Covalent immobilization of carbohydrates on sol–gel-coated microplates

Abstract

Carbohydrate microarrays have attracted increasing attention in recent years because of their ability to monitor biologically important protein–carbohydrate interactions in a high-throughput manner. Here we have developed an effective approach to immobilizing intact carbohydrates directly on polystyrene microtiter plates coated with amine-functionalized sol–gel monolayers. Lectin binding was monitored by fluorescence spectroscopy using these covalent arrays of carbohydrates that contained six mono- and di-saccharides on the microplates. In addition, binding affinities of lectin to carbohydrates were also quantitatively analyzed by determining IC₅₀ values of lectin-specific antibody with these arrays. Our results indicate that microplate-based carbohydrate arrays can be efficiently fabricated by covalent immobilization of intact carbohydrates on sol–gel-coated microplates. The microplate-based carbohydrate arrays can be applied for screening of protein–carbohydrate interactions in a high-throughput manner.