Novel boronate material affords efficient enrichment of glycopeptides by synergized hydrophilic and affinity interactions
Novel boronate material affords efficient enrichment of glycopeptides by synergized hydrophilic and affinity interactions
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新型硼酸盐材料通过协同的亲水性和亲和力相互作用提供有效的糖肽富集
DOI：
10.1007/s00216-016-0044-5
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发表时间：
2017-01-01
影响因子：
4.3
通讯作者：
Zhang, Bo
中科院分区：
文献类型：
--
作者：
Chen, Jianying;Li, Xiaohu;Zhang, Bo
开发富集糖肽的新材料是全面分析糖蛋白组的关键，糖蛋白组与多种重大疾病和生物标志物的发现密切相关。我们通过硫醇-烯点击化学方法合成了与SiO2微球结合的苯基硼酸（PBA）（该材料记为点击PBA），并用其在亲水相互作用色谱模式下分离含顺式二醇的分子和富集糖肽。通过元素分析、X射线光电子能谱、N2吸附-脱附等温线和高分辨率扫描电子显微镜证实了点击PBA的成功制备。 Click PBA 在碱性、有机溶液含量较高的条件下比在酸性、有机溶液含量较高或碱性水溶液条件下对糖肽表现出更强的保留。 Click PBA 对中性和酸性糖肽均表现出高选择性，可抵抗 100 摩尔倍牛血清白蛋白消化物的干扰。高富集效率可归因于亲和相互作用和亲水相互作用的协同效应。将 Click PBA 应用到 1 µL 人血清中，从 71 种糖蛋白中鉴定出了 101 个独特的糖基化位点。这种材料将有助于全面的糖蛋白质组分析。
Development of novel materials for enrichment of glycopeptides is the key to a comprehensive analysis of the glycoproteome, which is closely related to several major diseases and biomarker findings. We synthesized phenylboronic acid (PBA) bound to SiO2microspheres by a thiol–ene click chemistry method (this material was denoted as click PBA) and used it to separatecis-diol-containing molecules and enrich glycopeptides in hydrophilic interaction chromatography mode. Successful preparation of click PBA was confirmed by elemental analysis, X-ray photoelectron spectroscopy, N2adsorption–desorption isotherms, and high-resolution scanning electron microscopy. Click PBA showed stronger retention towards glycopeptides under alkaline, higher content of organic solution conditions than under acidic, higher content of organic solution or alkaline aqueous solution conditions. Click PBA exhibited high selectivity for both neutral and acidic glycopeptides, which could resist interference from 100 molar fold of bovine serum albumin digests. The high enrichment efficiency can be ascribed to the synergetic effects of affinity interaction and hydrophilic interaction. The application of click PBA to 1 μL human serum resulted in the identification of 101 unique glycosylation sites from 71 glycoproteins. Such material will facilitate comprehensive glycoproteome analysis.