Fig. 2: Directed evolution of omalizumab variants using a disruptive efficiency screen.
a Schematic plot of ID50 vs KD, with two different efficiency trend lines plotted in dotted or dashed lines. Hypothetical changes in the behavior of a disruptive inhibitor “A,” are diagramed to show changes in affinity only (A → B), improvement in affinity and disruptive potency (A → C), or pure improvement in disruptive efficiency (A → D). b Cartoon schematic of standard affinity maturation selection approaches. c Cartoon schematic of a disruptive potency selection pathway. d Cartoon schematic of a disruptive efficiency selection pathway. e Schematic with recombinant constructs used to engineer the IgE:FcεRIα complex. f Structure of displayed anti-IgE agents bound to IgE-Fc3–4 with FcεRIα binding site highlighted. g Yeast-displayed anti-IgE agents stained with a titration of IgE-Fc2–4 (left) or IgE-Fc2–4:FcεRIα (right). h (Left) singlet-cMyc+ anti-IgE yeast stained for disruptive potency selection and anti-cMyc at indicated concentrations. (Right) singlet-cMyc+ yeast stained for disruptive efficiency selection at indicated concentrations. i Schematic of co-binding screen. j Normalized histogram of singlet-cMyc+ yeast anti-IgE controls stained with secondary reagents alone to assess background signal on yeast, binding to bIgE-Fc2–4 alone (100 nM), or co-binding to bIgE-Fc2–4 (100 nM) in complex with FcεRIα-Ova (1 μM). Surface bound bIgE-Fc2–4 was detected with SA-647 (top) and surface bound FcεRIα-Ova was detected with anti-Ova FITC (bottom). Source data are provided as a Source Data file.
