Abstract
Hepatitis E is a globally distributed human disease caused by hepatitis E virus (HEV). In Europe, it spreads through undercooked pork meat or other products and with blood components through transfusions. There are no approved or golden standard serologic systems for HEV diagnostics. Commercially available HEV tests often provide inconsistent results which may differ among the assays. In this study, we describe generation in yeast and characterization of HEV genotype 3 (HEV-3) and rat HEV capsid proteins self-assembled into virus-like particles (VLPs) and the development of HEV-specific monoclonal antibodies (MAbs). Full-length HEV-3 and rat HEV capsid proteins and their truncated variants comprising amino acids (aa) 112-608 were produced in yeast S. cerevisiae. The yeast-expressed rat HEV capsid protein was found to be glycosylated. The full-length HEV-3 capsid protein and both full-length and truncated rat HEV capsid proteins were capable to self-assemble into VLPs. All recombinant proteins contained HEV genotype-specific linear epitopes and cross-reactive conformational epitopes recognized by serum antibodies from HEV-infected reservoir animals. Two panels of MAbs against HEV-3 and rat HEV capsid proteins were generated. Their cross-reactivity pattern was investigated by Western blot, ELISA, and immunofluorescence assay on HEV-3-infected cell cultures. The analysis revealed cross-reactive, genotype-specific, and virus-reactive MAbs. MAb epitopes were localized within S, M, and P domains of HEV-3 and rat HEV capsid proteins. Yeast-generated recombinant VLPs of HEV-3 and rat HEV capsid proteins and HEV-specific MAbs might be employed to develop novel HEV detection systems.
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Acknowledgments
We thank Marius Jasiulionis from the Nature Research Center (Vilnius, Lithuania) who provided wild rat samples.
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This research was funded by the Lithuanian Science Council, Grant No. MIP-039/2015.
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I.K.-K. and M.S. conceived/designed the experiments, analyzed the data, and wrote the paper; M.S., P.L.T, R.P.B., and R.J. performed the research; A.Z., R.J., and R.G.U. contributed materials/analysis tools and revised the manuscript.
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All applicable institutional, national, and international guidelines for the care and use of experimental animals were followed. The mice were used under the license no. LT 59-902, permission no. 184 for breeding of experimental mice, and permission no. 209 for generation of polyclonal and monoclonal antibodies obtained from the State Food and Veterinary Service, Vilnius, Lithuania.
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Simanavicius, M., Tamosiunas, P.L., Petraityte-Burneikiene, R. et al. Generation in yeast and antigenic characterization of hepatitis E virus capsid protein virus-like particles. Appl Microbiol Biotechnol 102, 185–198 (2018). https://doi.org/10.1007/s00253-017-8622-9
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DOI: https://doi.org/10.1007/s00253-017-8622-9