Preparation of Carbohydrate Arrays by Using Diels-Alder Reactions with Inverse Electron Demand
Preparation of Carbohydrate Arrays by Using Diels-Alder Reactions with Inverse Electron Demand
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DOI：
10.1002/chem.201200382
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发表时间：
2012-05-01
影响因子：
4.3
通讯作者：
Wittmann, Valentin
中科院分区：
文献类型：
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作者：
Beckmann, Henning S. G.;Niederwieser, Andrea;Wittmann, Valentin
碳水化合物微阵列是一种用于高通量筛选碳水化合物蛋白质相互作用的新兴工具，它代表了许多生物学和医学相关过程的基础。制备碳水化合物阵列的关键步骤是将碳水化合物探针附着到表面。为此，我们将逆电子需求 (DARinv) 的 DielsAlder 反应视为不可逆的化学选择性连接反应。在显示了 DARinv 在溶液中的效率后，我们制备了一系列碳水化合物亲二烯体缀合物，将其印刷到四嗪修饰的载玻片上。荧光标记凝集素的结合实验证明是成功的，并且 DARinv 实现了均匀固定。为了固定非功能化的还原性寡糖，我们开发了一种双功能化学选择性接头，能够通过肟连接将亲双烯体标签附着到寡糖上。获得的缀合物成功固定在载玻片上。所提出的合成碳水化合物衍生物和未受保护的还原寡糖的固定化策略有助于通过化学选择性 DARinv 制备高质量碳水化合物微阵列。这个概念可以很容易地适用于其他生物分子阵列的制备。
Carbohydrate microarrays are an emerging tool for the high-throughput screening of carbohydrateprotein interactions that represent the basis of many biologically and medicinally relevant processes. The crucial step in the preparation of carbohydrate arrays is the attachment of carbohydrate probes to the surface. We examined the DielsAlder reaction with inverse-electron-demand (DARinv) as an irreversible, chemoselective ligation reaction for that purpose. After having shown the efficiency of the DARinv in solution, we prepared a series of carbohydratedienophile conjugates that were printed onto tetrazine-modified glass slides. Binding experiments with fluorescently labeled lectins proved successful and homogeneous immobilization was achieved by the DARinv. For immobilization of nonfunctionalized reducing oligosaccharides we developed a bifunctional chemoselective linker that enabled the attachment of a dienophile tag to the oligosaccharides through oxime ligation. The conjugates obtained were successfully immobilized on glass slides. The presented strategies for the immobilization of both synthetic carbohydrate derivatives and unprotected reducing oligosaccharides facilitate the preparation of high-quality carbohydrate microarrays by means of the chemoselective DARinv. This concept can be readily adapted for the preparation of other biomolecule arrays.