Oinam Lalhaba, Tateno Hiroaki
Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
Methods Mol Biol. 2022;2460:25-32. doi: 10.1007/978-1-0716-2148-6_2.
Glycan microarray is an essential tool to study glycan-binding proteins called lectins. Using glycan microarrays, glycan-binding specificity can be analyzed by incubation with an array in which a series of glycans are immobilized. Various research groups in the world have developed glycan microarray. Among them, our glycan microarray has two unique points: one is the incorporation of the evanescent-field fluorescence detection system, and another is the use of multivalent glycopolymers. These two unique properties allow high-sensitive detection from a relatively limited amount of only nanograms of lectins, which could even be applied in crude samples such as cell lysates and cell culture media. Thus, this system is suitable for the first screening of lectins, lectin-like molecules, lectin candidates, and lectin mutants. Here we describe the protocols to analyze glycan-binding specificity of lectins using our glycan microarray system.
聚糖微阵列是研究称为凝集素的聚糖结合蛋白的重要工具。使用聚糖微阵列，可以通过与固定有一系列聚糖的阵列孵育来分析聚糖结合特异性。世界上各个研究小组都开发了聚糖微阵列。其中，我们的聚糖微阵列有两个独特之处：一是采用了倏逝场荧光检测系统，另一个是使用了多价糖聚合物。这两个独特的特性使得仅从相对少量的纳克级凝集素中就能进行高灵敏度检测，甚至可以应用于细胞裂解物和细胞培养基等粗样品。因此，该系统适用于凝集素、凝集素样分子、凝集素候选物和凝集素突变体的初次筛选。在此，我们描述了使用我们的聚糖微阵列系统分析凝集素聚糖结合特异性的实验方案。
