The Chinese Hamster Ovary （CHO K1） cell was used to express a targeted anti-cancer monoclonal antibody by optimizing the platform of the construction of production cell line in this study. The adherent CHO K1 was fi...The Chinese Hamster Ovary （CHO K1） cell was used to express a targeted anti-cancer monoclonal antibody by optimizing the platform of the construction of production cell line in this study. The adherent CHO K1 was first adapted to suspension culture in chemical defined medium. Then the glutamine synthetase （GS） vector was applied to construct a single plasmid to overexpress a monoclonal antibody IgG1. Post transfection, the produc- tion of cell pool was optimized by glutamine-free selection and amplification using various concentrations of methio- nine sulfoximine. The best cell pool ofCHO K1/IgG1 was used to screen the top single clone using the limiting dilution cloning. Finally, a high IgG1 production of 780 mg/L was obtained from a batch culture. This study demonstrated that the construction of high producing cell line, from gene to clone, could be completed within six month and the gene amplification improved protein production greatly.展开更多
文摘The Chinese Hamster Ovary （CHO K1） cell was used to express a targeted anti-cancer monoclonal antibody by optimizing the platform of the construction of production cell line in this study. The adherent CHO K1 was first adapted to suspension culture in chemical defined medium. Then the glutamine synthetase （GS） vector was applied to construct a single plasmid to overexpress a monoclonal antibody IgG1. Post transfection, the produc- tion of cell pool was optimized by glutamine-free selection and amplification using various concentrations of methio- nine sulfoximine. The best cell pool ofCHO K1/IgG1 was used to screen the top single clone using the limiting dilution cloning. Finally, a high IgG1 production of 780 mg/L was obtained from a batch culture. This study demonstrated that the construction of high producing cell line, from gene to clone, could be completed within six month and the gene amplification improved protein production greatly.
