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Production of a Hepatitis E Vaccine Candidate Using the Pichia pastoris Expression System

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Vaccine Design

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2412))

Abstract

Hepatitis E virus (HEV) is associated with acute hepatitis disease, which may lead to chronic disease in immunocompromised individuals. The disease is particularly severe among pregnant women (20–30% mortality). No vaccine is available to combat the HEV except Hecolin, which is available only in China. Virus-like particle (VLP) generated from the capsid protein (ORF2) of HEV is known to be a potent vaccine antigen against HEV. Hecolin consists of 368–606 amino acid (aa) region of the capsid protein of HEV, which forms a VLP. It is expressed and purified from the inclusion bodies of E. coli. Here, we describe a method to express the 112-608aa region of the capsid protein (ORF2) of genotype-1 HEV in Pichia pastoris (P. pastoris) and purify VLPs from the culture medium. 112-608aa ORF2 VLPs are secreted into the culture medium in a methanol inducible manner. The purified VLPs are glycosylated and induce robust immune response in Balb/c mice. Further, 112-608aa ORF2 VLPs are bigger than the 368–606 VLP present in Hecolin, which may help them in inducing a superior immune response. P. pastoris offers a robust and economical heterologous expression system to produce large quantities of glycosylated 112-608aa ORF2 VLP, which appears to be a promising vaccine candidate against the HEV.

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Correspondence to Milan Surjit .

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Gupta, J., Kumar, A., Surjit, M. (2022). Production of a Hepatitis E Vaccine Candidate Using the Pichia pastoris Expression System. In: Thomas, S. (eds) Vaccine Design. Methods in Molecular Biology, vol 2412. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1892-9_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1892-9_7

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