Automated Glycan Assembly of (19)F‐labeled Glycan Probes Enables High‐Throughput NMR Studies of Protein–Glycan Interactions

Protein–glycan interactions mediate important biological processes, including pathogen host invasion and cellular communication. Herein, we showcase an expedite approach that integrates automated glycan assembly (AGA) of (19)F‐labeled probes and high‐throughput NMR methods, enabling the study of protein–glycan interactions. Synthetic Lewis type 2 antigens were screened against seven glycan binding proteins (GBPs), including DC‐SIGN and BambL, respectively involved in HIV‐1 and lung infections in immunocompromised patients, confirming the preference for fucosylated glycans (Le(x), H type 2, Le(y)). Previously unknown glycan–lectin weak interactions were detected, and thermodynamic data were obtained. Enzymatic reactions were monitored in real‐time, delivering kinetic parameters. These results demonstrate the utility of AGA combined with (19)F NMR for the discovery and characterization of glycan–protein interactions, opening up new perspectives for (19)F‐labeled complex glycans.