重组人促红素糖指纹图谱分析技术研究  

Study on methods of glycans for erythropoietin(EPO)

作　　者：周勇[1] 邵宝珠[2] YUEN Chun-ting 王丽[1] 王军志[1] 

机构地区：[1]中国食品药品检定研究院,北京100050 [2]沈阳三生制药有限责任公司,沈阳110141 [3]National Institute for Biological Standards and Control ( NIBSC), Potters Bar, Herts. EN6 3 QG, UK

出　　处：《药物分析杂志》2012年第4期587-591,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：国家“重大新药创制”科技重大专项项目(2009ZX09307-001)

摘　　要：目的:建立重组人促红素N-糖链特征性糖指纹图谱检测分析技术。方法:分别用外切糖苷酶(PNGase F)和神经氨酸酶酶切重组人促红素,超滤离心获得糖的各种组分,使用高pH离子色谱分析N-糖链的结构和组成并确定N-糖链指纹图谱。结果:建立的方法稳定可靠,重复性好,可用于重组人促红素糖结构和组成的研究和质量控制。结论:本研究建立了简便、有效的重组人促红素N-糖链特征性糖指纹图谱检测分析方法。Objective：To establish analytical methods for N-glycans finger printing of human recombinant erythropoietin（rHuEPO）.Methods：The N-glycosidically-linked oligosaccharide chains and the terminal sialic acid residues of rHuEPO are released from the glycoprotein by N-glycosidase F and neuraminidase digestion separately.After removal of the protein fraction,the resultiant oligosaccharides are chromatographically separated using high pH anion-exchange chromatography with pulsed amperometric detection（HPAEC-PAD）.Results：The methods are well suited for the study and quality control of N-glycans structure and composition of rHuEPO,and the results of all tests were reliable and reproducible.Conclusion：The analytical methods of characteristic N-glycans finger printing have been established for the analysis and quality control of rHuEPO N-glycans.

关 键 词：糖蛋白激素 重组人促红素(rHuEPO) 糖指纹图谱 特征图谱 酶切图谱 糖谱分析技术 

分 类 号：R917[医药卫生—药物分析学]