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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Enhanced Sialylation of Albumin-erythropoietin by Biphasic Cultivation in CHO Cells

CHO 세포의 2단계 배양을 통한 Albumin-erythropoietin의 시알산 증대

  • Lim, Jin-Hyuk (Department of Biological Engineering, Inha University) ;
  • Shin, Soo-Ah (Department of Biological Engineering, Inha University) ;
  • Cha, Hyun-Myoung (Department of Biological Engineering, Inha University) ;
  • Kim, Dong-Il (Department of Biological Engineering, Inha University)
  • 임진혁 (인하대학교 공과대학 생물공학과) ;
  • 신수아 (인하대학교 공과대학 생물공학과) ;
  • 차현명 (인하대학교 공과대학 생물공학과) ;
  • 김동일 (인하대학교 공과대학 생물공학과)
  • Received : 2016.08.31
  • Accepted : 2016.12.21
  • Published : 2016.12.31
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Abstract

In glycoprotein, Terminal sialic acid residues of N-linked 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 $CO_2$ 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 $37^{\circ}C$ respectively and the $CO_2$ 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 $CO_2$ and $33^{\circ}C$ on day 5. However, the temperature and concentration of $CO_2$ have little effect on activity of ${\alpha}2,3$-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.

Keywords

References

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