Purification and characterization of hepatitis B virus surface antigen particles produced in Drosophila Schneider-2 cells.

Abstract:

: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.

journal_name

J Virol Methods

authors

Deml L,Schirmbeck R,Reimann J,Wolf H,Wagner R

doi

10.1016/s0166-0934(99)00022-1

subject

Has Abstract

pub_date

1999-05-01 00:00:00

pages

205-17

issue

2

eissn

0166-0934

issn

1879-0984

pii

S0166093499000221

journal_volume

79

pub_type

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