Abstract: To specifically deplete the compound naringin from Si-Ni-San, a traditional Chinese prescription, an immunoaffinity chromatography column was prepared. The naringin bovine serum albumin (BSA) complex (naringin-BSA) was synthesized to make the complete antigen naringin-BSA. Polyclonal antibody was prepared from the serum of rabbit immunized with naringin-BSA. Then, an immunoaffinity column was made by covalently coupling the polyclonal antibody to CNBr-activated Sepharose 4B and used for depleting naringin from Si-Ni-San. The polyclonal antibody obtained from the rabbit serum was found through enzyme linked immunosorbent assay (ELISA) to show the titer of 1∶30000, the purity of 94% and low cross-reaction rate. The coupling rate of the polyclonal antibody to Sepharose 4B was 87%. By using this column, naringin in Si-Ni-San was selectively depleted from the whole extract. This immunoaffinity column of anti-naringin antibody could be used for specifically depleting naringin from Si-Ni-San and other samples.

Key words: CNBr-activated Sepharose 4B , complete antigen, Immunoaffinity column, Naringin, Si-Ni-San, specific depletion