A Study on the Optimization of Condition for Soluble Expression of Fusarium-specific scFv Antibody in Escherichia coli

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.034

Abstract

Abstract: The objectives of the work is to optimize the conditions for inducing expression, and obtain the soluble and high-yield expression of a Fusarium-specific single-chain variable fragment(scFv)antibody in the periplasmic space of Escherichia coli XL1-Blue. The recombinant plasmid containing a Fusarium-specific scFv antibody FvSG7 was transferred into E. coli XL1-Blue. After the culture medium for inducing selected, the soluble expression level and activity of FvSG7 antibody were analyzed by Western blot and ELISA detection for studying the influence of temperature, IPTG concentration and induction time on expression level. The maximum productivity of soluble FvSG7 antibody was obtained after induction at 25℃ for 2 h, with a final concentration of 0.1 mmol/L β-D-thiogalactopyranoside(IPTG)and the cultured bacteria growing to the OD₆₀₀ value of 0.5. In conclusion, the soluble expression of FvSG7 antibody in the periplasm of E. coli XL1-Blue increased significantly by optimizing the expression’s conditions of induction temperature, IPTG concentration and induction time.

Key words: single-chain variable fragment antibody, soluble expression, Escherichia coli, optimization

Hu Zuquan, Li Heping, Wu Ping, Liao Yucai, Zhang Jingbo. A Study on the Optimization of Condition for Soluble Expression of Fusarium-specific scFv Antibody in Escherichia coli[J]. Biotechnology Bulletin, 2015, 31(9): 238-243.

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