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Fast Screening of Glycosaminoglycan Disaccharides by Fluorophore-Assisted Carbohydrate Electrophoresis (FACE): Applications to Biologic Samples and Pharmaceutical Formulations

  • Evgenia Karousou
  • Athanasia P. Asimakopoulou
  • Vassiliki Zafeiropoulou
  • Manuela Viola
  • Luca Monti
  • Antonio Rossi
  • Alberto Passi
  • Nikos KaramanosEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1229)

Abstract

Hyaluronan (HA), chondroitin sulfate (CS), and heparan sulfate (HS) are glycosaminoglycans (GAGs) with a great importance in biological processes as they participate in functional cell properties, such as migration, adhesion, and proliferation. A perturbation of the quantity and/or the sulfation of GAGs is often associated with pathological conditions. In this chapter, we present valuable and validated protocols for the analysis of HA-, CS-, and HS-derived disaccharides after derivatization with 2-aminoacridone and by using the fluorophore-assisted carbohydrate electrophoresis (FACE). FACE is a well-known technique and a reliable tool for a fast screening of GAGs, as it is possible to analyze 16 samples at the same time with one electrophoretic apparatus. The protocols for the gel preparation are based on the variations of the acrylamide/bisacrylamide and buffer concentrations. Different approaches for the extraction and purification of the disaccharides of various biologic samples and pharmaceutical preparations are also stressed.

Key words

Glycosaminoglycans Disaccharides Polyacrylamide gel electrophoresis 2-Aminoacridone Enzymic treatment 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Evgenia Karousou
    • 1
  • Athanasia P. Asimakopoulou
    • 2
  • Vassiliki Zafeiropoulou
    • 2
  • Manuela Viola
    • 1
  • Luca Monti
    • 3
  • Antonio Rossi
    • 3
  • Alberto Passi
    • 1
  • Nikos Karamanos
    • 2
    Email author
  1. 1.Department of Surgical and Morphological SciencesUniversity of InsubriaVareseItaly
  2. 2.Laboratory of Biochemistry, Department of ChemistryUniversity of PatrasPatrasGreece
  3. 3.Department of Molecular Medicine, Unit of BiochemistryUniversity of PaviaPaviaItaly

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