High-performance affinity chromatography of carbohydrate-binding proteins by two-stage separation on a resin carrying a number of oligosaccharides.
Oligosaccharides of ovalbumin were released by hydrazinolysis and converted to the glycamine derivatives by reductive amination. The resultant derivatives were immobilized on an epoxy-activated methacrylate polymer. Application of lectins on the column containing the resultant resin, followed by injection of the competing sugars and detection of the eluate using natural fluorescence, allowed differentiation of micro amounts of the lectins, owing to their high specificity. Stepwise elution with various competing sugars also permitted separation of lectins. Application of this method to serum samples enabled detection of various carbohydrate-binding proteins with specific affinity to the injected sugars. This method, based on two-stage separation at the adsorption and elution stages, was highly specific. It was also rapid, reproducible, and sensitive.
Honda
S
S
Faculty of Pharmaceutical Sciences, Kinki University, Higashi-osaka, Japan.
Suzuki
K
K
Suzuki
S
S
Kakehi
K
K
D000818:Animals; D002241:Carbohydrates; D002352:Carrier Proteins / Q000032:analysis*; D002645:Chickens; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D009844:Oligosaccharides; D012116:Resins, Plant
D002241:Carbohydrates; D002352:Carrier Proteins; D009844:Oligosaccharides; D012116:Resins, Plant