Elegant chemistry to directly anchor intact saccharides on solid surfaces used for the fabrication of bioactivity-conserved saccharide microarrays.
Elegant chemistry to directly anchor intact saccharides on solid surfaces used for the fabrication of bioactivity-conserved saccharide microarrays.
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DOI：
10.1021/bc300142s
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发表时间：
2012-05
影响因子：
4.7
通讯作者：
Kai Liang;Y. Chen
中科院分区：
文献类型：
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作者：
Kai Liang;Y. Chen
提出了一种简单的化学策略，并用于建立一种方法，直接锚定完整的生物亲和性良好的固体表面。以三聚氯氰为关键连接剂，通过温度调节的分步反应实现了锚定，并成功地应用于糖芯片的制作。为了证明这一点，将15个不同分子大小的完整的还原性和非还原性酶点在一个三聚氰酸-氯化物修饰的芯片（1.0 × 1.0 cm（2））上，并使其与凝集素反应。正如预期的那样，锚定的甘露糖能够识别它们的靶凝集素，更令人兴奋的是锚定单糖如甘露糖、葡萄糖甚至果糖（与伴刀豆球蛋白A的相互作用很弱）的特异性识别能力的完美保守。这种保守性归因于固定化后保留了锚定结构的原始结构（特别是异头构型），并允许锚定的锚定结构与氰尿酰氯的支架一起旋转和/或在氰尿酰氯的支架上旋转，这使得它们很容易适应锚定优选的空间位置。通过质谱和核磁共振分别表征了三聚氯氰上的预期连接和它们的异头构型的维持。新方法不仅可以突出其保护糖的生物亲和性和普遍适用性，而且还可以突出其优点，易于操作或易于控制的反应和成本效益，由于使用非常便宜的氰尿酰氯。
An easy chemical strategy was proposed and used to establish a method for direct anchoring of intact saccharides on solid surfaces with well conserved bioaffinity. The anchoring was achieved by temperature-modulated stepwise reactions with cyanuric chloride as a key linker, and was successfully applied to the fabrication of saccharide chips. To demonstrate, 15 intact reducing and nonreducing saccharides with various molecular sizes were dotted on a cyanuric-chloride-modified chip (1.0 × 1.0 cm(2)) and made to react with lectins. As expected, the anchored saccharides were capable of recognizing their target lectins, and more exciting were the perfect conservation of the specific recognizing ability of the anchored monosaccharides such as mannose, glucose, and even fructose (interacting only weakly with concanavalin A). This conservation was ascribed to the maintenance of the original structure (especially the anomeric configuration) of saccharides after immobilization and to the allowance of the anchored saccharides to rotate with and/or on the scaffold of cyanuric chloride, which makes them easily adapt to the recognition-preferred spatial position. The expected linkage of saccharides on cyanuric chloride and the maintenance of their anomeric configuration were characterized by mass spectrometry and nuclear magnetic resonance, respectively. The new method can be highlighted not only by its conservation of saccharide bioaffinity and universal applicability but also by its merits of easy manipulation or facile control of the reactions and cost-effectiveness due to the use of extremely cheap cyanuric chloride.