Figure 4.
Thrombin generation potential of D519V/E665V/K1813A and D519V/E665V FVIII mutants. WT-FVIII (1 nM) or FVIII mutants (D519V/E665V; 0.125-1 nM, and D519V/E665V/K1813A; 0.125-1 nM) were added to commercial FVIII-deficient plasma. These samples were incubated with recombinant tissue factor (rTF) (1 pM) and PL vesicles (4 μM) before the addition of fluorogenic substrate and CaCl2 at the start of the assay as described in “Methods.” Experiments were performed 3 times, and the mean values are shown. Thrombin generation curves are shown (gray, FVIII-deficient plasma; orange, WT 1 nM; red, mutants 0.125 nM; green, mutants 0.25 nM; blue, mutants 0.5 nM; and light purple, mutants 1 nM).
