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)

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.