title: Negative-Ion Electrospray Tandem Mass Spectrometry and Microarray Analyses of Developmentally Regulated Antigens Based on Type 1 and Type 2 Backbone Sequences.
abstract: Type 1 (Galβ1-3GlcNAc) and type 2 (Galβ1-4GlcNAc) sequences are constituents of the backbones of a large family of glycans of glycoproteins and glycolipids whose branching and peripheral substitutions are developmentally regulated. It is highly desirable to have microsequencing methods that can be used to precisely identify and monitor these oligosaccharide sequences with high sensitivity. Negative-ion electrospray tandem mass spectrometry with collision-induced dissociation has been used for characterization of branching points, peripheral substitutions, and partial assignment of linkages in reducing oligosaccharides. We now extend this method to characterizing entire sequences of linear type 1 and type 2 chain-based glycans, focusing on the type 1 and type 2 units in the internal regions including the linkages connecting type 1 and type 2 disaccharide units. We apply the principles to sequence analysis of closely related isomeric oligosaccharides and demonstrate by microarray analyses distinct binding activities of antibodies and a lectin toward various combinations of type 1 and 2 units joined by 1,3- and 1,6-linkages. These sequence-specific carbohydrate-binding proteins are in turn valuable tools for detecting and distinguishing the type 1 and type 2-based developmentally regulated glycan sequences.
authors.affiliation: Glycosciences Laboratory, Department of Medicine, Imperial College London , Hammersmith Campus, London W12 0NN, U.K.
authors.forename: Chao
authors.initials: C
authors.lastname: Gao
authors.affiliation: Glycosciences Laboratory, Department of Medicine, Imperial College London , Hammersmith Campus, London W12 0NN, U.K.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Glycosciences Laboratory, Department of Medicine, Imperial College London , Hammersmith Campus, London W12 0NN, U.K.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Glycosciences Laboratory, Department of Medicine, Imperial College London , Hammersmith Campus, London W12 0NN, U.K.
authors.forename: Ten
authors.initials: T
authors.lastname: Feizi
authors.affiliation: Glycosciences Laboratory, Department of Medicine, Imperial College London , Hammersmith Campus, London W12 0NN, U.K.
authors.forename: Wengang
authors.initials: W
authors.lastname: Chai
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D000941:Antigens / Q000032:analysis*; D002236:Carbohydrate Conformation; D006017:Glycolipids / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D037121:Plant Lectins / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D040081:Protein Array Analysis; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D006017:Glycolipids; D006023:Glycoproteins; D037121:Plant Lectins; D011134:Polysaccharides; C017707:Ricinus communis agglutinin-1