Abstract
The clotting time (CT) of fibrinogen mixed with thrombin decreased, then increased with increasing fibrinogen levels. By contrast, log CT decreased monotonically with respect to the log level of activating enzyme (thrombin or reptilase). Here, the CT was determined over a large range of fibrinogen concentration (to 100 mg ml(-1)) at a fixed level of enzyme. A new parameter, [Fib]min, the minimal fibrinogen concentration required for thrombin or reptilase-instigated phase change (coagulation), was determined as [Fib]min = 0.2 +/- 0.05 microM fibrinogen. A dynamic simulation program (Stella) was employed to organize simulations based on simple and complex coagulation mechanisms, which generated CT values. The successful simulation aimed at forming [Fib]min and "recognized" the binding of unreacted fibrinogen with intermediate fibrin protofibrils. The "virtual data" mimicked the biphasic experimental CT values over a wide range of concentrations. Fibrinogen appeared to act in three modalities: as a thrombin substrate; as a precursor of fibrin; and as a competitor for fibrin protofibrils. The optimized simulation may provide a basis for predicting CT in more complex systems, such as pathological plasmas or whole blood or at high concentrations encountered with fibrin sealant.