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KMID : 0380020170320020090
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Korean Journal of Biotechnology and Bioengineering
2017 Volume.32 No. 2 p.90 ~ p.95
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Effects of Low-Serum Medium and Various Culture Additives on Production of Recombinant Human Erythropoietin in CHO Cell Cultures
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Lee Kyung-Sun
Cha Hyun-Myoung
Lim Jin-Hyuk
Kim Dong-Il
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Abstract
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Mammalian cell cultures have been used extensively to produce proteins for therapeutic agent because of their ability to perform post-translational modification including glycosylation. To produce recombinant protein, many factors and parameter are considered such as media composition, host cell type, and culture process. In this study, recombinant human erythropoietin (rhEPO) producing cell line was established by using glutamine synthetase system. To reduce serum concentration in media, we compared direct adaptation with step adaptation. Cell growth was faster in step adaptation. In low-level serum media, there were insufficient glucose for cell growth. Thus, we added glucose in low-level serum media from 2 g/L to 4.5 g/L. Titer of rhEPO was higher than other conditions at 4.5 g/L of glucose. Additionally, N-methyl-D-aspartate (NMDA), 13-cis-retinal, and pluronic F-68 (PF-68) were added to enhance productivity in CHO cell cultures. In conclusion, we applied CHO cell producing rhEPO to low-level of serum in media using step-adaptation. Also, we confirmed positive effect of NMDA, 13-cis-retinal, and PF-68.
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KEYWORD
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CHO cell culture, Erythropoietin, Glutamine synthetase, Low-serum media
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