Construction of Carbohydrate Microarrays by Using One-Step, Direct Immobilizations of Diverse Unmodified Glycans on Solid Surfaces
Construction of Carbohydrate Microarrays by Using One-Step, Direct Immobilizations of Diverse Unmodified Glycans on Solid Surfaces
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DOI：
10.1021/bc800442z
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发表时间：
2009-01-01
影响因子：
4.7
通讯作者：
Shin, Injae
中科院分区：
文献类型：
--
作者：
Park, Sungjin;Lee, Myung-Ryul;Shin, Injae
糖微阵列作为高通量分析工具在研究糖介导的生物过程中受到了极大的关注。大多数用于制造聚糖微阵列的方法依赖于将修饰的聚糖固定在适当衍生化的表面上。这种固定化策略需要修饰聚糖的可用性，其合成在许多情况下是耗时且困难的。我们已经开发了一种简单而直接的固定化技术，涉及一个步骤，不同的未修饰的聚糖的酰肼衍生的玻璃表面的位点特异性连接。为了证明这种直接固定方法的一般性，我们研究了其用于构建含有各种聚糖的碳水化合物微阵列。蛋白质和细胞结合实验的结果表明，通过使用这种方法制备的聚糖微阵列，适用于快速评价聚糖介导的生物分子相互作用和碳水化合物和蛋白质之间的定量结合亲和力的测定。
Carbohydrate microarrays have received great attention as high-throughput analytic tools in studies of carbohydrate-mediated biological processes. Most of the methods employed to fabricate glycan microarrays rely on the immobilization of modified glycans on the properly derivatized surfaces. This immobilization strategy requires the availability of modified glycans whose syntheses in many cases are time-consuming and difficult. We have developed a simple and direct immobilization technique that involves a one-step, site-specific attachment of diverse unmodified glycans to the hydrazide-derivatized glass surface. To demonstrate the generality of this direct immobilization method, we examined its use for the construction of carbohydrate microarrays containing a variety of glycans. The results of protein and cell-binding experiments indicate that the glycan microarrays, prepared by using this methodology, are applicable to the rapid evaluation of glycan-mediated biomolecular interactions and the determination of quantitative binding affinities between carbohydrates and proteins.