Fluorophore-assisted carbohydrate electrophoresis (FACE) of glycosaminoglycans

摘要: Abstract Objective Quantitation and analyses of the fine structure glycosaminoglycans are increasingly important for understanding many biological processes, including those most critical skeletal biology. We have developed a novel procedure, fluorophore-assisted carbohydrate electrophoresis (FACE), determination glycosaminoglycan estimation chain length. Design FACE utilizes enzymes that cleave to create products, usually disaccharides, characteristic enzyme specificity. Each cleavage exposes new reducing terminus is fluorotagged by reductive amination with 2-aminoacridone. The tagged products then displayed electrophoresis, identified their migration chemistry, quantitated molar fluorescence. Results class specific each discussed. Specific application technology shown analysis on aggrecan isolated from knee cartilage 5- 68-year-old patients, assessment hyaluronan oligosaccharides. Conclusions powerful tool all four classes obtained wide variety biologic sources. While protocols relative simple, they provide wealth information quantitation in pmole range, structure,