Quantifying Protein-Carbohydrate Interactions Using Liquid Sample Desorption Electrospray Ionization Mass Spectrometry

  • Yuyu Yao
  • Km Shams-Ud-Doha
  • Rambod Daneshfar
  • Elena N. Kitova
  • John S. Klassen
Research Article


The application of liquid sample desorption electrospray ionization mass spectrometry (liquid sample DESI-MS) for quantifying protein–carbohydrate interactions in vitro is described. Association constants for the interactions between lysozyme and β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-D-GlcNAc and β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-D-GlcNAc, and between a single chain antibody and α-D-Galp-(1 → 2)-[α-D-Abep-(1 → 3)]-α-D-Manp-OCH3 and β-D-Glcp-(1 → 2)-[α-D-Abep-(1 → 3)]-α-D-Manp-OCH3 measured using liquid sample DESI-MS were found to be in good agreement with values measured by isothermal titration calorimetry and the direct ESI-MS assay. The reference protein method, which was originally developed to correct ESI mass spectra for the occurrence of nonspecific ligand-protein binding, was shown to reliably correct liquid sample DESI mass spectra for nonspecific binding. The suitability of liquid sample DESI-MS for quantitative binding measurements carried out using solutions containing high concentrations of the nonvolatile biological buffer phosphate buffered saline (PBS) was also explored. Binding of lysozyme to β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-D-GlcNAc in aqueous solutions containing up to 1× PBS was successfully monitored using liquid sample DESI-MS; with ESI-MS the binding measurements were limited to concentrations less than 0.02 X PBS.

Graphical Abstract


Protein Carbohydrate Affinity Liquid sample desorption electrospray ionization mass spectrometry 



The authors acknowledge the Alberta Glycomics Center and the Natural Sciences and Engineering Research Council of Canada for funding, and thank Professor D. Bundle (University of Alberta) for generously donating oligosaccharides used in this study.

Supplementary material

13361_2014_1008_MOESM1_ESM.docx (4.7 mb)
ESM 1 (DOCX 4825 kb)


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Copyright information

© American Society for Mass Spectrometry 2014

Authors and Affiliations

  • Yuyu Yao
    • 1
  • Km Shams-Ud-Doha
    • 1
  • Rambod Daneshfar
    • 1
  • Elena N. Kitova
    • 1
  • John S. Klassen
    • 1
  1. 1.Alberta Glycomics Center and Department of ChemistryUniversity of AlbertaEdmontonCanada

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