Concentrations of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) were measured in blood obtained from pregnant women to elucidate the fluctuations in the fibrinolytic system which occur during the course of pregnancy. The t-PA activity was measured with a modified bioimmunoassay using anti-t-PA monoclonal antibody (SP-322) against a single chain of recombinant t-PA. The t-PA antigen was measured by ELISA using the same antibody. PAI activity was determined with a competitive inhibition assay of t-PA activity. In early pregnancy, t-PA activity was found to be close to the standard range seen in nonpregnant women, and gradually decreased during the course of pregnancy, then recovered to rise to the normal range within 48 hours after delivery. The t-PA antigen and PAI activity levels rose slowly during the course of pregnancy, and fell promptly after delivery. t-PA activity and t-PA antigen in levels in umbilical cord blood were higher after vaginal delivery than after cesarean section. These findings suggest that there may be an important physiological balance of the fibrinolytic system between mother and fetus during the course of pregnancy and the puerperium.