title: Expression and immunogenicity of recombinant polypeptide VP1 of human hepatitis A virus in stably transformed fruitfly (Drosophila melanogaster) Schneider 2 cells.
abstract: We describe the secretory expression and immunogenicity of the recombinant HAV (hepatitis A virus) structural polypeptide VP1 from stably transformed Drosophila melanogaster S2 (Schneider 2) cells. Southern-blot analysis indicated that transformed S2 cells contained multiple copies of the HAV VP1 gene in the genome. Recombinant VP1 was secreted into a culture medium with a molecular mass of 42-49 kDa. A maximum production level of 6.24 mg of recombinant VP1/litre was obtained in a T-flask culture of Drosophila S2 cells 5 days after induction with 0.5 mM CuSO4. The recombinant HAV VP1 protein elicited the production of specific IgA in the small intestine by oral immunization and production of specific IgG in the serum by intraperitoneal immunization. Our findings show that secretory recombinant VP1 from transformed Drosophila S2 cells can be used as an effective experimental immunogen for research in vaccine development.
authors.affiliation: Department of Genetic Engineering, Graduate School of Biotechnology, Kyung Hee University, Suwon 449-701, Korea.
authors.forename: Jong Min
authors.initials: JM
authors.lastname: Lee
authors.forename: Hyun Ho
authors.initials: HH
authors.lastname: Lee
authors.forename: Jeon
authors.initials: J
authors.lastname: Hwang-Bo
authors.forename: Dong Hwa
authors.initials: DH
authors.lastname: Shon
authors.forename: Wonyong
authors.initials: W
authors.lastname: Kim
authors.forename: In Sik
authors.initials: IS
authors.lastname: Chung
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D049109:Cell Proliferation; D004331:Drosophila melanogaster / Q000166:cytology* / Q000235:genetics*; D015870:Gene Expression; D030041:Hepatitis A virus; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D014170:Transformation, Genetic; D015678:Viral Structural Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*
chemical_list: D011994:Recombinant Proteins; C096539:VP1 protein, hepatitis A virus; D015678:Viral Structural Proteins