Figure 2.

Human intelectin-1 (hItln-1) binding to monosaccharides. (a) Structures of monosaccharides identified as potential hItln-1 ligands from glycan microarray and evaluated in hItln-1 binding studies. (b) Representative real-time biolayer interferometry sensorgram (BLItz) of competition of hItln-1 binding to immobilized biotinylated Galf with varying concentrations of soluble allyl-α-d-glycero-d-talo-oct-2-ulosonic acid (allyl-α-KO). (c) Competition of soluble monosaccharide epitopes with immobilized β-Galf using BLItz. Endpoint data (710–720 s) from BLItz sensorgrams were averaged and normalized for each competitor concentration and fitted to a one site logIC₅₀ equation (solid lines). IC₅₀ values were determined (Table 1). The allyl glycoside of each compound in the anomer shown in (a) was tested in the competition assay. (d) Evaluation of BSA-conjugated heptose sugars as ligands for hItln-1 using ELISA. Data shown as mean ± SEM (n = 2 technical replicates). Data were fit to a single-site binding equation (solid lines). Kd values could not be determined for l,d-heptose or D, d-heptose. OD, optical density.