Figure 4.

Illustrations of SPR binding study design. A, experimental designs for cytokines binding to IL-36R, IL-1RAcP, or Fc-linked IL-36R·IL-1RAcP heterodimer. Receptors are captured onto the chip surface (yellow bar) either though biotin–NeutrAvidin interactions or through Fc–anti-Fc interactions. Cytokines or receptors are injected in solution at various concentrations. B, binding events associated with IL-1RAcP binding to IL-36R in the presence of IL-36α. Various concentrations of IL-1RAcP and a saturating concentration of IL-36α (4 μm) are added to the chip surface captured with IL-36R. Fast-on and fast-off kinetics allows IL-36α to quickly bind to IL-36R, followed by IL-1RAcP recruitment.