[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.
Department of Biochemisty and Molecular Biology, Shanxi Medical University, Taiyuan 030001, China.
Jian-Lin
JL
Zhang
Jian-Hua
JH
Wang
Ru-Gang
RG
Zhong
Bo
B
Niu
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