Abstract
We describe a simple and rapid chemical strategy for preparing carbohydrate microarrays and utilize these arrays for the characterization of carbohydrate-protein interactions. Carbohydrate microarrays were prepared by reductive amination-mediated immobilization of amine-modified carbohydrate onto NHS-activated glass slide. The carbohydrate conjugates were confirmed by MALDI-TOF MS and 1H NMR. Carbohydrate microarray presenting six disaccharide were evaluated by profiling the binding specificities of several lectins. The results showed that the array selectively bound the corresponding lectins with relative binding affinities. We identified lectin and carbohydrate detection limit concentration on this array using WGA(Wheat Germ Agglutinin) as lectin and N,N'-diacetylchitobiose(GlcNAcβ-12Man) as carbohydrate. We also evaluated interactions between cholera toxin B subunit and GM1, GM1 analogues such as asialo GM1, GM3 on this array. As a result, GM1 pentasaccharide showed a strong affinity and specificity in interaction, but asialo GM1 and GM3, which is lacking of terminal Neu5Ac and galactose-N-acetylgalactosamine, respectively, showed a low affinity. We proposed that this carbohydrate microarray system could be applicable for feasible studies, like sensing of pathogenic bacterium or detection of toxin protein.
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