The use of von Willebrand factor [VWF antigen (WF:Ag)] measurement as a marker of endothelial cell activation for monitoring hypertensive pregnancies is limited by the poor definition of reference values. We reassessed these reference values using different assays, and those of the propeptide (VWF:Ag II) and factor VIII coagulant activity (factor VIII:C), in a large population of normal pregnancies, at 3-week intervals of gestational age. Plasminogen activator inhibitors (PAI-1 and PAI-2) were measured in parallel. Blood was collected at single time points between 12 weeks and delivery in 306 women undergoing normal singleton pregnancy. For clinical purposes, the VWF:Ag reference values were assay independent and the influence of ABO blood group on VWF:Ag or factor VIII:c was found to be limited during the third trimester. VWF:Ag II was not influenced by the ABO blood group. The ratio, VWF:Ag/factor VIII:C was close to 1.0 throughout pregnancy. In contrast, VWF:Ag II increased more slowly than VWF:Ag and the ratio of VWF:Ag II to VWF:Ag in plasma decreased from 1.00 to 0.5 at term. PAI-1 and PAI-2 increased with gestational age, but PAI-2 decreased during the last 2 weeks, indicating physiological placental regression at the very end of pregnancy.