Figure 8.
Design and analysis of hybrid evasins. A, the construction of hybrid evasins is indicated in the diagram. B, binding affinities (Kd, m) of immobilized P672, P672_EVA1 hybrids, and EVA1 to human CC-chemokines using biolayer interferometry. High-affinity binding is indicated in shades of green, medium affinity in yellow, and low affinity in shades of orange. Chemokines are arranged by sequence similarity–based phylogeny. A dash (—) indicates that binding was not detected at 300 nm chemokine concentration. C, neutralization of CCL8-induced THP-1 monocyte cell migration by P672(1_44)-EVA1(29_94). The y axis shows cell count migrating through to the bottom chamber in response to an EC80 dose of CCL8. The Data (three technical replicates and three biological replicates) are shown as mean ± S.E. The x axis shows P672(1_44)-EVA1(29_94) concentration (log10 molar). IC50 was estimated by fitting an agonist response curve with four parameters as described (14).