|
KMID : 0545120130230050699
|
|
Journal of Microbiology and Biotechnology
2013 Volume.23 No. 5 p.699 ~ p.706
|
|
|
Effect of Mild-Thiol Reducing Agents and α2,3-Sialyltransferase Expression on Secretion and Sialylation of Recombinant EPO in CHO Cells
|
|
Chang Kern-Hee
Jeong Yeon-Tae
Kwak Chan-Yeong
Choi One
Kim Jung-Hoe
|
|
|
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 thiolreducing 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 α2,3-sialyltransferase (α2,3-ST) was introduced into EPO-producing CHO cells in order to compensate for the reduced sialylation during supplementation with NAC. When α2,3-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.
|
|
|
KEYWORD
|
|
|
CHO cells, Erythropoietin, Sialylation, Sialyltransferase, Thiol reducing agents
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|
|