Abstract
Hepatitis E virus (HEV) is responsible for hepatitis E, which represents a global public health problem. HEV genotypes 3 and 4 are reported to be zoonotic, and animals are monitored for HEV infection in the interests of public hygiene and food safety. The development of novel diagnostic methods and vaccines for HEV in humans is thus important topics of research. Opening reading frame (ORF) 2 of HEV includes both linear and conformational epitopes and is regarded as the primary candidate for vaccines and diagnostic tests. We investigated the precise location of the HEV epitopes in the ORF2 protein. We prepared four monoclonal antibodies (mAbs) against genotype 4 ORF2 protein and identified two linear epitopes, G438IVIPHD444 and Y457DNQH461, corresponding to two of these mAbs using phage display biopanning technology. Both these epitopes were speculated to be universal to genotypes 1, 2, 3, 4, and avian HEVs. We also used two 12-mer fragments of ORF2 protein including these two epitopes to develop a peptide-based enzyme-linked immunosorbent assay (ELISA) to detect HEV in serum. This assay demonstrated good specificity but low sensitivity compared with the commercial method, indicating that these two epitopes could serve as potential candidate targets for diagnosis. Overall, these results further our understanding of the epitope distribution of HEV ORF2, and provide important information for the development of peptide-based immunodiagnostic tests to detect HEV in serum.
Highlights
Hepatitis E virus (HEV) causes hepatitis E, which is considered to be a significant public health problem
Expressed HEV ORF2 recombinant protein fused to a His-tag was purified from gels and used to immunize BALB/c mice
Recombinant ORF2 proteins were used to screen positive monoclonal antibodies (mAbs) produced by hybridomas, and the 6×His-tag recombinant protein was used to eliminate the possibility of mAbs reacting with the His-tag protein
Summary
Hepatitis E virus (HEV) causes hepatitis E, which is considered to be a significant public health problem. It spreads in both developing [1] and industrialized countries [2], and is responsible for around 20 million infections and 70 thousand deaths each year according to World Health Organization. HEV is a single-stranded, positive-sense RNA virus classified into the genus. Epitope motifs in HEV ORF2 protein by GZ, http://119.253.58.231/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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