title: Comparison of procainamide and 2-aminobenzamide labeling for profiling and identification of glycans by liquid chromatography with fluorescence detection coupled to electrospray ionization-mass spectrometry.
abstract: One of the most widely used methods for glycan analysis is fluorescent labeling of released glycans followed by hydrophilic interaction chromatography-(ultra-)high-performance liquid chromatography [HILIC-(U)HPLC]. Here, we compare the data obtained by (U)HPLC-fluorescence (FLR) coupled to electrospray ionization-mass spectrometry (ESI-MS) for procainamide and 2-aminobenzamide (2-AB)-labeled N-glycans released from human immunoglobulin G (IgG). Fluorescence profiles from procainamide show comparable chromatographic separation to those obtained for 2-AB but gave higher fluorescence intensity as well as significantly improved ESI efficiency (up to 30 times that of 2-AB). Thus, labeling with procainamide increases the ability to identify minor glycan species that may have significant biological activity.
authors.affiliation: Ludger Ltd., Culham Science Centre, Oxfordshire OX14 3EB, UK. Electronic address: rad.kozak@ludger.com.
authors.forename: Radoslaw P
authors.initials: RP
authors.lastname: Kozak
authors.affiliation: Ludger Ltd., Culham Science Centre, Oxfordshire OX14 3EB, UK.
authors.forename: Concepcion Badia
authors.initials: CB
authors.lastname: Tortosa
authors.affiliation: Ludger Ltd., Culham Science Centre, Oxfordshire OX14 3EB, UK.
authors.forename: Daryl L
authors.initials: DL
authors.lastname: Fernandes
authors.affiliation: Ludger Ltd., Culham Science Centre, Oxfordshire OX14 3EB, UK.
authors.forename: Daniel I R
authors.initials: DI
authors.lastname: Spencer
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry*; D011342:Procainamide / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013194:Staining and Labeling; D062367:ortho-Aminobenzoates / Q000737:chemistry*
chemical_list: D005456:Fluorescent Dyes; D011134:Polysaccharides; D062367:ortho-Aminobenzoates; D011342:Procainamide; C000219:anthranilamide
keywords: ESI–MS; Fluorescent labeling; Glycan derivatization; HILIC–(U)HPLC; Procainamide