DK2804875T3 - Værktøjer og fremgangsmåder til ekspression af membranproteiner - Google Patents

Værktøjer og fremgangsmåder til ekspression af membranproteiner Download PDF

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DK2804875T3
DK2804875T3 DK13700751.4T DK13700751T DK2804875T3 DK 2804875 T3 DK2804875 T3 DK 2804875T3 DK 13700751 T DK13700751 T DK 13700751T DK 2804875 T3 DK2804875 T3 DK 2804875T3
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protein
host cell
membrane protein
membrane
nanobody
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DK13700751.4T
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Jan Steyaert
Nico Callewaert
Toon Laeremans
Els Pardon
Kristof Vandewalle
Katrien Claes
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Vib Vzw
Univ Brussel Vrije
Univ Gent
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • 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
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • 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
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • C07K14/7158—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for chemokines
    • 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
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
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    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2866—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for cytokines, lymphokines, interferons
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G01N33/5041—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects involving analysis of members of signalling pathways
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
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    • C07—ORGANIC CHEMISTRY
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    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
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    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/715—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons
    • G01N2333/7158—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons for chemokines
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/02—Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
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    • G01—MEASURING; TESTING
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    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/10—Screening for compounds of potential therapeutic value involving cells
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    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/20—Screening for compounds of potential therapeutic value cell-free systems

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Claims (15)

1. Værtscelle, der omfatter en første exogen nukleinsyresekvens, der koder for et membranprotein, og en anden exogen nukleinsyre, der koder for et nanobody, der binder specifikt til en extracellulær eller en intracellulær konformationel epitop på membranproteinet, som hver er under kontrol af en promotor.
2. Værtscelle ifølge krav 1, hvor promotoren er en konstitutiv promotor eller en inducerbar promotor.
3. Værtscelle ifølge et hvilket som helst af kravene 1 eller 2, hvor membranprotein og nanobody udtrykkes sammen.
4. Værtscelle ifølge et hvilket som helst af kravene 1 til 3, hvor membranprotein og/eller nanobody er operabelt koblet til en eller flere subcellulære målretningssekvenser, såsom et ER- eller Golgi-lokaliseringssignal eller et sekretionssignal eller kombinationer deraf.
5. Værtscelle ifølge et hvilket som helst af kravene 1 til 4, hvor membranproteinet er et membranreceptorprotein, såsom en GPCR, eller et membrantransportprotein, såsom en iontransportør.
6. Værtscelle ifølge et hvilket som helst af kravene 1 til 5, hvor nanobody'et stabiliserer membranproteinet i en funktionel konformationel tilstand, såsom en aktiv eller en inaktiv tilstand.
7. Værtscelle ifølge et hvilket som helst af kravene 1 til 6, hvor værtscellen er en eukaryot værtscelle, såsom en gærcelle, en mammaliacelle, en insektcelle, en trådformet svampecelle.
8. Værtscelle ifølge krav 7, hvor gæren er en Pichia-stamme, såsom en Pichia pastoris, eller en Komagataella-stamme, såsom Komagataella pastoris, eller en Hansenula-stamme, såsom Hansenula polymorpha, eller en Yarrowia-stamme, såsom Yarrowia lipolytica, eller en Saccharomyces-stamme, såsom Saccharomyces cerevisiae, og hvor den trådformede svamp er en Aspergillus-stamme, såsom Aspergillus niger eller Aspergillus nidulans, eller en Penicillium-stamme, såsom Penicillium citrinum eller Penicillium chrysogenum, eller en Hypocrea-stamme, såsom Hypocrea jecorina.
9. Værtscelle ifølge et hvilket som helst af kravene 1 til 8, hvor værtscellen er en glycomodificeret værtscelle.
10. Cellekultur af en værtscelle ifølge et hvilket som helst af kravene 1 til 9.
11. Membranpræparat af en værtscelle ifølge et hvilket som helst af kravene 1 til 9 eller af en cellekultur ifølge krav 10 .
12. Fremgangsmåde til forøgelse af produktionen af et membranprotein i en værtscelle, hvilken fremgangsmåde omfatter trinene: a. tilvejebringelse af en værtscelle ifølge et hvilket som helst af kravene 1 til 9, b. dyrkning af værtscellen under betingelser, der er egnet til samtidig ekspression af membranproteinet og nanobody'et, der er rettet mod membranproteinet.
13. Fremgangsmåde til forøgelse af produktionen af et membranprotein i en funktionel konformation i en værtscelle, hvilken fremgangsmåde omfatter trinene: a. tilvejebringelse af en værtscelle ifølge et hvilket som helst af kravene 1 til 9, b. dyrkning af værtscellen under betingelser, der er egnet til samtidig ekspression af membranproteinet og nanobody'et, der er rettet mod membranproteinet, i nærvær af en ligand, såsom en agonist, en antagonist, en invers agonist.
14. Fremgangsmåde ifølge krav 12 eller 13, der yderligere omfatter trinet isolering af membranproteinet, nanobody'et eller membranproteinet i kompleks med nanobody'et.
15. Anvendelse af værtscellen ifølge et hvilket som helst af kravene 1 til 9 eller cellekulturen ifølge krav 10 eller membranpræparatet ifølge krav 11 til ligandscreening, lægemiddelscreening, proteinindfangning og oprensning, immunisering, biofysiske studier.
DK13700751.4T 2012-01-19 2013-01-21 Værktøjer og fremgangsmåder til ekspression af membranproteiner DK2804875T3 (da)

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US201261588523P 2012-01-19 2012-01-19
EP12151814.6A EP2617732A1 (en) 2012-01-19 2012-01-19 Tools and methods for expression of membrane proteins
PCT/EP2013/051041 WO2013107905A1 (en) 2012-01-19 2013-01-21 Tools and methods for expression of membrane proteins

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US (3) US9890217B2 (da)
EP (2) EP2617732A1 (da)
AU (1) AU2013211001B2 (da)
CA (1) CA2861393C (da)
DK (1) DK2804875T3 (da)
PT (1) PT2804875T (da)
WO (1) WO2013107905A1 (da)

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* Cited by examiner, † Cited by third party
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AU2008228085B2 (en) 2007-03-22 2011-12-08 Nxera Pharma Uk Limited Mutant G-protein coupled receptors and methods for selecting them
GB0724860D0 (en) 2007-12-20 2008-01-30 Heptares Therapeutics Ltd Screening
DK2742066T3 (da) 2011-08-10 2018-03-05 Heptares Therapeutics Ltd Stabile proteiner
EP2617732A1 (en) 2012-01-19 2013-07-24 Vib Vzw Tools and methods for expression of membrane proteins
ES2676827T3 (es) * 2012-06-01 2018-07-25 Heptares Therapeutics Limited Ensayos
KR102564610B1 (ko) 2015-07-09 2023-08-07 브이아이비 브이지더블유 N- 및 o-글리코실화 패턴이 변이된 당단백질을 생성하는 세포 및 그의 방법 및 용도
IL303865A (en) 2020-12-23 2023-08-01 Regeneron Pharma Methods for obtaining antibodies that bind transmembrane proteins and cells that produce them

Family Cites Families (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0549062B1 (en) 1991-12-23 1999-03-17 Gist-Brocades N.V. A eukaryotic expression system
US5721121A (en) 1995-06-06 1998-02-24 Genentech, Inc. Mammalian cell culture process for producing a tumor necrosis factor receptor immunoglobulin chimeric protein
EP1051493A2 (en) 1998-01-26 2000-11-15 Unilever Plc Method for producing antibody fragments
US6730499B1 (en) 1998-07-03 2004-05-04 Research Corporation Technologies, Inc. Promoter for the Pichia pastoris formaldehyde dehydrogenase gene FLD1
DE60013767T3 (de) 1999-01-19 2009-07-09 Unilever N.V. Verfahren zur herstellung von antikörperfragmenten
WO2001090190A2 (en) 2000-05-26 2001-11-29 National Research Council Of Canada Single-domain antigen-binding antibody fragments derived from llama antibodies
KR100787073B1 (ko) 2000-06-28 2007-12-21 글리코파이, 인크. 변형된 당단백질의 제조방법
DE10062302A1 (de) 2000-12-14 2002-07-11 Aventis Res & Tech Gmbh & Co Sekretionssignalpeptide, deren DNA-Sequenzen, damit herstellbare Expressionsvektoren für eukaryotische Zellen und deren Verwendung zur biotechnologischen Herstellung von Proteinen
GB0110029D0 (en) 2001-04-24 2001-06-13 Grosveld Frank Transgenic animal
WO2003025020A1 (en) 2001-09-13 2003-03-27 Institute For Antibodies Co., Ltd. Method of constructing camel antibody library
JP2005289809A (ja) 2001-10-24 2005-10-20 Vlaams Interuniversitair Inst Voor Biotechnologie Vzw (Vib Vzw) 突然変異重鎖抗体
ES2356307T3 (es) 2002-04-19 2011-04-06 Zymogenetics, L.L.C. Procedimientos para detectar o modular la interacción de un receptor de citocina con su ligando.
GB0228210D0 (en) 2002-12-03 2003-01-08 Babraham Inst Single chain antibodies
JP2007500016A (ja) * 2003-06-11 2007-01-11 バイオジェン・アイデック・エムエイ・インコーポレイテッド 培養においてタンパク質生成を増大させるための方法
WO2006119497A2 (en) 2005-05-04 2006-11-09 University Of Utah Research Foundation Acetylycholine gated ion channel chaperons and methods of using the same
US8815580B2 (en) 2008-08-08 2014-08-26 Vib Vzw Cells producing glycoproteins having altered glycosylation patterns and method and use thereof
CA2749558C (en) 2008-12-08 2019-01-15 Complix Nv Single-chain antiparallel coiled coil proteins
US8440456B2 (en) 2009-05-22 2013-05-14 Vib, Vzw Nucleic acids of Pichia pastoris and use thereof for recombinant production of proteins
US20130137140A1 (en) 2010-06-17 2013-05-30 Universiteit Gent Increased protein expression through increased membrane formation
EP2617732A1 (en) 2012-01-19 2013-07-24 Vib Vzw Tools and methods for expression of membrane proteins

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CA2861393A1 (en) 2013-07-25
EP2804875A1 (en) 2014-11-26
US20230071321A1 (en) 2023-03-09
EP2617732A1 (en) 2013-07-24
PT2804875T (pt) 2017-07-13
EP2804875B1 (en) 2017-04-19
US11479791B2 (en) 2022-10-25
AU2013211001B2 (en) 2017-03-09
WO2013107905A1 (en) 2013-07-25
CA2861393C (en) 2020-02-04
US20180237530A1 (en) 2018-08-23
US20140357521A1 (en) 2014-12-04
AU2013211001A1 (en) 2014-09-04
US9890217B2 (en) 2018-02-13

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