title: On-line sheathless capillary electrophoresis/nanoelectrospray ionization-tandem mass spectrometry for the analysis of glycosaminoglycan oligosaccharides.
abstract: A novel approach in glycosaminoglycomics, based on sheathless on-line capillary electrophoresis/nanoelectrospray ionization-quadrupole time of flight-mass spectrometry (CE/nanoESI-QTOF-MS) and tandem MS of extended chondroitin sulfate/dermatan (CS/DS) oligosaccharide chains is described. The methodology required the construction of a new sheathless CE/nanoESI-QTOF-MS configuration, its implementation and optimization for the high sensitivity analysis of CS/DS oligosaccharide mixtures from conditioned culture medium of decorin transfected human embryonic kidney (HEK) 293 cells. Under newly established sheathless on-line CE/(-)nanoESI conditions for glycosaminoglycan (GAG) ionization and MS detection, single CS/DS oligosaccharide components of extended chain length and increased sulfation degree were identified. Molecular ions corresponding to species carrying 5 and 6 negative charges could be generated for large GAG oligosaccharide species in the negative ion nanoESI-MS. The optimized on-line conditions enabled the detection of molecular ions assigned to oversulfated tetradeca-, octadeca-, and eicosasaccharide CS/DS molecules, which represent the category of largest sulfated GAG-derived oligosaccharides evidenced by CE/ESI-MS. By on-line CE/ESI tandem MS in data-dependent acquisition mode the oversulfated eicosasaccharide species could be sequenced and the localization of the additional sulfate group along the chain could be determined.
authors.affiliation: Institute for Medical Physics and Biophysics, Biomedical Analysis Department.
authors.forename: Alina
authors.initials: A
authors.lastname: Zamfir
authors.affiliation: Institute of Physiological Chemistry and Pathobiochemistry, UKM, University of Münster, Münster, Germany.
authors.forename: Daniela G
authors.initials: DG
authors.lastname: Seidler
authors.affiliation: Institute of Physiological Chemistry and Pathobiochemistry, UKM, University of Münster, Münster, Germany.|Matrix Biology and Tissue Repair Research Unit, University of Wales, Medical College, Dental School, Cardiff, UK.
authors.forename: Elke
authors.initials: E
authors.lastname: Schönherr
authors.affiliation: Institute of Physiological Chemistry and Pathobiochemistry, UKM, University of Münster, Münster, Germany.
authors.forename: Hans
authors.initials: H
authors.lastname: Kresse
authors.affiliation: Institute for Medical Physics and Biophysics, Biomedical Analysis Department.
authors.forename: Jasna
authors.initials: J
authors.lastname: Peter-Katalinic
mesh_terms: D002460:Cell Line; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D006025:Glycosaminoglycans / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D013431:Sulfates / Q000032:analysis / Q000737:chemistry
chemical_list: D006025:Glycosaminoglycans; D009844:Oligosaccharides; D013431:Sulfates