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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.
- Source :
-
World Journal of Microbiology & Biotechnology . Dec2014, Vol. 30 Issue 12, p3221-3227. 7p. - Publication Year :
- 2014
-
Abstract
- 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. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09593993
- Volume :
- 30
- Issue :
- 12
- Database :
- Academic Search Index
- Journal :
- World Journal of Microbiology & Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 99075153
- Full Text :
- https://doi.org/10.1007/s11274-014-1749-2