Kong Siyuan, Zhang Quanqing, Yang Lujie, Huang Yuanyu, Liu Mingqi, Yan Guoquan, Zhao Huanhuan, Wu Mengxi, Zhang Xiangmin, Yang Pengyuan, Cao Weiqian
The Fifth People's Hospital, Fudan University, and the Shanghai Key Laboratory of Medical Epigenetics, the International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.
NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai 200032, China.
Anal Chem. 2021 May 4;93(17):6682-6691. doi: 10.1021/acs.analchem.0c05482. Epub 2021 Apr 20.
The heterogeneity and low abundance of protein glycosylation present challenging barriers to the analysis of intact glycopeptides, which is key to comprehensively understanding the role of glycosylation in an organism. Efficient and specific enrichment of intact glycopeptides could help greatly with this problem. Here, we propose a new enrichment strategy using a boronic acid (BA)-functionalized mesoporous graphene-silica composite (denoted as GO@mSiO-GLYMO-APB) for isolating intact glycopeptides from complex biological samples. The merits of this composite, including high surface area and synergistic effect from size exclusion functionality of mesoporous material, hydrophilic interaction of silica, and the reversible covalent binding with BA, enable the effective and specific enrichment of both intact N- and O-glycopeptides. The results from the enrichment performance of the strategy evaluated by standard glycoproteins and the application to global N- and O-glycosylation analyses in human serum indicate the robustness and potential of the strategy for intact glycopeptide analysis.
蛋白质糖基化的异质性和低丰度给完整糖肽的分析带来了具有挑战性的障碍，而完整糖肽分析是全面了解糖基化在生物体中作用的关键。高效且特异性地富集完整糖肽有助于极大地解决这一问题。在此，我们提出一种新的富集策略，即使用硼酸（BA）功能化的介孔石墨烯-二氧化硅复合材料（记为GO@mSiO-GLYMO-APB）从复杂生物样品中分离完整糖肽。这种复合材料的优点，包括高比表面积以及介孔材料的尺寸排阻功能、二氧化硅的亲水相互作用和与BA的可逆共价结合所产生的协同效应，能够有效且特异性地富集完整的N-糖肽和O-糖肽。通过标准糖蛋白评估该策略的富集性能以及将其应用于人类血清中全局N-糖基化和O-糖基化分析的结果，表明了该策略在完整糖肽分析方面的稳健性和潜力。
