DK2738259T1 - ADAM6 mus - Google Patents
ADAM6 mus Download PDFInfo
- Publication number
- DK2738259T1 DK2738259T1 DK14154967.5T DK14154967T DK2738259T1 DK 2738259 T1 DK2738259 T1 DK 2738259T1 DK 14154967 T DK14154967 T DK 14154967T DK 2738259 T1 DK2738259 T1 DK 2738259T1
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- Denmark
- Prior art keywords
- mouse
- human
- segments
- variable region
- antigen
- Prior art date
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- 239000000427 antigen Substances 0.000 claims 9
- 102000036639 antigens Human genes 0.000 claims 9
- 108091007433 antigens Proteins 0.000 claims 9
- 210000004027 cell Anatomy 0.000 claims 9
- 241001529936 Murinae Species 0.000 claims 8
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 claims 6
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 claims 6
- 150000007523 nucleic acids Chemical group 0.000 claims 6
- 108091028043 Nucleic acid sequence Proteins 0.000 claims 5
- 238000000034 method Methods 0.000 claims 5
- 102000004169 proteins and genes Human genes 0.000 claims 5
- 108090000623 proteins and genes Proteins 0.000 claims 5
- 210000000349 chromosome Anatomy 0.000 claims 4
- 230000003053 immunization Effects 0.000 claims 3
- 108020004414 DNA Proteins 0.000 claims 2
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 claims 2
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 claims 2
- 238000012258 culturing Methods 0.000 claims 2
- 101000998953 Homo sapiens Immunoglobulin heavy variable 1-2 Proteins 0.000 claims 1
- 108060003951 Immunoglobulin Proteins 0.000 claims 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 claims 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 claims 1
- 102100036887 Immunoglobulin heavy variable 1-2 Human genes 0.000 claims 1
- 210000003719 b-lymphocyte Anatomy 0.000 claims 1
- 239000012634 fragment Substances 0.000 claims 1
- 102000053391 human F Human genes 0.000 claims 1
- 108700031895 human F Proteins 0.000 claims 1
- 210000004408 hybridoma Anatomy 0.000 claims 1
- 238000002649 immunization Methods 0.000 claims 1
- 102000018358 immunoglobulin Human genes 0.000 claims 1
- 210000004988 splenocyte Anatomy 0.000 claims 1
- 210000001519 tissue Anatomy 0.000 claims 1
- 238000011144 upstream manufacturing Methods 0.000 claims 1
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- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
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Claims (18)
1. Mus, som udtrykker antistoffer, hvor hver tung kæde omfatter en human variabel region og en murin konstant region, hvor musen også udtrykker: (i) et murint ADAM6a protein eller ortolog eller homolog heraf, som er funktionelt i en hanmus, og (ii) et murint ADAM6b protein eller ortolog eller homolog heraf, som er funktionelt i en hanmus, og hvori ADAM6a- og ADAM6b-proteinerne eller ortologerne eller homologerne heraf er udtrykt fra en ectopisk nucleinsyresekvens.
2. Mus ifølge krav 1, hvor antistofferne produceret af musen mangler en murin variabel region.
3. Mus ifølge krav 1, hvor den ectopiske nucleinsyresekvens er til stede ved en endogen immunoglobulinlocus.
4. Mus ifølge krav 1, hvor musen har et genom omfattende en deletering af en eller flere murine immunoglobulinvariable regionssegmenter, og insertering af en eller flere humane immunoglobulinvariable regionssegmenter
5. Mus ifølge krav 4, hvor muse DH og JH segmenterne og mindst 3, mindst 10, mindst 20, mindst 40, mindst 60 eller mindst 80 VH segmenter af immu-noglobulinets tunge kædes locus er erstattet med humane immunoglobulinvariable regionssegmenter.
6. Mus ifølge krav 4, hvor alle mus DH, Jh og VH segmenter af immunoglobu-linets tunge kædes locus er erstattet med humant immunoglobulins variable regionssegmenter.
7. Mus ifølge krav 6, hvor nævnte ectopiske nucleinsyresekvens er til stede (a) umiddelbart ved siden af 3’-enden af de humane VH segmenter, (b) mellem to humane VH segmenter, eller (c) mellem to VH segmenter inden for ca. 20 kb til ca. 40 kb af 3’-terminalen af de inserterede humane VH segmenter.
8. Mus ifølge krav 6, hvor erstatningen indbefatter humane VH segmenter VH1 -2 og Vh6-1 , og nævnte ectopiske nucleinsyresekvenser er stede nedstrøms for Vh1-2 segmentet og opstrøms for VH6-1 segmentet.
9. Mus ifølge krav 4, hvor den ectopiske nucleinsyresekvens er tilstødende de humane immunoglobulinvariable regionssegmenter.
10. Mus ifølge krav 8, hvor den ectopiske nucleinsyresekvens er på det samme chromosom som de humane immunglobulinvariable regionssegmenter.
11. Mus ifølge krav 10, hvor chromosomet er et chromosom, som er fundet i en mus af vildtypen, og chromosomet omfatter et humaniseret immunoglobulins tunge kædes variable regions locus.
12. Isoleret celle eller isoleret væv fra en mus ifølge et vilkårligt af kravene 1 -11.
13. Anvendelse af en mus ifølge et vilkårligt af kravene 1 -11 til generering af (i) et kimærisk antistof, som omfatter en human variabel region og en murin konstant region, (ii) et fuldstændigt humant antistof, (iii) et fuldstændigt humant Fab fragment, og/eller (iv) et fuldstændigt humant F(ab) 2 fragment.
14. Fremgangsmåde til generering af et kimærisk antistof specifikt mod et antigen omfattende trinnene: (a) immunisering af en mus ifølge et vilkårligt af kravene 1-11 med antigenet, (b) isolering af mindst en celle fra musen, der producerer et kimærisk antistof specifikt mod antigenet, hvilket kimæriske antistof omfatter en human variabel region, og en murin konstant region, og (c) dyrkning af mindst en celle, der producerer det kimæriske antistof i trin (b) og opnåelse af nævnte antistof, fortrinsvis hvor dyrkningen i trin (c) udføres på mindst en hybridomcelle genereret fra den i det mindste ene celle opnået i trin (b).
15. Fremgangsmåde til generering af et fuldstændigt humant antistof specifikt imod et antigen omfattende trinnene: a) immunisering af en mus ifølge et vilkårligt af kravene 1-11 med antigenet, (b) isolering af mindst en celle fra musen, der producerer et kimærisk antistof specifikt mod antigenet, hvilket kimæriske antistof omfatter en human variabel region, og en murin konstant region, og (c) generering af mindst en celle, som producerer et fuldstændigt humant antistof stammende fra det kimæriske antistof i trin (b) og specifikt mod antigenet, og (d) dyrkning af mindst en celle, der producerer det fuldstændigt humane antistof i trin (c) og opnåelse af nævnte fuldstændigt humane antistof.
16. Fremgangsmåde ifølge et vilkårligt af kravene 14 og 15, hvor den i det mindste ene celle opnået i trin (b) er en splenocyt eller en B-celle.
17. Fremgangsmåde ifølge et vilkårligt af kravene 14 - 16, hvor antistoffet er et monoklonalt antistof.
18. Fremgangsmåde ifølge et vilkårligt af kravene 14-17, hvor immunisering med antigenet i trin (a) udføres med protein, DNA, en kombination af DNA og protein eller celler, der udtrykker antigenet.
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