title: Fast screening of glycosaminoglycan disaccharides by fluorophore-assisted carbohydrate electrophoresis (FACE): applications to biologic samples and pharmaceutical formulations.
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.
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, 21100, Varese, Italy.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Karousou
authors.forename: Athanasia P
authors.initials: AP
authors.lastname: Asimakopoulou
authors.forename: Vassiliki
authors.initials: V
authors.lastname: Zafeiropoulou
authors.forename: Manuela
authors.initials: M
authors.lastname: Viola
authors.forename: Luca
authors.initials: L
authors.lastname: Monti
authors.forename: Antonio
authors.initials: A
authors.lastname: Rossi
authors.forename: Alberto
authors.initials: A
authors.lastname: Passi
authors.forename: Nikos
authors.initials: N
authors.lastname: Karamanos
mesh_terms: D000609:Aminoacridines / Q000737:chemistry; D000818:Animals; D002021:Buffers; D002626:Chemistry, Pharmaceutical; D002809:Chondroitin Sulfates / Q000097:blood / Q000652:urine; D002851:Chromatography, High Pressure Liquid; D004187:Disaccharides / Q000032:analysis / Q000097:blood* / Q000652:urine*; D004586:Electrophoresis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000032:analysis / Q000097:blood* / Q000302:isolation & purification / Q000652:urine*; D006493:Heparin / Q000302:isolation & purification; D006497:Heparan Sulfate / Q000302:isolation & purification; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis; D007903:Lens Capsule, Crystalline / Q000378:metabolism; D051379:Mice; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D051381:Rats
chemical_list: D000609:Aminoacridines; D002021:Buffers; D002809:Chondroitin Sulfates; D004187:Disaccharides; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D004364:Pharmaceutical Preparations; C072515:2-aminoacridone