Facile synthesis of carboxymethyl-beta-cyclodextrin conjugated magnetic nanoparticles for selective enrichment of glycopeptides
Facile synthesis of carboxymethyl-beta-cyclodextrin conjugated magnetic nanoparticles for selective enrichment of glycopeptides
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轻松合成羧甲基-β-环糊精共轭磁性纳米粒子用于选择性富集糖肽
DOI：
10.1002/rcm.7626
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发表时间：
2016
影响因子：
2
通讯作者：
Ruijun Tian
中科院分区：
文献类型：
--
作者：
Peipei Song;Peiwu Huang;Tengjun Huang;Hua Li;Wendong Chen;Lin Lin;Shun Feng;Ruijun Tian
在质谱（MS）分析之前，糖肽的选择性富集是必不可少的，因为在复杂样品中修饰的糖肽丰度低，在MS电离和检测过程中由非糖基化肽的共同存在引起的离子抑制作用等。在不同的富集方法中，基于亲水性相互作用液相色谱（HILIC）的磁分离因其对糖肽的富集效率高、选择性好而成为近年来最受欢迎的富集方法之一。方法采用碳二亚胺活化法制备新型羧甲基- β -环糊精（CMCD）修饰磁性纳米颗粒（MNPs）。CMCD通过碳二亚胺与MNPs表面的-OH基团共价结合，该方法具有相互作用稳定、节省时间、操作简单等优点，是一种快速高效的糖肽富集方法。结果制备的吸附剂平均直径为15 nm，对外加磁场有较强的磁响应。热重分析结果表明，结合CMCD的含量为3 wt%。在fe3o4核上共轭的CMCD外层具有较高的表面亲水性。在对复杂小鼠肝脏样本的分析中，成功鉴定了494个糖基化蛋白对应的666个独特的N -糖基化位点。结论：该研究证明了一种易于使用的基于CMCD修饰的MNPs的方法，具有高选择性和高容量，可从复杂生物样品中富集低丰度的糖肽。版权所有©2016 John Wiley & Sons， Ltd。
RationaleSelective enrichment of glycopeptides prior to mass spectrometry (MS) analysis is essential due to the low abundance of the modified glycopeptides in complex samples, ion suppression effects during MS ionization and detection caused by the co‐presence of non‐glycosylated peptides, etc. Among different enrichment approaches, hydrophilic interaction liquid chromatography (HILIC)‐based magnetic separation has become one of the most popular methods in recent years, due to its high efficiency and selectivity for glycopeptide enrichment.MethodsHerein, novel carboxymethyl‐β‐cyclodextrin (CMCD)‐modified magnetic nanoparticles (MNPs) were synthesized via a carbodiimide activation method. CMCD was covalently bonded with the –OH group on the surface of MNPs through carbodiimide, and the proposed procedure provides a rapid and efficient alternative for glycopeptide enrichment due to its stable interaction, time‐saving, and easy operation.ResultsThe prepared absorbents with a mean diameter of 15 nm demonstrated a strong magnetic response to an externally applied magnetic field. The results of thermogravimetric analysis showed the content of bound CMCD was 3 wt%. The outer CMCD layer conjugated on the Fe3O4core showed high hydrophilic surface property. In the analysis of a complex mouse liver sample, a total of 666 unique N‐glycosylation sites corresponding to 494 glycosylated proteins were identified successfully.ConclusionsThe study demonstrated an easy‐to‐use CMCD‐modified MNPs‐based approach with high selectivity and high capacity in the enrichment of low‐abundance glycopeptides from complex biological samples. Copyright © 2016 John Wiley & Sons, Ltd.