DK2970876T3 - Serumfrit celledyrkningsmedium - Google Patents

Serumfrit celledyrkningsmedium Download PDF

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DK2970876T3
DK2970876T3 DK14723561.8T DK14723561T DK2970876T3 DK 2970876 T3 DK2970876 T3 DK 2970876T3 DK 14723561 T DK14723561 T DK 14723561T DK 2970876 T3 DK2970876 T3 DK 2970876T3
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cell
protein
ornithine
cell culture
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Amy Johnson
Shawn Lawrence
Shadia Oshodi
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Regeneron Pharma
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Claims (15)

1. Celledyrkningsmedium, der er serumfrit, og som omfatter > 0,09 mM ± 0,014 mM ornithin og > 0,20 ± 0,03 mM putrescin.
2. Celledyrkningsmedium ifølge krav 1, der omfatter 0,3 ± 0,05 mM til 0,9 ± 0,14 mM ornithin.
3. Celledyrkningsmedium ifølge et hvilket som helst af kravene 1 og 2, der omfatter: (a) ornithin ved 0,6 ± 0,09 mM; og/eller (b) putrescin ved 0,714 ± 0,11 mM.
4. Celledyrkningsmedium ifølge et hvilket som helst af de foregående krav, hvor: (a) mediet er hydrolysatfrit eller er kemisk defineret; (b) mediet omfatter >40 + 6 mM af en blanding af aminosyrer eller salte deraf, fortrinsvis hvor blandingen af aminosyrer består af alanin, arginin, asparagin, asparaginsyre, cystein, glutaminsyre, glycin, histidin, isoleucin, leucin, lysin, methionin, phenylalanin, prolin, serin, threonin, tryptophan, tyrosin og valin; (c) mediet omfatter én eller flere fedtsyrer, fortrinsvis hvor den ene eller flere fedtsyrer er udvalgt fra gruppen bestående af linolsyre, linolensyre, thioctsyre, oleinsyre, palmitinsyre, stearinsyre, arachinsyre, arachidonsyre, laurinsyre, behensyre, decansyre, dodecansyre, hexansyre, lignocerinsyre, myristinsyre og octansyre; (d) mediet omfatter en blanding af nukleosider, fortrinsvis hvor blandingen af nukleosider omfatter én eller flere af adenosin, guanosin, cytidin, uridin, thymidin og hypoxanthin; (e) mediet omfatter adenosin, guanosin, cytidin, uridin, thymidin og hypoxanthin; (f) mediet omfatter én eller flere divalente kationer, fortrinsvis hvor den divalente kation er magnesium, calcium, eller begge; og/eller (g) dyrkningsmediet omfatter Ca2+ og Mg2+.
5. Fremgangsmåde til dyrkning af celler, der omfatter følgende trin: (a) tilvejebringelse af et celledyrkningsmedium ifølge et hvilket som helst af de foregående krav, og (b) opformering eller opretholdelse af en celle i celledyrkningsmediet for at danne en cellekultur.
6. Fremgangsmåde ifølge krav 5, hvor: (a) cellen er udvalgt fra gruppen bestående af pattedyrscelle, fjerkræcelle, insektcelle, bakteriecelle og gærcelle, fortrinsvis hvor cellen er en CHO-celle; og/eller (b) cellen udtrykker et protein af interesse.
7. Fremgangsmåde ifølge krav 6, hvor cellen udtrykker et protein af interesse, hvor: (a) proteinet af interesse er et antigenbindingsprotein; (b) proteinet af interesse omfatter et Fc-domæne; og/eller (c) proteinet af interesse er et receptor-Fc-fusionsprotein, fortrinsvis hvor receptor-Fc-fusionsproteinet er et trap-protein.
8. Fremgangsmåde ifølge krav 7, hvor proteinet af interesse er et receptor-Fc-fusionsprotein, der er et trap-protein, hvor trap-proteinet er en IL-1-antagonist eller en VEGF-antagonist.
9. Fremgangsmåde ifølge krav 6, hvor proteinet af interesse er et antistof eller et antistoffragment, fortrinsvis hvor antistoffet eller antistoffragmentet er et rekombinant humant antistof eller fragment deraf.
10. Fremgangsmåde ifølge et hvilket som helst af kravene 5 til 9, hvor: (a) cellerne har en gennemsnitlig fordoblingstid på < 30 timer; (b) cellerne har en gennemsnitlig fordoblingstid på < 24 timer; (c) cellerne har en gennemsnitlig fordoblingstid, der er mindst én tredjedel af den for celler dyrket i et celledyrkningsmedium, der indeholder < 0,3 ± 0,045 mM ornithin og < 0,2 ± 0,03 mM putrescin; (d) cellekulturen er i stand til at opnå en levedygtig celleantalstæthed, der er mindst 15 % større end en tilsvarende cellekultur i medier, der indeholder < 0,09 ± 0,014 mM ornithin og < 0,2 ± 0,03 mM putrescin; (e) cellekulturen er i stand til at opnå en levedygtig celleantalstæthed, der er mindst 3 gange større end en tilsvarende cellekultur i et tilsvarende celledyrkningsmedium, der indeholder < 0,09 ± 0,014 mM ornithin og < 0,2 ± 0,03 mM putrescin; og/eller (f) hvor fremgangsmåden omfatter trinnet med tilsætning af ét eller flere anvendelsesstedstilsætninger til celledyrkningsmediet, fortrinsvis hvor anvendelsesstedstilsætningerne omfatter én eller flere af NaHCO3, glutamin, insulin, glucose, CuSCU, ZnSCU, FeCl3, NiSCU, Na4-EDTA og Na3-citrat, og/eller hver af NaHCC>3, glutamin, insulin, glucose, CuSCU, ZnSCU, FeCl3, NiSCU, Na4-EDTA og Na3-citrat tilsættes mediet som anvendelsesstedstilsætninger.
11. Fremgangsmåde til frembringelse af et protein, hvilken fremgangsmåde omfatter følgende trin: (a) indføring i en celle af en nukleinsyre, der omfatter en sekvens, der koder for et protein af interesse; (b) udvælgelse af en celle, der bærer nukleinsyren; (c) dyrkning af den valgte celle i et celledyrkningsmedium ifølge et hvilket som helst af kravene 1-4 eller ifølge fremgangsmåden ifølge et hvilket som helst af kravene 5-10; og (d) ekspression af proteinet af interesse i cellen, hvor proteinet af interesse sekreteres i mediet.
12. Fremgangsmåde ifølge krav 11, hvor cellen er en CHO-celle, 293-celle eller BHK-celle.
13. Fremgangsmåde ifølge krav 11 eller 12, hvor: (a) proteinet af interesse er et antigenbindingsprotein; (b) proteinet af interesse omfatter et Fc-domæne; og/eller (c) proteinet af interesse er udvalgt fra gruppen bestående af receptor-Fc-fusionsprotein (TRAP), opløseligt TCR-Fc-fusionsprotein, antistof, Fc-fusionsprotein og ScFv-protein.
14. Fremgangsmåde ifølge et hvilket som helst af kravene 11 til 14, hvor: (a) proteinet af interesse frembringes ved en gennemsnitlig 7-dages titer, der er mindst 7 % større end den gennemsnitlige 7-dages titer frembragt af en tilsvarende celle i et celledyrkningsmedium, der indeholder mindre end 0,09 ± 0,014 mM ornithin og mindre end 0,2 ± 0,03 mM putrescin; (b) proteinet af interesse frembringes ved en gennemsnitlig 7-dages titer, der er mindst 14 % større end den gennemsnitlige 7-dages titer frembragt af en tilsvarende celle i et celledyrkningsmedium, der indeholder mindre end 0,09 ± 0,014 mM ornithin og mindre end 0,2 ± 0,03 mM putrescin; (c) proteinet af interesse frembringes ved en gennemsnitlig 7-dages titer, der er mindst 80 % større end den gennemsnitlige 7-dages titer frembragt af en tilsvarende celle i et celledyrkningsmedium, der indeholder mindre end 0,09 ± 0,014 mM ornithin og mindre end 0,2 ± 0,03 mM putrescin; (d) proteinet af interesse frembringes ved en gennemsnitlig 7-dages titer, der er mindst 2-gange større end den gennemsnitlige 7-dages titer frembragt af en tilsvarende celle i et celledyrkningsmedium, der indeholder mindre end 0,09 ± 0,014 mM ornithin og mindre end 0,2 ± 0,03 mM putrescin; og/eller (e) proteinet af interesse frembringes ved en gennemsnitlig 7-dages titer, der er mindst 3-gange større end den gennemsnitlige 7-dages titer frembragt af en tilsvarende celle i et celledyrkningsmedium, der indeholder mindre end 0,09 ± 0,014 mM ornithin og mindre end 0,2 ± 0,03 mM putrescin.
15. Fremgangsmåden ifølge et hvilket som helst af kravene 11 til 14, hvor proteinet af interesse er et rekombinant humant antistof.
DK14723561.8T 2013-03-15 2014-03-14 Serumfrit celledyrkningsmedium DK2970876T3 (da)

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KR (6) KR102166166B1 (da)
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AU2020200052B2 (en) 2023-01-19
MX2021001859A (es) 2021-04-19
HUE040233T2 (hu) 2019-02-28
SG10201707153UA (en) 2017-10-30
CA3238123A1 (en) 2014-09-18
EP3378932A1 (en) 2018-09-26
IL281515A (en) 2021-04-29
SG10202107444XA (en) 2021-08-30
IL289724A (en) 2022-03-01
NZ711935A (en) 2021-04-30
AR095196A1 (es) 2015-09-30
US20220372436A1 (en) 2022-11-24
EP2970876A1 (en) 2016-01-20
JP2022172390A (ja) 2022-11-15
KR20220116571A (ko) 2022-08-23
TWI629359B (zh) 2018-07-11
CA2906768C (en) 2022-08-30
KR20210031778A (ko) 2021-03-22
KR20150128785A (ko) 2015-11-18
EP3919611A1 (en) 2021-12-08
TW201835325A (zh) 2018-10-01
KR102229380B1 (ko) 2021-03-18
IL241195B (en) 2019-05-30
AU2020200052A1 (en) 2020-01-30
CA3238121A1 (en) 2014-09-18
EA202190326A2 (ru) 2021-05-31

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