Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification
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- 作者：Marco R. Bladergroen ; Karli R. Reiding ; Agnes L. Hipgrave Ederveen ; Gerda C.M. Vreeker ; Florent Clerc ; Stephanie Holst ; Albert Bondt ; Manfred Wuhrer ; Yuri E.M. van der Burgt
- 刊名：Journal of Proteome Research
- 出版年：2015
- 出版时间：September 4, 2015
- 年：2015
- 卷：14
- 期：9
- 页码：4080-4086
- 全文大小：444K
- ISSN：1535-3907
文摘
Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. Previously, we have developed a robust, high-throughput MALDI-TOF鈥揗S method for the comprehensive profiling of human plasma N-glycans. In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of 伪2,6-linked sialic acid residues with parallel lactone formation of 伪2,3-linked sialic acids. In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing.