Fixaβ

Recombinant FVIII (Kogenate™) was a generous gift from Dr. Lisa Regan of Bayer Corporation (Berkeley, CA). Dioleoyl phospholipids [Phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS)] were purchased from Avanti Polar Lipids (Alabaster, AL). The reagents α-thrombin, FVIIa, FIXaβ, FX, and FXa (Enzyme Research Laboratories, South Bend, IN), hirudin (DiaPharma, West Chester, OH), the chromogenic FXa substrate, Pefachrome Xa (Pefa-5523, CH₃OCO-D-CHA-Gly-Arg-pNA·AcOH; Centerchem Inc. Norwalk CT), Enhanced Chemifluorescence reagent (GE Healthcare Bioscience, Piscataway, NJ), recombinant human tissue factor (rTF: Innovin, Dade Behring, Deerfield, IL), flourogenic substrate (Z-Gly-Gly-Arg-AMC: Calbiochem, San Diego, CA), and thrombin calibrator (Diagnostica Stago, Parsippany, NJ) were purchased from the indicated vendors.

2D2 anti-A3 monoclonal antibody was a gift from Dr. Lisa Regan of Bayer Corporation (Berkeley, CA). Dioleoyl phospholipids phosphatidylserine/phosphatidylcholine/phosphatidylethanolamine (PSPCPE) were purchased from Avanti Polar Lipids (Alabaster, AL) and prepared as described [26 (link)]. The reagents FVIII antibody R8B12 ((GMA012) Green Mountain Antibodies, Burlington, VT), α-thrombin, FVIIa, FIXaβ, FX, and FXa (Enzyme Research Laboratories, South Bend, IN), hirudin and chromogenic FXa substrate, Chromogenix S-2765, N-a-Z-D-Arg-Gly-Arg-pNA·2HCl (DiaPharma Group, Inc, West Chester, OH), Enhanced Chemifluorescence (ECF) reagent (GE Healthcare Bioscience, Piscataway, NJ), recombinant human tissue factor (rTF), thrombin fluorogenic substrate, Z-Gly-Gly-Arg-AMC (Calbiochem, San Diego, CA), and thrombin calibrator (Diagnostica Stago, Parsippany, NJ) were purchased from indicated vendors.