Design and production of a multiepitope construct derived from hepatitis E virus capsid protein.

Journal of Medical Virology
Reza TaherkhaniManoochehr Makvandi

Abstract

The aim of this study was to design a high density multiepitope protein, which can be a promising multiepitope vaccine candidate against Hepatitis E virus (HEV). Initially, conserved and antigenic helper T-lymphocyte (HTL) epitopes in the HEV capsid protein were predicted by in silico analysis. Subsequently, a multiepitope comprising four HTL epitopes with high-affinity binding to the HLA molecules was designed, and repeated four times as high density multiepitope construct. This construct was synthesized and cloned into pET-30a (+) vector. Then, it was transformed and expressed in Escherichia coli BL21 cells. The high density multiepitope protein was purified by Ni-NTA agarose and concentrated using Amicon filters. Finally, the immunological properties of this high density multiepitope protein were evaluated in vitro. The results showed that the high density multiepitope construct was successfully expressed and purified. SDS-PAGE and Western blot analyses showed the presence of a high density multiepitope protein band of approximately 33 kDa. Approximately 1 mg of the purified protein was obtained from each liter of the culture media. Moreover, the purified multiepitope protein was capable of induction of proliferation respons...Continue Reading

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Citations

Jul 23, 2015·Hepatitis Research and Treatment·Fatemeh FarshadpourManoochehr Makvandi
Jun 15, 2016·World Journal of Gastroenterology : WJG·Reza Taherkhani, Fatemeh Farshadpour
Jul 27, 2021·The International Journal of Artificial Organs·Mahbube OujiFatemeh Farshadpour
Sep 25, 2021·Critical Reviews in Microbiology·Roghayeh MohammadzadehHadi Farsiani

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Datasets Mentioned

BETA
Q68985

Methods Mentioned

BETA
ELISA
density gradient centrifugation
Assay
FCS
dissection

Software Mentioned

Ed
Sci
GenScript Rare Codon Analysis Tool
MEGA
Clone Manager Basic
IEDB Analysis Resource
Vector NTI
SYFPEITHI
SPSS

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