Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Effect of Sodium Bytyrate on Glycosylation of Recombinant Erythropoietin

  • Chung, Bo-Sup (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) ;
  • Chang, Kern-hee (Department of Biological Sciences, Korea Advanced Institute of Science and Technology) ;
  • Kim, Jong-Soo (Department of Biological Sciences, Korea Advanced Institute of Science and Technology)
  • Published : 2001.12.01
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Abstract

The effect of Sodium Butyrate (NaBu) on the N-linked oligosaccharide structure of Erythropoietin (EPO) was investigated. Recombinant human EPO was produced by CHO cells grown in an $MEM{\alpha}$ medium with or without 5 mM NaBu, and purified from the culture supernatants using a heparin-sepharose affinity column and immunoaffinity column. The N-linked oligosaccharides were released enzymatically and isolated by paper chromatography. The isolated oligosaccharides were then labeled with a fluorescent dye, 2-aminobenzamide, and analyzed with MonoQ anion exchange chromatography and GlycosepN amide chromatography for the assignment of a GU (glucose unit) vague. A glycan analysis by HPLC showed that the most significant characteristic effect of NaBu was a reduction in the proportion of glycans with Sri-and tetrasialylated oligodaccharides from $21.30\%$ (tri-) and $14.86\%$ (tetra-) in the control cultures (without NaBu) to $8.72\%$ (tri-) and $1.25\%$ (tetra-) in the NaBu-treated cultures, respectively. It was also found that the proportion of asialo-glycan increased from $12.54\%\;to\;23.6\%$ when treated with NaBu.

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