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Mass Spectrometry of Glycoproteins pp 145–169Cite as

Separation and Identification of Glycoforms by Capillary Electrophoresis with Electrospray Ionization Mass Spectrometric Detection

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 951))

Abstract

Capillary electrophoresis (CE) is a resourceful and versatile separation method for the analysis of complex carbohydrate mixtures. In combination with electrospray ionization (ESI) mass spectrometry (MS), CE enables fast, sensitive, and efficient separations for the accurate identification of a large variety of glycoform mixture types. In this chapter several reliable off- and on-line CE-based methods for the analysis of glycoforms with ESI MS/MS are presented. The first part of this chapter is dedicated to the application of off-line CE/ESI MS to complex mixtures of O-glycopeptides and mixtures of proteoglycan-derived O-glycans, i.e., glycosaminoglycans such as depolymerized hybrid chains of chondroitin sulfate (CS) and dermatan sulfate (DS). Procedures for off-line fractionation of these heterogeneous mixtures followed by ESI MS screening and sequencing of single glycoforms by collision-induced dissociation (CID) at low energies are also described. Ample sections are further devoted to on-line CE/ESI MS technique and its application to separation and identification of O-glycopeptides and CS/DS oligosaccharides. The concept and construction principles of two different sheathless CE/ESI MS interfaces together with the protocols to be applied for successful on-line analysis of O-glycopeptides and CS/DS oligosaccharides are presented in details in the last part of the chapter.

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Acknowledgments

The authors are grateful to the Romanian National Authority for Scientific Research, CNCS-UEFISCDI (Grants No. PN- II-ID-PCE-2011-3-0047, PN-II-RU-TE-2011-2-0008 and POSDRU/89/1.5/S/60746) for financial support.

Author information

Authors and Affiliations

  1. Department of Chemical and Biological Sciences, “Aurel Vlaicu” University of Arad, Timisoara, Romania

    Alina D. Zamfir & Corina Flangea

  2. Department of Biochemistry, “Victor Babes” University of Medicine and Pharmacy, Timisoara, Romania

    Alina Serb & Eugen Sisu

  3. Neuroscience Laboratory, “Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania

    Ana-Maria Zagrean

  4. Institute of Analytical Chemistry and Food Chemistry, University of Vienna, Vienna, Austria

    Andreas M. Rizzi

  5. Chemistry Institute of the Romanian Academy, Timisoara, Romania

    Eugen Sisu

Authors
  1. Alina D. Zamfir
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  2. Corina Flangea
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  3. Alina Serb
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  4. Ana-Maria Zagrean
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  5. Andreas M. Rizzi
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  6. Eugen Sisu
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Corresponding author

Correspondence to Alina D. Zamfir .

Editor information

Editors and Affiliations

  1. Medical Center, Department of Biochemistry, University of Texas Southwestern, Harry Hines Blvd. 5323, Dallas, 75390-9038, Texas, USA

    Jennifer J. Kohler

  2. Medical Center, Dallas, University of Texas Southwestern, Harry Hines Blvd. 5323, Dallas, 75390-9185, Texas, USA

    Steven M. Patrie

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Zamfir, A.D., Flangea, C., Serb, A., Zagrean, AM., Rizzi, A.M., Sisu, E. (2013). Separation and Identification of Glycoforms by Capillary Electrophoresis with Electrospray Ionization Mass Spectrometric Detection. In: Kohler, J., Patrie, S. (eds) Mass Spectrometry of Glycoproteins. Methods in Molecular Biology, vol 951. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-146-2_11

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  • DOI: https://doi.org/10.1007/978-1-62703-146-2_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-145-5

  • Online ISBN: 978-1-62703-146-2

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