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使用纤维素亲水相互作用/反相阶段Tips快速富集糖肽

Rapid Glycopeptide Enrichment Using Cellulose Hydrophilic Interaction/Reversed-Phase StageTips.

作者信息

Ohta Yuki, Kameda Kotaro, Matsumoto Mei, Kawasaki Nana

机构信息

Laboratory of Biopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City University.

Laboratory of Biopharmaceutical and Regenerative Sciences, Medical Life Science, Division of Sciences, International College of Arts and Sciences, Yokohama City University.

出版信息

Mass Spectrom (Tokyo). 2017;6(1):A0061. doi: 10.5702/massspectrometry.A0061. Epub 2017 Aug 23.

DOI:10.5702/massspectrometry.A0061
PMID:28852604
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5570719/
Abstract

Because the ionization efficiency for glycopeptides is lower than that of peptides in electrospray ionization, it is frequently necessary to enrich them prior to their analysis using liquid chromatography coupled with tandem mass spectrometry. Although some methods for selectively enriching glycopeptides (, lectin, agarose, and cellulose methods) have been reported, they are time-consuming (procedures that require several hours) and may not be applicable to submicrogram-sized samples. Here, we report on a rapid, simple method for enriching glycopeptides in small sample amounts using cellulose hydrophilic interaction (cellulose HILIC)/reversed-phase (RP) stop-and-go extraction tips (StageTips). Using the cellulose HILIC/RP StageTips, glycopeptide-selective enrichment can be achieved at the nanogram level within a few minutes.

摘要

由于在电喷雾电离中糖肽的电离效率低于肽，因此在使用液相色谱-串联质谱进行分析之前，通常需要对其进行富集。尽管已经报道了一些选择性富集糖肽的方法（如凝集素法、琼脂糖法和纤维素法），但这些方法耗时较长（需要几个小时的操作过程），并且可能不适用于亚微克级别的样品。在此，我们报告一种使用纤维素亲水相互作用（纤维素HILIC）/反相（RP）停停走走萃取尖管（StageTips）对少量样品中的糖肽进行快速、简便富集的方法。使用纤维素HILIC/RP StageTips，可以在几分钟内实现纳克级别的糖肽选择性富集。