Expression and immunogenicity of recombinant polypeptide VP1 of human hepatitis A virus in stably transformed fruitfly (Drosophila melanogaster) Schneider 2 cells.
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.
Lee
Jong Min
JM
Department of Genetic Engineering, Graduate School of Biotechnology, Kyung Hee University, Suwon 449-701, Korea.
Lee
Hyun Ho
HH
Hwang-Bo
Jeon
J
Shon
Dong Hwa
DH
Kim
Wonyong
W
Chung
In Sik
IS
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*
D011994:Recombinant Proteins; C096539:VP1 protein, hepatitis A virus; D015678:Viral Structural Proteins