A plant-based transient expression system for the rapid production of highly immunogenic Hepatitis E virus-like particles

Biotechnol Lett. 2020 Nov;42(11):2441-2446. doi: 10.1007/s10529-020-02995-x. Epub 2020 Sep 1.


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 µg 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.

MeSH terms

  • Animals
  • Female
  • Hepatitis Antibodies / blood
  • Hepatitis E / immunology
  • Hepatitis E / prevention & control*
  • Hepatitis E virus / immunology*
  • Hepatitis E virus / metabolism
  • Immunization
  • Influenza Vaccines / administration & dosage*
  • Influenza Vaccines / immunology
  • Influenza Vaccines / metabolism
  • Injections, Intramuscular
  • Mice
  • Mutation*
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Tobacco / genetics
  • Tobacco / growth & development*
  • Viral Proteins / genetics*
  • Viral Proteins / immunology


  • Hepatitis Antibodies
  • Influenza Vaccines
  • ORF2 protein, Hepatitis E virus
  • Recombinant Proteins
  • Viral Proteins