ABO Antigens Active Tri- and Disaccharides Microarray to Evaluate C-type Lectin Receptor Binding Preferences

Abstract

Understanding blood group antigen binding preferences for C-type lectin receptors holds promise for modulating immune responses, since several Gram-negative bacteria express blood group antigens as molecular mimicry to evade immune responses. Herein, we report the synthesis of ABO blood group antigen active tri and disaccharides to investigate the binding specificity with various C-type lectin receptors using glycan microarray. The results of binding preferences show that distinct glycosylation on the galactose and fucose motifs are key for C-type lectin receptor binding and that these interactions occur in a Ca2+-dependent fashion.

Practical considerations for printing high-density glycan microarrays to study weak carbohydrate-protein interactions

“…Existing microarray platforms that use NHS-ester chemistry for glycan immobilization print the oligosaccharides on the activated glass slides differently. Different labs successfully use slides from different manufacturers, for example Nexterion H slides from Schott [8,20,21], Codelink slides from Surmodics [8,22], and less commonly also slides from the manufacturer PolyAn [23,24]. Moreover, different printing buffers such as sodium phosphate buffers or phosphate buffers including polyethylenglycol (PEG) 400 [12,17,25,26], different humidity [18,27], and different glycan concentrations are used for printing [19].…”