RENATURATION, PURIFICATION AND CHARACTERIZATION OF RECOMBINANT FAB-FRAGMENTS PRODUCED IN ESCHERICHIA-COLI

被引：403
作者：
BUCHNER, J
RUDOLPH, R
机构：
[1] BOEHRINGER MANNHEIM GMBH,BIOCHEM RES CTR,NONNENWALD 2,W-8122 PENZBERG,GERMANY
[2] UNIV REGENSBURG,INST BIOPHYS & PHYS BIOCHEM,W-8400 REGENSBURG,GERMANY
来源：
BIO-TECHNOLOGY | 1991年 / 9卷 / 02期
关键词：
D O I：
10.1038/nbt0291-157
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：
Cytoplasmatic expression of murine antibody chains in Escherichia coli results in the formation of insoluble and inactive protein aggregates (inclusion bodies). By systematic variation of the parameters influencing the folding, formation of disulfide bonds and association of the constituent polypeptide chains, we have designed a renaturation procedure allowing the production of microbially expressed F(ab)-fragments at yields up to 40 percent of the total amount of recombinant protein. The strategy of optimization is generally applicable for disulfide containing proteins produced as inclusion bodies in bacteria. The purified recombinant antibody fragments obtained are identical with the native murine F(ab) in all functional and physicochemical parameters tested.
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页码：157 / 162
页数：6
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