Binding of VWF to the C-terminal fragments of ADAMTS13 under flow. Purified VWF in HBS-T (0 to 500 μg/mL or 0 to 2000 nM) was injected at 20 μL/min for 3 minutes over the CM5 surface covalently coupled to CUB (A), T5-8CUB (B), or T2-8CUB (C). After equilibrium was established, the HBS-T was injected to allow the dissociation phase to be recorded. The K_D was determined similarly as described in “Materials and methods.” The entries in panel D represent the means ± SD of 4 repeats (n = 4).