Chemical Microarrays Constructed by Selective Attachment of Hydrazide-Conjugated Substances to Epoxide Surfaces and Thei

Microarray technology has received considerable attention for rapid analysis of biomolecular interactions and high-throughput screening to identify binding partners. An efficient and selective immobilization technique of substances on the surface is essen

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1. Introduction In the postgenomic era, functional studies of proteins and glycans have been extensively performed for biological and biomedical applications. As powerful tools for these studies, microarray-based technologies, such as DNA, carbohydrate, peptide, and protein microarrays, have been developed for accelerating the identification of lead compounds and for genomic, transcriptomic, glycomic, Mahesh Uttamchandani and Shao Q. Yao (eds.), Small Molecule Microarrays: Methods and Protocols, Methods in Molecular Biology, vol. 669, DOI 10.1007/978-1-60761-845-4_16, © Springer Science+Business Media, LLC 2010

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and proteomic research (1–12). In addition, small molecule microarrays have also been exploited as high-throughput methods to identify bioactive compounds that selectively bind to proteins (13–15). Small molecules that modulate biological processes serve as useful tools in the studies of the functions of proteins as well in the development of pharmaceutical agents. These microarray-based technologies facilitate fast, quantitative and simultaneous assessment of a large number of biomolecular interactions using limited quantities of samples. Many biologically relevant compounds possess various functional groups such as alcohols (-OH), amines (-NH2), carboxylic acids (CO2H), and/or thiols (-SH). The major requirement of techniques used to fabricate chemical microarrays is that immobilization of the diversely functionalized compounds to the modified surfaces must be highly selective. As a result, strategies that employ efficient and selective ligation processes would be generally applicable to the preparation of microarrays that contain covalently linked, biologically interesting molecules with diverse functionalities. Several criteria must be met in designing a general method to prepare chemical microarrays. Firstly, the diverse substances possessing specific functional groups that are used for selective reactions with the modified solid surfaces must be easily synthesized by solid-phase synthesis. Secondly, functional groups that selectively react with substances must be easily incorporated on to the solid surfaces. Lastly, the diversely structured and functionalized substances released from a solid support must undergo site-specific covalent immobilization on the modified surfaces. Strategies, which fit these criteria, employ highly chemoselective ligation reactions. The immobilization technique which relies on the use of reactions between hydrazide-containing substances and epoxide-derivatized surfaces is suitable for the fabrication of chemical microarrays (see Fig. 1). The hydrazide groups incorporated into the substances are used as a handle in their solid-phase synthesis. The epoxide-derivatized glass slide is easily created by immersing amine-coated glass slides into a solution of poly(ethylene glycol) diglycidyl ether. Importantly, the reactions between hydrazides and epoxides are chemoselective even in the presence of other functionalities such as thiol and amine grou