Fast Screening of Glycosaminoglycan Disaccharides by Fluorophore-Assisted Carbohydrate Electrophoresis (FACE): Applications to Biologic Samples and Pharmaceutical Formulations
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 wordsGlycosaminoglycans Disaccharides Polyacrylamide gel electrophoresis 2-Aminoacridone Enzymic treatment
- 4.Karousou E, Stachtea X, Moretto P, Viola M, Vigetti D, D’Angelo ML, Raio L, Ghezzi F, Pallotti F, De Luca G, Karamanos NK, Passi A (2013) New insights into the pathobiology of Down syndrome—hyaluronan synthase-2 overexpression is regulated by collagen VI alpha2 chain. FEBS J 280:2418–2430PubMedCrossRefGoogle Scholar
- 18.Karousou EG, Viola M, Genasetti A, Vigetti D, Luca GD, Karamanos NK, Passi A (2005) Application of polyacrylamide gel electrophoresis of fluorophore-labeled saccharides for analysis of hyaluronan and chondroitin sulfate in human and animal tissues and cell cultures. Biomed Chromatogr 19:761–765PubMedCrossRefGoogle Scholar