Effect of Mild-Thiol Reducing Agents and alpha 2,3-Sialyltransferase Expression on Secretion and Sialylation of Recombinant EPO in CHO Cells

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We have previously reported that N-acetylcysteine (NAC) not only delayed apoptosis but also enhanced the production of recombinant erythropoietin (EPO) in Chinese hamster ovary (CHO) cell culture. To investigate the production enhancement mechanism, the effects of similar thiol-reducing agents were studied. Intriguingly, all mild reducing agents examined including mercaptoethanesulfonic acid (MESNA), thiolactic acid (TLA), and thioglycolate (TG) were shown to block apoptosis and increase EPO production. A pulse-chase study of EPO secretion revealed that all four thiol-reducing agents increased the EPO secretion rate; among them TLA showed the highest rate. In terms of product quality, the sialic acid content of the glycoprotein is one of the most important factors. It was reported that a number of glycoproteins produced by CHO cells often have incomplete sialylation, particularly under high-producing conditions. Human alpha 2,3-sialyltransferase (alpha 2,3-ST) was introduced into EPO-producing CHO cells in order to compensate for the reduced sialylation during supplementation with NAC. When alpha 2,3-ST was expressed in the presence of NAC, reduced sialylation was restored and an even more sialylated EPO was produced. Thus, our study is significant in that it offers increased EPO production while still allowing the prevention of decreased sialylation of EPO.
Publisher
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
Issue Date
2013-05
Language
English
Article Type
Article
Keywords

HAMSTER OVARY CELLS; INTERFERON-GAMMA; SODIUM-BUTYRATE; ENHANCED SIALYLATION; SUGAR CHAINS; ERYTHROPOIETIN; GLYCOPROTEINS; CULTURE; DEATH; ALPHA-2,6-SIALYLTRANSFERASE

Citation

JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.23, no.5, pp.699 - 706

ISSN
1017-7825
DOI
10.4014/jmb.1303.03046
URI
http://hdl.handle.net/10203/174037
Appears in Collection
BS-Journal Papers(저널논문)
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