Enhancement of recombinant erythropoietin production in CHO cells in an incubator without CO2 addition |
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Authors: | Sung Kwan Yoon Yong-Ho Ahn Kyuboem Han |
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Institution: | (1) Life Science R&D, LG Chemical Investment, Yu Song, Science Town, Daejeon, Korea;(2) Hanson Biotech Co., #201 IACRI, 133 Ojung-dong, Daeduk-gu, Daejeon, Korea |
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Abstract: | The effect of low levels of carbon dioxide (CO2) in the gas phase on the production of recombinant human erythropoietin (EPO)in CHO cells was explored. A T-flask culture
in an incubator without CO2 addition showed a slow cell growth initially followed by the cessation of growth, while other cultures incubated under 0.5–5%
CO2 concentrations grew normally at the same rate during the entire period of cultivation. Interestingly, the production of EPO
in the culture incubated under no CO2 supply was highest among the tested cultures. The cell specific secretion rate of EPO (qEPO) of the culture under no CO2 supply was about 3 times higher than that of the culture under 5% CO2 supply. Western blot analysis and in vivo bioassay of EPO showed no apparent changes in EPO quality between the two cases of different CO2 environments (air vs. 5% CO2), suggesting robust glycosylation of EPO by CHO cells even under very reduced CO2 environment. Various combinations of the two extreme cases, with 5% CO2 supply (suitable for cell growth) and no CO2 addition (better for EPO production), were made in order to maximize the volumetric productivity of EPO secretion (PV) in CHO cells. The PV of the cultures programmed with initial incubation under 5% CO2 followed by no CO2 supply was about 2 times superior to that of the culture incubated only under no CO2 supply. The PV of the culture under no CO2 supply was slightly lower than that of culture grown under 5% CO2. However, the qEPO of the no CO2 supply case was more than 5 times higher than that of the culture under 5% CO2 supply. In conclusion, we have demonstrated that a simple programming of CO2 supply to an incubator can enhance the production of EPO in CHO cells remarkably, without any apparent change of the EPO
quality.
This revised version was published online in August 2006 with corrections to the Cover Date. |
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Keywords: | carbon dioxide CHO cells CO2 EPO erythropoietin mammalian cell culture |
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