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Virologica Sinica, 29 (6) : 403-409, 2014
Research Article
Generation of Hepatitis B virus PreS2-S antigen in Hansenula polymorpha
1. State Key Laboratory of Nature Medicines, China Pharmaceutical University, Nanjing 210009, China

2. Institute of Vaccine Research, Jiangsu Simcere Pharmaceutical Group, Nanjing 210042, China
 Correspondence: jianqiangli2003@sina.com;13913925346@126.com
Despite the long availability of a traditional prophylactic vaccine containing the HBV surface antigen (HBsAg) and aluminum adjuvant, nearly 10% of the population remains unable to generate an effective immune response. Previous studies have indicated that hepatitis B virus (HBV) PreS2-S is abundant in T/B cell epitopes, which induces a stronger immune response than HBsAg, particularly in terms of cytotoxic T lymphocyte (CTL) reaction. In the current study, the HBV PreS2-S gene encoding an extra 26 amino acids (PreS2 C-terminus) located at the N-terminus of HBsAg was cloned into the pVCH1300 expression vector. PreS2-S expressed in the methylotrophic yeast, Hansenula polymorpha, was produced at a yield of up to 250 mg/L. Subsequent purifi cation steps involved hydrophobic adsorption to colloidal silica, ion-exchange chromatography and density ultracentrifugation. The fi nal product was obtained with a total yield of ~15% and purity of ~99%. In keeping with previous studies, ~22 nm viruslike particles were detected using electron microscopy. The generated PreS2-S antigen will be further studied for effi cacy and safty in animals.
Received: 24 Dec 2014  Accepted: 27 Sep 2014  Published online: 1 Dec 2014
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