“…Even though clinical specimens constitute the best material for evaluating the ability of immunoassays in detecting HBsAg variants, the difficulty in getting such samples with natural HBsAg substitutions usually leads to the use of recombinant S protein variants as substitutes to investigate the performance of the immunoassays in detecting altered HBsAg epitopes. Since the in vitro production of recombinant HBsAg variants can be affected by the substitutions, 14 it is important to carefully normalize the level protein variants. This was achieved in our study by calibrating HBsAg variant proteins with a tag-HA.…”