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A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen.

ABSTRACT: To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)-specific monoclonal antibody (mAb), the Escherichia coli-expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The mAb G117-AA4 of IgG1 isotype was obtained showing a strong reactivity with the homologous E. coli, but also yeast-expressed capsid protein of HEV-3. The mAb strongly cross-reacted with ratHEV capsid protein derivatives produced in both expression systems and weaker with an E. coli-expressed batHEV capsid protein fragment. In addition, the mAb reacted with capsid protein derivatives of genotypes HEV-2 and HEV-4 and common vole hepatitis E virus (cvHEV), produced by the cell-free synthesis in Chinese hamster ovary (CHO) and Spodoptera frugiperda (Sf21) cell lysates. Western blot and line blot reactivity of the mAb with capsid protein derivatives of HEV-1 to HEV-4, cvHEV, ratHEV and batHEV suggested a linear epitope. Use of truncated derivatives of ratHEV capsid protein in ELISA, Western blot, and a Pepscan analysis allowed to map the epitope within a partially surface-exposed region with the amino acid sequence LYTSV. The mAb was also shown to bind to human patient-derived HEV-3 from infected cell culture and to hare HEV-3 and camel HEV-7 capsid proteins from transfected cells by immunofluorescence assay. The novel mAb may serve as a useful tool for further investigations on the pathogenesis of HEV infections and might be used for diagnostic purposes. KEY POINTS: • The antibody showed cross-reactivity with capsid proteins of different hepeviruses. • The linear epitope of the antibody was mapped in a partially surface-exposed region. • The antibody detected native HEV-3 antigen in infected mammalian cells.

SUBMITTER: Kubickova B

PROVIDER: S-EPMC8236046 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen.

Kubickova Barbara B Schenk Jörg A JA Ramm Franziska F Markuškienė Kornelija K Reetz Jochen J Dremsek Paul P Tamosiunas Paulius Lukas PL Cepulyte Laima L Trinh Hoai Anh HA Scholz Johannes J Memczak Henry H Hovestädt Marc M Ryll René R Petraityte-Burneikiene Rasa R Corman Victor M VM Andersson Anika A Becher Dietmar D Groschup Martin H MH Kubick Stefan S Sellrie Frank F Johne Reimar R Ulrich Rainer G RG

Applied microbiology and biotechnology 20210615 12

To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)-specific monoclonal antibody (mAb), the Escherichia coli-expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The mAb G117-AA4 of IgG1 isotype was obtained showing a strong reactivity with the homologous E. coli, but also yeast-expressed capsid protein of HEV-3. The mAb strongly cross-reacted with ratHEV capsid protein de ...[more]

PMID: 34129082

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Dataset Information

A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen.

Publications

A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen.

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