earticle

영어

Carbohydrate-protein recognition events play key roles in physiological and pathological processes. The understanding of these biomolecular interactions provides deep insight into glycan-mediated biological processes and enables the development of more effective therapeutic agents and diagnostic tools. Carbohydrate microarrays, which are composed of diverse glycans orderly and densely attached to a single chip substrate, have been extensively developed as reliable and efficient tools for the rapid analysis of carbohydrate-based biomolecular interactions. The most general method for construction of these microarrays involves site-specific and covalent immobilization of chemically modified glycans to properly derivatized surfaces. This immobilization strategy requires the use of properly functionalized sugars, which are typically prepared by multi-step synthetic sequences. In order to avoid the need for functionalized glycans, immobilization strategies that employ unmodified sugars have been developed. In recent investigations directed at this goal, we have developed a novel, direct, site-specific technique for immobilization of unmodified carbohydrates, including simple carbohydrates, oligosaccharides and polysaccharides, on hydrazide-derivatized surfaces. This method eliminates the need for laborious and tedious glycan derivatization. To demonstrate the scope and applicability of this approach, carbohydrate microarrays containing a variety of glycans (fifty eight glycans) have been constructed by using one-step, direct attachment of free carbohydrates to a hydrazide-coated surface. These microarrays have been employed for analysis of sugar binding specificities of lectins, antibodies and bacterial cells. In addition, this microarray format has been applied to the determination of binding affinities between proteins and glycans.