National Institute of Advanced Science and Technology, Central-2, 1-1-1, Umezono, Tsukuba, Ibaraki 305-8568, Japan.
Chem Soc Rev. 2013 May 21;42(10):4443-58. doi: 10.1039/c3cs35419a. Epub 2013 Feb 26.
The lectin microarray is a novel platform for glycan analysis, having emerged only in recent years. Unlike other conventional methods, e.g., liquid chromatography and mass spectrometry, it enables rapid and high-sensitivity profiling of complex glycan features without the need for liberation of glycans. Target samples include an extensive range of glycoconjugates involved in cells, tissues, body fluids, as well as synthetic glycans and their mimics. Various procedures for rapid differential glycan profiling have been developed for glycan-related biomarkers. Such glycoproteomics targeting allows precise diagnosis of chronic diseases potentially related to cancer. Application of this method to evaluation of various types of stem cells resulted in the discovery of a new pluripotent cell-specific glycan marker. To explore this technology a more fundamental and extensive understanding of lectins is necessary in relation to the structural uniqueness of glycans. In this chapter, the essence of the lectin microarray is described with some focus on an evanescent-field-activated fluorescence detection principle as a system to achieve in situ (i.e., washing free) aqueous-phase observation under equilibrium conditions. The developed lectin microarray system allows even researchers with poor experience in glycan profiling to perform extensive high-throughput analysis targeting various forms of glycans and even cells.
凝集素微阵列是一种新型聚糖分析平台,近年来才崭露头角。与其他传统方法(例如液相色谱和质谱)不同,它无需释放聚糖,即可实现对复杂聚糖特征的快速和高灵敏度分析。目标样本包括广泛涉及细胞、组织、体液的糖缀合物,以及合成聚糖及其类似物。为了寻找与聚糖相关的生物标志物,已经开发出了各种用于快速差异糖分析的方法。这种针对糖蛋白组学的方法可实现与癌症相关的慢性疾病的精确诊断。将该方法应用于各种类型的干细胞评估,发现了一种新的多能细胞特异性聚糖标记物。为了探索这项技术,有必要更深入地了解与聚糖结构独特性相关的凝集素。本章介绍了凝集素微阵列的本质,并重点介绍了作为在平衡条件下实现无洗涤原位(即在溶液中)观察的一种系统的瞬逝场激活荧光检测原理。开发的凝集素微阵列系统甚至允许经验不足的聚糖分析人员针对各种形式的聚糖甚至细胞进行广泛的高通量分析。