Evaluation of Glycan-Binding Specificity by Glycoconjugate Microarray with an Evanescent-Field Fluorescence Detection...

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.