High-level recombinant protein production in bioreactors using the baculovirus–insect cell expression system

✍ Scribed by Antoine W. Caron; Jean Archambault; Bernard Massie

• Publisher: John Wiley and Sons
• Year: 1990
• Tongue: English
• Weight: 864 KB
• Volume: 36
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.260361108

✦ Synopsis

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 x lo6 to 3 x lo6 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. level^,'^^'^ very little is known pertaining to infection of Sf-9 cells with recombinant baculoviruses.