Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Abstract

Background

Optimization of chemically defined medium has been a critical way to produce monoclonal antibody. Usually, amount of glutamine was added into the feed medium, but a half of asparagine was added. Our study found that asparagine was important in the antibody production phase. Increasing the ratio of asparagine to glutamine in feed medium for enhancement of antibody production in CHO-DHFR cell culture would be an efficient way.

Results

We optimized the total amount and the ratio of the two vital amino acids in feed medium to increase antibody production. In this work, we have demonstrated that feeding medium of high ratio between asparagine and glutamine (FB-H) can enhance the cell density after reaching the stationary phase. Moreover, FB-H was shown to improve cell maintenance, and increased the antibody production. The metabolic flux analysis proved that ratio of asparagine to glutamine had little influence on glycolysis. Furthermore, the TCA cycle of FB-H was enhanced by 20 % compared to that of low ratio of asparagine to glutamine (FB-L). And the energy metabolism of FB-H was 22.6 % higher than that of FB-L. For the later, lactate can be less produced in FB-H.

Conclusions

We should improve the ratio between asparagine and glutamine in feed medium properly under the premise of no influence on cell growth to achieve high mAb producing goal.

Keywords

Chinese Hamster ovary cells / Feed medium / Ratio between asparagine and glutamine / Cell growth / Monoclonal antibody production / Metabolism

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Funding

the National Natural Science Foundation of China(21206040)

the National High Technology Research and Development Program of China (863 Program)(2012AA02A303)

the Fundamental Research Funds for the Central Universities(WF1214035)