A chip‐based amide‐HILIC LC/MS platform for glycosaminoglycan glycomics profiling

A key challenge to investigations into the functional roles of glycosaminoglycans (GAGs) in biological systems is the difficulty in achieving sensitive, stable, and reproducible mass spectrometric analysis. GAGs are linear carbohydrates with domains that vary in the extent of sulfation, acetylation, and uronic acid epimerization. It is of particular importance to determine spatial and temporal variations of GAG domain structures in biological tissues. In order to analyze GAGs from tissue, it is useful to couple MS with an on‐line separation system. The purposes of the separation system are both to remove components that inhibit GAG ionization and to enable the analysis of very complex mixtures. This contribution presents amide–silica hydrophilic interaction chromatography (HILIC) in a chip‐based format for LC/MS of heparin, heparan sulfate (HS) GAGs. The chip interface yields robust performance in the negative ion mode that is essential for GAGs and other acidic glycan classes while the built‐in trapping cartridge reduces background from the biological tissue matrix. The HILIC chromatographic separation is based on a combination of the glycan chain lengths and the numbers of hydrophobic acetate (Ac) groups and acidic sulfate groups. In summary, chip based amide‐HILIC LC/MS is an enabling technology for GAG glycomics profiling.

[1]  I. Kakizaki,et al.  Enzymatic reconstruction of dermatan sulfate. , 2000, Biochemical and biophysical research communications.

[2]  H. Yin,et al.  Microfluidic chip for peptide analysis with an integrated HPLC column, sample enrichment column, and nanoelectrospray tip. , 2005, Analytical chemistry.

[3]  N. Packer,et al.  Graphitized carbon LC-MS characterization of the chondroitin sulfate oligosaccharides of aggrecan. , 2007, Analytical chemistry.

[4]  C. Dowd,et al.  Heparan Sulfate Mediates bFGF Transport through Basement Membrane by Diffusion with Rapid Reversible Binding* , 1999, The Journal of Biological Chemistry.

[5]  S. Selleck,et al.  Structural analysis of glycosaminoglycans in animals bearing mutations in sugarless, sulfateless, and tout-velu. Drosophila homologues of vertebrate genes encoding glycosaminoglycan biosynthetic enzymes. , 2000, The Journal of biological chemistry.

[6]  B. Strauss,et al.  The role of perlecan in arterial injury and angiogenesis. , 2004, Cardiovascular research.

[7]  R. Langer,et al.  Enzymatic degradation of glycosaminoglycans. , 1995, Critical reviews in biochemistry and molecular biology.

[8]  J. Leary,et al.  Compositional analysis and quantification of heparin and heparan sulfate by electrospray ionization ion trap mass spectrometry. , 2003, Analytical chemistry.

[9]  Jeffrey D. Esko,et al.  Heparan sulphate proteoglycans fine-tune mammalian physiology , 2007, Nature.

[10]  Niclas G Karlsson,et al.  Negative ion graphitised carbon nano-liquid chromatography/mass spectrometry increases sensitivity for glycoprotein oligosaccharide analysis. , 2004, Rapid communications in mass spectrometry : RCM.

[11]  J. Hirsh,et al.  Heparin Binding Proteins Contribution to Heparin Rebound After Cardiopulmonary Bypass , 1993, Circulation.

[12]  A. Ori,et al.  The heparanome and regulation of cell function: structures, functions and challenges. , 2008, Frontiers in bioscience : a journal and virtual library.

[13]  J. Esko,et al.  The sweet and sour of cancer: glycans as novel therapeutic targets , 2005, Nature Reviews Cancer.

[14]  J. Cipollo,et al.  A glycomics platform for the analysis of permethylated oligosaccharide alditols , 2007, Journal of the American Society for Mass Spectrometry.

[15]  S. Selleck,et al.  Order out of chaos: assembly of ligand binding sites in heparan sulfate. , 2002, Annual review of biochemistry.

[16]  Oliver Hobert,et al.  The molecular diversity of glycosaminoglycans shapes animal development. , 2006, Annual review of cell and developmental biology.

[17]  Joseph Zaia,et al.  Size-exclusion chromatography of heparin oligosaccharides at high and low pressure. , 2006, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[18]  L. Kjellén,et al.  Regulated Diversity of Heparan Sulfate* , 1998, The Journal of Biological Chemistry.

[19]  Catherine E Costello,et al.  Glycoform quantification of chondroitin/dermatan sulfate using a liquid chromatography-tandem mass spectrometry platform. , 2006, Biochemistry.

[20]  R. Dwek,et al.  1-N-glycyl beta-oligosaccharide derivatives as stable intermediates for the formation of glycoconjugate probes. , 1992, Biochemistry.

[21]  T. Hayakawa,et al.  Application of liquid chromatography/mass spectrometry and liquid chromatography with tandem mass spectrometry to the analysis of the site-specific carbohydrate heterogeneity in erythropoietin. , 2000, Analytical biochemistry.

[22]  N. Karamanos,et al.  Derivatization of carbohydrates for chromatographic, electrophoretic and mass spectrometric structure analysis. , 2003, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[23]  Anders Hjerpe,et al.  Determination of twelve heparin‐ and heparan sulfate‐derived disaccharides as 2‐aminoacridone derivatives by capillary zone electrophoresis using ultraviolet and laser‐induced fluorescence detection , 2002, Electrophoresis.

[24]  I. Kakizaki,et al.  Domain Structure of Chondroitin Sulfate E Octasaccharides Binding to Type V Collagen* , 2002, The Journal of Biological Chemistry.

[25]  N. Packer,et al.  Use of graphitised carbon negative ion LC-MS to analyse enzymatically digested glycosaminoglycans. , 2005, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[26]  I. Kakizaki,et al.  Chimeric glycosaminoglycan oligosaccharides synthesized by enzymatic reconstruction and their use in substrate specificity determination of Streptococcus hyaluronidase. , 2000, Journal of biochemistry.

[27]  Zachary Shriver,et al.  Roles of heparan-sulphate glycosaminoglycans in cancer , 2002, Nature Reviews Cancer.

[28]  N. Leymarie,et al.  Characterization of heparin oligosaccharides binding specifically to antithrombin III using mass spectrometry. , 2008, Biochemistry.

[29]  Balagurunathan Kuberan,et al.  Analysis of heparan sulfate oligosaccharides with ion pair-reverse phase capillary high performance liquid chromatography-microelectrospray ionization time-of-flight mass spectrometry. , 2002, Journal of the American Chemical Society.

[30]  U. Lindahl,et al.  Domain Structure of Heparan Sulfates from Bovine Organs* , 1996, The Journal of Biological Chemistry.

[31]  R. Midura,et al.  Fluorophore-assisted carbohydrate electrophoresis (FACE) of glycosaminoglycans. , 2001, Osteoarthritis and cartilage.

[32]  M. Nugent,et al.  Fibroblast growth factor-2. , 2000, The international journal of biochemistry & cell biology.

[33]  J. Silbert,et al.  Biosynthesis of Chondroitin/Dermatan Sulfate , 2002, IUBMB life.

[34]  Carlito Lebrilla,et al.  Nanoliquid chromatography‐mass spectrometry of oligosaccharides employing graphitized carbon chromatography on microchip with a high‐accuracy mass analyzer , 2005, Electrophoresis.

[35]  J. Zaia,et al.  Compositional analysis of glycosaminoglycans by electrospray mass spectrometry. , 2001, Analytical chemistry.

[36]  J. Zaia,et al.  Comparative glycomics of connective tissue glycosaminoglycans , 2008, Proteomics.

[37]  R. Midura,et al.  Microanalysis of enzyme digests of hyaluronan and chondroitin/dermatan sulfate by fluorophore-assisted carbohydrate electrophoresis (FACE). , 2000, Glycobiology.

[38]  J. Jorgenson,et al.  In-Depth Characterization of Slurry Packed Capillary Columns with 1.0-μm Nonporous Particles Using Reversed-Phase Isocratic Ultrahigh-Pressure Liquid Chromatography , 2004 .

[39]  R. Linhardt,et al.  Heparin-protein interactions. , 2002, Angewandte Chemie.

[40]  J. Henriksen,et al.  On-line size-exclusion chromatography/mass spectrometry of low molecular mass heparin. , 2004, Journal of mass spectrometry : JMS.

[41]  Hongfeng Yin,et al.  The fundamental aspects and applications of Agilent HPLC-Chip. , 2007, Journal of separation science.

[42]  L. M. Likhosherstov,et al.  A new simple synthesis of amino sugar β-d-glycosylamines , 1986 .

[43]  A. Deelder,et al.  Normal-phase nanoscale liquid chromatography-mass spectrometry of underivatized oligosaccharides at low-femtomole sensitivity. , 2004, Analytical chemistry.

[44]  H. Desaire,et al.  Evidence of block and randomly sequenced chondroitin polysaccharides: sequential enzymatic digestion and quantification using ion trap tandem mass spectrometry. , 2001, Analytical chemistry.

[45]  K. Takagaki,et al.  Enzymatic reconstruction of a hybrid glycosaminoglycan containing 6-sulfated, 4-sulfated, and unsulfated N-acetylgalactosamine. , 1999, Biochemical and biophysical research communications.

[46]  T. Hayakawa,et al.  Analysis of carbohydrate heterogeneity in a glycoprotein using liquid chromatography/mass spectrometry and liquid chromatography with tandem mass spectrometry. , 1999, Analytical biochemistry.

[47]  Joseph Zaia,et al.  Tags for the stable isotopic labeling of carbohydrates and quantitative analysis by mass spectrometry. , 2007, Analytical chemistry.

[48]  J. Esko,et al.  Molecular diversity of heparan sulfate. , 2001, The Journal of clinical investigation.

[49]  Peter Roepstorff,et al.  Ion-pairing reversed-phased chromatography/mass spectrometry of heparin. , 2006, Carbohydrate research.

[50]  R. Linhardt,et al.  Structural characterization of human liver heparan sulfate. , 2005, Biochimica et biophysica acta.

[51]  M. Nugent,et al.  Potentiation and inhibition of bFGF binding by heparin: a model for regulation of cellular response. , 2000, Biochemistry.

[52]  H. Akiyama,et al.  Determination of human urinary hyaluronic acid, chondroitin sulphate and dermatan sulphate as their unsaturated disaccharides by high-performance liquid chromatography. , 1991, Journal of chromatography.

[53]  Robert J Linhardt,et al.  Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides* , 2004, Journal of Biological Chemistry.