Identification and quantification of glycans from the physico-chemical viewpoint using analytical strategies, glycan release methods, and small-scale purification protocols are essential to address the difficulties of structural analysis in glycobiology at different degrees of complexity. This chapter addresses the contemporary analytical methodologies used in molecular profiling and mass spectroscopy (MS)-based structural elucidation of N-glycans cleaved from glycoproteins. The tasks and analytical attributes of these determinations in different aspects of glycobiology vary in terms of difficulty, complexity of materials, and the amounts of available glycoproteins. The glycomic analysis of asparagine-linked sugars is viewed as a more facile target of investigation than the analysis of O-glycans. This situation is largely due to the availability of endoglycosidase enzymes and uniform core structure with biosynthetic attributes. An array of new instrumental approaches and MS tools addresses the key structural and functional aspects of glycoconjugates in several ways. The new types of mass analyzers and tandem MS techniques work at unprecedented sensitivity and mass resolution to determine sugar sequences, chain branching, linkages, and additional aspects of the oligosaccharide structure. The approaches are primarily based on chemical/analytical measurements such as MS and laser-induced fluorescence detection following the use of a capillary separation technique. Recent advances in the use of MS tandem techniques are discussed in detail, reviewing important research avenues and potential directions.
|Title of host publication||Handbook of Glycomics|
|Number of pages||43|
|State||Published - 2010|