Comparison between HBsAg purified by anti-HBs-coated microparticles and sucrose-cushion purified HBsAg from differently cultured hepatoma-cell-lines.

Constitutively HBsAg expressing HepG3-cells were cultured in serum-free and serum-supplemented medium. After 5 days of cultivation HBsAg was concentrated, subjected to 15% SDS-PAGE and analyzed by silver staining (A) or immunoblotting with HBsAg-specific antibodies (B). M: Molecular weight standard; Lane 1: Commercially available human plasma reactive for HBsAg as positive control (Abbott IMx HBsAg Control No. 2228–10); Lane 2: HBsAg purified from recombinant vaccinia virus-infected HuH-7 cells by anti-HBs-coated microparticles (Abbott IMx HBsAg (V2) Reagent Pack No. 2228–21). Lane 3 and 4: Two fractions of HBsAg expressed by HepG3-cells under serum-free conditions. Lane 5 and 6: Two fractions of HBsAg expressed by HepG3-cells in serum-supplemented media. Protein samples were loaded on one gel. Two non-relevant lanes were cut out of the silver-stain (A) and Western-blot (B) images, as indicated. The positions of anti-HBsAg antibody reactive proteins are depicted on the right.