Recombinant protein production using the Semliki Forest Virus expression system

被引：34

作者：

Blasey, HD ^{[1
]}

Lundstrom, K ^{[1
]}

Tate, S ^{[1
]}

Bernard, AR ^{[1
]}

机构：

[1] GLAXO WELLCOME RES & DEV LTD,MED RES CTR,STEVENAGE SG1 2NY,HERTS,ENGLAND

来源：

CYTOTECHNOLOGY | 1997年 / 24卷 / 01期

关键词：

human cyclooxygenase-2; infection optimisation; Semliki Forest Virus; suspension process; transient expression;

D O I：

10.1023/A:1007974121182

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：

We report here the successful scale up of transient recombinant protein expression to litre scale using Semliki Forest Virus System. The expression of bacterial beta-galactosidase was initially compared in BHK and CHO cells and the conditions for optimal infection of BHK cells were identified. 10% FCS in a medium at pH 6.9 and infection in small volumes were found to be optimal. A high MOI results in an increased recombinant protein yield. Stirring does not affect the infection process. Finally we applied these optimal conditions to the production of a microsomal enzyme, human cyclooxygenase-2 in suspension spinners. Five independant productions at the 1 litre scale yielded reproducible substantial amounts of recombinant protein (16 mg microsomal protein 10(9) cells(-1)) with an average specific activity of 3942 +/- 765 pg PGE(2) mu g(-1) microsomal protein 5 min(-1).

引用

页码：65 / 72

页数：8

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