title: Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species.
abstract: The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. Since oligosaccharides play a significant role in many biological processes it is very important to understand their structure. The glycosylation is cell type specific as well as highly variable depending on the species producing the IgG. We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques.
authors.affiliation: GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland; Medical Biochemistry, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Barbara
authors.initials: B
authors.lastname: Adamczyk
authors.affiliation: GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland.
authors.forename: Tharmala
authors.initials: T
authors.lastname: Tharmalingam-Jaikaran
authors.affiliation: GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland.
authors.forename: Michael
authors.initials: M
authors.lastname: Schomberg
authors.affiliation: Horváth Laboratory for Bioseparation Sciences, Research Center for Molecular Medicine, University of Debrecen, 4032 Debrecen, Hungary.
authors.forename: Ákos
authors.initials: Á
authors.lastname: Szekrényes
authors.affiliation: GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland.
authors.forename: Ronan M
authors.initials: RM
authors.lastname: Kelly
authors.affiliation: Medical Biochemistry, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Niclas G
authors.initials: NG
authors.lastname: Karlsson
authors.affiliation: Horváth Laboratory for Bioseparation Sciences, Research Center for Molecular Medicine, University of Debrecen, 4032 Debrecen, Hungary; MTA-TKI Translational Glycomics Research Group, University of Pannonia, Veszprem, Hungary.
authors.forename: Andràs
authors.initials: A
authors.lastname: Guttman
authors.affiliation: GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland. Electronic address: pauline.rudd@nibrt.ie.
authors.forename: Pauline M
authors.initials: PM
authors.lastname: Rudd
mesh_terms: D000818:Animals; D002415:Cats; D002417:Cattle; D005591:Chemical Fractionation / Q000379:methods*; D004285:Dogs; D006031:Glycosylation; D006262:Health; D006736:Horses; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*
chemical_list: D007074:Immunoglobulin G; D009844:Oligosaccharides; D011134:Polysaccharides
keywords: CE-LIF; Glycan analysis; HILIC–UPLC; IgG; Protein glycosylation; RP–UPLC