title: CSC technology: selective labeling of glycoproteins by mild oxidation to phenotype cells.
abstract: Cell surface glycoproteins represent important markers for the phenotyping of healthy and malignantly transformed cells. The mass spectrometry-based cell surface capturing (CSC) technology allows for extensive multiplexed identification and relative quantification of glycoproteins expressed on the cell surface at a given point in time. CSC technology is based on the mild oxidation of glycans from cell surface proteins on living cells. Oxidized glycans are tagged with a bifunctional linker molecule and glycopeptides are subsequently enriched by affinity chromatography. Here, we describe a step-by-step protocol of the CSC technology, which not only enables the identification of cell surface glycoproteins, but also the concurrent determination of protein N-glycosylation sites.
authors.affiliation: Institute of Molecular Systems Biology, Swiss Federal Institute of Technology (ETH) Zurich, Zurich, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hofmann
authors.forename: Damaris
authors.initials: D
authors.lastname: Bausch-Fluck
authors.forename: Bernd
authors.initials: B
authors.lastname: Wollscheid
mesh_terms: D000595:Amino Acid Sequence; D019921:Biotinylation; D002462:Cell Membrane / Q000378:metabolism; D002853:Chromatography, Liquid; D006020:Glycopeptides / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D010641:Phenotype; D059748:Proteolysis; D040901:Proteomics; D012492:Salts / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D006020:Glycopeptides; D008562:Membrane Glycoproteins; D012492:Salts