Optimization of human anti-HBsAg scFv secretary expression in Escherichia coli / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>