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KMID : 0380020190340020073
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Korean Journal of Biotechnology and Bioengineering
2019 Volume.34 No. 2 p.73 ~ p.79
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Enhanced Sialylation of Albumin-erythropoietin by Controlling Hexose Combination Ratio in Chinese Hamster Ovary Cell Cultures
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Kim Na-Hee
Cha Hyun-Myoung
Lim Jin-Hyuk
Lee Ji-Hun
Han Hye-Jin
Kim Dong-Il
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Abstract
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Sialylation is very important for in vivo half-life of therapeutic proteins because sialic acids can reduce the rates of clearance in hepatocyte. Many studies were developed for supplementing various carbon sources which directly influcenced cellular metabolism and glycosylation. So it is necessary to investigate interconversion of carbons sources in combination because once they are absorbed into cells and synthesized into nucleotide sugars, they can be converted to different kinds of sugars and nucleotide sugars. Albumin-erythropoietin (Alb-EPO) has three glycan sites with tetra-antennary structure, so it can be added up to 14 sialic acids. Therefore, we demonstrated the effects of combination ratios of hexose (glucose:galactose:mannose) among the generally used carbon sources essential for cell growth on sialylation of a fusion protein, Alb-EPO. As a result, there was no negative effect on cell growth except the condition of the highest concentration. The titer of Alb-EPO was increased depending on increasing concentration of hexoses in order of mannose, glucose, and galactose and reached up to 1.7-fold in comparison to the control. On the other hand, there was more than 1.5-fold increase in sialylation followed by changed nucleotide sugar pool at the conditions supplemented with glucose and galactose at high concentration. In conclusion, our strategy of controlling the concentrations of available carbon sources offers an effective approach for the improved production of highly sialylated Alb-EPO without deleterious effect on growth.
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KEYWORD
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albumin-erythropoietin, CHO cell, hexose, sialylation
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