Monolisa hbeag ab plus kit

HepG2^hNTCP cells were seeded in 48‐well plates 1 day prior to HBV infection. Sera from mice immunized with adjuvant or the plant‐produced HBV‐S/preS1^16‐42 antigen were collected at day 49 post‐immunization and diluted 1:50 in complete DMEM media supplemented with 4% polyethylene glycol (PEG, Sigma Aldrich). Pre‐immune sera from individual mice were pooled per mice group and diluted in the same manner. WT HBV and the vaccine escape HBV‐S^G145R (100 genome equivalents/cell) were incubated with diluted sera for 1 h at 37 °C, before infection. Control conditions included cell treatment with 1 μm Myrcludex B (Pepscan), a well‐established inhibitor of HBV entry (Urban et al., 2014 (link)), for 3 h prior to virus inoculation and cells infected with untreated HBV. At 16 h post‐infection, cells were extensively washed with PBS and incubated with complete DMEM media, supplemented with 2.5% DMSO (AppliChem). The medium collected between days 7–11 post‐infection was retained for quantification of the HBeAg by using the Monolisa HBe Ag‐Ab PLUS kit (BioRad). Inhibition of HBV infection by the immune sera was represented as percentage from infectivity values in the presence of the pre‐immune sera, at the same dilution. Data were statistically analysed by using the Mann–Whitney U test in GraphPad Prism version 6.