title: N-linked global glycan profiling by nanoLC mass spectrometry.
abstract: A method is detailed for the global profiling of underivatized N-linked glycans that are derived from complex protein mixtures. The method consists of five main steps that include the following: (1) protein denaturation; (2) enzymatic digestion; (3) solid phase extraction; (4) nanoLC MS analysis; and (5) data interpretation. Materials, methods, and algorithms for the identification of both glycan composition and structure are summarized. In addition, potential problems and their resolutions are addressed.
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, WA, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Bereman
authors.forename: David C
authors.initials: DC
authors.lastname: Muddiman
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D001216:Asparagine; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D006023:Glycoproteins / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D008358:Mannose / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D009584:Nitrogen / Q000737:chemistry*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011489:Protein Denaturation; D052616:Solid Phase Extraction; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry
chemical_list: D001798:Blood Proteins; D006023:Glycoproteins; D011134:Polysaccharides; D001216:Asparagine; D006026:Glycoside Hydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D009584:Nitrogen; D008358:Mannose; D000117:Acetylglucosamine