Development of a lectin microarray based on an evanescent-field fluorescence principle.

Abstract

To investigate protein-carbohydrate interactions in a comprehensive and high-throughput manner, carbohydrate biosensors including microarrays have recently attracted increased attention. In this context, carbohydrate and lectin microarrays are emerging as techniques to meet such requisites. However, most of these methods adopt a conventional immuno-detection system, which requires repetitive washing steps before detection. Since lectin-carbohydrate interactions are relatively weak compared with those between antigens and antibodies, a more precise analytical method, which does not require any washing step, is desirable. We describe here a novel platform for lectin microarray that enables direct observation of lectin-carbohydrate interactions under equilibrium conditions, on the basis of an evanescent-field fluorescence-assisted detection principle. This method allows the analysis of a panel of glycoproteins (glycopeptides) in an extremely sensitive manner. The system also allows real-time observation of lectin-glycoprotein interactions in an aqueous phase. No washing procedures are required, thus relatively weak interactions are detectable. The described lectin microarray is expected to be useful for various fields of glycomics requiring high-throughput analysis of not only purified glycoproteins but also of crude samples.