Optimization of human anti-HBsAg scFv secretary expression in Escherichia coll
10.3760/cma.j.issn.1003-9279.2009.01.018
- VernacularTitle:人源性抗HBsAg单链抗体在大肠埃希菌中分泌表达条件的研究
- Author:
Jian-Lin ZHANG
1
;
Jian-Hua WANG
;
Ru-Gang ZHONG
;
Bo NIU
Author Information
1. 山西医科大学
- Keywords:
Hepatitis B antibodies;
Hepititis B surface antigens;
Secretary expression;
Escherichia coli
- From:
Chinese Journal of Experimental and Clinical Virology
2009;23(1):50-52
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the conditions for high expression of anti-HBsAg scFv A-15 in E. coli, increase the production of the scFv in the culture medium. Methods By changing induction occasion, concentration of inductor IPTG and induction time, influence of various conditions on expression of anti-HBsAg scFv A-15 was analyzed through ELISA. In addition, the effects of sucrose, glycine and Triton X-100 at different concentrations on the scFv excretion into culture medium was evaluation. Results The optimal expression conditions were as follows: the induction was started after culturing for 4 h, the concentration of IPTG was 0.5 mmol/L,and the induction lasted for 8 h .The scFv affinity in culture medium with 0.3 mol/L sucrose,2% glycine, 1% Triton X-100,16.78-fold higher, respectively than that without the three chemicals. The final yield of anti-HBsAg scFv A-15 was estimated to be 7.4 mg/L. Conclusion The conditions for production of anti-HBsAg scFv A-15 were optimized,which provides a practical method for more efficient production of the scFv in E. coli for further studying structure and function.
