Effect of sodium butyrate on glycosylation of recombinant erythropoietin

다국어 초록 (Multilingual Abstract)

The effect of Sodium Butyrate(NaBu) on the Nlinked oligosaccharide structure of Erythropoietin(EPO) was investigated. Recombinant human EPO was produced by CHO cells grown in an MEMαmedium with or without 5 mM NaBu, and purified from the culture supernatants using a heparin-sepharose affinity column and immonoaffinity 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) value. A glycan analysis by HPLC showed that the most signigicant characteristic effect of NaBu was a reduction in the proportion of glycans with tri- 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 asialoglycan increased from 12.54% to 23.69% when treated with NaBu.

목차 (Table of Contents)

• MATERIALS AND METHODS
• Cell Line and Culture Conditions
• Purification of Recombinant EPO
• Preparation of N-linked Oligosaccharides from Recombinant EPO
• 2-Aminobenzamide(2-AB) Labeling
• HPLC Analysis of Oligosaccharides
• RESULTS AND DISCUSSION
• Purification of Recombinant EPO
• Release of Oligosaccharides of EPO and 2-Aminobenzamide Labeling
• Comparative Study of Oligosaccharide Structure from EPO Produced in Culture with or without NaBu