Sample preparation for mass spectrometric analysis of human serum N-glycans using hydrophilic interaction chromatography-based solid phase extraction
Sample preparation for mass spectrometric analysis of human serum N-glycans using hydrophilic interaction chromatography-based solid phase extraction
复制标题
使用基于亲水相互作用色谱的固相萃取进行人血清 N-聚糖质谱分析的样品制备
DOI：
10.1039/c4an00660g
复制
发表时间：
2014-09-21
期刊：
影响因子：
4.2
通讯作者：
Liang, Xinmiao
中科院分区：
文献类型：
--
作者：
Cao, Liwei;Zhang, Ye;Liang, Xinmiao
已显示来源于人血清糖蛋白的N-连接聚糖的表达水平在许多疾病的进展期间发生变化。通常，从人血清蛋白中释放的N-聚糖与内源性血清肽、盐和其他污染物共存。有效去除这些污染物对于获得人血清蛋白的聚糖谱是必不可少的。在这里，我们开发了一种样品制备方法，用于质谱（MS）分析的N-连接聚糖来自人血清糖蛋白的基础上的两性离子亲水性材料命名为点击TE-Cys。Click TE-Cys独特的表面结构和电荷分布使其具有很高的亲水性，有利于去除共存的盐和内源性血清肽。此外，本富集方法是并行处理的，从而节省了时间。使用这种方法，总共鉴定了从人血清蛋白中释放的47种独特的N-聚糖。47种N-连接聚糖的批内和批间变异系数分别为8.57% +/- 0.96%和9.22% +/-1.03%。这些结果表明，本方法适用于快速纯化人血清糖蛋白中的N-连接聚糖，并具有潜在的临床应用。
Expression levels of N-linked glycans derived from human serum glycoproteins have been shown to change during the progression of many diseases. Generally, N-glycans released from human serum proteins co-exist with endogenous serum peptides, salts, and other contaminants. Effective removal of these contaminants is essential to obtain the glycan profile of human serum proteins. Here, we developed a sample preparation method for mass spectrometry (MS) analysis of N-linked glycans derived from human serum glycoproteins based on a zwitterionic hydrophilic material named Click TE-Cys. The high hydrophilicity of Click TE-Cys, resulting from its unique surface structure and charge distribution, facilitated removal of co-existing salts and endogenous serum peptides. Furthermore, the present enrichment approach was handled in parallel, thus saving time. Using this method, a total of 47 unique N-glycans released from human serum proteins were identified. The intrabatch and interbatch coefficients of variation for the 47 N-linked glycans were 8.57% +/- 0.96% and 9.22% +/- 1.03%, respectively. These results demonstrate that the present method is suitable for fast purification of N-linked glycans derived from human serum glycoproteins, and has potential for clinical application.