Synthesis of huperzine A-bovine serum albumin conjugate and its immunogenicity in mice

Objective To prepare huperzine A-carrier protein conjugate and determine its immunogenicity for later production of monoclonal antibody against HupA and establishing an enzyme-linked immunosorbent assay ( ELISA) -based detection method for HupA. Methods HupA was conjugated with bovine serum albumin ( BSA) using glutaraldehyde ( GA) method to obtain the HupA-GA-BSA conjugate,and the hapten number in the conjugate was determined by matrix-assisted laser absorption ionization time-of-flight mass spectrometry ( MALDI-TOF-MS) . BALB /c mice were immunized with HupA-GA-BSA to prepare the antiserum against HupA. The serum titer and specificity of the HupA antibodies were detected by indirect ELISA and competitive ELISA,respectively. Results The conjugation ratio of HupA and the carrier protein BSA was 8∶ 1. The antiserum against HupA with a titer of 1∶ 62 500 was obtained after immunizing the mice with the conjugate,and the antiserum reacted specifically to HupA. Conclusion The synthesized HupA-GA-BSA conjugate possesses good immunogenicity in mice,suggesting the feasibility of preparing the monoclonal antibody against HupA using this conjugate.