Key Laboratory of Advanced Mass Spectrometry and Molecular Analysis of Zhejiang Province, School of Materials Science and Chemical Engineering, Institute of Mass Spectrometry, Ningbo University, Ningbo, China.
Institute of Drug Discovery Technology, Ningbo University, Ningbo, China.
Rapid Commun Mass Spectrom. 2022 Dec 30;36(24):e9405. doi: 10.1002/rcm.9405.
Glycosylation of proteins is one of the most significant and complex post-translational modifications, and N-glycosylation plays a crucial role in life activities. Mass spectrometry (MS) has been a powerful technique in the analysis of protein glycosylation. However, the direct detection of glycoproteins in biological samples based on MS still suffers from huge challenges. Therefore, enrichment and purification of samples before MS analysis is an essential prerequisite.
Hydrophilic interaction liquid chromatography (HILIC) has significantly developed for selective enrichment of glycopeptides due to its simple operation process and unbiased enrichment. Herein, hydrophilic, dual amino acid-functionalized zinc sulfide quantum dots (ZnS QDs) were prepared to enrich glycopeptides using an easy procedure. The enriched glycopeptides were detected using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).
The obtained material exhibited high selectivity (1:2000), low detection limit (0.1 fmol/μl), good repeatability (10 times), and excellent recovery (89.8%) in glycopeptide enrichment. In the actual application in biological samples, 71 N-glycopeptides and 161 N-glycopeptides were detected from human saliva and serum, respectively.
ZnS-Au-GC was successfully prepared using an easy method. The results showed that the obtained material exhibited excellent performance in glycopeptide enrichment. Furthermore, it had showed great potential for glycopeptide enrichment in complex biological samples.
蛋白质糖基化是最重要、最复杂的翻译后修饰之一，而 N-糖基化在生命活动中起着至关重要的作用。质谱（MS）技术已成为分析蛋白质糖基化的有力工具。然而，基于 MS 直接检测生物样品中的糖蛋白仍然面临巨大挑战。因此，在 MS 分析之前对样品进行富集和纯化是必不可少的前提条件。
亲水作用色谱（HILIC）由于其操作过程简单且具有非偏性的富集特点，已极大地发展用于选择性富集糖肽。在此，我们采用简单的方法制备了亲水性、双氨基酸功能化的硫化锌量子点（ZnS QDs），用于富集糖肽。采用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）检测富集的糖肽。
所获得的材料在糖肽富集方面表现出高选择性（1:2000）、低检测限（0.1 fmol/μl）、良好的重复性（10 次）和优异的回收率（89.8%）。在实际生物样品中的应用中，分别从人唾液和血清中检测到 71 个 N-糖肽和 161 个 N-糖肽。
采用简单的方法成功制备了 ZnS-Au-GC。结果表明，所获得的材料在糖肽富集方面表现出优异的性能。此外，它在复杂生物样品中的糖肽富集方面具有很大的潜力。
