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We describe a simple and rapid chemical strategy for preparing carbohydrate microarrays and utilize these arrays for the characterization of carbohydrate-protein interactions. Carbohydrate arrays were prepared by reductive amination-mediated immobilization of carbohydrate-aminophenyl ethylamine conjugates to aldehyde-activated glass slide. We showed lectins bound specifically to immobilized carbohydrates not only in arrays of homologous disaccharide but also in arrays of heterogeneous disaccharide by using fluorescence imaging system. Carbohydrate microarrays presenting nine disaccharides were then evaluated by profiling the
binding specificities of several lectins. We propose that this array can facilitate analysis of carbohydrate–protein interactions and identification of novel carbohydrate-binding proteins, characterization of carbohydrate processing enzymes, profiling of the binding specificity of antibodies, studies on carbohydrate-mediated cell recognition events and detection of pathogens for diagnosis, deciphering the sugar code (structures of glycans), and discovery of novel inhibitors of protein–carbohydrate interactions and carbohydrate-processing enzymes.