title: Quantitative analysis of oligosaccharides derived from sulfated glycosaminoglycans by nanodiamond-based affinity purification and matrix-assisted laser desorption/ionization mass spectrometry.
abstract: Degraded fragments of sulfated glycosaminoglycans (GAGs) are key reporters for profiling the burden of mucopolysaccharidosis (MPS) disease at baseline and during therapy. Here, we present a high-throughput assay, which combines microwave-assisted degradation, solid-phase affinity purification, and matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS), for quantitative analysis of sulfated oligosaccharides in biological samples. First, sulfated oligosaccharides such as chondroitin-4-sulfate (CS) were efficiently isolated from highly diluted solutions or spiked artificial cerebrospinal fluid (aCSF) using polyarginine-coated nanodiamonds (PA-coated NDs) as affinity sorbents. Next, they were degraded to disaccharides through microwave-assisted methanolysis or enzymatic digestion for subsequent MALDI-TOF MS analysis. The reaction times for GAG depolymerization were significantly reduced from a few hours to less than 7 min under the microwave irradiation. Deuterium-labeled internal standards were then mixed with the CS-derived disaccharides for quantitative analysis by MALDI-TOF MS using the N-(1-naphthyl) ethylenediamine dihydrochloride (NEDC) matrix. The new assay is facile, specific (with distinct chlorine-isotope trait markers), sensitive (with a detection limit of ~70 pg), and potentially useful for clinical diagnosis of MPS.
authors.affiliation: Department of Chemistry, National Taiwan Normal University, Taipei 106, Taiwan.
authors.forename: Chih-Chien
authors.initials: CC
authors.lastname: Hsieh
authors.forename: Jiun You
authors.initials: JY
authors.lastname: Guo
authors.forename: Shain-Un
authors.initials: SU
authors.lastname: Hung
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.forename: Zongxiu
authors.initials: Z
authors.lastname: Nie
authors.forename: Huan-Cheng
authors.initials: HC
authors.lastname: Chang
authors.forename: Chih-Che
authors.initials: CC
authors.lastname: Wu
mesh_terms: D006025:Glycosaminoglycans / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry*; D009844:Oligosaccharides / Q000032:analysis*; D010455:Peptides / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013431:Sulfates / Q000737:chemistry*
chemical_list: D006025:Glycosaminoglycans; D009844:Oligosaccharides; D010455:Peptides; D013431:Sulfates; C015462:polyarginine