title: An automated mass spectrometry-based screening method for analysis of sulfated glycosaminoglycans.
abstract: Glycosaminoglycans (GAGs) are linear polysaccharides, consisting of repeated disaccharide units, attached to core proteins in all multicellular organisms. Chondroitin sulfate (CS) and dermatan sulfate (DS) constitute a subgroup of sulfated GAGs for which the degree of sulfation varies between species and tissues. One major goal in GAG characterization is to correlate structure to function. A common approach is to exhaustively degrade the GAG chains and thereafter determine the amount of component disaccharide units. In large-scale studies, there is a need for high-throughput screening methods since existing methods are either very time- or samples consuming. Here, we present a new strategy applying MALDI-TOF MS in positive ion mode for semi-qualitative and quantitative analysis of CS/DS derived disaccharide units. Only a few picomoles of sample are required per analysis and 10 samples can be analyzed in 25 min, which makes this approach an attractive alternative to many established assay methods. The total CS/DS concentration in 19 samples derived from Caenorhabditis elegans and mammalian tissues and cells was determined. The obtained results were well in accordance with concentrations determined by a standard liquid chromatography-based method, demonstrating the applicability of the method for samples from various biological matrices containing CS/DS of different sulfation degrees.
authors.affiliation: Department of Chemistry-BMC, Uppsala University, Box 599, 751 24 Uppsala, Sweden; Science for Life Laboratory, Uppsala University, Box 599, 751 24 Uppsala, Sweden.
authors.forename: Nadezda
authors.initials: N
authors.lastname: Kiselova
authors.affiliation: Department of Medical Biochemistry and Microbiology, Uppsala University, Box 582, 751 23 Uppsala, Sweden.
authors.forename: Tabea
authors.initials: T
authors.lastname: Dierker
authors.affiliation: Department of Medical Biochemistry and Microbiology, Uppsala University, Box 582, 751 23 Uppsala, Sweden.
authors.forename: Dorothe
authors.initials: D
authors.lastname: Spillmann
authors.affiliation: Department of Chemistry-BMC, Uppsala University, Box 599, 751 24 Uppsala, Sweden; Science for Life Laboratory, Uppsala University, Box 599, 751 24 Uppsala, Sweden. Electronic address: margareta.ramstrom@kemi.uu.se.
authors.forename: Margareta
authors.initials: M
authors.lastname: Ramström
mesh_terms: D000818:Animals; D006025:Glycosaminoglycans / Q000032:analysis* / Q000737:chemistry*; D012371:Robotics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013431:Sulfates
chemical_list: D006025:Glycosaminoglycans; D013431:Sulfates; C013786:A73025
keywords: Bioanalytical methods; Chondroitin sulfate; Dermatan sulfate; Glycosaminoglycan; Mass spectrometry; Matrix-assisted laser desorption/ionization-time of flight mass spectrometry