overexpressing human CACNG1 (hCACNG1).
  • FIGS 50A-50E ⁇ illustrate bispecific antibody enhancement of Hu37 transduction into skeletal muscle. Transduction levels are shown for the following skeletal muscles: tongue (Figure 50A), gastrocnemius (gastroc)/soleus ( Figure 50B), and tibialis anterior (TA) ( Figure 50C). Transduction levels for the heart (Figure 50D) and liver ( Figure 50E) are also depicted.
  • Figures 51A-51C depict examples of amino acid sequences ( Figure 51A) and corresponding nucleotide sequences ( Figures 51B-51C) for anti-AAV x anti-CACNG1 alternative format design AF71x.
  • Figures 52A-52C depict examples of amino acid sequences ( Figure 52A) and corresponding nucleotide sequences ( Figures 52B-52C) for anti-AAV x anti-CACNG1 alternative format design AF72a.
  • Figures 53A-53C depict examples of amino acid sequences ( Figure 53A) and ⁇ ⁇ Attorney Docket No.250298.000954 corresponding nucleotide sequences ( Figures 53B-53C) for anti-AAV x anti-CACNG1 alternative format design AF72b.
  • Figures 54A-54C depict examples of amino acid sequences ( Figure 54A) and corresponding nucleotide sequences ( Figures 54B-54C) for anti-AAV x anti-CACNG1 alternative format design AF73.
  • Figures 55A-55E depict examples of amino acid sequences ( Figures 55A-55B) and corresponding nucleotide sequences ( Figures 55C-55E) for anti-AAV x anti-CACNG1 alternative format design AF74A.
  • Figures 56A-56D depict examples of amino acid sequences ( Figure 56A) and corresponding nucleotide sequences ( Figures 56B-56D) for anti-AAV x anti-CACNG1 alternative format design AF74B.
  • Figures 57A-57C depict examples of amino acid sequences ( Figure 57A) and corresponding nucleotide sequences ( Figures 57B-57C) for anti-AAV x anti-CACNG1 alternative format design AF76.
  • Figures 58A-58B ⁇ show schematic representations of anti-AAV x anti-human transferrin receptor (hTfR) (also called AAV x hTfR herein) antibodies DB7035B, DB7043B, DB7045B, DB7047B, DB8035B, DB8043B, DB8045B, and DB8047B (Davis body orientation), as well as alternative format (AF) (i.e., Altibody) antibodies AF7035B, AF7043B, AF7045B, AF7047B, AF8035B, AF8043B, AF8045B, and AF8047B.
  • hTfR anti-AAV x anti-human transferrin receptor
  • Figure 58B discloses “3xG4S” as SEQ ID NO: 1115.
  • Figures 59A-59B show flow cytometry data illustrating co-incubation of AAV9 with DB7035B, DB7043B, DB7045B, DB7047B, DB8035B, DB8043B, DB8045B, and DB8047B, resulted in improved transduction efficiency over control AAV9 without antibody on 3T3 cells overexpressing human TfR (hTfR).
  • FIGS. 60A-60B show flow cytometry data illustrating co-incubation of AAV9 with AF7035B, AF7043B, AF7045B, AF7047B, AF8035B, AF8043B, AF8045B, and AF8047B, resulted in improved transduction efficiency over control AAV9 without antibody on 3T3 cells overexpressing human TfR (hTfR). Transduction efficiency was assessed by percent (%) GFP positive cells ( Figure 60A) and Median Fluorescence Intensity (MFI) ( Figure 60B).
  • Figures 61A-61D illustrate bispecific antibody enhancement of AAV9 transduction into different brain areas. Bispecific antibody enhancement of AAV9 transduction into the hippocampus, cerebellum, and cortex are shown in Figures 61A-61C, respectively. ⁇ In addition to hippocampus (CA2 region), cerebellum, and cortex, Figure 61D illustrates bispecific antibody AF7045B enhancement of AAV9 transduction into medulla, olfactory bulb, thalamus, ⁇ ⁇ Attorney Docket No.250298.000954 striatum, hypothalamus, and midbrain.
  • Figure 62 illustrates quantification of green fluorescent protein (GFP) in the cortex of hTfR mice (7229KO) by measuring the percent (%) GFP area.
  • Figures 63A-63C illustrate quantification of mRNA levels in the brain ( Figure 63A), liver ( Figure 63B), and heart ( Figure 63C) of hTfR mice (7229KO).
  • Figures 64A-64D depict examples of amino acid sequences ( Figure 64A) and corresponding nucleotide sequences ( Figures 64B-64D) for anti-AAV x anti-hTFR Davis body design DB7035B.
  • Figures 65A-65D depict examples of amino acid sequences ( Figure 65A) and corresponding nucleotide sequences ( Figures 65B-65D) for anti-AAV x anti-hTFR Davis body design DB7043B.
  • Figures 66A-66D depict examples of amino acid sequences ( Figure 66A) and corresponding nucleotide sequences ( Figures 66B-66D) for anti-AAV x anti-hTFR Davis body design DB7045B.
  • Figures 67A-67D depict examples of amino acid sequences ( Figure 67A) and corresponding nucleotide sequences ( Figures 67B-67D) for anti-AAV x anti-hTFR Davis body design DB7047B.
  • Figures 68A-68D depict examples of amino acid sequences ( Figure 68A) and corresponding nucleotide sequences ( Figures 68B-68D) for anti-AAV x anti-hTFR Davis body design DB8035B.
  • Figures 69A-69D depict examples of amino acid sequences ( Figure 69A) and corresponding nucleotide sequences ( Figures 69B-69D) for anti-AAV x anti-hTFR Davis body design DB8043B.
  • Figures 70A-70D depict examples of amino acid sequences ( Figure 70A) and corresponding nucleotide sequences ( Figures 70B-70D) for anti-AAV x anti-hTFR Davis body design DB8045B.
  • Figures 71A-71D depict examples of amino acid sequences ( Figure 71A) and corresponding nucleotide sequences ( Figures 71B-71D) for anti-AAV x anti-hTFR Davis body design DB8047B.
  • Figures 72A-72D depict examples of amino acid sequences ( Figure 72A) and corresponding nucleotide sequences ( Figures 72B-72D) for anti-AAV x anti-hTFR alternative format design AF7035B.
  • Figures 73A-73D depict examples of amino acid sequences ( Figure 73A) and corresponding nucleotide sequences ( Figures 73B-73D) for anti-AAV x anti-hTFR alternative format design AF7043B.
  • Figures 74A-74D depict examples of amino acid sequences ( Figure 74A) and ⁇ ⁇ Attorney Docket No.250298.000954 corresponding nucleotide sequences ( Figures 74B-74D) for anti-AAV x anti-hTFR alternative format design AF7045B.
  • Figures 75A-75D depict examples of amino acid sequences ( Figure 75A) and corresponding nucleotide sequences ( Figures 75B-75D) for anti-AAV x anti-hTFR alternative format design AF7047B.
  • Figures 76A-76D depict examples of amino acid sequences ( Figure 76A) and corresponding nucleotide sequences ( Figures 76B-76D) for anti-AAV x anti-hTFR alternative format design AF8035B.
  • Figures 77A-77D depict examples of amino acid sequences ( Figure 77A) and corresponding nucleotide sequences ( Figures 77B-77D) for anti-AAV x anti-hTFR alternative format design AF8043B.
  • Figures 78A-78D depict examples of amino acid sequences ( Figure 78A) and corresponding nucleotide sequences ( Figures 78B-78D) for anti-AAV x anti-hTFR alternative format design AF8045B.
  • Figures 79A-79D depict examples of amino acid sequences ( Figure 79A) and corresponding nucleotide sequences ( Figures 79B-79D) for anti-AAV x anti-hTFR alternative format design AF8047B.
  • MBMs multispecific binding molecules
  • Figure 80A shows the structure of a MBM comprising: i) a first polypeptide chain comprising (in an N- to C-terminal orientation) a ⁇ single domain antibody (sdAb) (e.g., VHH), an optional linker, a CH2 domain, a CH3 domain, an optional linker, a variable heavy (VH) domain, and a CH1 domain; ii) a second polypeptide chain comprising (in an N- to C-terminal orientation) a variable light (VL) domain and a constant domain of a light chain (CL) associated with the VH and CH1 domains; iii) a third polypeptide chain comprising (in an N- to C-terminal orientation) a ⁇ single-chain Fv (scFv), an optional linker, a CH2 domain, a CH3 domain, an optional linker, a VH domain, and a CH1 domain; and iv) a fourth polypeptide chain comprising (in an N- to
  • Figure 80B shows the structure of a MBM comprising: ⁇ i) a first polypeptide chain comprising (in an N- to C-terminal orientation) a sdAb (e.g., VHH), an optional linker, a CH2 domain, a CH3 domain, an optional linker, a VH domain, and a CH1 domain; ii) a second polypeptide chain comprising (in an N- to C-terminal orientation) a VL domain and a CL domain associated with the VH and CH1 domains; iii) a third polypeptide chain comprising (in an N- to C-terminal orientation) a sdAb (e.g., VHH), an optional linker, a CH2 domain, a CH3 domain, an optional linker, a VH domain, and a CH1 domain; and iv) a fourth polypeptide chain comprising (in an N- to C-terminal orientation) a VL domain and a CL domain associated with the VH and CH1
  • Figures 81A-81B ⁇ show examples of structures of ⁇ anti-AAV x anti-TfR alternative format designs REGN23091 (also referred to as AF7 herein) ( Figure 81A) and REGN22198 ( Figure 81B).
  • Figures 82A-82B ⁇ show examples of structures of ⁇ anti-AAV x anti-CACNG1 alternative format designs REGN20586 (also referred to as AF71 herein) ( Figure 82A) and REGN22008 (also referred to as AF71x herein) ( Figure 82B).
  • Figure 83 illustrates bispecific antibody enhancement of AAV9 transduction into the brain using different AAV and antibody complexing conditions along with sequential dosing of antibody and AAV. F/T, freeze and thaw.
  • Figure 84 illustrates anti-AAV x anti-CACNG1 bispecific antibody enhancement of AAV9 W503A transduction into skeletal muscle. Transduction levels are shown for the following skeletal muscles: gastrocnemius and quadriceps. Transduction level for the heart and liver are also depicted.
  • Figure 85 shows improved transduction efficiency of AAV9 with AFT5 and AFT8 alternative format antibodies over control AAV9 without antibody on 3T3 cells overexpressing human TfR (hTfR).
  • Figures 86A-86B show schematics of AFT5 ( Figure 86A) and AFT8 ( Figure 86B).
  • Figures 87A-87D depict examples of amino acid sequences ( Figure 87A) and corresponding nucleotide sequences ( Figures 87B-87D) for anti-AAV x anti-hTFR alternative format design AFT8.
  • Figures 88A-88C depict examples of amino acid sequences ( Figure 88A) and corresponding nucleotide sequences ( Figures 88B-88C) for anti-AAV x anti-hTFR alternative format design AFT5.
  • Figures 89A-89D depict examples of amino acid sequences ( Figure 89A) and corresponding nucleotide sequences ( Figures 89B-89D) for anti-AAV x anti-hTFR REGN22198.
  • DETAILED DESCRIPTION [00302] Before the present disclosure is described, it is to be understood that this disclosure is not limited to particular methods and experimental conditions described, as such methods and conditions may vary.
  • the term “about”, when used in reference to a particular recited numerical value, means that the value may vary from the recited value by no more than 1%.
  • the expression “about 100” includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).
  • the term “antigen” encompasses any agent (e.g., protein, peptide, polysaccharide, glycoprotein, glycolipid, nucleotide, portions thereof, or combinations thereof) that, when introduced into an immunocompetent host is recognized by the immune system of the host and is capable of eliciting an immune response by the host.
  • epitope can refer to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule known as a paratope.
  • a single antigen may have more than one epitope.
  • different antibodies may bind to different areas on an antigen and may have different biological effects.
  • Epitopes may be either conformational or linear.
  • a conformational epitope is produced by spatially juxtaposed amino acids from different segments of the linear polypeptide chain.
  • a linear epitope is one produced by adjacent amino acid residues in a polypeptide chain.
  • an epitope may include moieties of saccharides, phosphoryl groups, or sulfonyl groups on the antigen.
  • Epitopes may also be defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and are defined as those residues that directly contribute to the affinity of the interaction between a major histocompatibility complex (MHC) molecule and the antigen.
  • MHC major histocompatibility complex
  • antigen-binding molecule refers in its broadest sense to a molecule that specifically binds to an antigen.
  • an antigen-binding molecule is an antibody or an antigen-binding fragment of an antibody, including, e.g., multispecific antibodies such as bispecific antibodies or fragments thereof.
  • multispecific antigen-binding molecule or “multispecific binding molecule” or “MBM” includes molecules (e.g., antibodies and antigen-binding fragments of antibodies) that bind two or more (e.g., three or four) different epitopes or antigens.
  • the multispecific antigen-binding molecules are bispecific (e.g., bispecific antibodies).
  • ⁇ Attorney Docket No.250298.000954
  • the multispecific antigen-binding molecules are trispecific.
  • the multispecific antigen-binding molecules are tetraspecific.
  • the present disclosure includes multispecific (e.g., bispecific) antigen-binding molecules (e.g., antibodies) that specifically bind a capsid of an AAV particle and a molecule of a cell surface.
  • antigen-binding molecules may be referred to herein as, e.g., “anti- AAV x anti-cell surface molecule” or other similar terminology (e.g., anti-AAV/anti-cell surface molecule).
  • antigen-binding domain or “ABD” as used herein can refer to the portion of an antigen-binding molecule that is capable of specific binding to an antigen.
  • a multispecific antigen-binding molecule of the disclosure may comprise a first ABD (“ABD1”), a second ABD (“ABD2”), a third ABD (“ABD3”), and/or a fourth ABD (“ABD4”), each of which may be part of, e.g., an scFv or a Fab.
  • ABD1 ABD
  • BBD2 second ABD
  • ABS3 third ABD
  • ABS4 fourth ABD
  • antibody means any antigen-binding molecule or molecular complex comprising at least one complementarity determining region (CDR) that specifically binds to or interacts with a particular antigen.
  • CDR complementarity determining region
  • antibody includes immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, as well as multimers thereof (e.g., IgM).
  • antibody also includes immunoglobulin molecules consisting of four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds.
  • Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region.
  • the heavy chain constant region comprises three domains, CH1, CH2, and CH3.
  • Each light chain comprises a light chain variable region (abbreviated herein as LCVR or V L ) and a light chain constant region.
  • the light chain constant region comprises one domain (CL1).
  • CL1 The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR).
  • CDRs complementarity determining regions
  • FR framework regions
  • Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • the FRs of the anti-AAV antibody or anti-cell surface molecule antibody (or antigen-binding portion thereof) may be identical to the human germline sequences or may be naturally or artificially modified.
  • an amino acid consensus sequence may be defined based on a side-by-side analysis of two or more CDRs.
  • antibody also includes antigen-binding fragments of full antibody molecules.
  • antigen-binding fragment of an antibody, “antigen-binding portion” of an antibody, and the like, as used herein, include any naturally occurring, ⁇ ⁇ Attorney Docket No.250298.000954 enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex.
  • Antigen-binding fragments of an antibody may be derived, e.g., from full antibody molecules using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable and optionally constant domains.
  • DNA is known and/or is readily available from, e.g., commercial sources, DNA libraries (including, e.g., phage-antibody libraries), or can be synthesized.
  • the DNA may be sequenced and manipulated chemically or by using molecular biology techniques, for example, to arrange one or more variable and/or constant domains into a suitable configuration, or to introduce codons, create cysteine residues, modify, add, or delete amino acids, etc.
  • Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a constrained FR3- CDR3-FR4 peptide.
  • CDR complementarity determining region
  • an antigen-binding fragment of an antibody will typically comprise at least one variable domain.
  • the variable domain may be of any size or amino acid composition and will generally comprise at least one CDR which is adjacent to or in frame with one or more framework sequences.
  • the VH and VL domains may be situated relative to one another in any suitable arrangement.
  • the variable region may be dimeric and contain V H -V H , V H -V L , or V L -V L dimers.
  • the antigen-binding fragment of an antibody may contain a monomeric VH or VL domain.
  • an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain.
  • Non-limiting, exemplary configurations of variable and constant domains that may be found within an antigen-binding fragment of an antibody of the present disclosure include: (i) V H -C H 1; (ii) V H - CH2; (iii) VH-CH3; (iv) VH-CH1-CH2; (v) VH-CH1-CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL- CH1; (ix) VL-CH2; (x) VL-CH3; (xi) VL-CH1-CH2; (xii) VL-CH1-CH2-CH3; (xiii) VL-CH2-CH3; and ⁇ ⁇ Attorney Docket No.250298.000954 (xiv) V L -C L .
  • variable and constant domains may be either directly linked to one another or may be linked by a full or partial hinge or linker region.
  • a hinge region may consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids which result in a flexible or semi-flexible linkage between adjacent variable and/or constant domains in a single polypeptide molecule.
  • an antigen-binding fragment of an antibody of the present disclosure may comprise a homo-dimer or hetero-dimer (or other multimer) of any of the variable and constant domain configurations listed above in non- covalent association with one another and/or with one or more monomeric V H or V L domain (e.g., by disulfide bond(s)).
  • antigen-binding fragments may be monospecific or multispecific (e.g., bispecific).
  • a bispecific antigen-binding fragment of an antibody will typically comprise at least two different variable domains, wherein each variable domain is capable of specifically binding to a separate antigen or to a different epitope on the same antigen.
  • CDR complementarity determining region
  • Exemplary conventions that can be used to identify the boundaries of CDRs include, e.g., the Kabat definition, the Chothia definition, the ABS definition, and the IMGT definition. See, e.g., Kabat, 1991, “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Md. (Kabat numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol.273:927-948 (Chothia numbering scheme); Martin et al., 1989, Proc. Natl. Acad. Sci. USA 86:9268-9272 (ABS numbering scheme); and Lefranc et ai, 2003, Dev. Comp.
  • single domain antibody or “sdAb” as used herein can refer to an antibody or antigen binding fragment thereof comprising a single binding domain (e.g., heavy chain variable region) capable of binding a target molecule without pairing with a corresponding CDR-containing polypeptide (e.g., a light chain).
  • An sdAb or sdAb fragment can be derived from a VHH or from a non-antibody scaffold protein, for example a designed ankyrin repeat protein (darpin), an avimer, an anticalin/lipocalin, a centyrin, or a fynomer.
  • a ⁇ ⁇ Attorney Docket No.250298.000954 sdAb typically lacks a CH1 domain and thus cannot associate with a light chain.
  • VHH refers to a variable region of an antibody consisting of only a heavy chain, e.g., an antibody of camelid or cartilaginous fish origin. A VHH variable region can bind to a target molecule in the absence of a light chain.
  • a basic VHH has the following structure from the N-terminus to the C-terminus: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, in which FR1 to FR4 refer to framework regions 1 to 4, respectively, and in which CDR1 to CDR3 refer to the complementarity determining regions 1 to 3.
  • FR1 to FR4 refer to framework regions 1 to 4, respectively, and in which CDR1 to CDR3 refer to the complementarity determining regions 1 to 3.
  • the term “single chain Fv” or “scFv” as used herein can refer to a polypeptide chain comprising the V H and V L domains of an antibody, where these domains are present in a single polypeptide chain.
  • Fab can refer to a pair of polypeptide chains, the first polypeptide chain comprising a variable heavy (V H ) domain of an antibody N- terminal to a first constant domain (referred to herein as C1), and the second polypeptide chain comprising a variable light (VL) domain of an antibody N-terminal to a second constant domain (referred to herein as C2) capable of pairing with the first constant domain.
  • V H variable heavy
  • VL variable light domain of an antibody N-terminal to a second constant domain
  • the Fabs of the disclosure can be arranged according to the native orientation or include domain substitutions or swaps on that facilitate correct VH and VL pairings, particularly where the antigen-binding molecules of the disclosure comprise non-identical Fabs.
  • universal light chain as used herein in the context of an antigen-binding molecule described herein can refer to a light chain polypeptide capable of pairing with the heavy chain region of a first Fab to form the first Fab and capable of pairing with the heavy chain region of a second Fab to form the second Fab. Universal light chains are also known as “common light chains.”
  • Fc domain can refer to a portion of the heavy chain that pairs with the corresponding portion of another heavy chain.
  • Fc region can refer to the region of antibody-based binding molecules formed by association of two heavy chain Fc domains.
  • the two Fc domains within the Fc region may be the same or different from one another.
  • ⁇ ⁇ Attorney Docket No.250298.000954 a native antibody the Fc domains are typically identical, but for the purpose of producing the antigen-binding molecules of the disclosure, one or both Fc domains might advantageously be modified to allow for heterodimerization, e.g., via a knob-in-hole interaction and/or for purification, e.g., via star mutations.
  • the term “derived from” indicates a relationship between a first and a second molecule.
  • the term “specifically binds” as used herein means that an antigen-binding molecule forms a complex with a target antigen that is relatively stable under physiologic conditions.
  • Specific binding can be characterized by a KD of about 5x10 -2 M or less (e.g., less than 5x10 -2 M, less than 10 -2 M, less than 5x10 -2 M, less than 10 -3 M, less than 5x10 -4 M, less than 10 -4 M, less than 5x10 -5 M, less than 10 -5 M, less than 5x10 -6 M, less than 10 -6 M, less than 5x10 -7 M, less than 10 -7 M, less than 5x10 -8 M, less than 10 -8 M, less than 5x10 -9 M, less than 10 -9 M, or less than 10 -10 M).
  • a KD of about 5x10 -2 M or less (e.g., less than 5x10 -2 M, less than 10 -2 M, less than 5x10 -2 M, less than 10 -3 M, less than 5x10 -4 M, less than 10 -4 M, less than 5x10 -5 M, less than 10 -5 M, less than 5x10
  • an antigen-binding molecule or antigen-binding domain e.g., an antigen-binding molecule or antigen-binding domain
  • a target antigen e.g., an antigen-binding molecule or antigen-binding domain
  • an antigen-binding molecule that specifically binds to a target antigen from one species can, however, have cross-reactivity to the target antigen from one or more other species.
  • operably linked can refer to a functional relationship between two or more regions of a polypeptide chain in which the two or more regions are linked so as to produce a functional polypeptide.
  • substantially identical when referring to a nucleic acid or fragment thereof, indicates that, when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 90%, and more preferably at least about 95%, 96%, 97%, 98%, or 99% of the nucleotide bases, as measured by any well- known algorithm of sequence identity, such as FASTA, BLAST, or Gap, as discussed below.
  • a nucleic acid molecule having substantial identity to a reference nucleic acid molecule may, in certain instances, encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.
  • the term “substantial similarity” or “substantially similar” means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least 95% sequence identity, even more ⁇ ⁇ Attorney Docket No.250298.000954 preferably at least 98% or 99% sequence identity.
  • residue positions which are not identical differ by conservative amino acid substitutions.
  • a “conservative amino acid substitution” is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity).
  • R group side chain
  • a conservative amino acid substitution will not substantially change the functional properties of a protein.
  • the percent sequence identity or degree of similarity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well-known to those of skill in the art. See, e.g., Pearson (1994) Methods Mol. Biol.24: 307-331, herein incorporated by reference.
  • Examples of groups of amino acids that have side chains with similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic-hydroxyl side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartate and glutamate, and (7) sulfur-containing side chains are cysteine and methionine.
  • Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine- tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine.
  • a conservative replacement is any change having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256: 1443-1445, herein incorporated by reference.
  • a “moderately conservative” replacement is any change having a nonnegative value in the PAM250 log-likelihood matrix.
  • Protein analysis software matches similar sequences using measures of similarity assigned to various substitutions, deletions, and other modifications, including conservative amino acid substitutions.
  • GCG software contains programs such as Gap and Bestfit which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild type protein and a mutein thereof. See, e.g., GCG Version 6.1.
  • Polypeptide sequences also can be compared using FASTA using default or recommended parameters, a program in GCG Version 6.1.
  • FASTA e.g., FASTA2 and FASTA3 provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson (2000) supra).
  • Another preferred algorithm when comparing a sequence of the disclosure to a database containing a large number of sequences from different organisms is the computer program BLAST, especially BLASTP or TBLASTN, using ⁇ ⁇ Attorney Docket No.250298.000954 default parameters. See, e.g., Altschul et al. (1990) J. Mol. Biol.215:403-410 and Altschul et al. (1997) Nucleic Acids Res.25:3389-402, each herein incorporated by reference. [00330]
  • the term “subject” or “patient” as used herein includes all members of the animal kingdom including non-human primates and humans.
  • antibodies and antigen-binding fragments thereof that bind a capsid of an AAV particle also known as “anti-AAV antibodies” herein.
  • the capsid comprises any of various wild-type and/or non-wild-type AAV capsid protein(s) described herein.
  • the antibodies and/or antigen- binding fragments thereof bind an epitope on a capsid described herein.
  • AAV is an abbreviation for adeno-associated virus and may be used to refer to the virus itself or derivatives thereof.
  • AAVs are members of the Parvovirus family of small, non- enveloped, single-stranded DNA viruses. Generally, a wildtype AAV genome is 4.7 kb and is characterized by two inverted terminal repeats (ITR) and two open reading frames (ORFs), rep and cap.
  • the wildtype rep reading frame encodes four proteins of molecular weight 78 kD (“Rep78”), 68 kD (“Rep68”), 52 kD (“Rep52”) and 40 kD (“Rep 40”).
  • Rep78 and Rep68 are transcribed from the p5 promoter
  • Rep52 and Rep40 are transcribed from the p19 promoter. These proteins function mainly in regulating the transcription and replication of the AAV genome.
  • the wildtype cap reading frame encodes three structural (capsid) viral proteins (VPs) having molecular weights of 83-85 kD (VP1), 72-73 kD (VP2) and 61-62 kD (VP3).
  • More than 80% of total proteins in an AAV virion (capsid) comprise VP3; in mature virions VP1, VP2, and VP3 are found at relative abundance of approximately 1:1:10, although ratios of 1:1:8 have been reported. Padron et al. (2005) J. Virology 79:5047-58.
  • the genomic sequences of various serotypes of AAV, as well as the sequences of the native inverted terminal repeats (ITRs), Rep proteins, and capsid subunits are known in the art. Such sequences may be found in the literature or in public databases such as GenBank.
  • AAV encompasses all subtypes and both naturally occurring and modified forms (e.g., recombinant forms), except where stated otherwise.
  • AAV includes primate AAV (e.g., AAV type 1 (AAV1), primate AAV type 2 (AAV2), primate AAV type 3 (AAV3), primate AAV3B, primate AAV type 4 (AAV4), primate AAV type 5 (AAV5), primate AAV type 6 (AAV6), primate AAV6.2, primate AAV type 7 (AAV7), primate AAV type 8 (AAV8), primate AAV type 9 (AAV9), AAV10, AAV type hu11 (AAV hu11), AAV11, AAV12, AAV13, AAVDJ, Anc80L65, AAV2G9, AAVLK03, AAV type rh32.33 (AAVrh.32.33), AAV retro (AAV retro), AAV PHP.B, AAV PHP.eB, AAV PHP.S, AAVrh.64R1, AAVhu.37, AAVrh.8, AAV2/8, etc.; non-prim
  • non- primate animal AAV can refer to AAV isolated from non-primate animals.
  • capsid protein includes a protein that is part of the capsid of the virus.
  • the capsid proteins are generally referred to as VP1, VP2, and/or VP3, and may be encoded by the single cap gene.
  • the three AAV capsid proteins can be produced in nature an overlapping fashion from the cap ORF alternative translational start codon usage, although all three proteins use a common stop codon.
  • the ORF of a wildtype cap gene encodes from 5’ to 3’ three alternative start codons: “the VP1 start codon,” “the VP2 start codon,” and “the VP3 start codon”; and one “common stop codon”.
  • the largest viral protein, VP1 is generally encoded from the VP1 start codon to the “common stop codon.”
  • VP2 is generally encoded from the VP2 start codon to the common stop codon.
  • VP3 is generally encoded from the VP3 start codon to the common stop codon.
  • VP1 comprises at its N-terminus sequence that it does not share with the VP2 or VP3, referred to as the VP1-unique region (VP1-u).
  • the VP1-u region is generally encoded by the sequence of a wildtype cap gene starting from the VP1 start codon to the “VP2 start codon.”
  • VP1-u comprises a phospholipase A2 domain (PLA2), which may be important for infection, as well as nuclear localization signals which may aid the virus in targeting to the nucleus for uncoating and genome release.
  • PHA2 phospholipase A2 domain
  • the VP1, VP2, and VP3 capsid proteins share the same C-terminal sequence that makes up the ⁇ ⁇ Attorney Docket No.250298.000954 entirety of VP3, which may also be referred to herein as the VP3 region.
  • the VP3 region is encoded from the VP3 start codon to the common stop codon.
  • an antibody, or an antigen-binding fragment thereof that binds to a capsid of an AAV particle described may bind to an epitope on the capsid.
  • the antibody may comprise any of the antibodies, e.g., multispecific antibodies, such as bispecific antibodies, which bind to the capsid of an AAV particle disclosed herein.
  • a multispecific antibody disclosed herein may comprise, for example, comprise a first binding domain that binds to a capsid of an AAV particle.
  • the first binding domain binds to an epitope on the capsid of the AAV particle.
  • the AAV particle capsid may comprise wild-type and/or non-wildtype AAV capsid proteins.
  • the epitope may comprise any of various serotype- specific or serotype-non-specific (i.e., “universal”) epitopes understood by one of ordinary skill in the art.
  • the epitope may comprise, e.g., any amino acid residues set forth in Tables 19-26, or variants of variants, derivatives, or epitopes combinations thereof.
  • an antibody, or an antigen-binding fragment thereof that binds to a capsid of an AAV particle described herein may bind to an epitope on the capsid which is comprised of any amino acid residues of a capsid protein VP1, VP2, or VP3, or a combination thereof.
  • the AAV particle can be derived from any AAV serotype described herein.
  • an antibody, or an antigen-binding fragment thereof can bind to any amino acid residue, or any combination thereof, of any variable region of a capsid, or any combination thereof, e.g., a variable region comprised of a capsid protein VP1, VP2, or VP3, or a combination thereof (e.g., variable region I, II, III, IV, V, VI, VII, VIII, IX, or a combination thereof, as described in Emmanuel, et al., Journal of Virology, 2022, 96, 3).
  • an antibody, or an antigen-binding fragment thereof that binds to a capsid of an AAV particle described herein may bind to an epitope of an AAV1 capsid comprised of any amino acid residue or combination thereof of a variable region I, II, III, IV, V, VI, VII, VIII, IX, or combination thereof.
  • the antibody, or an antigen-binding fragment thereof may bind to AAV1 at any of residues 456–459, 492–499, 582, 583, 588– 591, 593–595, 597, or any combination thereof, as described in Tseng, et al., ⁇ Front Immunol. 2014, 5: 9.
  • an antibody, or an antigen-binding fragment thereof that binds to a capsid of an AAV particle described herein may bind to an epitope of an AAV2 capsid comprised of any amino acid residue or combination thereof of a variable region I, II, III, IV, V, VI, VII, VIII, IX, or combination thereof.
  • the antibody, or an antigen-binding ⁇ ⁇ Attorney Docket No.250298.000954 fragment thereof may bind to AAV2 at any of residues 253, 254, 258, 261-264, 272–281, 369–378, 381, 384, 385, 474–483, 492–502, 534, 548, 556, 560–573, 585–589, 601–610, 658–660, 708, 717, or any combination thereof, as described in Tseng, et al., ⁇ Front Immunol. 2014, 5: 9.
  • an antibody, or an antigen-binding fragment thereof that binds to a capsid of an AAV particle described herein may bind to an epitope of an AAV5 capsid comprised of any amino acid residue or combination thereof of a variable region I, II, III, IV, V, VI, VII, VIII, IX, or combination thereof.
  • the antibody, or an antigen-binding fragment thereof may bind to AAV5 at any of residues 246, 254–261, 374, 375, 483, 485– 492, 494, 496, 499-501, 530, 532–538, 653, 654, 656, 657, 704–708, or any combination thereof as described in Tseng, et al., ⁇ Front Immunol.2014, 5: 9.
  • an antibody, or an antigen-binding fragment thereof that binds to a capsid of an AAV particle described herein may bind to an epitope of an AAV8 capsid comprised of any amino acid residue or combination thereof of a variable region I, II, III, IV, V, VI, VII, VIII, IX, or combination thereof.
  • the antibody, or an antigen-binding fragment thereof may bind to AAV8 at any of residues 586–591 or any combination thereof as described in Tseng, et al., ⁇ Front Immunol.2014, 5: 9.
  • an antibody, or an antigen-binding fragment thereof that binds to a capsid of an AAV particle described herein may bind to an epitope of an AAV9 capsid comprised of any amino acid residue or combination thereof of a variable region I, II, III, IV, V, VI, VII, VIII, IX, or combination thereof.
  • the antibody, or an antigen-binding fragment thereof may bind to AAV9 at any of residues 221, 228, 246-248, 250-256, 258– 260, any of residues 262–273 of variable region I, any of residues 274, 275, 278, 291, 293, 324, 325, any of residues 327-333 of variable region II, any of residues 334, 335, 338, 341, 363-365, 369-373, 375, 376, any of residues 382-387, 389, 391 of variable region III, any of residues 438–441, 443, any of residues 446, 448, 449, 451-460, 462, 464-473 of variable region IV, any of residues 488, 491-501, 503-506, 510–515 of variable region V, any of residues 528-534, 545 of variable region VI, any of residues 547-560 of variable region VII, residue 572, any of residues 579, 584, 587–590 of
  • a Cap protein e.g., a VP1 capsid protein as described herein, a VP2 capsid protein as described herein, and/or a VP3 capsid protein as described herein, can be modified to comprise any number of various capsid modifications, e.g., one or more of a point mutation (e.g., amino acid substitutions, deletions, and/or insertions), a ⁇ ⁇ Attorney Docket No.250298.000954 detectable label, a member of a protein:protein binding pair, etc.
  • a point mutation e.g., amino acid substitutions, deletions, and/or insertions
  • a ⁇ ⁇ Attorney Docket No.250298.000954 detectable label
  • a member of a protein:protein binding pair etc.
  • the one or more point mutation(s) in a capsid protein can result in detargeting a viral particle comprising the mutant capsid protein from its natural target cell.
  • Detargeting of a viral particle from its natural target cell can be important especially if systemic versus local or loco-regional administration of the viral particles is intended, as uptake of the viral particles by the natural target cell can limit the effective dose of the viral particles.
  • point mutations can be those that reduce the transducing activity of a viral particle which is mediated by the natural receptor of the viral particle by at least 50%, preferably at least 80%, especially at least 95%.
  • detargeting mutations disclosed herein can involve deletion or replacement of one or more an amino acids which can be involved in binding of the respective viral particle to its natural receptor. Such point mutations can enable an efficient detargeting of the viral particle from cells expressing the natural receptor or, for targeting purposes, can increase specificity of the respective mutant viral particle for a new target cell.
  • a capsid protein e.g., a VP1, VP2, and/or VP3 capsid protein
  • a capsid protein can be modified to comprise an inserted heterologous amino acid sequence (e.g., a peptide insertion described herein or an additional peptide insertion, such as a peptide comprising, consisting essentially of, or consisting of any of various targeting molecules described herein) capable of retargeting or redirecting a viral particle described herein.
  • an inserted heterologous amino acid sequence e.g., a peptide insertion described herein or an additional peptide insertion, such as a peptide comprising, consisting essentially of, or consisting of any of various targeting molecules described herein
  • Retargeting may include a scenario in which a wild-type viral particle targets several cells within a tissue and/or several organs within an organism, and general targeting of the cells, tissues, and/or organs is reduced or abolished by insertion of a heterologous amino acid sequence, and retargeting the viral particle to more a specific cell in the tissue or a specific organ in the organism is achieved with the inserted heterologous amino acid sequence which can, e.g., bind a marker expressed by a specific cell.
  • Such retargeting or redirecting may also include a scenario in which the wild- type viral particle targets a tissue, and targeting of the tissue is reduced or abolished by insertion of the heterologous amino acid sequence, and retargeting to a completely different tissue is achieved with the inserted heterologous amino acid sequence.
  • One or more of detargeting and/or retargeting mutation(s) and/or insertion(s) can be introduced into capsid proteins of any of various AAV serotypes disclosed herein.
  • Such detargeting and/or retargeting mutation(s) and/or insertion(s) can result in detargeting and/or retargeting of an AAV particle comprising a mutant capsid protein from or to specific cells within a tissue and/or organs within an organism.
  • cells, tissues, and/or organs which may be detargeted and/or detargeted by AAV particles comprising mutant capsids ⁇ ⁇ Attorney Docket No.250298.000954 may include or be derived from, without limitation, liver, skeletal muscle, vascular and/or smooth muscle, cardiac muscle, nervous system (e.g., brain, spinal cord, neurons, glia, ependymal cells), eye (e.g., retina and retinal cells including photoreceptors such as rods and cones, RPE, etc.), lung, heart, pancreas, kidney, and epithelium.
  • an AAV2 serotype is preferred for use in the compositions and/or methods of the present disclosure.
  • the capsid of the AAV2 particle can comprise one or more amino acid mutations within a heparan sulfate proteoglycan (HSPG) binding domain of the AAV2 capsid.
  • the capsid of the AAV2 particle can comprise one or more amino acid mutations within a laminin receptor (LamR) binding domain, e.g., a 37/67-kDa LamR binding domain, of the AAV2 capsid.
  • LamR laminin receptor
  • the capsid of the AAV2 particle can comprise one or more mutations at an amino acid position(s) selected from amino acid position(s) R484, R487, R585, R588, and K532 (VP1 numbering), and a combination thereof, relative to a wild-type AAV2 capsid.
  • the capsid of the AAV2 particle comprises a mutation at amino acid position R585 and/or R588.
  • the capsid of the AAV2 particle comprises one or more amino acid substitutions at an amino acid position(s) selected from R484A, R487A, R487G, K532A, K532D, R585A, R585S, R585Q, R588A, and R588T, and any combination thereof, relative to a wild-type AAV2 capsid.
  • the capsid of the AAV2 particle comprises an amino acid substitution R585A and/or R588A.
  • AAV2 R585 corresponds to AAV9 S586 and AAV8 Q588
  • AAV2 R588 corresponds to AAV9 A589 and AAV8 T591.
  • the capsid of the AAV9 particle can comprise a mutation at amino acid position S586 and/or A589.
  • the capsid of the AAV8 particle can comprise a mutation at amino acid position Q588 and/or T591.
  • the capsid of the AAV2 particle comprises one or more insertion(s) at amino acid position G453 or N587, or a combination thereof.
  • an AAV9 serotype is used in the compositions and/or methods of the present disclosure.
  • the capsid of the AAV9 particle can comprise one or more amino acid mutations within a galactose binding domain of the AAV9 capsid.
  • the capsid of the AAV9 particle can comprise one or more mutations at an amino acid position(s) selected from amino acid position(s) D271, N272, Y446, S469, N470, A742, V473, W503, E500, P504, and Q590 (VP1 numbering), and a combination thereof, relative to a wild-type AAV9 capsid.
  • the capsid of the AAV9 particle comprises a mutation at amino acid position N272 and/or W503.
  • the capsid of the AAV9 particle comprises one or more amino acid substitutions selected from K532A, K532D, R484A, R487A, R585A, R585S, R585Q, R487G, ⁇ ⁇ Attorney Docket No.250298.000954 R588A, and R588T, and any combination thereof, relative to a wild-type AAV2 capsid.
  • the capsid of the AAV9 particle comprises an amino acid substitution N272A and/or W503A, relative to a wild-type AAV9 capsid.
  • the capsid of the AAV9 particle comprises one or more insertion(s) at amino acid position G453, A587, or A589, or a combination thereof.
  • an AAV1 serotype is used in the compositions and/or methods of the present disclosure.
  • the capsid of the AAV1 particle can comprise one or more amino acid mutations within a sialic acid binding domain, e.g., an N-linked sialic acid binding domain, of the AAV1 capsid.
  • the capsid of the AAV1 particle can comprise one or more amino acid mutations within a binding domain of the AAV1 capsid essential for binding adeno-associated virus receptor (AAVR).
  • AAVR adeno-associated virus receptor
  • the capsid of the AAV1 particle can comprise one or more mutations at an amino acid position(s) selected from amino acid position(s) N500 (which corresponds to AAV2 E499 and AAV9 E500), K531, and K531 (VP1 numbering), and a combination thereof, relative to a wild-type AAV1 capsid.
  • the capsid of the AAV1 particle comprises one or more amino acid substitutions selected from N500E, K531A, and K531E, and any combination thereof, relative to a wild-type AAV1 capsid.
  • an AAV6 serotype is used is used in the compositions and/or methods of the present disclosure.
  • the capsid of the AAV6 particle can comprise one or more amino acid mutations within a sialic acid binding domain, e.g., an N-linked sialic acid binding domain, of the AAV6 capsid. In some embodiments, the capsid of the AAV6 particle can comprise one or more amino acid mutations within a heparan sulfate proteoglycan (HSPG) binding domain of the AAV6 capsid. In some embodiments, the capsid of the AAV6 particle can comprise one or more amino acid mutations within an epidermal growth factor receptor (EGFR) binding domain of the AAV6 capsid.
  • EGFR epidermal growth factor receptor
  • the capsid of the AAV6 particle can comprise an amino acid mutation at amino acid position N500 (which corresponds to AAV2 E499 and AAV9 E500) (VP1 numbering), relative to a wild-type AAV6 capsid.
  • the capsid of the AAV6 particle comprises amino acid substitution N500E relative to a wild-type AAV6 capsid.
  • the capsid of the AAV6 particle comprises an insertion at amino acid position Q585.
  • an AAV8 serotype is used is used in the compositions and/or methods of the present disclosure.
  • the capsid of the AAV8 particle can comprise one or more amino acid mutations within a laminin receptor (LamR) binding domain, e.g., a 37/67-kDa LamR binding domain, of the AAV8 capsid.
  • the capsid of the AAV8 particle comprises an insertion at amino acid position ⁇ ⁇ Attorney Docket No.250298.000954 N590 (VP1 numbering).
  • an AAV5 serotype is used in the compositions and/or methods of the present disclosure.
  • the capsid of the AAV5 particle can comprise one or more amino acid mutations within a sialic acid binding domain, e.g., an N-linked sialic acid binding domain, of the AAV5 capsid. In some embodiments, the capsid of the AAV5 particle can comprise one or more amino acid mutations within a platelet-derived growth factor receptor (PDGFR) of the AAV5 capsid. In some embodiments, the capsid of the AAV5 particle can comprise a mutation at amino acid position T571 (VP1 numbering) relative to a wild-type AAV5 capsid.
  • PDGFR platelet-derived growth factor receptor
  • the capsid of the AAV5 particle comprises an amino acid substitution T571S relative to a wild-type AAV5 capsid.
  • an avian AAV is used in the compositions and/or methods of the present disclosure.
  • the capsid of the avian AAV particle comprises an insertion at amino acid position(s) G444 or K580 (VP1 numbering), or a combination thereof.
  • an bearded dragon AAV is used in the compositions and/or methods of the present disclosure.
  • the capsid of the bearded dragon AAV particle comprises an insertion at amino acid position(s) G436 or T573 (VP1 numbering), or a combination thereof.
  • ITR inverted terminal repeat
  • the phrase “inverted terminal repeat” or “ITR” includes symmetrical nucleic acid sequences in the genome of adeno-associated viruses required for efficient replication. ITR sequences are located at each end of the AAV DNA genome. The ITRs serve as the origins of replication for viral DNA synthesis and are essential cis components for generating AAV particles, e.g., packaging into AAV particles.
  • AAV ITR comprise recognition sites for replication proteins Rep78 or Rep68.
  • a “D” region of the ITR comprises the DNA nick site where DNA replication initiates and provides directionality to the nucleic acid replication step.
  • An AAV replicating in a mammalian cell typically comprises two ITR sequences.
  • a single ITR may be engineered with Rep binding sites on both strands of the “A” regions and two symmetrical D regions on each side of the ITR palindrome. Such an engineered construct on a double-stranded circular DNA template allows Rep78 or Rep68 initiated nucleic acid replication that proceeds in both directions.
  • a single ITR is sufficient for AAV replication of a circular particle.
  • the rep encoding sequence encodes a Rep protein or Rep protein equivalent that is capable of binding an ITR comprised on the transfer plasmid.
  • the Cap proteins of the disclosure when expressed with appropriate Rep proteins by a packaging cell, may encapsidate a transfer plasmid comprising a nucleotide of interest ⁇ ⁇ Attorney Docket No.250298.000954 and an even number of two or more ITR sequences.
  • a transfer plasmid comprises one ITR sequence.
  • a transfer plasmid comprises two ITR sequences.
  • Rep proteins may be expressed from more than one ORF comprising nucleotide sequence encoding any combination of Rep78, Rep68, Rep 52, and/or Rep40 by use of separate nucleotide sequences operably linked to at least one expression control sequence for expression in a viral replication cell, each producing one or more of Rep78, Rep68, Rep 52, and/or Rep40 Rep proteins.
  • Rep proteins may be expressed individually from an ORF comprising a nucleotide sequence encoding any one of Rep78, Rep68, Rep 52, or Rep40 by use of separate nucleotide sequences operably linked to one expression control sequence for expression in a packaging cell, each producing only one Rep78, Rep68, Rep 52, or Rep40 Rep protein.
  • Rep proteins may be expressed from one ORF comprising nucleotide sequences encoding Rep78 and Rep52 Rep proteins operably linked to at least one expression control sequence for expression in a viral replication cell each producing Rep78 and Rep52 Rep protein.
  • a “chimeric AAV capsid protein” includes an AAV capsid protein that comprises amino acid sequences, e.g., portions, from two or more different AAV and that is capable of forming and/or forms an AAV viral capsid/viral particle.
  • a chimeric AAV capsid protein is encoded by a chimeric AAV capsid gene, e.g., a chimeric nucleotide comprising a plurality, e.g., at least two, nucleic acid sequences, each of which plurality is identical to a portion of a capsid gene encoding a capsid protein of distinct AAV, and which plurality together encodes a functional chimeric AAV capsid protein.
  • a chimeric capsid protein comprises one or more portions from a capsid protein of that AAV and one or more portions from a capsid protein of a different AAV.
  • a chimeric AAV2 capsid protein includes a capsid protein comprising one or more portions of a VP1, VP2, and/or VP3 capsid protein of AAV2 and one or more portions of a VP1, VP2, and/or VP3 capsid protein of a different AAV.
  • Table 1 sets forth the amino acid sequence identifiers of the heavy chain variable regions (HCVRs) and light chain variable regions (LCVRs), heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3), and light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), as well as heavy chain (HC) and light chain (LC) of the exemplary anti-AAV antibodies and antigen- ⁇ ⁇ Attorney Docket No.250298.000954 binding fragments.
  • Table 2 sets forth the sequence identifiers of the nucleic acid molecules encoding the HCVRs, LCVRs, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 LCDR3, HC, and LC of the exemplary anti-AAV antibodies and antigen-binding fragments described herein.
  • Corresponding Fab and “standard mAb” i.e., mAb comprising a constant region derived from IgG4 subclass
  • Non-limiting examples of amino acid sequences and nucleotide sequences of the anti-AAV antibodies are also listed below. Table 1.
  • anti-AAV antibodies, or antigen-binding fragments thereof comprising an LCVR comprising an amino acid sequence selected from any of the LCVR amino acid sequences listed in Table 1, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • anti-AAV antibodies, or antigen-binding fragments thereof comprising an HCVR and an LCVR amino acid sequence pair (HCVR/LCVR) comprising any of the HCVR amino acid sequences listed in Table 1 paired with any of the LCVR amino acid sequences listed in Table 1.
  • the present disclosure provides antibodies, or antigen-binding fragments thereof, comprising an HCVR/LCVR amino acid sequence pair contained within any of the exemplary anti-AAV antibodies listed in Table 1.
  • the HCVR/LCVR amino acid sequence pair is selected from SEQ ID NOs: 2/10 (e.g., REGN13876); 22/10 (e.g., REGN13877); 32/10 (e.g., REGN13878); 42/10 (e.g., REGN13879); 52 or 1159/10 or 1157 (e.g., REGN13880); 62/10 (e.g., REGN13881); 71/10 (e.g., REGN13882); 81/10 (e.g., REGN13883); 91/10 (e.g., REGN13884); 101/10 (e.g., REGN13885); 111/119 (e.g., REGN13070); 131/119 (e.g., REGN13071); 141/119 (e.g., REGN13885); 111/119
  • anti-AAV antibodies, or antigen-binding fragments thereof comprising a heavy chain CDR1 (HCDR1) comprising an amino acid sequence selected from any of the HCDR1 amino acid sequences listed in Table 1 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • anti-AAV antibodies, or antigen-binding fragments thereof comprising a heavy chain CDR2 (HCDR2) comprising an amino acid sequence selected from any of the HCDR2 amino acid sequences listed in Table 1 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • anti-AAV antibodies, or antigen-binding fragments thereof comprising a heavy chain CDR3 (HCDR3) comprising an amino acid sequence selected from any of the HCDR3 amino acid sequences listed in Table 1 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • anti-AAV antibodies, or antigen-binding fragments thereof comprising a light chain CDR1 (LCDR1) comprising an amino acid sequence selected from any of the LCDR1 amino acid sequences listed in Table 1 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • LCDR1 light chain CDR1
  • anti- AAV antibodies, or antigen-binding fragments thereof comprising a light chain CDR2 (LCDR2) comprising an amino acid sequence selected from any of the LCDR2 amino acid sequences listed in Table 1 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • LCDR2 light chain CDR2
  • LCDR3 light chain CDR3
  • the anti-AAV antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 18, 198, 30, 208, 40, 127, 50, 169, 60, 1108, 1363, 1366, 149, 1362, 1379, 69, 178, 79, 1391, 1396, 188, 1390, 89, 139, 99, 159, 109, or 218, or a variant thereof having at least 40%, ⁇ ⁇ Attorney Docket No.250298.000954 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 18, 198, 30, 208, 40, 127, 50, 169, 60, 1108, 1363, 1366, 149, 1362, 1379, 69, 178, 79, 1391, 1396, 188, 1390, 89,
  • the nucleotide sequence that encodes the anti- AAV antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 18, 198, 30, 208, 40, 127, 50, 169, 60, 1108, 1363, 1366, 149, 1362, 1379, 69, 178, 79, 1391, 1396, 188, 1390, 89, 139, 99, 159, 109, or 218, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 18, 198, 30, 208, 40, 127, 50, 169, 60, 1108, 1363, 1366, 149, 1362, 1379, 69, 178, 79, 1391, 1396, 188, 1390, 89, 139, 99,
  • the nucleotide sequence that encodes the anti-AAV antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 17, 197, 29, 207, 39, 126, 49, 168, 59, 1107, 148, 1378, 68, 177, 78, 187, 1389, 88, 138, 98, 158, 108, or 217, or a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the nucleotide sequence of SEQ ID NOs: 17, 197, 29, 207, 39, 126, 49, 168, 59, 1107, 148, 1378, 68, 177, 78, 187, 1389, 88, 138, 98, 158, 108, or 217.
  • the anti-AAV antibodies, or antigen- binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NOs: 18, 198, 30, 208, 40, 127, 50, 169, 60, 1108, 1363, 1366, 149, 1362, 1379, 69, 178, 79, 1391, 1396, 188, 1390, 89, 139, 99, 159, 109, or 218.
  • the nucleotide sequence that encodes the anti-AAV antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 17, 197, 29, 207, 39, 126, 49, 168, 59, 1107, 148, 1378, 68, 177, 78, 187, 1389, 88, 138, 98, 158, 108, or 217.
  • the anti-AAV antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 20, 129, or 813, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 20, 129, or 813.
  • the nucleotide sequence that encodes the anti-AAV antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 20, 129, or 813, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 20, 129, or 813.
  • the nucleotide sequence that encodes the anti-AAV antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 19, 128, 1145, or 1500, or a nucleotide sequence having ⁇ ⁇ Attorney Docket No.250298.000954 at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the nucleotide sequence of SEQ ID NOs: 19, 128, 1145, or 1500.
  • the anti-AAV antibodies, or antigen-binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NOs: 20, 129, or 813.
  • the nucleotide sequence that encodes the anti-AAV antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 19, 128, 1145, or 1500.
  • antibodies, or antigen-binding fragments thereof comprising a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3) contained within any of the exemplary anti-AAV antibodies listed in Table 1.
  • the HCDR1- HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set is selected from SEQ ID NOs: 4-6-8-12-14-16 (e.g., REGN13876); 24-26-28-12-14-16 (e.g.-REGN13877); 34-36-38- 12-14-16 (e.g., REGN13878); 44-46-48-12-14-16 (e.g., REGN13879; 54-56-58-12-14-16 (e.g., REGN13880); 64-56-67-12-14-16 (e.g., REGN13881); 73-75-77-12-14-16 (e.g., REGN13882); 83-85-87-12-14-16 (e.g., REGN13883); 93-95-97-12-14-16 (e.g.- REGN13884); 103-105-107-12-14-16 (e.g., REGN13885); 113-115-117-121-123-125 (e.g.- REGN
  • antibodies, or antigen-binding fragments thereof comprising a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1- LCDR2-LCDR3) contained within an HCVR/LCVR amino acid sequence pair as defined by any of the exemplary anti-AAV antibodies listed in Table 1.
  • antibodies, or antigen-binding fragments thereof comprising the HCDR1-HCDR2-HCDR3- LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR/LCVR amino acid sequence pair selected from SEQ ID NOs: 2/10 (e.g., REGN13876); 22/10 (e.g., REGN13877); 32/10 (e.g., REGN13878); 42/10 (e.g., REGN13879); 52 or 1159/10 or 1157 (e.g., REGN13880); 62/10 (e.g., REGN13881); 71/10 (e.g., REGN13882); 81/10 (e.g., REGN13883); 91/10 (e.g., REGN13884); 101/10 (e.g., REGN13885); 111/119 (e.g., REGN13070); 131/119 (e.g., REGN13071); 141/119 (e.g., REGN13072); 151/
  • the anti-AAV antibody or antigen-binding ⁇ ⁇ Attorney Docket No.250298.000954 fragment thereof can include: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a heavy chain variable region (HCVR) that comprises a HCVR1, HCDR2, and
  • the anti-AAV antibody or antigen-binding fragment thereof can include: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCD
  • the anti-AAV antibody or antigen-binding fragment thereof can include: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 or 1159, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; f
  • an anti-AAV Fab of the present disclosure includes: a) an HC that comprises the amino acid sequence of SEQ ID NO: 18, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ ID NO: 20, or a variant thereof; b) an HC that comprises the amino acid sequence of SEQ ID NO: 30, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ ID NO: 20, or a variant thereof; c) an HC that comprises the amino acid sequence of SEQ ID NO: 40, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ ID NO: 20, or a variant thereof; d) an HC that comprises the amino acid sequence of SEQ ID NO: 50, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ ID NO: 20, or a variant thereof; e) an HC that comprises the amino acid sequence of SEQ ID NO: 50, or a variant thereof; and/or an LC that comprises the amino acid sequence of
  • an anti-AAV Fab of the present disclosure includes: k) an HC that comprises the amino acid sequence of SEQ ID NO: 127, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ ID NO: 129, or a variant thereof; l) an HC that comprises the amino acid sequence of SEQ ID NO: 139, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ ID NO: 129, or a variant thereof; m) an HC that comprises the amino acid sequence of SEQ ID NO: 149, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ ID NO: 129, or a variant thereof; n) an HC that comprises the amino acid sequence of SEQ ID NO: 159, or a variant thereof; and/or an LC that comprises the amino acid sequence of SEQ
  • the anti-AAV antibodies or antigen-binding fragments thereof described herein may bind to a capsid of an AAV particle with a KD value of about 5x10 -6 M or less, such as about 10 -7 M or less, about 10 -8 M or less, such as about 10 -9 M or less when determined by, for instance, surface plasmon resonance (SPR) technology in a BIAcore instrument using the antigen as the ligand and the antibody, Ig, antibody-binding fragment, or Fc-containing protein as the analyte (or antiligand).
  • SPR surface plasmon resonance
  • FACS fluorescent- activated cell sorting
  • an anti-AAV antibody or antigen- binding fragment thereof described herein may bind to a capsid of an AAV particle with a KD value of about 5x10 -6 M to 1x10 -6 M (e.g., 1x10 -6 M, 1.5x10 -6 M, 2x10 -6 M, 3x10 -6 M, 4x10 -6 M, 5x10 -6 M), about 1x10 -6 M to 1x10 -7 M (e.g., 1x10 -7 M, 2x10 -7 M, 3x10 -7 M, 4x10 -7 M, 5x10 -7 M, 6x10 -7 M, 7x10 -7 M, 8x10 -7 M, 9x10 -7 M), about 1x10 -7 M to 1x10 -8 M (e.g., 1x10 -8 M, 2x10- 8 M, 3x10 -8 M, 4x10 -8 M, 5x10 -8 M, 6x10 -8 M, 7x10 -8 M,
  • the antibody or antigen-binding protein of the disclosure may bind to the predetermined antigen or cell surface molecule (receptor) having an affinity corresponding to value that is at least ten-fold lower than its affinity for binding to a non-specific antigen (e.g., BSA).
  • a non-specific antigen e.g., BSA
  • the affinity of an antibody corresponding to a K D value that is equal to or less than ten-fold lower than a non-specific antigen may be considered non-detectable binding, however such an antibody may be paired with a second antigen-binding domain for the production of a multispecific antibody of the disclosure.
  • K D can refer to the dissociation equilibrium constant of a particular antibody-antigen interaction, or the dissociation equilibrium constant of an antibody or antibody-binding fragment binding to an antigen.
  • K D can refer to the dissociation equilibrium constant of a particular antibody-antigen interaction, or the dissociation equilibrium constant of an antibody or antibody-binding fragment binding to an antigen.
  • binding affinity There is an inverse relationship between K D and binding affinity, therefore the smaller the K D value, the higher, i.e. stronger, the affinity.
  • the terms “higher affinity” or “stronger affinity” relate to a higher ability to form an interaction and therefore a smaller K D value
  • the terms “lower affinity” or “weaker affinity” relate to a lower ability to form an interaction and therefore a larger KD value.
  • a higher binding affinity (or KD) of a particular molecule e.g.
  • koff value Said value is also referred to as the koff value.
  • k a M-1 x sec-1 or 1/M/s
  • K A K A
  • the association equilibrium constant is obtained by dividing the k a by the k d .
  • the term “EC50” or “EC50” can refer to the half maximal effective concentration, which includes the concentration of an antibody which induces a response halfway between the baseline and maximum after a specified exposure time.
  • the EC 50 essentially represents the concentration of an antibody where 50% of its maximal effect is observed.
  • the EC50 value equals the concentration of an antibody of the disclosure that gives half-maximal binding to a capsid protein of an AAV particle (e.g., a wild-type or a non- wild-type AAV capsid protein(s) or to cells expressing a molecule on a cell surface described herein, as determined by e.g., a FACS binding assay.
  • nucleic acid molecules encoding anti-AAV antibodies or antigen-binding fragments thereof are also provided herein.
  • nucleic acid molecules encoding any of the HCDR1 amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCDR1 nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • nucleic acid molecules encoding any of the HCDR2 amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCDR2 nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • nucleic acid molecules encoding any of the HCDR3 amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCDR3 nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • nucleic acid molecules encoding any of the LCVR amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCVR nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • nucleic acid molecules encoding any of the LCDR1 amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCDR1 nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • nucleic acid molecules encoding any of the LCDR2 amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCDR2 nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • nucleic acid molecules encoding any of the LCDR3 amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCDR3 nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • nucleic acid molecules encoding an HCVR wherein the HCVR comprises a set of three CDRs (i.e., HCDR1-HCDR2-HCDR3), wherein the HCDR1-HCDR2- HCDR3 amino acid sequence set is as defined by any of the exemplary anti-AAV antibodies listed in Table 1.
  • nucleic acid molecules encoding an LCVR wherein the LCVR comprises a set of three CDRs (i.e., LCDR1-LCDR2-LCDR3), wherein the LCDR1-LCDR2- ⁇ ⁇ Attorney Docket No.250298.000954 LCDR3 amino acid sequence set is as defined by any of the exemplary anti-AAV antibodies listed in Table 1.
  • nucleic acid molecules encoding both an HCVR and an LCVR wherein the HCVR comprises an amino acid sequence of any of the HCVR amino acid sequences listed in Table 1, and wherein the LCVR comprises an amino acid sequence of any of the LCVR amino acid sequences listed in Table 1.
  • the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCVR nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto, and a polynucleotide sequence selected from any of the LCVR nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • the nucleic acid molecule encodes an HCVR and LCVR, wherein the HCVR and LCVR are both derived from the same anti-AAV antibody listed in Table 1.
  • nucleic acid molecules encoding any of the HC amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HC nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • the present disclosure provides nucleic acid molecules encoding any of the LC amino acid sequences listed in Table 1; in certain embodiments the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LC nucleic acid sequences listed in Table 2, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.
  • Provided herein are recombinant expression vectors capable of expressing one or more polypeptides of an anti-AAV antibody or antigen-binding fragment thereof.
  • recombinant expression vectors comprising any of the nucleic acid molecules mentioned above, i.e., nucleic acid molecules encoding any of the HCVR, LCVR, and/or CDR sequences as set forth in Table 1.
  • host cells into which such vectors have been introduced are also included within the scope of the present disclosure, as well as methods of producing the antibodies or portions thereof by culturing the host cells under conditions permitting production of the antibodies or antibody fragments and recovering the antibodies and antibody fragments so produced.
  • host cell as used herein can refer to cells into which a nucleic acid of the disclosure has been introduced.
  • the terms “host cell” and “recombinant host cell” are used interchangeably herein.
  • Typical host cells are eukaryotic host cells, such as mammalian host cells.
  • exemplary eukaryotic host cells include yeast and mammalian cells, for example vertebrate cells such as a mouse, rat, monkey, or human cell line, for example HKB11 cells, PER.C6 cells, HEK cells, or CHO cells.
  • the antigen-binding molecules (e.g., antibodies) of the present disclosure may be monospecific or multispecific (e.g., bispecific).
  • Multispecific antigen-binding molecules also referred to as “multispecific binding molecules” or “MBMs” herein
  • MBMs multispecific binding molecules
  • the multispecific antigen-binding molecules of the disclosure may comprise at least one antigen-binding domain that binds to a capsid of an AAV particle, and at least one antigen-binding domain that binds to a different target molecule.
  • the MBMs of the disclosure specifically bind to at least two different epitopes (and in some instances three or more different epitopes).
  • the at least two different epitopes can be on the same target molecule or different target molecules.
  • the MBMs of the disclosure specifically bind to two or more different target molecules.
  • the MBMs described herein may bind to one or more epitopes of a capsid of an AAV particle.
  • the MBMs described herein may bind to one or more epitopes of a molecule on a cell surface.
  • the anti-AAV monospecific antibodies or anti-AAV x anti-cell surface molecule multispecific antibodies of the present disclosure can be linked to or co-expressed with another functional molecule, e.g., another peptide or protein.
  • another functional molecule e.g., another peptide or protein.
  • an antibody or fragment thereof can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association, or otherwise) to one or more other molecular entities, such as another antibody or antibody fragment to produce a multispecific antibody with a second or additional binding specificity.
  • anti-AAV antibody or “anti-cell surface molecule antibody” herein is intended to include both monospecific anti-AAV antibodies or anti-cell surface antibodies as well as multispecific antibodies (e.g., bispecific antibodies) which may ⁇ ⁇ Attorney Docket No.250298.000954 comprise a first-binding domain that binds to a capsid of an AAV and a second binding domain that binds to a molecule on a cell surface.
  • the present disclosure includes bispecific antibodies wherein one domain of an immunoglobulin binds to a capsid of an AAV particle (e.g., a capsid comprising a wild-type and/or non-wild-type AAV capsid protein(s)), and the other domain of the immunoglobulin binds to a molecule of a cell surface.
  • the AAV- binding domain can comprise any of the CDR, HCVR/LCVR, or HC/LC amino acid sequences as set forth in Table 1 herein.
  • the cell surface molecule-binding domain can comprise any of various CDR, HCVR/LCVR, or HC/LC amino acid sequences that bind any of various cell-surface molecules disclosed herein, or otherwise within the knowledge of one skilled in the art.
  • the cell surface molecule-binding domain can comprise any of CDR, HCVR/LCVR, or HC/LC amino acid sequences which bind to Asialoglycoprotein Receptor 1 (ASGR1) as set forth in Table 5 herein.
  • Nucleic acid molecules encoding any of the CDR, HCVR/LCVR, or HC/LC amino acid sequences of the anti-AAV/anti-cell surface molecule multispecific antigen-binding molecules disclosed herein may include, for example, nucleic acid molecules comprising the polynucleotide sequences as set forth in Table 2 herein, as well as, e.g., nucleic acid molecules comprising the polynucleotide sequences as set forth in Table 6 herein.
  • MBMs of the present disclosure may comprise two or more (e.g., three, four) antigen-binding domains.
  • the two or more (e.g., three, four) antigen-binding domains may independently be in a Fab or an scFv format.
  • Single chain Fv or “scFv” antibody fragments comprise the VH and VL domains of an antibody in a single polypeptide chain, are capable of being expressed as a single chain polypeptide and retain the specificity of the intact antibodies from which they are derived.
  • an scFv polypeptide may further comprise a polypeptide linker between the VH and VL domain that enables the scFv to form the desired structure for target binding.
  • linkers suitable for connecting the V H and V L chains of an scFV are the linkers are described herein.
  • an scFv may have the V L and V H variable regions in either order, e.g., with respect to the N-terminal and C-terminal ends of the polypeptide, the scFv may comprise VL-linker-VH or may comprise VH-linker-VL.
  • an scFv comprising V L -linker-V H which may be used in accordance with the disclosure may comprise the amino acid sequence of DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID ⁇ ⁇ Attorney Docket No.250298.000954 SWGQGALVTVSS (SEQ ID NO: 1358), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity
  • an scFv comprising VL-linker-VH which may be used in accordance with the disclosure may comprise the amino acid sequence of DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGCGTKVEIKGGGGSGGGGSGGG GSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKCLEWVAVI WHDGSDKYYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGL GTLVTVSS (SEQ ID NO: 1355), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • an scFv comprising VH-linker-VL which may be used in accordance with the disclosure may comprise the amino acid sequence of ⁇ QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKCLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS GGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQ KPGKAPNLLIYKASNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFG CGTKVEIK (SEQ ID NO: 1350), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the scFv can comprise VL and VH sequences from any suitable species, such as murine, human, or humanized VH and VL sequences.
  • the VL and VH-encoding DNA fragments are operably linked to another fragment encoding a linker, e.g., encoding any of the linkers described herein, such that the V L and V H sequences can be expressed as a contiguous single-chain protein, with the VL and VH regions joined by the flexible linker (see, e.g., Bird et al., 1988, Science 242:423- 426; Huston et ai, 1988, Proc. Natl.
  • the MBMs of the disclosure may comprise at least one Fab domain.
  • Fab domains were traditionally produced from by proteolytic cleavage of immunoglobulin molecules using enzymes such as papain.
  • the Fab domains are recombinantly expressed as part of a larger molecule.
  • the Fab domains can comprise constant domain and variable region sequences from any suitable species, and thus can be murine, chimeric, human, or humanized.
  • Fab domains typically comprise a CH1 domain attached to a VH domain which pairs with a CL domain attached to a V L domain.
  • VH domain is paired with the VL domain to constitute the Fv region
  • CH1 domain is paired with the CL domain to further stabilize the binding module.
  • a disulfide bond between ⁇ ⁇ Attorney Docket No.250298.000954 the two constant domains can further stabilize the Fab domain.
  • correct association between the two ⁇ ⁇ Attorney Docket No.250298.000954 polypeptides of a Fab is promoted by exchanging the V L and V H domains of the Fab for each other or exchanging the CH1 and CL domains for each other, e.g., as described in WO 2009/080251.
  • Correct Fab pairing can also be promoted by introducing one or more amino acid modifications in the CH1 domain and one or more amino acid modifications in the CL domain of the Fab and/or one or more amino acid modifications in the VH domain and one or more amino acid modifications in the V L domain.
  • the amino acids that are modified are typically part of the VH:VL and CH1:CL interface such that the Fab components preferentially pair with each other rather than with components of other Fabs.
  • the one or more amino acid modifications are limited to the conserved framework residues of the variable (VH, VL) and constant (CH1, CL) domains as indicated by the Kabat numbering of residues.
  • VH, VL variable
  • CH1, CL constant domains
  • the modifications introduced in the V H and CH1 and/or V L and CL domains are complementary to each other.
  • Complementarity at the heavy and light chain interface can be achieved on the basis of steric and hydrophobic contacts, electrostatic/charge interactions, or a combination of the variety of interactions.
  • the complementarity between protein surfaces is broadly described in the literature in terms of lock and key fit, knob into hole, protrusion and cavity, donor and acceptor, etc., all implying the nature of structural and chemical match between the two interacting surfaces.
  • the one or more introduced modifications introduce a new hydrogen bond across the interface of the Fab components.
  • the one or more introduced modifications introduce a new salt bridge across the interface of the Fab components. Exemplary substitutions are described in WO 2014/150973 and WO 2014/082179, the contents of which are hereby incorporated by reference.
  • the Fab domain comprises a 192E substitution in the CH1 domain and 114A and 137K substitutions in the CL domain, which introduces a salt-bridge between the CFM and CL domains (see, e.g., Golay et al., 2016, J Immunol 196:3199-211).
  • the Fab domain comprises a 143Q and 188V substitutions in the CH1 domain and 113T and 176V substitutions in the CL domain, which serves to swap hydrophobic and polar regions of contact between the CH1 and CL domain (see, e.g., Golay et al., 2016, J Immunol 196:3199-211).
  • the Fab domain can comprise modifications in some or all of the VH, CH1, VL, CL domains to introduce orthogonal Fab interfaces which promote correct assembly of Fab domains (Lewis et al., 2014 Nature Biotechnology 32:191-198).
  • 39K, 62E modifications are introduced in the VH domain
  • H172A, F174G modifications are introduced in the CH1 domain
  • 1 R, 38D, (36F) modifications are introduced in the VL domain
  • L135Y, S176W modifications are introduced in the CL domain.
  • a 39Y modification is introduced in the V H domain and a 38R modification is introduced in the VL domain.
  • Fab domains can also be modified to replace the native CH1:CL disulfide bond with an engineered disulfide bond, thereby increasing the efficiency of Fab component pairing.
  • an engineered disulfide bond can be introduced by introducing a 126C in the CH1 domain and a 121 C in the CL domain (see, e.g., Mazor et al., 2015, Mabs 7:377-89).
  • Fab domains can also be modified by replacing the CH1 domain and CL domain with alternative domains that promote correct assembly.
  • V L of common light chain (also referred to as a universal light chain) can be used for each Fab VL region of a MBM of the disclosure.
  • employing a common light chain as described herein reduces the number of inappropriate species of MBMs as compared to employing original cognate VLs.
  • the VL domains of the MBMs are identified from monospecific antibodies comprising a common light chain.
  • the V H regions of the MBMs comprise human heavy chain variable gene segments that are rearranged in vivo within mouse B cells that have been previously engineered to express a limited human light chain repertoire, or a single human light chain, cognate with human heavy chains and, in response to exposure with an antigen of interest, generate an antibody repertoire containing a plurality of human V H s that are cognate with one or one of two possible human VLs, wherein the antibody repertoire specific for the antigen of interest.
  • Common light chains are those derived from a rearranged human VK1-39JK5 sequence or a rearranged human VK3-20JK1 sequence, and include somatically mutated (e.g., affinity matured) versions. See, for example, U.S. Patent No.10,412,940.
  • a universal light chain which may be used in accordance with the disclosure may comprise the amino acid sequence of DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ ⁇ ⁇
  • Attorney Docket No.250298.000954 LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20) which may be encoded by the nucleic acid sequence of GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCA CCATCACTTGCCGGGCAAGTCAGCATTAGCAGCTATTTAAATTGGTATC
  • MBMs of the present disclosure may comprise an antigen- binding domain which is in a single-domain antibody (sdAb) format.
  • sdAb describes a single antigen-binding domain capable of binding to a cognate antigen.
  • sdAbs are often derived from heavy-chain only antibodies, however they also include single VH domains capable of binding to their cognate antigen in the absence of an associated light chain.
  • Heavy-chain only antibodies lack both light chains and a functional CH1 domain and thus rely exclusively on a heavy chain variable domain for antigen binding.
  • transgenic mammals e.g., mice
  • Such transgenic mammals include, for example, transgenic animals described in U.S. Patent Publications 2015/0289489 A1, 2023/0270086 A1, and 2023/0062964 A1, and 2020/0267951 A1, each of which is incorporated herein by reference.
  • a sdAb is generated by immunizing an animal that produces heavy-chain only antibodies, including a natural producer (e.g., camelids, sharks) or an engineered non-human mammal (e.g., a transgenic mouse), to obtain heavy-chain only antibodies.
  • a natural producer e.g., camelids, sharks
  • an engineered non-human mammal e.g., a transgenic mouse
  • Such antibodies may be screened to identify those having desirable properties (e.g., target affinity).
  • the variable region of the antibody heavy chain is cloned to construct a single domain antibody consisting of only one heavy chain variable region.
  • sdAbs can also be obtained by immunizing animals that generate traditional antibodies (e.g., rabbits) followed by screening for VHs having high binding affinity in the absence of their cognate light chain (see, e.g., Shinozaki et al., 2017, Scientific Reports, 7(1):5794).
  • sdAbs can be humanized by replacing natural (e.g., camelid) framework sequences with human sequences (see, e.g., Vincke, 2009, The Journal of Biological Chemistry, 285(5):3273-3284; Murakami et al., 2022, Antibodies, 11(1):10; and U.S.
  • a disulfide bond is introduced within a VHH to increase stability (see, e.g., Hagihara et al., 2007, The Journal of Biological Chemistry, 282(50):36489–36495).
  • the anti-AAV monospecific antibodies or anti-AAV x anti-cell surface molecule multispecific (e.g., bispecific) antibodies disclosed herein can comprise one or more amino acid substitutions, insertions, and/or deletions in the framework and/or CDR regions of the heavy chain variable domains as compared to the corresponding germline sequences from which the antibodies were derived.
  • antibodies, and antigen-binding fragments thereof which are derived from any of the amino acid sequences disclosed herein, wherein one or more amino acids within one or more framework and/or CDR regions are mutated to the corresponding residue(s) of the germline sequence from which the antibody was derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are referred to herein collectively as “germline mutations”), and having weak or no detectable binding to a AAV capsid antigen or a cell surface molecule antigen.
  • the antibodies and antigen-binding molecules of the present disclosure may comprise one or more amino acid substitutions, insertions, and/or deletions in the framework and/or CDR regions of the heavy and light chain variable domains as compared to the corresponding germline sequences from which the individual antigen-binding domains were derived.
  • Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available from, for example, public antibody sequence databases.
  • the antigen-binding molecules of the present disclosure may ⁇ ⁇ Attorney Docket No.250298.000954 comprise antigen-binding domains which are derived from any of the exemplary amino acid sequences disclosed herein, wherein one or more amino acids within one or more framework and/or CDR regions are mutated to the corresponding residue(s) of the germline sequence from which the antibody was derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are referred to herein collectively as “germline mutations”).
  • a person of ordinary skill in the art can easily produce numerous antibodies and antigen-binding fragments which comprise one or more individual germline mutations or combinations thereof.
  • all of the framework and/or CDR residues within the VH and/or VL domains are mutated back to the residues found in the original germline sequence from which the antigen-binding domain was originally derived.
  • only certain residues are mutated back to the original germline sequence, e.g., only the mutated residues found within the first 8 amino acids of FR1 or within the last 8 amino acids of FR4, or only the mutated residues found within CDR1, CDR2, or CDR3.
  • a light chain sequence within an antibody or antigen-binding molecule disclosed herein may include one or more amino acid mutations designed to reduce steric hindrance between the light chain and its paired heavy chain. In some embodiments, such amino acid mutations may enhance the stability and/or yield of the antibody or antigen-binding molecule.
  • a light chain amino acid sequence such as, but not limited to, DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813) may comprise, one or more of a K50A or a A51K mutation.
  • a light chain sequence described herein comprises a K50A mutation. In some embodiments, a light chain sequence described herein comprises a A51K. In some embodiments, a light chain sequence described herein comprises a K50A mutation and a A51K mutation.
  • the antigen-binding domains may contain any combination of two or more germline mutations within the framework and/or CDR regions, e.g., wherein certain individual residues are mutated to the corresponding residue of a particular germline ⁇ ⁇ Attorney Docket No.250298.000954 sequence while certain other residues that differ from the original germline sequence are maintained or are mutated to the corresponding residue of a different germline sequence.
  • antigen-binding domains that contain one or more germline mutations can be easily tested for one or more desired property such as, improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as the case may be), reduced immunogenicity, etc.
  • Bispecific antigen-binding molecules comprising one or more antigen-binding domains obtained in this general manner are encompassed within the present disclosure.
  • the present disclosure provides MBM in which two or more components of an ABD (e.g., a VH and a VL of an scFv), two or more ABDs (e.g., an scFv and a Fab, or a Fab and a Fab), or an ABD and a non-ABD component (e.g., an Fc region) are connected to one another by a peptide linker.
  • an ABD e.g., a VH and a VL of an scFv
  • ABDs e.g., an scFv and a Fab, or a Fab and a Fab
  • a non-ABD component e.g., an Fc region
  • a peptide linker can range from 2 amino acids to 60 or more amino acids, and in certain aspects a peptide linker ranges from 3 amino acids to 50 amino acids, from 4 to 30 amino acids, from 5 to 25 amino acids, from 10 to 25 amino acids, 10 amino acids to 60 amino acids, from 12 amino acids to 20 amino acids, from 20 amino acids to 50 amino acids, or from 25 amino acids to 35 amino acids in length.
  • a peptide linker e.g., a peptide linker separating an scFv domain and a heavy chain constant region, is at least 5 amino acids, at least 6 amino acids, or at least 7 amino acids in length and optionally is up to 30 amino acids, up to 40 amino acids, up to 50 amino acids, or up to 60 amino acids in length.
  • the linker ranges from 5 amino acids to 50 amino acids in length, e.g., ranges from 5 to 50, from 5 to 45, from 5 to 40, from 5 to 35, from 5 to 30, from 5 to 25, or from 5 to 20 amino acids in length.
  • the linker ranges from 6 amino acids to 50 amino acids in length, e.g., ranges from 6 to 50, from 6 to 45, from 6 to 40, from 6 to 35, from 6 to 30, from 6 to 25, or from 6 to 20 amino acids in length.
  • the linker ranges from 7 amino acids to 50 amino acids in length, e.g., ranges from 7 to 50, from 7 to 45, from 7 to 40, from 7 to 35, from 7 to 30, from 7 to 25, or from 7 to 20 amino acids in length.
  • Charged (e.g., charged hydrophilic linkers) and/or flexible linkers are particularly preferred.
  • Examples of flexible linkers that can be used in the MBMs of the disclosure include those disclosed by Chen et ai, 2013, Adv Drug Deliv Rev.65(10): 1357-1369 and Klein et al., 2014, Protein Engineering, Design & Selection 27(10): 325-330.
  • Particularly useful flexible linkers are or comprise repeats of glycines and serines, e.g., a monomer or multimer ⁇ ⁇ Attorney Docket No.250298.000954 of G n S (SEQ ID NO: 239) or SG n (SEQ ID NO: 240), wherein n is an integer from 1 to 10, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the linker is or comprises a monomer or multimer of repeat of G4S (SEQ ID NO: 242), e.g., (GGGGS)n (SEQ ID NO: 241), wherein n is an integer from 1 to 10, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the linker is or comprises, e.g., (GGGGS)3 (SEQ ID NO: 1115).
  • the linker is or comprises, e.g., (GGGGS)4 (SEQ ID NO: 1161).
  • Polyglycine linkers can suitably be used in the MBMs of the disclosure.
  • the peptide linker e.g., a peptide linker separating an scFv domain and a heavy chain such as the scFv domain of ABD1 and the heavy chain variable region of ABD2, comprises two consecutive glycines (2Gly), three consecutive glycines (3Gly), four consecutive glycines (4Gly) (SEQ ID NO: 243), five consecutive glycines (5Gly) (SEQ ID NO: 244), six consecutive glycines (6Gly) (SEQ ID NO: 245), seven consecutive glycines (7Gly) (SEQ ID NO: 246), eight consecutive glycines (8Gly) (SEQ ID NO: 247), or nine consecutive glycines (9Gly) (SEQ ID NO: 248).
  • the linker e.g., a peptide linker separating an scFv domain and a heavy chain constant region, is composed of both G4S (SEQ ID NO: 242) or a multimer thereof and one or more additional glycines, e.g., 2Gly, 3Gly, or 4Gly (SEQ ID NO: 243).
  • additional glycines e.g., 2Gly, 3Gly, or 4Gly (SEQ ID NO: 243.
  • Examples of such linkers include G 4 S (SEQ ID NO: 242), GG, 4xG 4 S GG (SEQ ID NO: 249), and 7xG4S GG (SEQ ID NO: 250).
  • Non-limiting examples of linker sequences are set forth in Table L below.
  • a MBM of the disclosure may comprise one or more linkers of Table L. Table L. Linker Sequences ⁇ ⁇ Attorney Docket No.250298.000954 ⁇ ⁇ Attorney Docket No.250298.000954 Constant Regions [00451]
  • MBMs of the disclosure comprise constant regions (e.g., CH1, hinge, CH2, CH3, CL) derived from any suitable class of antibody.
  • the constant regions are derived from a human antibody.
  • the constant regions can be derived from any suitable class of antibody, including IgA (including subclasses lgA1 and lgA2), IgD, IgE, IgG (including subclasses lgG1, lgG2, lgG3, and lgG4), and IgM.
  • the Fc domain is derived from lgG1, lgG2, lgG3, or lgG4.
  • the constant region is derived from lgG1.
  • the Fc domain is derived from lgG4.
  • a heavy chain constant region of a human IgG1 which may be used in accordance with the disclosure may comprise the amino acid sequence of ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSPGK (SEQ ID NO: 259) which may be encoded by the nucleic acid sequence
  • a heavy chain constant region of a human IgG1 which may be used in accordance with the disclosure may comprise the amino acid sequence of ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSPGK (SEQ ID NO: 1164) which may be encoded by the nucleic acid
  • a heavy chain constant region of a human IgG1 which may be used in accordance with the disclosure may comprise the amino acid sequence of ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKN QVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGN VFSCSVMHEALHNRFTQKSLSPGK (SEQ ID NO: 1144) which may be encoded by the nucleic acid sequence of
  • a heavy chain constant region of a human IgG1 which may be used in accordance with the disclosure may comprise the amino acid sequence of ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTSKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRD ELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSPGK (SEQ ID NO: 1178) which may be encoded by
  • a heavy chain constant region of a human IgG4, hIgG4us which may be used in accordance with the disclosure may comprise the amino acid sequence of ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPGGGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSRLTVDKSRWQEGNVFS CSVMHEALHNRFTQKSLSLSPGK (SEQ ID NO: 1364), or
  • a heavy chain constant region of a human IgG4, hIgG4us which may be used in accordance with the disclosure may comprise the amino acid sequence of ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPGGGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFS CSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 1377),
  • a heavy chain Fc region of a human IgG4, hIgG4us which may be used in accordance with the disclosure may comprise the amino acid sequence of ⁇ ⁇ Attorney Docket No.250298.000954 ESKYGPPCPPCPAPGGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYV DGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 1397), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • a heavy chain Fc region of a human IgG4, hIgG4us which may be used in accordance with the disclosure may comprise the amino acid sequence of ESKYGPPCPPCPAPGGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYV DGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK GQPREPQVYTLPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLVSRLTVDKSRWQEGNVFSCSVMHEALHNRFTQKSLSLSPGK (SEQ ID NO: 1365), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • a heavy chain Fc region of a human IgG4, hIgG4us which may be used in accordance with the disclosure may comprise the amino acid sequence of ⁇ ESKYGPPCPPCPAPGGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYV DGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK GQPREPQVYTLPPSQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 1345), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • a heavy chain Fc region of a human IgG4, hIgG4us which may be used in accordance with the disclosure may comprise the amino acid sequence of ⁇ ESKYGPPCPPCPAPGGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYV DGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNRFTQKSLSLSPGK (SEQ ID NO: 1511), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the MBMs of the disclosure can also comprise hinge regions, e.g., connecting an ABD module to an Fc region.
  • the hinge region can be a native or a modified hinge region. Hinge regions are typically found at the N-termini of Fc regions.
  • a native hinge region is the hinge region that would normally be found between Fab and Fc domains in a naturally occurring antibody.
  • a modified hinge region is any hinge that ⁇ ⁇ Attorney Docket No.250298.000954 differs in length and/or composition from the native hinge region.
  • Such hinges can include hinge regions from other species, such as human, mouse, rat, rabbit, shark, pig, hamster, camel, llama, or goat hinge regions.
  • Other modified hinge regions may comprise a complete hinge region derived from an antibody of a different class or subclass from that of the heavy chain Fc region.
  • the modified hinge region may comprise part of a natural hinge or a repeating unit in which each unit in the repeat is derived from a natural hinge region.
  • the natural hinge region may be altered by converting one or more cysteine or other residues into neutral residues, such as serine or alanine, or by converting suitably placed residues into cysteine residues.
  • modified hinge regions may be entirely synthetic and may be designed to possess desired properties such as length, cysteine composition, and flexibility.
  • a number of modified hinge regions have already been described for example, in U.S. Patent No.5,677,425, WO9915549, WO2005003170, WO2005003169, WO2005003170, WO9825971, and WO2005003171, and these are incorporated herein by reference.
  • the Fc region of one or both chains of the MBMs of the disclosure possess an intact hinge region, e.g., a hinge domain, at its N-terminus.
  • hinge domain refers to the sequence from about Glu216 or about Cys226 to about Pro230 of human lgG1 (Burton, 1985 Molec. Immunol.22:161-206), or the corresponding sequence in another antibody class or isotype. [00469] In various embodiments, positions 233-236 within a hinge domain may be G, G, G, and unoccupied; G, G, unoccupied, and unoccupied; G, unoccupied, unoccupied, and unoccupied; or all unoccupied, with positions numbered by EU numbering.
  • the MBMs of the disclosure comprise a modified hinge domain that reduces binding affinity for an Fc ⁇ receptor relative to a wild-type hinge domain of the same isotype (e.g., human lgG1 or human lgG4).
  • a wild-type hinge domain of the same isotype e.g., human lgG1 or human lgG4.
  • the Fc region of one or both heavy chains of the MBMs of disclosure possesses an intact hinge domain at its N-terminus.
  • both the Fc region and the hinge region of an MBM of the disclosure are derived from lgG4 and the hinge region comprises the modified sequence CPPC (SEQ ID NO: 251).
  • the core hinge region of human lgG4 contains the sequence CPSC (SEQ ID NO: 252) compared to lgG1 that contains the sequence CPPC (SEQ ID NO: 251).
  • the serine residue which may be present in the lgG4 sequence can lead to increased flexibility in this region, and therefore a proportion of molecules form disulfide ⁇ ⁇ Attorney Docket No.250298.000954 bonds within the same protein chain (an intrachain disulfide) rather than bridging to the other heavy chain in the IgG molecule to form the interchain disulfide (Angel et ai, 1993, Mol Immunol 30(1 ): 105-108).
  • the hinge region can be a chimeric hinge region.
  • a chimeric hinge may comprise an “upper hinge” sequence, derived from a human lgG1, a human lgG2, or a human lgG4 hinge region, combined with a “lower hinge” sequence, derived from a human lgG1, a human lgG2, or a human lgG4 hinge region.
  • a chimeric hinge region comprises the amino acid sequence EPKSCDKTHTCPPCPAPPVA (SEQ ID NO: 253) (see, e.g., SEQ ID NO: 8 of WO2014/121087, which is incorporated by reference in its entirety herein), ESKYGPPCPPCPAPPVA (SEQ ID NO: 254) (see, e.g., SEQ ID NO: 9 of WO2014/121087), or ESKYGPPCPPCPAPGGG (SEQ ID NO: 1371) (see, e.g., SEQ ID NO: 9 of ⁇ WO2021/226444, which is incorporated by reference in its entirety herein). [00477] .
  • Such chimeric hinge sequences can be suitably linked to an lgG4 CH2 region (for example by incorporation into an lgG4 Fc domain, for example a human or murine Fc domain, which can be further modified in the CH2 and/or CH3 domain to reduce effector function.
  • the hinge region can be modified to reduce effector function, for example as described in WO2016161010, which is incorporated by reference in its entirety herein.
  • the positions 233-236 of the modified hinge region are G, G, G, and unoccupied; G, G, unoccupied, and unoccupied; G, unoccupied, unoccupied, and unoccupied; or all unoccupied, with positions numbered by EU numbering (as shown in FIG.1 of WO2016161010A2).
  • These segments can be represented as GGG-, GG-, G — or – with representing an unoccupied position.
  • Position 236 is unoccupied in canonical human lgG2 but is occupied by in other canonical human IgG isotypes.
  • Positions 233-235 are occupied by residues other than G in all four human isotypes (as shown in FIG.1 of WO2016161010A2).
  • positions 233-236 can be combined with position 228 being occupied by P.
  • Position 228 is naturally occupied by P in human lgG1 and lgG2 but is occupied by S in human lgG4 and R in human lgG3.
  • An S228P mutation in an lgG4 antibody is advantageous in stabilizing an lgG4 antibody and reducing exchange of heavy chain light chain pairs between exogenous and endogenous antibodies.
  • positions 226-229 are occupied by C, P, P, and C respectively.
  • Exemplary hinge regions have residues 226-236, sometimes referred to as middle ⁇ ⁇ Attorney Docket No.250298.000954 (or core) and lower hinge, occupied by the modified hinge sequences designated GGG-(233- 236), GG-(233-236), G— (233-236), and no G(233-236).
  • the hinge domain amino acid sequence comprises CPPCPAPGGG-GPSVF (SEQ ID NO: 255) (see, e.g., SEQ ID NO:1 of WO2016161010A2), CPPCPAPGG-GPSVF (SEQ ID NO: 256) (see, e.g., SEQ ID NO:2 of WO2016161010A2), CPPCPAPG— GPSVF (SEQ ID NO: 257) (see, e.g., SEQ ID NO:3 of WO2016161010A2), or CPPCPAP — GPSVF (SEQ ID NO: 258) (see, e.g., SEQ ID NO:4 of WO2016161010A2).
  • SEQ ID NO: 255 see, e.g., SEQ ID NO:1 of WO2016161010A2
  • CPPCPAPGG-GPSVF SEQ ID NO: 256
  • CPPCPAPG— GPSVF SEQ ID NO: 257
  • the modified hinge regions described above can be incorporated into a heavy chain constant region, which typically include CH2 and CH3 domains, and which may have an additional hinge segment (e.g., an upper hinge) flanking the designated region.
  • additional constant region segments present are typically of the same isotype, preferably a human isotype, although can be hybrids of different isotypes.
  • the isotype of such additional human constant regions segments is preferably human lgG4 but can also be human lgG1, lgG2, or lgG3 or hybrids thereof in which domains are of different isotypes. Exemplary sequences of human lgG1, lgG2, and lgG4 are shown in FIGS.2-4 of WO2016161010A2.
  • the modified hinge sequences can be linked to an lgG4 CH2 region (for example by incorporation into an lgG4 Fc domain, for example a human or murine Fc domain, which can be further modified in the CH2 and/or CH3 domain to reduce effector function).
  • Fc Domains [00485]
  • the MBMs of the disclosure can include an Fc region derived from any suitable species. In one embodiment the Fc region is derived from a human Fc domain.
  • MBMs disclosed herein can have, e.g., fully human variable regions but can have mouse constant regions (e.g., a mouse IgG1 Fc or a mouse IgG2 Fc (a or b isotype)) or human constant regions (e.g., a human IgG1 Fc or a human IgG4 Fc).
  • mouse constant regions e.g., a mouse IgG1 Fc or a mouse IgG2 Fc (a or b isotype)
  • human constant regions e.g., a human IgG1 Fc or a human IgG4 Fc
  • a MBM having a particular Fc isotype can be converted to an antibody with a different Fc isotype (e.g., an antibody with a mouse IgG1 Fc can be converted to an antibody with a human IgG4, etc.), but in any event, the variable domains (including the CDRs) will remain the same, and the binding properties to antigen are expected to be identical or substantially similar regardless of the nature of the constant domain.
  • the Fc domain can be derived from any suitable class of antibody, including IgA (including subclasses lgA1 and lgA2), IgD, IgE, IgG (including subclasses lgG1, lgG2, lgG3, and lgG4), and IgM.
  • the Fc domain is derived from lgG1, lgG2, lgG3, or lgG4.
  • the Fc domain is derived from lgG1.
  • the Fc domain is derived from lgG4.
  • Fc domains derived from IgG4 are [00489] ESKYGPPCPPCPAPGGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEV QFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE KTISKAKGQPREPQVYTLPPSQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 1345), and ESKYGPPCPPCPAPGGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYV DGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEY
  • the two Fc domains within the Fc region can be the same or different from one another.
  • the Fc domains are typically identical, but for the purpose of producing multispecific binding molecules of the disclosure, the Fc domains might advantageously be different to allow for heterodimerization.
  • the heavy chain Fc domain of IgA, IgD, and IgG is composed of two heavy chain constant domains (CH2 and CH3) and that of IgE and IgM is composed of three heavy chain constant domains (CH2, CH3, and CH4). These dimerize to create an Fc region.
  • the Fc region, and/or the Fc domains within it can comprise heavy chain constant domains from one or more different classes of antibody, for example one, two, or three different classes.
  • the Fc region comprises CH2 and CH3 domains derived from lgG1.
  • the Fc region comprises CH2 and CH3 domains derived from lgG2.
  • the Fc region comprises CH2 and CH3 domains derived from lgG3.
  • the Fc region comprises CH2 and CH3 domains derived from lgG4.
  • the Fc region comprises a CH4 domain from IgM.
  • the IgM CH4 domain is typically located at the C-terminus of the CH3 domain.
  • the Fc region comprises CH2 and CH3 domains derived from IgG and a CH4 domain derived from IgM.
  • the heavy chain constant domains for use in producing an Fc region for the MBMs of the present disclosure may include variants of the naturally ⁇ ⁇ Attorney Docket No.250298.000954 occurring constant domains described above. Such variants may comprise one or more amino acid variations compared to wild type constant domains.
  • the Fc region of the present disclosure comprises at least one constant domain that varies in sequence from the wild-type constant domain.
  • the variant constant domains may be longer or shorter than the wild-type constant domain.
  • the variant constant domains are at least 60% identical or similar to a wild-type constant domain.
  • the variant constant domains are at least 70% identical or similar.
  • the variant constant domains are at least 80% identical or similar.
  • the variant constant domains are at least 90% identical or similar.
  • the variant constant domains are at least 95% identical or similar.
  • IgM occurs as a pentamer when it has incorporated a J-chain, or as a hexamer when it lacks a J-chain.
  • IgA occurs as monomer and dimer forms.
  • the heavy chains of IgM and IgA possess an 18 amino acid extension to the C-terminal constant domain, known as a tailpiece.
  • the tailpiece includes a cysteine residue that forms a disulfide bond between heavy chains in the polymer and is believed to have an important role in polymerization.
  • the tailpiece also contains a glycosylation site.
  • the MBMs of the present disclosure do not comprise a tailpiece.
  • the Fc domains that are incorporated into the MBMs of the present disclosure may comprise one or more modifications that alter the functional properties of the proteins, for example, binding to Fc-receptors such as FcRn or leukocyte receptors, binding to complement, modified disulfide bond architecture, or altered glycosylation patterns.
  • the Fc domains can also be altered to include modifications that improve manufacturability of asymmetric MBMs, for example by allowing heterodimerization, which is the preferential pairing of non-identical Fc domains over identical Fc domains. Heterodimerization permits the production of MBMs in which different A BSs are connected to one another by an Fc region containing Fc domains that differ in sequence.
  • Fc domain sequences are set forth in Table F-1 below.
  • a MBM of the disclosure may comprise one or more Fc domain sequences of Table F-1. ⁇ ⁇ Attorney Docket No.250298.000954 Table F-1.
  • an Fc domain comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% sequence identity to any one of the sequences disclosed in Table F-1.
  • the Fc domain comprises one or more amino acid substitutions that reduces binding to an Fc receptor and/or effector function.
  • the Fc receptor is an Fc ⁇ receptor.
  • the Fc receptor is a human Fc receptor.
  • the Fc receptor is an activating Fc receptor.
  • the Fc receptor is an activating human Fey receptor, ⁇ ⁇ Attorney Docket No.250298.000954 more specifically human FcyRIIIa, FcyRI, or FcyRIla, most specifically human FcyRIIIa.
  • the effector function is one or more selected from the group of complement dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and cytokine secretion.
  • the effector function is ADCC.
  • the Fc region comprises an amino acid substitution at a position selected from the group of E233, L234, L235, N297, P331, and P329 (numberings according to Kabat EU index). In a more specific embodiment, the Fc region comprises an amino acid substitution at a position selected from the group of L234, L235, and P329 (numberings according to Kabat EU index). In some embodiments, the Fc region comprises the amino acid substitutions L234A and L235A (numberings according to Kabat EU index). In one such embodiment, the Fc region is an Igd Fc region, particularly a human Igd Fc region. In one embodiment, the Fc region comprises an amino acid substitution at position P329.
  • the amino acid substitution is P329A or P329G, particularly P329G (numberings according to Kabat EU index).
  • the Fc region comprises an amino acid substitution at position P329 and a further amino acid substitution at a position selected from E233, L234, L235, N297, and P331 (numberings according to Kabat EU index).
  • the further amino acid substitution is E233P, L234A, L235A, L235E, N297A, N297D, or P331S.
  • the Fc region comprises amino acid substitutions at positions P329, L234, and L235 (numberings according to Kabat EU index).
  • the Fc region comprises the amino acid mutations L234A, L235A, and P329G (“P329G LALA”, “PGLALA”, or “LALAPG”). [00509] Typically, the same one or more amino acid substitution is present in each of the two Fc domains of an Fc region.
  • each Fc domain of the Fc region comprises the amino acid substitutions L234A, L235A, and P329G (Kabat EU index numbering), i.e.
  • the leucine residue at position 234 is replaced with an alanine residue (L234A)
  • the leucine residue at position 235 is replaced with an alanine residue (L235A)
  • the proline residue at position 329 is replaced by a glycine residue (P329G) (numbering according to Kabat EU index).
  • the Fc domain is an lgG1 Fc domain, particularly a human lgG1 Fc domain.
  • the same one or more amino acid substitution is present in each of the two Fc domains of an Fc region.
  • each Fc domain of the Fc region comprises the amino acid substitutions L234A, L235A, and P329G (Kabat EU index numbering), i.e. in each of the first and the second Fc domains in the Fc region the leucine ⁇ ⁇ Attorney Docket No.250298.000954 residue at position 234 is replaced with an alanine residue (L234A), the leucine residue at position 235 is replaced with an alanine residue (L235A), and the proline residue at position 329 is replaced by a glycine residue (P329G) (numbering according to Kabat EU index).
  • the Fc domain is an lgG1 Fc domain, particularly a human lgG1 Fc domain.
  • the lgG1 Fc domain is a variant lgG1 comprising D265A, N297A mutations (EU numbering) to reduce effector function.
  • the Fc domain is an lgG4 Fc domain with reduced binding to Fc receptors.
  • Exemplary lgG4 Fc domains with reduced binding to Fc receptors may comprise an amino acid sequence selected from Table 4 below.
  • the Fc domain includes only the bolded portion of the sequences shown below. Table 4.
  • the lgG4 with reduced effector function comprises the bolded portion of the amino acid sequence of SEQ ID NO:31 of WO2014/121087, sometimes referred to herein as lgG4s or hlgG4s.
  • heterodimeric MBMs it is possible to incorporate a combination of the variant lgG4 Fc sequences set forth above, for example an Fc region comprising a combination of SEQ ID NO:30 of WO2014/121087 (or the bolded portion thereof) and SEQ ID NO:37 of WO2014/121087 (or the bolded portion thereof) or an Fc region comprising a combination of SEQ ID NO:31 of WO2014/121087 (or the bolded portion thereof) and SEQ ID NO:38 of WO2014/121087 (or the bolded portion thereof).
  • MBMs comprising Fc heterodimers, i.e., Fc regions comprising heterologous, non-identical Fc domains.
  • each Fc domain in the Fc heterodimer comprises a CH3 domain of an antibody.
  • the CH3 domains are derived from the constant region of an antibody of any isotype, class, or subclass, and preferably of IgG (lgG1, lgG2, lgG3, and lgG4) class, as ⁇ ⁇ Attorney Docket No.250298.000954 described in the preceding section.
  • the two heavy chains that associate to form an MBM of the disclosure will contain CH3 domains with modifications that favor heterodimeric association relative to unmodified chains.
  • said modification promoting the formation of Fc heterodimers is a so-called “knob-into-hole” or “knob-in-hole” (KiH) modification, comprising a “knob” modification in one of the Fc domains and a “hole” modification in the other Fc domain.
  • knob-into-hole technology is described e.g. in U.S. Patent No.5,731,168; US 7,695,936; Ridgway et al., 1996, Prot Eng 9:617-621, and Carter, 2001, Immunol Meth 248:7-15.
  • the method involves introducing a protuberance (“knob”) at the interface of a first polypeptide and a corresponding cavity (“hole”) in the interface of a second polypeptide, such that the protuberance can be positioned in the cavity so as to promote heterodimer formation and hinder homodimer formation.
  • Protuberances are constructed by replacing small amino acid side chains from the interface of the first polypeptide with larger side chains (e.g., tyrosine or tryptophan).
  • Compensatory cavities of identical or similar size to the protuberances are created in the interface of the second polypeptide by replacing large amino acid side chains with smaller ones (e.g., alanine or threonine).
  • an amino acid residue in the CH3 domain of the first subunit of the Fc domain is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the CH3 domain of the first subunit which is positionable in a cavity within the CH3 domain of the second subunit, and an amino acid residue in the CH3 domain of the second subunit of the Fc domain is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the CH3 domain of the second subunit within which the protuberance within the CH3 domain of the first subunit is positionable.
  • amino acid residue having a larger side chain volume is selected from arginine (R), phenylalanine (F), tyrosine (Y), and tryptophan (W).
  • amino acid residue having a smaller side chain volume is selected from alanine (A), serine (S), threonine (T), and valine (V).
  • the protuberance and cavity can be made by altering the nucleic acid encoding the polypeptides, e.g. by site-specific mutagenesis, or by peptide synthesis.
  • An exemplary substitution is Y470T.
  • the threonine residue at position 366 is replaced with a tryptophan residue (T366W), and in the Fc domain the tyrosine residue at position 407 is replaced with a valine residue (Y407V) and optionally the ⁇ ⁇ Attorney Docket No.250298.000954 threonine residue at position 366 is replaced with a serine residue (T366S) and the leucine residue at position 368 is replaced with an alanine residue (L368A) (numbering according to Kabat EU index).
  • the serine residue at position 354 is replaced with a cysteine residue (S354C) or the glutamic acid residue at position 356 is replaced with a cysteine residue (E356C) (particularly the serine residue at position 354 is replaced with a cysteine residue), and in the second Fc domain additionally the tyrosine residue at position 349 is replaced by a cysteine residue (Y349C) (numbering according to Kabat EU index).
  • the first Fc domain comprises the amino acid substitutions S354C and T366W
  • the second Fc domain comprises the amino acid substitutions Y349C, T366S, L368A, and Y407V (numbering according to Kabat EU index).
  • electrostatic steering e.g., as described in Gunasekaran et al., 2010, J Biol Chem 285(25): 19637-46) can be used to promote the association of the first and the second subunit of the Fc domain.
  • an Fc domain can be modified to allow a purification strategy that enables selections of Fc heterodimers.
  • one heavy chain comprises a modified Fc domain that abrogates its binding to Protein A, thus enabling a purification method that yields a heterodimeric protein. See, for example, U.S. Patent No. 8,586,713.
  • the MBMs comprise a first CH3 domain and a second Ig CH3 domain, wherein the first and second Ig CH3 domains differ from one another by at least one amino acid, and wherein at least one amino acid difference reduces binding of the MBM to Protein A as compared to a corresponding MBM lacking the amino acid difference.
  • the first CH3 domain binds Protein A and the second CH3 domain contains a mutation/modification that reduces or abolishes Protein A binding such as an H95R modification (by IMGT exon numbering; H435R by EU numbering).
  • the second CH3 may further comprise a Y96F modification (by IMGT; Y436F by EU). This class of modifications is referred to herein as “star” mutations.
  • anti-AAV antigen- binding molecules and anti-AAV x anti-cell surface molecule antigen-binding molecules are provided comprising an Fc domain comprising one or more mutations.
  • Fc modifications include, e.g., a modification at position 250 (e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., L/Y/F/W or T), 254 (e.g., S or T), and 256 (e.g., S/R/Q/E/D or T); or a modification at position 428 and/or 433 (e.g., H/L/R/S/P/Q or K) and/or 434 (e.g., H/F or Y); or a modification at position 250 and/or 428; or a modification at position 307 or 308 (e.g., 308F, V308F), and 434.
  • a modification at position 250 and/or 428 e.g., E or Q
  • the modification comprises a 428L ⁇ ⁇ Attorney Docket No.250298.000954 (e.g., M428L) and 434S (e.g., N434S) modification; a 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modification; a 433K (e.g., H433K) and a 434 (e.g., 434Y) modification; a 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modification; a 250Q and 428L modification (e.g., T250Q and M428L); and a 307 and/or 308 modification (e.g., 308F or 308P).
  • M428L e.g., M428L
  • 434S e.g., N434S
  • 428L, 259I e.g., V259I
  • the anti-AAV monospecific antibodies or anti-AAV x anti-cell surface molecule multispecific antibodies provided herein are human antibodies.
  • the term “human antibody”, as used herein, is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences.
  • the human antibodies of the disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example in the CDRs and, in particular, CDR3.
  • human antibody is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.
  • the antibodies of the disclosure may, in some embodiments, be recombinant human antibodies.
  • recombinant human antibody is intended to include all human antibodies that are prepared, expressed, created, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant, combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see e.g., Taylor et al. (1992) Nucl. Acids Res.20:6287-6295) or antibodies prepared, expressed, created, or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences.
  • Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the V H and V L regions of the recombinant antibodies are sequences that, while derived from and related to human germline V H and V L sequences, may not naturally exist within the human antibody germline repertoire in vivo. [00527] Human antibodies can exist in two forms that are associated with hinge heterogeneity.
  • an immunoglobulin molecule comprises a stable four chain construct of approximately 150-160 kDa in which the dimers are held together by an interchain heavy chain disulfide bond.
  • the dimers are not linked via inter- ⁇ ⁇ Attorney Docket No.250298.000954 chain disulfide bonds and a molecule of about 75-80 kDa is formed composed of a covalently coupled light and heavy chain (half-antibody). These forms have been extremely difficult to separate, even after affinity purification.
  • the frequency of appearance of the second form in various intact IgG isotypes is due to, but not limited to, structural differences associated with the hinge region isotype of the antibody.
  • a single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the appearance of the second form (Angal et al. (1993) Molecular Immunology 30:105) to levels typically observed using a human IgG1 hinge.
  • the instant disclosure encompasses antibodies having one or more mutations in the hinge, C H 2, or C H 3 region which may be desirable, for example, in production, to improve the yield of the desired antibody form.
  • the antibodies of the disclosure may be isolated antibodies.
  • An “isolated antibody,” as used herein, means an antibody that has been identified and separated and/or recovered from at least one component of its natural environment.
  • an antibody that has been separated or removed from at least one component of an organism, or from a tissue or cell in which the antibody naturally exists or is naturally produced is an “isolated antibody” for purposes of the present disclosure.
  • An isolated antibody also includes an antibody in situ within a recombinant cell. Isolated antibodies are antibodies that have been subjected to at least one purification or isolation step. According to certain embodiments, an isolated antibody may be substantially free of other cellular material and/or chemicals.
  • the anti-AAV monospecific antibodies or anti-AAV x anti-cell surface molecule multispecific antibodies disclosed herein may comprise one or more amino acid substitutions, insertions and/or deletions in the framework and/or CDR regions of the heavy and light chain variable domains as compared to the corresponding germline sequences from which the antibodies were derived. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available from, for example, public antibody sequence databases.
  • the present disclosure includes antibodies, and antigen-binding fragments thereof, which are derived from any of the amino acid sequences disclosed herein, wherein one or more amino acids within one or more framework and/or CDR regions are mutated to the corresponding residue(s) of the germline sequence from which the antibody was derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are referred to herein collectively as “germline mutations”).
  • a person of ordinary skill in the art can easily produce numerous antibodies and antigen-binding fragments which comprise one or more individual ⁇ ⁇ Attorney Docket No.250298.000954 germline mutations or combinations thereof.
  • all of the framework and/or CDR residues within the V H and/or V L domains are mutated back to the residues found in the original germline sequence from which the antibody was derived.
  • only certain residues are mutated back to the original germline sequence, e.g., only the mutated residues found within the first 8 amino acids of FR1 or within the last 8 amino acids of FR4, or only the mutated residues found within CDR1, CDR2, or CDR3.
  • one or more of the framework and/or CDR residue(s) are mutated to the corresponding residue(s) of a different germline sequence (i.e., a germline sequence that is different from the germline sequence from which the antibody was originally derived).
  • the antibodies of the present disclosure may contain any combination of two or more germline mutations within the framework and/or CDR regions, e.g., wherein certain individual residues are mutated to the corresponding residue of a particular germline sequence while certain other residues that differ from the original germline sequence are maintained or are mutated to the corresponding residue of a different germline sequence.
  • antibodies and antigen-binding fragments that contain one or more germline mutations can be easily tested for one or more desired property such as, improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as the case may be), reduced immunogenicity, etc.
  • Antibodies and antigen-binding fragments obtained in this general manner are encompassed within the present disclosure.
  • anti-AAV monospecific antibodies or anti-AAV x anti-cell surface molecule multispecific antibodies comprising variants of any of the HCVR, LCVR, and/or CDR amino acid sequences disclosed herein having one or more conservative substitutions.
  • the present disclosure includes anti-AAV monospecific antibodies or anti-AAV x anti-cell surface molecule multispecific antibodies having HCVR, LCVR, and/or CDR amino acid sequences with, e.g., 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, 3 or fewer, 2, or 1 conservative amino acid substitutions relative to any of the HCVR, LCVR, and/or CDR amino acid sequences set forth in Table 1 herein.
  • Multispecific Antigen-Binding Molecules Comprising Anti-AAV and Anti-Cell Surface Molecule Antigen-Binding Domains
  • the present disclosure provides antigen-binding molecules such as antibodies or fragments thereof, including multispecific antigen-binding molecules such as multispecific antibodies (e.g., bispecific antibodies) or fragments thereof, that can bind to a capsid of an adeno-associated virus (AAV) particle and/or a molecule on a cell surface (i.e., a cell surface molecule).
  • AAV adeno-associated virus
  • the antigen-binding molecule is a multispecific antigen- ⁇ ⁇ Attorney Docket No.250298.000954 binding molecule, e.g., a bispecific antigen-binding molecule.
  • the bispecific antigen-binding molecule is a bispecific antibody.
  • the multispecific antibodies (e.g., bispecific antibodies) provided herein comprise a first antigen-binding domain that binds to a capsid of an AAV particle, and a second antigen- binding domain that binds to a molecule on a cell surface (i.e., a cell surface molecule).
  • the capsid comprises a wild-type AAV capsid protein(s).
  • the capsid comprises a non-wild-type AAV capsid protein(s).
  • the first antigen-binding domain that binds to a capsid of an AAV particle comprises any of the HCVR amino acid sequences as set forth in Table 1.
  • the first antigen-binding domain that binds to a capsid of an AAV particle may also comprise any of the LCVR amino acid sequences as set forth in Table 1.
  • the first antigen- binding domain that binds to a capsid of a AAV particle comprises any of the HCVR/LCVR amino acid sequence pairs as set forth in Table 1.
  • anti- AAV/anti-cell surface multispecific antibodies e.g., bispecific antibodies
  • the first antigen-binding domain that binds to a capsid of an AAV particle comprises any of the heavy chain CDR1-CDR2-CDR3 amino acid sequences as set forth in Table 1, and/or any of the light chain CDR1-CDR2-CDR3 amino acid sequences as set forth in Table 1.
  • anti-AAV/anti-cell surface molecules wherein the first antigen-binding domain that binds to a capsid of an AAV particle comprises a heavy chain variable region (HCVR) having an amino acid sequence selected from SEQ ID NOs: 2, 22, 32, 42, 52, 1159, 62, 71, 81, 91, 101, 111, 131, 141, 151, 161, 171, 180, 190, 200, and 210, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • HCVR heavy chain variable region
  • anti-AAV/anti-cell surface molecule multispecific antibodies wherein the first antigen-binding domain that binds to a capsid of an AAV particle comprises a light chain variable region (LCVR) having an amino acid sequence selected from SEQ ID NOs: 10 and 119, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • LCVR light chain variable region
  • anti-AAV/anti-cell surface molecule multispecific antibodies wherein the first antigen-binding domain that binds to a capsid of an AAV particle comprises a HCVR and LCVR (HCVR/LCVR) amino acid sequence pair selected from SEQ ID Nos: 2/10 (e.g., REGN13876); 22/10 (e.g., REGN13877); 32/10 (e.g., REGN13878); 42/10 (e.g., REGN13879); 52 or 1159/10 or 1157 (e.g., REGN13880); 62/10 (e.g., REGN13881); 71/10 (e.g., REGN13882); 81/10 (e.g., REGN13883); 91/10 (e.g., REGN13884); 101/10 (e.g., REGN13885); 111/119 (e.g., REGN13070); 131/119 (e.g., REGN13071); 141/119 (e.g.
  • the first antigen-binding domain that binds to a capsid of an AAV particle comprises a heavy chain CDR1 (HCDR1) domain having an amino acid sequence selected from SEQ ID NOs: 4, 24, 34, 44, 54, 64, 73, 83, 93, 103, 113, 133, 143, 153, 163, 173, 182, 192, 202, and 212, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; a heavy chain CDR2 (HCDR2) domain having an amino acid sequence selected from SEQ ID NOs: 6, 26, 36, 46, 56, 56, 75, 85, 95, 105, 115, 135, 145, 155, 165, 145, 184, 194, 204, and 214, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%,
  • Certain non-limiting, exemplary anti-AAV/anti-cell surface molecule multispecific antibodies of the disclosure include a first antigen-binding domain that binds to a capsid of an AAV particle comprising HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 domains, respectively, having the amino acid sequences selected from: SEQ ID NOs: 4-6-8-12-14-16 (e.g. REGN13072); 153-155-157-121-123-125 (e.g., REGN13073); 163-165-167-121-123- 125 (e.g.
  • REGN13074 173-145-176-121-123-125 (e.g., REGN13075); 182-184-186-121- ⁇ ⁇ Attorney Docket No.250298.000954 123-125 (e.g. REGN13076); 192-194-196-121-123-125 (e.g., REGN13220); 202-204-206- 121-123-125 (e.g. REGN13221); and-212-214-216-121-123-125 (e.g. REGN13284).
  • anti-AAV/anti-cell surface multispecific antibodies wherein the second antigen-binding domain binds to a cell surface molecule (i.e., a molecule on the cell surface).
  • a cell surface molecule i.e., a molecule on the cell surface.
  • Such second antigen-binding domains may, therefore, bind to a protein(s) that is expressed on the surface of a cell in vitro or in vivo such that at least a portion of the protein is exposed to the extracellular side of the cell membrane and is accessible to the antigen-binding portion of the antigen-binding domain.
  • a cell surface molecule described herein may be a membrane protein, or a membrane-bound protein.
  • a membrane protein may be an integral membrane protein which can be a part of a cell membrane that can, e.g., penetrate the membrane (e.g., a transmembrane protein) and can span the whole way across the membrane, or can associate with one or the other side of a membrane (e.g., an integral monotopic protein).
  • the membrane protein may be permanently associated with the cell membrane.
  • the membrane protein may be transiently associated with the cell membrane.
  • a membrane protein described herein may be a multi-subunit protein.
  • Non-limiting examples cell surface molecules useful in the practice of the present disclosure include, e.g., Asialoglycoprotein Receptor 1 (ASGR1), ⁇ ⁇ , and Calcium Voltage-Gated Channel Auxiliary Subunit Gamma 1 (CACNG1).
  • ASGR1 Asialoglycoprotein Receptor 1
  • ASGR1 an exemplary molecule on the cell surface which may be bound by an anti- AAV/anti-cell surface multispecific antibody herein is Asialoglycoprotein Receptor 1 (ASGR1).
  • ASGR1 is not intended to be limiting.
  • anti-cell surface molecule antibodies may target any of various cell surface molecules known to a person of ordinary skill in the art.
  • an anti-AAV/anti-cell surface molecule multispecific antibody described herein may comprise a second antigen-binding domain that binds to cell surface molecule (e.g., ASGR1), wherein the second antigen-binding domain comprises any of the heavy chain variable region (HCVR) amino acid sequences as set forth in Table 5.
  • the second antigen-binding domain that binds to a cell surface molecule may also comprise any of the LCVR amino acid sequences as set forth in Table 5.
  • the second antigen-binding domain that binds to a cell surface molecule may comprise any of the HCVR/LCVR amino acid sequence pairs as set forth in Table 5.
  • the present disclosure also provides exemplary anti-AAV/anti-cell surface molecule multispecific antibodies, wherein the second antigen-binding domain that binds to a cell surface molecule comprises ⁇ ⁇ Attorney Docket No.250298.000954 any of the heavy chain CDR1-CDR2-CDR3 amino acid sequences as set forth in Table 5, and/or any of the light chain CDR1-CDR2-CDR3 amino acid sequences as set forth in Table 5.
  • nucleic acid molecules encoding any of the CDR, HCVR, LCVR, HC, or LC sequences of the anti-AAV/anti-cell surface molecule multispecific antigen-binding molecules disclosed herein, including nucleic acid molecules comprising the polynucleotide sequences as set forth in Table 2 herein, as well as nucleic acid molecules comprising the polynucleotide sequences as set forth in Table 6 herein, in any combination or arrangement thereof.
  • Recombinant expression vectors carrying the nucleic acids of the disclosure, and host cells into which such vectors have been introduced, are also encompassed by the disclosure, as are methods of producing the antibodies by culturing the host cells under conditions permitting production of the antibodies, and recovering the antibodies produced.
  • Table 5. Amino Acid Sequence Identifiers for Anti-ASGR1 Antibodies ⁇ Table 6.
  • anti-AAV/anti-cell surface molecule multispecific molecules wherein the second antigen-binding domain that binds molecule on the cell surface comprises a light chain variable region (LCVR) having an amino acid sequence of SEQ ID NOs: 10 or 1157, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • LCVR light chain variable region
  • anti-AAV/anti-cell surface molecule multispecific antibodies wherein the second antigen-binding domain that binds to a cell surface molecule comprises a HCVR and LCVR (HCVR/LCVR) amino acid sequence pair selected from SEQ ID NOs: 220 or 1239/10 or 1157 (e.g., REGN3954) and 230/10 (e.g., REGN3955).
  • HCVR/LCVR HCVR/LCVR
  • anti-AAV/anti-cell surface molecule multispecific antibodies wherein the second antigen-binding domain that binds to a molecule on the cell surface comprises a heavy chain CDR1 (HCDR1) domain having an amino acid sequence selected from SEQ ID NOs: 222 and 232, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; a heavy chain CDR2 (HCDR2) domain having an amino acid sequence selected from SEQ ID NOs: 224 and 234, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; a heavy chain CDR3 (HCDR3) domain having an amino acid sequence selected from SEQ ID NOs: 226 and 236, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; a light chain CDR1 (LCDR1) domain having an amino acid sequence of SEQ ID
  • the anti-ASGR1 antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 228 or 238, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 228 or 238.
  • the nucleotide sequence that encodes the anti- ASGR1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 228 or 238, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 228 or 238.
  • the nucleotide sequence that encodes the anti-ASGR1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 227 or 237, or a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the nucleotide ⁇ ⁇ Attorney Docket No.250298.000954 sequence of SEQ ID NOs: 227 or 237.
  • the anti-ASGR1 antibodies, or antigen-binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NOs: 228 or 238.
  • the nucleotide sequence that encodes the anti- ASGR1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 227 or 237.
  • the anti-ASGR1 antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NO: 20, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO: 20.
  • the nucleotide sequence that encodes the anti-ASGR1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NO: 20, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO: 20.
  • the nucleotide sequence that encodes the anti-ASGR1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NO: 19, or a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the nucleotide sequence of SEQ ID NO: 19.
  • the anti-ASGR1 antibodies, or antigen-binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NO: 20.
  • the nucleotide sequence that encodes the anti- ASGR1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NO: 19.
  • Certain non-limiting, exemplary anti-AAV/anti-cell surface molecule multispecific antibodies provided herein include a second antigen-binding domain that binds to a molecule on the cell surface comprising HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 domains, respectively, having the amino acid sequences selected from SEQ ID NOs: 222-224-226-12- 14-16 (e.g., REGN3954) and 232-234-236-12-14-16 (e.g., REGN3955).
  • the cell surface molecule which may be bound by an anti- AAV/anti-cell surface multispecific antibody herein is Transferrin Receptor (TfR).
  • an anti-AAV/anti-TfR multispecific antibody may comprise a second antigen-binding domain that binds to TfR, wherein the second antigen-binding domain comprises any of the heavy chain variable region (HCVR) amino acid sequences as set forth in Table 7.
  • the second antigen-binding domain that binds to TfR may also comprise any of the light chain variable region (LCVR) amino acid sequences as set forth in Table 7.
  • the second antigen-binding domain that binds to TfR may comprise any of the HCVR/LCVR amino acid sequence pairs as set forth in Table 7.
  • the present disclosure also ⁇ ⁇ Attorney Docket No.250298.000954 provides anti-AAV/anti-TfR multispecific antibodies, wherein the second antigen-binding domain that binds to TfR comprises any of the heavy chain CDR1-CDR2-CDR3 amino acid sequences as set forth in Table 7, and/or any of the light chain CDR1-CDR2-CDR3 amino acid sequences as set forth in Table 7.
  • nucleic acid molecules encoding any of the CDR, HCVR, LCVR, HC, or LC sequences of the anti-AAV/anti-TfR multispecific antigen- binding molecules disclosed herein, or variants thereof, including nucleic acid molecules comprising any of the polynucleotide sequences as set forth in Table 2 herein, as well as nucleic acid molecules comprising any of the polynucleotide sequences as set forth in Table 8 herein, in any combination thereof.
  • the nucleic acid molecules described herein may comprise one or more polynucleotide sequence(s) in Table 2 that encode a pair of HCVR/LCVR amino acid sequences, and one or more polynucleotide sequence(s) selected from Table 8 that encode a pair of HCVR/LCVR amino acid sequences.
  • the nucleic acid molecules described herein may comprise one or more polynucleotide sequence(s) selected from Table 2 that encode a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3), and one or more polynucleotide sequence(s) selected from Table 8 that encode a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3).
  • Table 7 Amino Acid Sequence Identifiers for Anti-TfR Antibodies ⁇ ⁇ Attorney Docket No.250298.000954 ⁇ ⁇ Attorney Docket No.250298.000954 Table 8.
  • HCVR Nucleic Acid Sequence Identifiers for Anti-TfR Antibodies 31874B HCVR (VH) Nucleotide Sequence GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAG ACTCTCCTGTGCAGCCTCTGGATTCGCCTTTAGCAGCTATGCCATGACCTGGGTCCGA CAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGTTATCAGTGGTACTGGTGGTAGT ⁇ Attorney Docket No.250298.000954 ACACGCTGTATCTACAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTG TGCGAAAGGGGGAGCAGCTCGTAGAATGGAATACTTCCAGTACTGGGGCCAGGGCAC CCTGGTCACCGTCTCCTCA (SEQ ID NO: 269) HCVR (VH) Amino Acid Sequence EVQLVESGGGLVQPGGSLRLSCAASGFAFSSYAMTWVRQAPGKG
  • anti-AAV/anti-TfR multispecific antibodies wherein the second antigen-binding domain that binds to TfR comprises a light chain variable region (LCVR) having an amino acid sequence selected from SEQ ID NOs: 276, 287, 298, 308, 309, 319, 320, 331, 332, 343, 344, 355, 356, 366, 377, 387, 398, 399, 409, 419, 430, 431, 441, 452, 462, 473, 474, 485, 496, 506, 516, 526, 536, 547, 557, 567, 577, 587, 598, 599, 609, 610, or 1127 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • LCVR light chain variable region
  • anti-AAV/anti-TfR multispecific antibodies wherein the second antigen-binding domain that binds to TfR comprises a HCVR and LCVR (HCVR/LCVR) amino acid sequence pair selected from SEQ ID NOs: 270 or 271/276 (e.g., REGN31874B), SEQ ID NOs: 281 or 282/287 (e.g., REGN31863B), SEQ ID NOs: 292 or 293/298 (e.g., REGN69348), SEQ ID NOs: 303/308 or 309 (e.g., REGN69340), SEQ ID NOs: 314/319 or 320 (e.g., REGN69331), SEQ ID NOs: 325 or 326/331 or 332 (e.g., REGN69332), SEQ ID NOs: 337 or 338/343 or 344 (e.g., REGN69326), SEQ ID NOs:
  • anti-AAV/anti-TfR multispecific antibodies wherein the second antigen-binding domain that binds to TfR comprises a heavy chain CDR1 (HCDR1) domain having an amino acid sequence selected from SEQ ID NOs: 272, 283, 294, 304, 315, 327, 339, 351, 362, 373, 383, 394, 405, 415, 426, 437, 448, 458, 469, 481, 492, 502, 512, 522, 532, 543, 553, 563, 573, 583, 594, 605, or 1121, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; a heavy chain CDR2 (HCDR2) domain having an amino acid sequence selected from SEQ ID NOs: 273, 284, 295, 305, 316, 328, 340, 352, 363, 374, 384, 395, 406, 416, 427, 4
  • the anti-TfR antibodies, or antigen-binding fragments ⁇ ⁇ Attorney Docket No.250298.000954 thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014, 1015, 1375, 1016, 1387, 1017, 1019, 1381, 1020, 1021, 1384, 1022, 1023, 1024, 1025, or 1027, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996
  • the nucleotide sequence that encodes the anti-TfR antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014, 1015, 1375, 1016, 1387, 1017, 1019, 1381, 1020, 1021, 1384, 1022, 1023, 1024, 1025, or 1027, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000,
  • the anti-TfR antibodies, or antigen-binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NOs: 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014, 1015, 1375, 1016, 1387, 1017, 1019, 1381, 1020, 1021, 1384, 1022, 1023, 1024, 1025, or 1027.
  • the anti-TfR antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 975, 977, 979, 981, 983, 985, 987, 989, 991, 993, 995, 997, 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 20, 1018, 1026, or 1028, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 975, 977, 979, 981, 983, 985, 987, 989, 991, 993, 995, 997, 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 20, 1018, 1026, or 1028.
  • the nucleotide sequence that encodes the anti-TfR antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 975, 977, 979, 981, 983, 985, 987, 989, 991, 993, 995, 997, 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 20, 1018, 1026, or 1028, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 975, 977, 979, 981, 983, 985, 987, 989, 991, 993, 995, 997, 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 20, 1018, 1026, or 1028.
  • the anti-TfR antibodies ⁇ ⁇ Attorney Docket No.250298.000954 or antigen-binding fragments thereof, comprise the amino acid sequence set forth in SEQ ID NOs: 975, 977, 979, 981, 983, 985, 987, 989, 991, 993, 995, 997, 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 20, 1018, 1026, or 1028.
  • Certain non-limiting, exemplary anti-AAV/anti-TfR multispecific antibodies provided herein include a second antigen-binding domain that binds to TfR comprising a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3), respectively, having the amino acid sequences selected from SEQ ID NOs: 272- 273- 274- 277- 278- 279 (e.g., REGN31874B), SEQ ID NOs: 283- 284- 285- 288- 289- 290 (e.g., REGN31863B), SEQ ID NOs: 294- 295- 296- 299- 300- 301 (e.g., REGN69348), SEQ ID NOs: 304- 305- 306- 310- 311- 312 (e.g., REGN69340), SEQ ID NOs: 315- 316- 317- 321- 322- 3
  • an anti-AAV/anti-TfR multispecific antibody herein can comprise an anti-TfR antibody, or antigen-binding fragment thereof, wherein the anti-TfR antibody or antigen-binding fragment thereof can include: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and ⁇ ⁇ Attorney Docket No.250298.000954 HCDR3 contained within a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a light chain variable region (LCVR) that comprises: a heavy chain variable region (HCVR)
  • an anti-AAV/anti-TfR multispecific antibody herein can comprise an anti-TfR antibody or antigen-binding fragment thereof, wherein the anti-TfR antibody or antigen-binding fragment thereof, can include: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the ⁇ ⁇ Attorney Docket No.250298.000954 amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and
  • an anti-AAV/anti-TfR multispecific antibody herein can comprise an anti-TfR antibody, or antigen-binding fragment thereof, wherein the anti-TfR antibody or antigen-binding fragment thereof can include: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 30
  • an anti-AAV/anti-TfR multispecific antibody herein can comprise an anti-TfR antigen-binding fragment comprising an anti-TfR Fab, wherein the anti-TfR Fab can include: a) a HC region that comprises the amino acid sequence of SEQ ID NO: 974, or a variant thereof; and/or a LC region that comprises the amino acid sequence of SEQ ID NO: 975, or a variant thereof; b) a HC region that comprises the amino acid sequence of SEQ ID NO: 976, or a variant thereof; and/or a LC region that comprises the amino acid sequence of SEQ ID NO: 977, or a variant thereof; c) a HC region that comprises the amino acid sequence of SEQ ID NO: 978, or a variant thereof; and/or a LC region that comprises the amino acid sequence of SEQ ID NO: 979, or a variant thereof; d) a HC region that comprises the amino acid sequence of
  • any of the HCVR and/or LCVR of the anti-TfR antibodies, or variants thereof, disclosed herein can be contained within an antigen-binding fragment such as but not limited to (i) Fab fragment; (ii) F(ab')2 fragment; (iii) Fd fragment; (iv) Fv fragment; (v) single-chain Fv (scFv) molecules; and (vi) dAb fragments.
  • an antigen-binding fragment such as but not limited to (i) Fab fragment; (ii) F(ab')2 fragment; (iii) Fd fragment; (iv) Fv fragment; (v) single-chain Fv (scFv) molecules; and (vi) dAb fragments.
  • HCVR and LCVR of an anti-TfR antibody are contained within an antigen binding fragment (e.g., an scFv or a Fab)
  • the HCVR and the LCVR can be in either orientation (HCVR-LCVR or LCVR- HCVR).
  • the HCVR and LCVR are optionally linked by a linker, e.g., that comprises an amino acid sequence, e.g., about 10 amino acids in length, for example: (Gly4Ser)n (SEQ ID NO: 241) wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the scFv comprises (i) a heavy chain variable region that comprises the HCDR1, HCDR2, and HCDR3 of a HCVR comprising the amino acid sequence of SEQ ID NO: 270, 271, 281, 282, 292, 293, 303, 314, 325, 326, 337, 338, 349, 350, 361, 371, 372, 382, 392, 393, 404, 414, 424, 425, 436, 446, 447, 457, 467, 468, 479, 480, 490, 491, 501, 511, 521, 531, 541, 542, 552, 562, 572, 582, 592, 593, 604, or 1119; and/or (ii) a light chain variable region that comprises the LCDR1, LCDR2, and LCDR3 of a LCVR comprising the amino acid sequence of SEQ ID NO: 276, 287, 298, 308, 309, 319, 320, 331, 332, 343, 3
  • an anti-TfR scFv of the present disclosure in VL-(Gly4Ser)3 (SEQ ID NO: 1115)-VH format, comprises an amino acid sequence as of Table 9.
  • the present disclosure includes scFvs that are in the format VH- (Gly4Ser)3(SEQ ID NO: 1115)-VL. Table 9.
  • Anti-hTfR scFv Molecules ⁇ ⁇ Attorney Docket No.250298.000954 ⁇ ⁇ Attorney Docket No.250298.000954 ⁇ ⁇ Attorney Docket No.250298.000954 ⁇ ⁇ Attorney Docket No.250298.000954 [00568] Fab fragments that bind specifically to TfR form part of the present disclosure.
  • Fab fragments typically contain one complete light chain, VL and a constant light (CL) domain, e.g., kappa (e.g., RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 1147)); and the VH and IgG1 CH1 portion (e.g., ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH (SEQ ID NO: 1179)) or ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK
  • Fab fragment antibodies can be generated by papain digestion of whole IgG antibodies to remove the entire Fc fragment, including the hinge region.
  • the present disclosure includes Fab proteins comprising: (1) a heavy chain variable region (HCVR) that comprises the amino acid sequence of SEQ ID NO: 270 or 271, or a heavy chain variable region that includes HCDR1, HCDR2, and HCDR3 of such a HCVR- linked to the CH1 domain; and a light chain variable region (LCVR) that comprises the amino acid sequence of SEQ ID NO: 276, or LCDR1, LCDR2, and LCDR3 of such a LCVR-linked to the CL domain; (2) a heavy chain variable region (HCVR) that comprises the amino acid sequence of SEQ ID NO: 281 or 282, or a heavy chain variable region that includes HCDR1, HCDR2, and HCDR3 of such a HCVR- linked to the CH1 domain; and a light chain variable region (LCVR) that comprises the amino acid sequence of SEQ ID NO: 287, or LCDR1, LCDR
  • an anti-AAV/anti- CACNG1 multispecific antibody may comprise a second antigen-binding domain that binds to CACNG1, wherein the second antigen-binding domain comprises any of the heavy chain variable region (HCVR) amino acid sequences as set forth in Table 10.
  • the second antigen- binding domain that binds to CACNG1 may also comprise any of the light chain variable region (LCVR) amino acid sequences as set forth in Table 10.
  • the second antigen-binding domain that binds to CACNG1 may comprise any of the HCVR/LCVR amino acid sequence pairs as set forth in Table 10.
  • the present disclosure also provides anti-AAV/anti-CACNG1 multispecific antibodies, wherein the second antigen- binding domain that binds to CACNG1 comprises any of the heavy chain CDR1-CDR2- CDR3 amino acid sequences as set forth in Table 10, and/or any of the light chain CDR1- CDR2-CDR3 amino acid sequences as set forth in Table 10.
  • nucleic acid molecules encoding any of the CDR, HCVR, LCVR, HC, or LC sequences of the anti-AAV/anti-CACNG1 multispecific antigen-binding molecules disclosed herein, or variants thereof, including nucleic acid ⁇ ⁇ Attorney Docket No.250298.000954 molecules comprising any of the polynucleotide sequences as set forth in Table 2 herein, as well as nucleic acid molecules comprising any of the polynucleotide sequences as set forth in Table 11 herein, in any combination thereof.
  • the nucleic acid molecules described herein may comprise one or more polynucleotide sequence(s) in Table 2 that encode a pair of HCVR/LCVR amino acid sequences, and one or more polynucleotide sequence(s) selected from Table 11 that encode a pair of HCVR/LCVR amino acid sequences.
  • the nucleic acid molecules described herein may comprise one or more polynucleotide sequence(s) selected from Table 2 that encode a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3), and one or more polynucleotide sequence(s) selected from Table 11 that encode a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3).
  • Table 10 Amino Acid Sequence Identifiers for Anti-CACNG1 Antibodies ⁇ ⁇ Attorney Docket No.250298.000954 Table 11.
  • anti-AAV/anti-CACNG1 multispecific antibodies wherein the second antigen-binding domain that binds to CACNG1 comprises a light chain variable region (LCVR) having an amino acid sequence selected from SEQ ID NOs: 623, 643, 663, 683, 703, 723, 743, 763, 783, 803, 823, 843, 863, 883, 903, 923, 943, 963, 1189, or 1213 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • LCVR light chain variable region
  • anti-AAV/anti-CACNG1 multispecific antibodies wherein the second antigen-binding domain that binds to CACNG1 comprises a HCVR and LCVR (HCVR/LCVR) amino acid sequence pair selected from SEQ ID NOs: 615/623 (e.g., H2aM31929N/REGN10728), SEQ ID NOs: 635/643 (e.g., H2aM31944N), SEQ ID NOs: 655/663 (e.g., H4H31265P2/REGN5972), SEQ ID NOs: 675/683 (e.g., H2aM31941N), SEQ ID NOs: 695/703 (e.g., REGN7660), SEQ ID NOs: 715/723 (e.g., REGN9909), SEQ ID NOs: 735/743 (e.g., REGN10713), SEQ ID NOs: 755/763 (e
  • anti-AAV/anti-CACNG1 multispecific ⁇ ⁇ Attorney Docket No.250298.000954 antibodies wherein the second antigen-binding domain that binds to CACNG1 comprises a heavy chain CDR1 (HCDR1) domain having an amino acid sequence selected from SEQ ID NOs: 617, 637, 657, 677, 697, 717, 737, 757, 777, 797, 817, 837, 857, 877, 897, 917, 937, 957, 1183, or 1207 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; a heavy chain CDR2 (HCDR2) domain having an amino acid sequence selected from SEQ ID NOs: 619, 639, 659, 679, 699, 719, 739, 759, 779, 799, 819, 839, 859, 879, 899, 919, 939
  • the anti-CACNG1 antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 631, 651, 671, 691, 711, 731, 751, 771, 791, 811, 1342, 1369, 831, 851, 871, 891, 911, 931, 951, 971, 1197, or 1221, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 631, 651, 671, 691, 711, 731, 751, 771, 791, 811, 1342, 1369, 831, 851, 871, 891, 911, 931, 951, 971, 1197, or 1221.
  • the nucleotide sequence that encodes the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 631, 651, 671, 691, 711, 731, 751, 771, 791, 811, 1342, 1369, 831, 851, 871, 891, 911, 931, 951, 971, 1197, or 1221, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 631, 651, 671, 691, 711, 731, 751, ⁇ ⁇ Attorney Docket No.250298.000954 771, 791, 811, 1342, 1369, 831, 851, 871, 891, 911, 931, 951, 971, 1197, or 1221.
  • the nucleotide sequence that encodes the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 630, 650, 670, 690, 710, 730, 750, 770, 790, 810, 830, 850, 870, 890, 910, 930, 950, 970, 1196, or 1220, or a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the nucleotide sequence of SEQ ID NOs: 630, 650, 670, 690, 710, 730, 750, 770, 790, 810, 830, 850, 870, 890, 910, 930, 950, 970, 1196, or 1220.
  • the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NOs: 631, 651, 671, 691, 711, 731, 751, 771, 791, 811, 1342, 1369, 831, 851, 871, 891, 911, 931, 951, 971, 1197, or 1221.
  • the nucleotide sequence that encodes the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 630, 650, 670, 690, 710, 730, 750, 770, 790, 810, 830, 850, 870, 890, 910, 930, 950, 970, 1196, or 1220.
  • the anti-CACNG1 antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 633, 653, 673, 693, 713, 733, 753, 773, 793, 813, 20, 833, 853, 873, 893, 913, 933, 953, 973, 1201, or 1223, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 633, 653, 673, 693, 713, 733, 753, 773, 793, 813, 20, 833, 853, 873, 893, 913, 933, 953, 973, 1201, or 1223.
  • the nucleotide sequence that encodes the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 633, 653, 673, 693, 713, 733, 753, 773, 793, 813, 20, 833, 853, 873, 893, 913, 933, 953, 973, 1201, or 1223, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs: 633, 653, 673, 693, 713, 733, 753, 773, 793, 813, 20, 833, 853, 873, 893, 913, 933, 953, 973, 1201, or 1223.
  • the nucleotide sequence that encodes the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 632, 652, 672, 692, 712, 732, 752, 772, 792, 812, 1346, 1368, 19, 832, 852, 872, 892, 912, 932, 952, 972, 1200, or 1224, or a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the nucleotide sequence of SEQ ID NOs: 632, 652, 672, 692, 712, 732, 752, 772, 792, 812, 1346, 1368, 19, 832, 852, 872, 892, 912, 932, 952, 972, 1200, or 1224.
  • the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NOs: 633, 653, 673, 693, ⁇ ⁇ Attorney Docket No.250298.000954 713, 733, 753, 773, 793, 813, 20, 833, 853, 873, 893, 913, 933, 953, 973, 1201, or 1223.
  • the nucleotide sequence that encodes the anti-CACNG1 antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 632, 652, 672, 692, 712, 732, 752, 772, 792, 812, 1346, 1368, 19, 832, 852, 872, 892, 912, 932, 952, 972, 1200, or 1224.
  • Certain non-limiting, exemplary anti-AAV/anti-CACNG1 multispecific antibodies provided herein include a second antigen-binding domain that binds to CACNG1 comprising a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3), respectively, having the amino acid sequences selected from SEQ ID NOs: 617- 619- 621- 625- 627- 629 (e.g., H2aM31929N/REGN10728), SEQ ID NOs: 637- 639- 641- 645- 647- 649 (e.g., H2aM31944N), SEQ ID NOs: 657- 659- 661- 665- 667- 669 (e.g., H4H31265P2/REGN5972), SEQ ID NOs: 677- 679- 681- 685- 687- 689 (e.g., H2aM319
  • an anti-AAV/anti-CACNG1 multispecific antibody herein can comprise an anti-CACNG1 antibody, or antigen-binding fragment thereof, wherein the anti-CACNG1 antibody or antigen-binding fragment thereof can include: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a HCVR comprising the amino acid sequence of SEQ
  • an anti-AAV/anti-CACNG1 multispecific antibody herein can comprise an anti-CACNG1 antibody or antigen-binding fragment thereof, wherein the anti-CACNG1 antibody or antigen-binding fragment thereof, can include: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of S
  • an anti-AAV/anti-CACNG1 multispecific antibody herein can comprise an anti-CACNG1 antibody, or antigen-binding fragment thereof, wherein the anti-CACNG1 antibody or antigen-binding fragment thereof can include: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR ⁇ ⁇ Attorney Docket No.250298.000954 comprising the amino acid sequence of SEQ ID NO: 623; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 643; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR
  • an anti-AAV/anti-CACNG1 multispecific antibody herein can comprise an anti-CACNG1 antigen-binding fragment comprising an anti- CACNG1 Fab, wherein the anti-CACNG1 Fab can include: a) a HC region that comprises the amino acid sequence of SEQ ID NO: 631, or a variant thereof; and/or a LC region that comprises the amino acid sequence of SEQ ID NO: 633, or a variant thereof; b) a HC region that comprises the amino acid sequence of SEQ ID NO: 651, or a variant thereof; and/or a LC region that comprises the amino acid sequence of SEQ ID NO: 653, or a variant thereof; c) a HC region that comprises the amino acid sequence of SEQ ID NO: 671, or a variant thereof; and/or a LC region that comprises the amino acid sequence of SEQ ID NO: 673, or a variant thereof; d) a HC region that comprises the amino acid
  • any of the HCVR and/or LCVR of the anti-CACNG1 antibodies, or variants thereof, disclosed herein can be contained within an antigen-binding fragment such as but not limited to (i) Fab fragment; (ii) F(ab')2 fragment; (iii) Fd fragment; (iv) Fv fragment; (v) single-chain Fv (scFv) molecules; and (vi) dAb fragments.
  • an antigen-binding fragment such as but not limited to (i) Fab fragment; (ii) F(ab')2 fragment; (iii) Fd fragment; (iv) Fv fragment; (v) single-chain Fv (scFv) molecules; and (vi) dAb fragments.
  • HCVR and LCVR of an anti- CACNG1 antibody, or variant thereof are contained within an antigen binding fragment (e.g., an scFv or a Fab)
  • the HCVR and the LCVR can be in either orientation (HCVR-LCVR or LCVR-HCVR).
  • the HCVR and LCVR are optionally linked by a linker, e.g., that comprises an amino acid sequence, e.g., about 10 amino acids in length, for example: (Gly4Ser)n (SEQ ID NO: 241) wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the scFv comprises (i) a heavy chain variable region that comprises the HCDR1, HCDR2, and HCDR3 of a HCVR comprising the amino acid ⁇ ⁇ Attorney Docket No.250298.000954 sequence of SEQ ID NO: 615, 635, 655, 675, 695, 715, 735, 755, 775, 795, 815, 835, 855, 875, 895, 915, 935, 955, 1181, or 1205; and/or (ii) a light chain variable region that comprises the LCDR1, LCDR2, and LCDR3 of a LCVR comprising the amino acid sequence of SEQ ID NO: 623, 643, 663, 683, 703, 723, 743, 763, 783, 803, 823, 843, 863, 883, 903, 923, 943, 963, 1189, or 1213; or the scFv comprises: (1) a HCVR comprising the
  • an scFv fragment that binds specifically to CACNG1 comprises the sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGCGTKVEIKGGGGSGGGGSGGG GSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKCLEWVAVI WHDGSDKYYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGL GTLVTVSS (SEQ ID NO: 1355), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • an scFv fragment that binds specifically to CACNG1 comprises the sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKCLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS GGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQ KPGKAPNLLIYKASNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFG CGTKVEIK (SEQ ID NO: 1350), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab fragments that bind specifically to CACNG1 form part of the present disclosure.
  • Fab fragments typically contain one complete light chain, VL and a constant light (CL) domain, e.g., kappa (e.g., RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 1147)); and the VH and IgG1 CH1 portion (e.g., ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG ⁇ ⁇ Attorney Docket No.250298.000954 LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH (SEQ ID NO: 1179) or ASTKGPSVFPLAPSSKSTSGGTAAL
  • Fab fragment antibodies can be generated by papain digestion of whole IgG antibodies to remove the entire Fc fragment, including the hinge region.
  • the present disclosure includes Fab proteins comprising: (1) a heavy chain variable region (HCVR) that comprises the amino acid sequence of SEQ ID NO: 615, or a heavy chain variable region that includes HCDR1, HCDR2, and HCDR3 of such a HCVR- linked to the CH1 domain; and a light chain variable region (LCVR) that comprises the amino acid sequence of SEQ ID NO: 623, or LCDR1, LCDR2, and LCDR3 of such a LCVR-linked to the CL domain; (2) a heavy chain variable region (HCVR) that comprises the amino acid sequence of SEQ ID NO: 635, or a heavy chain variable region that includes HCDR1, HCDR2, and HCDR3 of such a HCVR- linked to the CH1 domain; and a light chain variable region (LCVR) that comprises the amino acid sequence of SEQ ID NO: 643, or LCDR1, LCDR2, and LCDR
  • the isolated bispecific antigen-binding molecule competes for binding to a molecule on a cell surface, or binds to the same epitope on a molecule on a cell surface as a reference antibody, wherein the reference antibody comprises an antibody or antigen-binding fragment thereof, or a bispecific antibody disclosed herein.
  • the bispecific antibody comprises a human IgG heavy chain constant region. In some cases, the human IgG heavy chain constant region is isotype IgG1. In some cases, the human IgG heavy chain constant region is isotype IgG4.
  • the bispecific antibody comprises a chimeric hinge that reduces Fc ⁇ receptor binding relative to a wild-type hinge of the same isotype.
  • the first antigen-binding domain and the second antigen-binding domain may be directly or indirectly connected to one another to form a bispecific antigen-binding molecule of the present disclosure.
  • the first antigen-binding domain and the second antigen-binding domain may each be connected to a separate multimerizing domain. The association of one multimerizing domain with another multimerizing domain facilitates the association between the two antigen-binding domains, thereby forming a bispecific antigen- binding molecule.
  • a “multimerizing domain” is any macromolecule, protein, polypeptide, peptide, or amino acid that has the ability to associate with a second multimerizing domain of the same or similar structure or constitution.
  • a multimerizing domain may be a polypeptide comprising an immunoglobulin CH3 domain.
  • a non-limiting example of a multimerizing component is an Fc portion of an immunoglobulin (comprising a C H 2-C H 3 domain), e.g., an Fc domain of an IgG selected from the isotypes IgG1, IgG2, IgG3, and IgG4, as well as any allotype within each isotype group.
  • Bispecific antigen-binding molecules of the present disclosure will typically comprise two multimerizing domains, e.g., two Fc domains that are each individually part of a separate antibody heavy chain.
  • the first and second multimerizing domains may be of the same IgG isotype such as, e.g., IgG1/IgG1, IgG2/IgG2, IgG4/IgG4.
  • the first and second multimerizing domains may be of different IgG isotypes such as, e.g., IgG1/IgG2, IgG1/IgG4, IgG2/IgG4, etc.
  • the multimerizing domain is an Fc fragment or an amino acid sequence of from 1 to about 200 amino acids in length containing at least one cysteine residue. In other embodiments, the multimerizing domain is a cysteine residue, or a short cysteine-containing peptide.
  • Other multimerizing domains include peptides or polypeptides comprising or consisting of a leucine zipper, a helix-loop motif, or a coiled-coil motif.
  • the multispecific (e.g., bispecific) antigen-binding molecule (e.g., antibody or antigen-binding fragment thereof) that binds to a capsid of an AAV particle and a molecule on a cell surface includes: a) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 4, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 6, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 8, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: b) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 24, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 26, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 28, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant thereof; and a second antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: c) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 34, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 36, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 38, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant thereof; and
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: d) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 44, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 46, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 48, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant thereof; and
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: e) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 54, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 56, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 58, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant thereof;
  • a first antigen binding domain that binds to a capsid of an AAV that comprises: a
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: f) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 64, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 56, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 67, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant thereof; and
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: h) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 83, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 85, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 87, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant thereof
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: i) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 93, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 95, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 97, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant thereof
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: j) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 103, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 105, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 107, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 12, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 14, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 16, or a variant
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: k) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 113, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 115, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 117, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO:
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: l) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 133, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 135, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 137, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 125
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: m) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 143, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 145, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 147, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO:
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: n) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 153, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 155, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 157, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO:
  • the multispecific (e.g., bispecific) antigen-binding molecule (e.g., antibody or antigen-binding fragment thereof) that binds to a capsid of an AAV particle and a molecule on a cell surface includes: [00610] o) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 163, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 165, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 167, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: p) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 173, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 145, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 176, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 125
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: q) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 182, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 184, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 186, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 125
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: r) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 192, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 194, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 196, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 125
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: s) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 202, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 204, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 206, or a variant thereof; a LCDR1 that comprises the amino acid sequence of SEQ ID NO: 121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO:
  • the multispecific (e.g., bispecific) antigen-binding molecule that binds to a capsid of an AAV particle and a molecule on a cell surface includes: t) (1) a first antigen binding domain that binds to a capsid of an AAV that comprises: a HCDR1 that comprises the amino acid sequence of SEQ ID NO: 212, or a variant thereof; a HCDR2 that comprises the amino acid sequence of SEQ ID NO: 214, or a variant thereof; a HCDR3 that comprises the amino acid sequence of SEQ ID NO: 216121, or a variant thereof; a LCDR2 that comprises the amino acid sequence of SEQ ID NO: 123, or a variant thereof; and a LCDR3 that comprises the amino acid sequence SEQ ID NO: 125, or a variant thereof; and a second antigen binding domain that binds to a molecule
  • Any multispecific antibody (e.g., bispecific antibody) format or technology may be used to make the multispecific antigen-binding molecules of the present disclosure.
  • an antibody or fragment thereof having a first antigen-binding specificity can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association, or otherwise) to one or more other molecular entities, such as another antibody or antibody fragment having a second antigen-binding specificity to produce a bispecific antigen-binding molecule.
  • Specific exemplary multispecific formats that can be used in the context of the present disclosure include, without limitation, e.g., scFv-based, VHH-based, or diabody bispecific formats, IgG-scFv fusions, dual variable domain (DVD)-Ig, Quadroma, knobs-into- holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, ⁇ ⁇ Attorney Docket No.250298.000954 CrossFab, (SEED)body, leucine zipper, Duobody, IgG1/IgG2, dual acting Fab (DAF)-IgG, and Mab 2 bispecific formats (see, e.g., Klein et al.2012, mAbs 4:6, 1-11, and references cited therein, for a review of the foregoing formats).
  • the multimerizing domains may comprise one or more amino acid changes (e.g., insertions, deletions, or substitutions) as compared to the wild-type, naturally occurring version of the Fc domain.
  • the disclosure includes bispecific antigen- binding molecules comprising one or more modifications in the Fc domain that results in a modified Fc domain having a modified binding interaction (e.g., enhanced or diminished) between Fc and FcRn.
  • the bispecific antigen-binding molecule comprises a modification in a CH2 or a CH3 region, wherein the modification increases the affinity of the Fc domain to FcRn in an acidic environment (e.g., in an endosome where pH ranges from about 5.5 to about 6.0).
  • Non-limiting examples of such Fc modifications include, e.g., a modification at position 250 (e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., L/Y/F/W or T), 254 (e.g., S or T), and 256 (e.g., S/R/Q/E/D or T); or a modification at position 428 and/or 433 (e.g., L/R/S/P/Q or K) and/or 434 (e.g., H/F or Y); or a modification at position 250 and/or 428; or a modification at position 307 or 308 (e.g., 308F, V308F), and 434.
  • a modification at position 250 e.g., E or Q
  • 250 and 428 e.g., L or F
  • 252 e.g., L/Y/F/W or T
  • 254 e.g., S or T
  • the modification comprises a 428L (e.g., M428L) and 434S (e.g., N434S) modification; a 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modification; a 433K (e.g., H433K) and a 434 (e.g., 434Y) modification; a 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modification; a 250Q and 428L modification (e.g., T250Q and M428L); and a 307 and/or 308 modification (e.g., 308F or 308P).
  • a 428L e.g., M428L
  • 434S e.g., N434S
  • 428L, 259I e.g., V259I
  • 308F e.g., V308F
  • the present disclosure also includes multispecific antigen-binding molecules comprising a first CH3 domain and a second Ig CH3 domain, wherein the first and second Ig CH3 domains differ from one another by at least one amino acid, and wherein at least one amino acid difference reduces binding of the bispecific antibody to Protein A as compared to a bi-specific antibody lacking the amino acid difference.
  • the first Ig CH3 domain binds Protein A and the second Ig C H 3 domain contains a mutation that reduces or abolishes Protein A binding such as an H95R modification (by IMGT exon numbering; H435R by EU numbering).
  • the second CH3 may further comprise a Y96F modification (by IMGT; Y436F by EU).
  • the Fc domain may be chimeric, combining Fc sequences derived from more than one immunoglobulin isotype.
  • a chimeric Fc domain can comprise part or all of a CH2 sequence derived from a human IgG1, human IgG2, or human IgG4 C H 2 region, and part or all of a C H 3 sequence derived from a human IgG1, human IgG2, or human IgG4.
  • a chimeric Fc domain can also contain a chimeric hinge region.
  • a chimeric hinge may comprise an “upper hinge” sequence, derived from a human IgG1, a human IgG2, or a human IgG4 hinge region, combined with a “lower hinge” sequence, derived from a human IgG1, a human IgG2, or a human IgG4 hinge region.
  • a particular example of a chimeric Fc domain that can be included in any of the antigen-binding molecules set forth herein comprises, from N- to C-terminus: [IgG4 C H 1] - [IgG4 upper hinge] - [IgG2 lower hinge] - [IgG4 CH2] - [IgG4 CH3].
  • chimeric Fc domains that can be included in any of the antigen-binding molecules of the present disclosure are described in US Publication 2014/0243504, published August 28, 2014, which is herein incorporated in its entirety. Chimeric Fc domains having these general structural arrangements, and variants thereof, can have altered Fc receptor binding, which in turn affects Fc effector function.
  • Example alternative antibody formats are described in Figures 3, 9, 18A-18B, 29A-29I, 30A-30G, 36A-36B, 46A-46I, and 47A-47I.
  • Alternative Format Antibodies [00621] The present disclosure provides any of the multispecific antibodies, or antigen binding fragments thereof, described herein in any of various alternative formats described herein.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) a first heavy chain region of a first Fab (“Fab1”), the first heavy chain region operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and ⁇ ⁇ Attorney Docket No.250298.000954 d)
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the scFv is linked to the first heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, the Fc domain operably linked to (iii) an scFv comprising a first antigen-binding domain (“ABD1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d) a fourth polypeptide chain comprising a second light chain that pairs with
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the scFv is linked to the Fc domain via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”), the second heavy chain region of Fab2 operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth antigen-binding domain
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the first heavy chain region is linked to the second heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, the Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second ⁇ ⁇ Attorney
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an scFv comprising a first antigen- binding domain (“ABD1”), the scFv operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d) a fourth polypeptide chain comprising a second light chain that pairs
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ⁇ ⁇ Attorney Docket No.250298.000954 ABD3 binds to a molecule on a cell surface.
  • the scFv is linked to the first heavy chain region via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, the Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • Fab2 is linked to the Fc domain via a linker.
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO:
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ⁇ ⁇ Attorney Docket No.250298.000954 ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 or 1159, and/or a LCVR comprising the amino acid sequence
  • the molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated ⁇ ⁇ Attorney Docket No.250298.000954 channel auxiliary subunit gamma 1 (CACNG1).
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated ⁇ ⁇ Attorney Docket No.250298.000954 channel auxiliary subunit gamma 1
  • CACNG1 channel auxiliary subunit gamma 1
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 222, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 226; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 232, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 234, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 236; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises a) a HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the molecule on the cell surface is TfR.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that ⁇ ⁇ Attorney Docket No.250298.000954 comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of S
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 288, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 27
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 314, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 31
  • the molecule on the cell surface is CACNG1.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR that comprises the HCDR1, HCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 6
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 645, a
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/or a LCVR comprising the amino acid sequence of SEQ ID NO:
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); and c) a third polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form Fab2, wherein Fab2 comprises a second antigen-binding domain (“ABD2”); wherein ABD1 or ABD2 binds to a capsid of an adeno-associated virus (AAV) particle, and the other of ABD1 or ABD2 binds to
  • ABD1 binds to a capsid of an adeno-associated virus (AAV) particle and ABD2 binds to a molecule on the cell surface.
  • ABD2 binds to a capsid of an adeno-associated virus (AAV) particle and ABD1 binds to a molecule on the cell surface.
  • the first heavy chain region is linked to the second heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: ⁇ ⁇ Attorney Docket No.250298.000954 a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) an Fc domain; b) a second polypeptide chain, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain; and c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); wherein ABD1 or ABD2 binds to a capsid of an adeno-associated virus (AAV) particle, and
  • ABD1 binds to a capsid of an adeno-associated virus (AAV) particle and ABD2 binds to a molecule on the cell surface.
  • ABD2 binds to a capsid of an adeno-associated virus (AAV) particle and ABD1 binds to a molecule on the cell surface.
  • the first scFv and/or the first heavy chain region is linked to the Fc domain via a linker.
  • the ABD1 or ABD2 which binds to a capsid of an AAV particle comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and
  • the ABD1 or ABD2 which binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO:
  • the ABD1 or ABD2 which binds to a capsid of an AAV particle comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 or 1159, and/or a
  • the molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary subunit gamma 1 (CACNG1). [00680] In some embodiments, the molecule on the cell surface is ASGR1.
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • the other of ABD1 or ABD2 which binds to ASGR1 comprises: a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the other of ABD1 or ABD2 which binds to ASGR1 comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 222, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 226; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 232, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 234, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 236; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO:
  • the other of ABD1 or ABD2 which binds to TfR comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or
  • the other of ABD1 or ABD2 which binds to TfR comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 288, a LCDR2 comprising the amino acid sequence of SEQ ID NO:
  • the other of ABD1 or ABD2 which binds to TfR comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR ⁇ ⁇ Attorney Docket No.250298.000954 comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 314, and
  • the molecule on the cell surface is CACNG1.
  • the other of ABD1 or ABD2 which binds to CACNG1 comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 643; ⁇ ⁇ Attorney Docket No.250298.000954 3) a HCVR that comprises the HCDR1,
  • the other of ABD1 or ABD2 which binds to CACNG1 comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID
  • the other of ABD1 or ABD2 which binds to CACNG1 comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR ⁇ ⁇ Attorney Docket No.250298.000954 comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/or LCVR comprising
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) a first heavy chain region of a first Fab (“Fab1”), said first heavy chain region operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d) a fourth polypeptide chain comprising a second light chain that pairs
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the scFv is linked to the first heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) an scFv comprising a first antigen-binding domain (“ABD1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and ⁇ ⁇ Attorney Docket No.250298.000954 d)
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the scFv is linked to the Fc domain via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”), said second heavy chain region of Fab2 operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the first heavy chain region is linked to the second heavy chain region via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an scFv comprising a first antigen-binding domain (“ABD1”), said scFv operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) ⁇ ⁇ Attorney Docket No.250298.000954 a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the scFv is linked to the first heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d) a fourth polypeptide chain comprising a second light chain that pairs with
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV ⁇ ⁇ Attorney Docket No.250298.000954 particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the first heavy chain region is operably linked to the second heavy chain region and/or the second heavy chain region is operably linked to the third heavy chain region via a linker.
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, ⁇ ⁇ Attorney Docket No.250298.000954 and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 4,
  • said molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary subunit gamma 1 (CACNG1). [00731] In some embodiments, said molecule on the cell surface is ASGR1.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises a) a HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR ⁇ ⁇ Attorney Docket No.250298.000954 comprising the amino acid sequence of SEQ ID NO: 10.
  • the molecule on the cell surface is TfR.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 292 or
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 ⁇ ⁇ Attorney Docket No.250298.000954 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO:
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 314, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 31
  • the molecule on the cell surface is CACNG1.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 6
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 645,
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) a first heavy chain region of a first Fab (“Fab1”), the first heavy chain region of Fab1 operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second scFv comprising a second antigen-binding domain (“ABD2”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”), the second heavy chain region of Fab2 operably linked to (iii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD3 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD2 binds to a molecule on a cell surface.
  • the first scFv is linked to the first heavy chain region and/or the second scFv is linked to the second heavy chain region via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy ⁇ ⁇ Attorney Docket No.250298.000954 chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, the Fc domain operably linked to (iii) a first scFv comprising a first antigen-binding domain (“ABD1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain, the Fc domain oper
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD3 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD2 binds to a molecule on a cell surface.
  • the first scFv and/or second scFv is linked to the Fc domain via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”) operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operable liked to (ii) a fourth heavy chain region of a fourth Fab (“Fab4”) operably linked to (iii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 and ABD4 binds to a molecule on a cell surface.
  • ABD2 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD3 binds to a molecule on a cell surface.
  • the first heavy chain region is linked to the second heavy chain region and/or the third heavy chain region is linked to fourth heavy chain region via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a fourth heavy chain region of a fourth Fab (“Fab4”); c)
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an ⁇ ⁇ Attorney Docket No.250298.000954 AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD2 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD3 binds to a molecule on a cell surface.
  • the first heavy chain region, the second heavy chain region, the third heavy chain region, and/or the fourth heavy chain region is linked to the Fc domain via a linker.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of the second Fab and the fourth Fab are in a CrossmAb arrangement.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of the first Fab and the third Fab are in a CrossmAb arrangement.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of the first Fab and the second Fab are in a CrossmAb arrangement.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of the third Fab and the fourth Fab are in a CrossmAb arrangement.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of the first Fab and the fourth Fab are in a CrossmAb arrangement.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of the second Fab and the third Fab are in a CrossmAb arrangement.
  • one or more light chains comprise the same light chain variable region.
  • the first light chain, the second light chain, the third light chain, and/or the fourth light chain comprise the same light chain variable region.
  • the first light chain, the second light chain, the third light chain, and/or the fourth light chain are universal light chains.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to a first heavy chain region of a first Fab (“Fab1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second scFv comprising a second antigen-binding domain (“ABD2”) operably linked to (ii) an Fc domain, said Fc domain operably linked to a second heavy chain region of a second Fab (“Fab2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a third antigen-
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD3 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD2 binds to a molecule on a cell surface.
  • the at least one of ABD1, ABD2, ABD3, and ABD4 which binds to a capsid of an AAV particle comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained
  • the at least one of ABD1, ABD2, ABD3, and ABD4 which binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID
  • the molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary subunit gamma 1 (CACNG1).
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to ASGR1 comprises a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the ⁇ ⁇ Attorney Docket No.250298.000954 LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to ASGR1 comprises a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 222, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 226; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 232, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 234, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 236; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCD
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to ASGR1 comprises a) a HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the molecule on the cell surface is TfR.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to TfR comprises 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO:
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to TfR comprises 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 288, a LCDR2 comprising the amino acid sequence of SEQ ID
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds ⁇ ⁇ Attorney Docket No.250298.000954 to TfR comprises 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of
  • the molecule on the cell surface is CACNG1.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to CACNG1 comprises 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR ⁇ ⁇ Attorney Docket No.250298.000954 comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising 1)
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to CACNG1 comprises 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 645
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to CACNG1 comprises 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/or a LCVR comprising the amino acid sequence of S
  • the linker is: (a) at least 5 amino acids, at least 6 amino acids, or at least 7 amino acids in length; and optionally (b) up to 30 amino acids, up to 40 amino acids, up to 50 amino acids, or up to 60 amino acids in length.
  • the linker is: (a) 5 amino acids to 50 amino acids in length; (b) 5 amino acids to 45 amino acids in length; (c) 5 amino acids to 40 amino acids in length; (d) 5 amino acids to 35 amino acids in length; (e) 5 amino acids to 30 amino acids in length; (f) 5 amino acids to 25 amino acids in length; or (g) 5 amino acids to 20 amino acids in length.
  • the linker is: (a) 6 amino acids to 50 amino acids in length; (b) 6 amino acids to 45 amino acids in length; (c) 6 amino acids to 40 amino acids in length; (d) 6 amino acids to 35 amino acids in length; (e) 6 amino acids to 30 amino acids in length; (f) 6 amino acids to 25 amino acids in length; or (g) 6 amino acids to 20 amino acids in length.
  • the linker is: (a) 7 amino acids to 40 amino acids in length; (b) 7 amino acids to 35 amino acids in length; (c) 7 amino acids to 30 amino acids in length; (d) 7 amino acids to 25 amino acids in length; (e) 7 amino acids to 20 amino acids in length.
  • the linker is 5 amino acids to 25 amino acids in length. ⁇ ⁇ Attorney Docket No.250298.000954 [00798] In some embodiments, the linker is 10 amino acids to 60 amino acids in length. [00799] In some embodiments, the linker is 20 amino acids to 50 amino acids in length. [00800] In some embodiments, the linker is 25 to 35 amino acids in length. [00801] In some embodiments, the linker is or comprises a multimer of GnS (SEQ ID NO: 239) or SGn (SEQ ID NO: 240), wherein n is an integer from 1 to 10.
  • the linker is or comprises a multimer of G4S (SEQ ID NO: 242).
  • the linker is or comprises two consecutive glycines (2Gly), three consecutive glycines (3Gly), four consecutive glycines (4Gly (SEQ ID NO: 243)), five consecutive glycines (5Gly (SEQ ID NO: 244)), six consecutive glycines (6Gly (SEQ ID NO: 245)), seven consecutive glycines (7Gly (SEQ ID NO: 246)), eight consecutive glycines (8Gly (SEQ ID NO: 247)), or nine consecutive glycines (9Gly (SEQ ID NO: 248)).
  • one or more light chains are universal light chains.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of one or more Fabs are in a CrossmAb arrangement.
  • the multispecific antibody or multispecific antigen-binding comprises an Fc heterodimer.
  • the Fc domains in the Fc heterodimer comprise knob-in- hole mutations as compared to a wild type Fc domain.
  • one Fc domain in the Fc heterodimer comprises amino acid substitutions S354C and T366W (according to EU numbering) as compared to a wild type Fc domain
  • the other Fc domain in the Fc heterodimer comprises amino acid substitutions Y349C, T366S, L368A, and Y407V (according to EU numbering) as compared to a wild type Fc domain.
  • at least one Fc domain in the Fc heterodimer comprises a star mutation as compared to a wild type Fc domain.
  • one Fc domain in the Fc heterodimer comprises amino acid mutations H435R and/or Y436F (according to EU numbering) as compared to a wild type Fc domain.
  • one Fc domain can comprise knob mutation T366W and the other Fc domain can comprise hole mutations T366S, L368A, and Y407V; optionally, one or both Fc domains can comprise star mutations H435R and Y436F.
  • the capsid comprises one or more wild-type AAV capsid polypeptides.
  • the capsid comprises one or more non-wild-type AAV capsid polypeptides. ⁇ ⁇ Attorney Docket No.250298.000954 [00814]
  • the multispecific antibody or multispecific antigen-binding fragment is a bispecific antibody or bispecific antigen-binding fragment thereof.
  • a pharmaceutical composition comprising a multispecific antibody or multispecific antigen-binding fragment described herein, and a pharmaceutically acceptable carrier or excipient.
  • a molecular complex comprising an AAV particle bound to one or more antibodies and/or antigen-binding fragments described herein.
  • said AAV particle comprises one or more mutations in one or more AAV capsid proteins inhibiting the natural tropism of said AAV particle.
  • a pharmaceutical composition comprising a molecular complex described herein and a pharmaceutically acceptable carrier or excipient.
  • a method of preparing the molecular complex described herein comprising incubating said AAV particle in the presence of said one or more antibodies and/or antigen-binding fragments under conditions allowing specific binding of said one or more antibodies and/or antigen-binding fragments to said AAV particle capsid.
  • a molecular complex comprising an AAV particle bound to one or more multispecific antibodies and/or multispecific antigen-binding fragments described herein.
  • said AAV particle comprises one or more mutations in one or more AAV capsid proteins inhibiting the natural tropism of said AAV particle.
  • a pharmaceutical composition comprising a molecular complex described herein and a pharmaceutically acceptable carrier or excipient.
  • a method of preparing a molecular complex described herein comprising incubating said AAV particle in the presence of said one or more multispecific antibodies and/or multispecific antigen-binding fragments under conditions allowing specific binding of said one or more multispecific antibodies and/or multispecific antigen-binding fragments to said AAV particle capsid.
  • a method for targeting an AAV particle to a cell expressing a molecule on the cell surface comprising contacting the cell with a molecular complex described herein or a pharmaceutical composition described herein, wherein said molecular complex comprises one or more multispecific antibodies and/or multispecific antigen-binding fragments which bind to said molecule on the cell surface.
  • a method for delivering a polynucleotide to a cell expressing a molecule on the cell surface comprising contacting the cell with a molecular complex described herein or a pharmaceutical composition described herein, wherein said molecular complex comprises the AAV particle comprising said polynucleotide ⁇ ⁇ Attorney Docket No.250298.000954 and bound to one or more multispecific antibodies and/or multispecific antigen-binding fragments which bind to said molecule on the cell surface.
  • said cell is in a subject and said molecular complex is administered to the subject.
  • said AAV particle does not target said cell in the absence of said one or more multispecific antibodies and/or multispecific antigen-binding fragments.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an scFv comprising a first antigen- binding domain (“ABD1”), the scFv operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the first polypeptide chain can comprise the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTGGGGSGG GGSGGGGSEVQLVESGGGLVQPGNSLTLSCVASGFTFSNYGMHWIRQAPKKGLEWIAMIY ⁇ ⁇ Attorney Docket No.250298.000954 YDSSKMNYADTVKGRFTISRDNSKNTLYLEMNSLRSEDT
  • the first heavy chain region of the first Fab can comprise the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1108), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the scFv can comprise the amino acid sequence EVQLVESGGGLVQPGNSLTLSCVASGFTFSNYGMHWIRQAPKKGLEWIAMIYYDSSKMNY ADTVKGRFTISRDNSKNTLYLEMNSLRSEDTAMYYCAVPTSHYVVDVWGQGVSVTVSSGG GGSGGGGSGGGGSDIQMTQSPASLSASLEEIVTITCQASQDIGNWLAWYQQKPGKSPQLL IYGATSLADGVPSRFSGSRSGTQFSLKISRVQVEDIGIYYCLQAYNTPWTFGGGTKLELK (SEQ ID NO: 1117), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the scFv comprises a first antigen-binding domain (ABD1) comprising or contained within the amino acid sequence EVQLVESGGGLVQPGNSLTLSCVASGFTFSNYGMHWIRQAPKKGLEWIAMIYYDSSKMNY ADTVKGRFTISRDNSKNTLYLEMNSLRSEDTAMYYCAVPTSHYVVDVWGQGVSVTVSS (SEQ ID NO: 1119), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence DIQMTQSPASLSASLEEIVTITCQASQDIGNWLAWYQQKPGKSPQLLIYGATSLADGVPSRF SGSRSGTQFSLKISRVQVEDIGIYYCLQAYNTPWTFGGGTKLELK (SEQ ID NO: 1127), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least
  • the scFv can be operably linked to an Fc domain comprising the amino acid sequence ⁇ ⁇ Attorney Docket No.250298.000954 SKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSPGK (SEQ ID NO: 1137), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second polypeptide chain can comprise the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSR
  • the second heavy chain region of the second Fab can comprise the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1108), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second heavy chain region of the second Fab2 can be operably linked to an Fc domain comprising the amino acid sequence CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK (SEQ ID NO: 1141), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the third polypeptide chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the third polypeptide chain can comprise a first light chain, e.g., that pairs with the first heavy chain region to form Fab1.
  • the first light chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab1 comprises a second antigen binding domain (ABD2) comprising or contained within the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 52), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIK (SEQ ID NO: 10), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least
  • the fourth polypeptide chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the fourth polypeptide chain can comprise a second light chain, e.g., that pairs with the second heavy chain region to form Fab2.
  • the second light chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab2 comprises a third antigen binding domain (ABD3) comprising or contained within the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 52), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, or comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIK (SEQ ID NO: 10), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99%
  • multispecific antibodies can comprise one or more linker, e.g., comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115) and/or the amino acid sequence GGGG (SEQ ID NO: 243).
  • linker e.g., comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115) and/or the amino acid sequence GGGG (SEQ ID NO: 243).
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, the Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); d) a fourth polypeptide chain comprising a second light chain that pairs with the
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • Fab2 is linked to the Fc domain via a linker.
  • the first polypeptide chain can comprise the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSR
  • the first heavy chain region of the first Fab can comprise the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1108), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the Fc domain can comprise the amino acid sequence CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN ⁇ ⁇
  • AKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK (SEQ ID NO: 1141).
  • the second heavy chain region of the second Fab can comprise the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1108), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second polypeptide chain can comprise the amino acid sequence EVQLVESGGGLVQPGNSLTLSCVASGFTFSNYGMHWIRQAPKKGLEWIAMIYYDSSKMNY ADTVKGRFTISRDNSKNTLYLEMNSLRSEDTAMYYCAVPTSHYVVDVWGQGVSVTVSSGG GGSGGGGSGGGGSDIQMTQSPASLSASLEEIVTITCQASQDIGNWLAWYQQKPGKSPQLL IYGATSLADGVPSRFSGSRSGTQFSLKISRVQVEDIGIYYCLQAYNTPWTFGGGTKLELKGG GGSKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK
  • the scFv can comprise the amino acid sequence EVQLVESGGGLVQPGNSLTLSCVASGFTFSNYGMHWIRQAPKKGLEWIAMIYYDSSKMNY ADTVKGRFTISRDNSKNTLYLEMNSLRSEDTAMYYCAVPTSHYVVDVWGQGVSVTVSSGG GGSGGGGSGGGGSDIQMTQSPASLSASLEEIVTITCQASQDIGNWLAWYQQKPGKSPQLL IYGATSLADGVPSRFSGSRSGTQFSLKISRVQVEDIGIYYCLQAYNTPWTFGGGTKLELK (SEQ ID NO: 1117), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the scFv comprises a first antigen-binding domain (ABD1) comprising or contained within the amino acid sequence EVQLVESGGGLVQPGNSLTLSCVASGFTFSNYGMHWIRQAPKKGLEWIAMIYYDSSKMNY ADTVKGRFTISRDNSKNTLYLEMNSLRSEDTAMYYCAVPTSHYVVDVWGQGVSVTVSS (SEQ ID NO: 1119), or a substantially similar sequence thereof having at least 90%, at least ⁇ ⁇ Attorney Docket No.250298.000954 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence DIQMTQSPASLSASLEEIVTITCQASQDIGNWLAWYQQKPGKSPQLLIYGATSLADGVPSRF SGSRSGTQFSLKISRVQVEDIGIYYCLQAYNTPWTFGGGTKLELK (SEQ ID NO: 1127), or a substantially similar sequence thereof having at least
  • the scFv can be operably linked to an Fc domain comprising the amino acid sequence SKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 1137).
  • the third polypeptide chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the third polypeptide chain can comprise a first light chain, e.g., that pairs with the first heavy chain region to form Fab1.
  • the first light chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab1 comprises a second antigen binding domain (ABD2) comprising or contained within the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 52), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF ⁇ ⁇ Attorney Docket No.250298.000954 SGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIK (SEQ ID NO: 10), or a substantially similar sequence thereof having at
  • the fourth polypeptide chain can comprise a second light chain, e.g., that pairs with the second heavy chain region to form Fab2.
  • the second light chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab2 comprises a third antigen binding domain (ABD3) comprising or contained within the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 52), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, or comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIK (SEQ ID NO: 10), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99%
  • multispecific antibodies can comprise one or more linker, e.g., comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115) and/or the amino acid sequence GGGG (SEQ ID NO: 243).
  • linker e.g., comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115) and/or the amino acid sequence GGGG (SEQ ID NO: 243).
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) a first heavy chain region of a first Fab (“Fab1”), the first heavy chain region of Fab1 operably linked to (ii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second scFv comprising a second antigen-binding domain (“ABD2”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”), the second heavy chain region of Fab2 operably linked to (ii) an Fc domain; ⁇ ⁇ Attorney Docket No.250298.000954 c) a third polypeptide chain comprising, in an N- to C-termin
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD3 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD2 binds to a molecule on a cell surface.
  • the first scFv is linked to Fab1 and/or the second scFv is linked to Fab2 via a linker.
  • the first polypeptide chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSSGGGGSGGGGSGGGGSQVQLVESGG
  • the first scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF ⁇ ⁇
  • Attorney Docket No.250298.000954 SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSS (SEQ ID NO: 1155), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the first scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSS (SEQ ID NO: 1358), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the first scFv comprises a first antigen-binding domain (ABD1) comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIK (SEQ ID NO: 1157), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 1159), or a substantially similar sequence thereof having at least 90%, at least 95%, at
  • the first heavy chain region of the first Fab can comprise the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1175), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the first heavy chain region of the first Fab can be operably linked to an Fc domain comprising the amino acid sequence CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 1167).
  • the second polypeptide chain can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSSGGGGSGGGGSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFST YGMHWVRQAPGKGLEWVAVIWHDGSDKY
  • the second scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSS (SEQ ID NO: 1155), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSS (SEQ ID NO: 1358), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second scFv comprises a second antigen-binding domain (ABD2) comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIK (SEQ ID NO: 1157), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at ⁇ ⁇ Attorney Docket No.250298.000954 least 99% sequence identity, and/or comprising or contained within the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 1159), or
  • the second heavy chain region of the second Fab can comprise the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1175), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second heavy chain region of the second Fab can be operably linked to an Fc domain comprising the amino acid sequence CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSPGK (SEQ ID NO: 1167), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the third polypeptide chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the third polypeptide chain can comprise a first light chain, e.g., that pairs with the first heavy chain region to form Fab1.
  • the first light chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ⁇ ⁇ Attorney Docket No.250298.000954 ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab1 comprises a third antigen binding domain (ABD3) comprising or contained within the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS (SEQ ID NO: 795), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIK (SEQ ID NO: 803), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • ABS3
  • the fourth polypeptide chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity [00872]
  • the fourth polypeptide chain can comprise a second light chain, e.g., that pairs with the second heavy chain region to form Fab2.
  • the second light chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab2 comprises a fourth antigen binding domain (ABD4) comprising or contained within the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS (SEQ ID NO: 795), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, or comprising or contained within the ⁇ ⁇ Attorney Docket No.250298.000954 amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIK (SEQ ID NO: 803), or a substantially similar sequence thereof having at least 90%, at least 95%
  • multispecific antibodies or multispecific antigen-binding fragments thereof can comprise one or more of linker, e.g., comprising the amino acid sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 1161) and/or the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115).
  • linker e.g., comprising the amino acid sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 1161) and/or the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115).
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, the Fc domain operably linked to (iii) a first scFv comprising a first antigen-binding domain (“ABD1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain, the Fc domain operably linked to (iii) a second scFv comprising a second antigen-binding domain (“ABD2”); c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD3 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD2 binds to a molecule on a cell surface.
  • the first scFv and/or second scFv is linked to the Fc domain via a linker.
  • the first polypeptide chain can comprise the ⁇ ⁇ Attorney Docket No.250298.000954 amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
  • the first heavy chain region of the first Fab can comprise the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1175), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the first heavy chain region of the first Fab can be operably linked to an Fc domain comprising the amino acid sequence CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 1167), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the first scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID ⁇ ⁇ Attorney Docket No.250298.000954 SWGQGALVTVSS (SEQ ID NO: 1155), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the first scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSS (SEQ ID NO: 1158), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the first scFv comprises a first antigen-binding domain (ABD1) comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIK (SEQ ID NO: 1157), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 1159), or a substantially similar sequence thereof having at least 90%, at least 95%, at
  • the second polypeptide chain can comprise the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYGSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
  • the second heavy chain region of the second Fab can comprise the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT (SEQ ID NO: 1175), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second heavy chain region of the second Fab can be operably linked to an Fc domain comprising the amino acid sequence CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYGSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSPGK (SEQ ID NO: 1167), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSS (SEQ ID NO: 1155), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second scFv can comprise the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIKGGGGSGGGGSGGGG SGGGGSQVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKGLEWIGHIH YSERTSHNPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFID SWGQGALVTVSS (SEQ ID NO: 1358), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the second scFv comprises a second antigen-binding domain (ABD2) comprising or contained within the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRF SGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGCGTRLEIK (SEQ ID NO: 1157), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence ⁇ ⁇ Attorney Docket No.250298.000954 QVHLQESGPGLVMPSQILSLSCVISGDSIRNGGYYWTWTRQQPGKCLEWIGHIHYSERTSH NPSLQSRVIMSIDTSENKFSLKLTSVTVADTAIYYCARGRDTMVRGSITTSGHFIDSWGQGA LVTVSS (SEQ ID NO: 1159), or
  • the third polypeptide chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the third polypeptide chain can comprise a first light chain, e.g., that pairs with the first heavy chain region to form Fab1.
  • the first light chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab1 comprises a third antigen binding domain (ABD3) comprising or contained within the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS (SEQ ID NO: 795), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, and/or comprising or contained within the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIK (SEQ ID NO: 803), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • ABS3
  • the fourth polypeptide chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR ⁇ ⁇ Attorney Docket No.250298.000954 FSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • the fourth polypeptide chain can comprise a second light chain, e.g., that pairs with the second heavy chain region to form Fab2.
  • the second light chain can comprise the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 813), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • Fab2 comprises a fourth antigen binding domain (ABD4) comprising or contained within the amino acid sequence QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWHDGSDK YYVDSVKGRFSIARDNSKNTLYLQMNSLRVEDTGIYYCARRGIRGTVFDHWGLGTLVTVSS (SEQ ID NO: 795), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity, or comprising or contained within the amino acid sequence DIQMTQSPSTLSASVGDRVTLTCRASQSISNKLAWYQQKPGKAPNLLIYKASNLESGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSWTFGQGTKVEIK (SEQ ID NO: 803), or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.
  • ABS4 fourth anti
  • the anti-AAV x anti-mTfR antibodies, the anti-AAV x anti- ⁇ CACNG1 antibodies, or the anti-AAV x anti-ASGR antibodies, or antigen-binding fragments thereof, described herein comprise the amino acid sequence set forth in SEQ ID NOs: 1106, 1139, 20, 1149, 1151, 1153, 1173, 1169, 813, 1228, 1232, 1234, 1236, 1237, 1238, 1328, 1330, 1332, 1341, 1506, 1348, 1354, 1357, 1361, 1362, 1369, 1373, 1342, 1375, 1379, 1381, 1384, 1387, 1390, 1393, 1395, 1502, 1504, 1508, or 1510, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NOs:
  • the nucleotide sequence that encodes the anti-AAV x anti-mTfR antibodies, the anti-AAV x anti- ⁇ CACNG1 ⁇ ⁇ Attorney Docket No.250298.000954 antibodies, or the anti-AAV x anti-ASGR antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence that encodes the amino acid sequence of SEQ ID NOs: 1106, 1139, 20, 1149, 1151, 1153, 1173, 1169, 813, 1228, 1232, 1234, 1236, 1237, 1238, 1328, 1330, 1332, 1341, 1506, 1348, 1354, 1357, 1361, 1362, 1369, 1373, 1342, 1375, 1379, 1381, 1384, 1387, 1390, 1393, 1395, 1502, 1504, 1508, or 1510, or a variant thereof having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%
  • the nucleotide sequence that encodes the anti-AAV x anti-mTfR antibodies, the anti-AAV x anti- ⁇ CACNG1 antibodies, or the anti-AAV x anti-ASGR antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 19, 1105, 1138, 1145, 1500, 1148, 1498, 1150, 1499, 1152, 1172, 1168, 1505, 812, 1240, 1258, 1265, 1269, 1287, 1298, 1316, 1329, 1331, 1333, 1340, 1346, 1347, 1353, 1356, 1360, 1368, 1372, 1374, 1378, 1380, 1383, 1386, 1389, 1392, 1394, 1501, 1503, 1507, or 1509, or a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 9
  • the anti-AAV x anti-mTfR antibodies, the anti-AAV x anti- ⁇ CACNG1 antibodies, or the anti-AAV x anti-ASGR antibodies, or antigen-binding fragments thereof comprise the amino acid sequence set forth in SEQ ID NOs: 1106, 1139, 20, 1149, 1151, 1153, 1173, 1169, 813, 1228, 1232, 1234, 1236, 1237, 1238, 1328, 1330, 1332, 1341, 1506, 1348, 1354, 1357, 1361, 1362, 1369, 1373, 1342, 1375, 1379, 1381, 1384, 1387, 1390, 1393, 1395, 1502, 1504, 1508, or 1510.
  • the nucleotide sequence that encodes the anti-AAV x anti-mTfR antibodies, the anti-AAV x anti- ⁇ CACNG1 antibodies, or the anti-AAV x anti-ASGR antibodies, or antigen-binding fragments thereof comprises the nucleotide sequence set forth in SEQ ID NOs: 19, 1105, 1138, 1145, 1500, 1148, 1498, 1150, 1499, 1152, 1172, 1168, 1505, 812, 1240, 1258, 1265, 1269, 1287, 1298, 1316, 1329, 1331, 1333, 1340, 1346, 1347, 1353, 1356, 1360, 1368, 1372, 1374, 1378, 1380, 1383, 1386, 1389, 1392, 1394, 1501, 1503, 1507, or 1509.
  • multispecific antibodies or multispecific antigen-binding fragments thereof can comprise one or more of linker, e.g., comprising the amino acid sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 1161) ⁇ ⁇ Attorney Docket No.250298.000954 and/or the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115).
  • linker e.g., comprising the amino acid sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 1161) ⁇ ⁇ Attorney Docket No.250298.000954 and/or the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 1115).
  • the epitope on a capsid of an AAV and/or molecule on a cell surface to which the antigen-binding molecules of the present disclosure bind may consist of a single contiguous sequence of 3 or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) amino acids of an AAV capsid and/or molecule on a cell surface protein.
  • the epitope may consist of a plurality of non-contiguous amino acids (or amino acid sequences).
  • epitope can refer to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule known as a paratope.
  • a single antigen may have more than one epitope.
  • different antibodies may bind to different areas on an antigen and may have different biological effects.
  • Epitopes may be either conformational or linear.
  • a conformational epitope is produced by spatially juxtaposed amino acids from different segments of the linear polypeptide chain.
  • a linear epitope is one produced by adjacent amino acid residues in a polypeptide chain.
  • an epitope may include moieties of saccharides, phosphoryl groups, or sulfonyl groups on the antigen.
  • Various techniques known to persons of ordinary skill in the art can be used to determine whether an antigen-binding domain of an antibody “interacts with one or more amino acids” within a polypeptide or protein.
  • Exemplary techniques include, e.g., cryogenic electron microscopy (cryo-EM), routine cross-blocking assay such as that described in Antibodies, Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harb., NY), alanine scanning mutational analysis, peptide blots analysis (Reineke, 2004, Methods Mol Biol 248:443-463), and peptide cleavage analysis.
  • the hydrogen/deuterium exchange method involves deuterium-labeling the protein of interest, followed by binding the antibody to the deuterium-labeled protein. Next, the protein/antibody complex is transferred to water to allow hydrogen-deuterium exchange to occur at all residues except for the residues protected by the antibody (which remain deuterium- labeled).
  • the target protein After dissociation of the antibody, the target protein is subjected to protease cleavage and mass spectrometry analysis, thereby revealing the deuterium-labeled residues which correspond to the specific amino acids with which the antibody interacts. See, e.g., ⁇ ⁇ Attorney Docket No.250298.000954 Ehring (1999) Analytical Biochemistry 267(2):252-259; Engen and Smith (2001) Anal. Chem. 73:256A-265A. X-ray crystallography of the antigen/antibody complex may also be used for epitope mapping purposes.
  • anti-AAV antibodies that bind to the same epitope as any of the specific exemplary antibodies described herein (e.g., antibodies comprising any of the amino acid sequences as set forth in Table 1 herein).
  • the present disclosure also includes anti-AAV antibodies that compete for binding to a capsid of an AAV particle with any of the specific exemplary antibodies described herein (e.g., antibodies comprising any of the amino acid sequences as set forth in Table 1 herein).
  • anti-cell surface molecule antibodies that bind to the same epitope as any of the specific exemplary antibodies described herein (e.g., antibodies comprising any of the amino acid sequences as set forth in Tables 5, 7, and 10 herein).
  • the present disclosure also includes anti-cell surface molecule antibodies that compete for binding to a cell surface molecule with any of the specific exemplary antibodies described herein (e.g., antibodies comprising any of the amino acid sequences as set forth in Tables 5, 7, and 10 herein).
  • bispecific antigen-binding molecules comprising a first antigen-binding domain that specifically binds to a capsid of an AAV, and a second antigen-binding domain that specifically binds to a molecule on a cell surface, wherein the first antigen-binding domain competes for binding to a capsid of an AAV with any of the specific exemplary AAV-specific antigen-binding domains described herein, and/or wherein the second antigen-binding domain competes for binding to a molecule on a surface of a cell with any of the specific exemplary cell surface-specific antigen-binding domains described herein.
  • One can easily determine whether a particular antigen-binding molecule e.g., antibody
  • a particular antigen-binding molecule e.g., antibody
  • antigen-binding domain thereof binds to the same epitope as, or competes for binding with, a reference antigen-binding molecule of the present disclosure by using routine methods known in the art. For example, to determine if a test antibody binds to the same epitope on an AAV capsid (or cell surface molecule) protein as a reference bispecific antigen-binding molecule of the present disclosure, the reference bispecific molecule is first allowed to bind to the AAV capsid (or cell surface molecule) protein . Next, the ability of a test antibody to bind to the AAV capsid (or cell surface molecule) protein is assessed.
  • test antibody If the test antibody is able to bind to the AAV capsid (or cell surface molecule) protein following saturation binding with the reference bispecific antigen-binding molecule, it can be concluded that the test antibody binds to a different epitope of the AAV capsid protein (or cell surface molecule) than the reference bispecific antigen-binding molecule.
  • test antibody may bind to the same epitope of the AAV capsid protein as the epitope bound by the reference bispecific antigen-binding molecule of the disclosure. Additional routine experimentation (e.g., peptide mutation and binding analyses) can then be carried out to confirm whether the observed lack of binding of the test antibody is in fact due to binding to the same epitope as the reference bispecific antigen-binding molecule or if steric blocking (or another phenomenon) is responsible for the lack of observed binding.
  • two antigen-binding proteins bind to the same (or overlapping) epitope if, e.g., a 1-, 5-, 10-, 20- or 100-fold excess of one antigen-binding protein inhibits binding of the other by at least 50% but preferably 75%, 90%, or even 99% as measured in a competitive binding assay (see, e.g., Junghans et al., Cancer Res. 1990:50:1495-1502).
  • two antigen-binding proteins are deemed to bind to the same epitope if essentially all amino acid mutations in the antigen that reduce or eliminate binding of one antigen-binding protein reduce or eliminate binding of the other.
  • Two antigen- binding proteins are deemed to have “overlapping epitopes” if only a subset of the amino acid mutations that reduce or eliminate binding of one antigen-binding protein reduce or eliminate binding of the other.
  • the above-described binding methodology is performed in two orientations: As an example, in a first orientation, the reference antigen- binding molecule is allowed to bind to an AAV capsid (or cell surface molecule) protein under saturating conditions followed by assessment of binding of the test antibody to AAV capsid (or cell surface molecule) protein. In a second orientation, the test antibody is allowed to bind to a AAV capsid (or cell surface molecule) protein under saturating conditions followed by assessment of binding of the reference antigen-binding molecule to the AAV capsid (or cell surface molecule) protein.
  • test antibody and the reference antigen-binding molecule compete for binding to the AAV capsid (or cell surface molecule) protein.
  • an antibody that competes for binding with a reference antigen-binding molecule may not necessarily bind to the same epitope as the reference antibody but may sterically block binding of the reference antibody by binding an overlapping or adjacent epitope.
  • Antigen-binding domains specific for particular antigens can be prepared by any antibody generating technology known in the art. Once obtained, two different antigen- binding domains, specific for two different antigens, can be appropriately arranged relative to one another to produce a bispecific antigen-binding molecule of the present disclosure using routine methods. (A discussion of exemplary bispecific antibody formats that can be used to construct the bispecific antigen-binding molecules of the present disclosure is provided elsewhere herein).
  • one or more of the individual components (e.g., heavy and light chains) of the multispecific antigen-binding molecules of the disclosure are derived from chimeric, humanized, or fully human antibodies. Methods for making such antibodies are well known in the art.
  • one or more of the heavy and/or light chains of the bispecific antigen-binding molecules of the present disclosure can be prepared using VELOCIMMUNETM technology. Using VELOCIMMUNETM technology (or any other human antibody generating technology), high affinity chimeric antibodies to a particular antigen are initially isolated having a human variable region and a mouse constant region. The antibodies are characterized and selected for desirable characteristics, including affinity, selectivity, epitope, etc.
  • mice constant regions are replaced with a desired human constant region to generate fully human heavy and/or light chains that can be incorporated into the bispecific antigen-binding molecules of the present disclosure.
  • Genetically engineered animals may be used to make human bispecific antigen- binding molecules.
  • a genetically modified mouse can be used which is incapable of rearranging and expressing an endogenous mouse immunoglobulin light chain variable sequence, wherein the mouse expresses only one or two human light chain variable domains encoded by human immunoglobulin sequences operably linked to the mouse kappa constant gene at the endogenous mouse kappa locus.
  • Fully human can refer to an antibody, or antigen-binding fragment or immunoglobulin domain thereof, comprising an amino acid sequence encoded by a DNA derived from a human sequence over the entire length of each polypeptide of the antibody or antigen-binding fragment or immunoglobulin domain thereof.
  • the fully human sequence is derived from a protein endogenous to a human.
  • the fully human protein or protein sequence comprises a chimeric sequence wherein each component sequence is derived from human sequence. While not being bound by any one theory, chimeric proteins or chimeric sequences are generally designed to ⁇ ⁇ Attorney Docket No.250298.000954 minimize the creation of immunogenic epitopes in the junctions of component sequences, e.g., compared to any wild-type human immunoglobulin regions or domains. Bioequivalents [00901]
  • the present disclosure encompasses antigen-binding molecules having amino acid sequences that vary from those of the exemplary molecules disclosed herein but that retain the ability to bind to a capsid of an AAV particle and/or a molecule on a surface of a cell.
  • Such variant molecules may comprise one or more additions, deletions, or substitutions of amino acids when compared to parent sequence but exhibit biological activity that is essentially equivalent to that of the described bispecific antigen-binding molecules.
  • the present disclosure includes antigen-binding molecules that are bioequivalent to any of the exemplary antigen-binding molecules set forth herein. Two antigen-binding proteins, or antibodies, are considered bioequivalent if, for example, they are pharmaceutical equivalents or pharmaceutical alternatives whose rate and extent of absorption do not show a significant difference when administered at the same molar dose under similar experimental conditions, either single does or multiple dose.
  • antigen-binding proteins will be considered equivalents or pharmaceutical alternatives if they are equivalent in the extent of their absorption but not in their rate of absorption and yet may be considered bioequivalent because such differences in the rate of absorption are intentional and are reflected in the labeling, are not essential to the attainment of effective body drug concentrations on, e.g., chronic use, and are considered medically insignificant for the particular drug product studied.
  • two antigen-binding proteins are bioequivalent if there are no clinically meaningful differences in their safety, purity, and potency.
  • two antigen-binding proteins are bioequivalent if a patient can be switched one or more times between the reference product and the biological product without an expected increase in the risk of adverse effects, including a clinically significant change in immunogenicity, or diminished effectiveness, as compared to continued therapy without such switching.
  • two antigen-binding proteins are bioequivalent if they both act by a common mechanism or mechanisms of action for the condition or conditions of use, to the extent that such mechanisms are known.
  • Bioequivalence may be demonstrated by in vivo and in vitro methods.
  • Bioequivalence measures include, e.g., (a) an in vivo test in humans or other mammals, in which the concentration of the antibody or its metabolites is measured in blood, plasma, serum, or other biological fluid as a function of time; (b) an in vitro test that has been ⁇ ⁇ Attorney Docket No.250298.000954 correlated with and is reasonably predictive of human in vivo bioavailability data; (c) an in vivo test in humans or other mammals in which the appropriate acute pharmacological effect of the antibody (or its target) is measured as a function of time; and (d) in a well-controlled clinical trial that establishes safety, efficacy, or bioavailability or bioequivalence of an antigen-binding protein.
  • Bioequivalent variants of the exemplary bispecific antigen-binding molecules set forth herein may be constructed by, for example, making various substitutions of residues or sequences or deleting terminal or internal residues or sequences not needed for biological activity.
  • cysteine residues not essential for biological activity can be deleted or replaced with other amino acids to prevent formation of unnecessary or incorrect intramolecular disulfide bridges upon renaturation.
  • bioequivalent antigen- binding proteins may include variants of the exemplary bispecific antigen-binding molecules set forth herein comprising amino acid changes which modify the glycosylation characteristics of the molecules, e.g., mutations which eliminate or remove glycosylation.
  • the disclosure also provides nucleic acids encoding the multispecific binding molecules of the disclosure.
  • the multispecific binding molecules are encoded by a single nucleic acid.
  • the multispecific binding molecules can be encoded by a plurality (e.g., two, three, four, or more) nucleic acids.
  • a single nucleic acid can encode a multispecific binding molecule that comprises a single polypeptide chain, a multispecific binding molecule that comprises two or more polypeptide chains, or a portion of a multispecific binding molecule that comprises more than two polypeptide chains (for example, a single nucleic acid can encode two polypeptide chains of a multispecific binding molecule comprising three, four, or more polypeptide chains, or three polypeptide chains of a multispecific binding molecule comprising four or more polypeptide chains).
  • the open reading frames encoding two or more polypeptide chains can be under the control of separate transcriptional regulatory elements (e.g., promoters and/or enhancers).
  • a multispecific binding molecule comprising two or more polypeptide chains is encoded by two or more nucleic acids.
  • the number of nucleic acids ⁇ ⁇ Attorney Docket No.250298.000954 encoding a multispecific binding molecule can be equal to or less than the number of polypeptide chains in the multispecific binding molecule (for example, when more than one polypeptide chains are encoded by a single nucleic acid).
  • the nucleic acids of the disclosure can be DNA (e.g., plasmid) or RNA (e.g., mRNA).
  • Cells [00912]
  • the disclosure also provides host cells comprising a nucleic acid of the disclosure.
  • the host cells are genetically engineered to comprise one or more nucleic acids described herein.
  • the host cells are genetically engineered by using an expression cassette.
  • expression cassette refers to nucleotide sequences, which are capable of affecting expression of a gene in hosts compatible with such sequences. Such cassettes may include a promoter, an open reading frame with or without introns, and a termination signal.
  • the disclosure also provides host cells comprising the vectors described herein.
  • the cell can be, but is not limited to, a eukaryotic cell, a bacterial cell, an insect cell, or a human cell.
  • Suitable eukaryotic cells include, but are not limited to, Vero cells, HeLa cells, COS cells, CHO cells, HEK293 cells, BHK cells, and MDCKII cells.
  • Suitable insect cells include, but are not limited to, Sf9 cells.
  • the present disclosure provides a pharmaceutical composition comprising an antigen-binding molecule (e.g., anti-AAV monospecific antibody or anti-AAV x anti-cell surface molecule multispecific antibody) described herein and/or at least one viral capsid protein (e.g., a capsid protein of an AAV particle) described herein.
  • an antigen-binding molecule e.g., anti-AAV monospecific antibody or anti-AAV x anti-cell surface molecule multispecific antibody
  • viral capsid protein e.g., a capsid protein of an AAV particle
  • pharmaceutical compositions comprising a viral capsid protein e.g., an AAV particle
  • the pharmaceutical compositions of the disclosure are formulated with suitable carriers, excipients, and other agents that provide improved transfer, delivery, tolerance, and the like.
  • formulations can be found in the formulary known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic) containing vesicles (such as LIPOFECTINTM, Life Technologies, Carlsbad, CA), DNA conjugates, anhydrous absorption ⁇ ⁇ Attorney Docket No.250298.000954 pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax.
  • vesicles such as LIPOFECTINTM, Life Technologies, Carlsbad, CA
  • the dose of antigen-binding molecule administered to a patient may vary depending upon the age and the size of the patient, target disease, conditions, route of administration, and the like.
  • the preferred dose is typically calculated according to body weight or body surface area.
  • bispecific antigen-binding molecule of the present disclosure When a bispecific antigen-binding molecule of the present disclosure is used for therapeutic purposes in an adult patient, it may be advantageous to intravenously administer the bispecific antigen-binding molecule of the present disclosure normally at a single dose of about 0.01 to about 20 mg/kg body weight, more preferably about 0.02 to about 7, about 0.03 to about 5, or about 0.05 to about 3 mg/kg body weight. Depending on the severity of the condition, the frequency and the duration of the treatment can be adjusted. Effective dosages and schedules for administering a bispecific antigen- binding molecule may be determined empirically; for example, patient progress can be monitored by periodic assessment, and the dose adjusted accordingly.
  • Dose ranges and frequency of administration of a viral vector described herein can vary depending on the nature of, e.g., the AAV, and the medical condition, as well as parameters of a specific patient and the route of administration used.
  • viral vector compositions can be administered to a subject at a dose ranging from about 1 ⁇ 10 5 plaque forming units (pfu) to about 1 ⁇ 10 15 pfu, depending on mode of administration, the route of administration, the nature of the disease and condition of the subject.
  • the viral vector compositions can be administered at a dose ranging from about 1 ⁇ 10 8 pfu to about 1 ⁇ 10 15 pfu, or from about 1 ⁇ 10 10 pfu to about 1 ⁇ 10 15 pfu, or from about 1 ⁇ 10 8 pfu to about 1 ⁇ 10 12 pfu.
  • a more accurate dose can also depend on the subject in which it is being administered. For example, a lower dose may be required if the subject is juvenile, and a higher dose may be required if the subject is an adult human subject. In certain embodiments, a more accurate dose can depend on the weight of the subject.
  • a juvenile human subject can receive from about 1 ⁇ 10 8 pfu to about 1 ⁇ 10 10 pfu, while an adult human subject can receive a dose from about 1 ⁇ 10 10 pfu to about 1 ⁇ 10 12 pfu.
  • the dose of the multispecific antibodies or multispecific antigen-binding fragments (or pharmaceutical compositions thereof) administered to the subject is between about 1x10 5 vector genomes ⁇ ⁇ Attorney Docket No.250298.000954 (vg) to about 1x10 16 vg.
  • the dose of the multispecific antibodies or multispecific antigen-binding fragments administered to the subject is between about 1x10 6 vg to about 1x10 9 vg, about 1x10 7 vg to about 1x10 10 vg, about 1x10 8 vg to about 1x10 11 vg, about 1x10 9 vg to about 1x10 12 vg, about 1x10 10 vg to about 1x10 13 vg, about 1x10 11 vg to about 1x10 14 vg, about 1x10 12 vg to about 1x10 15 vg, about 1x10 13 vg to about 1x10 16 vg, or about 1x10 14 vg to about 1x10 16 vg.
  • the dose of the multispecific antibodies or multispecific antigen-binding fragments administered to the subject is between about 1x10 10 vg to about 1x10 16 vg. In certain embodiments, for the methods disclosed herein, the dose of the multispecific antibodies or multispecific antigen-binding fragments administered to the subject is at least about 1x10 6 vg, at least about 1x10 7 vg, at least about 1x10 8 vg, at least about 1x10 9 vg, at least about 1x10 10 vg, at least about 1x10 11 vg, at least about 1x10 12 vg, at least about 1x10 12 vg, at least about 1x10 13 vg, at least about 1x10 14 vg, or at least about 1x10 15 vg.
  • the vg is total vector genome per subject.
  • the dose of the multispecific antibodies or multispecific antigen-binding fragments administered to the subject is about 1x10 6 vg, about 1x10 7 vg, about 1x10 8 vg, about 2x10 8 vg, about 3x10 8 vg, about 4x10 8 vg, about 5x10 8 vg, about 6x10 8 vg, about 7x10 8 vg, about 8x10 8 vg, about 9x10 8 vg, about 1x10 9 vg, about 2x10 9 vg, about 3x10 9 vg, about 4x10 9 vg, about 5x10 9 vg, about 6x10 9 vg, about 7x10 9 vg, about 8x10 9 vg, about 9x10 9 vg, about 1x10 10 vg, about 2x10 10 vg, about 3x10 10 v
  • the vg is total vector genome per subject.
  • the dose of the multispecific antibodies or multispecific antigen-binding fragments (or pharmaceutical compositions thereof) administered to the subject is about 1x10 12 , 1x10 13 , 1x10 14 , 1x10 15 , and 1x10 16 vg/mL.
  • doses of multispecific antibodies or multispecific antigen-binding fragments include about 1x10 12 , about 1x10 13 , about 1x10 14 , about 1x10 15 , and about 1x10 16 vg/mL, or between about 1x10 12 to about 1x10 16 , between about 1x10 12 to about 1x10 15 , between about 1x10 12 to about 1x10 14 , between about 1x10 12 to about 1x10 13 , between about 1x10 13 to about 1x10 16 , between about 1x10 14 to about 1x10 16 , between ⁇ ⁇ Attorney Docket No.250298.000954 about 1x10 15 to about 1x10 16 , or between about 1x10 13 to about 1x10 15 vg/mL.
  • an AAV is dosed or incubated in combination with the multispecific antibodies or multispecific antigen-binding fragments (or pharmaceutical compositions thereof) described herein.
  • the AAV viral genome to the antibody molar ratio e.g., the VG:Ab ratio or AAV:Ab ratio
  • the AAV viral genome to the antibody molar ratio is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:31, 1:32, 1:33, 1:34, 1:35, 1:36, 1:37, 1:38, 1:39, 1:40, 1:41, 1:42, 1:43, 1:44, 1:45, 1:46, 1:47, 1:48, 1:49, 1:50, 1:51,
  • the AAV viral genome to the antibody molar ratio (e.g., the VG:Ab ratio or AAV:Ab ratio) is 1:1, 1:3, 1:9, 1:27, 1:81, 1:243, 1:729, or 1:2187.
  • the AAV viral genome to the antibody molar ratio is about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, about 1:30, about 1:31, about 1:32, about 1:33, about 1:34, about 1:35, about 1:36, about 1:37, about 1:38, about 1:39, about 1:40, about 1:41, about 1:42, about 1:43, about 1:44, about 1:45, about 1:46, about 1:47, about 1:48, about 1:49, about 1:50, about 1:51, about 1:52
  • the AAV viral genome to the antibody molar ratio (e.g., the VG:Ab ratio or AAV:Ab ratio) is about 1:1, about 1:3, about 1:9, about 1:27, about 1:81, about 1:243, about 1:729, or about 1:2187.
  • the AAV viral genome to the antibody molar ratio is less than about 1:1, about 1:1 to about 1:2, about 1:2 to ⁇ ⁇ Attorney Docket No.250298.000954 about 1:3, about 1:3 to about 1:4, about 1:4 to about 1:5, about 1:5 to about 1:6, about 1:6 to about 1:7, about 1:7 to about 1:8, about 1:8 to about 1:9, about 1:9 to about 1:10, about 1:10 to about 1:11, about 1:11 to about 1:12, about 1:12 to about 1:13, about 1:13 to about 1:14, about 1:14 to about 1:15, about 1:15 to about 1:16, about 1:16 to about 1:17, about 1:17 to about 1:18, about 1:18 to about 1:19, about 1:19 to about 1:20, about 1:20 to about 1:21, about 1:21 to about 1:22, about 1:22 to about 1:23, about 1:23 to about 1:24, about 1:24 to about 1:25, about 1:25 to about 1:26, about
  • the AAV viral genome to the antibody molar ratio (e.g., the VG:Ab ratio or AAV:Ab ratio) is less than about 1:1, about 1:1 to about 1:3, about 1:3 to about 1:9, about 1:9 to about 1:27, about 1:27 to about 1:81, about 1:81 to about 1:243, about 1:243 to about 1:729, about 1:729 to about 1:2187, or at least about 1:2187.
  • Various delivery systems are known and can be used to administer the pharmaceutical composition of the disclosure, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the mutant viruses, receptor mediated endocytosis (see, e.g., Wu et al., 1987, J. Biol. Chem.262:4429-4432).
  • Methods of introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes.
  • compositions may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration can be systemic or local.
  • a pharmaceutical composition of the present disclosure can be delivered subcutaneously or intravenously with a standard needle and syringe.
  • a pen delivery device readily has applications in delivering a pharmaceutical composition of the present disclosure.
  • Such a pen delivery device can be reusable or disposable.
  • a reusable pen delivery device generally utilizes a replaceable cartridge that contains a pharmaceutical composition.
  • the empty cartridge can readily be discarded and replaced with a new cartridge that contains the pharmaceutical composition.
  • the pen delivery device can then be reused.
  • a disposable pen delivery device there is no replaceable cartridge. Rather, the disposable pen delivery device comes prefilled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is ⁇ ⁇ Attorney Docket No.250298.000954 discarded.
  • Numerous reusable pen and autoinjector delivery devices have applications in the subcutaneous delivery of a pharmaceutical composition of the present disclosure.
  • Examples include, but are not limited to AUTOPENTM (Owen Mumford, Inc., Woodstock, UK), DISETRONICTM pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75/25TM pen, HUMALOGTM pen, HUMALIN 70/30TM pen (Eli Lilly and Co., Indianapolis, IN), NOVOPENTM I, II, and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIORTM (Novo Nordisk, Copenhagen, Denmark), BDTM pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPENTM, OPTIPEN PROTM, OPTIPEN STARLETTM, and OPTICLIKTM (sanofi-aventis, Frankfurt, Germany), to name only a few.
  • Examples of disposable pen delivery devices having applications in subcutaneous delivery of a pharmaceutical composition of the present disclosure include, but are not limited to the SOLOSTARTM pen (sanofi-aventis), the FLEXPENTM (Novo Nordisk), and the KWIKPENTM (Eli Lilly), the SURECLICK TM Autoinjector (Amgen, Thousand Oaks, CA), the PENLET TM (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, L.P.), and the HUMIRA TM Pen (Abbott Labs, Abbott Park IL), to name only a few.
  • the pharmaceutical composition can be delivered in a controlled release system.
  • a pump may be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng.14:201).
  • polymeric ⁇ eds.
  • a controlled release system can be placed in proximity of the composition’s target, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, 1984, in Medical Applications of Controlled Release, supra, vol.2, pp.115-138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.
  • the injectable preparations may include dosage forms for intravenous, subcutaneous, intracutaneous, and intramuscular injections, drip infusions, etc. These injectable preparations may be prepared by methods publicly known. For example, the injectable preparations may be prepared, e.g., by dissolving, suspending, or emulsifying the antibody or its salt described above in a sterile aqueous medium or an oily medium conventionally used for injections.
  • aqueous medium for injections there are, for example, physiological saline, an isotonic solution containing glucose and other auxiliary agents, etc., which may be used in combination with an appropriate solubilizing agent such as an alcohol (e.g., ethanol), a polyalcohol (e.g., propylene glycol, polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc.
  • an alcohol e.g., ethanol
  • a polyalcohol e.g., propylene glycol, polyethylene glycol
  • a nonionic surfactant e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil
  • the oily medium there are employed, e.g., sesame oil, ⁇ ⁇ Attorney Docket No.250298.000954 soybean oil, etc., which may be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc.
  • a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc.
  • the injection thus prepared is preferably filled in an appropriate ampoule.
  • the pharmaceutical compositions for oral or parenteral use described above are prepared into dosage forms in a unit dose suited to fit a dose of the active ingredients.
  • dosage forms in a unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc.
  • the amount of the aforesaid antibody contained is generally about 5 to about 500 mg per dosage form in a unit dose; especially in the form of injection, it is preferred that the aforesaid antibody is contained in about 5 to about 100 mg and in about 10 to about 250 mg for the other dosage forms.
  • Uses of the Antigen-Binding Molecules [00930] The present disclosure includes methods of using the any of antigen-binding molecules as disclosed herein or a pharmaceutically acceptable carrier or diluent.
  • a method for reducing off-target effects of an AAV particle in vivo comprising administering said AAV particle in a molecular complex comprising said AAV particle bound to one or more antibodies and/or antigen-binding fragments described herein or one or more multispecific antibodies and/or multispecific antigen-binding fragments described herein.
  • a method for enhancing target specificity of an AAV particle in vivo comprising administering said AAV particle in a molecular complex comprising said AAV particle bound to one or more antibodies and/or antigen-binding fragments described herein or one or more multispecific antibodies and/or multispecific antigen-binding fragments described herein.
  • provided herein is a method for determining the presence and/or identity of an AAV capsid in a sample, comprising contacting the sample with the antibody or antigen-binding fragment described herein, wherein said antibody or antigen-binding fragment binds to said AAV particle.
  • a method for visualizing an AAV particle in a subject comprising administering to the subject the antibody or antigen-binding fragment described herein, wherein said antibody or antigen-binding fragment binds to said AAV particle and is attached to a detectable agent.
  • a method for assessing quality of an AAV particle preparation comprising contacting said AAV particle preparation with the antibody or antigen-binding fragment described herein , wherein said antibody or antigen-binding fragment binds to said AAV particle and is attached to a detectable agent.
  • ⁇ Attorney Docket No.250298.000954
  • an affinity matrix comprising the antibody or antigen-binding fragment described herein or the multispecific antibody or multispecific antigen-binding fragment described herein attached to a solid support.
  • provided herein is a method of isolating or purifying an AAV capsid or an AAV particle from a sample, comprising contacting the sample with the affinity matrix described herein.
  • a method for inhibiting an infection or transduction of a cell mediated by an AAV particle comprising contacting said AAV particle with said cell in the presence of the antibody or antigen-binding fragment described herein.
  • the cell is in a subject and the antibody or antigen-binding fragment is administered to said subject.
  • a method for neutralizing a capsid of an AAV particle comprising contacting said AAV particle with the antibody or antigen-binding fragment described herein.
  • the method for neutralizing a capsid achieves detargeting of the AAV particle from a cell and /or tissue-type naturally targeted by the capsid in the absence of the antibody of antigen-binding fragment.
  • the methods comprise administering to a subject in need thereof a therapeutic composition comprising any of the antibodies or antigen-binding molecules as disclosed herein and a pharmaceutically acceptable carrier or diluent.
  • an antigen-binding molecule described herein may be administered to a subject separately from an AAV particle, or in a pre-complexed form with an AAV particle.
  • an antigen- binding molecule and an AAV particle may be administered as a molecular complex described herein.
  • an antigen-binding molecule and an AAV particle when administered separately, the antigen-binding molecule may be administered at the same time as the AAV particle.
  • an antigen-binding molecule and an AAV particle may be administered separately over a defined time course.
  • multiple doses of an antigen-binding molecule and/or an AAV particle described herein may be ⁇ ⁇ Attorney Docket No.250298.000954 administered to a subject over a defined time course.
  • the methods according to such aspects of the disclosure may comprise sequentially administering to a subject multiple doses of an antigen-binding molecule and/or an AAV particle of the disclosure.
  • sequentially administering means that each dose of an antigen-binding molecule and/or AAV is administered to the subject at a different point in time, e.g., on different days separated by a predetermined interval (e.g., hours, days, weeks, or months).
  • the present disclosure includes methods which comprise sequentially administering to the patient a single initial dose of an antigen-binding molecule and/or AAV, followed by one or more secondary doses of the antigen-binding molecule and/or AAV, and optionally followed by one or more tertiary doses of the antigen-binding molecule and/or AAV.
  • the terms “initial dose,” “secondary doses,” and “tertiary doses,” can refer to the temporal sequence of administration of the antigen-binding molecule and/or AAV of the disclosure.
  • the “initial dose” is the dose which is administered at the beginning of the treatment regimen (also referred to as the “baseline dose”);
  • the “secondary doses” are the doses which are administered after the initial dose;
  • the “tertiary doses” are the doses which are administered after the secondary doses.
  • the initial, secondary, and tertiary doses may all contain the same amount of the antigen-binding molecule and/or AAV, but generally may differ from one another in terms of frequency of administration.
  • the amount of an antigen-binding molecule and/or AAV contained in the initial, secondary, and/or tertiary doses varies from one another (e.g., adjusted up or down as appropriate) during the course of treatment.
  • two or more (e.g., 2, 3, 4, or 5) doses are administered at the beginning of the treatment regimen as “loading doses” followed by subsequent doses that are administered on a less frequent basis (e.g., “maintenance doses”).
  • each secondary and/or tertiary dose is administered 1 to 26 (e.g., 1, 11 ⁇ 2, 2, 21 ⁇ 2, 3, 31 ⁇ 2, 4, 41 ⁇ 2, 5, 51 ⁇ 2, 6, 61 ⁇ 2, 7, 71 ⁇ 2, 8, 81 ⁇ 2, 9, 91 ⁇ 2, 10, 101 ⁇ 2, 11, 111 ⁇ 2, 12, 121 ⁇ 2, 13, 131 ⁇ 2, 14, 141 ⁇ 2, 15, 151 ⁇ 2, 16, 161 ⁇ 2, 17, 171 ⁇ 2, 18, 181 ⁇ 2, 19, 191 ⁇ 2, 20, 201 ⁇ 2, 21, 211 ⁇ 2, 22, 221 ⁇ 2, 23, 231 ⁇ 2, 24, 241 ⁇ 2, 25, 251 ⁇ 2, 26, 261 ⁇ 2, or more) weeks after the immediately preceding dose.
  • 1 to 26 e.g., 1, 11 ⁇ 2, 2, 21 ⁇ 2, 3, 31 ⁇ 2, 4, 41 ⁇ 2, 5, 51 ⁇ 2, 6, 61 ⁇ 2, 7, 71 ⁇ 2, 8, 81 ⁇ 2, 9, 91 ⁇ 2, 10, 101 ⁇ 2, 11, 111 ⁇ 2, 12, 121 ⁇ 2, 13, 131 ⁇ 2, 14, 141 ⁇ 2, 15, 151 ⁇ 2, 16, 161 ⁇ 2, 17, 171 ⁇ 2, 18, 181 ⁇ 2, 19, 19
  • the immediately preceding dose means, in a sequence of multiple administrations, the dose of antigen- binding molecule and/or AAV which is administered to a patient prior to the administration of the very next dose in the sequence with no intervening doses.
  • the methods according to this aspect of the disclosure may comprise administering to a patient any number of secondary and/or tertiary doses of an antigen-binding molecule and/or AAV described herein. For example, in certain embodiments, only a single secondary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, ⁇ ⁇ Attorney Docket No.250298.000954 8, or more) secondary doses are administered to the patient.
  • each secondary dose may be administered at the same frequency as the other secondary doses. For example, each secondary dose may be administered to the patient 1 to 2 weeks after the immediately preceding dose.
  • each tertiary dose may be administered at the same frequency as the other tertiary doses. For example, each tertiary dose may be administered to the patient 2 to 4 weeks after the immediately preceding dose.
  • the frequency at which the secondary and/or tertiary doses are administered to a patient can vary over the course of the treatment regimen.
  • the frequency of administration may also be adjusted during the course of treatment by a physician depending on the needs of the individual patient following clinical examination.
  • the AAV particle when an antigen-binding molecule and an AAV particle described herein are sequentially administered, the AAV particle may be administered as a first component of the dosing regimen and the antigen-binding molecule may be administered as a second component of the dosing regimen (i.e., the AAV particle may be administered before the antigen-binding molecule).
  • the AAV particle may be administered as a second component of the dosing regimen and the antigen-binding molecule may be administered as a first component of a dosing regimen (i.e., the AAV particle may be administered after the antigen-binding molecule).
  • an AAV particle and/or antigen-binding molecule may be sequentially administered, in either of the above-described orders, with variable time intervals between administration.
  • the time interval between administration of the AAV particle and the antigen binding molecule may be ⁇ at least about 30 seconds, at least about 35 seconds, at least about 40 seconds, at least about 45 seconds, at least about 50 seconds, at least about 55 seconds, at least about 1 minute, at least about 2 minutes, at least about 5 minutes, at least about 10 minutes, at least about 20 minutes, at least about 30 minutes, at least about 40 minutes, at least about 50 minutes, at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, at least about 5 hours, at least about 6 hours, at least about 7 hours, at least about 8 hours, at least about 9 hours, at least about 10 hours, at least about 10 to 12 hours, at least about 12 to 14 hours, at least about 14 to 16 hours, at least about 16 to 18 hours, at least about 18 to 20 hours, at least about 20 to 22 hours, at least about 22 to 24 hours, at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 2
  • the present disclosure also provides a vessel (e.g., a vial or chromatography column) or injection device (e.g., syringe, pre-filled syringe, or autoinjector) comprising ⁇ bispecific antigen-binding molecule (e.g., pharmaceutical formulation thereof) set forth herein.
  • the vessel or injection device may be packaged into a kit.
  • An injection device is a device that introduces a substance into the body of a subject (e.g., a human) via a parenteral route, e.g., intraocular, intravitreal, intramuscular, subcutaneous, or intravenous.
  • an injection device may be a syringe (e.g., pre- filled with the pharmaceutical formulation, such as an auto-injector) which, for example, includes a cylinder or barrel for holding fluid to be injected (e.g., comprising the antibody or fragment or a pharmaceutical formulation thereof), a needle for piecing skin, blood vessels, or other tissue for injection of the fluid; and a plunger for pushing the fluid out of the cylinder and through the needle bore and into the body of the subject.
  • a pharmaceutical composition provided herein can be delivered subcutaneously or intravenously with a standard needle and syringe.
  • a pen delivery device readily has applications in delivering a pharmaceutical composition of the present disclosure.
  • Such a pen delivery device can be reusable or disposable.
  • a reusable pen delivery device generally utilizes a replaceable cartridge that contains a pharmaceutical composition. Once all of the pharmaceutical composition within the cartridge has been administered and the cartridge is empty, the empty cartridge can readily be discarded and replaced with a new cartridge that contains the pharmaceutical composition. The pen delivery device can then be reused.
  • a disposable pen delivery device there is no replaceable cartridge. Rather, the disposable pen delivery device comes prefilled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.
  • Numerous reusable pen and autoinjector delivery devices have applications in the subcutaneous delivery of a pharmaceutical composition of the present disclosure.
  • Examples include, but are not limited to AUTOPENTM (Owen Mumford, Inc., Woodstock, UK), DISETRONICTM pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75/25TM pen, HUMALOGTM pen, HUMALIN 70/30TM pen (Eli Lilly and Co., Indianapolis, Ind.), ⁇ ⁇ Attorney Docket No.250298.000954 NOVOPENTM I, II, and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIORTM (Novo Nordisk, Copenhagen, Denmark), BDTM pen (Becton Dickinson, Franklin Lakes, N.J.), OPTIPENTM, OPTIPEN PROTM, OPTIPEN STARLETTM, and OPTICLIKTM (sanofi-aventis, Frankfurt, Germany), to name only a few.
  • Examples of disposable pen delivery devices having applications in subcutaneous delivery of a pharmaceutical composition of the present disclosure include, but are not limited to the SOLOSTARTM pen (sanofi-aventis), the FLEXPENTM (Novo Nordisk), and the KWIKPENTM (Eli Lilly), the SURECLICKTM Autoinjector (Amgen, Thousand Oaks, Calif.), the PENLETTM (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, L.P.), and the HUMIRATM Pen (Abbott Labs, Abbott Park Ill.), to name only a few.
  • a bispecific antigen-binding molecule of the present disclosure comprising introducing e.g., injecting, the molecule into the body of the subject, e.g., with an injection device.
  • Expression Methods Provided herein are recombinant methods for making an antigen-binding molecule of the present disclosure, or an immunoglobulin chain thereof, comprising (i) introducing, into a host cell, one or more polynucleotides encoding light and/or heavy immunoglobulin chains of such an antigen-binding molecule, for example, wherein the polynucleotide is in a vector; and/or integrates into the host cell chromosome and/or is operably linked to a promoter; (ii) culturing the host cell (e.g., mammalian, fungal, Chinese hamster ovary (CHO), Pichia or Pichia pastoris) under conditions favorable to expression of the polynucle
  • a method for making an antigen-binding molecule includes a method of purifying the molecule, e.g., by column chromatography, precipitation, and/or filtration. The product of such a method also forms part of the present disclosure along with a pharmaceutical composition thereof.
  • Host cells comprising an antigen-binding molecule of the present disclosure and/or a polynucleotide encoding immunoglobulin chains of such a molecule (e.g., in a vector) are also part of the present disclosure.
  • Host cells include, for example, mammalian cells such as Chinese hamster ovary (CHO) cells and fungal cells such as Pichia cells (e.g., P. pastoris).
  • mammalian cells such as Chinese hamster ovary (CHO) cells and fungal cells such as Pichia cells (e.g., P. pastoris).
  • AAV adeno-associated virus
  • said antibody, or said antigen-binding fragment thereof comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10;
  • HCVR heavy chain variable region
  • LCVR light chain variable region
  • said antibody, or said antigen-binding fragment thereof comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
  • said antibody, or said antigen-binding fragment thereof comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 ⁇ or 1159, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 ⁇ or 1157; ⁇ ⁇
  • an antibody, or an antigen-binding fragment thereof wherein said antibody, or said antigen-binding fragment thereof, binds to a capsid of an adeno-associated virus (AAV) particle, wherein said antibody, or said antigen- binding fragment thereof, binds to one or more residues in one or more viral protein 3 (VP3) polypeptides in said capsid, wherein said one or more residues are selected from the residues listed in Tables 18-26.
  • AAV adeno-associated virus
  • said antibody, or said antigen-binding fragment thereof binds to a capsid of an AAV2 particle comprising amino acid substitutions R585A and R588A, and wherein the said one or more residues are selected from the following residues in said VP3 polypeptides (according to VP1 numbering): a) S264, A266, N268, H271, N382, S384, Q385, G512, R487, D494, K527, D529, K532, L538, D533, E555, N705, K706, V708, T450, Q457, and R459; b) S264, G265, A266, S267, N268, H271, N382, G383, S384, R487, K527, D528, E530, E574, Q575, K706, and S707; c) N254, L256, K258, I260, S261, Q263, Y272, E322, V32
  • said antibody, or said antigen-binding fragment thereof binds to a capsid of an AAV8 particle, and wherein the said one or more residues are selected from the following residues in said VP3 polypeptides (according to VP1 numbering): a) N255, L257, K259, I261, S262, N263, S266, A269, Y275, E325, V326, T327, Q328, A658, N670, S671, F672, I673, T674, Q548, N549, A550, A551, D559, E720, and T663; and/or b) N255, L257, S266, E325, V326, T327, Q328, N670, S671, F672, I673, T674, N549, ⁇ ⁇ Attorney Docket No.250298.000954 T719, and E720.
  • said antibody, or said antigen-binding fragment thereof binds to a capsid of an AAV9 particle, and wherein the said one or more residues are selected from the following residues in said VP3 polypeptides (according to VP1 numbering): a) N254, I260, N262, Y274, E324, V325, T326, D327, N328, R550, D665, N668, S669, F670, I671, T672, Q546, G547, G549, E718, and A661; and/or b) N254, L256, I260, N262, E324, V325, T326, D551, N668, S669, F670, T672, G330, and E718.
  • said antibody, or said antigen-binding fragment thereof binds to a capsid of an AAV1 particle comprising amino acid substitution N500E. In some embodiments, said antibody, or said antigen-binding fragment thereof, binds to a capsid of an AAV2 particle comprising amino acid substitution(s) R585A and/or R588A. In some embodiments, said antibody, or said antigen-binding fragment thereof, binds to a capsid of an AAV5 particle comprising amino acid substitution T571S.
  • said antibody, or said antigen-binding fragment thereof binds to a capsid of an AAV6 particle comprising amino acid substitution(s) N500E, K531A, and/or K531E. In some embodiments, said antibody, or said antigen-binding fragment thereof, binds to a capsid of an AAV9 particle comprising amino acid substitution(s) N272A and/or W503A.
  • said antibody, or said antigen-binding fragment thereof comprises: 1) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; 2) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 52 ⁇ or 1159, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 ⁇ or 1157; 3) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid
  • said antibody, or said antigen-binding fragment thereof comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 38; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO
  • said antibody, or said antigen-binding fragment thereof comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 ⁇ or 1159, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 ⁇ or 1157; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 71, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; or 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 81, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • said antibody, or said antigen-binding fragment thereof is a humanized antibody or antigen binding fragment thereof, human antibody or antigen binding fragment thereof, ⁇ ⁇ Attorney Docket No.250298.000954 murine antibody or antigen binding fragment thereof, chimeric antibody or antigen binding fragment thereof, Fab fragment, F(ab')2 fragment, F(ab)'3 fragments, single-chain fragment variable (scFv), bis-scFv, (scFv)2, diabody, minibody, nanobody, triabody, tetrabody, disulfide stabilized Fv protein (dsFv), single-domain antibody (sdAb), Ig NAR, single heavy chain antibody, bispecific antibody, trispecific antibody, or chemically modified derivatives thereof.
  • said antibody, or said antigen-binding fragment thereof comprises an IgG1, IgG2, IgG3, or IgG4 constant region, or a variant thereof.
  • said antibody, or said antigen-binding fragment thereof comprises an IgG1 constant region, or a variant thereof.
  • the IgG1 constant region comprises the amino acid sequence of SEQ ID NO: 259 ⁇ or 1164.
  • said antibody, or said antigen-binding fragment thereof comprises an IgG4 constant region, or a variant thereof.
  • the IgG4 variant comprises a hinge domain modified to reduce binding to Fc ⁇ receptors.
  • the IgG4 constant region comprises the amino acid sequence of SEQ ID NO: 261.
  • said antigen-binding fragment is a Fab fragment.
  • said antigen-binding fragment is an scFv fragment.
  • said antibody, or said antigen-binding fragment thereof binds to AAV2 with a dissociation constant (KD) of less than about 1.5 x 10 -6 M, as measured in a surface plasmon resonance assay.
  • said antibody, or said antigen-binding fragment thereof binds to AAV2 with a dissociation constant (KD) of less than about 20 nM, as measured in a surface plasmon resonance assay.
  • said capsid comprises one or more wild-type AAV capsid polypeptides.
  • said capsid comprises one or more non-wild-type AAV capsid polypeptides.
  • rein binding of said antibody or antigen-binding fragment thereof to said AAV particle capsid does not neutralize said AAV particle.
  • an antibody or an antigen-binding ⁇ ⁇ Attorney Docket No.250298.000954 fragment thereof, that competes for binding to a capsid of an adeno-associated virus (AAV) particle with any of the above-described antibodies or antigen-binding fragments.
  • AAV adeno-associated virus
  • a pharmaceutical composition comprising any of the above-described antibodies or antigen-binding fragments and a pharmaceutically acceptable carrier or excipient.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: i) a first antigen-binding domain that binds to a capsid of an adeno-associated virus (AAV) particle; and ii) a second antigen-binding arm that binds to a molecule on a cell surface
  • said first antigen-binding domain comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10;
  • a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO:
  • the first antigen-binding domain that binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO:
  • the first antigen-binding domain that binds to a capsid of an AAV particle comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; ⁇ ⁇ Attorney Docket No.250298.000954 b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 ⁇ or 1159, and/or
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: i) a first antigen-binding domain that binds to a capsid of an adeno-associated virus (AAV) particle, and ii) a second antigen-binding domain that binds to a molecule on a cell surface, wherein said first antigen-binding domain binds to one or more residues in one or more viral protein 3 (VP3) polypeptides in said capsid, wherein said one or more residues are selected from the residues listed in Tables 18-26.
  • AAV adeno-associated virus
  • said first antigen-binding domain binds to a capsid of an AAV2 particle comprising amino acid substitutions R585A and R588A, and wherein the said one or more residues are selected from the following residues in said VP3 polypeptides (according to VP1 numbering): a) S264, A266, N268, H271, N382, S384, Q385, G512, R487, D494, K527, K529, K532, L538, D533, E555, N705, K706, V708, T450, Q457, and R459; b) S264, G265, A266, S267, N268, H271, N382, G383, S384, R487, K527, D528, E530, Q574, Q575, K706, and S707; c) N254, L256, K258, I260, S261, Q263, Y272, E322, V323, T324, Q32
  • said first antigen-binding domain binds to a capsid of an AAV8 particle, and wherein the said one or more residues are selected from the following residues in said VP3 polypeptides (according to VP1 numbering): e) N255, L257, K259, I261, S262, N263, S266, A269, Y275, E325, V326, T327, Q328, A658, N670, S671, F672, I673, T674, Q548, N549, A550, A551, D559, E720, and T663; and/or f) N255, L257, S266, E325, V326, T327, Q328, N670, S671, F672, I673, T674, N549, T719, and E720.
  • said first antigen-binding domain binds to a capsid of an AAV9 particle, and wherein the said one or more residues are selected from the following residues in said VP3 polypeptides (according to VP1 numbering): g) N254, I260, N262, Y274, E324, V325, T326, D327, N328, R550, D665, N668, S669, F670, I671, T672, Q546, G547, G549, E718, and A661; and/or h) N254, L256, I260, N262, E324, V325, T326, D551, N668, S669, F670, T672, G330, and E718.
  • said first antigen-binding domain that binds to a capsid of an AAV particle comprises: 1) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; 2) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 52 ⁇ or 1159, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 ⁇ or 1157; 3) a heavy chain variable region (HCVR) that comprises a HCDR1,
  • said first antigen-binding domain that binds to a capsid of an AAV particle comprises : 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 38; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 54, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 56, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 58; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and a
  • said first antigen-binding domain that binds to a capsid of an AAV particle comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 or 1159, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 ⁇ or 1157; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 71, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; or 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 81, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary ⁇ ⁇ Attorney Docket No.250298.000954 subunit gamma 1 (CACNG1).
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary ⁇ ⁇ Attorney Docket No.250298.000954 subunit gamma 1
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary ⁇ ⁇ Attorney Docket No.250298.000954 subunit gamma 1
  • the second antigen-binding domain that binds to ASGR1 comprises: a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • said second antigen-binding domain that binds to ASGR1 comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 222, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 226; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 232, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 234, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 236; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and
  • said second antigen-binding domain that binds to ASGR1 comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the molecule on the cell surface is TfR.
  • the second antigen-binding domain that binds to TfR comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino ⁇ ⁇ Attorney Docket No.250298.000954 acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or
  • said second antigen-binding domain that binds to TfR comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 288, a LCDR2 comprising the amino acid sequence of SEQ ID NO:
  • the second antigen-binding domain that binds to TfR comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 314, and/or a LCVR comprising the amino acid sequence of SEQ ID NO:
  • the molecule on the cell surface is CACNG1.
  • the antigen-binding domain that binds to CACNG1 comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HC
  • the second antigen-binding domain that binds to CACNG1 comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 645, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 6
  • the second antigen-binding domain that binds to CACNG1 comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 703; 6)
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); and c) a third polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form Fab2, wherein Fab2 comprises a second antigen-binding domain (“ABD2”); wherein ABD1 or ABD2 binds to a capsid of an adeno-associated virus (AAV) particle, and the other of ABD1 or ABD2 binds to
  • ABD1 binds to a capsid of an adeno-associated virus (AAV) particle and ABD2 binds to a molecule on the cell surface.
  • ABD2 binds to a capsid of an adeno-associated virus (AAV) particle and ABD1 binds to a molecule on the cell surface.
  • the first heavy chain region is linked to the second heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first scFv ⁇ ⁇ Attorney Docket No.250298.000954 comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) an Fc domain; b) a second polypeptide chain, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain; and c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); wherein ABD1 or ABD2 binds to a capsid of an adeno-associated virus (AAV) particle, and
  • AAV adeno-associated virus
  • ABD1 binds to a capsid of an adeno-associated virus (AAV) particle and ABD2 binds to a molecule on the cell surface.
  • ABD2 binds to a capsid of an adeno-associated virus (AAV) particle and ABD1 binds to a molecule on the cell surface.
  • the first scFv and/or the first heavy chain region is linked to the Fc domain via a linker.
  • the ABD1 or ABD2 which binds to a capsid of an AAV particle comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and
  • the ABD1 or ABD2 which binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 8.
  • the ABD1 or ABD2 which binds to a capsid of an AAV particle comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 or 1159, and/or a
  • the molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary subunit gamma 1 (CACNG1).
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • the other of ABD1 or ABD2 which binds to ASGR1 comprises: a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the other of ABD1 or ABD2 which binds to ASGR1 comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 222, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 226; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 232, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 234, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 236; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO:
  • the other of ABD1 or ABD2 which binds to ASGR1 comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the molecule on the cell surface is TfR.
  • the other of ABD1 or ABD2 which binds to TfR comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or
  • the other of ABD1 or ABD2 which binds to TfR comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 288, a LCDR2 comprising the amino acid sequence of SEQ ID NO:
  • the other of ABD1 or ABD2 which binds to TfR comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; ⁇ ⁇ Attorney Docket No.250298.000954 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 314, and
  • the molecule on the cell surface is CACNG1.
  • the other of ABD1 or ABD2 which binds to CACNG1 comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid ⁇ ⁇ Attorney Docket No.250298.000954 sequence of SEQ ID NO: 643; 3) a HCVR that comprises the HCDR1,
  • the other of ABD1 or ABD2 which binds to CACNG1 comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 645, a LCDR2 comprising
  • the other of ABD1 or ABD2 which binds to CACNG1 comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR ⁇ ⁇ Attorney Docket No.250298.000954 comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/or a LCVR
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) a first heavy chain region of a first Fab (“Fab1”), said first heavy chain region operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d) a fourth polypeptide chain comprising a second light chain that pairs
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the scFv is linked to the first heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) an scFv comprising a first antigen-binding domain (“ABD1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d) a fourth polypeptide chain comprising a second light chain that pairs with
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the scFv is linked to the Fc domain via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”), said second heavy chain region of Fab2 operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); d) a fourth polypeptide chain comprising a second light chain that pairs with the second
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the first heavy chain region is linked to the second heavy chain region via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an scFv comprising a first antigen-binding domain (“ABD1”), said scFv operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain ⁇ ⁇ Attorney Docket No.250298.000954 (“ABD2”); and d
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 binds to a molecule on a cell surface.
  • the scFv is linked to the first heavy chain region via a linker.
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a second heavy chain region of a second Fab (“Fab2”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a second antigen-binding domain (“ABD2”); and d) a fourth polypeptide chain comprising a second light chain that pairs with
  • ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ⁇ ⁇ Attorney Docket No.250298.000954 ABD3 binds to a molecule on a cell surface.
  • the second heavy chain region is linked to the Fc domain via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”), said second heavy chain region operably linked to (iii) a third heavy chain region of a third Fab (“Fab3”); b) a second polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form Fab1, wherein Fab1 comprises a first antigen-binding domain (“ABD1”); and c) a third polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form Fab2, wherein Fab2 comprises a second antigen
  • two of ABD1, ABD2, and ABD3 bind to a capsid of an AAV particle, and one of ABD1, ABD2, and ABD3 binds to a molecule on a cell surface.
  • ABD2 and ABD3 bind to a capsid of an AAV particle, and ABD1 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 binds to a molecule on a cell surface.
  • the first heavy chain region is operably linked to the second heavy chain region and/or the second heavy chain region is operably linked to the third heavy chain region via a linker.
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and ⁇ ⁇ Attorney Docket No.250298.000954 HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence
  • the at least one of ABD1, ABD2, and ABD3 which binds to a capsid of an AAV particle comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 ⁇ or 1159, and/or a LCVR comprising the amino acid
  • said molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary subunit gamma 1 (CACNG1). [001071] In some embodiments, said molecule on the cell surface is ASGR1.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises: a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 222, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 226; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 232, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 234, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 236; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ASGR1 comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the molecule on the cell surface is TfR.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to ⁇ ⁇ Attorney Docket No.250298.000954 TfR comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 288, a LCDR2 comprising the
  • the other(s) of ABD1, ABD2, and ABD3 which binds to TfR comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 314, and/or a LCVR comprising the
  • the molecule on the cell surface is CACNG1.
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO:
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 645,
  • the other(s) of ABD1, ABD2, and ABD3 which binds to CACNG1 comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/or a LCVR comprising the amino acid sequence of SEQ ID NO:
  • a multispecific antibody or a multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first scFv comprising a first antigen-binding domain (“ABD1”) operably linked to (ii) a first heavy chain region of a first Fab (“Fab1”), said first heavy chain region of Fab1 operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second scFv comprising a second antigen-binding domain (“ABD2”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”), said second heavy chain region of Fab2 operably linked to (iii) an Fc domain; c) a third polypeptide chain comprising a first light chain that pairs with the first
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD3 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD2 binds to a molecule on a cell surface.
  • the first scFv is linked to the first heavy chain region and/or the second scFv is linked to the second heavy chain region via a linker.
  • a multispecific antibody, or a multispecific antigen-binding fragment thereof comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a first scFv comprising a first antigen-binding domain (“ABD1”); b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab (“Fab2”) operably linked to (ii) an Fc domain, said Fc domain operably linked to (iii) a second scFv
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD2 bind to a capsid of an AAV particle, and ABD3 and ABD4 binds to a molecule on a cell surface.
  • ABD3 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD2 binds to a molecule on a cell surface.
  • the first scFv and/or second scFv is linked to the Fc domain via a linker.
  • a multispecific antibody or a ⁇ ⁇ Attorney Docket No.250298.000954 multispecific antigen-binding fragment thereof, comprising: a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) a first heavy chain region of a first Fab (“Fab1”) operably linked to (ii) a second heavy chain region of a second Fab (“Fab2”) operably linked to (iii) an Fc domain; b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a third heavy chain region of a third Fab (“Fab3”) operable liked to (ii) a fourth heavy chain region of a fourth Fab (“Fab4”) operably linked to (iii) an Fc domain; c)
  • two of ABD1, ABD2, ABD3, and ABD4 bind to a capsid of an AAV particle, and two of ABD1, ABD2, ABD3, and ABD4 binds to a molecule on a cell surface.
  • ABD1 and ABD3 bind to a capsid of an AAV particle, and ABD2 and ABD4 binds to a molecule on a cell surface.
  • ABD2 and ABD4 bind to a capsid of an AAV particle, and ABD1 and ABD3 binds to a molecule on a cell surface.
  • the first heavy chain region is linked to the second heavy chain region and/or the third heavy chain region is linked to fourth heavy chain region via a linker.
  • the at least one of ABD1, ABD2, ABD3, and ABD4 which binds to a capsid of an AAV particle comprises: a) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a light chain variable region (LCVR) that comprises a LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10; ⁇ ⁇ Attorney Docket No.250298.000954 b) a heavy chain variable region (HCVR) that comprises a HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or
  • the at least one of ABD1, ABD2, ABD3, and ABD4 which binds to a capsid of an AAV particle comprises: a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID
  • the at least one of ABD1, ABD2, ABD3, and ABD4 which binds to a capsid of an AAV particle comprises: a) a HCVR comprising the amino acid sequence of SEQ ID NO: 2, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; b) a HCVR comprising the amino acid sequence of SEQ ID NO: 22, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; c) a HCVR comprising the amino acid sequence of SEQ ID NO: 32, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; d) a HCVR comprising the amino acid sequence of SEQ ID NO: 42, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10; e) a HCVR comprising the amino acid sequence of SEQ ID NO: 52 or 1159, and/or a LCVR comprising the amino amino acid sequence of SEQ ID NO
  • said molecule on the cell surface is asialoglycoprotein receptor 1 (ASGR1), transferrin receptor (TfR), or calcium voltage-gated channel auxiliary subunit gamma 1 (CACNG1).
  • ASGR1 asialoglycoprotein receptor 1
  • TfR transferrin receptor
  • CACNG1 calcium voltage-gated channel auxiliary subunit gamma 1
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to ASGR1 comprises: a) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to ASGR1 comprises a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 222, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 224, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 226; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 232, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 234, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 236; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, a LCDR
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to ASGR1 comprises a) a HCVR comprising the amino acid sequence of SEQ ID NO: 220 or 1239, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10 or 1157; or b) a HCVR comprising the amino acid sequence of SEQ ID NO: 230, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 10.
  • said molecule on the cell surface is TfR.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to TfR comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to TfR comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 272, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 273, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 274; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 277, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 279; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 283, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 284, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 285; and/or a LCDR1 comprising the amino acid sequence of ⁇ ⁇ Attorney Docket No.250298
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to TfR comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 270 or 271, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 276; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 281 or 282, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 287; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 292 or 293, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 298; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 303, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 308 or 309; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 314, and/or a LCVR comprising the amino acid sequence of SEQ
  • said molecule on the cell surface is CACNG1.
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to CACNG1 comprises: 1) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR that comprises the HCDR1, HCDR2, and HCDR3 contained within an HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR that comprises the LCDR1, LCDR2, and LCDR3 contained within a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR that comprises the HCDR1, HCDR2, and HCDR3
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to CACNG1 comprises: 1) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 617, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 619, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 621; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 627, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 629; 2) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 637, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 641; and/or a LCDR1 comprising the amino acid sequence of SEQ ID NO: 6
  • the other(s) of ABD1, ABD2, ABD3, and ABD4 which binds to CACNG1 comprises: 1) a HCVR comprising the amino acid sequence of SEQ ID NO: 615, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 623; 2) a HCVR comprising the amino acid sequence of SEQ ID NO: 635, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 643; 3) a HCVR comprising the amino acid sequence of SEQ ID NO: 655, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 663; 4) a HCVR comprising the amino acid sequence of SEQ ID NO: 675, and/or a LCVR comprising the amino acid sequence of SEQ ID NO: 683; 5) a HCVR comprising the amino acid sequence of SEQ ID NO: 695, and/or a LCVR comprising the amino acid sequence of S
  • the linker is: (a) at least 5 amino acids, at least 6 amino acids, or at least 7 amino acids in length; and optionally (b) up to 30 amino acids, up to 40 amino acids, up to 50 amino acids, or up to 60 amino acids in length.
  • the linker is: (a) 5 amino acids to 50 amino acids in length; (b) 5 amino acids to 45 amino acids in length; (c) 5 amino acids to 40 amino acids in length; (d) 5 amino acids to 35 amino acids in length; (e) 5 amino acids to 30 amino acids in length; amino acids to 25 amino acids in length; or amino acids to 20 amino acids in length.
  • the linker is: amino acids to 50 amino acids in length; amino acids to 45 amino acids in length; ⁇ ⁇ Attorney Docket No.250298.000954 (c) 6 amino acids to 40 amino acids in length; (d) 6 amino acids to 35 amino acids in length; (e) 6 amino acids to 30 amino acids in length; (f) 6 amino acids to 25 amino acids in length; or (g) 6 amino acids to 20 amino acids in length. [001117] In some embodiments, the linker is: (a) 7 amino acids to 40 amino acids in length; (b) 7 amino acids to 35 amino acids in length; amino acids to 30 amino acids in length; amino acids to 25 amino acids in length; amino acids to 20 amino acids in length.
  • the linker is 5 amino acids to 25 amino acids in length. In some embodiments, the linker is 10 amino acids to 60 amino acids in length. In some embodiments, the linker is 20 amino acids to 50 amino acids in length. In some embodiments, the linker is 25 to 35 amino acids in length. In some embodiments, the linker is or comprises a multimer of GnS (SEQ ID NO: (SEQ ID NO: 240), wherein n is an integer from 1 to 10.
  • the linker is or comprises a multimer of G 4 S (SEQ ID NO:
  • the linker is or comprises two consecutive glycines (2Gly), three consecutive glycines (3Gly), four consecutive glycines (4Gly (SEQ ID NO: 243)), five consecutive glycines (5Gly (SEQ ID NO: 244)), six consecutive glycines (6Gly (SEQ ID NO: 245)), seven consecutive glycines (7Gly (SEQ ID NO: 246)), eight consecutive glycines (8Gly (SEQ ID NO: 247)), or nine consecutive glycines (9Gly (SEQ ID NO: 248)).
  • one or more light chains are universal light chains.
  • a light chain constant region (CL) and a first heavy chain constant region (CH1) of one or more Fabs are in a CrossmAb arrangement.
  • the multispecific antibodies or multispecific antigen- binding fragments comprise an Fc heterodimer.
  • the Fc domains in the Fc heterodimer comprise knob-in- hole mutations as compared to a wild-type Fc domain.
  • one Fc domain in the Fc heterodimer comprises amino acid substitutions S354C and T366W (according to EU numbering) as compared to a wild- type Fc domain
  • the other Fc domain in the Fc heterodimer comprises amino acid substitutions Y349C, T366S, L368A, and Y407V (according to EU numbering) as compared ⁇ ⁇ Attorney Docket No.250298.000954 to a wild-type Fc domain.
  • at least one Fc domain in the Fc heterodimer comprises a star mutation as compared to a wild-type Fc domain.
  • one Fc domain in the Fc heterodimer comprises amino acid mutations H435R and/or Y436F (according to EU numbering) as compared to a wild-type Fc domain.
  • said capsid comprises one or more wild-type AAV capsid polypeptides.
  • said capsid comprises one or more non-wild-type AAV capsid polypeptides.
  • the multispecific antibodies or multispecific antigen-binding fragments can be a bispecific antibody or bispecific antigen-binding fragment thereof.
  • a pharmaceutical composition comprising a multispecific antibody or multispecific antigen-binding fragment described herein, and a pharmaceutically acceptable carrier or excipient.
  • a molecular complex comprising an AAV particle bound to one or more antibodies and/or antigen-binding fragments described herein.
  • said AAV particle comprises one or more mutations in one or more AAV capsid proteins inhibiting the natural tropism of said AAV particle.
  • a pharmaceutical composition comprising a molecular complex described herein and a pharmaceutically acceptable carrier or excipient.
  • a method of preparing the molecular complex described herein comprising incubating said AAV particle in the presence of said one or more antibodies and/or antigen-binding fragments under conditions allowing specific binding of said one or more antibodies and/or antigen-binding fragments to said AAV particle capsid.
  • a molecular complex comprising an AAV particle bound to one or more multispecific antibodies and/or multispecific antigen- binding fragments described herein.
  • said AAV particle comprises one or more mutations in one or more AAV capsid proteins inhibiting the natural tropism of said AAV particle.
  • a pharmaceutical composition comprising a molecular complex described herein and a pharmaceutically acceptable carrier or excipient.
  • a pharmaceutically acceptable carrier or excipient ⁇ ⁇ Attorney Docket No.250298.000954
  • a method of preparing a molecular complex described herein comprising incubating said AAV particle in the presence of said one or more multispecific antibodies and/or multispecific antigen-binding fragments under conditions allowing specific binding of said one or more multispecific antibodies and/or multispecific antigen-binding fragments to said AAV particle capsid.
  • a method for targeting an AAV particle to a cell expressing a molecule on the cell surface comprising contacting the cell with a molecular complex described herein or a pharmaceutical composition described herein, wherein said molecular complex comprises one or more multispecific antibodies and/or multispecific antigen-binding fragments which bind to said molecule on the cell surface.
  • a method for delivering a polynucleotide to a cell expressing a molecule on the cell surface comprising contacting the cell with a molecular complex described herein or a pharmaceutical composition described herein, wherein said molecular complex comprises the AAV particle comprising said polynucleotide and bound to one or more multispecific antibodies and/or multispecific antigen- binding fragments which bind to said molecule on the cell surface.
  • said cell is in a subject and said molecular complex is administered to the subject.
  • said AAV particle does not target said cell in the absence of said one or more multispecific antibodies and/or multispecific antigen-binding fragments.
  • the AAV capsid structure comprises three viral proteins, VP1, VP2, and VP3, that are encoded from a single open reading frame.
  • An icosahedral capsid is formed from the ⁇ ⁇ Attorney Docket No.250298.000954 assembly of 60 monomers of VP1, VP2, and VP3 in relative amounts of 1:1:10.
  • the AAV capsid contains residues critical for mediating cell and receptor binding.
  • mAbs Monoclonal antibodies to AAV have been generated and the ability of these mAbs to bind to AAV capsids of various serotypes was explored by enzyme-linked immunosorbent assay (ELISA).
  • ELISA enzyme-linked immunosorbent assay
  • Anti-AAV Antibody Identifiers [001151] Anti-AAV mAbs of the present Example (HDQQ06E09-0017, HDQQ06F06-0018, HDPV07D04-0065, 4059311_ULC1-39_38741_MS, 3842258_ULC1-3, _79284_MS, 4059310_ULC1-39_48703_MS, 4059311_ULC1-39_31283_MS, 4059311_ULC1- 39_36126_MS, 4059311_ULC1-39_31000_MS, 4059310_ULC1-39_51231_MS) were assessed for binding to multiple AAV serotypes (AAV2 HBM [heparin binding mutant], AAV3b, AAV5, AAV6, AAV8, AAV9, AAV Sealion, AAV rh32.33).
  • the viral particles were first diluted in PBS and used to coat black 96-well Maxisorp plates (2x10 8 VGs/well) overnight at 4 °C. The plates were then emptied, pat-dried, washed 4x with KPL 1x Wash Buffer, and incubated with 3%-bovine serum albumin (BSA) blocking buffer overnight at 4 °C.
  • BSA 3%-bovine serum albumin
  • the bivalent antibodies were diluted 1:8 in assay diluent buffer (ADB) from a starting concentration of 5 ⁇ g/ml. The blocked plates were then emptied, pat-dried, washed 4x, and incubated with the bivalent antibodies for 1 hour at room temperature (RT).
  • REGN13076 was able to bind to all serotypes.
  • REGN13072 bound strongly to all serotypes except for AAV rh32.33.
  • Two hybridoma-derived antibodies (REGN13070, REGN13071) and two NGS/MS-derived antibodies (REGN13073, REGN13074) showed strong binding to AAV2 HBM and AAV3b, but weak binding to the remaining serotypes.
  • REGN13075 and REGN13220 bound to multiple serotypes with varying strength.
  • REGN13284 bound most strongly to AAV8.
  • REGN13221 showed weak affinity to all serotypes at low concentrations. However, binding for REGN13221 was observed with increased amount of antibody.
  • Example 2
  • Binding kinetics of anti-AAV antibodies to surface-coupled AAV-2 [001153] Equilibrium dissociation constants (K D values) for adeno-associated virus serotype- 2 (AAV-2) binding to purified anti-AAV antibodies and the respective Fab domain proteins were determined using a real-time surface plasmon resonance biosensor technology using a Biacore T200 instrument. [001154] Table 13 shows a description of identifiers for anti-AAV antibodies of the present Example. Table 13. Anti-AAV Antibody Identifiers ⁇ ⁇ Attorney Docket No.250298.000954 REGN13885-L1 [001155] Table 14 shows a description of the reagents used in the present Example. Table 14.
  • CM5 Biacore sensor surface was derivatized by amine coupling with adeno- associated virus (AAV-2) around 850 RU. All Biacore binding studies were performed in a buffer composed of 0.01M HEPES, 0.15 M NaCl, pH 7.4 (HBS-N running buffer). Different concentrations of anti-AAV antibodies (ranging from 100 nM to 25 nM in 4-fold serial dilutions) prepared in HBS-N running buffer were injected over the amine coupled AAV-2 at a flow rate of 30 ⁇ L/minute. Antibody association was monitored for 2.5 minutes while dissociation was monitored in HBS-N running buffer for 5 minutes.
  • AAV-2 adeno- associated virus
  • the AAV-2 capture surface was regenerated using two 10 second injections of 10mM Glycine at pH 1.5. All binding kinetics experiments were performed at 25 °C. [001157]
  • the specific SPR-Biacore sensorgrams were obtained by a double referencing procedure. The double referencing was performed by first subtracting the signal of each injection over a reference surface (blank amine coupled surface) from the signal over the experimental surface (amine coupled AAV-2) thereby removing contributions from refractive index changes. In addition, running buffer injections were performed to allow subtraction of the signal changes resulting from the dissociation phase.
  • Antibodies can be used to bridge AAV capsids to target cells to enhance transduction efficiency.
  • the high affinity immunoglobulin gamma Fc Receptor I (FcGR1) binds to the Fc of various IgG isotypes, including IgG1, and can be expressed on cell lines to capture antibodies on cell surface.
  • FcGR1 immunoglobulin gamma Fc Receptor I
  • mAbs monoclonal antibodies
  • Table 17 shows a description of identifiers for anti-AAV antibodies of the present Example. Table 17.
  • Anti-AAV Antibody Identifiers [001162] Anti-AAV mAbs of the present Example (HDQQ06E09-0017, HDQQ06F06-0018, HDPV07D04-0065, 4059311_ULC1-39_38741_MS, 3842258_ULC1-3, _79284_MS, 4059310_ULC1-39_48703_MS, 4059311_ULC1-39_31283_MS, 4059311_ULC1- 39_36126_MS, 4059311_ULC1-39_31000_MS, 4059310_ULC1-39_51231_MS) were assessed for their ability to enhance internalization of multiple AAV serotypes expressing NanoLuc genomes (AAV1, AAV2.Myc, AAV5, AAV8, AAV9, AAV Sealion) into FcGR1- expressing HEK 293T cells.
  • NanoLuc genomes AAV1, AAV2.Myc, AAV5, AAV8, AAV
  • Antibodies were first diluted in 1 x PBS for appropriate viral genome to antibody ratios (1:4, 1:500, 1:2500, 1:10,000, undiluted) and added to 96-well CoStar Assay black with clear bottom assay plates. Afterwards, AAVs were added to plates containing antibodies and incubated for 1 hour at 37 °C and 5% CO2. A suspension of HEK 293T FcGR1+ cells in DMEM (supplemented with 10% fetal bovine serum [FBS], minimum essential medium (MEM) non-essential amino acid (NEAA) solution, Penicillin-Streptomycin [Pen Strep]) was then added to virus and antibody complexes.
  • FBS fetal bovine serum
  • MEM minimum essential medium
  • NEAA non-essential amino acid
  • Penicillin-Streptomycin [Pen Strep] Penicillin-Streptomycin
  • transduced cells were incubated for 72 hours, and transduction was measured using the NanoGlo Luciferase assay system. Luminescence was detected using a plate reader (Tecan Infinite M200 Pro) and analyzed using GraphPad Prism. [001163] As shown in Figures 2A-2J, five of the antibodies (4059311_ULC1-39_38741_MS, 4059311_ULC1-39_31000_MS, 4059311_ULC1-39_36126_MS, HDQQ06E09-0017, HDPV07D04-0065) exhibited enhanced transduction at lower antibody concentrations for the majority of serotypes tested.
  • Cryo-electron Microscopy (Cryo-EM) [001164] Cryo-EM sample preparation and data collection [001165] Purified anti-AAV Fabs were mixed with corresponding AAV samples ( ⁇ 1.5x10 13 to 2.5 x10 14 VG/ml) at approximately 220:1 molar ratio and incubated at 4 °C for 1 hour.3.5 ⁇ L of the mixture was applied onto an UltrGermanoil R1.2/1.3, 300 mesh grid (Quantifoil). Excess liquid was blotted away using filter paper and the grid was plunge frozen in liquid ethane cooled by liquid nitrogen using a Vitrobot Mark IV (ThermoFisher) operated at 4 °C and 100% humidity.
  • Junk particles were removed by multiple rounds of 2D classification, followed by Ab initio reconstruction, Homogeneous refinement, and Heterogenous refinement to identify the best class of particles representing the target complex. These particles were further refined using non-uniform refinement to generate the final cryo-EM density map. The details of data processing for each sample are summarized in Table 18. [001168] Model building and refinement [001169] Manual model building was carried out using Coot version 0.8.9 and real space refinements were done in Phenix version 1.17. Published high-resolution structures of AAV2 (PDB: 6NZ0), AAV8 (PDB: 2QA0), and AAV9 (PDB: 3UX1) were used as initial models for model building.
  • Homology models for Fab fragments used in this Example were made from a previously determined REGN Fab structure. Unambiguous docking of Fab models into their respective densities was aided by clearly interpretable side chain densities corresponding to their distinct complementarity determining region (CDR) sequences. A combination of picked initial models was docked into corresponding cryo-EM density map using Fit-in-map of UCSF Chimera. The models were adjusted manually in Coot, followed by real space ⁇ ⁇ Attorney Docket No.250298.000954 refinement with secondary structure and non-crystallographic symmetry (NCS) restraints in Phenix.
  • NCS non-crystallographic symmetry
  • REGN13878 largely binds to the edge of one VP3 trimer core (trimer core #1) but also tilts towards another adjacent VP3 trimer core (trimer core #2), making extensive contacts with 5 VP3 protomers in the two trimer cores.
  • trimer core #1 the VP3 protomers in trimer core #1 were named as VP3-3a, VP3-3b, and VP3-3c in clockwise direction around its 3-fold axis.
  • the VP3 protomers in the adjacent trimer core #2 were named as VP3-3d, VP3-3e, and VP3-3f in clockwise direction around its 3-fold axis, where VP3-3d shares a 5-fold axis with VP3-3a and VP3-3d is in the counterclockwise direction of VP3-3a around this 5-fold axis.
  • REGN13878 residues contacting VP3 residues of AAV2 HBM are summarized in Table 19. The binding of REGN13878 is primarily mediated by its heavy chain contacting VP3-3a and VP3-3b in trimer core #1 and is further stabilized by engagement of the light chain with VP3- 3d and VP3-3e in trimer core #2.
  • CDRs H1, H2, and H3 of REGN13878 engages VP3-3a by contacting two patches of residues located in variable region I (VR-1) (S264, A266, N268) and VR-III (N382, S384, Q385), as well as two additional residues H271 and G512.
  • CDRs H2 and H3 of REGN13878 also contacts VP3-3b VR-V (R487, D494), VR-VI (K527, D529, K532).
  • a single residue from VR-VI of VP3-3c, L538, is also involved in the interaction with CDRs H2 of REGN13878.
  • CDRs H3, L1, and L2 contacts a patch of residues in VP3-3d VR-IX (N705, K706, V708), with addition interactions mediated by VR-VI (D533, E555) and the framework region (FR) of the light chain.
  • VR-IV residues (T450, Q457, R459) in VP3-3e contacts REGN13878 light chain FR as well.
  • Cryo-EM structure of AAV2 HBM in complex with REGN13883 [001175] A 3.07 ⁇ cryo-EM structure of AAV2 HBM in complex with the anti-AAV antibody Fab REGN13883 was obtained.
  • REGN13883 binds around the icosahedral 3-fold axis. Since the binding site is located at the junction of two adjacent trimer cores which both have three VP3 protomers at two adjacent 3-fold axes, the binding site is also around the icosahedral 2-fold axis. ⁇ ⁇ Attorney Docket No.250298.000954 [001176] REGN13883 binds to the boundary of two adjacent VP3 trimer cores, trimer core #1 and trimer core #2, but the interactions are largely mediated by trimer core #1. The Fab is roughly perpendicular to the virus capsid surface.
  • the VP3 protomers in trimer core #1 and the adjacent trimer core #2 were named following the same convention used in the AAV2 HBM-REGN13878 structure.
  • REGN13883 residues contacting VP3 residues of AAV2 HBM are summarized in Table 20.
  • the binding of REGN13883 is primarily mediated by the heavy chain contacting VP3-3a and VP3-3b in trimer core #1 and is further stabilized by light chain contacts with VP3-3d in trimer core #2.
  • CDRs H1, H2, H3, and L2 of REGN13883 engages VP3-3a by contacting two patches of residues located in variable region I (VR-I) (S264, G265, A266, S267, N268) and VR-III (N382, G383, S384), as well as an additional H271 residue.
  • CDRs H2 and H3 of REGN13883 also contacts VP3-3b VR-VI (K527, D528, E530), VR-V (R487), and two additional residues (E574, Q575).
  • CDR L1 of REGN13883 contacts a patch of residues in VP3-3d VR-IX (K706, S707) to stabilize the binding.
  • REGN13880 residues contacting VP3 residues of AAV2 HBM are summarized in Table 21.
  • the binding of REGN13880 is primarily mediated by heavy chain contacts with VP3-5a and light chain contacts with VP3-5b; the heavy chain makes additional contacts with VP3-5e.
  • REGN13880 has a very long CDR H3 that engages the DE loop (E322, V323, T324, Q325) and HI loop (A655, A667, S668, F669, I670, T671) of VP3-5a.
  • CDRs H1, H2, and H3 of REGN13880 also engage VP3-5a by contacting a patch of residues spanning VR- 1 (N254, L256, K258, I260, S261, Q263, Y272).
  • the binding is further stabilized by CDRs L1, L3, and H3 of REGN13880 contacting VP3-5b VR-VII (Q545, G546, S547, E548, K556) and three additional residues (I332, N717, and V719).
  • Two residues from VP3-5e, T660 and F661, are also involved in the interaction with CDR H1 of REGN13880. Since the epitope of REGN13880 is primarily located at the conserved regions of VP3, the Fab can also bind to other AAV serotypes including AAV8 and AAV9 as described below.
  • REGN13880 residues contacting VP3 residues of AAV8 were summarized in Table 22.
  • the binding of REGN13880 is also primarily mediated by its heavy chain contacting VP3-5a and light chain contacting VP3-5b, with the heavy chain making additional contacts with VP3-5e.
  • the long CDR H3 of REGN13880 dominates the interactions by engaging the DE loop (E325, V326, T327, Q328) and HI loop (A658, N670, S671, F672, I673, T674) of VP3-5a as well, with an additional contact between VP3-5a N670 and CDR L1.
  • CDRs H1, H2, and H3 of REGN13880 also engage VP3-5a by contacting a patch of residues spanning VR-I (N255, L257, K259, I261, S262, N263, A269, S266, Y275). The binding is further stabilized by CDRs L1 and L3 of REGN13880 contacting VP3-5b VR-VII (Q548, N549, A550, A551, D559) and an additional residue E720.
  • One residue from VP3-5e, T663, is also involved in the interaction with CDR H1 of REGN13880.
  • the long CDR H3 of REGN13880 engages the DE loop (E324, V325, T326, D327, N328) and HI loop (D665, N668, S669, F670, I671, T672) of VP3- 5a, with CDR L1 making additional contacts with VP3-5a D665 and N668.
  • CDRs H1, H2, and H3 of REGN13880 also engage VP3-5a by contacting a patch of residues between spanning VR-I (N254, I260, N262, Y274).
  • CDR H1 contacts a single residue in VR-VII of VP3-5a (R550).
  • the binding is further stabilized by CDRs L1 and L3 of REGN13880 contacting VP3-5b VR-VII (Q546, G547, G549) and an additional residue, E718.
  • Cryo-EM structure of AAV2 HBM in complex with REGN13882 [001184] A 2.91 ⁇ cryo-EM structure of AAV2 HBM in complex with the anti-AAV antibody Fab REGN13882 was obtained.
  • REGN13882 binds to the region surrounding the 5-fold symmetry pore and the binding site is located at the boundary of two immediately adjacent VP3 protomers at each 5-fold pore of AAV2 HBM.
  • REGN13882 only contacts two VP3 protomers instead of three. To distinguish between the five VP3 protomers surrounding the 5-fold pore were designated asVP3-5a, VP3-5b, VP3- 5c, VP3-5d, and VP3-5e in a counterclockwise direction.
  • REGN13882 residues contacting VP3 residues of AAV2 HBM are summarized in Table 24. The binding of REGN13882 is primarily mediated by its heavy chain contacting VP3-5a and VP3-5b and light chain contacting VP3-5b.
  • the elongated CDR H3 of REGN 13882 engages the DE (V323, T324, Q325) and HI loops (A655, A667, S668, F669, I670, T671) of VP3-5a.
  • CDRs H1, H2, and H3 of REGN13882 also engage VP3-5a by contacting a patch of residues on the N-terminal side of VR-1 (N254, L256, Q263, S264), with additional contacts made between VR-1 (Q263) and the framework region (FR) of the heavy chain.
  • the binding is further stabilized by CDRs L1, L3, H2, and H3 of REGN13880 contacting VP3-5b VR-II (G328), VR-VII (G546, E548) and two additional residues (T716, N717).
  • the epitope of REGN13882 is primarily located at conserved regions of VP3, allowing this Fab to bind other AAV serotypes including AAV8 and AAV9 as described below.
  • [001185] Cryo-EM structure of AAV8 in complex with REGN13882
  • a 3.10 ⁇ cryo-EM structure of AAV8 in complex with the anti-AAV antibody Fab REGN13882 was obtained.
  • the Fab binds to the region surrounding the 5-fold symmetry pore of AAV8.
  • REGN13882 interacts with AAV8 analogous to how the Fab engages AAV2 HBM.
  • the names used to distinguish between the five VP3 protomers around the 5-fold pore follows the same naming convention used in the AAV2-HBM-REGN13882 structure.
  • REGN13882 residues contacting VP3 residues of AAV8 are summarized in Table 25.
  • the binding of REGN13882 is primarily mediated by heavy chain contacts with VP3-5a and VP3-5b, and by additional contacts of the light chain with VP3-5b.
  • the long CDR H3 of REGN13882 engages the DE (E325, V326, T327, Q328) and HI loops (N670, S671, F672, I673, T674) of VP3-5a.
  • CDRs H2 and H3 of REGN13882 also engage VP3-5a by contacting a patch of residues on the N-terminal side of VR-I (N255, L257, S266).
  • the binding of REGN13882 is further stabilized by CDR L3 contacting VP3-5b VR-VII (N549) and CDRs H2 and H3 engaging two additional residues, T719 and E720.
  • CDR H3 of REGN13882 engages the DE loop (E324, V325, T326, D327) and HI loop (N668, S669, F670, T672) of VP3-5a.
  • CDRs H1, H2, and H3 of REGN13880 also engage VP3-5a by contacting a patch of residues on the N-terminal end of VR-I (N254, L256, I260, N262).
  • CDR H1 contacts a single residue in VR-VII of VP3-5a (D551).
  • Table 18 shows a summary of cryo-EM data collection and processing for the present Example.
  • Table 18 shows a summary of REGN13878 residues contacting AAV2 HBM VP3 residues. ⁇ Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non-hydrogen atoms of antibody. Residues corresponding to REGN13878 interactions with VP3 protomers from the adjacent trimer core #2, as described in the text, are italicized.
  • Table 20 shows a summary of ⁇ summary of REGN13883 residues contacting AAV2 HBM VP3 residues. Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non-hydrogen atoms of antibody. ⁇ Residues corresponding to ⁇ ⁇ Attorney Docket No.250298.000954 REGN13883 interactions with VP3 protomers from the adjacent trimer core #2, as described in the text, are italicized. Table 20.
  • Table 21 shows ⁇ a summary of REGN13880 residues contacting AAV2 HBM VP3 residues. ⁇ Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non-hydrogen atoms of antibody. Table 21. Summary of REGN13880 Residues Contacting AAV2 HBM VP3 Residues ⁇ ⁇ Attorney Docket No.250298.000954 [001193] Table 22 shows a summary of REGN13880 residues contacting AAV8 VP3 residues.
  • Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non-hydrogen atoms of antibody. ⁇ ⁇ Attorney Docket No.250298.000954 Table 22. Summary of REGN13880 Residues Contacting AAV8 VP3 Residues ⁇ ⁇ Attorney Docket No.250298.000954 [001194] Table 23 shows a summary of REGN13880 residues contacting AAV9 VP3 residues. ⁇ Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non-hydrogen atoms of antibody. Table 23.
  • Table 24 shows a ⁇ summary of REGN13882 residues contacting AAV2 HBM VP3 residues. Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non-hydrogen atoms of antibody. Table 24. Summary of REGN13882 Residues Contacting AAV2 HBM VP3 Residues ⁇ [001196] Table 25 shows summary of REGN13882 residues contacting AAV8 VP3 residues.
  • Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non- hydrogen atoms of antibody. ⁇ ⁇ Attorney Docket No.250298.000954 Table 25. Summary of REGN13882 Residues Contacting AAV8 VP3 Residues ⁇ [001197] Table 26 shows a summary of REGN13882 residues contacting AAV9 VP3 residues. ⁇ Contacting residues are defined as amino acids with non-hydrogen atoms within 4 ⁇ of non-hydrogen atoms of antibody. Table 26. Summary of REGN13882 Residues Contacting AAV9 VP3 Residues ⁇ ⁇ Attorney Docket No.250298.000954 Example 5.
  • Anti-AAV x anti-ASGR1 Alternative Format Design and Binding Enzyme- linked immunosorbent assay (ELISA) [001198] A schematic representation of alternative format antibody designs for anti-AAV x anti-ASGR1 bispecific antibodies is shown in Figure 3. A description of corresponding alternative format antibody molecules is displayed in Figure 4. As illustrated in Figure 5, IgG1 versions of anti-AAV mAbs comprising binding arms of REGN13072 (4059311_ULC1- 39_38741_MS) and REGN13221 (4059310_ULC1-39_51231_MS) were selected for inclusion in the binding ELISA described herein based on their respective abilities to enhance internalization of AAV2 expressing NanoLuc genomes into FcGR1-expressing HEK 293T cells.
  • Anti-AAV mAb REGN13072 (4059311_ULC1-39_38741_MS) was functional at low antibody concentrations (1 VG:4 mAb) and did not require saturated antibody binding to the AAV.
  • REGN13221 (4059310_ULC1-39_51231_MS) was not neutralizing when in excess concentrations (1 VG:10K mAb) and required saturated antibody binding to the AAV.
  • Table 27 shows a description of identifiers for the anti-AAV antibodies of the present Example. Table 27.
  • Anti-AAV Antibody Identifiers [001200] Anti-AAV mAbs of the present Example (4059311_ULC1-39_38741_MS, 4059310_ULC1-39_51231_MS) were assessed for binding to AAV serotype AAV2 HBM. ⁇ ⁇ Attorney Docket No.250298.000954 Viral particles were first diluted in PBS and used to coat black 96-well Maxisorp plates (2x10 8 VGs/well) overnight at 4°C. The plates were then emptied, pat-dried, washed 4x with KPL 1x Wash Buffer, and incubated with 3%-BSA blocking buffer overnight at 4°C.
  • Alternative format antibodies were diluted 1:3 in ADB from a starting concentration of 1 ⁇ g/ml.
  • the blocked plates were then emptied, pat-dried, washed 4x, and incubated with alternative format antibodies for 1 hour at RT.
  • the plates were once again emptied, pat- dried washed 4x, and then incubated with goat anti-human HRP secondary antibody (or goat anti-mouse HRP secondary for AAV2 A20) at a 1:10,000 dilution in ADB for 1 hour at RT.
  • the plates were emptied, pat-dried, washed 6x, followed by addition of SuperSignal ELISA Pico Chemiluminescent substrate.
  • Luminescence was detected using a plate reader (Tecan Infinite M200 Pro) and analyzed using GraphPad Prism. Graphs showing binding of 36 alternative format antibodies to AAV2 HBM relative to controls across various antibody concentrations are displayed in Figures 6A-6J.
  • Alternative format antibodies have AAV binding efficacy similar to other antibodies with the same binding arms.
  • Alternative format antibodies with 4059311_ULC1-39_38741_MS binding arms required less antibody for saturated binding as compared to alternative format antibodies with 4059310_ULC1- 39_51231_MS binding arms.
  • Example 6 Example 6
  • Bispecific antibodies can effectively retarget adeno-associated virus (AAV) to the central nervous system (CNS) [001201]
  • the present Example was designed to test retargeting of adeno-associated virus (AAV) to the central nervous system (CNS) using bispecific antibodies.
  • Anti-AAV antibodies that bind, e.g., to AAV capsid epitopes, were formatted in bispecific formats for retargeting of AAV to the central nervous system (CNS).
  • An exemplary representation of an anti-transferrin receptor (TfR) x anti-AAV bispecific antibody retargeting AAV to the CNS, including crossing of the blood-brain barrier (BBB), is schematized in Figure 7A.
  • FIG. 7B A comparison of wild-type (WT) AAV9, covalent-platform control and bispecific platform retargeting of AAV to cerebellum, cortex, and hippocampus assessed using histochemical staining approaches is shown in Figure 7B. Liver detargeting could also be controlled by modulating antibody concentration [Ab] and/or use of a detargeting capsid (see, e.g., Figures 8A-8B).
  • a comparison of transduction efficiency between the covalent platform and the bispecific platform using cDNA quantitation is shown in Figure 8A. The results showed that the bispecific platform achieved comparable efficacy to the covalent platform. Higher antibody concentrations achieved partial liver detargeting with WT AAV9. Further detargeting of AAV was achieved when paired with a detargeted AAV.
  • a comparison between detargeted AAV9 and WT AA9 using the bispecific platform in cerebellum and liver is shown in Figure 8B.
  • ⁇ ⁇ Attorney Docket No.250298.000954 These data support fine tuning of AAV retargeting and detargeting by modulating capsid and antibody concentration to achieve desired in vivo tropism.
  • Example 7. Transduction assay to assess the efficacy of anti-AAV x anti-CACNG1 bispecific antibodies
  • Adeno-associated viruses are members of the Parvovirus family of non- enveloped single-stranded DNA viruses.
  • the AAV capsid structure comprises three viral proteins, VP1, VP2, and VP3, that are encoded from a single open reading frame.
  • An icosahedral capsid is formed from the assembly of 60 monomers of VP1, VP2, and VP3 in relative amounts of 1:1:10.
  • the AAV capsid contains residues critical for mediating cell and receptor binding.
  • the capsids of many serotypes bind to the adeno-associated virus receptor (AAVR), which is required for efficient transport of particles to the trans-golgi network, an essential step of the viral transduction pathway.
  • AAVR adeno-associated virus receptor
  • Antibodies that bind to regions of the capsid attributed to either glycan or AAVR binding have been shown to neutralize AAV transduction. It has also been shown that antibodies can be used to bridge AAV capsids to target cells to enhance transduction efficiency.
  • Anti-AAV x anti-CACNG1 bispecific antibodies of the present disclosure (AF70: anti-AAV#70 scFv fused to N-terminal of anti-CACNG1 REGN10717 hIgG1 N297G antibody; AF71: anti-AAV#70 scFv fused to C-terminal of anti-CACNG1 REGN10717 hIgG1 N297G antibody; Figure 9, Figures 30A-30G, and Table 30) were assessed for their capacity to enhance internalization of AAV Hu37 particles expressing green fluorescent protein (GFP) genome into mouse CACNG1-expressing HEK 293 cells and human CACNG1-expressing HEK 293 cells.
  • GFP green fluorescent protein
  • both cell lines were seeded in a clear wall clear bottom 96-well plate at 10,000 cells per well using DMEM (supplemented with 10% FBS, MEM NEAA, Pen Strep) and incubated at 37°C and 5% CO 2 .
  • DMEM supplied with 10% FBS, MEM NEAA, Pen Strep
  • AF70 and AF71 antibodies were first diluted in 1xPBS and then serially diluted (3-fold) in 1xPBS to obtain the appropriate viral genome to antibody molar ratios (1:1, 1:3, 1:9, 1:27, and 1:81).
  • AAV Hu37 and control virus (1/8 Spytagged AAV9 W503A conjugated with anti-CACNG1 bivalent antibody) were diluted in 1xPBS + 0.001% Pluronic.
  • Equal volumes of AAV Hu37 and antibody dilutions were combined to obtain the above viral genome to antibody molar ratios and incubated for 1 hour at 37°C and 5% CO 2 . After a 1-hour incubation, the appropriate volumes of the viral genome to antibody complexes were added to the cells to yield an MOI of 2E+04 and 2E+05 VG per cell.60-hours post transduction, the transduced cells were analyzed by flow cytometry. To prepare samples for flow cytometry, the supplemented media was discarded, and the cells were washed once with 100 L 1xPBS.
  • C2C12 myoblasts were seeded on collagen I coated plates with black walls at 10,000 cells per well; human skeletal muscle derived myoblasts were seeded on collagen I coated plates with black walls at 12,500 cells/well. After 24 hours, human myoblasts were treated with differentiation media (Cook Myosite, MD-5555) for 3 days, and the C2C12 myoblasts were treated with DMEM + 2% horse serum for 2 days.
  • differentiation media Cook Myosite, MD-5555
  • the cells were treated with the AAVhu37 and antibody complexes (1E5 viral genomes per cell) as described above.
  • VVT885 (1/8 Spytagged AAV9 W503A conjugated to anti-CACNG1 antibody) diluted in PBS was used as a positive control.
  • the cells were fixed with 4% PFA for 15 minutes, washed twice with 1xPBS and blocked with 20% goat serum + 0.3% Triton X-100. The wells were incubated with MF-20 (DSHB) overnight in blocking buffer at 1:100 to stain for Myosin Heavy Chain (MyHC).
  • MF-20 DSHB
  • Transduction efficiency was optimal between molar ratios of 1 AAV to 1 antibody and 1 AAV to 9 antibodies as assessed by %GFP positive cells ( Figure 10 and Figure 12) and mean fluorescence intensity ( Figure 11 and Figure 13).
  • incubation of AAV with AF70 and AF71 enhanced transduction into differentiated C2C12 myotobes ( Figure 14 and Figures 16A-16D) and differentiated human myotubes ( Figure 15 and Figures 17A-17D) as determined by quantification of GFP positive cells ( Figures 16A-16D and Figures 17A- 17D).
  • C2C12 was most efficiently transduced with AAV complexed with AF71 at molar ratio 1:3 and 1:9 ( Figure 14).
  • Peak transduction efficacy of AAV retargeted with bispecific antibodies was comparable to that of control AAV covalently conjugated to an anti-CACNG1 ⁇ ⁇ Attorney Docket No.250298.000954 antibody (VVT885), except for AF70 on human myotubes which showed efficacy but of lesser magnitude.
  • the present Example was designed to test the binding of anti-AAV x anti-mTfR alternative format (AF) antibodies (Abs) to AAV9W503A virus by enzyme-linked immunoassay (ELISA).
  • ELISA enzyme-linked immunoassay
  • alternative format (AF) antibodies primary antibodies
  • ADB assay diluent buffer
  • AF alternative format
  • ID 1-13 A description the anti-AAV x anti-mTfR alternative format (AF) antibodies tested in the present example (ID 1-13) is shown in Figures 18A-18B (see also, e.g., Figures 29A-29I and Table 30).
  • REGN1932 anti-FelD1
  • REGN13072 bivalent with NGS/MS 70
  • REGN13221 bivalent with NGS/MS 64
  • Additional control conditions were ⁇ AAV9W503A only and AAV9W503A + secondary antibody only. ⁇ Primary antibody dilutions were then transferred to the coated plates and incubated for 1 hour at RT. ⁇ For the secondary antibody, an ⁇ -Human-HRP at 1:10,000 dilution in ADB buffer at RT was used.1xKPL buffer was used for in-between step washes. The SuperSignal ELISA Pico Chemiluminescent Substrate Kit was used for development. The substrate was added and incubated for ⁇ 1 minute at RT and then the readout was performed using the Tecan Infinite m200 Pro instrument.
  • the present Example tested retargeting of AAV9W503A using mouse TfR (mTfR) alternative format antibodies (see, e.g., Figures 18A-18B, Figures 29A-29I, and Table 30) on 293T cells expressing the mTfR receptor.
  • mTfR mouse TfR
  • the multiplicity of infection (MOI) was 1x10 5 per well and the total vector genomes (VGs) was 1x10 9 per well.
  • Alternative format antibodies were diluted in 1XPBS and 4-fold serial dilutions were performed to attain various AAV:Antibody ratios from a starting AAV:Antibody ratio of 1:10,0000.
  • the starting AAV:Antibody ratio was 1:7750 (low stock concentration).
  • the virus and the antibodies were mixed and incubated for 1 hour at 37°C.
  • the Virus:Antibody complex was then added to the 293T cells expressing the mTfR receptor and incubated for 72 hours at 37°C.
  • a Firefly luciferase (FLuc) read-out protocol was performed to measure the luciferase signal.
  • FLuc Firefly luciferase
  • the plate was spun at 1500 RPM for 1 minute and then the supernatant was discarded.50 l lysis buffer was added per well (Glo- Lysis, Promega E2661) for 5 minutes. Then, 100 l luciferase substrate was added per well and incubated for approximately 1 minute. The plate was analyzed using PerkinElmer2030 (or SpectraMax) with a setting selection of “Luminescence Corning Black Clear Bottom”. The wells to be read and measured were selected and following the reading/measurement, data were exported.
  • the AAV9W503A background value was 0.
  • the results showed that anti- AAV x anti-mTfR alternative format (AF) antibodies with anti-AAV NGS/MS #70 retarget AAV9W503A at low ratios and have higher peak when compared to anti-AAV x anti- mTfR AF Abs w/ anti-AAV NGS/MS #64 ( Figure 22).
  • AF alternative format
  • AAV9 Retargeting assay using AAV9 and an eGFP reporter
  • mTfR mouse TfR
  • AF1 and AF7 see, e.g., Figures 18A-18B and Table 30
  • AAV9 was diluted in 1xPBS+0.001% Pluronic.
  • MOI multiplicity of infection
  • Alternative format antibodies were diluted in 1XPBS and 3-fold serial dilutions were performed to attain various AAV:Antibody ratios from a starting AAV:Antibody ratio of 1:729.
  • the virus and the antibodies were mixed and incubated for 1 hour at 37°C.
  • the AAV:Antibody complex was then added to the 293T cells expressing the mTfR receptor and ⁇ ⁇ Attorney Docket No.250298.000954 incubated for 72 hours at 37°C. After the 72-hour incubation period, a GFP read-out protocol was performed to measure fluorescence. [001210] To prepare samples for flow cytometry, supplemented media was discarded, and cells were washed once with 100 L 1xPBS.

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Abstract

L'invention concerne des anticorps anti-virus adéno-associés (AAV) et des fragments d'antigène de ceux-ci, ainsi que des anticorps multispécifiques (par exemple, des anticorps bispécifiques) qui se lient à la fois à une capside d'une particule d'AAV et à une molécule sur une surface cellulaire, des complexes moléculaires associés, des matrices d'affinité et des compositions pharmaceutiques, et leurs procédés d'utilisation.
PCT/US2025/032630 2024-06-07 2025-06-06 Molécules de liaison à l'antigène qui se lient à des particules d'aav et leurs utilisations Pending WO2025255452A2 (fr)

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