A thiazolidine formation-based approach for ultrafast and highly efficient solid-phase extraction of N-Glycoproteome
A thiazolidine formation-based approach for ultrafast and highly efficient solid-phase extraction of N-Glycoproteome
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DOI：
10.1016/j.aca.2019.12.001
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发表时间：
2020-03-01
影响因子：
6.2
通讯作者：
Lu, Haojie
中科院分区：
文献类型：
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作者：
Cai, Yan;Zhang, Ying;Lu, Haojie
对于基于质谱（MS）的n -糖蛋白组学，在MS分析之前选择性富集n -糖肽是降低样品复杂性的关键步骤。基于共价偶联的富集由于其无偏性和高度特异性的特点而日益成为一种有吸引力的策略。然而，目前大多数用于富集n -糖肽的共价偶联反应仍受偶联时间长和偶联条件苛刻的限制。在此，我们开发了一种基于噻唑烷形成的方法，用于超快速高效地提取n-糖蛋白组。利用易合成的cys端磁性纳米粒子，通过噻唑烷的形成，糖肽上的氧化聚糖部分可以被磁性纳米粒子表面的β氨基巯基选择性地捕获。在温和条件下，无需添加有毒催化剂或破坏样品的还原剂，可在30 min内完成偶联。此外，在灵敏度（低fmol水平）、选择性（以1:100摩尔比从糖肽和非糖肽的混合物中提取n-糖肽）和重复性(cv
For mass spectrometry (MS)-based N-glycoproteomics, selective enrichment of N-glycopeptides prior to MS analysis is a crucial step to reduce sample complexity. Enrichment based on covalent coupling is as an increasingly attractive strategy due to the unbiased and highly specific features. However, most of current covalent coupling reactions for N-glycopeptides enrichment are still limited by long coupling time and harsh coupling conditions. Herein, we developed a thiazolidine formation-based approach for ultrafast and highly efficient solid-phase extraction of N-Glycoproteome. With the use of facile synthesis of Cys-terminated magnetic nanoparticles, the oxidized glycan moieties on glycopeptides could be selectively captured by the beta-amino thiols groups on the surface of magnetic nanoparticles through thiazolidine formation. The coupling could be achieved within 30 min under mild condition, eliminating the addition of toxic catalyst or sample-destroying reducing agent. Also, the great enrichment performance for N-glycopeptides were obtained in terms of sensitivity (low fmol levels), selectivity (extracting N-glycopeptides from the mixture of glycopeptides and non-glycopeptides at a 1:100 molar ratio) and reproducibility (CVs