Highly efficient enrichment of N-linked glycopeptides using a hydrophilic covalent-organic framework
Highly efficient enrichment of N-linked glycopeptides using a hydrophilic covalent-organic framework
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使用亲水性共价有机框架高效富集 N-连接糖肽
DOI：
10.1039/c7an01027c
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发表时间：
2017-09-07
期刊：
影响因子：
4.2
通讯作者：
Zhang, Xin-Xiang
中科院分区：
文献类型：
--
作者：
Ma, Yu-Fang;Yuan, Fang;Zhang, Xin-Xiang
糖肽的富集在糖蛋白质组学中起着重要的作用。本文首次将1，3，5-三甲酰间苯三酚与对苯二胺通过Schiff碱反应合成的共价有机骨架TpPa-1作为亲水性多孔材料用于N-连接糖肽的富集。该材料可有效地消除非糖肽的干扰，有利于糖肽的质谱检测。通过从人IgG的胰蛋白酶消化物中捕获N-连接的糖肽，我们的方法被证明在飞摩尔水平上具有高灵敏度。即使当非糖肽被浓缩1000倍时，它也显示出对糖肽的上级选择性。由于共价键较强，该材料具有良好的稳定性，可重复使用10次以上。超低的质量密度和丰富的结合位点也为它提供了高的结合容量（178 mg g（-1），IgG/TpPa（-1））。此外，N-连接的糖肽很容易富集这种材料从只有10 μ L的人血清，这表明其在复杂的生物样品的预处理的潜力。
The enrichment of glycopeptides plays an important role in glycoproteomics. In this paper, a covalent-organic framework called TpPa-1, synthesized by the Schiff base reaction of 1,3,5-triformylphloroglucinol and paraphenylenediamine, was first successfully utilized as a hydrophilic porous material for N-linked glycopeptide enrichment. Using this material, interference from non-glycopeptides could be efficiently eliminated, which facilitated the mass spectrometry detection of glycopeptides. By capturing N-linked glycopeptides from tryptic digests of human IgG, our method was proved to have high sensitivity at the femtomole level. And it showed superior selectivity for glycopeptides even when non-glycopeptides were 1000 times more concentrated. Due to the strong covalent bonds, this material possessed good stability and could be repeatedly used for at least 10 times. The ultra-low mass density and abundant binding sites also provided it with high binding capacity (178 mg g(-1), IgG/TpPa(-1)). Moreover, N-linked glycopeptides were easily enriched by this material from only 10 mu L human serum, which demonstrated its potential in pretreatment of complex biological samples.