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. 2012 Jul 28;27(4):254–258. doi: 10.1007/s12250-012-3256-7

Expression and purification of enterovirus type 71 polyprotein P1 using Pichia pastoris system

Xue Han 1,✉, Xiaoling Ying 1, Hao Huang 1, Shili Zhou 1, Qi Huang 1
PMCID: PMC8218036  PMID: 22899434

Abstract

Enterovirus type 71(EV71) causes severe hand-foot-and-mouth disease (HFMD) resulting in hundreds of deaths of children every year; However, currently, there is no effective treatment for EV71. In this study, the EV71 poly-protein (EV71-P1 protein) gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GS115. The EV71 P1 protein with a molecular weight of 100 kD was produced and secreted into the medium. The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70%. The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits. We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.

Keywords: EV71-P1 protein, Expression, Purification, Immunogenicity, Pichia pastoris

Footnotes

Foundation items: Key Project of Science and Technology Department of Wuhan (zz2011-12); Key Project of the Educational Bureau of Wuhan (sz2011-13-10); Project of Science and Technology Department of Hubei (2010CDB04801); The State Key Laboratory program of Viral Genetic Engineering (2010KF10).

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Articles from Virologica Sinica are provided here courtesy of Wuhan Institute of Virology, Chinese Academy of Sciences

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