title: Fc specific IgG glycosylation profiling by robust nano-reverse phase HPLC-MS using a sheath-flow ESI sprayer interface.
abstract: Biological activities of immunoglobulin G such as effector functions via Fc receptor interactions are influenced by Fc-linked N-glycans. Here we describe a fast, robust and sensitive nano-LC-ESI-MS method for detailed subclass specific analysis of IgG Fc N-glycosylation. A sheath-flow ESI sprayer was used as a sensitive zero dead volume plug-and-play interface for online MS coupling, generating a very constant spray and ionization over the entire LC gradient. The propionic acid containing sheath-liquid effectively suppressed TFA gas-phase ion-pairing, enabling the use of TFA containing mobile phases. The fixed position of the sheath-flow ESI sprayer, far away from the glass capillary inlet, reduced MS contamination as compared to conventional nano-ESI. The method was found to be suitable for fast and detailed subclass specific IgG Fc N-glycosylation profiling in human plasma. The obtained subclass specific IgG Fc N-glycosylation profiles were processed automatically using in house developed software tools. For each of the IgG subclasses the 8 major glycoforms showed an interday analytical variation below 5%. The method was used to profile the IgG Fc N-glycosylation of 26 women at several time points during pregnancy and after delivery, revealing pregnancy-associated changes in IgG galactosylation, sialylation and incidence of bisecting N-acetylglucosamine.
authors: Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.
authors: Maurice H J
authors: MH
authors: Selman
authors: Rico J E
authors: RJ
authors: Derks
authors: Albert
authors: A
authors: Bondt
authors: Magnus
authors: M
authors: Palmblad
authors: Bart
authors: B
authors: Schoenmaker
authors: Carolien A M
authors: CA
authors: Koeleman
authors: Fleur E
authors: FE
authors: van de Geijn
authors: Radboud J E M
authors: RJ
authors: Dolhain
authors: André M
authors: AM
authors: Deelder
authors: Manfred
authors: M
authors: Wuhrer
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005260:Female; D005690:Galactose / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism*; D007074:Immunoglobulin G / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D049590:Postpartum Period; D011247:Pregnancy; D012984:Software; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D006023:Glycoproteins
chemical_list: D000117:Acetylglucosamine; D005690:Galactose; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D019158:N-Acetylneuraminic Acid; D011134:Polysaccharides; C046179:glycosylated IgG; D006023:Glycoproteins