Generation of Anti-HBc Monoclonal Antibodies

Immunization of BALB/c mice and generation of hybridomas producing anti-HBc mAbs were carried out as previously described [28 (link)]. Briefly, purified His-precore/core was injected into BALB/c mice using keyhole limpet hemocyanin as a carrier protein. Four weeks later, lymphocytes were isolated and fused to myeloma cell. Isotype determination was then performed using the IsoStrip Mouse Monoclonal Antibody Isotyping Kit according to the manufacturer’s protocol (Roche Diagnostics, Basel, Switzerland). Purification of mAbs from hybridoma supernatant was performed using a Spin column based Antibody Purification Kit (Protein G) (Cosmo Bio Co., Ltd., Tokyo, Japan). The protein concentration was calculated using a NanoDrop 2000/2000c spectrophotometer (Thermo Fisher Scientific).