Pak. J. Bot., 44: 355-359, Special Issue March 2012. |
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Updated: 26-04-12 | ||||
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EXPRESSION OF PRES2/S ANTIGEN OF HEPATITIS B VIRUS ISOLATED FROM PAKISTAN IN YEAST CELLS
ALI RAZA AWAN1*, MUHAMMAD YASIR ZAHOOR2, MUHAMMAD MOHSIN JAVED3, MASROOR ELLAHI BABAR1AND ZAFAR SALEEM2
Abstract: Recombinant vaccines are an indispensable component of disease management of Hepatitis B infection. Genetic evolution of Hepatitis B Virus (HBV) necessitates the production of new types of antigens for next generation vaccine. This study aimed the expression of HBV middle surface (PreS2/S) protein in Pichia pastoris for production of an antigen for improved HBV vaccine formulation. A PreS2/S antigen-gene was cloned into a vector (pPICZAα) for secretory expression in P. pastoris. Expression of recombinant P. pastoris was optimized under control of AOX1 promoter to produce PreS2/S recombinant protein, which was purified through column chromatography. Specificity of the protein was confirmed through SDS-PAGE, western blot and ELISA. For immunization study, recombinant PreS2/S antigen and commercially available HBV vaccine comprising HBsAg alone were injected to immunize four groups of rabbits. The results of analysis of immunization showed that recombinant PreS2/S protein produced double antibody-titer than that of commercially available HBsAg indicating that PreS2/S antigen could be an ideal candidate for new generation HBV vaccines. This is the first report of secretory expression of biologically active PreS2/S antigen in P. pastoris from Pakistan and it will help to produce more effective vaccine to immunize 180 million people of Pakistan against HBV.
1Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Outfall Road, Lahore Pakistan 2Center of Excellence in Molecular Biology, University of the Punjab, 87 West Canal Bank Road, Lahore 3Institute of Industrial Biotechnology, GC University, Lahore *Corresponding author: arawan77@gmail.com |
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