Travelling wave ion mobility and negative ion fragmentation for the structural determination of N-linked glycans
Travelling wave ion mobility and negative ion fragmentation for the structural determination of N-linked glycans
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DOI：
10.1002/elps.201200669
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发表时间：
2013-08-01
期刊：
影响因子：
2.9
通讯作者：
Scrivens, James H.
中科院分区：
文献类型：
--
作者：
Harvey, David J.;Scarff, Charlotte A.;Scrivens, James H.
研究了行波离子迁移率分离N-聚糖与其他化合物的能力以及异构体的拆分。带电荷的聚糖，例如从牛胎球蛋白中释放并通过电喷雾电离的唾液酸化复合N-聚糖，可以与残留糖肽分离，从而检测到次要的、更高度唾液酸化的化合物，其中它们的离子被来自不同电荷状态的其他化合物的离子遮蔽。还发现该技术对于从受污染的样品中提取N-聚糖谱是极好的。通过负离子CID片段化进行聚糖的结构鉴定，该方法提供了丰富的结构诊断离子。然而，碎片离子也可能出现在聚糖谱中，在那里它们可能被误认为是聚糖分子离子。碎片和分子离子经常被证明有不同的漂移时间曲线，使他们能够区分。发现一些异构体分离，但仅限于最小的化合物。通过绘制碎片的漂移时间曲线来实现与构象异构体的区分;这些曲线与构象异构体存在的前体离子的曲线相匹配。该技术被应用于调查的N-聚糖释放的真菌Piptoporus betulinus，该技术被用来分离不同的碳水化合物类型存在于生物提取物。
Travelling wave ion mobility was investigated for its ability to separate N-glycans from other compounds and for resolution of isomers. Charged glycans, exemplified by sialylated complex N-glycans released from bovine fetuin and ionised by electrospray, could be separated from residual glycopeptides allowing the minor, more highly sialylated compounds to be detected where their ions were obscured by ions from other compounds in different charge states. This technique was also found to be excellent for extracting the N-glycan profiles from contaminated samples. Structural identification of the glycans was performed by negative ion CID fragmentation, a method that provides a wealth of structurally diagnostic ions. However, fragment ions can also appear in the glycan profiles where they can be mistaken for glycan molecular ions. Fragments and molecular ions were frequently shown to have different drift time profiles, allowing them to be differentiated. Some separation of isomers was found but only for the smallest compounds. Differentiation from conformers was achieved by plotting drift time profiles of the fragments; these profiles matched those of the precursor ions where conformers were present. The techniques were applied to investigations of N-glycans released from the fungus Piptoporus betulinus where the technique was used to separate different carbohydrate types present in biological extracts.