Department of General Surgery, Affiliated Union Hospital of Huazhong University of Science and Technology, Wuhan, Hubei Province, P. R. China.
J Sep Sci. 2011 Oct;34(19):2745-50. doi: 10.1002/jssc.201100427. Epub 2011 Sep 6.
Comprehensive glycoprotein characterization based on mass spectrometry (MS) is challenging because of low concentration of glycopeptides and suppression effect of abundant non-glycosylated peptides in MS. Therefore, it is vital to enrich glycopeptides before MS analysis. A new method was developed to selectively enrich glycopeptides from complex sample by coupling C18 fractionation with titanium dioxide (TiO(2)) enrichment. The new method allows to selectively enrich N-linked glycopeptides with various glycan forms and different sequence lengths. Compared with single TiO(2) method, the established method demonstrated higher glycopeptide selectivity and higher glycosylation heterogeneity coverage. Further application of this method to mixture of non-glycosylated protein and glycoprotein digests at different levels reveals the feasibility of enrichment of tryptic glycopeptides from simple proteomics samples.
基于质谱（MS）的全面糖蛋白特征分析具有挑战性，因为糖肽浓度低，且 MS 中丰富的非糖基化肽会产生抑制效应。因此，在 MS 分析之前对糖肽进行富集至关重要。本研究开发了一种新方法，通过 C18 分级与二氧化钛（TiO2）富集相结合，从复杂样品中选择性富集糖肽。该新方法可选择性富集具有各种聚糖形式和不同序列长度的 N 连接糖肽。与单一 TiO2 方法相比，所建立的方法显示出更高的糖肽选择性和更高的糖基化异质性覆盖率。该方法进一步应用于不同水平的非糖基化蛋白和糖蛋白消化物的混合物，表明从简单蛋白质组学样品中富集胰蛋白酶糖肽是可行的。
