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Improving N-glycan Coverage Using HPLC-MS with Electrospray Ionization at Subambient Pressure

Overview
Journal Anal Chem
Specialty Chemistry
Date 2012 Oct 3
PMID 23025344
Citations 7
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Abstract

Human serum glycan profiling with mass spectrometry (MS) has been employed to study several disease conditions and is demonstrating promise in, for example, clinical biomarker discovery. However, the low glycan ionization efficiency and the large dynamic range of glycan concentrations in human sera can hinder comprehensive profiling. In particular, large glycans are problematic because they are present at low concentrations and are prone to fragmentation. Here we show that, following liquid chromatographic separation on graphite columns, subambient pressure ionization with nanoelectrospray (SPIN)-MS can expand the serum glycome profile in comparison with the conventional atmospheric pressure electrospray ionization (ESI)-MS with a heated capillary inlet. Notably, the ions generated by the SPIN interface were observed at higher charge states for approximately half of the annotated glycans. Out of a total of 130 detected glycans, 34 were only detected with the SPIN-MS, resulting in improved coverage of glycan families as well as of glycans with larger numbers of labile monosaccharides.

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References
1.
Kronewitter S, De Leoz M, Strum J, An H, Dimapasoc L, Guerrero A . The glycolyzer: automated glycan annotation software for high performance mass spectrometry and its application to ovarian cancer glycan biomarker discovery. Proteomics. 2012; 12(15-16):2523-38. PMC: 3567444. DOI: 10.1002/pmic.201100273. View

2.
Iavarone A, Jurchen J, Williams E . Supercharged protein and peptide ions formed by electrospray ionization. Anal Chem. 2001; 73(7):1455-60. PMC: 1414801. DOI: 10.1021/ac001251t. View

3.
Hua S, An H, Ozcan S, Ro G, Soares S, deVere-White R . Comprehensive native glycan profiling with isomer separation and quantitation for the discovery of cancer biomarkers. Analyst. 2011; 136(18):3663-71. PMC: 3331797. DOI: 10.1039/c1an15093f. View

4.
Kita Y, Miura Y, Furukawa J, Nakano M, Shinohara Y, Ohno M . Quantitative glycomics of human whole serum glycoproteins based on the standardized protocol for liberating N-glycans. Mol Cell Proteomics. 2007; 6(8):1437-45. DOI: 10.1074/mcp.T600063-MCP200. View

5.
Sterling H, Kintzer A, Feld G, Cassou C, Krantz B, Williams E . Supercharging protein complexes from aqueous solution disrupts their native conformations. J Am Soc Mass Spectrom. 2011; 23(2):191-200. PMC: 3265691. DOI: 10.1007/s13361-011-0301-y. View