title: Expression and immunogenic analysis of recombinant polypeptides derived from capsid protein VP1 for developing subunit vaccine material against hepatitis A virus.
abstract: Three recombinant polypeptides, VP1-His, VP1-3N-His, and 3D2-His, were produced by Escherichia coli expression system. Recombinant VP1-His, VP1-3N-His, and 3D2-His were expressed as bands with molecular weights of 32, 38, and 30 kDa, respectively. These were purified by affinity chromatography using Ni-NTA Fast-flow resin and/or ion-exchange chromatography using DEAE-Sepharose Fast-flow resin. Intraperitoneal immunizations of recombinant polypeptides successfully elicited the productions of VP1-His, VP1-3N-His, and 3D2-His specific IgG antibodies (IgG subclass distribution of IgG1>IgG2a>IgG2b>IgG3) in sera and induced the secretions of cytokines IFN-γ and IL-6 in spleen cells. Sera from recombinant VP1-His-, VP1-3N-His-, and 3D2-His-immunized mice neutralized the propagation of HAV. The highest neutralizing activity was shown in sera from recombinant VP1-3N-His-immunized mice. These results suggest that recombinant VP1-3N-His can be a useful source for developing hepatitis A virus (HAV) subunit vaccine candidates.
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Kyoung Ok
authors.initials: KO
authors.lastname: Jang
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Jong-Hwa
authors.initials: JH
authors.lastname: Park
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Hyun Ho
authors.initials: HH
authors.lastname: Lee
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea; RNA Inc., Yongin 446-701, Republic of Korea.
authors.forename: Dae Kyun
authors.initials: DK
authors.lastname: Chung
authors.affiliation: Department of Microbiology and Research Center for Medical Sciences, Chung-Ang University College of Medicine, Seoul 156-756, Republic of Korea.
authors.forename: Wonyong
authors.initials: W
authors.lastname: Kim
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea. Electronic address: ischung@khu.ac.kr.
authors.forename: In Sik
authors.initials: IS
authors.lastname: Chung
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006506:Hepatitis A / Q000097:blood / Q000276:immunology / Q000517:prevention & control*; D030041:Hepatitis A virus / Q000276:immunology*; D007114:Immunization; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D013154:Spleen / Q000166:cytology / Q000276:immunology; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D007074:Immunoglobulin G; D015850:Interleukin-6; D011994:Recombinant Proteins; D015678:Viral Structural Proteins; D014765:Viral Vaccines; D007371:Interferon-gamma
keywords: Antigenicity; Capsid protein VP1; Hepatitis A; Hepatitis A virus; Neutralizing activity; Recombinant polypeptides