Versatile Glycan Probes for Multiplatform Investigation of Glycan Interactions with Proteins, Viruses, and Cells

Abstract

Abstract Glycan-mediated interactions are vital to development, microbial colonisation, immune signalling, and cancer progression. Glycan microarrays have revolutionised glycobiology by enabling high-throughput analysis of these complex interactions, supported by techniques that reveal kinetics and dynamics in solution or at the cellular level. We introduce multifunctional glycan probes based on a tri-functional Fmoc-Amino-Azido (FAA) linker, enabling multi-platform investigation of glycan-mediated interactions. These FAA probes support glycan presentation on both covalent and non-covalent array platforms, allowing direct comparison of glycan recognition by diverse proteins. Notably, certain viral adhesins and immune lectins show a preference for the non-covalent platform. The azido group allows further functionalisation via ‘click chemistry’, enabling biotinylation for immobilisation on bio-layer interferometry biosensors for influenza virus binding, or fluorescent tagging for flow cytometry analysis of glycan-lectin interactions on cells. These versatile probes offer a unified platform for in-depth interrogation of glycan interactions using complementary approaches, with strong potential to advance glycan-based diagnostics and therapeutics.