HIGH-LEVEL RECOMBINANT PROTEIN-PRODUCTION IN BIOREACTORS USING THE BACULOVIRUS INSECT CELL EXPRESSION SYSTEM

被引:100
作者
CARON, AW [1 ]
ARCHAMBAULT, J [1 ]
MASSIE, B [1 ]
机构
[1] NATL RES COUNCIL CANADA,INST RECH BIOTECHNOL,MONTREAL H4P 2R2,QUEBEC,CANADA
关键词
25;
D O I
10.1002/bit.260361108
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In order to develop an efficient process for large‐scale production of recombinant protein, various factors were studied which affect the productivity of Sf‐9 (Spodoptera frugiperda) insect cells when using the baculovirus expression system. It was shown that upon infection with the Bac‐BRV6L recombinant baculovirus, the level per cell of VP6 (a bovine rotavirus nucleocapsid protein) would drop 10‐fold when host cell density at the time of infection increased from 2 × 106 to 3 × 106 cells/mL. The decrease was found to be totally reversible by culture medium renewal after infection, even when cells were infected at the stationary phase. Recombinant protein production was 4–6 times higher using TNMFH medium supplemented with 10% fetal bovine serum (FBS) than in IPL/41 serum‐free medium. Fine‐tuning of infection parameters in a 4‐L surface‐aerated bioreactor resulted in the production of typically 350 mg/L of VP6 protein, representing more than 25% of total cell proteins. Copyright © 1990 John Wiley & Sons, Inc.
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页码:1133 / 1140
页数:8
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