Glycomic Analysis of Glycans Released from Glycoproteins Using Chemical Immobilization and Mass Spectrometry. Issue 3 (9th September 2014)

• Record Type: Journal Article
• Authors: Yang, Shuang; Zhang, Hui
• Editors: Mahal, Lara; Romesberg, Floyd; Shah, Kavita; Shamu, Caroline; Strano, Michael S.; Thomas, Craig
• Is Part Of: Current protocols in chemical biology. Volume 6:Issue 3(2014)
• Page Start: 191
• Page End: 208
• Publication Date: 2014-09-09
• Subjects: solid‐phase -- protein immobilization -- glycan derivatization -- chemoenzymatic processes -- MALDI -- GIG -- mass spectrometry -- glycomics; Biochemistry -- Laboratory manuals; Chemical Phenomena; Biochemistry; Periodicals; Laboratory manuals; 572.078
• DOI: 10.1002/9780470559277.ch140085
• Languages: English
• ISSNs: 2160-4762

Abstract

Abstract: Protein glycosylation is one of most common protein modifications and is involved in many biological activities. N‐linked and O‐linked glycosylation not only represent abundant glycan modifications, but also are structurally diverse. Mass spectrometry has emerged as a major method for glycomic analysis. However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. In this protocol, we describe a novel and high‐throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid‐phase. Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. Glycans are then released from the solid support for analysis by mass spectrometry. The procedures outlined are robust and useful for high‐throughput glycomic analysis from complex biological or clinical samples. Curr. Protoc. Chem. Biol . 6:191‐208 © 2014 by John Wiley & Sons, Inc.