Extended Data Fig. 3. Effect of the two cAbs evaluated using different assays.
a) Whole blood from healthy donors (n = 3) was incubated with FVIIa, HMB-001 or cAb as indicated in the presence of an AlexaFluor-647 labelled FVIIa-specific VHH with or without 25 mM PAR-1 AP and 1 mg/mL CRP-XL for 10 min at 37 °C. FVIIa binding to platelets was assessed with FACS. Data are expressed as mean MFI ± s.d. b) Fibrin-dependent platelet pseudoaggregation was measured in GT-like washed platelets in presence of 10 nM FVIIa, 0 or 10 nM HMB-001 and 0 or 10 nM cAb. Data are expressed as mean lag time ± s.d. (n = 3). c) Platelets were stimulated with both CRP-XL and PAR-1 AP, and thrombin generation was measured for 240 min in conditions mimicking scenario 3 (Table S8) (n = 3). FVIIa, Total FVII(a), HMB-001 or cAb was added as indicated. Bars represent mean endogenous thrombin potentials (ETP), error bars represent s.d. d) Area under the curve (AUC) of fibrin deposition in GT-like whole blood from healthy donors (n = 3) with 5 nM FVIIa and 0 or 100 nM HMB-001 or 0 or 100 nM cAb. Bars represent mean total fibrin formation in 20 min (sum of fluorescent intensity; SFI), error bars represent s.d.