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一种用于分析碳水化合物-蛋白质相互作用的功能性碳水化合物芯片平台。

A functional carbohydrate chip platform for analysis of carbohydrate-protein interaction.

机构信息

Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea.

出版信息

Nanotechnology. 2010 May 28;21(21):215101. doi: 10.1088/0957-4484/21/21/215101. Epub 2010 Apr 30.

Abstract

A carbohydrate chip based on glass or other transparent surfaces has been suggested as a potential tool for high-throughput analysis of carbohydrate-protein interactions. Here we proposed a facile, efficient, and cost-effective method whereby diverse carbohydrate types are modified in a single step and directly immobilized onto a glass surface, with retention of functional orientation. We modified various types of carbohydrates by reductive amination, in which reducing sugar groups were coupled with 4-(2-aminoethyl)aniline, which has di-amine groups at both ends. The modified carbohydrates were covalently attached to an amino-reactive NHS-activated glass surface by formation of stable amide bonds. This proposed method was applied for efficient construction of a carbohydrate microarray to analyze carbohydrate-protein interactions. The carbohydrate chip prepared using our method can be successfully used in diverse biomimetic studies of carbohydrates, including carbohydrate-biomolecule interactions, and carbohydrate sensor chip or microarray development for diagnosis and screening.

摘要

基于玻璃或其他透明表面的碳水化合物芯片已被提议作为高通量分析碳水化合物-蛋白质相互作用的潜在工具。在这里,我们提出了一种简便、高效和具有成本效益的方法,通过该方法可以在一步中对各种碳水化合物类型进行修饰,并直接固定在玻璃表面上,同时保留功能取向。我们通过还原胺化修饰了各种类型的碳水化合物,其中还原糖基团与两端都有二胺基团的 4-(2-氨乙基)苯胺偶联。修饰后的碳水化合物通过形成稳定的酰胺键共价连接到氨基反应性 NHS 活化玻璃表面上。该方法已成功应用于高效构建碳水化合物微阵列以分析碳水化合物-蛋白质相互作用。使用我们的方法制备的碳水化合物芯片可成功用于碳水化合物的各种仿生研究,包括碳水化合物-生物分子相互作用,以及用于诊断和筛选的碳水化合物传感器芯片或微阵列的开发。

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