Evaluation of the critical process parameters for the cultivation of recombinant Chinese hamster ovary cells in serum-free fed-batch mode

Abstract:

Taking a suspension adapted recombinant CHO cell line, 11G-S expressing human Pro-urokinase (Pro-UK) as the object of study, the impacts of different feeding nutrients, the start time of feeding and cell inoculation density on the growth and Pro-UK production of 11G-S cells in serum-free fed-batch culture were evaluated in 100 mL shacking flasks. The results indicated that amino acids, serum-free supplements and inorganic salts played important role in cell growth, cell viability and protein expression. And the effects of cells fed-batch culture was much better with the initial cell inoculation density at 3×10⁵~4×10⁵ cells/mL and the start time of feeding set at 72 h, a maximum viable cells density of 7.8×10⁶ cells/mL with a peak Pro-UK activity at 8 570 IU/mL was achieved during 12 d fed-batch culture. Further, the μ of the 11G-S cells at the middle phase of the fed-batch culture, and both the q_glu and q_gln of the 11G-S cells at the middle and later phases of the fed-batch culture was higher than that of the 11G-S cells at the same phase of the batch culture, respectively.