Figure 3.

Synthesis and secretion of surface proteins by wild-type and mutant constructs on Huh7 cells. A: The representative constructs of wild-type (WT), Δ1, and Δ2 deletion. B: Western blot analysis of the intracellular large, middle, and small surface proteins. Protein lysates were harvested in 48 hours after transfection and resolved on SDS-PAGE followed by antibody hybridization. The top panels labeled by gp42/p39 represent glycosylated and unglycosylated large surface proteins detected by anti-pre-S1 antibody, MA18/7. The relative amount of large surface protein for pre-S mutants is ∼40% higher than the wild type. The middle panels labeled by gp36/gp33 represent glycosylated, middle surface proteins detected by anti-pre-S2 antibody. The bottom panels represent small surface proteins (gp27/p24). The relative amounts of each study were shown below. C: Relative extracellular secretion of HBsAg by the cloned wild-type and mutant constructs. The culture medium was collected 48 hours after transfection and the supernatants containing HBsAg were assayed by enzyme-linked immunosorbent assay test. The relative amounts of HBsAg secretion between different expression clones are shown.