Wang Heping, Jiao Fenglong, Gao Fangyuan, Huang Junjie, Zhao Yan, Shen Yehua, Zhang Yangjun, Qian Xiaohong
State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing 102206, China.
J Mater Chem B. 2017 Jun 14;5(22):4052-4059. doi: 10.1039/c7tb00700k. Epub 2017 May 16.
Protein glycosylations play important roles in various biological processes and in the disease progression of organisms. The development of specific enrichment materials and strategies before mass spectrometric analysis was a prerequisite to glycoproteomic analysis due to the difficulty caused by substoichiometric levels of glycoproteins. In this work, novel magnetic covalent organic frameworks (denoted as FeO@TpPa-1) were first developed using only a two-step solvothermal reaction and then applied in the hydrophilic enrichment of glycopeptides. Sea urchin-type composites with super-paramagnetic properties were constructed by in situ growth of TpPa-1 covalent organic frameworks on the surface of magnetic nanoparticles. A total of 37 and 22 glycopeptides could be easily detected from IgG and HRP digests, respectively, by hydrophilic enrichment with the newly developed materials. An ultralow detection limit (28 fmol), satisfactory selectivity and high recovery could be achieved using FeO@TpPa-1. The material's excellent enrichment performance was also demonstrated using glycopeptide analysis in real complex samples. Within three independent replicates, 228 glycopeptides corresponding to 114 glycoproteins could be detected from human serum digests, which is better than the performance obtained by commercial HILIC materials. The results suggest that covalent organic frameworks show potential for application in glycoproteomic studies.
蛋白质糖基化在各种生物过程和生物体疾病进展中发挥着重要作用。由于糖蛋白亚化学计量水平所带来的困难，在质谱分析之前开发特定的富集材料和策略是糖蛋白质组学分析的先决条件。在这项工作中，仅通过两步溶剂热反应首次开发了新型磁性共价有机框架（表示为FeO@TpPa-1），然后将其应用于糖肽的亲水富集。通过在磁性纳米颗粒表面原位生长TpPa-1共价有机框架构建了具有超顺磁性的海胆型复合材料。使用新开发的材料进行亲水富集，分别从IgG和HRP消化物中可以轻松检测到总共37种和22种糖肽。使用FeO@TpPa-1可以实现超低检测限（28 fmol）、令人满意的选择性和高回收率。在实际复杂样品中的糖肽分析也证明了该材料优异的富集性能。在三个独立重复实验中，从人血清消化物中可以检测到对应于114种糖蛋白的228种糖肽，这比商业亲水作用色谱材料获得的性能更好。结果表明，共价有机框架在糖蛋白质组学研究中具有应用潜力。
