Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Effect of Mild-Thiol Reducing Agents and ${\alpha}2,3$-Sialyltransferase Expression on Secretion and Sialylation of Recombinant EPO in CHO Cells

  • Chang, Kern Hee (Department of Biological Sciences, Korea Advanced Institute of Science and Technology) ;
  • Jeong, Yeon Tae (Department of Biological Sciences, Korea Advanced Institute of Science and Technology) ;
  • Kwak, Chan Yeong (Department of Biological Sciences, Korea Advanced Institute of Science and Technology) ;
  • Choi, One (Department of Biological Sciences, Korea Advanced Institute of Science and Technology) ;
  • Kim, Jung Hoe (Department of Biological Sciences, Korea Advanced Institute of Science and Technology)
  • Received : 2013.03.13
  • Accepted : 2013.03.14
  • Published : 2013.05.28
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Abstract

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.

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References

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