Highly Efficient Enrichment of O-GalNAc Glycopeptides by Using Immobilized Metal Ion Affinity Chromatography
Highly Efficient Enrichment of O-GalNAc Glycopeptides by Using Immobilized Metal Ion Affinity Chromatography
复制标题
利用固定化金属离子亲和色谱法高效富集 O-GalNAc 糖肽
DOI：
10.1021/acs.analchem.0c05236
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发表时间：
2021
影响因子：
7.4
通讯作者：
Ye Mingliang
中科院分区：
文献类型：
--
作者：
Yue Xuyang;Qin Hongqiang;Chen Yao;Fang Zheng;Liu Luyao;Zhu He;Liu Xiaoyan;Zhou Jiahua;Tian Kailu;Qiao Xiaoqiang;Ye Mingliang
O-GalNAc糖基化的蛋白质组学分析对于筛选生物标志物和评估治疗反应是重要的。然而，由于目前可用的浓缩方法性能不佳，其分析仍面临挑战。本研究建立了一种基于Ti-IMAC（IV）材料的富集方法，该方法可以通过亲水性相互作用和亲和性相互作用富集完整的O-GalNAc糖肽。该方法仅从0.1 μL人血清中鉴定出近200种完整的O-GalNAc糖肽。这与HILIC方法的差异接近2倍。对O-GalNAc糖基化进行了深入分析，从7.2 μL人血清样品中鉴别出2093个完整糖肽。这是来自微量样品的最大的人血清O-GalNAc糖基化数据库。此外，通过对肝细胞癌（HCC）和对照血清样品的定量分析，确定了52个显著改变的完整O-GalNAc糖肽，表明这种富集方法在生物标志物发现中的潜在应用。
Proteomics analysis of O-GalNAc glycosylation is important for the screening of biomarkers and the assessment of therapeutic responses. However, its analysis still faces challenges due to the poor performance of currently available enrichment methods. In this study, an enrichment method was established on the basis of Ti-IMAC(IV) materials, which could enrich the intact O-GalNAc glycopeptides via both the hydrophilic interaction and affinity interaction. This method enabled nearly 200 intact O-GalNAc glycopeptides identified from only 0.1 μL of human serum. This was nearly 2-fold different from that of the HILIC method. An in-depth analysis of the O-GalNAc glycosylation was performed, and 2093 intact glycopeptides were identified from 7.2 μL of human serum samples. This is the largest O-GalNAc glycosylation database of human serum from a trace amount of sample. Furthermore, 52 significantly changed intact O-GalNAc glycopeptides were determined by the quantitative analysis of hepatocellular carcinoma (HCC) and control serum samples, indicating the potential applications of this enrichment method in biomarker discovery.