Inhibition of fVIII binding to vWf.

rfVIII at 0.2 μg/mL final concentration was mixed for 30 minutes at 37°C with different concentrations of LE2E9 before addition to vWf-coated plates. The plates were then incubated for 2 hours at RT, and the binding of fVIII was detected by addition of the HRP-labeled anti-fVIII A2 domain mAbF15B12. The mean of fVIII binding in the absence (■) or presence of antibody (○) with the standard deviation of triplicates are indicated. Controls in the absence of fVIII gave OD₄₉₀ lower than 0.05. The experiments were repeated 3 times with similar results.