Fig 3. Reactivity of HBsAg (W1S, W3S and adr4) with a commercial ELISA and an anti-tag Ab.
All the wild-type HBsAgs (W1S, W3S and adr4) were expressed by transient transfection in HEK 293-T cells. The culture supernatants and the cells were collected 72 h after transfection. (A) Reactivity of HBsAg from the culture supernatants and the cell lysates with a commercial ELISA kit. (B) Detection and reactivity of the wild-type HBsAg of PEG-precipitated culture supernatants and cell lysates against anti-Xpress mAb. The expressed HBsAgs were captured on a Ni-coated plate and reacted with an anti-Xpress Ab followed by anti-mouse Ab conjugated with HRP. The reactivity of HBsAgs was presented as the percentage of the OD450-630 of samples compared to W1S. The OD values of HBsAg ELISA and the anti-tag reactivity were normalized by the ratios of densitometric intensity of the Western blotting compared to W1S (Fig 2A). Non-transfected cells served as a negative control (NC). The mean value of three independent experiments is shown as the final reactivity of HBsAg. Error bars are shown as the standard error of means (SEM). wt: wild-type.