EP4746902A2 - Zusammensetzungen zur abgabe von plasmodium-antigenen und zugehörige verfahren - Google Patents

Zusammensetzungen zur abgabe von plasmodium-antigenen und zugehörige verfahren

Info

Publication number: EP4746902A2
Authority: EP; European Patent Office
Prior art keywords: seq; plasmodium; polypeptide; amino add; amino
Prior art date: 2023-07-21
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.): Pending

Application number

EP24754502.3A

Other languages

English (en)

French (fr)

Inventor

Adam ZUIANI

Charles Lefco DULBERGER

Asaf PORAN

Daniel Abram Rothenberg

John SROUJI

Charles JENNISON

Patricia DOS SANTOS MEIRELES

Anja DOKIC

Annette VOGEL

Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

BioNTech SE

Original Assignee

BioNTech SE

Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)

2023-07-21

Filing date

2024-07-19

Publication date

2026-05-27

2024-07-19 Application filed by BioNTech SE filed Critical BioNTech SE

2026-05-27 Publication of EP4746902A2 publication Critical patent/EP4746902A2/de

Status Pending legal-status Critical Current

Links

Classifications

- 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
- A61K39/002—Protozoa antigens
- A61K39/015—Hemosporidia antigens, e.g. Plasmodium antigens
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
- A61P33/06—Antimalarials
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/44—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from protozoa
- C07K14/445—Plasmodium
- 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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/53—DNA (RNA) vaccination
- 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
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55555—Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

Malaria is a mosquito-borne infectious disease caused by protozoan parasites of the Plasmodium genus.
Plasmodium antigens also referred to herein as "malaria antigens”.
a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof.
a portion is an antigenic portion.
a polyribonucleotide as provided herein is an isolated polyribonucleotide. In some embodiments, a polyribonucleotide as provided herein is an engineered polyribonucleotide. In some embodiments, a polyribonucleotide as provided herein is a codon-optimized polyribonucleotide.
one or more Plasmodium Rh5 invasion complex antigens comprise: (i) one or more Plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof; (ii) one or more Plasmodium Cysteine-Rich Protective Antigen (CyRPA) polypeptides or antigenic portions thereof; (iii) one or more Plasmodium Rh5-interacting Protein (Ripr) polypeptides or antigenic portions thereof; (iv) one or more Plasmodium P113 polypeptides or antigenic portions thereof; (v) one or more Plasmodium thrombospondin-related apical merozoite protein (TRAMP) polypeptides or antigenic portions thereof; or (vi) one or more Plasmodium cysteine-rich small secreted protein (CSS) polypeptides or antigenic portions thereof.
Rh5 Plasmodium reticulocyte-binding protein homolog 5
CyRPA Plasmodium Cysteine-R
a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof. In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises one or more Plasmodium reticulocytebinding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof.
Rh5 Plasmodium reticulocytebinding protein homolog 5
one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise one or more antigenic portions of Plasmodium Rh5.
one or more Plasmodium Rh5 antigenic portions comprise one or more ordered domains of Rh5.
one or more Plasmodium Rh5 antigenic portions comprise two ordered domains of Rh5.
two ordered domains of Rh5 are directly adjacent to one another.
one or more Plasmodium Rh5 antigenic portions comprise a cysteine at position 203, 329, or both, as numbered according to SEQ ID NO: 1.
one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino acid sequence according to SEQ ID NO: 118, 119, 124, or 126, (ii) an amino acid sequence according to SEQ ID NO: 120 or 128, or (iii) a combination thereof.
one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, 329, or both, as numbered according to SEQ ID NO: 1.
one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino acid sequence according to SEQ ID NO: 121, 122, 125, or 127, (ii) an amino add sequence according to SEQ ID NO: 123 or 129, or (iii) a combination thereof.
one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, as numbered according to SEQ ID NO: 1.
one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 329, as numbered according to SEQ ID NO: 1.
one or more Plasmodium Rh5 antigenic portions comprise one, two, three, or four N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two N-llnked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a substitution of NX[T/S] to QX[T/S] and/or a substitution of NX[T/S] to NXA.
one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino add sequence according to SEQ ID NO: 124, 125, 126, or 127, (ii) an amino add sequence according to SEQ ID NO: 128 or 129, or (ii) a combination thereof.
one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution that prevents glycosylation at position 214, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution that prevents glycosylation at position 297, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
one or more Plasmodium Rh5 antigenic portions comprise a PMX deavage site. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence according to SEQ ID NO: 118, 121, 124, or 125. In some embodiments, one or more Plasmodium Rh5 antigenic portions do not comprise a PMX cleavage site.
one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence according to SEQ ID NO: 119, 122, 126, or 127.
one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 130, (ii) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127, (iii) an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 131, (iv) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 120, 123, 128, or 129, or (v) a combination thereof.
one or more Plasmodium Rh5 antigenic portions comprise (i) an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127, (ii) an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 120, 123, 128, or 129, or (iii) a combination thereof.
one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino add sequence according to SEQ ID NO: 121 or 122, and (ii) an amino add sequence according to SEQ ID NO: 120.
one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino acid sequence according to SEQ ID NO: 125 or 127, and (ii) an amino add sequence according to SEQ ID NO: 120 or 128.
one or more Plasmodium Rh5 antigenic portions comprise an amino acid sequence with at least 85% sequence identity to an amino add sequence of positions 26-363 of SEQ ID NO: 94, 95 or 99. In some embodiments, one or more PlasmotSum Rh5 antigenic portions comprise an amino acid sequence with at least 85% sequence identity to an amino add sequence of positions 26-526 of SEQ ID NO: 89. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence of positions 26-363 of SEQ ID NO: 94. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence of positions 26-363 of SEQ ID NO: 95.
one or more Plasmodium Rh5 antigenic portions comprise an amino acid sequence of positions 26-363 of SEQ ID NO: 99. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence of positions 26-526 of SEQ ID NO: 89. In some embodiments, one or more Plasmodium Rh5 antigenic portions do not comprise a disordered domain of Rh5. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a PMX cleavage site. In some embodiments, a PMX deavage site comprises or consists of an amino acid sequence of NFLQ.
one or more Plasmodium Rh5 Invasion complex antigens comprise a Plasmodium CyRPA polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 Invasion complex antigens comprise an antigenic portion of Piasmocfium CyRPA.
a Plasmodium CyRPA antigenic portion comprises an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 169. In some embodiments, a Plasmodium antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 169.
a Plasmodium CyRPA antigenic portion comprises one, two, or three N-llnked glycosylation sites. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a substitution of NX[T/S] to NXA. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium CyRPA antigenic portion comprises an asparagine at position 145, position 322, position 338, or a combination thereof, as numbered according to SEQ ID NO: 3.
a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, position 338, or a combination thereof, as numbered according to SEQ ID NO: 3.
a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, and position 338, as numbered according to SEQ ID NO: 3.
a Plasmodium CyRPA antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 134.
a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 170.
one or more Plasmodium Rh5 invasion complex antigens comprise a Plasmodium Pl 13 polypeptide or antigenic portion thereof.
one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium Pl 13.
a Plasmodium P113 antigenic portion comprises an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 135. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 135.
a Plasmodium P113 antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a Plasmodium P113 antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium P113 antigenic portion comprises a substitution of NX[T/S] to QX[T/S], In some embodiments, a Plasmodium P113 antigenic portion comprises a substitution of NX[T/S] to NXA. In some embodiments, a Plasmodium P113 antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
a Plasmodium P113 antigenic portion comprises an asparagine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.
a Plasmodium Pl 13 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.
a Plasmodium Pl 13 antigenic portion comprises a glutamine position 207, position 268, position 317, position 360, position 661, position 697, position 779, and position 876, as numbered according to SEQ ID NO: 6.
a Plasmodium P113 antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 136.
one or more Plasmodium Rh5 invasion complex antigens comprise a Plasmodium Rlpr polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium Rlpr.
a Plasmodium Rlpr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 178. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 177. In some embedments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 177. [0025] In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 176.
a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 175. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 175. [0026] In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 174. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 174.
a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 190.
a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 171. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 171.
a Plasmodium Ripr antigenic portion comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve N-llnked glycosylation sites.
a Plasmodium Ripr antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium Ripr antigenic portion comprises a substitution of NX[T/S] to QX[T/S].
a Plasmodium Ripr antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
a Plasmodium Ripr antigenic portion comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 190.
a Plasmodium Ripr antigenic portion comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.
a Plasmodium Ripr antigenic portion comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.
a Plasmodium Ripr antigenic portion comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, as numbered according to SEQ ID NO: 2.
a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 171.
a Plasmodium Ripr antigenic portion comprises a PMX cleavage site.
a PMX deavage site comprises or consists of an amino acid sequence of SMLE.
one or more Plasmodium Rh5 invasion complex antigens comprise a Plasmodium TRAMP polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium TRAMP.
a Plasmodium TRAMP antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 179.
a PfasmodiumTRMAP antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 179.
a Plasmodium TRAMP antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites.
a Plasmodium TRAMP antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
a Plasmodium TRAMP antigenic portion comprises a substitution of NX[T/S] to QX[T/S],
a / ⁇ asmoa6i/fl)TRAMP antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a PiasmodmmTRMAP antigenic portion comprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4.
a Plasmodium TRAMP antigenic portion comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 181.
a Plasmodium TRAMP antigenic portion comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4.
a Plasmodium TRAMP antigenic portion comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4.
a Plasmodium TRAMP antigenic portion comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 4.
a Plasmodium TRAMP antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 180.
a Plasmodium TRAMP antigenic portion comprises a PMX deavage site.
a PMX deavage site comprises or consists of an amino add sequence of HFLQ.
a Plasmodium TRAMP antigenic portion comprises a SUB2 cleavage site.
a SUB2 deavage site comprises or consists of an amino add sequence according to SEQ ID NO: 193.
one or more Plasmodium Rh5 invasion complex antigens comprise a Ptasmodum CSS polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium CSS.
a Plasmodium CSS antigenic portion comprises one or more cysteine to serine mutations.
a Ptasmodum CSS antigenic portion comprises a C30S mutation, a C80S mutation, a C276S mutation, or a combination thereof.
a Plasmodium CSS antigenic portion comprises a serine at position 30 and position 80, as numbered according to SEQ ID NO: 5 or 214.
a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS antigenic portion comprises a serine at position 30 and position 276, as numbered according to SEQ ID NO: 5 or 214.
a Plasmodium CSS antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 195.
a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 182.
a Plasmodium CSS antigenic portion comprises one, two, three, four, five, or six N-inked glycosylation sites. In some embodiments, a Plasmodium CSS antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CSS antigenic portion comprises a substitution of NX[T/S] to QX[T/S], In some embodiments, a Plasmodium CSS antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium CSS antigenic portion comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, a Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 214.
a Plasmodium CSS antigenic portion comprises a glutamine at position 74 and position 88, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 184.
a Plasmodium CSS antigenic portion comprises an asparagine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.
a Plasmodium CSS antigenic portion comprises a glutamine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.
a Plasmodium CSS antigenic portion comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 195.
a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 185.
a Plasmodium CSS antigenic portion comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.
a Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.
a Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182.
a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 183.
a polypeptide comprises two or more Piasmodum Rh5 invasion complex polypeptides or antigenic portions thereof.
Rh5 invasion complex polypeptides or antigenic portions thereof comprise two selected from (I) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, (ii) one or more Plasmodium C'fRPk polypeptides or antigenic portions thereof, (iii) one or more Plasmodium Ripr polypeptides or antigenic portions thereof, (iv) one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof, (v) one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, and (vi) one or more Plasmodium CSS polypeptides or antigenic portions thereof.
a polypeptide comprises one or more Plasmodium Rh5 antigenic portions and further comprises: (i) one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, (ii) one or more Plasmodium Ripr polypeptides or antigenic portions thereof, (iii) one or more Plasmodium P113 polypeptides or antigenic portions thereof, (iv) one or more Ptis/nodt/m TRAMP polypeptides or antigenic portions thereof, or (v) one or more Plasmodium CSS polypeptides or antigenic portions thereof.
a polypeptide comprises one or more Plasmodium Rh5 antigenic portions, wherein the polypeptide comprises: (i) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more Plasmodium CyRPA polypeptides or antigenic portions thereof; (il) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof; (iii) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium Ripr polypeptides or antigenic portions thereof; (hr) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium TRAMP polypeptides or antigenic portions thereof; or (v) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium CSS polypeptides or antigenic portions thereof.
a polypeptide comprises a secretory signal.
a secretory slgial is located at the N-terminus of the polypeptide.
a secretory signal is a heterologous secretory signal.
a heterologous secretory signal comprises or consists of a viral secretory signal.
a viral secretory signal comprises or consists of an HSV secretory signal.
an HSV secretory signal comprises or consists of an HSV-1 or HSV-2 secretory signal.
an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal.
gD secretory signal consists of an amino add sequence according to SEQ ID NO: 42.
a secretory signal comprises or consists of a Plasmodium seaetarf signal.
a secretory signal comprises or consists of a Plasmodium CyRPA secretory signal.
a secretory signal comprises or consists of a Plasmodium Ripr secretory signal.
a secretory signal comprises or consists of a Plasmodium CSS secretory signal.
a secretory signal comprises or consists of an amino add sequence according to SEQ ID NOs: 186, 187, 188.
a polypeptide comprises a transmembrane region.
a transmembrane region comprises a heterologous transmembrane region.
a transmembrane region comprises or consists of a Plasmodium transmembrane region. In some embodiments, a transmembrane region comprises or consists of a P113 Plasmodium transmembrane region. In some embodiments, a transmembrane region comprises or consists of a TRAMP Plasmodium transmembrane region.
a heterologous transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, an HSV transmembrane region comprises or consists of an HSV- 1 or HSV-2 transmembrane region. In some embodiments, an HSV transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, an HSV gD transmembrane region consists of an amino add sequence according to SEQ ID NO: 75.
a polypeptide does not comprise a transmembrane region.
a polypeptide comprises a multimerization domain.
a multimerization domain is a trimerization domain.
a trimerization domain is a C -terminal domain of T4 fibritin (e.g., foldon domain).
a foldon domain comprises or consists of an amino acid sequence according to SEQ ID NO: 78.
a polypeptide comprises a self-assembling nanoparticle domain.
a self-assembling nanopartide domain is a ferritin domain.
a ferritin domain is from H. pylori.
a ferritin domain comprises or consists of a sequence according to SEQ ID NO. 88.
a polypeptide comprises one or more linkers. In some embodiments, a polypeptide comprises one or more glydne-serine linkers. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 80. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 83. In some embodiments, one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 84. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 86. In some embodiments, one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 137. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 138.
a polypeptide comprises a linker after an amino add sequence of the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof.
one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof are one or more P. faidparum Rh5 invasion complex polypeptides or antigenic portions thereof.
one or more Plasmodium Rh5 polypeptides or antigenic portions thereof are one or more P. falciparum Rh5 antigenic portions.
one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof are a P. falciparum CyRPA polypeptide or antigenic portion thereof.
one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. faidparum Pl 13 polypeptide or antigenic portion thereof.
one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. falciparum Ripr polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparum TRAMP polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparum CSS polypeptide or antigenic portion thereof.
polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises a secretory signal, and two Plasmodium Rh5 ordered domains.
a polypeptide comprises (i) a secretory signal, (II) two Plasmodium Rh5 ordered domains, (iH) a linker, and (iv) a multimerization domain.
a polypeptide comprises (I) a secretory signal, (II) two Plasmodium Rh5 ordered domains, (ill) a linker, and (iv) a transmembrane region.
a polypeptide comprises (i) a secretory signal, (ii) two Plasmodium Rh5 ordered domains, (iii) a linker, and (iv) a self-assembling nanoparticle domain.
a secretory signal is a viral secretory signal.
a secretory skyial is an HSV glycoprotein D (gD) secretory signal, optionally wherein the HSV gD secretory signal consists of an amino acid sequence according to SEQ ID NO: 42.
a polypeptide does not comprise a Plasmodium Rh5 disordered domain. In some embodiments, two Plasmodium Rh5 ordered domains are directly adjacent.
a polypeptide comprises a C203Y substitution, as numbered according to SEQ ID NO: 1
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 94. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 94.
a linker comprises or consists of a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86.
a multimerization domain Is a C-terminal domain of T4 fibrttin (e.g., fokion domain).
a foldon domain comprises or consists of an amino acid sequence according to SEQ ID NO: 78.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 90. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 90.
a transmembrane region is a heterologous transmembrane region.
a transmembrane region is a viral transmembrane region.
a viral transmembrane region Is a HSV gD transmembrane region.
a HSV gD transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 92. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 92.
a linker comprises or consists of a glycine-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 137.
a self-assembling nanoparticle domain is a ferritin domain.
a ferritin domain comprises or consists of a sequence according to SEQ ID NO: 88.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 96. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 96.
Plasmodium Rh5 ordered domains comprise one or more N-linked glycosylation sites.
Plasmodium Rh5 ordered domains comprise one NX[T/S] to QX[T/S] substitution, optionally wherein the substitution is a N214Q substitution, as numbered according to SEQ ID NO: 1.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 99. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 99.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 98. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 98.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 101. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 101.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 100. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 100.
Plasmodium Rh5 ordered domains comprise two NX[T/S] to QX[T/S] substitutions, optionally wherein the two substitutions comprise N214Q and N297Q substitutions.
Plasmodium Rh5 ordered domains comprise an amino add substitution at al of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 95. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 95.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 91. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 91.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 93. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 93.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 97. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 97.
polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises (i) a secretory signal, (ii) a Plasmodium Rh5 N-terminal disordered domain, (iii) a Plasmodium Rh5 ordered domain, (iv) a Plasmodium Rh5 linking disordered domain, and (v) a Plasmodium Rh5 C-terminal ordered domain.
a secretory signal Is a heterologous secretory signal.
a heterologous secretory signal is a viral secretory signal.
a viral secretor signal is an HSV glycoprotein D (gD) secretory signal.
gD HSV glycoprotein D
an HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 42.
a polypeptide comprises one or more N-linked glycosylation sites. In some embodiments, a polypeptide comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation, optionally wherein the polypeptide comprises an amino add substitution at all of the N-linked glycosylation sites. [0093] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 89. In some embodiments, a polypeptide is or comprises an amino add sequence according to SEQ ID NO: 89.
the present disdosure further provides a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises (i) a secretory signal, and (ii) a Plasmodium CyRPA polypeptide or antigenic portion thereof.
a polypeptide comprises (i) a secretory signal, (ii) a Plasmodium CyRPA polypeptide or antigenic portion thereof, (Si) a linker, and (iv) a multimerization domain.
a polypeptide comprises (i) a secretory signal, (ii) a Plasmodium CyRPA polypeptide or antigenic portion thereof, and (iii) a transmembrane region.
a polypeptide comprises (i) a seaetory signal, (II) a Plasmodium Of'RPk polypeptide or antigenic portion thereof, (iii) a linker, and (iv) a transmembrane region.
a secretory signal is a heterologous secretory signal.
a heterologous secretory signal is a viral secretory signal.
a viral secretor signal is an HSV glycoprotein D (gD) secretory signal.
gD HSV glycoprotein D
an HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 42.
a secretory signal is a Plasmodium seatiorf signal.
a Plasmodium secretory signal Is a Plasmodium CyRPA seaetory signal.
a Plasmodium CyRPA secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 187.
a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence Identity to a sequence comprising SEQ ID NO: 133.
a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of SEQ ID NO: 169 or 133. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of amino adds 29-362 of SEQ ID NO: 169 or 133.
a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S] and/or a substitution of NX[T/S] to NXA.
a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises an N145Q, N322Q, N338Q or a combination thereof.
a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises an N145Q, N322Q, N338Q.
a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.
a multimerization domain is a C-terminal domain of T4 fibritin (foldon domain).
a foldon domain comprises or consists of an amino add sequence according to SEQ ID NO: 78.
a transmembrane region is a heterologous transmembrane region.
a transmembrane region is a viral transmembrane region.
a viral transmembrane region is a HSV gD transmembrane region.
a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 140. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 140.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 141. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 141.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 142. In some embodiments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 142.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 143. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 143.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 144. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 144.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 198. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 198.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 199. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 199.
a polyribonucleotide encoring a polypeptide, wherein the polypeptide comprises (I) a secretory signal, (H) a Plasmodium P113 polypeptide or antigenic portion thereof, (ill) a linker, and (Iv) a transmembrane region.
a polypeptide comprises: (I) a secretory signal, (li) a Plasmodium P113 polypeptide or antigenic portion thereof, (Hi) a linker, and (iv) a transmembrane region.
a secretory signal is a heterologous secretory signal.
a heterologous secretory signal is a viral secretory signal.
a viral secretor signal is an HSV glycoprotein D (gD) secretory signal.
gD HSV glycoprotein D
an HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 42.
a Ifriker comprises a glydne-serine linker.
a linker comprises or consists of an amino add sequence according to SEQ ID NO: 86.
a transmembrane region is a heterologous transmembrane region.
a transmembrane region is a viral transmembrane region.
a viral transmembrane region is a HSV gD transmembrane region.
a HSV gD transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.
a Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 135. In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 135.
a Plasmodium P113 polypeptide or antigenic portion thereof comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises one or more substitutions of NX[T/S] to QX[T/S] and/or substitutions of NX[T/S] to NXA.
a Plasmodium P113 antigenic portion comprises an asparagine at position 207, position 268, position 317, position 360, position 661, position 694, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.
a Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 694, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.
a Plasmodium P113 antigenic portion comprises a glutamine position 207, position 268, position 317, position 360, position 661, position 694, position 779, and position 876, as numbered according to SEQ ID NO: 6.
Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 136.
polypeptide comprises (I) a secretory signal, and (H) a Plasmodium Rlpr polypeptide or antigenic portion thereof.
a polypeptide comprises (I) a secretory signal, (II) a Plasmodium Rlpr polypeptide or antigenic portion thereof, and (III) a transmembrane region.
a polypeptide comprises (I) a secretory signal, (II) a Plasmodium Rlpr polypeptide or antigenic portion thereof, (ii) a linker, and (hr) a transmembrane region.
a secretory signal Is a Plasmodium secretory signal.
a Plasmodium secretory signal Is a Plasmodium Rlpr secretory signal.
a Plasmodium Ripr secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 186.
a secretory signal is a viral seaetory signal.
a secretory signal is an HSV glycoprotein D (gD) secretory signal.
gD HSV glycoprotein D
a HSV gD secretory signal comprises or consists of an amino add sequence accorcfing to SEQ ID NO: 42.
a Plasmodium Ripr polypeptide or antigenic portion thereof Ripr comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 171. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 171.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve N-linked glycosylation sites. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 171.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 171.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 171.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 172.
a linker comprises a glycine-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 145. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 145.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 147. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 147.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 149. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 149.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 151. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 151.
a transmembrane region is a heterologous transmembrane region.
a heterologous transmembrane region comprises or consists of a viral transmembrane region.
a heterologous transmembrane region comprises or consists of an HSV transmembrane regon.
a HSV transmembrane region comprises or consists of an HSV gD transmembrane region.
a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 146. In some embodments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 146.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 200. In some embodments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 200.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 148. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 148. [0139] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 201. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 201.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 150. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 150.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 202. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 202.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 152. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 152.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 203. In some embedments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 203.
Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 178.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 177. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 177.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 176. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 176.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 175. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 175.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 174. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 174.
Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 190.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S].
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 173.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 153. In some embodiments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 153.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 204. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 204.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 154. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 154.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 205. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 205.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 155. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 155.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 206. In some emboefiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 206.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 156. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 156.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 207. In some emboefiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 207.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 157. In some emboefiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 157. [0161] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 208. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 208.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 158. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 158.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 209. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 209.
polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises (I) a secretory signal, (H) a Plasmodium TRAMP polypeptide or antigenic portion thereof.
a secretory signal is a viral secretory signal.
a secretory signal is an HSV glycoprotein D (gD) secretory signal.
gD HSV glycoprotein D
a HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 179. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 179.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 179.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 179.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 179.
a Pfasmodft/mTRAMP polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 181.
a PiasmodiumTRNAP polypeptide or antigenic portion thereof comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 179.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 179.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 179.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 180.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a PMX cleavage site.
a PMX deavage site comprises or consists of an amino add sequence of HFLQ.
a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a SUB2 cleavage site.
a SUB2 cleavage site comprises or consists of an amino acid sequence according to SEQ ID NO: 193.
a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 159. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 159.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 160. In some embodiments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 160.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 161. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 161.
Futher provided is a polyribonudeotkie encoding a polypeptide, wherein the polypeptide comprises (I) a secretory signal, and (li) a Plasmodium CSS polypeptide or antigenic portion thereof.
a polypeptide comprises (I) a secretory signal, (ii) a Plasmodium CSS polypeptide or antigenic portion thereof, and (ill) a transmembrane region.
a polypeptide comprises (I) a secretory signal, (il) a Plasmodium CSS polypeptide or antigenic portion thereof, (Hi) a linker, and (iv) a transmembrane region.
a secretory signal is a Plasmodium secretory signal.
a Plasmodium secretory signal Is a Plasmodium CSS secretory signal.
a Plasmodium CSS secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 188.
a secretory signal is a viral secretory signal.
a secretory signal is an HSV glycoprotein D (gD) secretory signal.
gD HSV glycoprotein D
a HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 182.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 183.
a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 162. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 162.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 163. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 163.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 164. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 164.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 194.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 195.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 182.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74 and position 88, as numbered according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 184.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 195.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 185.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 183.
a first linker and/or second linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.
a transmembrane region is a heterologous transmembrane region.
a heterologous transmembrane region comprises or consists of a viral transmembrane region.
a heterologous transmembrane region comprises or consists of an HSV transmembrane region.
a HSV transmembrane region comprises or consists of an HSV gD transmembrane region.
a HSV gD transmembrane region comprises or consists of an amino add sequence accorcSng to SEQ ID NO: 75.
a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 165. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 165. [0198] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 166. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 166.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 167. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 167.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 168. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 168.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 210. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 210.
a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 211. In some embedments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 211.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 212. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 212.
a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 213. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 213.
an RNA construct comprises, In 5' to 3' order (I) a 5' UTR, (li) a polyribonucleotide described herein, (ill) a 3' UTR, and (Iv) a polyA tall sequence.
an RNA construct comprises (i) a 5' UTR that comprises or consists of a modified human alpha-globin 5 -UTR; and (ii) a 3' UTR that comprises or consists of a first sequence from the amino terminal enhancer of split (AES) messenger.
AES amino terminal enhancer of split
an RNA construct comprises a 5' UTR.
a 5' UTR comprises or consists of a modified human alpha-globin 5 -UTR.
a 5' UTR consists of a ribonudeic add sequence according to SEQ ID NO: 111.
an RNA construct comprises a 3' UTR.
a 3' UTR that comprises or consists of a first sequence from the amino terminal enhancer of split (AES) messenger RNA and a second sequence from the mitochondrial encoded 12S ribosomal RNA.
a 3’ UTR consists of a ribonudeic add sequence according to SEQ ID NO: 117.
an RNA construct comprises a polyA tail sequence.
a polyA tail sequence is a split polyA tail sequence.
a split polyA tail sequence consists of a ribonudeic add sequence according to SEQ ID NO: 114.
an RNA construct comprises a 5* cap.
an RNA construct comprises a cap proximal sequence comprising positions +1,
a 5’ cap comprises or consists of m7(3'OMeG)(5’)ppp(5')(2 , OMeAi)pGz, wherein Ai is position +1 of the polyribonucleotide, and Gz is position +2 of the polyribonucleotide.
a cap proximal sequence comprises Ai and Gz of the Capl structure, and a sequence comprising: A3A4U5 (SEQ ID NO: 138) at positions +3, +4 and +5 respectively of the polyribonucleotide.
a polyribonucleotide provided herein indudes modified uridines in place of all uridines.
modified uridines are each Nl-methyl-pseudouridine.
compositions comprising one or more polyribonucleotides provided herein. In some embodiments, a composition comprises one or more RNA constructs provided herein.
a composition comprises (i) one or more polyribonucleotides and (II) lipid nanopartides, polyptexes (PLX), lipidated polyptexes (LPLX), or liposomes.
the one or more polyribonudeotldes are fully or partially encapsulated within the lipid nanopartides, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.
a composition comprises (i) one or more polyribonudeotldes and (li) lipid nanopartides.
the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticles.
a pharmaceutical composition comprises (I) one or more polyribonucleotides as described herein, one or more RNA constructs as described herein, or a composition as described herein, and (II) at least one pharmaceutically acceptable exdpient [0219]
a pharmaceutical comprises a cryoprotectant.
a cryoprotectant Is sucrose.
a pharmaceutical comprises an aqueous buffered solution, optionally wherein the aqueous buffered solution comprises one or more of Tris base, Tris HCI, NaCI, KCI, NazHPCU, and KHzPO «.
the present disdosure provides, among other things, a combination comprising (i) a first pharmaceutical composition comprising a first polyribonudeotide, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, and (ii) a second pharmaceutical composition comprising a second polyribonudeotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides selected from one or more CyRPA polypeptides or antigenic portions thereof, one or more Ripr polypeptides or antigenic portions thereof, one or more P113 polypeptides or antigenic portions thereof, one or more TRAMP polypeptides or antigenic portions thereof, one or more CSS polypeptides or antigenic portions thereof, or a combination thereof.
a second polypeptide comprises one or more one or more CyRPA polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more Pl 13 polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more Ripr polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more TRAMP polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more CSS polypeptides or antigenic portions thereof.
the present disclosure also provides a combination comprising (i) a first pharmaceutical composition comprising a first polyribonudeotide, wherein the first polyribonudeotide encodes a first polypeptide, and the first pdypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more PiasmodiumT cell antigens.
the present disclosure provides a combination comprising: (i) a first pharmaceutical composition comprising a first polyribonucleotide encoding a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium CSV polypeptides or antigenic portions thereof.
a method comprising administering a polyribonucleotide as described herein to a subject.
the present disclosure also provides a method comprising administering an RNA construct described herein to a subject.
the present disclosure further provides a method comprising administering a composition described herein to a subject.
a method comprising administering one or more doses of the pharmaceutical composition described herein to a subject.
a pharmaceutical composition as provided herein Is for use in the treatment of a malaria Infection comprising administering one or more doses of the pharmaceutical composition to a subject.
a pharmaceutical composition as provided herein Is for use in the prevention of a malaria Infection comprising administering one or more doses of the pharmaceutical composition to a subject [0228] In some embodiments, administering two or more doses of the pharmaceutical conrposltlon to a subject. In some embodiments, administering three or more doses of the pharmaceutical composition to a subject
a first pharmaceutical composition and the second pharmaceutical composition are administered on the same day. In some embodiments, a first pharmaceutical composition and the second pharmaceutical composition are administered on different days. In some embodiments, a pharmaceutical composition and the second pharmaceutical composition are administered to the subject at (Afferent locations on the subjects body.
a method is a method of preventing a malaria infection.
a subject has or is at risk of developing a malaria infection.
a subject is a human.
administration induces an anti-malaria immune response in the subject.
an anti-malaria immune response in the subject comprises an adaptive immune response.
an anti-malaria immune response in the subject comprises a T-cell response.
a T-cell response is or comprises a CD4+ T cell response.
a T-cell response is or comprises a CD6+ T cell response.
an anti-malaria immune system response comprises a B-cell response.
an anti-malaria immune system response comprises the production of antibodies directed against the one or more Plasmodium antigens.
the present disdosure provides a use of the pharmaceutical composition as described herein in the prevention of a malaria infection.
the present disdosure provides a use of the pharmaceutical composition as described herein in inducing an anti-malaria immune response in a subject.
the present disdosure provides a polypeptide encoded by a polyribonucleotide described herein or an RNA construct described herein.
the present disclosure also provides a host cell comprising a polyribonucleotide described herein or an RNA construct described herein.
the present disclosure further provides a host cell comprises a polypeptide described herein.
FIG. 1 provides an annotated polypeptide corresponding to the P. falciparum Rh5 sequence set out in SEQ ID NO: 1. Certain sequence features are annotated, including secretory signal sequence (red bold corresponds to the signal peptide as described by Baum et al. 2009, the additional 2 red residues are included in the signal peptide description by Wright et al.
glycosylation sites NXS/T, blue and bold, lower prediction scores in blue only, the motif in brackets is proximal to the basigin binding site
Rh5 PMX cleavage site black bold
the 3D7 reference sequence has a cysteine at position 203 (red) which is often a tyrosine in naturaNy circulating parasite isolates, paired cysteines are color coded accordingly, and underlined regions are ordered and form the alpha helical composition of the kite structure of the protein, which is associated with basigin binding.
FIG. 2 includes schematics of exemplary polypeptides encoded by poiyribonudeotkles provided herein. All amino add (aa) references induded refer to the P. falciparum Rh5 sequence set out in SEQ ID NO: 1.
(A) includes a schematic of an exemplary polypeptide, RNA Construct 1, encoded by a polyribonudeotlde described herein.
RNA Construct 1 comprises an KSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42), an Rh5 N-terminal "N" disordered region (aa 25-139), an Rh5 ordered region (aa 140-247), an Rh5 linking "L” disordered region (aa 248-296), and an Rh5 ordered region (aa 297-526).
GD KSV glycoprotein D
B indudes a schematic of an exemplary polypeptide, RNA Construct 2, encoded by a polyribonucleotide described herein.
RNA Construct 2 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, an Rh5 ordered region (aa 297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and a foldon domain.
C indudes a schematic of an exemplary polypeptide, RNA Construct 3, encoded by a polyribonudeotlde described herein.
RNA Construct 3 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, and an Rh5 ordered region (297-526).
HSV gD secretory signal e.g., SEQ ID NO: 42
Rh5 ordered region aa 140-247
Rh5 ordered region 297-526.
D indudes a schematic of an exemplary polypeptide, RNA Construct 4, encoded by a polyribonucleotide described herein.
RNA Construct 4 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, an Rh5 ordered region (297-526), a glydne-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain.
HSV gD secretory signal e.g., SEQ ID NO: 42
Rh5 ordered region a 140-247
a glydne-serine linker e.g., SEQ ID NO: 137
ferritin domain e.g., ferritin domain
Membrane RNA Construct 5 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, an Rh5 ordered region (297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
an Rh5 ordered region a 140-247
a glydne-serine linker e.g., SEQ ID NO: 86
HSV gD transmembrane region e.g., SEQ ID NO: 75.
F indudes a schematic of an exemplary polypeptide, RNA Construct 6, encoded by a polyribonucleotide described herein.
RNA Construct 6 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526) comprising an N297Q mutation, a glycine-serine linker (e.g., SEQ ID NO: 86), and a foldon domain.
HSV gD secretory signal e.g., SEQ ID NO: 42
an Rh5 ordered region a 140-247 comprising a C203Y mutation and an N214Q mutation
an Rh5 ordered region aa 297-526
a glycine-serine linker e.g., SEQ ID NO: 86
G indudes a schematic of an exemplary polypeptide, RNA Construct 7, encoded by a polyribonucleotide described herein.
RNA Construct 7 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526) comprising an N297Q mutation.
H indudes a schematic of an exemplary polypeptide, RNA Construct 8, encoded by a polyribonucleotide described herein.
Self assembling RNA Construct 8 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297- 526) comprising an N297Q mutation, a glydne-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain.
HSV gD secretory signal e.g., SEQ ID NO: 42
an Rh5 ordered region a 140-247 comprising a C203Y mutation and an N214Q mutation
an Rh5 ordered region aa 297- 5236 comprising an N297Q mutation
a glydne-serine linker e.g., SEQ ID NO: 137
ferritin domain e.g., SEQ ID NO: 137
RNA Construct 9 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526) comprising an N297Q mutation, a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75).
J indudes a schematic of an exemplary polypeptide, RNA Construct 10, encoded by a polyribonucleotide described herein.
RNA Construct 10 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and a foldon domain.
HSV gD secretory signal e.g., SEQ ID NO: 42
an Rh5 ordered region a 140-247 comprising a C203Y mutation and an N214Q mutation
an Rh5 ordered region aa 297-526
a glydne-serine linker e.g., SEQ ID NO: 86
RNA Construct 11 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526).
HSV gD secretory signal e.g., SEQ ID NO: 42
Rh5 ordered region aa 140-247
aa 297-526 an Rh5 ordered region
RNA Construct 12 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526), a glycine-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain.
HSV gD secretory signal e.g., SEQ ID NO: 42
an Rh5 ordered region a 140-247 comprising a C203Y mutation and an N214Q mutation
an Rh5 ordered region aa 297-526
a glycine-serine linker e.g., SEQ ID NO: 137
ferritin domain e.g., SEQ ID NO: 137
RNA Construct 13 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
an Rh5 ordered region a 140-247 comprising a C203Y mutation and an N214Q mutation
an Rh5 ordered region aa 297-526
a glydne-serine linker e.g., SEQ ID NO: 86
HSV gD transmembrane region e.g., SEQ ID NO: 75
FIG. 3 provides an annotated polypeptide corresponding to the P. falciparum CyRPA sequence set out In SEQ ID NO: 3. Certain sequence features are annotated, Indudlng secretory signal sequence (red) and glycosylation sites (NXS/T, blue).
FIG. 4, parts (A) through (F), includes schematics of exemplary polypeptide encoded by polyribonudeotides provided herein. All amino add (aa) references induded refer to the P. falciparum C/RPk sequence set out in SEQ ID NO: 3.
(A) includes a schematic of an exemplary polypeptide, RNA Construct 14, encoded by a polyribonudeotkie described herein.
RNA Construct 14 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362), a glydne-serine linker (e.g., SEQ ID NO: 86), and a foldon tag.
RNA Construct 15 encoded by a polyribonucleotide described herein.
C includes a schematic of an exemplary polypeptide, RNA Construct 16, encoded by a polyribonucleotide described herein.
RNA Construct 16 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362), a glydne-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
CyRPA a 30-362
a glydne-serine linker e.g., SEQ ID NO: 86
HSV gD transmembrane region e.g., SEQ ID NO: 75.
D includes a schematic of an exemplary polypeptide, RNA Construct 17, encoded by a polyribonucleotide described herein.
RNA Construct 17 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino acid positions 145, 332, and 338, as numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 86), and a foidon tag.
E includes a schematic of an exemplary polypeptide, RNA Construct 18, encoded by a polyribonucleotide described herein.
RNA Construct 18 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), and a portion of CyRPA (aa 30-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at three glycosylation sites.
F includes a schematic of an exemplary polypeptide, RNA Construct 19, encoded by a polyribonudeotlde described herein.
RNA Construct 19 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites, a glycine-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
CyRPA a 30-362
N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites
a glycine-serine linker e.g., SEQ ID NO: 86
HSV gD transmembrane region e.g., S
FIG. 5 Includes schematics of exemplary polypeptide encoded by polyribonucleotides provided herein. All amino add (aa) references Included refer to the P. fatoparum CyRPA sequence set out h SEQ ID NO: 3. Polypeptides may optionally Include one or more linkers between various regions and/or domains.
Indudes a schematic of an exemplary polypeptide, RNA Construct 22, encoded by a polyribonudeotlde described herein. RNA Construct 22 comprises CyRPA secretory signal (SEQ ID NO: 187) and a portion of CyRPA (aa 29-362).
RNA Construct 23 comprises a CyRPA secretory signal (SEQ 10 NO: 187) and a portion of CyRPA (aa 29-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3).
C indudes a schematic of an exemplary polypeptide, RNA Construct 24, encoded by a polyribonudeotlde described herein.
RNA Construct 24 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42) and a portion of CyRPA (aa 29- 362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3).
D includes a schematic of an exemplary polypeptide, RNA Construct 25, encoded by a polyribonucleotide described herein.
RNA Construct 25 comprises a full-length CyRPA (aa 1-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
E Indudes a schematic of an exemplary polypeptide, RNA Construct 26, encoded by a polyribonucleotide described herein.
RNA Construct 26 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CyRPA (aa 29-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
(F) includes a schematic of an exemplary polypeptide, RNA Construct 51, encoded by a polyribonucleotide described herein.
RNA Construct 51 comprises a full-length CyRPA (aa 1-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
G indudes a schematic of an exemplary polypeptide, RNA Construct 52, encoded by a polyribonudeotlde described herein.
RNA Construct 52 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CyRPA (aa 29- 362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO:42
CyRPA a 29- 362
N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332,
FIG. 6, parts (A) and (8) includes schematics of exemplary polypeptide encoded by pdyribonudeotides provided herein. All amino add (aa) references included refer to the P. falciparum P113 sequence set out in SEQ ID NO: 6.
(A) includes a schematic of an exemplary polypeptide, RNA Construct 20, encoded by a polyribonudeotlde described herein.
RNA Construct 20 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of P113 (aa 23-939), a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
P113 aa 23-939
a glydne-serine linker e.g., SEQ ID NO: 86
HSV gD transmembrane region e.g., SEQ ID NO: 75.
B includes a schematic of an exemplary polypeptide, RNA Construct 21, encoded by a polyribonudeotlde described herein.
RNA Construct 21 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of P113 (aa 23-939) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at eight glycosylation sites (e.g., at amino add positions 207, 268, 317, 360, 661, 697, 779, 876, 938, as numbered according to SEQ ID NO: 6), a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
P113 a 23-939
N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at eight glycosylation sites (e
FIG. 7 provides an annotated polypeptide corresponding to the A falciparum Ripr sequence set out In SEQ ID NO: 2. Certain sequence features are annotated, including secretory signal sequence (red), glycosylation sites (NXS/T, blue, stronger predictions in bold), Ripr PMX cleavage site (green), epidermal growth factor domains as described by Chen et al. 2011 are underlined, and the domain In bold Is EGF 7 which has been associated with driving the strongest growth Inhibitory responses (Healer 2019, Nagaoka 2020).
FIG. 8 parts (A) through (L) Includes schematics of exemplary polypeptides encoded by pdyribonudeotides provided herein. All amino add (aa) references included refer to the P. falciparum Ripr sequences set out in SEQ ID NO: 2. Polypeptides may optionally Include one or more linkers between various regions and/or domains.
(A) includes a schematic of an exemplary polypeptide, RNA Construct 27, encoded by a polyribonudeotlde described herein.
RNA Construct 27 comprises a Ripr secretory signal (SEQ ID NO: 186) and a portion of Ripr (aa 20- 1086), a glycine-serine linker (e.g., SEQ ID NO: 138).
B includes a schematic of an exemplary polypeptide, RNA Construct 28, encoded by a polyribonudeotlde described herein.
RNA Construct 28 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20-1086) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
RNA Construct 29 comprises a Ripr secretory signal (SEQ ID NO: 186) and a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at twelve glycosylation sites (e.g., at amino add positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2).
SEQ ID NO: 186 Ripr secretory signal
aa 20-1086 portion of Ripr comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at twelve glycosylation sites (e.g., at amino add positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO
RNA Construct 30 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20- 1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino acid positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
SEQ ID NO: 186 Ripr secretory signal
aa 20- 1086 a portion of Ripr comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino acid positions 103, 144, 228, 303, 334
RNA Construct 31 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42) and a portion of Ripr (aa 20-1086).
F indudes a schematic of an exemplary polypeptide, RNA Construct 32, encoded by a polyribonucleotide described herein.
RNA Construct 32 comprises a an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of Ripr (aa 20-1086), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
G includes a schematic of an exemplary polypeptide, RNA Construct 33, encoded by a polyribonucleotide described herein.
RNA Construct 33 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42) and a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino acid positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2).
H includes a schematic of an exemplary polypeptide, RNA Construct 34, encoded by a polyribonucleotide described herein.
RNA Construct 34 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino add positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr a 20-1086
N-X-[T/S] to Q-X-[T/S] mutations where X is not proline
twelve glycosylation sites e.g., at amino add positions 103, 144, 228,
RNA Construct 53 encoded by a polyribonucleotide described herein.
RNA Construct 53 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20-1086), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
SEQ ID NO: 186 Ripr secretory signal
aa 20-1086 a portion of Ripr
a glydne-serine linker e.g., SEQ ID NO: 138
HSV gD transmembrane region e.g., SEQ ID NO: 75.
J indudes a schematic of an exemplary polypeptide, RNA Construct
RNA Construct 54 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20-1086) comprising N-X-(T/S] to Q-X-[T/S] mutations (where X Is not proline) at twelve glycosylation sites (e.g., at amino acid positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ JD NO: 2), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
SEQ ID NO: 186 Ripr secretory signal
aa 20-1086 a portion of Ripr comprising N-X-(T/S] to Q-X-[T/S] mutations (where X Is not proline) at
RNA Construct 55 encoded by a polyribonudeotide described herein.
RNA Construct 55 comprises a an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of Ripr (aa 20-1086), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO:42
Ripr a 20-1086
a glydne-serine linker e.g., SEQ ID NO: 138
HSV gD transmembrane region e.g., SEQ ID NO: 75.
RNA Construct 56 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino add positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr a 20-1086
N-X-[T/S] to Q-X-[T/S] mutations (where X is not
FIG. 9, parts (A) through (L) includes schematics of exemplary polypeptides encoded by pdyribonudeotides provided herein. All amino add (aa) references included refer to the P. falciparum Ripr sequences set out in SEQ ID NO: 2. Polypeptides may optionally indude one or more linkers between various regions and/or domains.
(A) includes a schematic of an exemplary polypeptide, RNA Construct 35, encoded by a polyribonudeotide described herein.
RNA Construct 35 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 560-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino add positions 646, 964, and 1021 as numbered according to SEQ ID NO: 2), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
B indudes a schematic of an exemplary polypeptide, RNA Construct 36, encoded by a polyribonudeotide described herein.
RNA Construct 36 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 720-934), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
C indudes a schematic of an exemplary polypeptide, RNA Construct 37, encoded by a polyribonudeotide described herein.
RNA Construct 37 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769- 900) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
(D) indudes a schematic of an exemplary polypeptide, RNA Construct 38, encoded by a polyribonucleotide described herein.
Membrane Ripr comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769-856) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
(E) indudes a schematic of an exemplary polypeptide, Membrane Ripr, encoded by a polyribonudeotide described herein.
RNA Construct 38 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-900), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
F includes a schematic of an exemplary polypeptide, RNA Construct 40, encoded by a polyribonudeotide described herein.
RNA Construct 40 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-856), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
RNA Construct 57 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 560-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino acid positions 646, 964, and 1021 as numbered according to SEQ ID NO: 2), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr a 560-1086
Ripr a 560-1086
N-X-[T/S] to Q-X-[T/S] mutations where X is not proline
twelve glycosylation sites
RNA Construct 58 includes a schematic of an exemplary polypeptide, RNA Construct 58, encoded by a pdyribonudeotide described herein.
RNA Construct 58 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 720-934), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr aa 720-934
a glydne-serine inker e.g., SEQ ID NO: 138
HSV gD transmembrane region e.g., SEQ ID NO: 75.
I indudes a schematic of an exemplary polypeptide,
RNA Construct 59 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769-900), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr a 769-900
a glydne-serine inker e.g., SEQ ID NO: 138
HSV gD transmembrane region e.g., SEQ ID NO: 75.
Indudes a schematic of an exemplary polypeptide, RNA Construct 60, encoded by a pdyribonudeotide described herein.
RNA Construct 60 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769-856), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr a 769-856
a glydne-serine inker e.g., SEQ ID NO: 138
HSV gD transmembrane region e.g., SEQ ID NO: 75.
RNA Construct 61 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-900), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr a 817-900
a glydne-serine inker e.g., SEQ ID NO: 138
HSV gD transmembrane region e.g., SEQ ID NO: 75.
RNA Construct 62 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-856), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
Ripr aa 817-856
a glydne-serine inker e.g., SEQ ID NO: 138
HSV gD transmembrane region e.g., SEQ ID NO: 75.
FIG. 10 provides an annotated polypeptide corresponding to the P. anparum'VV.NAV sequence set out in SEQ ID NO: 4. Certain sequence features are annotated, induding secretory signal sequence (red), glycosylation sites (NXS/T, blue), TRAMP deavage site (bold), and SUB2 deavage site (green).
RNA Construct 42 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42) and a portion of TRAMP (aa 42-352) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at eight glycosylation sites (e.g., at amino acid positions, 112, 149, 155, 170, 195, 202, 253, and 305, as numbered according to SEQ ID NO: 4).
HSV gD secretory signal e.g., SEQ ID NO:42
TRAMP a portion of TRAMP (aa 42-352) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at eight glycosylation sites (e.g., at amino acid positions, 112, 149, 155, 170, 195, 202, 253, and 305, as numbered according to SEQ ID NO: 4).
RNA Construct 43 includes a schematic of an exemplary polypeptide, RNA Construct 43, encoded by a polyribonucleotide described herein.
RNA Construct 43 comprises an HSV gD secretory signal (SEQ ID NO:42) and a portion of TRAMP (aa 42-352) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino acid positions, 149, 195, and 202, as numbered according to SEQ ID NO: 4).
FIG. 12 provides an annotated polypeptide corresponding to the P. falciparum CSS sequence set out in SEQ ID NO: 5. Certain sequence features are annotated, including secretory signal sequence (red), glycosylation sites (NXS/T, blue), domains DI and D2 are underlined and joined by the linker region in purple.
FIG. 13, parts (A) through (K) includes schematics of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino add (aa) references Included refer to the P. falciparum CSS sequences set out in SEQ ID NO: 5. Polypeptides may optionally indude one or more linkers between various regions and/or domains.
(A) includes a schematic of an exemplary polypeptide, RNA Construct 44, encoded by a polyribonudeotide described herein. RNA Construct 44 comprises a CSS secretory signal (SEQ ID NO: 188) and a portion of CSS (aa 21- 290).
RNA Construct 45 includes a schematic of an exemplary polypeptide, RNA Construct 45, encoded by a polyribonucleotide described herein.
RNA Construct 45 comprises a CSS secretory signal (SEQ ID NO: 188) and a portion of CSS (aa 21- 290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino add positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5).
C includes a schematic of an exemplary polypeptide, RNA Construct 46, encoded by a polyribonudeotide described herein.
RNA Construct 46 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42) and a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5).
SEQ ID NO:42 HSV gD secretory signal
aa 21-290 CSS
N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5).
D Includes a schematic of an exemplary polypeptide, RNA Construct 47, encoded by a polyribonudeo
RNA Construct 47 comprises a CSS secretory signal (SEQ ID NO: 188), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino add positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
E indudes a schematic of an exemplary polypeptide, RNA Construct 48, encoded by a polyribonucleotide described herein.
RNA Construct 48 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q- X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino add positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
F includes a schematic of an exemplary polypeptide, RNA Construct 49, encoded by a polyribonucleotide described herein.
RNA Construct 49 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-152) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at two glycosylation sites (e.g., at amino add positions 74 and 88, as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
G includes a schematic of an exemplary polypeptide, RNA Construct 50, encoded by a polyribonudeotide described herein.
RNA Construct 50 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 153-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at four glycosylation sites (e.g., at amino acid positions 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
H indudes a schematic of an exemplary polypeptide, RNA Construct 63, encoded by a polyribonudeotide described herein.
RNA Construct 63 comprises a CSS secretory signal (SEQ ID NO: 188), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
SEQ ID NO: 188 CSS secretory signal
aa 21-290 comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283
RNA Construct 64 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO:42
a portion of CSS a 21-290
CSS a 21-290
RNA Construct 65 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-152) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at two glycosylation sites (e.g., at amino add positions 74 and 88, as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO:42
a portion of CSS a 21-152
N-X-[T/S] to Q-X-[T/S] mutations where X Is not proline
a glydne-serine linker e.g., SEQ ID
RNA Construct 66 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 153-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at four glycosylation sites (e.g., at amino add positions 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
FIG. 14, parts (A) through (F) depicts In-vitro host cell viabllty, transfection rate, and expression of polyribonudeotide constructs encoding Plasmodium polypeptides as described herein by host cells (e.g., HEK293T cells).
host cells e.g., HEK293T cells.
A depicts the percentage of viable host cells that are positive for presence of Intracellularly expressed protein for both transfected and non-transfected (NT) cells.
NT non-transfected
B percentage of viable host cells that are positive for presence of surface expressed protein for both transfected and non-transfected cells.
C depicts transfection rate of Indicated polyribonudeotide constructs as measured by percentage of total host cell population that are positive for presence of Intracellularly expressed protein.
(D) depicts transfection rate of Indicated polyribonudeotide constructs as measured by percentage of total host cell population that are positive for presence of surface expressed protein.
(E) depicts total Intracellular protein expression as measured by median fluorescence Intensity of the total host cell population for both transfected and non-transfected cells.
(F) depicts total surface protein expression as measured by median fluorescence of the total host cell population for both transfected and non-transfected cells. Numbers depicted at x-axis indicate RNA construct number. NT stands for non-transfected.
A depicts transfection rate of indicated polyribonucleotide constructs as measured by percentage of total host cell population that are positive for presence of intracellularly expressed protein.
B depicts transfection rate of indicated polyribonucleotide constructs as measured by percentage of total host cell population that are positive for presence of surface expressed protein.
C depicts total intracellular protein expression as measured by median fluorescence intensity of the total host cell population for both transfected and non-transfected cells.
D depicts total surface protein expression as measured by median fluorescence of the total host cell population for both transfected and non- transfected cells. Numbers depicted at x-axis indicate RNA construct number. NT stands for non-transfected.
Construct 1 which encodes RH5, was induded as a control, as ELISA and IVE data with monodonal antibodies have already confirmed immunogenidty for these constructs.
Construct 1 was stained with both the monoclonal antibody 9AD4 at 1:2000 dilution and serum from mice immunized with construct 1.
(B) depicts total protein expression as measured by median fluorescence intensity of the total host cell population for both non-transfected and transfected cells. For all constructs, un-transfected cells (-) were induded as a negative control, (+) indicates cells that were transfected with the matching construct before staining. Numbers depicted at x-axIs indicate RNA construct number. NT stands for non-transfected.
(C) depicts total cell-assodated protein expression determined with the HiBit luminescence assay for CyRPA constructs and (D) depicts total exported protein detected in the culture medium for CyRPA constructs.
(E) depicts total cell-assodated protein expression determined with the HiBit luminescence assay for Rlpr constructs, and (F) depicts total exported protein detected In the culture medium for Rlpr constructs.
G) depicts total cell- assodated protein expression determined with the HiBit luminescence assay for CSS constructs, and (H) depicts total exported protein detected In the culture medium for CSS constructs.
(I) depicts total cell-assodated protein expression determined with the HiBit luminescence assay fbr TRAMP constructs
(J) depicts total exported protein detected in the culture medium fbr TRAMP constructs. Numbers depicted at x-axis Indicate RNA construct number.
FIG. 18 depicts ELISA endpoint reciprocal titers for day 35 serum samples of mice Immunized twice with lug of RH5 encoding constructs. Negative control samples are from mice immunized with a construct encoding CSP. Numbers depicted at x-axis indicate RNA construct number.
FIG. 19, parts (A) through (D), Includes schematics of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino add (aa) references included refer to the P. falciparum GW? sequence set out in SEQ ID NO: 23.
(A) indudes a schematic of an exemplary polypeptide, RNA Construct 77, encoded by a polyribonucleotide described herein.
RNA Construct 77 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a GARP-A region (aa 410-673) (SEQ ID NO: 312).
RNA Construct 78 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), and a GARP-A region (aa 410-673) (SEQ ID NO: 312) comprising a N504Q mutation (SEQ ID NO: 313).
C indudes a schematic of an exemplary polypeptide, RNA Construct 82, encoded by a polyribonucleotide described herein.
RNA Construct 82 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a GARP-A region (aa 410-673) (SEQ ID NO: 312) comprising a N504Q mutation (SEQ ID NO: 312), a glydne-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
D indudes a schematic of an exemplary polypeptide, RNA Construct 84, encoded by a polyribonudeotlde described herein.
RNA Construct 84 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a GARP-A region (aa 52-673) (SEQ ID NO: 342) comprising N183Q, N233Q, N242Q, and N504Q mutations (SEQ ID NO: 309), and a glycine-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).
HSV gD secretory signal e.g., SEQ ID NO: 42
a GARP-A region a 52-673
SEQ ID NO: 342 comprising N183Q, N233Q, N242Q, and N504Q mutations
FIG. 20, parts (A) through (C), includes schematics of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino add (aa) references included refer to the P. faldpanim"VMNP (e.g., PTRAMP) sequence set out in SEQ ID NO: 4.
(A) includes a schematic of an exemplary polypeptide, RNA Construct 133, encoded by a polyribonudeotide described herein.
RNA Construct 133 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a TRAMP region (aa 42-352).
gD HSV glycoprotein D
SEQ ID NO: 42 secretory signal
TRAMP region a 42-352
RNA Construct 134 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a TRAMP region (aa 42-352) comprising N112Q, N129Q, N155Q, N170Q, N195Q, N202Q, N253Q, and N305Q mutations.
gD HSV glycoprotein D
aa 42-352 TRAMP region
RNA Construct 135 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a TRAMP region (aa 42-352) comprising N149Q, N195Q, and N202Q mutations.
gD HSV glycoprotein D
SEQ ID NO: 42 secretory signal
TRAMP region aa 42-352
structures depicted herein are meant to include all stereoisomeric (e.g., enantiomeric or diastereomeric) forms of the structure, as well as all geometric or conformational isomeric forms of the structure.
the R and S configurations of each stereocenter are contemplated as part of the disclosure. Therefore, single stereochemical Isomers, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of provided compounds are within the scope of the disclosure.
provided compounds show one or more stereoisomers of a compound, and unless otherwise indicated, represents each stereoisomer alone and/or as a mixture.
all tautomeric forms of provided compounds are within the scope of the disclosure.
structures depicted herein are meant to Include compounds that differ only In the presence of one or more Isotoplcalty enriched atoms.
compounds having the present structures including replacement of hydrogen by deuterium or tritium, or replacement of a carbon by 13C- or 14C-enrkhed carbon are within the scope of this disclosure.
an agent may be characterized by a particular feature and/or effect.
the term "therapeutic agent' refers to a physical entity has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect
an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic add, saccharide, lipid, metal, or a combination or complex thereof.
amino acid refers to a compound and/or substance that can be, is, or has been incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds.
an amino acid has the general structure H2N-C(H)(R)-C00H.
an amino add is a naturally-occurring amino acid.
an amino acid is a nonnatural amino acid; in some embodiments, an amino add is a D-amlno add; In some embodiments, an amino acid Is an L-amlno add.
Standard amino acid* refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides.
Nonstandard amino add* refers to any amino add, other than the standard amino acids, regardless of whether it is prepared synthetical or obtained from a natural source.
an amino acid, including a carboxy- and/or amino-terminal amino add in a polypeptide can contain a structural modification as compared with the general structure above.
an amino add may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution (e.g., of the amino group, the carboxylic add group, one or more protons, and/or the hydroxyl group) as compared with the general structure.
such modification may, for example, alter the circidating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino add.
such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino add, as compared with one containing an otherwise Identical unmodified amino add.
the term "amino acid' may be used to refer to a free amino add; in some embodiments it may be used to refer to an amino add residue of a polypeptide.
Antigen refers to an agent that (I) elicits an Immune response; and/or (ii) an agent that binds to a T cell receptor (e.g v when presented by an MHC molecule) or to an antibody.
Anti-malaria immuno response refers to an Immune response directed to one or more antigens derived from Plasmodium.
Two events or entities are "associated" with one another, as that term is used herein, if the presence, level, degree, type and/or form of one is correlated with that of the other.
a particular entity e.g., polypeptide, genetic signature, metabolite, microbe, etc.
a particular disease, disorder, or condition if its presence, level and/or form correlates with incidence of, susceptibility to, severity of, stage of, etc. the disease, disorder, or condition (e.g., across a relevant population).
two or more entities are physically "assodated” with one another if they interact, directly or indirectly, so that they are and/or remain in physical proximity with one another.
two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non- covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.
Characteristic portion refers to a portion of a polypeptide or region thereof whose presence (or absence) correlates with presence (or absence) of a particular feature, attribute, or activity of the polypeptide or region thereof.
a characteristic portion of a polypeptide or region thereof is a portion that is found in the polypeptide or region thereof and in related polypeptide or region thereof that share the particular feature, attribute or activity, but not in those that do not share the particular feature, attribute or activity.
a characteristic portion shares at least one functional characteristic with the intact polypeptide or region thereof.
a "characteristic portion" of a polypeptide or region thereof is one that contains a continuous stretch of amino acids, or a collection of continuous stretches of amino acids, that together are characteristic of the polypeptide or region thereof.
each such continuous stretch generally contains at least 2, 5, 10, 15, 20, 50, or more amino acids.
a characteristic portion of a polypeptide or region thereof is one that, In addtion to the sequence and/or structural Identity specified above, shares at least one functional characteristic with the relevant intact polypeptide or region thereof.
a characteristic portion may be biologically active.
a fragment as described herein can be a portion. Accordingly, in some embodiments, a characteristic fragment can be a "characteristic portion.”
Combination therapy refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents (e.g., two or more antibody agents)).
the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all "doses" of a first regimen are administered prior to administration of any doses of a second regimen); In some embodiments, such agents are administered in overlapping dosing regimens.
administration of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modallty(les) in the combination.
combination therapy does not require that Individual agents be administered together In a single composition (or even necessarily at the same time), although In some embodiments, two or more agents, or active moieties thereof, may be administered together In a combination composition.
Comparable refers to two or more agents, entitles, situations, sets of conditions, etc., that may not be Identical to one another but that are sufficiently slmiar to permit comparison there between so that one skilled In the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed.
comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features.
the term “corresponding to” refers to a relationship between two or more entities.
the term “corresponding to' may be used to designate the position/identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition).
a monomeric residue in a pdymer e.g., an amino add residue in a polypeptide or a nucleic acid residue in a polynucleotide
a residue in an appropriate reference polymer may be identified as "corresponding to" a residue in an appropriate reference polymer.
residues in a polypeptide are often designated using a canonical numbering system based on a reference related polypeptide, so that an amino add "corresponding to” a residue at position 190, for example, need not actually be the 190 th amino add in a particular amino add chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify "corresponding" amino acids.
sequence alignment strategies including software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasai, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE that can be utilized, for example, to identify "corresponding" residues In polypeptides and/or nudeic acids in accordance with the present disdosure.
software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasai, PSI-BLAST, PSI-Search
corresponding to may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity).
a gene or protein in one organism may be described as "corresponding to” a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous function and/or that it shows a particular degree of sequence Identity or homology, or shares a particular characteristic sequence element [0274] Dosing regimen-.
dosing regimen may be used to refer to a set of unit doses (typically more than one) that are administered Individually to a subject, typically separated by periods of time.
a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.
Encodes, ks used herein, the term “encode” or “encoding” refers to sequence information of a first molecule that guides production of a second molecule having a defined sequence of nucleotides (e.g., a polyribonucleotide) or a defined sequence of amino adds.
a DMA molecule can encode an RNA molecule (e.g., by a transcription process that Indudes a DNA-dependent RNA polymerase enzyme).
An RNA molecule can encode a polypeptide (e.g., by a translation process).
a gene, a cDNA, or an RNA molecule encodes a polypeptide if transcription and translation of RNA corresponding to that gene produces the polypeptide In a cell or other biological system.
a coding region of a polyribonudeotide encoding a target antigen refers to a coding strand, the nudeotide sequence of which is identical to the polyribonucleotide sequence of such a target antigen.
a coding region of a polyribonudeotide encoding a target antigen refers to a non-coding strand of such a target antigen, which may be used as a template for transcription of a gene or cDNA.
a gene product can be a transcript, e.g., a polyribonucleotide as provided herein.
a gene product can be a polypeptide.
expression of a nudeic add sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splidng, editing, etc.); (3) translation of an RNA into a polypeptide or protein; and/or (4) post-translational modification of a polypeptide or protein.
heterologous As used herein, the term "heterologous", with respect to secretory signal or transmembrane region, refers to a secretory signal or transmembrane region from a virus or an organism other than Piasmocfium.
homolog refers to the overafl relatedness between polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules.
polynucleotide molecules e.g., DNA molecules and/or RNA mdecides
polypeptide molecules are considered to be “homologous” to one another if their sequences are at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical.
polynucleotide molecules e.g., DNA molecules and/or RNA molecules
polypeptide molecules are considered to be "homologous” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., containing residues with related chemical properties at corresponding positions).
certain amino acids are typically classified as similar to one another as “hydrophobic” or “hydrophilic” amino acids, and/or as having "polar' or "non-polar” side chains. Substitution of one amino add for another of the same type may often be considered a "homologous” substitution.
Identity refers to the overall relatedness between polynucleotide molecules DNA molecules and/or RNA molecules) and/or between polypeptide molecules.
polynucleotide molecules eg., DNA molecules and/or RNA molecules
polypeptide molecules are considered to be “substantially Identical” to one another If their sequences are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% Identical.
Calcination of the percent Identity of two nudeic add or polypeptide sequences can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-ldentlcal sequences can be disregarded for comparison purposes).
the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence.
the nucleotides at corresponding positions are then compared.
the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences.
the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent Identity between two nudeotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0).
nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
the percent identity between two nudeotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.
these terms or grammatically comparable comparative terms indicate values that are relative to a comparable reference measurement.
an assessed value achieved with a provided composition e.g., a pharmaceutical composition
an assessed value achieved in a subject may be "increased" relative to that obtained in the same subject under different conditions (e.g., prior to or after an event; or presence or absence of an event such as administration of a composition (e.g., a pharmaceutical composition) as described herein, or in a different, comparable subject (e.g., in a comparable subject that differs from the subject of interest in prior exposure to a condition, e.g., absence of administration of a composition (e.g., a pharmaceutical composition) as described herein.).
comparative terms refer to statistically relevant differences (e.g., that are of a prevalence and/or magnitude sufficient to achieve statistical relevance).
the term “reduced” or equivalent terms refers to a reduction in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or higher, as compared to a comparable reference. In some embodiments, the term “reduced” or equivalent terms refers to a complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero.
the term "increased” or “Induced” refers to an increase In the level of an assessed value by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or higher, as compared to a comparable reference.
In order refers to the order of features from 5' to 3' along the polynucleotide or polyribonucleotide.
in order refers to the order of features moving from the N-termlnal-most of the features to the C- termlnal-most of the features along the polypeptide. "In order” does not mean that no additional features can be present among the listed features.
Isolated means altered or removed from the natural state.
a nucleic add or a peptide naturally present in a living animal is not “Isolated,” but the same nucleic add or peptide partiaHy or completely separated from the coexisting materials of Its natural state is "Isolated.”
An Isolated nucleic add or protein can exist in substantiaRy purified form, or can exist in a non-native environment such as, for example, a host cell.
Linker refers to a portion of a polypeptide that connects different regions, portions, or antigens to one another.
Lipid* As used herein, the terms “lipid” and “lipid-lke material” are broadly defined as molecules which comprise one or more hydrophobic moieties or groups and optionally also one or more hydrophilic moleties or groups. Molecules comprising hydrophobic moieties and hydrophilic moieties are also typically denoted as amphiphiles.
Merozoite stage spedfic Ptasmodium antigen As used herein, the term "merozoite stage specific Plasmodium antigen" refers to an antigen that is expressed during the merozoite stage of the Plasmodium life cyde. In some embodiments, a merozoite stage spedfic Plasmodium antigen is a Rh5 invasion complex antigen.
Multimerization region* refers to a region that directs assembly of multimers into a complex, where each multimer comprises a polypeptide associated with the multimerization region.
RNA lipid nanopartider refers to a nanopartide comprising at least one lipid and RNA molecule(s), e.g., one or more polyribonucleotides as provided herein. In some embodiments, an RNA lipid nanopartide comprises at least one cationic amino lipid.
an RNA lipid nanopartide comprises at least one cationic amino lipid, at least one helper lipid, and at least one polymer- conjugated lipid (e.g., PEG-conjugated lipid).
RNA lipid nanoparticles as described herein can have an average size (.e.g., Z-average) of about 100 nm to 1000 nm, or about 200 nm to 900 nm, or about 200 nm to 800 nm, or about 250 nm to about 700 nm.
RNA lipid nanopartides can have a particle size (e.g., Z-average) of about 30 nm to about 200 nm, or about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, or abort 70 nm to about 80 nm.
an average size of lipid nanoparticles is determined by measuring the average puberte diameter.
RNA lipid nanopartides may be prepared by mixing lipids with RNA molecules described herein.
Neutralization refers to an event in which binding agents such as antibodies bind to a biological active site of a parasite such as a receptor binding protein, thereby inhibiting the parasitic Infedion of cells. In some embodiments, the term “neutralization” refers to an event in which binding agents eliminate or significantly reduce ability of Infecting cells.
nucleic add refers to a polymer of at least 10 nucleotides or more.
a nudek add is or comprises DNA.
a nucleic add is or comprises RNA.
a nucleic add Is or comprises peptide nudek add (PNA).
PNA peptide nudek add
a nudek acid is or comprises a single stranded nudek add.
a nucleic acid is or comprises a double-stranded nucleic add.
a nudek acid comprises both single and doublestranded portions.
a nudek add comprises a backbone that comprises one or more phosphodiester linkages. In some embodiments, a nudek add comprises a backbone that comprises both phosphodiester and non-phosphodlester linkages. For example, in some embodiments, a nudek add may comprise a backbone that comprises one or more phosphorothtoate or S'-N-phosphoramldlte linkages and/or one or more peptide bonds, e.g., as in a "peptide nudek add".
a nudek add comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uradl). In some embodiments, a nudek add comprises on or more, or all, non-natural residues.
natural residues e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uradl.
a non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2- thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5-methykytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridlne, C5-fluorouridfrie, C5-lodouridlne, C5-propynyl-uridlne, C5 - propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8- oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated
a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in natural residues.
a nudek add has a nucleotide sequence that encodes a functional gene product such as an RNA or polypeptide.
a nudek acid has a nucleotide sequence that comprises one or more introns.
a nucleic add may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro), reproduction in a recombinant cell or system, or chemkal synthesis.
a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides long.
a desired reaction In some embodiments relates to Inhibition of the course of the disease (e.g., malaria). In some embodiments, such inhibition may comprise slowing down the progress of a disease (e.g., malaria) and/or interrupting or reversing the progress of the disease (e.g., malaria). In some embodiments, a desired reaction in a treatment of a disease (e.g., malaria) may be or comprise delay or prevention of the onset of a disease (e.g., malaria) or a condition (e.g., a malaria associated condition).
compositions e.g., a pharmaceutical composition
an effective amount of a composition will depend, for example, on disease (e.g., medaria) or a condition (e.g., a malaria associated condition) to be treated, the severity of such a disease (e.g., malaria) or a condition (e.g., a malaria associated condition), Individual parameters of the patient, Including, e.g., age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors.
doses of a composition may depend on various of such parameters.
a reaction In a patient is Insufficient with an initial dose, higher doses (or effectively higher doses achieved by a different, more localized route of administration) may be used.
polypeptide refers to a polymeric chain of amino adds.
a polypeptide has an amino acid sequence that occurs In nature.
a polypeptide has an amino add sequence that does not occur In nature.
a polypeptide has an amino acid sequence that is engineered In that It is designed and/or produced through action of the hand of man.
a polypeptide may comprise or consist of natural amino adds, non-natural amino acids, or both.
a polypeptide may comprise or consist of only natural amino adds or only non-natural amino adds.
a polypeptide may comprise D-amino adds, L-amino acids, or both. In some embodiments, a polypeptide may comprise only D-amino adds. In some embodiments, a polypeptide may comprise only L-amino acids. In some embodiments, a polypeptide may indude one or more pendant groups or other modifications, e.g., modifying or attached to one or more amino add side chains, at the polypeptide's N-terminus, at the polypeptide's C-termlnus, or any combination thereof. In some embodiments, such pendant groups or modifications comprise acetylation, amidation, lipidation, methylation, pegylation, etc., including combinations thereof.
a polypeptide may be cydic, and/or may comprise a cydic portion. In some embodiments, a polypeptide is not cyclic and/or does not comprise any cydic portion. In some embodiments, a polypeptide is linear. In some embodiments, a polypeptide may be or comprise a stapled polypeptide. In some embodiments, the term "polypeptide" may be appended to a name of a reference polypeptide, activity, or structure; in such instances it is used herein to refer to polypeptides that share the relevant activity or structure and thus can be considered to be members of the same class or family of polypeptides.
exemplary polypeptides within the dessert whose amino acid sequences and/or functions are known; in some embodiments, such exemplary polypeptides are reference polypeptides for the polypeptide class or family, in some embodiments, a member of a polypeptide class or family shows significant sequence homology or identity with, shares a common sequence motif (e.g., a characteristic sequence element) with, and/or shares a common activity (in some embodiments at a comparable level or within a designated range) with a reference polypeptide of the ensemble; in some embodiments with all polypeptides within the class).
exemplary polypeptides are reference polypeptides for the polypeptide class or family
a member of a polypeptide class or family shows significant sequence homology or identity with, shares a common sequence motif (e.g., a characteristic sequence element) with, and/or shares a common activity (in some embodiments at a comparable level or within a designated range) with a reference polypeptide of the ensemble; in some embodiments with all polypeptid
a member polypeptide shows an overall degree of sequence homology or identity with a reference polypeptide that is at least about 30-40%, and is often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more and/or includes at least one region (e.g., a conserved region that may in some embodiments be or comprise a characteristic sequence element) that shows very high sequence identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99%.
a conserved region that may in some embodiments be or comprise a characteristic sequence element
a conserved region usually encompasses at least 3-4 and often up to 35 or more amino adds; in some embodiments, a conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more contiguous amino adds.
a relevant polypeptide may comprise or consist of a fragment of a parent polypeptide.
a polypeptide is a Plasmodium polypeptide construct described herein.
a Plasmodium polypeptide construct is a polypeptide that indudes one or more malarial proteins, or one or more portions thereof.
a Plasmodium polypeptide construct described herein Indudes at least one region of Plasmodium Rh5 Invasion complex polypeptide or an antigenic a portion thereof.
a Plasmodium polypeptide construct additionally includes one or more additional amino add sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multlmerization region, and/or a linker, as described herein.
Prevent refers to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time. In some embodiments, prevention refers to reducing the risk of developing clinical malaria.
Referencer describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and/or determined substantially simultaneously with the testing or determination of Interest In some embodiments, a reference or control Is a historical reference or control, optionally embodied in a tangible metfium.
a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and/or comparison to a particular possible reference or control.
Rh5 invasion complex- refers to a complex that is present in Plasmodium merozoites that includes Rh5.
a Rh5 invasion complex facilitates merozoite binding to and/or invasion of an erythrocyte.
polypeptides in an Rh5 invasion complex may include one or more of Plasmodium reticulocyte -binding protein homolog 5 (Rh5), Plasmodium Cysteine-Rich Protective Antigen (CyRPA), Plasmodium Rh5-interacting Protein (Ripr), Plasmodium Pl 13, Absmod't/m thrombospondin-related apical merozoite protein (TRAMP), and Plasmodium cysteine-rich small secreted protein (CSS).
Exemplary Rh5 invasion complexes include the RCR complex and the PCRCR complex.
An RCR complex is a trimeric complex comprising Rh5, cysteine-rich protective antigen (CyRPA), and Rh5-interacting protein (Ripr).
a PCRCR complex is a pentameric complex comprising Rh5, cysteine-rich small secreted (CSS), Ripr, CyRPA, and Piasmodum thrombospondin-related apical merozoite protein (PTRAMP).
RibonucMc add or Polyribonucleotide:
ribonucleic add or polyribonudeotide refers to a polymer of ribonucleotides.
an RNA is single stranded.
an RNA Is double stranded.
an RNA comprises both single and double stranded portions.
an RNA can comprise a backbone structure as described In the definition of "Nudeic add / Polynudeotide” above.
RNA can be a regulatory RNA (e.g., siRNA, microRNA, etc), or a messenger RNA (mRNA).
mRNA messenger RNA
an RNA is a mRNA.
a RNA typically comprises at its 3' end a poly(A) region.
an RNA typically comprises at its 5* end an art-recognized cap structure, e.g., for recognizing and attachment of a mRNA to a ribosome to initiate translation.
a RNA is a synthetic RNA.
Synthetic RNAs indude RNAs that are synthesized rh vitro (e.g., by enzymatic synthesis methods and/or by chemical synthesis methods).
a polyribonucleotide encodes a polypeptide, which Is preferably Is a Plasmodium polypeptide construct
Ribonudeotide As used herein, the term ’ribonucleotide” encompasses unmodified ribonudeotldes and modified ribonudeotldes. For example, unmodified ribonudeotldes Indude the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U).
Modified ribonucleotides may include one or more modifications Including, but not limited to, for example, (a) end modifications, e.g., 5' end modifications (e.g., phosphorylation, dephosphorylation, conjugation, Inverted linkages, etc.), 3' end modifications (e.g., conjugation, inverted linkages, etc.), (b) base modifications, e.g. , replacement with modified bases, stabilizing bases, destabilizing bases, or bases that base pair with an expanded repertoire of partners, or conjugated bases, (c) sugar modifications (e.g., at the 2' position or 4' position) or replacement of the sugar, and (d) intemudeoside linkage modifications, Including modification or replacement of the phosphodiester linkages.
end modifications e.g., 5' end modifications (e.g., phosphorylation, dephosphorylation, conjugation, Inverted linkages, etc.), 3' end modifications (e.g., conjugation, inverted linkages, etc.)
ribonucleotide also encompasses ribonucleotide triphosphates including modified and non-modified ribonucleotide triphosphates.
Secretory signal refers to an amino add sequence motif that targets associated polypeptides for translocation to a secretory pathway.
Subject refers to an organism to be administered with a composition described herein, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes.
Typical subjects indude animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans.
a subject is a human subject.
a subject is suffering from a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition).
a subject is susceptible to a disease, disorder, or condition (e.g., malaria and/or a malaria-assodated condition).
a subject displays one or more symptoms or characteristics of a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition). In some embodiments, a subject displays one or more nonspecific symptoms of a disease, disorder, or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., malaria and/or a malaria- assodated condition). In some embodiments, a subject is a patient. In some embodiments, a subject is an individual to whom diagnosis and/or therapy is and/or has been administered.
Susceptible te An individual who is "susceptible to" a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) is one who has a higher risk of developing the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition ) than does a member of the general public.
an individual who is susceptible to a disease, disorder and/or condition e.g., malaria and/or a malaria-associated condition
an individual who is susceptible to a disease, disorder, and/or condition may exhibit symptoms of the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).
an individual who is susceptible to a disease, disorder, and/or condition e.g., malaria and/or a malaria-associated condition
may not exhibit symptoms of the disease, disorder, and/or condition e.g., malaria and/or a malaria-assodated condition.
an Individual who is susceptible to a disease, disorder, and/or condttion e.g., malaria and/or a malaria-assodated condition
will develop the disease, disorder, and/or condition e.g., malaria and/or a malaria- assodated condition.
an Individual who is susceptible to a disease, disorder, and/or condition e.g., malaria and/or a malaria-assodated condition
will not develop the disease, disorder, and/or condition e.g., malaria and/or a malaria-associated condition.
a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition).
a therapeutic agent or therapy is a medical intervention that can be performed to alleviate, relieve, inhibit, present, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.
Transmembrane region refers to a region of a polypeptide that spans a biological membrane, such as the plasma membrane of a cell.
the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce inddence of one or more symptoms or features of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition). Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition).
a disease, disorder, and/or condition e.g., malaria and/or a malaria-assodated condition.
treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition), for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition.
treatment may be administered to a subject at a later-stage of disease, disorder, and/or condition (e.g., malaria and/or a malaria- assodated condition).
variant refers to a molecule that shows significant structural (e.g., primary or secondary) identity with a reference molecule but differs structurally from the reference molecule.
a variant polypeptide or nucleic add may differ from a reference polypeptide or nudeic add as a result of one or more differences in amino acid or nucleotide sequence and/or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nudeic add (e.g v that are attached to the polypeptide or nudeic acid backbone).
Malaria is a mosquito-borne infectious cfisease caused by single-celled eukaryotic Plasmodium parasites that are transmitted by the bite of Anopheles app. mosquitoes (Phillips, M., etai. Malaria. Nat Rev Dis Primers3, 17050, 2017, which is incorporated herein by reference in its entirety).
Mosquitoes that transmit malaria must have been infected through a previous blood meal taken from an infected subject (e.g., a human). When a mosquito bites an infected subject a small amount of blood is taken in containing Malaria parasites.
RT5,S/AS01 Is an adjuvanted protein subunit vaccine that consists of a portion of the major repeat region and the C- terminus of CSP from Plasmodium falciparum fused to the Hepatitis B surface antigen (HBsAg).
the vaccine is a mix of this ACSP-HBsAg compound with HBsAg that forms virus-like particles (RTS,S/AS01; MosquirixTM).
RTS,S is administered according to a regimen that requires four doses: an initial 3-dose schediie given at least 1 month apart, and a 4th dose 15-18 months after dose 3 (see, for example, Vandoolaeghe & Schuerman Expert Rev Vaccines. 15:1481, 2016; PATH_MVI_RTSS_Fact Sheet_042019, each of which Is incorporated herein by reference In its entirety). Reports indicate that RTS,S protects approximately 30% to 50% of children from clinical disease over 18 months. RTS,S has been reported to induce protective antibody and CD4+ T-cell responses, but only negligible CD8+ T cell responses (see, for example, Moris et al.
infected mosquitos inject, along with their anticoagulating saliva, sporozoites known as the liver stage of Plasmodium spp. Sporozoites journey through the skin, and if successful, into hepatocytes of the liver. This journey happens very quickly; it can be completed within only a few minutes (Sinnis et al., Parasite! Int. 2007 Sep;56(3): 171-8, which is incorporated herein by reference in its entirety).
sporozoites When moving from an inoculation site in the skin to the liver, sporozoites traverse host ceHs (Mota et al., Science 2001 Jan 5;291(5501): 141-4). Sporozoites traverse different types of host cells at the dermis, including fibroblasts and phagocytes (Amino et al.. Cell Host Microbe. 2008 Feb 14;3(2):88-96, which is incorporated herein by reference in its entirety), and the liver sinusoidal barrier, containing liver endothelial cells and Kupffer cells (Frevert et al., PLoS Biol 3(6): el92.
yoetiispotozoktes can enter hepatocytes via a transient vacuole and that host membrane rupture occurs upon cell exit rather than cell entry (Risco-Castillo et al., Cell Host Microbe 2015 Nov ll;18(5):593-603, which is incorporated herein by reference in its entirety).
GPDH glyceraldehyde 3- phosphate dehydrogenase
sporozoites Once sporozoites have Invaded liver cells, they dfferentiate into merozoites, a replicative form of the parasite. Within a few days, a single sporozoite can lead to 5,000-10,000 merozoites. Merozoites bud from the host hepatocyte in structures called merosomes, which contain up to a thousand merozoites and are hidden from host immune responses due to their host membrane composition. These merosomes ultimately release merozoites that mix freely with the host cytoplasm and appear in the blood stream, where they invade red blood cells and begin the blood stage of infection, characterized by fast asexual replication and clinical presentation. Within a small number of days, hundreds of thousands of parasites can be present in the blood.
Plasmodium spp. parasites gain entry into red blood cells through specific ligand-receptor interactions mediated by proteins on the surface of the parasite that interact with receptors on the host erythrocyte (mature red blood cell) or reticulocyte (immature red blood cell), whereas P. /atoparumcan invade and replicate in erythrocytes and reticulocytes, P. wVaxand other species predominantly invade reticulocytes, which are less abundant than erythrocytes. Most of the erythrocyte-binding proteins or reticulocyte-binding proteins that have been associated with invasion are redundant or are expressed as a family of variant forms; however, for P. falciparum, two essential red blood cell receptors (basigin and complement decay-accelerating factor (also known as CD55)) have been identified.
basic red blood cell receptors basic and complement decay-accelerating factor (also known as CD55)
P. mar and P. ovale can also enter a dormant state in the liver, the hypnozoite.
the invasion of a red blood cell by the merozoite involves interactions of multiple parasite derived proteins (e.g., ligands) with red blood cell (RBC) proteins (e.g., receptors) (Weiss et al., Pios Path., 2015 Feb 27; 10.1371, which is Incorporated herein by reference in its entirety).
RBC red blood cell
the invasion process begins by the merozoite first attaching to the RBC.
Merozoite-RBC Interaction Is further strengthened through merozoite surface protein- 1 (MSP1) and unknown RBC proteins, causing some deformation of the RBC surface.
MSP1 merozoite surface protein- 1
Rh5 invasion complex comprising Plasmodium thrombospondin-related apical merozoite protein (PTRAMP), cystelne-rlch small secreted (CSS), cysteine-rich protective antigen (CyRPA), Rh5-interactlng protein (Rlpr) and Rh5.
PTRAMP Plasmodium thrombospondin-related apical merozoite protein
SCS cystelne-rlch small secreted
CyRPA cysteine-rich protective antigen
Rh5-interactlng protein Rh5-interactlng protein
Rh5 invasion complex including the five proteins is referred to as the PCRCR complex.
PCRCR complex begins in the endoplasmic reticulum, where Its constituents cystelne-rlch small secreted (CSS) protein and Plasmodium thrombospondin-related apical merozoite protein (PTRAMP) Interact to form a heterodimer referred to as PTRAMP-CSS.
PTRAMP-CSS Is trafficked to a secretory organelle called the microneme, where CSS interacts with Rh5-interactlng protein (Rlpr) and cystelne-rlch protective antigen (CyRPA) to form a tetrameric complex called PCRC.
the merozoite Initiates a polarized secretion process that allows the mlcronemal PCRC complex opportunity to Interact with rhoptry protein Rh5, forming the pentameric complex PCRCR.
Rh5 a host cell receptor
basigin broadly across the merozoite-RBC Interphase.
PCRCR binding to basigin creates a stable and Irreversible platform between the apical end of the merozoite and the deformed RBC surface, allowing Rh5 and Rlpr to embed into the RBC membrane (Scally et al., Nat Microb., 2022 May 4, which is incorporated herein by reference in its entirety).
an open connection between the merozoite's apical tip and the RBC surface acts as a conduit for Ca 2 * to flow into the RBC, and for merozoite derived invasion proteins, such as AMA1 and RON2, to help establish the moving junction (Srinivasan et al., Proc Natl Acad Sd., 2011 Jul 25;13275-80, which is Incorporated herein by reference in its entirety). It Is through this moving junction that the parasite, utilizing its actin-myosin dependent gliding motility, propels itself inside the RBC and establishes a parasitopherous vacuole within which the parasite grows and replicates.
Rh5 binds basigin with higher affinity when complexed with CyRPA and Ripr to form the RCR complex, and has the highest affinity for basigin when part of the PCRCR complex (Wong et al., Nature., 2019 and Scally et al., Nat Microb., 2022 May 4, each of which are incorporated herein by reference in its entirety).
the male and female gametocytes fuse, forming a diploid zygote, which matures and differentiates into an ookinete; this motile form secretes chitinases and other lytic proteins such as CelTOS in order to disrupt and traverse through the peritrophic matrix and traverse the midgut epithelium to reach the basal lamina where it further differentiates and matures as an oocyst Oocysts mature over approximately 10 days (depending on the temperature), replicating to form sporozoites that egress the mature oocyst Into the hemocoel of the mosquito. Thousands of sporozoites form in a single oocyst and become randomly distributed throughout the hemocoel.
sporozoites are passively circulated through the mosquito haemolymph until they encounter the salivary glands, where they actively invade the glands. Following invasion of the salivary gland, sporozoites are re-programmed via an unknown mechanism to prepare for Iver Invasion. Evidence of this reprogramming has been demonstrated by the inability of midgut sporozoites (directly from oocysts) to invade hepatocytes, and also by the fact that sporozoites which have successfully invaded a salivary gland are unable to re-invade another salivary gland if presented one.
Salivary gland sporozoites alter mosquito behavior and salivary gland function, as less saliva is produced resulting In an Increase In mosquito probing behavior, increasing the chances of transmission to a human host via a mosquito bite and continuing the human host's phase of this parasite's life cyde.
P. vivax anti P. ovale can also enter a dormant state in the liver, the hypnozoite.
One particular challenge associated with mounting an immune response capable of clearing sporozoites before a liver infection Is that the most abundant and Immunogenic protein on the sporozoite surface, the circumsporozoite protein (CSP), is only exposed to the immune system in small quantities and for a short duration of time due to the variably low inoculum from the mosquito and the kinetics of hepatocyte infection after inoculation.
CSP circumsporozoite protein
the parasite differentiates into a stage during which it no longer expresses CSP and hstead has a different mosaic of surface antigens, making the parasite Invisible to, e.g., any memory immune response directed towards the liver stage.
the early stages of parasite exposure to the human host create a short window for the innate and memory immune systems to generate a response that is capable of clearing all parasites before infection is established.
Malaria symptoms typically develop 4-8 days after initial red blood cell invasion. Replication cyde of merozoites within the red blood cells continues for 36-72 hours, until hemolysis, releasing the merozoites for another round of red blood cell infection. Thus, in synchronous infections (infections that originate from a single infectious bite), fever occurs every 36-72 hours, when infected red blood cells lyse and release endotoxins en masse. [0325] Some drugs that prevent Plasmodium spp.
the adenine-thymine (AT) content of Plasmodium spp. can also be very different, e.g., ⁇ 80% AT in P. falciparum, P. venezowi, and P. gaHinaceunr, ⁇ 75% AT in rodent malaria parasites; and ⁇ 60% AT in P. vivax, P. knowtesi, and P. cynomolgi. AT content is often higher in introns and intergenic noncoding regions than in proteincoding exons, with an average of 80.6% AT for the whole P. fakiparum genome versus 86.5% for noncoding sequences.
fakiparum reflects large numbers of low-complexity regions, simple sequence repeats, and microsatellites, as well as a highly skewed codon usage bias.
Polymorphisms of AT-rich repeats provide abundant markers for linkage mapping of drug resistance genes and for tracing the evolution and structure of parasite populations.
Malaria parasite genomes carry multigene families that serve important roles in parasite interactions with their hosts, including, for example, antigenic variation, signaling, protein trafficking, and adhesion.
genes encoding P. fakiparum erythrocyte membrane protein 1 (P/EMP1) have been studied most extensively.
P/EMP1 P. fakiparum erythrocyte membrane protein 1
Each Individual P. falciparum parasite carries a unique set of 50 to 150 copies of the vargene In Its genome, where switches of gene expression can produce antigenic variation.
P/EMP1 plays an Important role in the pathogenesis of clinical developments such as in cerebral and placental malaria, in which it mediates the cytoadherence of infected red blood cells (iRBCs; infected erythrocytes) in the deep tissues.
iRBCs infected red blood cells
chabaudi ⁇ df were shown to be expressed in different cellular locations, within and on the suface of iRBCs, and in merozoites. Malaria parasites devote large portions of their genomes to gene families that ensure evasion of host immune defenses and protection of molecular processes essential to infection. These famiies emphasize the importance of research on their roles in parasite-host interactions and virulence, despite the difficulties inherent to their investigation.
An additional, exemplary polymorphic gene family comprises a group of 14 genes encoding proteins with six cysteines (6-Cys). These proteins often localize on the parasite surface interacting with host proteins and are expressed at different parasite developmental stages. 6-Cys proteins also demonstrate diverse functions and have been shown to play roles in, for example, parasite fertilization, mating interactions, evasion of immune responses, and invasion of hepatocytes. The proteins expressed in asexual stages are generally polymorphic and/or under selection, suggesting that they could be targets of the host immune response; however, their functions in parasite development remain largely unknown.
Plasmodium genomes can be highly polymorphic. Early studies demonstrated polymorphisms involving tens to hundreds of kilobases and that the chromosome structure in P. falciparum is largely conserved in central regions but extensively polymorphic is both length and sequence near the telomeres. Much of the subtelomeric variation was explained by recombination within blocks of repetitive sequences and families of genes.
microsatellites The frequency of simple sequence repeats (microsatellites) in P. faldparum is estimated to be approximately one polymorphic microsatellite per kb DNA. Without wishing to be bound by any one theory, this high rate may reflect the AT -rich nature of the genome. Mlcrosatellltes seem to be less frequent In other Plasmodium species that have genomes with lower AT contents.
SNPs Single Nucleotide Polymorphisms
CNVs Copy Number Variations
Rh5 is found In P. faldparum, but not in all other species of Plasmodium that infect humans which contain proteins that perform an analgous function as Rh5.
Rh5 orthologues are also found in other species belonging to the Lavarenia subgenus, which includes parasites that infect chimpanzees and gorillas, indicating a unique role in P. falciparum invasion of human erythrocytes. See, e.g., Ragotte, et al. Trends Parasitol. 36(6) 2020, which is incorporated herein by reference in its entirety.
Rh5 is expressed during the mature schizont stages and localizes to the Rhoptry, a polarized secretory organelle of P. faldparum. Rh5 is a protein that is secreted during the invasion process and is believed to be essential for the completion of parasite invasion of the erythrocyte. Rh5 functions downstream of intial parasite attachment to the erythrocyte surface and deformation of the erythrocyte membrane. Upon secretion, Rh5 binds to erythrocyte surface protein basigin, creating an attachment and a stable platform for downstream receptor-ligand interactions and associated invasion invents to occur.
Bindng of Rh5 to basigin is also required for the induction of a spike in calcium within the erythrocyte, which is blocked when merozoites attempt to invade in the presence of anti-Rh5, anti-Ripr, or anti-baslgin antibodies or soluble basigin (See, e.g., Ragotte (2020), which is incorporated herein by reference in its entirety).
Rh5 can interact with multiple proteins to form complexes.
the affinity of Rh5 for basigin varies based on which complex Rh5 is in.
Rh5 can complex with CyRPA and Ripr to form an elongated protein trimer, called RCR, on the merozoite surface that binds to erythrocyte surface protein basigin with higher affinity than Rh5 alone.
RCR elongated protein trimer
Rh5 has its highest affinity to basigin when interacting with PTRAMP, CSS, Ripr, and CyRPA in a pentameric complex known as PCRCR. See, Scally (2022).
Rh5 is a 63 kDa protein expressed during the mature schizont stage. It is processed and cleaved to a 45 kDa form which is shed by the parasite. The structure of Rh5 reveals a kite-like architecture formed from the coming together of two three-helical bundles. See, e.g., Ragotte (2020), which is incorporated herein by reference in its entirety.
Rh5 sequences are known (see, e.g., UnIProt accession numbers A0A159SK44, A0A159SK99, A0A159SKS8, A0A159SKW8, A0A159SL23, A0A159SL78, A0A159SL96, A0A159SLM7, A0A159SMC8, A0A159SMR9, A0A161FQT0, A0A1B1UZE2, A0A1B1UZE4, A0A1B1UZE5, A0A346RCI1, A0A346RCJ0, A0A346RCJ2, A0A346RCJ3, A0A346RCJ4, A0A346RCK4, A0A346RCK5, A0A346RCK6, A0A346RCK9, B2L3N7, Q8IFM5, each of which is incorporated herein by reference in its entirety), and exemplary Rh5 amino add sequence is provided in Table 1.
Rh5-lnteracting Protein is an approximately 120 kDa protein and localized to mkronemes during the schizont stage of the Plasmodium life cycle. The full-length 120 kDa protein is processed into two fragments of similar size, an N-termlnal fragment (Indudlng EGF domains 1 and 2) and a C-termlnal fragment (Indudlng EGF domains 3-10). Ripr colocalizes with Rh5 and CyRPA during parasite invasion at the junction between merozoites and erythrocyte. Parasites with conditional knockouts of fliRipr induce membrane deformation, but cannot complete Invasion (See, e.g., Ragotte (2020) which is incorporated herein by reference in Its entirety).
Ripr sequences are known (see, e.g., UniProt accession numbers A0A193PDI9, A0A193PDK3, A0A193PDK8, A0A193PDL3, A0A193PDL9, A0A193PDP4, A0A193PDQ8, A0A193PE01, A0A193PE05, A0A193PE07, 097302, A0A193PE17).
Exemplary Ripr amino add sequences are provided In Table 1.
Cysteine-Rich Protective Antigen is a 43 kDa protein with a predicted N-terminal secretion slgial. CyRPA is a critical mediator of an Invasion multi-protein complex consisting of Rh5 and Ripr, known as RCR, that Improves Rh5 affinity to the erythrocytic basigin receptor. The ability of CyRPA to make limited contact with Ripr and Rh5 is thought to permit the disassociation of Rh5 and Ripr from this complex so they can be inserted into the erythrocyte membrane, helping to further stabilize the contact between parasite and erythrocyte. CyRPA also plays a role in the PCRCR complex which further increase Rh5 affinity to basigin.
CyRPA mediated Rh5 binding to basigin is important for stabilizing the parasite-erythrocyte membrane interaction which allows downstream invasion events, like Ca2 + increase In the erythrocyte cytosol and establishment of tight/movlng junctions, to occur. CyRPA is highly conserved, with only a single SNP above 5% prevalence in the general population. CyRPA is important for invasion (as conditional knockdown causes the loss of invasion activity), and CyRPA has poor sero-reactivity from natural exposure (See, e.g., Ragotte (2020) and Cowman (2018) each of which is incorporated herein by reference in its entirety).
CyRPA sequences are known (see, e.g., Uniprot accession number A0A2S1Q7P0, A0A2S1Q7P5, A0A2S1Q7Q4, Q8IFM8, each of which is incorporated herein by reference in its entirety).
Exemplary CyRPA amino add sequence is provided in Table 1.
PTRAMP Ptesmtxi'u/nThrombowondic- Related Apical Merozoite Protein
Plasmodium thrombospondic-related apical merozoite protein is a 352 amino add protein that localizes to developing micronemes and relocates to the merozoite surface upon invasion and is understood to be important for host cell invasion by P. falciparum. Orthologs to the gene encoding PTRAMP is present in all malaria parasite spedes examined, indicating a conserved role in host invasion and making it an opportune target for therapeutic intervention.
PTRAMP appears to exhibit both linear and conformational epitopes, which would give the immune system the opportunity to utilize both B cell receptors and T cell receptors for recognition and activation when exposed to PTRAMP (Alvaro et al., Malaria Jou., 2010 Oct. 13;10.1186, which is Incorporated herein by reference in its entirety).
PTRAMP can interact with cysteine-rich small secreted (CSS) protein to form a heterodimer known as PTRAMP-CSS which in turn can, through CSS, Interact with Ripr and CyRPA to form a tetrameric complex called PCRC, that ultimately Interacts with Rh5 forming a pentameric complex PCRCR which enhances Rh5 binding to host baslgin and Is important for P. falciparum to invade RBCs.
PCRC tetrameric complex
Rh5 tetrameric complex
Nanobodies against PTRAMP inhibit merozoite significantly reduce merozoite invasion and give evidence to the potential efficacy therapeutically targeting PTRAMP could provide (Scally et al., Nat Microb., 2022 May 4, which Is incorporated herein by reference In Its entirety).
PTRAMP sequences are known (see, e.g., Uniprot accession number [Q8I5M8]).
Exemplary PTRAMP amino acid sequence is provided in Table 1.
Cysteine-rich small secreted protein Is a 290 amino add cryptic 6-Cys protein comprised of two degenerate 6-Cys domains. Proteins belonging to this family commonly mediate extracellular protein-protein interactions, a role that has been confirmed to occur In P. falciparum. CSS is capable of biding both PTRAMP and Ripr. CSS Interaction with PTRAMP occurs in the endoplasmic reticulum, whereas CSS interaction with Ripr occurs after trafficking to the micronemes. CSS Interaction with PTRAMP and Ripr are involved in Invasion and the formation of the PCRC complex, which is capable of complexing with Rh5 and enhancing Rh5's Interaction with baslgin.
nanobodies against CSS Similar to PTRAMP, nanobodies against CSS have been showing to Inhibit Invasion, indicating Its role in the invasion process likely thought its partldpation in the PCRCR complex. Nanobodies which inhibit CSS binding to PTRAMP or Ripr do not inhibit invasion, confirming the interaction of these proteins before exposure to the merozoite surface. The Inhibitory activity of nanobodies against CSS are thought to occur by potentially blocking Ripr and/or Rh5 insertion into the RBC membrane and/or disrupting the conformational changes of the PCRCR complex required for invasion, although the exact mechanism of inhibition is not entirely known.
CSS sequences are known (see, e.g., Uniprot accession number [Q8IM47]). Exemplary CSS amino acid sequence is provided in Table 1.
a Plasmodium CSS polypeptide or antigenic portion thereof comprises a serine at position 30, as numbered according to SEQ ID NO: 5.
SEQ ID NO: 5 As exemplary CSS amino add sequence comprising a C30S mutation is provided in Table 1.
P113 is a glycosylphosphatidylinositol (GPI)-linked protein that interacts directly with the N terminus of unprocessed Rh5, which has been thought to provide a mechanism by which the Rh5 invasion complex is tethered to the merozoite surface.
GPI glycosylphosphatidylinositol
P113 is present in the paristopherous vacuole (PV) and has been shown to interact with parasite export complexes like PTEX and EPIC, which are important for the transport of parasite effector proteins across the paristopherous vacuole membrane (PVM).
PVM paristopherous vacuole membrane
P113 orthologues are found in all Plasmodium species sequenced thus far, which is suggestive of common and conserved functions (Bullen et al. (2022) Molecular Microbiology 117:1245-1262, which is incorporated herein by reference in its entirety). Despite this fact, in P.
Plasmodium Pl 13 sequences are known (see, e.g., Uniprot accession number Q8ILP3).
Exemplary P113 amino acid sequence is provided in Table 1.
Plasmodium parasites are known to express various proteins at different stages of their lifecycles. Exemplary malarial proteins are described below, and exemplary amino add sequences are provided in Table 2. Circumsporozoite Protein fCSPI
Circumsporozoite protein is a multifunctional protein that is involved in Plasmodium life cycle, as it is required for the formation of sporozoites in the mosquito midgut, the release of sporozoites from the oocyst, invasion of salivary glands, attachment of sporozoites to hepatocytes in the liver, and sporozoite invasion of hepatocytes (see, e.g., Zhao et al. (2016) PLoS ONE 11(8): e0161607, which is incorporated herein by reference in its entirety).
CSP is present in all Plasmodium spedes, and although variation exists in the amino add sequence across species, the overall domain structure of a central repeat region and nonrepeat flanking regions is well conserved (see, e.g., Zhao et al. (2016) PLoS ONE 11(8): e0161607; Wahl et al. (2022) J. Exp. Med. 219: e20201313, each of which is incorporated herein by reference in its entirety).
CSP sequences are known (see, e.g., UniProt accession numbers A0A2L1CF52, A0A2L,lCF88, C6FGZ3, C6FH2,7 C6FHG7, M1V060, M1V0A3, Ml V0 B0, M1V0C4, M1V0E0, M1V9I4, M1VFN9, M1VKZ2, P02893, Q5EU9, Q5BK2, Q5BK8, Q5EIL3, Q5EIL5, Q5BL8, Q5R2L2, Q7K740, Q8I9G5, Q8I9J3, Q8I9J4).
Exemplary CSP amino acid sequences are provided in Table 2.
E140 is found in every Plasmodium species for which a genomic sequence is available, and is well conserved, with amino add identity ranging from 34-92% among spedes. See, e.g., Smith , et al. PLoS one 15.5 (2020): 60232234; http://doi: 10.1371/joumal.pone.023223; and U.S. Patent Publication No. US 2019/0117752; each of which are incorporated herein by reference in their entirety. E140 is also highly conserved (95-99%) in P. falciparum sir aixxs isolated from different locations around the world, and exhibits a low mutation frequency. E140 is expressed at different life stages of malaria parasites (spedfically, E140 has been detected in sporozoites, liver, and blood stage parasites).
E140 Protein structure algorithms predict that the E140 protein has five transmembrane domains, presumable spanring a parasite or host-derived membrane. E140 displays distinct patterns of protein expression In mature sporozoites, late liver, and late schizont stages. It traffics to the anterior and posterior ends of the sporozoite, the parasitophorous vacuole space of the late liver stage and around developing merozoites in the late schizont stage. It is also known to be expressed In mature salivary gland sporozoites as well as oocyst-derived sporozoites and oocysts.
E140 sequences are known (see, e.g., UnIProt accession numbers A0A650D649, A0A650D653,
CeiTOS Is required for sporozoite traversal through Kupffer cells during the liver invasion process.
CeiTOS forms a pore from within the cell, allowing for sporozoite egress into the liver.
Antibody epitopes have been characterized from Immunized mice and infected human populations (PFand PV). In mouse studies, Immunization with CeiTOS has been shown to provide protection and against challenge.
Vaccination with CeiTOS may generate antibodies that can bind the extracellular domain of the pore-forming complex, blocking complete formation of the pore and preventing sporozoite traversal Into the liver. See, e.g., Jimah et al., ELIFEe2016 Dec l;5:e20621. dol: 10.7554/eUfe.20621, which is incorporated herein by reference in its entirety.
CeiTOS sequences are known (see, e.g., Uniprot accession number M1ETJ8, Q53UB7, A0A2R4QLA5, A0A2R4QU0, A0A2R4QLI5, A0A2R4QU1, A0A2R4QU4, M1ETJ8, Q53UB8, Q8I5P1, each of which is incorporated herein by reference in its entirety).
An exemplary CeiTOS amino add sequence is provided in Table 2.
SPECT1 and SPECT2 are siyiificant Plasmodium proteins that may play a role in cell traversal. See Yang et al., Cell Rep. 2017 Mar 28;18(13):3105-3116. doi: 10.1016/j.celrep.2017.03.017, which is incorporated herein by reference in its entirety.
Targeted disruption of P. falciparum SPECT1 or SPECT2 has been shown to reduce infectivity of sporozoites in liverstage development in humanized mice.
mechanisms of cell traversal of these two proteins are yet to be defined in P. falciparum. See Yang et al.
SPECT1 and SPECT2 are considered attractive pre-erythrocytic immune targets due to the key role they are thought to play in the crossing of the malaria parasite across the dermis and the liver sinusoidal wall, prior to invasion of hepatocytes.
Recombinant P. falciparum SPECT2 has been shown to cause lysis of red blood cells in a Ca 2- * -dependent manner, as has the MACPF/CDC domain of ASPECT2.
ASPECT2 has also been implicated in the Ca2+ -dependent egress of P. falciparum merozoites from red blood cels.
SPECT1 and SPECT? sequences are known (see, e.g., UniProt accession numbers Q8IDR4 and Q9U0J9, each of which is incorporated herein by reference in its entirety), and exemplary amino add sequences for SPECT1 and SPEC!? are provided in Table 2.
Exported protein 1 is a single pass transmembrane protein with an N-terminal signal peptide expressed during intraerythrocytlc stage and liver stage (see, e.g., Spielmann et al., Int J Med Microbiol. 2012 Oct;302(4-5): 179-86, which is Incorporated herein by reference In its entirety).
EXP1 was shown to Initially localize to dense granules in merozoites and then be transported to parasltophorous vacuolar membrane (PVM) after invasion (see, e.g., Iriko et al., Parasitol Int.
PVM parasltophorous vacuolar membrane
EXP1 forms homo-oligomers with a N-terminus that is exposed to the parasltophorous vacuolar lumen and a C-terminus that is exposed to the red blood cell cytosol (see, e.g., Mesén- Ramrez et al., PLoS Biol. 2019 Sep 30;17(9):e3000473, which is Incorporated herein by reference in Its entirety).
EXP1 has been demonstrated to possess glutathione S-transferase (GST) activity that may protect Plasmodium from oxidative damage (see, e.g., Mesén-Ramirez et al., PLoS Biol 17(9) 2019 Sep 30;17(9):e3000473, which is incorporated herein by reference in its entirety). Recently, it was demonstrated that EXP1 is important for Plasmodium survival by maintaining correct localization of EXP2, a nutrient-permeable channel in the PVM (see, e.g., Mesen-Rami'rez et al., PLoS Biol. 2019 Sep 30;17(9):e3000473, which is Incorporated herein by reference in Its entirety).
GST glutathione S-transferase
P. falciparum EXP1 polypeptide sequences are known (see, e.g., UniProt accession number Q8IIF0,
Upregulated in infective sporozoites gene 3 is a membrane-bouxi protein localized to sporozoite parasltophorous vacuolar membrane (PVM) in infected hepatocytes.
UIS3 was shown to interact with liver fatty addbinding protein (L-FABP) and be involved in fatty acid and/or lipid import during phases of Plasmodium growth (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29; 283(35): 24077-24088; Mikolajczak et al., Int J Parasitol. 2007 Apr;37(5):483-9, each of which is incorporated herein by reference in its entirety).
L-FABP liver fatty addbinding protein
Plasmodium structural features e.g., parasltophorous vacuolar membrane.
the Plasmodium relies on host fatty adds for rapid synthesis of its membranes (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29; 283(35): 24077-24088, which is incorporated herein by reference in its entirety).
UIS3 insertion in the PVM provides Plasmodium a method to import essential fatty adds and/or lipids during rapid sporozoites growth phases (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29; 283(35): 24077-24088, which is incorporated herein by reference in its entirety).
UIS3-deficient Plasmodium berghei are also unable to develop into mature liver schizonts and therefore abort malaria infection within the liver itself (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022): 164-7, which is incorporated herein by reference in its entirety). Further, it was previously demonstrated that UIS3 derived from Plasmodium berghei and UIS3 derived from Plasmodium faldparum exhibited a low (i.e. 34%) amino acid sequence identity (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022): 164-7, which is incorporated herein by reference in its entirety).
Plasmodium UIS3 sequences are known (see, e.g., UniProt accession number A0A509ARS3, A0A1C6YLP3, Q8IEU1, A0A384KU1, A0A1G4H423, A0A077YB01, Q9NFU4, each of which is Incorporated herein by reference In Its entirety).
An exemplary U1S3 amino acid sequence Is provided In Table 2.
ETRAMP10.3 Early Transcribed Membrane Protein 10.3 (ETRAMP10.3) and Unregulated in Infective S es Gene 4 (UIS4)
UIS4 Upregulated in infective sporozoites gene 4 (UIS4) contains a single transmembrane domain and localizes to secretory organelles of sporozoites and to the parasitophorous vacuole membrane (PVM) of liver stages. UIS4 is not expressed In blood stages or early sporozoites that are produced in oocysts (see, e.g., Mackellar et al., Eukaryot Cell. 2010 May; 9(5): 784-794, which is incorporated herein by reference in Its entirety).
UIS4 Deletion of UIS4 gene is associated with arrest of early liver stage development (see, e.g., Vaughan and Kappe, Cold Spring Harb Perspect Med. 2017 Jun l;7(6):a025486, which Is Incorporated herein by reference in its entirety). Recently, UIS4 was demonstrated to be Involved in Plasmodium berghei survival by eluding host actin structures deployed as part of host cytosolic defense (see, e.g., Sana et al., (Science. 2022 Apr 22;25(5): 104281. doi: 10.1016/j.isci.2022.104281. eCollection 2022 May 20, which Is Incorporated herein by reference In Its entirety). P.
falciparum has an ortholog to UIS4 named ETRAMP10.3 which is not able serve as a functional compliment to P. yoeliiUIS4, Indicating It likely serves a different function In P. falciparum's lfie cycle (see Mackdlar et al., Eukaryot Cell 9:784-94 (2010), which is incorporated herein by reference In its entirety).
Plasmodium faldparum early transcribed membrane protein 10.3 (ETRAMP10.3) Is an approximately 10 kDa protein and member of the early transcribed membrane proteins multigene family, a family which is conserved across Plasmodium species and includes proteins located In the parasitophorous vacuole.
ETRAMP10.3 is one example, which is expressed in both liver and blood stage P. faldparum parasites. ETRAMP10.3 transcription has been found to peak during the transition from ring to trophozoite stages of P.
ETRAMP10.3 localizes to the parasitophorous vacuole and is exported to a host erythrocyte during blood stage infection.
ETRAMP10.3 is sometimes referred to as Upregulated in Infectious Sporozoites gene 4 (UIS4)
UIS4 Upregulated in Infectious Sporozoites gene 4
ETRAMP10.3 is not a functional ortholog of UIS4 and may play a different biological role. Although the biological function of ETRAMP10.3 has not yet been completely resolved, localization to vesicular structures in the host erythrocyte suggests a role in host-parasite interaction or in remodeling of infected erythrocyte. ETRAMP10.3 appears to play a key role in the Plasmodium life cycle. When ETRAMP10.3 is deleted, the deletion can lead to the disruption of liver-stage development in mice and asexual blood stage progression.
UIS4 and ETRAMP10.3 are in the literature sometimes used to refer to different proteins, in context of the present disclosure, the terms “UIS4* and "ETRAMP10.3” interchangeably to refer to ETRAMP10.3.
Plasmodium UIS4 sequences are known (see, e.g., UniProt accession number Q8DM9, which is incorporated herein by reference in its entirety).
An exemplary UIS4 amino add sequence is provided in Table 2.
Liver Specific Protein 1 fUSP-1 Liver Specific Protein 1 fUSP-1
LISP-1 Liver specific protein 1
PVM parasitophorous vacuolar membrane
Intracellular Plasmodium deficient In LISP-1 develop into hepatic merozoites and display normal infectivity to erythrocytes (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep; 11(9): 1329-1339, which is incorporated herein by reference in its entirety).
USPl-defident liver-stage Plasmodium do not rupture PVM and remain trapped inside hepatocytes (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep; 11(9): 1329-1339, which is incorporated hereto by reference in its entirety).
Plasmodium LISP-1 sequences are known (see, e.g., UniProt accession number A0A210C2X6, Q8ILR5, each of which is incorporated herein by reference in its entirety).
An exemplary LISP-1 amino add sequence is provided In Table 2.
Liver spedfic protein 2 contains a modified 6-cys domain and is expressed during Plasmodium development in hepatocytes (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(l):66-79, which is incorporated herein by reference In Its entirety). LISP-2 was shown to be expressed by liver stages Plasmodium, exported to hepatocytes, and be distributed throughout the host cell, including the nucleus (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(l): 66-79, which is incorporated herein by reference In Its entirety).
Plasmodium LISP-2 sequences are known (see, e.g., UniProt accession number A0A2I0BZR4, Q8I1X6, Q9U0D4, each of which is Incorporated hereto by reference in its entirety).
An exemplary LISP-2 amino add sequence is provided in Table 2.
Thrombospondin-related adhesion protein contains an N-terminal domain that is commonly referred to as von Willebrand factor A domain, although it is most similar to an integrin I domain because it contains a metal Ion-dependent adhesion site (MIDAS) with a bound Mg 2t Ion that Is required for sporozoite motility In vitro and infection in vivo (see, e.g., Lu et al., PLoS One. 2020; 15(1): e0216260, which is incorporated herein by reference in its entirety).
MIDAS metal Ion-dependent adhesion site
the I domain is inserted in an extensible p-ribbon and followed by a thrombospondin repeat (TSR) domain, a proline-rich segment at the C-terminus, a single-pass transmembrane domain, and a cytoplasmic domain (see, e.g., Lu et al., PLoS One. 2020; 15(1): e0216260, which is incorporated herein by reference in its entirety).
TSR thrombospondin repeat
TRAP is stored in the mkronemes and becomes surface exposed at the sporozoite anterior tip when parasite comes in contact with host cells (Akhouri et al., Malar J. 2008 Apr 22;7:63. doi: 10.1186/1475-2875-7-63, which is incorporated herein by reference in its entirety). TRAP also plays an important role in liver cell invasion of sporozoites by helping sporozoites in gliding motility and in recognition of host receptors on the mosquito salivary gland and hepatocytes (Akhouri et al., Malar J. 2008 Apr 22;7:63. doi: 10.1186/1475-2875-7-63, which is incorporated hereto by reference in its entirety).
PlasmodiumTRAP sequences are known (see, e.g., UniProt accession numbers A0A5Q2EXK8, A0A5Q2EZD7, A0A5Q2F1F6, A0A5Q2F2B8, A0A5Q2F2H6, A0A5Q2F4G9, 076110, P16893, Q01507, Q26020, Q76NM2, W8VNB6, each of which is Incorporated herein by reference in its entirety), and an exemplary TRAP amino add sequence Is provided In Table 2.
LSAP-1 Liver-stage-associated protein
salivary gland sporozoites see, e.g., Siau et al., PLoS Pathog. 2008 Aug 8;4(8):el000121, which Is Incorporated herein by reference In Its entirety.
LSAP-1 is among the most abundant transcripts In the salivary gland transcriptome but has not been detected in proteomlc surveys of sporozoites. Rather, expression has only been detected only In liver stages (see, e.g., Slau et al., PLoS Pathog. 2008 Aug 8;4(8):el000121, which is Incorporated herein by reference In its entirety).
Plasmodium LSAP-1 sequences are known (see, e.g., UniProt accession number Q8I632, W7JR53, each of which Is incorporated herein by reference in Its entirety).
An exemplary LSAP-1 amino add sequence is provided In Table 2.
LSAP-2 Is also among the most abundant transcripts In the salivary gland transcriptome but has not been detected in proteomlc surveys of sporozoites. LSAP-2 has shown some efficacy as a vaccine when combined with other antigens. See, e.g., Halbroth et al., Infect Immun. 2020 Jan 22;88(2):e00573-19. doi: 10.1128/IAI.00573-19. Print 2020 Jan 22, which Is incorporated herein by reference In its entirety.
Plasmodium LSAP-2 sequences are known (see, e.g., UniProt accession number Q8I632, W7JR53, each of which is incorporated herein by reference in its entirety).
An exemplary LSAP-2 amino add sequence is provided in Table 2.
LSA-1 Liver-Staae Antigen 1
LSA-1 Liver-Stage Antigen 1
the function of LSA-1 remains currently not known (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vacdne Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), Cun-ent Topics in Malaria, IntechOpen, London. 10.5772/65592, which is incorporated herein by reference in its entirety).
LSA-1 is a 230 kDa preerythrocytic stage protein containing a large central region consisting of over eighty 17 amino add residue repeat units flanked by highly conserved C- and N-termlnal regions (Richie, T.L. and Parekh, F.K. (2009) Malaria, which is incorporated herein by reference in its entirety). In Vaccines for Biodefense and Emerging and Neglected Diseases (Barrett, A.D.T. and Stanberry L.R., eds), pp. 1309-1364, Elsevier, which is incorporated herein by reference in its entirety). LSA1 is expressed only by liver stage Plasmodium and not by sporozoites (Richie, T.L.
Plasmodium LSA-1 sequences are known (see, e.g., UniProt accession number Q25886, Q25887, Q25893, Q26028, Q9GTX5, 096125, each of which Is Incorporated herein by reference In Its entirety).
An exemplary LSA-1 amino acid sequence is provided in Table 2.
LSA-3 Liver stage antigen 3
LSA-3 Liver stage antigen 3 Is a 200-kDa protein that Is composed of three nonrepeating regions (NR- A, NR-B, and NR-C) flanking two short repeat regions and one long repeat region (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), which Is incorporated herein by reference in its entirety), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, which is incorporated herein by reference in its entirety).
the nonrepeat regions are well conserved across geographically diverse strains of Plasmodium faidparum (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), Current Topics In Malaria, IntechOpen, London. 10.5772/65592, which is Incorporated herein by reference in Its entirety).
the most significant variation Is In the repeating regions due to organization and number of repeating subunits rather than composition of the repeating regions (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates In Malaria', in A. J. Rodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, which is Incorporated herein by reference In its entirety).
Plasmodium LSA-3 sequences are known (see, e.g., UniProt accession number C7DU21, C7DU22, C7DU23, C7DU24, C7DU25, C7DU26, C7DU27, C7DU28, C7DU29, C7DU32, C7DU33, C7DU34, C7DU36, C7DU37, C7DU38, C7DU39, C7DU40, Q8I042, Q8I0A5, Q8I0D0, Q8IFR1, Q8IFR2, Q8IFR3, Q8IFR4, Q8IFR5, Q8IFR6, Q8IFR7, Q8IFR8, Q8IFR9, Q8IFS0, Q8IFS1, Q8IFS2, Q8IFS3, Q8IFS4, Q8IFS5, Q8IFS6, Q8IFS7, Q8IFS8, Q8IFS9, Q8IFT0, Q8IFT1, Q8IFT2, Q8IFT3, Q8IFS
Glutamic add-rich protein is a 80kDA protein which derives its name from its glutamic rich amino add sequence which comprises 24% of all its residues. GARP Is predominantly expressed in ring stages and trophozoites and has been shown to be a non-essential gene in cell culture but highly immunogenic in animal models. Although GARP is non-essential in cell culture, its localization to the periphery of infected erythrocytes may indicate a role in the sequestration of infected erythrocytes. CARP'S involvement in sequestration has been proposed to occur by way of binding with an chloride/bicarbonate anion exchanger. Antibodies against GARP have been proposed to serve as signatures of protection against severe malaria and have shown efficacy in experimental trials
EXP1 (3D7) MKJLSVFFLALFHIFNKESLAEKTNKGTCSGVSSKKKNKKGSGEPUDVHDUSDMIKKEE ELVEVNKRKSKYKLATSVLAGLLGWSTVLLGGVGLVLYNTEKGRHPFKIGSSDPADNANP DADSESNGEPNAGPQVTAQDVTPEQPQGDDNNLVSGTEH _
EXP2 (3D7) MKVSYIFSFFLLFFVYKNTNTWCDNGYGDLAATSALTTVIKDPISLTIKDIYEHGVKNPFT KIIHKLKKHRYRKVLRWSRMWWVLLVREIVGDNnEKKTEKALREIWDQCnAVYNNTL NAVESKPLLFLHGILNECRNNFATKLRQDPSUVAKIDQIIKSQIYRFWVSEPYLKIGRSHT LYTHrrPDAVPQLPKECTlKHLSSYMEEKLKSMESKKNIESGKYEFDVDSSETDSTKDDG KPDDDDDDDDNFDDDDNFDDDTVEEEDASGDLFKNEKKDENKE _
RNA technologies as a modality to express one or more Plasmodium polypeptide construct that includes one or more malarial proteins, or one or more portions thereof, described herein.
a Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions of one or more Plasmodium Rh5 Invasion complex polypeptides).
one or more polypeptides or portions thereof of a Plasmodium Rh5 invasion complex can include one or more polypeptides or portions thereof of a Plasmodium reticulocyte-binding protein homolog 5 (Rh5), a Plasmodium Cysteine-Rjch Protective Antigen (CyRPA), a Plasmodium Rh5-lnteracting Protein (Rlpr), a Plasmodium P 113, a Plasmodium thrombospondin-related apical merozoite protein (TRAMP), and/or a Plasmodium cysteine-rich small secreted protein (CSS).
Rh5 Plasmodium reticulocyte-binding protein homolog 5
CyRPA Plasmodium Cysteine-Rjch Protective Antigen
Rhpr Plasmodium Rh5-lnteracting Protein
TRAMP Plasmodium thrombospondin-related apical merozoite protein
CCSS Plasmodium
a portion of a Rh5 invasion complex polypeptide can be a characteristic portion of a Rh5 invasion complex polypeptide.
a Plasmodium polypeptide construct additionally Includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multimerization region, and/or a linker, as described herein.
a Plasmodium polypeptide construct described herein includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprise one or more Plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof.
a Plasmodium polypeptide construct described herein includes one or more polypeptides or antigenic portions of an Rh5 polypeptide, e.g., Plasmodium Rh5 polypeptide, e.g., P. fafciparum Rh5 (SEQ ID NO: 1), or a variant thereof (e.g., a glycosylation variant).
an Rh5 polypeptide may include a Plasmodium Rh5 N-terminal disordered domain, a first Plasmodium Rh5 ordered domain, a Plasmodium Rh5 linking disordered domain, a second Plasmodium Rh5 ordered domain (e.g., a C-terminal Plasmodium Rh5 ordered domain), or a combination thereof.
a portion of Rh5 (or Rh5 polypeptide portion) (e.g., an antigenic portion of Rh5 or Rh5 antigenic portion) may refer to parts of an Rh5 polypeptide domain or parts spanning two or more Rh5 polypeptide domains.
a Plasmodium polypeptide construct comprises one or more antigenic portions of Rh5.
one or more antigenic portions of Rh5 comprise one or more Rh5 ordered domains.
a Plasmodium Rh5 polypeptide or antigenic portion thereof comprises 25, 30, 35, 40, or 45 contiguous amino adds of an Rh5 ordered domain, or a variant thereof (e.g., a glycosylation variant).
a Plasmodium Rh5 antigenic portion comprises an Rh5 ordered domain that corresponds to amino adds 140-247 or amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof (e.g., glycosylation variants).
a Plasmodium Rh5 antigenic portion comprises an Rh5 ordered domain that corresponds to amino acids 297-526 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof (e.g., a glycosylation variant).
one or more antigenic portions of Rh5 comprise (i) an Rh5 ordered domain that corresponds to amino adds 140-247 or amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and/or (ii) an Rh5 ordered domain that corresponds to amino adds 297-526 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof.
one or more Plasmodium Rh5 antigenic portions comprise one or more unpaired cysteines. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two unpaired cysteines, in some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a cysteine at position 203, 329, or both, as numbered according to SEQ JD NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, 329, or both, as numbered according to SEQ ID NO: 1.
one or more Plasmodium Rh5 antigenic portions comprise one, two, three, or four N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution at one or more N-linked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution.
one or more Piasmocfium Rh5 antigenic portions comprise an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Piasmodkim Rh5 antigenic portion comprises a Rh5 ordered domain that corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) (or amino adds 140-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino add substitutions).
an Rh5 ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203.
an Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N214. In some embodiments, an Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino add substitution that prevents glycosylation at position 214. In some embodiments, an amino add substitution that prevents glycosylation at position 214 comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino add substitution prevents glycosylation at position 214 comprises a NX[T/S] to NXA substitution.
a Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214.
an Rh5 antigenic portion (e.g., a Rh5 ordered domain) comprises a Plasmepsin X (PMX) deavage site.
PMX is a conserved aspartic protease that is responsible for proteolytic processing and activation of proteins and other proteases involved in life cyde progression and/or development of Plasmodium.
a PMX cleavage site of an Rh5 ordered domain comprises a sequence of FLQY (SEQ ID NO: 36), where cleavage occurs between L and Q.
a cleaved Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or a variant thereof.
an Rh5 antigenic portion e.g., an Rh5 ordered domain
a Plasmepsin X (PMX) deavage site does not comprise a Plasmepsin X (PMX) deavage site.
a Plasmodium Rh5 antigenic portion comprises a Rh5 ordered domain that corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) (or amino adds 146-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino add substitutions).
an Rh5 ordered domain corresponds to amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203.
an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203.
an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N214.
an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino add substitution that prevents glycosylation at position 214.
an amino add substitution that prevents glycosylation at position 214 comprises a NX[T/S] to QX[T/S] substitution.
amino add substitution prevents glycosylation at position 214 comprises a NX[T/S] to NXA substitution.
an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214.
an Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 329 and/or glutamine at position 297.
an Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and conyrises a cysteine at position 329 and asparagine at position 297.
a Plasmodium Rh5 polypeptide or antigenic portion thereof comprises an Rh5 disordered domain or antigenic portion thereof.
an Rh5 disordered domain is an Rh5 N- terminal disordered domain that corresponds to amino adds 26-139 of a wild-type Rh5 (SEQ ID NO: 1).
an Rh5 disordered domain is an Rh5 linking disordered domain that corresponds to amino adds 248- 296 of a wild-type Rh5 (SEQ ID NO: 1).
one or more Plasmodium Rh5 antigenic portions comprise two Rh5 ordered domains and one Rh5 disordered domain. In some embodiments, one or more Plasmodium Rh5 antigenic portions correspond to amino adds 140-526 of a wild-type Rh5 (SEQ ID NO: 1). In some embodiments, one or more Piasmocfium Rh5 antigenic portions correspond to amino adds 26-247 and 297-526 of a wild-type Rh5 (SEQ ID NO: 1).
a Plasmodium polypeptide construct does not include an Rh5 disordered domain.
a Plasmodium Rh5 polypeptide or antigenic portion thereof does not comprise an Rh5 disordered domain that corresponds to amino adds 26-139 or amino acids 248-296 of a wiki-type Rh5 (SEQ ID NO: 1).
a Plasmodium polypeptide construct does not Indude an Rh5 secretory signal, e.g., does not indude a sequence corresponding to amino adds 1-25 of a wild-type Rh5 (SEQ ID NO: 1).
RhS secretory signal SEQ ID NO: 132
a Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof comprising two Rh5 ordered domains. In some embodiments, a Plasmodium polypeptide construct comprises two Plasmodium Rh5 antigenic portions, where each antigenic portion comprises an Rh5 ordered domain. In some embodiments, a Plasmodium polypeptide construct comprises two Rh5 ordered domains that correspond to (I) amino adds 140-247 or 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and (II) amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof. In some embodiments, a Plasmodium polypeptide construct comprises two Rh5 ordered domains that are directly adjacent to one another.
a Plasmodium polypeptide construct comprises an Rh5 polypeptide comprising exactly two antigenic portions of Rh5, where the two antigenic portions comprise or consist of two Rh5 ordered domains.
two Rh5 ordered domains correspond to (i) amino acids 140-247 or amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and (II) amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof.
a Plasmodium polypeptide construct comprising two Rh5 ordered domains, where a first Rh5 ordered domain corresponds to amino adds 140-247 or amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214, and where a second Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 329 and/or glutamine at position 297.
a Plasmodium polypeptide construct comprising two Rh5 ordered domains, where a first Rh5 ordered domain corresponds to amino acids 140-247 or amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214, and where a second Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 329 and asparagine at position 297.
a Plasmodium polypeptide construct does not include a Rh5 disordered domain (e.g., does not indude amino adds 26-139 or amino adds 248-296 of a wild-type Rh5 (SEQ ID NO: 1). In some embodiments, a Plasmodium polypeptide construct does not include a Rh5 secretory signal, e.g., does not indude a sequence corresponding to amino adds 1-25 of a wild-type Rh5 (SEQ ID NO: 1).
a Plasmodium polypeptide construct comprises one or more antigenic portions of Plasmodium Rh5 comprising a PMX deavage site. In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Plasmodium Rh5 comprising a PMX cleavage site that comprises or consists of an amino add sequence of NFLQ (SEQ ID NO: 189).
a Plasmodium polypeptide construct described herein Includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprise one or more Plasmodium Cysteine-Rich Protective Antigen (CyRPA) polypeptides or antigenic portions thereof.
a Plasmodium polypeptide construct described herein includes one or more regions or portions (e.g., antigenic portions) of a CyRPA, e.g., Plasmodium CyRPA, e.g., P.
falciparum CyRPA (SEQ ID NO: 3), or a variant thereof (e.g., one or more antigenic portions of a CyRPA, e.g., Plasmodium CyRPA, e.g., P. falciparum CyRPA).
a Plasmodium polypeptide construct comprises an antigenic portion of CyRPA.
a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises 25, 30, 35, 40, or 45 contiguous amino adds of CyRPA, or a variant thereof (e.g., a glycosylation variant).
a Plasmodium CyRPA antigenic portion does not comprise an endogenous signal sequence.
a Plasmodium CyRPA antigenic portion is operably linked to a heterologous signal sequence, as further described herein.
a Plasmodium CyRPA antigenic portion comprises or consists of amino adds 30-362 of a wild-type CyRPA (SEQ ID NO: 3) or a variant thereof (e.g., a glycosylation variant).
a Plasmodium CyRPA antigenic portion comprises or consists of a sequence according to SEQ ID NO: 133 or a variant thereof (e.g., a glycosylation variant).
a Plasmodium CyRPA antigenic portion comprises or consists of a sequence with at least 85% sequence identity to SEQ ID NO: 133 or a variant thereof (e.g., a glycosylation variant).
a Plasmodium CyRPA antigenic portion comprises or consists of amino acids 29- 362 of a wild-type CyRPA (SEQ ID NO: 3) or a variant thereof (e.g., a glycosylation variant).
a Plasmodium CyRPA antigenic portion comprises or consists of a sequence according to SEQ ID NO: 169 or a variant thereof (e.g., a glycosylation variant).
a Plasmodium CyRPA antigenic portion comprises or consists of a sequence with at least 85% sequence identity to SEQ ID NO: 169 or a variant thereof (e.g., a glycosylation variant).
one or more Plasmodium CyRPA antigenic portions comprise one or more N- llnked glycosylation sites.
a Plasmodium CyRPA antigenic portion comprises one, two, or three N-linked glycosylation sites.
a Plasmodium CyRPA antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, where the amino acid substitution prevents glycosylation.
an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution.
amino acid substitution prevents glycosylation comprises a NX[T/S) to NXA substitution.
a Plasmodium CyRPA antigenic portion comprises two amino acid substitutions at N-linked glycosylation sites that prevent glycosylation. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium CyRPA antigenic portion comprises an asparagine at position 145, position 322, position 338, or a combination thereof, as numbered according to SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, position
a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, and position 338, as numbered according to SEQ ID NO: 3.
a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 134.
a Plasmodium CyRPA antigenic portion comprises an asparagine at position 146, position 323, position 339, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 146, position 323, position
a Plasmodium CyRPA antigenic portion comprises a glutamine at position 146, position 323, and position 339, as numbered according to SEQ ID NO: 3.
a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 170.
a Plasmodium polypeptide construct described herein includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprise one or more Plasmodium P113 polypeptides or antigenic portions thereof.
a Plasmodium polypeptide construct described herein includes one or more regions or portions (e.g., antigenic portions) of a P113, e.g., Plasmodium P113, e.g., P. falciparum P113 (SEQ ID NO: 6), or a variant thereof (e.g., one or more antigenic portions of a P113, e.g., Plasmodium P113, e.g., P. falciparum P113).
a Plasmodium polypeptide construct comprises one or more antigenic portion of P113.
a Hasmodum P113 polypeptide or antigenic portion thereof comprises 25, 30, 35, 40, or 45 contiguous amino acids of a Pl 13 polypeptide, or a variant thereof (e.g., a glycosylation variant).
a Plasmodium polypeptide construct comprises an antigenic portion of P113.
a Plasmodium P113 antigenic portion does not comprise an endogenous signal sequence. In some embodiments, a Plasmodium P113 antigenic portion is operably linked to a heterologous signal sequence, as described further herein. In some embodiments, a Plasmodium Pl 13 antigenic portion does not comprise a GPI anchor site. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of amino acids 23-939 of a wild-type P113 (SEQ ID NO: 6) or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of a sequence according to SEQ ID NO: 135. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 135.
a Plasmodium P113 antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium P113 antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a PlasnxxSum P113 antigenic portion comprises an amino acid substitution at one or more N-llnked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an amino add substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino add substitution prevents glycosylation comprises a NX(T/S] to NXA substitution.
a Plasmodium P113 antigenic portion comprises two, three, four, five, six, or seven amino acid substitutions at N-linked glycosylation sites that prevent glycosylation. In some embodiments, a Plasmodium P113 antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
a Plasmodium P113 antigenic portion comprises an asparagine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.
a Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.
a Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, and position 854, as numbered according to SEQ ID NO: 6.
a Plasmodium P113 antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 136.
Plasmodium polypeptide construct described herein includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprises one or more Plasmodium Rh5-interacting Protein (Ripr) polypeptides or antigenic portions thereof.
a Plasmodium polypeptide construct described herein includes one or more regions or portions (e.g., antigenic portions) of a Ripr, e.g., Plasmodium Ripr, e.g., P. falciparum Ripr (SEQ ID NO: 2), or a variant thereof (e.g., one or more antigenic portions of a Ripr, e.g., Plasmodium Ripr, e.g., P. falciparum Ripr).
an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 178.
an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 178. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 177. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 177. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 176.
an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 176. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 175. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 175. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 174.
an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 174. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 173. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 173. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 172.
an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 172. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 171. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 171.
an Plasmodium Ripr antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium Ripr antigenic portion comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve N-linked glycosylation sites. In some embodiments, a Plasmodium Ripr antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution.
amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution.
a Plasmodium Ripr antigenic portion comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
an antigenic portion of a Plasmodium Ripr polypeptide comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.
an antigenic portion of a Plasmodium Ripr polypeptide comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.
an antigenic portion of a Plasmodium Ripr polypeptide comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2.
the antigenic portion of a Plasmodium Ripr comprises or consists of an amino add sequence according to SEQ ID NO: 173.
an antigenic portion of a Plasmodium Ripr polypeptide comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered accordng to SEQ ID NO: 2.
an antigenic portion of a Pfasmodium Ripr polypeptide comprises a ⁇ utamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.
an antigenic portion of a Plasmodium Ripr polypeptide comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2.
an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of SEQ ID NO: 172.
a Plasmodium Ripr antigenic portion comprises a PMX cleavage site.
a PMX cleavage site of a Ripr antigenic portion comprises a sequence of GNISMLEIQNEE (SEQ ID NO: 37).
a Plasmodium Ripr antigenic portion does not comprise a PMX cleavage site.
a PMX cleavage site of a Ripr antigenic portion comprises or consists of a sequence of SMLE (SEQ ID NO: 191).
an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 179.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises 25, 30, 35, 40, or 45 contiguous amino acids of a TRAMP polypeptide, or a variant thereof (e.g., a glycosylation variant).
a Plasmodium polypeptide construct comprises an antigenic portion of TRAMP.
a Plasmodium TRAMP antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium TRAMP antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a PlasmodiumTRMAP antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, an amino add substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitutlon. In some embodiments, amino add substitution prevents glycosylation comprises a NX[T/S] to NXA substitution.
a Plasmodium TRAMP antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 4. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 181.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 4.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 180.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a PMX cleavage site.
a PMX cleavage site of TRAMP antigenic portion comprises or consists of a sequence of HFLQ (SEQ ID NO: 192).
an antigenic portion of a PlasmodkimlRMAP polypeptide comprises a SUB2 cleavage site.
a SUB2 cleavage site of TRAMP antigenic portion comprises or consists of ILMIILPIVLVISimLYHIFY (SEQ ID NO: 193).
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a transmembrane region (also referred to as a "transmembrane domain”).
an antigenic portion of a PiasmodiumTWM? polypeptide comprises a transmembrane region at the C-terminus.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region.
an antigenic portion of a Plasmodium TRAMP polypeptide comprises a transmembrane region that comprises or consists of an HSV gD transmembrane region, e.g., comprising or consisting of an amino add sequence of SEQ ID NO:75.
a Plasmodium TRAMP polypeptide does not comprise a transmembrane region.
a Plasmodium polypeptide construct described herein Includes one or more Plasmodium Rh5 Invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprises one or more Plasmodium cysteine-rich small secreted protein (CSS) polypeptides or antigenic portions thereof.
a Plasmodium polypeptide construct described herein includes one or more regions or portions of a CSS, e.g., Plasmodium CSS, e.g., P.
falciparum CSS SEQ ID NO: 5 or 214
a variant thereof e.g., one or more antigenic portions of a CSS, e.g., Plasmodium CSS, e.g., P. falciparum CSS").
an antigenic portion of a Plasmodium CSS polypeptide comprises a C30S mutation, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a serine at position 30, as numbered according to SEQ ID NO: 5 or 214. [0483] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 194.
an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 182. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 182.
an antigenic portion of a Plasmodium CSS polypeptide comprises 25, 30, 35, 40, or 45 contiguous amino acids of a CSS polypeptide, or a variant thereof (e.g., a glycosylation variant).
a Plasmodium polypeptide construct comprises one or more antigenic portions of CSS.
a Plasmodium CSS antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium CSS antigenic portion comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, a Plasmodium CSS antigenic portion comprises an amino add substitution at one or more N-llnked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, a Plasmodium CSS antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.
an antigenic portion of Plasmodium CSS polypeptide comprises a serine at position 80, as numbered according to SEQ ID NO: 5 or 214.
an antigenic portion of a Plasmodium CSS polypeptide comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, and position 88, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 184.
an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 185.
an antigenic portion of a Plasmodium CSS polypeptide comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214.
an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214.
an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 183.
a Plasmodium polypeptide construct described herein Includes a secretory signal, e.g., that Is functional In mammalian cells.
a utilized secretory signal is a heterologous secretory signal.
a heterologous secretory signal comprises or consists of a non-human secretory signal.
a heterologous secretory signal comprises or consists of a viral secretory slgnal.
a viral secretory signal comprises or consists of an hSV secretory signal (e.g., an HSV- 1 or HSV-2 secretory signal).
an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal.
a secretory signal Is characterized by a length of about 15 to 30 amino adds.
a secretory signal is positioned at the N-terminus of a Plasmodium polypeptide construct described herein.
a secretory signal preferably allows transport of a Plasmodium polypeptide construct with which it is assodated into a defined cellular compartment, preferably a cell surface, endoplasmic reticulum (ER) or endosomal-lysosomal compartment [0500]
a secretory signal comprises or consists of a Plasmodium secretory signal.
a Plasmodium seaetory signal comprises or consists of a Plasmodium CSP secretory signal (e.g., from Plasmodium fafdpanim, e.g., from Plasmodium falciparumVsfitete 3D7 (SEQ ID NO: 7)).
a Plasmodium seaetory signal comprises a Plasmodium Rh5 seaetory signal.
a Plasmodium secretory signal comprises a Plasmodium CyRPA secretory signal.
a Plasmodium sec.xe.torf signal comprises or consists of an amino acid sequence according to SEQ ID NO: 187.
a seaetory signal is selected from an S1S2 secretory signal (aa 1-19), an Immunoglobulin secretory signal (aa 1-22), a human SPARC secretory signal, a human Insulin Isoform 1 seaetory signal, a human albumin secretory signal, etc.
S1S2 secretory signal aa 1-19
an Immunoglobulin secretory signal aa 1-22
a human SPARC secretory signal e.g., a human Insulin Isoform 1 seaetory signal
a human albumin secretory signal etc.
a Plasmodium polypeptide construct described herein does not comprise a secretory signal.
a seaetory signal is one listed in Table 3, or a secretory signal having 1, 2, 3, 4, or 5 amino acid differences relative thereto.
a signal sequence is selected from those Included In the Table 3 below and/or those encoded by the sequences In Table 4 below.
a Plasmodium polypeptide construct described herein Includes a transmembrane region (also referred to as a "transmembrane domain”).
a transmembrane region is located at the N-termlnus of a Plasmodium polypeptide construct.
a transmembrane region Is located at the C-terminus of a Plasmodium polypeptide construct.
a transmembrane region is not located at the N-terminus or C-terminus of a Plasmodium polypeptide construct.
a heterologous transmembrane region does not comprise a hemagglutin transmembrane region.
a heterologous transmembrane region comprises or consists of a non-human transmembrane region.
a heterologous transmembrane region comprises or consists of a viral transmembrane region.
a heterologous transmembrane region comprises or consists of an HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region.
an HSV transmembrane region comprises or consists of an HSV gD transmembrane region, e.g., comprising or consisting of an amino add sequence of GLIAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR (SEQ ID NO: 75).
a heterologous transmembrane region comprises or consists of a human transmembrane region.
a human transmembrane region comprises or consists of a human decay accelerating factor glycosylphosphatidylinositol (hDAF-GPI) anchor region.
an hDAF- GPI anchor region comprises or consists of an amino add sequence of PNKGSGTTSGTTRLi-SGHTCFTLTGLLGTLVTMGLLT (SEQ ID NO:76).
a transmembrane region comprises or consists of a Plasmodium transmembrane region.
a utilized transmembrane region Is one that is normally associated with CSP In nature.
a Plasmodium transmembrane region comprises or consists of a Plasmodium QSP glycosylphosphatidylinositol (GPI) anchor region.
a Plasmodium CSP GPI anchor region is from Plasmodium falciparum.
a Plasmodium CSP GPI anchor region is from Plasmodium falciparum isolate 3D7 (SEQ ID N0:7), e.g., amino adds 374-397 of SEQ ID N0:7.
a transmembrane region comprises PlasmodiumTRAP transmembrane region. In some embodiments, a transmembrane region comprises Plasmodium P113 transmembrane region.
a utilized transmembrane region is a heterologous transmembrane region.
a Plasmodium polypeptide construct described herein does not comprise a transmembrane region.
a Plasmodium polypeptide construct described herein includes one or more multimerization regions (e.g., a heterologous multimerization region).
a heterologous multimerization region comprises a dimerization, trimerizatlon or tetramerization region.
a multimerization region is one described in W02017/081082, which is Incorporated herein by reference In Its entirety (e.g., SEQ ID NOs: 1116-1167, or fragments or variants thereof).
Exemplary trimerizatlon and tetramerization regions include, but are not limited to, engineered leucine zippers, fibritki fbldon domain from enterobacteria phage T4, GCN4pll, GCN4-pll, and p53.
a provided Plasmodium polypeptide construct described herein Is able to form a trimeric complex.
a provided Plasmodium polypeptide construct may comprise a multimerization region allowing formation of a multimeric complex, such as for example a trimeric complex of a Plasmodium polypeptide construct described herein.
a multimerization region allowing formation of a multimeric complex comprises a trimerization region, for example, a trimerizatlon region described herein.
a Plasmodium polypeptide construct includes a T4-fibritin-derived '"fbldon"' trimerization region, for example, to increase its immunogenicity.
a Plasmodium polypeptide construct includes a multimerization region comprising or consisting of the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 78).
a Plasmodium polypeptide construct described herein includes one or more linkers.
a linker is or comprises 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids.
a linker is or comprises no more than about 30, 25, 20, 15, 10 or fewer amino adds.
a linker can indude any amino acid sequence and is not limited to any particular amino adds.
a linker comprises one or more gtydne (G) amino acids.
a linker comprises one or more serine (S) amino acids.
a linker comprises a glycine-serlne linker.
a “glydne-serlne inker” as used herein refers to a linker that comprises predominantly (e.g., 80% or more) glyche end serine amino acids.
a linker indudes amino adds selected based on a deavage predictor to generate highly-deavable linkers.
a linker is or comprises S-G4-S-G4-S.
a linker Is or comprises GSPGSGSGS (SEQ ID NO: 79).
a linker is or comprises GGSGGGGSGG (SEQ ID NO: 80).
a linker Is one presented in Table 6.
a linker is or comprises a sequence as set forth in W02017/081082, which Is incorporated herein by reference In Its entirety (see SEQ ID NOs: 1509-1565, or a fragment or variant thereof).
a Plasmodium polypeptide construct described herein comprises a linker between a C-terminal region or portion thereof and a transmembrane region. In some embodiments, a Plasmodium polypeptide construct described herein comprises a linker after a minor repeat sequence.
a Plasmodium polypeptide construct described herein includes one or more selfassembling regions (e.g., a self-assembling nanopartide region, e.g., a heterologous self-assembling nanoparticle region).
a self-assembling nanoparticle region is a ferritin region.
a ferritin region is from H. pylori.
a ferritin region comprises or consists of a sequence of: DIIKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEHAKKUIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYE HEQHISESINNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIEUGNENHGLYLADQYVKGIAKSRKS (SEQ ID NO: 88).
one or more tags may be used when designing and testing constructs (e.g., Plasmodium polypeptide constructs described herein) in certain contexts (e.g., in vitro, ex vivo, etc.).
one or more tags may be directly connected to a Plasmodium polypeptide construct through a peptide bond (e.g., at the S'-end or 3'-end of a construct).
one or more tags may be connected to a Plasmodium polypeptide construct through one or more linkers (e.g., one or more linkers described herein).
one or more tags may be internally embedded within a Plasmodium polypeptide construct, wherein the one or more tags are directly connected to the construct through peptide bonds (e.g., at the S'-end and 3'-end of the one or more tags).
one or more tags may be Internally embedded within a Plasmodium polypeptide construct, wherein the one or more tags are connected to the construct through one or more linkers (e.g., one or more linkers described herein).
a Plasmodium polypeptide construct described herein Includes one or more tags.
a Plasmodium polypeptide construct described herein Includes one or more detection tags (e.g., HiBit tag, HA tag, etc.).
a Plasmodium polypeptide construct described herein indudes a tag that comprises or consists of a HiBit tag.
a HiBit tag has an amino add sequence according to SEQ ID NO: 139.
Plasmodium polypeptide construct described herein includes one or more
a Plasmodium polypeptide construct described herein Includes one or more regions or portions of a Rh5 invasion complex polypeptide from Plasmodium fatoparum, preferably from Plasmodium fatoparum isolate 3D7.
a Rh5 Invasion complex polypeptide is selected from: Plasmodium Rh5, Plasmodium CyRPA, Plasmodium Ripr, Plasmodium P113, Plasmodium TRAMP, and Plasmodium CSS.
the Plasmodium polypeptide construct described herein Includes an ordered domain of Rh5 from Plasmodium fatoparum, preferably from Plasmodium faidparum isolate 3D7. In some embodiments, the Plasmodium polypeptide construct described herein includes an ordered domain of Rh5 from Plasmodium faidparum, preferably from Plasmodium fatoparum isolate 3D7 (referred to herein as a "Rh5 construct” or "Rh5 ordered domain”).
an ordered domain of Rh5 comprises the amino add sequence of positions 140- 247 of SEQ ID NO: 1, or an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 140-247 of SEQ ID NO: 1.
an ordered domain of Rh5 comprises the amino add sequence of positions 140-247 of SEQ ID NO: 1, or the amino add sequence of positions 140-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
an ordered domain of Rh5 comprises the amino acid sequence of positions 146- 247 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 146-247 of SEQ ID NO: 1.
an ordered domain of Rh5 comprises the amino add sequence of positions 146-247 of SEQ ID NO: 1, or the amino acid sequence of positions 146-247 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.
an ordered domain of Rh5 comprises the amino acid sequence of positions 297- 526 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 297-526 of SEQ ID NO: 1.
an ordered domain of Rh5 comprises the amino add sequence of positions 297-526 of SEQ ID NO: 1, or the amino add sequence of positions 297-526 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.
an ordered domain of Rh5 can have the following structure: sequence corresponding to amino adds 140-247 of SEQ ID NO: 1 (140-247 Ordered Domain); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 (140- 247 Ordered Domain-203Y); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (140-247 Ordered Domain-A214gly); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (140-247 Ordered Domain-203Y-A214gly); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a glutamine at position 214 (140-247 Ordered Domaln-N214Q); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a ty
a Plasmodium polypeptide construct described herein includes two ordered domains of Rh5.
two ordered domains of Rh5 are from Plasmodium fakjparum, preferably from Plasmodium fakjparum isolate 3D7.
a Plasmodium polypeptide construct comprises a Rh5 polypeptide or antigenic portion thereof that includes 2 ordered domains of Rh5.
an Rh5 portion of a malarial polypeptide construct includes two ordered domains of Rh5 that have the following structure:
Plasmodium polypeptide construct indudes an Rh5 polypeptide or antigenic portion thereof that Indudes 2 ordered domains of Rh5 and a linker.
an Rh5 portion of a maliarlal polypeptide has the following structure:
Plasmodium polypeptide construct includes an Rh5 polypeptide or antigenic portions thereof that includes 2 ordered domains of Rh5 and one or more disordered domains.
a disordered domain is an L-disordered domain of Rh5.
an L-disordered domain comprises a sequence corresponding to amino acids 248-296 of SEQ ID NO: 1.
a disordered domain is an L-disordered domain of Rh5.
an N-disordered domain comprises a sequence corresponding to amino adds 26-139 of SEQ ID NO: 1.
an Rh5 portion comprises both an L-disordered domain and an N-disordered domain of Rh5.
an Rh5 portion of a malarial polypeptide has two ordered domains and one or two disordered domains of Rh5.
an Rh5 portion of a malarial polypeptide has the following structure:
N-Disordered Domain 140-247 Ordered Domain-203Y - 297-526 Ordered Domain;
N-Disordered Domain 140-247 Ordered Domain-A214gly - 297-526 Ordered Domain;
N-Disordered Domain 140-247 Ordered Domain-203 Y-A214gly - 297-526 Ordered Domain;
N-Disordered Domain 140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain;
N-Disordered Domain 140-247 Ordered DomalnAPMX-203Y-A214gly - 297-526 Ordered Domain;
N-Disordered Domain • 140-247 Ordered DomainAPMX-203Y-A214gly - 297-526 Ordered Domain A297gly;
N-Disordered Domain 140-247 Ordered Domaln-A214gly - 297-526 Ordered Domain-329YA297gly;
N-Disordered Domain 140-247 Ordered Domaln-203Y-A214gly - 297-526 Ordered Domain 329YA297gly;
N-Disordered Domain 140-247 Ordered DomalnAPMX-203Y-A214gly - 297-526 Ordered Domaln-329YA297gly;
N-Dlsordered Domain 140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain;
N-Dlsordered Domain 140-247 Ordered DomalnAPMX - L-Disordered Domain - 297-526 Ordered Domain;

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EP24754502.3A 2023-07-21 2024-07-19 Zusammensetzungen zur abgabe von plasmodium-antigenen und zugehörige verfahren Pending EP4746902A2 (de)

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PCT/US2024/038904 WO2025024335A2 (en)	2023-07-21	2024-07-19	Compositions for delivery of plasmodium antigens and related methods

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