Co-expression of Dsb proteins enables soluble expression of a single-chain variable fragment (scFv) against human type 1 insulin-like growth factor receptor (IGF-1R) in E. coli
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- 作者：Xue-Wen Sun (1)
Xiao-Hua Wang (1)
Yan-Bing Yao (1) - 关键词：Dsb ; E. coli ; IFG ; 1R ; ScFv ; Sulfide bond
- 刊名：World Journal of Microbiology and Biotechnology
- 出版年：2014
- 出版时间：December 2014
- 年：2014
- 卷：30
- 期：12
- 页码：3221-3227
- 全文大小：403 KB
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18. Maskos K, Huber-Wunderlich M, Glockshuber R (2003) D - 作者单位：Xue-Wen Sun (1)
Xiao-Hua Wang (1)
Yan-Bing Yao (1)
1. Department of Medical Technology, Medical College, Hebei University of Engineering, 199 South GuangMing Road, Handan, 056038, China - ISSN：1573-0972
文摘
Type 1 insulin-like growth factor receptor (IGF-1R) is a promising therapeutic target for cancer treatment. A single-chain variable fragment (scFv) against human IGF-1R forms inclusion body when expressed in periplasmic space of E. coli routinely. Here, we described that co-expression of appropriate disulfide bonds (Dsb) proteins known to catalyze the formation and isomerization of Dsb can markedly recover the soluble expression of target scFv in E. coli. A 50?% recovery in solubility of the scFv was observed upon co-expression of DsbC alone, and a maximum solubility (80?%) was obtained when DsbA and DsbC were co-expressed in combination. Furthermore, the soluble scFv present full antigen-binding activity with IGF-1R, suggesting its correct folding. This study also suggested that the selection of Dsb proteins should be tested case-by-case if the approach of co-expression of Dsb system is adopted to address the problem of insoluble expression of proteins carrying Dsb.