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KMID : 0380020160310040270
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Korean Journal of Biotechnology and Bioengineering
2016 Volume.31 No. 4 p.270 ~ p.276
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Enhanced Sialylation of Albumin-erythropoietin by Biphasic Cultivation in CHO Cells
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Lim Jin-Hyuk
Shin Soo-Ah
Cha Hyun-Myoung
Kim Dong-Il
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Abstract
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In glycoprotein, Terminal sialic acid residues of Nlinked glycan are imperative things because they prevent the recognition from asialoglycoprotein-receptor that affect the half-life of glycoproteins. So establishment of culture process for enhancing sialic acid is important to maximize sialic acid contents of glycoprotein. In this study, we investigated effects of biphasic culture of Chinese hamster ovary (CHO) cells producing albumin-erythropoietin to increase sialylation. Biphasic cultures were performed with shift of CO2 concentrations and temperatures at day 5 when viable cell density was decreased and sialidase was started to be released by cell lysis. The examined temperature set points were 33, 35 and 37oC respectively and the CO2 concentration was 1, 5, 10 and 15%. We confirmed that sialidase activity was the lowest in biphasic culture that was shifted from normal culture condition to 1% of CO2 and 33oC on day 5. However, the temperature and concentration of CO2 have little effect on activity of α2,3-sialyltransferase. Also, sialic acid contents were enhanced 1.13-fold higher than that in control culture. In conclusion, Biphasic cultivation in CHO cells led to inhibition of sialidase activity and increases of sialylated glycan.
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KEYWORD
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Albumin-erythropoietin, Biphasic culture, Chinese hamster ovary cells, Glycosylation, Sialidase
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