Simultaneous microanalysis of N-linked oligosaccharides in a glycoprotein using microbore graphitized carbon column liquid chromatography-mass spectrometry
Simultaneous microanalysis of N-linked oligosaccharides in a glycoprotein using microbore graphitized carbon column liquid chromatography-mass spectrometry
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DOI：
10.1016/s0021-9673(02)00951-2
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发表时间：
2002-08-30
影响因子：
4.1
通讯作者：
Hayakawa, T
中科院分区：
文献类型：
--
作者：
Itoh, S;Kawasaki, N;Hayakawa, T
我们以前报道过石墨化碳柱液质联用(GCC-LC-MS)对于糖蛋白中碳水化合物的结构分析是非常有用的。本研究采用GCC-LC-MS联用技术同时测定低聚糖的含量。以胎球蛋白、核糖核酸酶B和重组人促红细胞生成素为模型寡糖制备了多种寡糖糖醇。利用微孔GCC-LC-MS可以快速、灵敏地同时分析高甘露糖型、岩藻糖络合型、唾液酸化复合型和唾液酸岩藻糖络合型寡糖糖醇。此外，我们还证明了该方法适用于具有不同寡糖的糖蛋白中碳水化合物的异质性分析。微孔GCC-LC-MS能够在一次分析中表征从人黑色素瘤细胞产生的组织型纤溶酶原激活剂中的高甘露糖型、杂交型和复合型寡糖。(C)2002 Elsevier Science B.V.保留所有权利。
We previously reported that graphitized carbon column liquid chromatography-mass spectrometry (GCC-LC-MS) is very useful for the structural analysis of carbohydrates in a glycoprotein. In this study, GCC-LC-MS was adapted for the simultaneous microanalysis of oligosaccharides. A variety of oligosaccharide alditols prepared from fetuin, ribonuclease B, and recombinant human erythropoietin were used as model oligosaccharides. The use of microbore GCC-LC-MS was found to be successful for rapid, sensitive, and simultaneous analysis of high-mannose-type, desialylated fucosyl complex-type, sialylated complex-type, and sialylated fucosyl complex-type oligosaccharide alditols. Furthermore, we demonstrate that this method is applicable to the analysis of carbohydrate heterogeneity in a glycoprotein that possesses diverse oligosaccharides. Microbore GCC-LC-MS was able to characterize high-mannose-type, hybrid-type, and complex-type oligosaccharides in tissue plasminogen activator produced from human melanoma cells in a single analysis. (C) 2002 Elsevier Science B.V. All rights reserved.