title: Chemoenzymatic method for glycomics: Isolation, identification, and quantitation.
abstract: Over the past decade, considerable progress has been made with respect to the analytical methods for analysis of glycans from biological sources. Regardless of the specific methods that are used, glycan analysis includes isolation, identification, and quantitation. Derivatization is indispensable to increase their identification. Derivatization of glycans can be performed by permethylation or carbodiimide coupling/esterification. By introducing a fluorophore or chromophore at their reducing end, glycans can be separated by electrophoresis or chromatography. The fluorogenically labeled glycans can be quantitated using fluorescent detection. The recently developed approaches using solid-phase such as glycoprotein immobilization for glycan extraction and on-tissue glycan mass spectrometry imaging demonstrate advantages over methods performed in solution. Derivatization of sialic acids is favorably implemented on the solid support using carbodiimide coupling, and the released glycans can be further modified at the reducing end or permethylated for quantitative analysis. In this review, methods for glycan isolation, identification, and quantitation are discussed.
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, USA.
authors.forename: Shuang
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, USA.
authors.forename: Abigail
authors.initials: A
authors.lastname: Rubin
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, USA.
authors.forename: Shadi Toghi
authors.initials: ST
authors.lastname: Eshghi
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry; D000818:Animals; D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid; D054794:Glycomics / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry; D011134:Polysaccharides / Q000302:isolation & purification*; D011499:Protein Processing, Post-Translational; D017421:Sequence Analysis
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides; D000581:Amidohydrolases
keywords: Glycomics; Glycoproteomics; Isobaric labeling; Mass spectrometry; iGIG