Abstract: In this study, hybridoma cell was adapted to the mixed medium containing less fetal bovine serum through suspension domestication, and the antibody concentration increased largely. The components of mixed medium were RPM1640∶ SFM4CHO∶ OptiCHO(4.5∶ 4.5∶ 1) with 2% FBS and the fed-batch fermentation process was carried out, in which the initial inoculation concentration was 3×105 cells/mL, and 3% (initial volume) chemical-defined feed medium was added every other day during cultivation. The antibody concentration increased to 168 mg/L from 84 mg/L. The hybridoma cells were further cultured in a 5 L stirred-bioreactor, and the products were purified by protein A affinity chromatography and the purified antibody concentration was 240 mg/L.

Key words: hybridoma cell culture, monoclonal antibody, suspension domestication, affinity chromatography