Evaluation of the critical process parameters for the cultivation of recombinant Chinese hamster ovary cells in serum-free fed-batch mode.
- Author:
Xingmao LIU
1
;
Hong LIU
;
Lingling YE
;
Shichong LI
;
Benchuan WU
;
Qiwei WANG
;
Zhaolie CHEN
Author Information
1. Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Animals;
CHO Cells;
Cell Culture Techniques;
methods;
Cricetinae;
Cricetulus;
Culture Media, Serum-Free;
Humans;
Recombinant Proteins;
biosynthesis;
genetics;
metabolism;
Urokinase-Type Plasminogen Activator;
biosynthesis;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2011;27(2):240-246
- CountryChina
- Language:Chinese
-
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 x 10(5)-4 x 10(5) cells/mL and the start time of feeding set at 72 h, a maximum viable cells density of 7.8 x 10(6) cells/mL with a peak Pro-UK activity at 8570 IU/mL was achieved during 12 d fed-batch culture. Further, the mu 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.
