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DK2578688T1 - ADAM6-mus - Google Patents

ADAM6-mus Download PDF

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Publication number
DK2578688T1
DK2578688T1 DK12192727.1T DK12192727T DK2578688T1 DK 2578688 T1 DK2578688 T1 DK 2578688T1 DK 12192727 T DK12192727 T DK 12192727T DK 2578688 T1 DK2578688 T1 DK 2578688T1
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Denmark
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mouse
human
gene segments
gene
antigen
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DK12192727.1T
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DK2578688T3 (da
DK2578688T4 (da
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Lynn Macdonald
Sean Stevens
Andrew J Murphy
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Regeneron Pharma
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Claims (15)

1. Mus, som udtrykker en human immunoglobulins tunge kædes variable region eller et funktionelt fragment heraf fra en modificeret endogen mus’ immunoglobulins tunge kædes locus, hvor musen omfatter en ADAM6 aktivitet, som er funktionel i musen, hvor ADAM6 aktiviteten er kodet af en ectopisk nu-cleinsyresekvens.
2. Mus ifølge krav 1, hvor ADAM6 aktiviteten er kodet af en ADAM6 sekvens flankeret opstrøms, nedstrøms eller opstrøms og nedstrøms af en nucleinsyre-sekvens, som koder for et immunoglobulins variable gensegment.
3. Mus ifølge krav 1 eller 2, hvor musen er en hanmus, eller hvor musen er en hunmus.
4. Mus ifølge et vilkårligt af kravene 1 - 3, hvor den modificerede endogene mus immunoglobulins tunge kædes locus omfatter mindst 18 humane VH gensegmenter, 27 humane DH gensegmenter og seks humane Jh gensegmenter operativt forbundet til en mus’ konstante regionsgen, eller mindst 39 humane VH gensegmenter, 27 humane DH gensegmenter og seks humane Jh gensegmenter operativt forbundet til en mus’ konstante regionsgen, eller mindst 80 humane VH gensegmenter, 27 humane DH gensegmenter og seks humane JH gensegmenter operativt forbundet til en mus’ konstante regionsgen.
5. Mus ifølge et vilkårligt af kravene 1 - 4, hvor musens konstante regionsgen omfatter en sekvens valgt fra et Ch1, et hængsel, et Ch2, et Ch3, et Ch4 og en kombination heraf.
6. Mus ifølge et vilkårligt af kravene 1 - 5, hvor den modificerede endogene mus’ immunoglobulins tunge kædes locus omfatter en deletering af alle murine VH, -Dh og Jh gensegmenter.
7. Mus ifølge et vilkårligt af kravene 1 - 6, hvor den ectopiske nucleinsyrese-kvens koder for et murint ADAM6a protein og/eller ADAM6b protein eller en or-tolog, homolog eller et funktionelt fragment heraf.
8. Fremgangsmåde til modifikation af et immunoglobulins tunge kædes locus fra en mus omfattende: (a) fremstilling af en første modifikation af musens immunoglobulins tunge kæde locus, hvor den første modifikation omfatter placeringen af en eller flere humane immunoglobulingensekvenser og/eller erstatningen af en eller flere sekvenser i museimmunoglobulinets tunge kædes locus, med en eller flere humane immunoglobulingensekvenser, og (b) fremstilling af en anden modifikation af musen, som opretholder ADAM6 aktivitet i musen, fortrinsvis hvor den første og den anden modifikation er fremstillet samtidigt.
9. Fremgangsmåde ifølge krav 8, hvor den første modifikation omfatter erstatningen af et eller flere endogene VH gensegmenter med et eller flere humane VH gensegmenter, fortrinsvis hvor den første modifikation omfatter erstatningen af en endogen tung kædes variable gensekvens med en human tung kædes variable gensekvens.
10. Isoleret celle eller isoleret væv fra en mus ifølge et vilkårligt af kravene 1 -7.
11. Fremgangsmåde til generering af et revers/kimærisk murint/humant antistof specifikt mod et antigen omfattende trinnene: a) immunisering af en mus ifølge et vilkårligt af kravene 1 - 7 med antigenet, b) isolering af mindst en celle fra musen, som producerer et revers/kimæ-risk murint/humant antistof specifikt mod antigenet, og c) dyrkning af mindst en celle, som fremstiller et antistof ifølge trin b) og opnåelse af nævnte antistof, hvor fortrinsvis dyrkningen i trin c) udføres på mindst en hybridomcelle genereret fra den mindst ene celle opnået i trin b).
12. Fremgangsmåde til generering af et fuldt humant antistof specifikt over for et antigen omfattende trinnene: a) immunisering af en mus ifølge et vilkårligt af kravene 1 - 7 med antigenet, b) isolering af den i det mindste ene celle fra musen, som producerer et re-vers/kimærisk murint/humant antistof specifikt mod antigenet, og c) generering af den i det mindste ene celle, som fremstiller et fuldt humant antistof stammende fra antistoffet i trin b) og specifikt mod antigenet, og d) dyrkning af den i det mindste ene celle, som fremstiller et antistof ifølge trin c) og opnåelse af nævnte antistof.
13. Fremgangsmåde ifølge et vilkårligt af kravene 11 og 12, hvor den i det mindste ene celle opnået i trin b) er en splenocyt eller en B-celle.
14. Fremgangsmåde ifølge et vilkårligt af kravene 11 - 13, hvor antistoffet er et monoklonalt antistof.
15. Fremgangsmåde ifølge et vilkårligt af kravene 11 - 14, hvor immuniseringen med antigenet i trin (a) udføres med protein, DNA, en kombination af DNA og protein eller celler, som udtrykker antigenet.
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