Medium Optimization And Process Development For Stable Cell Line CHO-K1-3E5

Recently,the market of biological drugs has achieved great success.Especially the development of therapeutic antibodies,are becoming a bright spot in pharmaceutical industry.The production of a large amount of antibodies relies on the mammalian cell culture platform in the manufacturing process,which is still a limitation in the development of biopharmaceutical products.The aim of this study was to develop,optimize and validate a high-yield antibody expression platform using"design of experiment" and "process analytical technology" approaches.Previously,we generated the cell pool 160F1,expressing monoclonal antibody 6D11-162,through genetic recombination.In this study,five original cell clones were obtained from the 160F1 pool.After evaluation,one cell line 3E5 was selected with better anti-sheering ability,and was processed to the optimization of the culture media.The number of 17 base media and 7 feed media were screened by detecting and adjusting the amino acids,glucose,lactic acid,ammonia,in the culture supernatants,using the methods of DOE and PAT.It was demonstrated that the best combination is OBM-1 basal medium with the OFM-1 feed medium in this study.The small scale shake flask expression of 6D11-162 in 3E5 cells can reach the final antibody concentration of 1237.13 mg/L,with the cell density of 1.23×107 cells/mL.It was consistant with the preliminary process in 2 L bioreactor,that the final antibody concentration reach to 837.77 mg/L with a cell density of 8× 106 cells/mL.In summary,the cell line 3E5,stablely expressing the mAb 6D11-162,was generated.The optimization and validation of the culture media was carried out based on the 3E5 cells.We has been successfully established a small scale antibody expression platform,and also scaled that up with repeatable and reasonable antibody yield for biopharmaceutical application.