硼酸修飾表面製備醣微陣列晶片的發展及應用

Fabrication of Carbohydrate Microarrays through Boronate Formation

蕭瑄怡(Hsuan-Yi Hsiao)

指導教授 ： 林俊成

國立清華大學/理學院/化學系/碩士(2010年)

摘要

醣微陣列晶片是一種可同時篩選多種醣體、使用較少的樣品，快速且高通量的分析方式。對於醣生物學的研究領域，醣晶片是一種快速且有效分析醣體與生物分子交互作用的方法。在本研究中，根據硼酸能與醣體上的二元醇形成環狀硼酸酯的特性，發展一種新穎且簡便的醣晶片製程，可提供穩定、共價鍵固化的醣晶片。雖然硼酸酯化反應在水溶液中是一種可逆的反應，但由於醣體上許多羥基的多價體效應，以及硼酸酯共價鍵的特性，醣體固化在玻片表面後不容易被洗掉。除了單醣之外，這樣的固化方式適用於各種大小的醣體，且不影響晶片表面上的醣體與蛋白質的結合能力。此外，市售的醣體或是由生物樣品純化的醣體，可直接固化在硼酸修飾的表面，不需要額外的化學修飾。在篩選分析醣體與蛋白質交互作用與定量分析交互作用的親和力強弱的應用中，硼酸修飾的醣晶片提供了一種便利且快速的平台。

關鍵字

微陣列 ； 醣晶片 ； 硼酸

並列摘要

Carbohydrate microarrays, which screen many carbohydrates simultaneously with a minimal consumption of reagents, provide a viable solution for glycobiology research as a reliable and efficient tool for rapid analysis of carbohydrate-biomolecule interactions. We have developed a novel and easy method for fabricating a stable, covalent, and highly active carbohydrate microarray based on boronate formation between the hydroxyl groups of carbohydrate and boronic acid (BA) on the glass surface. Although the formation of boronate is a reversible reaction in aqueous solution, the BA-based carbohydrate microarray resist the stringent wash because of multivalent binding between carbohydrate hydroxyl groups and BAs on the surface. Except monosaccharides, this method is suitable for immobilizing all sizes of carbohydrates with biological binding activity toward the corresponding lectins. In addition, there is no need to perform sugar modification before immobilization. The BA-based carbohydrate microarrays were used to identify carbohydrate-protein interactions and to measure the dissociation constant of corresponding carbohydrate-binding protein.

並列關鍵字

microarray ； carbohydrate ； boronic acid

參考文獻

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