Center for Clinical Mass Spectrometry, College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu 215123, China.
Center for Clinical Mass Spectrometry, College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu 215123, China.
J Chromatogr A. 2024 Oct 25;1735:465318. doi: 10.1016/j.chroma.2024.465318. Epub 2024 Aug 31.
Protein glycosylation, one of the most important biologically relevant post-translational modifications for biomarker discovery, faces analytical challenges due to heterogeneous glycosite, diverse glycans, and mass spectrometry limitations. Glycopeptide enrichment by removing abundant hydrophobic peptides helps overcome some of these obstacles. Hydrophilic interaction liquid chromatography (HILIC), known for its selectivity, glycan separations, intact glycopeptide enrichment, and compatibility with mass spectrometry, has seen recent advancements in stationary phases like Amide-80, glycoHILIC, amino acids or peptides for improved HILIC-based glycopeptide analysis. Utilization of these materials can improve glycopeptide enrichment through solid-phase extraction and separation via high-performance liquid chromatography. Additionally, using glycopeptides themselves to modify HILIC stationary phases holds promise for improving selectivity and sensitivity in glycosylation analysis. Additionally, HILIC has capability to assess the information about glycosites and structural information of glycans. This review summarizes recent breakthroughs in HILIC stationary materials, highlighting their impact on glycopeptide analysis. Ongoing research on advanced materials continues to refine HILIC's performance, solidifying its value as a tool for exploring protein glycosylation.
蛋白质糖基化是生物相关性最强的翻译后修饰之一，对于生物标志物的发现具有重要意义，但由于糖基化位点的异质性、糖链的多样性以及质谱分析的限制，其分析存在挑战。通过去除丰富的疏水性肽来富集糖肽有助于克服这些障碍。亲水作用色谱（HILIC）因其选择性、糖链分离、完整糖肽的富集以及与质谱的兼容性而备受关注，近年来在固定相方面取得了进展，如酰胺-80、糖基 HILIC、氨基酸或肽，以提高基于 HILIC 的糖肽分析。这些材料的使用可以通过固相萃取和高效液相色谱分离来改善糖肽的富集。此外，利用糖肽本身来修饰 HILIC 固定相有望提高糖基化分析的选择性和灵敏度。此外，HILIC 还能够评估糖基化位点的信息和聚糖的结构信息。本综述总结了 HILIC 固定相材料的最新突破，强调了它们对糖肽分析的影响。对先进材料的持续研究不断改进 HILIC 的性能，巩固了其作为探索蛋白质糖基化工具的价值。
