Real-time, label-free characterization of oligosaccharide-binding proteins using carbohydrate microarrays and an ellipsometry-based biosensor.

Carbohydrates present on cell surfaces mediate cell behavior through interactions with other biomolecules. Due to their structural complexity, diversity, and heterogeneity, it is difficult to fully characterize a variety of carbohydrates and their binding partners. As a result, novel technologies for glycomics applications have been developed, including carbohydrate microarrays and label-free detection methods. In this paper, we report using the combination of oligosaccharide microarrays and the label-free oblique-incidence reflectivity difference microscopy for real-time characterization of oligosaccharide-binding proteins. Aminated human milk oligosaccharides were immobilized on epoxy-coated glass substrates as microarrays for reactions with Family 1 of solute-binding proteins fromBifidobacterium longumsubsp.infantis(B. infantis). Binding affinities of these protein–oligosaccharide interactions showed preferences of Family 1 of solute-binding proteins to host glycans, which helps in characterizing the complex process of human milk oligosaccharides foraging byB. infantis. [ABSTRACT FROM PUBLISHER]