文章基本信息

标题：Structural Characterization of Recombinant Human Erythropoietins by Fluorophore-Assisted Carbonydrate Electrophoresis
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作者：Kazushige MORIMOTO ; Nozomi MAEDA ; Abdel-Alim Foad ABDEL-ALIM 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：1999
卷号：22
期号：1
页码：5-10
DOI：10.1248/bpb.22.5
出版社：The Pharmaceutical Society of Japan
摘要：Routine analysis of the carbohydrate moieties of a glycoprotein is critical for ensuring the consistene quality of biopharmaceutical products. Fluorophore-assisted carbohydrate electrophoresis (FACE) is a recently introduced method for the separation, by polyacrylamide gel electrophoresis, of oligosaccharides labeled with 8-amino-naphthalene-1, 3, 6-trisulphonic acid (ANTS).In this study, we have evaluated the applicability of the FACE method to analysis of the carbohydrate moieties of three different recombinant human erythropoietins (rHuEPOs), two Chinese hamster ovary (CHO) cell-derived rHuEPOs, and one baby hamster kidney (BHK) cell-derived rHuEPO. The N-linked oligosaccharides released from the rHuEPOs were labeled with ANTS. Enzymic sequence analysis of the N-linked oligosaccharides was performed by the FACE method. The results showed that the FACE method was useful for the analysis of sialo-, asialo-, and agalacto-oligosaccharides in the same gel. Its usefulness for rapidly and reliably revealing the oligosaccharide profiles of given glycoproteins, and its reproducibility were also confirmed in this study. In conclusion, the method can be used for evaluating the quality consistency of recombinant glycoprotein products in terms of the carbohydrate moiety.
关键词：recombinant human erythropoietin;FACE;N-linked oligosaccharide;sequencing;exoglycosidase digestion