The functional activity of FVIIa variants. (A) Thrombin generation assays carried out using the following purified proteins: WT FVIIa, VA, and the mutants N301E and N301D using hemophilia A plasma (ie, plasma without functional FVIII). (B) The dose effect of FVIIa concentrations (from 0.03 to 1600 nM) for variants described in panel A. (A-B) The assay was carried out in the presence of phospholipid vesicles (4 µM) either in the presence (left) or absence (right) of TF (0.5 pM). (C) Effect of PL concentration (left, PL range: 0, 1-16 µM) and TF concentrations (right, TF range: 0, 0.1-25 pM) on thrombin generation by the FVIIa variants (FVIIa concentration: 50 nM) described in panel A.