三维功能糖芯片制备的高效及通用策略

糖芯片具有检测样品用量少、特异性高和高通量等优点，为糖生物学和糖组学的研究提供了一种新的技术平台，对糖-蛋白质相互作用的研究及功能糖组学的发展具有重要的意义。但是，目前糖芯片制备中有三大难点：糖链结构的复杂多样导致糖探针获取困难、糖-蛋白的结合力弱影响检测灵敏度、缺乏制备糖芯片的高效及通用的方法。基于此，本项目计划：1）应用n+1羟基控制转型技术合成多种单糖和二糖、三糖等低分子量寡糖衍生物，为高效合成糖探针提供一个新途径；2）采用点击化学反应控制合成不同的三维芯片树（枝）形聚合物基质，利用"糖簇效应"解决糖-蛋白相互作用弱、检测灵敏度低的问题；3）在三维基质芯片表面引入全氟代苯叠氮基团，通过光引发插入反应直接连接任意种类糖探针，为制备三维功能糖芯片提供一种既高效简便又通用的新方法。

Carbohydrate microarray technology is a novel tool emerging at the frontier of glycomics and glycobiology, having the advantages of small amount of sample needed, high specificity and high throughput, is of great significance to the study of carbohydrate - protein interactions and the development of glycomics. However, there are some problems hindered its preparation and application: difficult access to the various carbohydrates due to the complexity and diversity of their structures; low sensitivity to the weak interactions of carbohydrate - protein; and lack of efficient and universal method for preparation. Based on these, we will, 1)develope a new method for the efficient syntheses of a variety of derivatives of monosaccharides, disaccharides,and trisaccharides in only a few steps by using the n+1 hydroxy reversion technique; 2) in order to improve the sensitivity of carbohydrate microarray, synthesize 3-D matrices by using click chemistry to fabricate dendrimers on the surface of chip for gaining the sugar cluster effect on amplificatin of the weak interactions of carbohydrate-protein; 3) modify the surface of the 3-D matrices with 4-azido-tetrafulorobezoic acid to endow it with the property of covalent connection to any type of carbohydrate probes with no further chemical modification needed. This suggests an efficient and universal fabrication of carbohydrate microarrays.