Fig. 4
From: Directed evolution of broadly crossreactive chemokine-blocking antibodies efficacious in arthritis
Crossreactivity of engineered antibodies toward multiple CXC chemokines. a Heat map indicating the binding intensity of the engineered antibodies against 20 diverse human and murine CXC chemokines. Binding was assessed by flow cytometry using two cell-display arrangements: soluble CXC chemokine against yeast-displayed CK129 (red), CK138 (blue), and CK157 (gray) antibodies (on the left) and soluble serum albumin–antibody fusions SA129 (red), SA138 (blue), and SA157* (gray) against yeast-displayed CXC chemokines (on the right). Normalized binding/display signal intensities range from light to dark colors indicating low (0.0‒0.2) and high (0.8‒1.0) titers, respectively. b Binding isotherms of yeast-displayed CXC chemokines to soluble serum albumin–antibody fusions SA129, SA138, and SA157*. Equilibrium binding affinity (KD) values were determined only for chemokines exhibiting signals at high concentrations of soluble antibody fusions. CXC chemokines are gradient colored ranging from dark (high affinity) to light (low affinity) red (SA129), blue (SA138), and gray (SA157*). Data are presented as mean (dots) ± s.e.m. (bars). c Plot showing binding affinities of yeast-displayed CXC chemokines to SA129 (red), SA138 (blue), and SA157* (gray) antibody fusions. The indicated values are displayed as differently colored filled circles and represent the means of at least three independent experiments presented as inverse equilibrium binding constants (1/KD; M−1)
