Extended Data Fig. 3: Characterization of Neo-4 binding properties.
From: Design of cell-type-specific hyperstable IL-4 mimetics via modular de novo scaffolds
a, Equilibrium bio-layer interferometry (BLI) titrations of hIL-4 or hNeo-4 against immobilized hIL-4Rα. b, BLI sensograms depicting interactions between hIL-4 or hNeo-4 and the hIL-4Rα/hγc complex. Equilibrium dissociation constants (KD) derived from the kinetic parameters are indicated. Equilibrium BLI titrations of hIL-4 or hNeo-4 (3-fold serial dilutions starting at 333 nM for both) against the hIL-4Rα/hγc complex are shown at right. c-d, BLI sensograms depicting the interaction between hNeo-4 or Neo-2 and immobilized (c) IL-2Rβ and (d) hIL-2Rβ/hγc complex. In (c) biosensors were exposed to 3-fold serial dilutions of Neo-2 starting at 60 nM or hNeo-4 starting at 100 nM, respectively; and in (d) biosensors were exposed to 3-fold serial dilutions of Neo-2 starting at 33 nM or hNeo-4 starting at 300 nM, respectively. KD values derived from the kinetic parameters are indicated. Equilibrium BLI titrations of hNeo-4 or Neo-2 against immobilized (c) hIL-2Rβ and (d) hIL-2Rβ/hγc are shown at right. e, Equilibrium BLI titrations of mIL-4 or mNeo-4 against immobilized mIL-4Rα extracellular domain. f, BLI sensograms depicting interactions between mIL-4 or mNeo-4 (3-fold serial dilutions starting at 650 nM for mIL-4 and 750 nM for mNeo-4) and mIL-4Rα/mγc complexes. KD values derived from the kinetic parameters are indicated. Equilibrium BLI titrations of mIL-4 or mNeo-4 against the mIL-4Rα/mγc complex are shown at right. All raw data were fitted using a 1:1 Langmuir binding model. Fitted curves are shown in gray.
