title: Glycomic analysis using glycoprotein immobilization for glycan extraction.
abstract: Glycosylation is one of the most common protein modifications and is involved in many functions of glycoproteins. Investigating aberrant protein glycosylation associated with diseases is useful in improving disease diagnostics. Due to the nontemplate nature of glycan biosynthesis, the glycans attached to glycoproteins are enormously complex; thus, a method for comprehensive analysis of glycans from biological or clinical samples is needed. Here, we describe a novel method for glycomic analysis using glycoprotein immobilization for glycan extraction (GIG). Proteins or peptides from complex samples were first immobilized on solid support, and other nonconjugated molecules were removed. Glycans were enzymatically or chemically modified on solid phase before releasing from glycoproteins/glycopeptides for mass spectrometry analysis. The method was applied to the glycomic analysis of both N- and O-glycans.
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, MD 21231, USA.
authors.forename: Shuang
authors.initials: S
authors.lastname: Yang
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Punit
authors.initials: P
authors.lastname: Shah
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
mesh_terms: D054794:Glycomics; D006020:Glycopeptides / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D009077:Mucins / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis* / Q000302:isolation & purification* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006020:Glycopeptides; D006023:Glycoproteins; D009077:Mucins; D011134:Polysaccharides