Tseng Susan Y, Wang Cheng-Chi, Lin Chin-Wei, Chen Cheng-Lung, Yu Wen-Yueh, Chen Chung-Hsuan, Wu Chung-Yi, Wong Chi-Huey
Genomics Research Center, Academia Sinica, Taiwan.
Chem Asian J. 2008 Sep 1;3(8-9):1395-405. doi: 10.1002/asia.200800154.
We have developed a novel method of immobilizing glycans onto aluminum-coated glass (ACG) slides for potential use in disease diagnosis and drug discovery. The quality of these sugar chips can be assessed by mass spectrometry and fluorescence measurements with high sensitivity. The unique properties of ACG slides include: 1) the metal oxide layer on the surface can be activated for grafting organic compounds such as modified oligosaccharides; 2) the surface remains electrically conductive, and the grafted oligosaccharides can be simultaneously characterized by mass spectrometry and carbohydrate-binding assay; and 3) the slides are more sensitive than transparent glass slides in binding analysis. To demonstrate this, we synthesized a model compound of mannose with a built-in photocleavable linker bound to the ACG slide surface. The molecular weight of the grafted mannose was identified by mass spectrometry, and the slide was subjected to biotinylated ConA binding followed by Cy3-tagged streptavidin detection. This method was further extended to the preparation of glycan arrays containing lactose and the cancer antigen Globo H.
我们开发了一种将聚糖固定在镀铝玻璃(ACG)载玻片上的新方法,有望用于疾病诊断和药物发现。这些糖芯片的质量可以通过质谱和荧光测量进行高灵敏度评估。ACG载玻片的独特性质包括:1)表面的金属氧化物层可被活化以接枝有机化合物,如修饰的寡糖;2)表面保持导电,接枝的寡糖可同时通过质谱和碳水化合物结合测定进行表征;3)在结合分析中,这些载玻片比透明玻璃载玻片更灵敏。为了证明这一点,我们合成了一种带有可光裂解连接子的甘露糖模型化合物,并将其结合到ACG载玻片表面。通过质谱鉴定接枝甘露糖的分子量,然后对载玻片进行生物素化伴刀豆球蛋白A结合,接着用Cy3标记的链霉亲和素进行检测。该方法进一步扩展到含有乳糖和癌抗原Globo H的聚糖阵列的制备。