Figure 4.

Purification and characterization of representative evasins from three genera. For evasins, ACA-01 from A. cajennense (A), RPU-01 from R. pulchellus (B), IRI-01 from I. ricinus (C), and IHO-01 from I. holocyclus (D) are shown. Top left, the size exclusion chromatogram for purification of the evasin protein (with a C-terminal His₆ tag); bottom left, non-reducing SDS-PAGE of fractions (Fr.) spanning the main peak (boxed on the chromatogram); right, competitive fluorescence anisotropy curves for binding of the purified evasin to each of five CC chemokines. The molecular weight marker used for SDS-PAGE was Bio-Rad Precision Plus Protein unstained standards. Binding data represent the average ± S.E. (error bars) of values from three independent experiments, each recorded in duplicate. Solid lines are fitted binding displacement curves.