Evaluation of glycan-binding specificity by glycoconjugate microarray with an evanescent-field fluorescence detection system.

The glycan microarray is now an essential tool used to study lectins. With this technique, glycan-binding specificity can be easily assessed by incubation with an array immobilizing a series of glycans. Glycan microarrays have been developed by numerous research groups around the world. Among the available microarrays, our glycan microarray has two unique characteristics: one is the incorporation of an evanescent-field fluorescence detection system and the other is the use of multivalent glycopolymers. These two unique properties allow the highly sensitive detection of only nanogram quantities of lectins even in crude samples such as cell lysates and cell culture media. Thus, this system is suitable for the initial screening of lectins, lectin-like molecules, lectin candidates, and lectin mutants. Here I describe the protocols employed to analyze the glycan-binding specificity of lectins using our glycan microarray system.