AbstractThe importance of multivalency for N‐glycan‐protein interactions has primarily been studied by attachment of minimal epitopes to artificial multivalent scaffold and not in the context of multi‐antennary glycans. N‐glycans can be modified by bisecting GlcNAc, core xylosides and fucosides, and extended N‐acetyl lactosamine moieties. The impact of such modifications on glycan recognition are also not well understood. We describe here a chemoenzymatic methodology that can provide N‐glycans expressed by the parasitic worm S. mansoni having unique epitopes at each antenna and containing core xyloside. NMR, computational and electron microscopy were employed to investigate recognition of the glycans by the human lectin DC‐SIGN. It revealed that core xyloside does not influence terminal epitope recognition. The multi‐antennary glycans bound with higher affinity to DC‐SIGN compared to mono‐valent counterparts, which was attributed to proximity‐induced effective concentration. The multi‐antennary glycans cross‐linked DC‐SIGN into a dense network, which likely is relevant for antigen uptake and intracellular routing.