Microscale nonreductive release of O-linked glycans for subsequent analysis through MALDI mass spectrometry and capillary electrophoresis
Microscale nonreductive release of O-linked glycans for subsequent analysis through MALDI mass spectrometry and capillary electrophoresis
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DOI：
10.1021/ac015534c
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发表时间：
2001-12-15
影响因子：
7.4
通讯作者：
Novotny, MV
中科院分区：
文献类型：
--
作者：
Huang, YP;Mechref, Y;Novotny, MV
已经设计了一种新的基于β-消除的程序，用于从糖蛋白中微量释放O-连接的寡糖。与传统的卡尔森降解，这导致糖醇的形成，在这里报告的程序，使还原端完整。在氨介质中将释放的寡糖转化为葡糖胺，然后通过加入硼酸产生还原性寡糖。定量生成的具有还原端的寡糖随后可以用荧光试剂衍生化用于毛细管电泳，或者通过MALDI质谱法进行分析。这些化学步骤的微观版本使我们能够在非常低的水平上研究结构上的O-连接寡糖。
A new beta -elimination-based procedure has been devised for a microscale release of O-linked oligosaccharides from glycoproteins. Unlike the conventional Carlson degradation, which leads to formation of alditols, the procedure reported here renders the reducing end intact. Conversion of the liberated oligosaccharides to glycosylamines, in ammonia medium is followed by the production of the reducing oligosaccharides through the addition of boric acid. The quantitatively generated oligosaccharides with the reducing end can subsequently be derivatized with a fluorophoric reagent for capillary electrophoresis or, alternatively, analyzed through MALDI mass spectrometry. The microscale version of these chemical steps permits us to investigate structurally O-linked oligosaccharides at very low levels.