A plant-based transient expression system for the rapid production of highly immunogenic Hepatitis E virus-like particle
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ORIGINAL RESEARCH PAPER
A plant-based transient expression system for the rapid production of highly immunogenic Hepatitis E virus-like particles Eugenia S. Mardanova . Katerina H. Takova . Valentina T. Toneva . Gergana G. Zahmanova . Liudmila M. Tsybalova . Nikolai V. Ravin
Received: 4 June 2020 / Accepted: 25 August 2020 Ó Springer Nature B.V. 2020
Abstract Objective Hepatitis E virus (HEV) infection is a major cause of acute hepatitis worldwide. The aim of the study is the development of plant expression system for the production of virus-like particles formed by HEV capsid and the characterization of their immunogenicity. Results Open reading frame (ORF) 2 encodes the viral capsid protein and possesses candidate for vaccine production. In this study, we used truncated genotype 3 HEV ORF 2 consisting of aa residues 110 to 610. The recombinant protein was expressed in
Nicotiana benthamiana plants using the self-replicating potato virus X-based vector pEff up to 10% of the soluble protein fraction. The yield of HEV 110-610 after purification was 150–200 lg per 1 g of green leaf biomass. The recombinant protein formed nanosized virus-like particles. The immunization of mice with plant-produced HEV 110-610 protein induced high levels of HEV-specific serum antibodies. Conclusions HEV ORF 2 (110-610 aa) can be used as candidate for the development of a plant-produced vaccine against Hepatitis E. Keywords Hepatitis E virus Plant biofactory Potato virus X Vaccine Viral vector
E. S. Mardanova N. V. Ravin (&) Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia e-mail: [email protected] K. H. Takova V. T. Toneva G. G. Zahmanova Department of Plant Physiology and Molecular Biology, University of Plovdiv, 4000 Plovdiv, Bulgaria V. T. Toneva Institute of Molecular Biology and Biotechnologies, 4000 Plovdiv, Bulgaria G. G. Zahmanova Center of Plant Systems Biology and Biotechnology, 4000 Plovdiv, Bulgaria L. M. Tsybalova Research Institute of Influenza, Russian Ministry of Health, St. Petersburg 197376, Russia
Abbreviations HEV Hepatitis E virus ORF Open reading frame PVX Potato virus X
Introduction Hepatitis E virus (HEV) infection is a worldwide disease and the primary cause of acute viral hepatitis in the world with an estimated 20 million cases every year (Larrue et al. 2020). In recent years, increased cases have been reported in developing countries; however sporadic locally acquired cases of HEV
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infections have been also reported in Europe, Australia and Japan (Goel and Aggarwal 2020). Prophylactic vaccination is an effective method to protect against HEV infection and control HEV infection-associated diseases (Li et al. 2020). Hepatitis E virus belongs to the Hepevirida family, genus Orthohepevirus. The genome of HEV is a single single-stranded positive-sense RNA molecule with a length of about 7.3 kb, containing three open reading frames (ORFs) (Tam et al. 199
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