Optimization of human anti-HBsAg scFv secretary expression in Escherichia coli / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 50-52, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-332432
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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Proteins
/
Immunoglobulin Variable Region
/
Hepatitis Antibodies
/
Genetic Engineering
/
Gene Expression
/
Culture Media
/
Protein Transport
/
Allergy and Immunology
/
Escherichia coli
/
Genetics
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2009
Type:
Article
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