Purification and characterization of hepatitis B virus surface antigen particles produced in Drosophila Schneider-2 cells.
The small surface antigen of hepatitis B virus (HBV) was produced in Drosophila melanogaster Schneider-2 (DS-2) cells transfected stably using an inducible Drosophila metallothionein promoter. Selected clonal DS-2 cell-lines expressed and secreted large quantities of HBsAg particles consisting exclusively of non-glycosylated 25 kDa proteins. HBsAg produced by DS-2 cells had physical and biochemical properties very similar to 22 nm particles derived from the human hepatoma cell-line PLC/PRF/5. DS-2 cell-derived HBsAg particles were purified near homogeneity by a strategy based on protein concentration, precipitation and ultracentrifugation. The resulting HBsAg product was < 98% pure. A single immunisation of BALB/c mice with both DS-2 and yeast-cell derived purified HBsAg particles without adjuvants elicited a substantial humoral antibody and class-I restricted cytotoxic T lymphocyte (CTL) response. Adsorption of HBsAg particles to aluminium hydroxide resulted in increased levels of HBsAg-specific antibodies. However, CTLs were not elicited by HBsAg/Alum combinations. Thus, stably transfected DS-2 cells provide a useful source for the production of HBV subviral particles for diagnostic and research purposes as well as for novel vaccine development.
Deml
L
L
Institute of Medical Microbiology, Klinikum Regensburg, University of Regensburg, Germany.
Schirmbeck
R
R
Reimann
J
J
Wolf
H
H
Wagner
R
R
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004331:Drosophila melanogaster / Q000166:cytology; D005260:Female; D006514:Hepatitis B Surface Antigens / Q000276:immunology* / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification; D014407:Tumor Cells, Cultured; D014771:Virion
D006514:Hepatitis B Surface Antigens; D011993:Recombinant Fusion Proteins