[Optimization of human anti-HBsAg scFv secretary expression in Escherichia coli].
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.


CONCLUSIONS
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.
Zhang
Jian-Lin
JL
Department of Biochemisty and Molecular Biology, Shanxi Medical University, Taiyuan 030001, China.
Wang
Jian-Hua
JH
Zhong
Ru-Gang
RG
Niu
Bo
B
D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D006508:Hepatitis Antibodies / Q000235:genetics / Q000378:metabolism*; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D006508:Hepatitis Antibodies; D006514:Hepatitis B Surface Antigens; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins