| Title | [Optimization of human anti-HBsAg scFv secretary expression in Escherichia coli] | | Author(s) | Zhang JL, Wang JH, Zhong RG, Niu B | | Institution | Department of Biochemisty and Molecular Biology, Shanxi Medical University, Taiyuan 030001, China. | | Source | Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 2009 Feb; 23(1):50-2. | | 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. | | Language | chi | | Pub Type(s) | English Abstract
Journal Article
| | PubMed ID | 19799018 |
|
