title: Sialic Acid Derivatization for the Rapid Subclass- and Sialic Acid Linkage-Specific MALDI-TOF-MS Analysis of IgG Fc-Glycopeptides.
abstract: Matrix-assisted laser desorption/ionization (MALDI)-time-of-flight (TOF)-mass spectrometry (MS) is a highly suitable method for the rapid analysis of IgG glycopeptides, providing a wealth of structural information. A limitation of this approach is that it generates a bias when analyzing sialylated species due to the labile nature of sialic acid glycosidic linkages. One way to overcome this problem is by chemical derivatization of the sialic acids. The method presented here results in both the stabilization of the sialic acids, as well as the differentiation of α2,3- and α2,6-linked sialic acids by mass. Described in this chapter are the isolation of IgG from plasma or serum, tryptic digestion of the samples, derivatization, and finally MALDI-TOF-MS measurement and data analysis.
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333, ZA, Leiden, The Netherlands.
authors.forename: Noortje
authors.initials: N
authors.lastname: de Haan
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333, ZA, Leiden, The Netherlands.
authors.forename: Karli R
authors.initials: KR
authors.lastname: Reiding
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333, ZA, Leiden, The Netherlands. m.wuhrer@lumc.nl.|Division of BioAnalytical Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081, HV, Amsterdam, The Netherlands. m.wuhrer@lumc.nl.
authors.forename: Manfred
authors.initials: M
authors.lastname: Wuhrer
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D006020:Glycopeptides / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification; D006031:Glycosylation; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification; D012794:Sialic Acids / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D052616:Solid Phase Extraction / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D006020:Glycopeptides; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D012794:Sialic Acids
keywords: Biopharmaceuticals; Clinical cohorts; Cotton hydrophilic interaction liquid chromatography (HILIC)Matrix assisted laser desorption/ionization (MALDI); Fragment crystallizable (Fc)-glycosylation; Glycopeptide; Immunoglobulin G (IgG); Linkage-specific; Mass spectrometry (MS); N-acetylneuraminic acid; N-glycolylneuraminic acid; Sialic acid; Site-specific; Solid-phase extraction (SPE); Stabilization; Subclass-specific; Time-of-flight (TOF)