Production and Characterization of Self-Assembled Virus-like Particles Comprising Capsid Proteins from Genotypes 3 and 4 Hepatitis E Virus (HEV) and Rabbit HEV Expressed in Escherichia coli .

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101509722 Publication Model: Electronic Cited Medium: Internet ISSN: 1999-4915 (Electronic) Linking ISSN: 19994915 NLM ISO Abbreviation: Viruses Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI

Hepatitis E virus*/genetics ; Hepatitis E virus*/immunology ; Escherichia coli*/genetics ; Escherichia coli*/metabolism ; Genotype* ; Capsid Proteins*/genetics ; Capsid Proteins*/immunology ; Hepatitis E*/immunology ; Hepatitis E*/virology ; Vaccines, Virus-Like Particle*/immunology ; Vaccines, Virus-Like Particle*/genetics; Animals ; Rabbits ; Mice ; Mice, Inbred BALB C ; Viral Hepatitis Vaccines/immunology ; Viral Hepatitis Vaccines/genetics ; Female ; Hepatitis Antibodies/blood ; Hepatitis Antibodies/immunology ; Antibodies, Neutralizing/immunology ; Antibodies, Neutralizing/blood ; Humans ; Antibodies, Viral/immunology ; Antibodies, Viral/blood ; Viral Proteins

The zoonotic transmission of hepatitis E virus (HEV) genotypes 3 (HEV-3) and 4 (HEV-4), and rabbit HEV (HEV-3ra) has been documented. Vaccination against HEV infection depends on the capsid (open reading frame 2, ORF2) protein, which is highly immunogenic and elicits effective virus-neutralizing antibodies. Escherichia coli ( E. coli ) is utilized as an effective system for producing HEV-like particles (VLPs). However, research on the production of ORF2 proteins from these HEV genotypes in E. coli to form VLPs has been modest. In this study, we constructed 21 recombinant plasmids expressing various N-terminally and C-terminally truncated HEV ORF2 proteins for HEV-3, HEV-3ra, and HEV-4 in E. coli . We successfully obtained nine HEV-3, two HEV-3ra, and ten HEV-4 ORF2 proteins, which were primarily localized in inclusion bodies. These proteins were solubilized in 4 M urea, filtered, and subjected to gel filtration. Results revealed that six HEV-3, one HEV-3ra, and two HEV-4 truncated proteins could assemble into VLPs. The purified VLPs displayed molecular weights ranging from 27.1 to 63.4 kDa and demonstrated high purity (74.7-95.3%), as assessed by bioanalyzer, with yields of 13.9-89.6 mg per 100 mL of TB medium. Immunoelectron microscopy confirmed the origin of these VLPs from HEV ORF2. Antigenicity testing indicated that these VLPs possess characteristic HEV antigenicity. Evaluation of immunogenicity in Balb/cAJcl mice revealed robust anti-HEV IgG responses, highlighting the potential of these VLPs as immunogens. These findings suggest that the generated HEV VLPs of different genotypes could serve as valuable tools for HEV research and vaccine development.

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JP22fk0210075 and JP24fk0210132 Japan Agency for Medical Research and Development

Keywords: Escherichia coli; assembly in vitro; capsid protein; genotype; hepatitis E virus; virus-like particle

0 (Capsid Proteins)
0 (Vaccines, Virus-Like Particle)
0 (ORF2 protein, Hepatitis E virus)
0 (Viral Hepatitis Vaccines)
0 (Hepatitis Antibodies)
0 (Antibodies, Neutralizing)
0 (Antibodies, Viral)
0 (Viral Proteins)

Date Created: 20240928 Date Completed: 20240928 Latest Revision: 20240930

20240930

PMC11437457

10.3390/v16091400

39339876