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)
分类号：
出版年·卷·期（页码）：2012,32 (4):0-0
DOI:
10.16155/j.0254-1793.2017.01.01
-----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。
-----英文摘要：---------------------------------------------------------------------------------------
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.
-----参考文献：---------------------------------------------------------------------------------------
[1] Schbach JW,Egrie JC,Downing MR,et al.Correction of the anemia of end-stage renal disease with recombinant human erythropoietin.Results of a combined phase Ⅰand Ⅱ clinical trial.N Engl J Med,1987,316(2):73
[2] Abels RI.Use of recombinant human erythropoietin in the treatment of anemia in patients who have cancer.Semin Oncol,1992,19(3 Suppl 8):29
[3] Sasaki H,Bothner B,Dell A,et al.Carbohydrate structure of erythropoietin expressed in Chinese hamster ovary cells by a human erythropoietin cDNA.J Biol Chem,1987,262:12059
[4] Guillermina Forno,Mariela Bollati Fogolin,Marcos Oggero,et al.N-and O-linked carbohydrates and glycosylation site occupancy in recombinant human granulocyte-macrophage colony-stimulating factor secreted by a Chinese hamster ovary cell line.Eur J Biochem FEBS,2004,271:907
[5] Bettina K,Kerstin M,Melanie D.The stability of biopharmaceuticals over time is a primary concern in product development.European Bio Pharmaceutical Review(EBR) SPring,2006,108
[6] Nimtz M,Noll G,Paques E,et al.Conradt,H.S.Carbohydrate structures of human tissue plasminogen activator variant expressed in recombinant Chinese hamster ovary cells.FEBS Lett,1990,271:14
[7] Nimtz M,Martin W,Wray V,et al.Structures of sialylated oligosaccharides of human erythropoietin expressed in recombinant BHK-21 cells.Eur J Biochem,1993,213:39
[8] EP 7.1.2011.97
欢迎阅读《药物分析杂志》！您是该文第 1547位读者！
