Fabrication of carbohydrate microarrays on poly(2-hydroxyethyl methacrylate)-cyanuric chloride-modified substrates for analysis of carbohydrate-lectin interactions
Carbohydrate microarrays provide a powerful tool for high-throughput analysis of carbohydrate-mediated interactions. In this work, they were fabricated on poly(2-hydroxyethyl methacrylate) (pHEMA)-cyanuric chloride (CC)-modified substrates for analysis of carbohydrate-lectin interactions. pHEMA polymer provides an anti-fouling surface and CC linker allows the covalent immobilization of intact carbohydrates. Carbohydrates of different sizes (mannose, glucose, galactose, maltose, and dextran) were successfully immobilized on the proposed substrates and their recognition specificity with concanavalin A (ConA) were highly conserved. Compared with a 4-hydroxyphenylacetic acid (HPA)-CC-based microarray, the proposed pHEMA-CC-based microarray exhibits better anti-nonspecific adsorption ability, higher carbohydrate loading capability, stronger multivalent recognition and lower limit of detection of both spotted carbohydrates and lectin in solution without any further signal amplification. The application of the proposed microarray was validated by studying carbohydrate-influenza A virus H1N1 hemagglutinin interactions and fabricating microarrays containing carbohydrates of different properties. It is extendible to study interactions of more carbohydrates and different lectins, and will meet wider needs in bioanalytical and biomedical application.