DK2907873T3 - Samtidig integreret udvælgelse og udvikling af antistof-/protein-ydelse og deres eksprimering i produktionsværter - Google Patents
Samtidig integreret udvælgelse og udvikling af antistof-/protein-ydelse og deres eksprimering i produktionsværter Download PDFInfo
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- DK2907873T3 DK2907873T3 DK15156820.1T DK15156820T DK2907873T3 DK 2907873 T3 DK2907873 T3 DK 2907873T3 DK 15156820 T DK15156820 T DK 15156820T DK 2907873 T3 DK2907873 T3 DK 2907873T3
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- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B30/00—Methods of screening libraries
- C40B30/04—Methods of screening libraries by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1058—Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
- C40B50/06—Biochemical methods, e.g. using enzymes or whole viable microorganisms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
Claims (15)
1. Fremgangsmåde til udvikling og eksprimering af et antistof i en eukaryotisk celleproduktionsvært; hvilken fremgangsmåde omfatter: a. udvælgelse af et modelantistof; b. udvikling af modelantistoffet for at muliggøre produktion af et sæt mutante antistoffer i en eukaryotisk celleproduktionsvært; c. screening af de mutante antistoffer efter i det mindste én forudbestemt egenskab, karakteristik eller aktivitet; d. udvælgelse af et op-muteret antistof blandt sættet af mutante antistoffer, baseret på optimering af den i det mindste ene forudbestemte egenskab, karakteristik eller aktivitet, i sammenligning med den samme egenskab, karakteristik eller aktivitet for modelantistoffet; og e. eksprimering af det op-muterede antistof i den samme eukaryotiske celleproduktionsvært som i trin (b) i en vilkårlig kommerciel skala.
2. Fremgangsmåde ifølge krav 1, hvor udvælgelsestrinnet (a) omfatter: a1. generering af et anti-antigenantistofbibliotek i en eukaryotisk celleproduktionsvært med antistofcelleoverfladefremvisning; a2. screening af biblioteket efter i det mindste en forudbestemt egenskab, karakteristik eller aktivitet; og a3. udvælgelse af et modelantistof fra biblioteket.
3. Fremgangsmåde ifølge krav 1, hvor den eukaryotiske celleproduktionsvært vælges blandt 3T3 musefibroblastceller; BHK21 syrisk hamsterfibroblastceller; MDCK-hundeepithelceller; Hela humane epithelceller; PtK1 rottekænguruepithelceller; SP2/0 museplasmaceller; og NSO museplasmaceller; HEK 293 humane embryone nyreceller; COS abenyreceller; CHO, CHO-S-kinesiske hamsterovarieceller, R1 museembryoniske celler; E14.1 museembryoniske celler; H1 humane embryoniske celler; H9 humane embryoniske celler; PER C.6, humane embryoniske celler; S. cerivisiae-gærceller; og picchia-gærceller.
4. Fremgangsmåde ifølge krav 3, hvor den eukaryotiske celleproduktsvært er CHO eller HEK293.
5. Fremgangsmåde ifølge krav 1, hvor screeningstrinnet omfatter fluorescens-aktiveret cellesortering (FACS).
6. Fremgangsmåde ifølge krav 1, hvor udviklingstrinnet omfatter produktion af et sæt af mutante antistoffer dannet af modelantistoffet med m komplementaritetsbestemmende regioner (CDR), hvor m er et helt tal mellem 1 og 6, hver af de nævnte CDR omfatter n aminosyrerester, og fremgangsmåden omfatter: a. generering af m x n separate sæt af antistoffer, idet hvert sæt omfatter elementantistoffer med et antal på X forskellige forudbestemte aminosyrerester ved en enkelt forudbestemt position i CDR; hvor hvert sæt af antistoffer afviger i den enkelte forudbestemte position; og antallet af forskellige elementantistoffer, som er genereret, er ækvivalent med m x n x X.
7. Fremgangsmåde ifølge krav 6, hvor m er 6.
8. Fremgangsmåde ifølge krav 1, hvor modelantistoffet er et antistoffragment valgt fra en tung kæde, let kæde, variabelt domæne, konstant domæne, hypervariabel region, komplementaritetsbestemmende region 1 (CDR1), komplementaritetsbestemmende region 2 (CDR2), og komplementaritetsbestemmende region 3 (CDR3).
9. Fremgangsmåde ifølge krav 1, hvor udviklingstrinnet (b) omfatter h. generering af n-1 separate sæt af mutante polypeptider ud fra modelantistoffet, idet hvert sæt omfatter elementpolypeptider med et antal på X forskellige forudbestemte aminosyrerester i en enkelt forudbestemt position af polypeptidet; hvor hvert sæt af polypeptider afviger i den enkelte forudbestemte position; og antallet af forskellige elementpolypeptider, som genereres, er ækvivalent med [n-1] xX; og hvor screeningstrinnet (c) omfatter: i. assaying hvert elementpolypeptid med henblik på i det mindste én forudbestemt egenskab, karakteristik eller aktivitet; j. identifikation af enhver ændring i den nævnte egenskab, karakteristik eller aktivitet for elementpolypeptidet i forhold til modelantistoffet; k. tilvejebringelse af en funktionel afbildning, hvor den funktionelle afbildning anvendes til at identificere positioner og mutationer i det mutante polypeptid, som resulterer i en op-mutant og/eller en stille mutation i sammenligning med modelantistoffet.
10. Fremgangsmåde ifølge krav 9, hvor X er 19.
11. Fremgangsmåde ifølge krav 10, hvor genereringstrinnet omfatter: i. underkastning af et codon-indholdende polynukleotid, som koder for det nævnte modelantistof, for en polymerasebaseret amplifikation under anvendelse af et 64 gange degenereret oligonukleotid for hvert codon, som skal mutageniseres, hvor hvert af de 64 gange degenererede oligonukleotider omfatter en første homolog sekvens og en degenereret Ν,Ν,Ν trippelsekvens, for derved at generere et sæt af polynukleotidafkom; og ii. underkastning af sættet af polynukleotidafkom for klonal amplifikation således, at mutante polypeptider, som kodes for af polynukleotidafkommene, ekspri-meres.
12. Fremgangsmåde ifølge krav 1, hvor den forudbestemte egenskab, karakteristik eller aktivitet vælges fra reduktion af antistofproteinaggregation, forbedring af antistofstabilitet, forøget antistofopløselighed, indføring af glycosylationssteder, indføring af konjugationssteder, reduktion af immunogenicitet, forbedring af proteineksprimering, forøgelse af antigenaffinitet, formindskelse af antigenaffinitet, ændring af bindingsaffinitet, ændring i immunogenicitet, og forbedring af specificitet.
13. Fremgangsmåde ifølge krav 2, hvor anti-antigenantistofbiblioteket er et humaniseret anti-antigenantistofbibliotek.
14. Fremgangsmåde ifølge krav 1, hvor udvælgelsestrinnet (d) omfatter: udvælgelse af op-muteringsantistoffet baseret på forbedret eksprimering, i sammenligning med modelantistoffet; og hvor eksprimeringstrinnet (e) omfatter fremstilling af det op-muterede antistof i den samme eukaryotiske celleproduktionsvært som i udviklingstrinnet (b).
15. Fremgangsmåde ifølge krav 1, hvor udviklingstrinnet (b) omfatter én af comprehensive positional evolution (CPE); comprehensive positional insertion evolution (CPI); comprehensive positional deletion evolution (CPD); comprehensive positional evolution (CPE) efterfulgt af combinatorial protein synthesis (CPS); comprehensive positional deletion evolution (CPD) efterfulgt af combinatorial protein synthesis (CPS); eller comprehensive positional deletion evolution (CPD) efterfulgt af combinatorial protein synthesis (CPS).
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US27116809P | 2009-07-17 | 2009-07-17 | |
| EP10800618.0A EP2454376B1 (en) | 2009-07-17 | 2010-07-16 | Simultaneous, integrated selection and evolution of antibody/protein performance and expression in production hosts |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2907873T3 true DK2907873T3 (da) | 2016-06-13 |
Family
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Family Applications (5)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK19210622.7T DK3636759T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af antistof/protein-ydelse og ekspression hos produktionsværter |
| DK16196481.2T DK3156485T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af antistof/protein-ydelser og eksprimering i produktionsværter |
| DK18183032.4T DK3406717T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af antistof/protein- performance og ekspression hos produktionsværter |
| DK16157965.1T DK3042957T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af human protein-ydelse og ekspression i produktionsværter |
| DK15156820.1T DK2907873T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig integreret udvælgelse og udvikling af antistof-/protein-ydelse og deres eksprimering i produktionsværter |
Family Applications Before (4)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK19210622.7T DK3636759T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af antistof/protein-ydelse og ekspression hos produktionsværter |
| DK16196481.2T DK3156485T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af antistof/protein-ydelser og eksprimering i produktionsværter |
| DK18183032.4T DK3406717T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af antistof/protein- performance og ekspression hos produktionsværter |
| DK16157965.1T DK3042957T3 (da) | 2009-07-17 | 2010-07-16 | Samtidig, integreret udvælgelse og udvikling af human protein-ydelse og ekspression i produktionsværter |
Country Status (19)
| Country | Link |
|---|---|
| US (5) | US8859467B2 (da) |
| EP (6) | EP3636759B1 (da) |
| JP (3) | JP6193570B2 (da) |
| KR (5) | KR20190017070A (da) |
| CN (2) | CN102625848A (da) |
| AU (5) | AU2010273974B2 (da) |
| BR (2) | BR112012001171B1 (da) |
| CA (2) | CA3034484A1 (da) |
| DK (5) | DK3636759T3 (da) |
| ES (3) | ES2701931T3 (da) |
| HK (1) | HK1210623A1 (da) |
| HU (1) | HUE029328T2 (da) |
| IN (1) | IN2012DN00737A (da) |
| MX (2) | MX347654B (da) |
| MY (2) | MY160472A (da) |
| PL (2) | PL3042957T3 (da) |
| PT (1) | PT3042957T (da) |
| SG (3) | SG177687A1 (da) |
| WO (1) | WO2011009058A2 (da) |
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