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Vol. 8, Issue 4 (2019)

Production of hepatitis B surface antigen (HBsAg) from Escherichia coli for development of diagnostic kit

Author(s):
Bhavya K, Neha Guleria, Ramanjini Gowda PH and Satish Kumar K
Abstract:
Hepatitis B virus (HBV) infection is a worldwide health problem, which can lead to severe liver disease mainly hepatocellular carcinoma and cirrhosis. The present investigation lays emphasis on expression of HBsAg gene in E. coli. The confirmed recombinant pET28a+HBsAg clone was transformed into E. coli strain BL21 (DE3) PlysS cells. The positive clones were used for protein expression studies and induction parameters viz., IPTG concentration, temperature, induction time and pH of the medium were standardized to produce optimum HBsAg protein yield. The highest protein expression was recorded in clone 2 when it was subjected to 1.5 µM IPTG (384 µg mL-1), pH at 8 (378 µg mL-1) and induction time for 8 h (360 µg mL-1) with 35 °C induction temperature. The cells pelleted was lysed by sonication and protein purification was done using fused His-tag via Ni-NTA agarose-based method. The purified protein from clone 2 was subjected for 12 % SDS-PAGE analysis. The presence of Ì´ 27 kDa protein band confirmed the expression of HBsAg protein in E. coli which was further confirmed through western blot.
Pages: 515-521  |  624 Views  213 Downloads


The Pharma Innovation Journal
How to cite this article:
Bhavya K, Neha Guleria, Ramanjini Gowda PH, Satish Kumar K. Production of hepatitis B surface antigen (HBsAg) from <em>Escherichia coli </em>for development of diagnostic kit. Pharma Innovation 2019;8(4):515-521.

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