FIG. 3.
Fiber-binding properties in vitro tested by ELISA. (A) Wells were coated with anti-fiber tail monoclonal antibody 4D2.5, WT or R7-RGD fiber was added (amount of fiber protein in a total volume of 100 μl), and detection was performed using antiknob blocking antibody 1D6.14. (B) Wells were coated with αvβ3 integrin, WT or R7-RGD fiber was added, and integrin-bound fiber was detected using 4D2.5 antibody. (C) Wells were coated with αvβ3 integrin (1 μg of protein per well), followed by stepwise addition of recombinant penton base (Pb) protein (WT or R340E mutant) and R7-RGD fiber (1 μg of fiber protein per well in a total volume of 200 μl). The stoichiometric ratio of R7-RGD fiber to penton base molecules was 1:1, 1:5, and 1:10, indicated under the x axis as x1, x5, and x10, respectively. Results shown represent the mean of three separate experiments, ± SD.