Mass Spectrometry of Glycoproteins, Softcover reprint of the original 1st ed. 2013 Methods and Protocols Methods in Molecular Biology Series, Vol. 951

 Glycosylation is the most abundant post-translational modification of proteins.  Estimates vary widely, but a common assessment is that upwards of 50% of eukaryotic proteins are modified by some type of glycan. In Mass Spectrometry of Glycoproteins: Methods and Protocols, expert researchers in the field detail many of the methods that are now commonly used for glycoproteomics.  These methods and techniques include robust sample preparation techniques; advanced chromatographic strategies for improving dynamic range; state-of-the-art mass spectrometry instrumentation and associated ionization and fragmentation methods; and informatics tools used for identifying glycoproteins and characterizing the associated glycans. Written in the highly successful Methods in Molecular Biology? series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Introduction to Glycosylation and Mass Spectrometry.-Tandem Lectin Weak Affinity Chromatography (LWAC) for Glycoprotein Enrichment.-CSC Technology - Selective Labeling of Glycoproteins by Mild Oxidation to Phenotype Cells.-Use of Boronic Acid Nanoparticles in Glycoprotein Enrichment.-Incorporation of Unnatural Sugars for the Identification of Glycoproteins.-Characterization of Membrane-associated Glycoproteins using Lectin Affinity Chromatography and Mass Spectrometry.-Sialic Acid Capture-and-Release and LC-MSⁿ Analysis of Glycopeptides.-In-Solution Digestion of Glycoproteins for Glycopeptide-Based Mass Analysis.-Nano-HPLC-MS of Glycopeptides Obtained After Non-specific Proteolysis.-Glycopeptide Enrichment for MALDI-TOF Mass Spectrometry Analysis by Hydrophilic Interaction Liquid Chromatography Solid Phase Extraction (HILIC SPE).-Separation and Identification of Glycoforms by Capillary Electrophoresis with Electrospray Ionization Mass Spectrometric Detection.-Structural Separations by Ion Mobility-MS for Glycomics and Glycoproteomics.-Quantitative Analysis of Glycoprotein Glycans.-Stable Isotope Labeling of N-glycosylated Peptides by Enzymatic Deglycosylation for Mass Spectrometry-based Glycoproteomics.-Approaches for Site Mapping and Quantification of O-linked Glycopeptides.-Glycan Profiling: Label-free Analysis of Glycoproteins.-Introduction to Informatics in Glycoprotein Analysis.-Software Tools for Glycan Profiling.-Quantitative Characterization of Glycoproteins in Neurodegenerative Disorders using iTRAQ.-Quantitative Proteomic Analysis of N-linked Glycoproteins in Human Tear Fluid.-Elucidation of N-glycosites Within Human Plasma Glycoproteins for Cancer Biomarker Discovery.-Characterizing the Glycosylation State of Therapeutic RecombinantGlycoproteins.