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The enzymatic assay for the determination of plasminogen via the clotting time (JACOBI et al., Dtsch. med. Wochenschr. 101 (1976) 1220–1222   0.2 ml plasminogen reagent (test-kit Behringwerke/Hoechst AG)   0.1 ml plasma, diluted 1:100, incubation time 3 min. 37°C  0.05 ml thrombine (3 NIH U/ml), clotting time  is based on the following sequence of reactions: (plasminogen reagent = fibrinogen, bovine plasminogen and streptokinase, which does not react with bovine plasminogen) human plasminogen and streptokinase form an activator, which converts bovine plasminogen to bovine plasmin, which splits fibrinogen during the incubation time. The thrombine clotting time of the resulting mixture containing fibrinogen and fibrinogen degradation products depends on the amount of bovine plasmin in the assay, which is in turn a function of human plasminogen concentration. The assay is described by a kinetic model in terms of non-linear differential equations. On the basis of this model the clotting time can be computed as a function of the parameters of the assay: concentration of fibrinogen, human plasminogen, bovine plasminogen and incubation time. By computer simulation an optimal parameter set is obtained for which the resulting calibration curve is linear and not sensitive against the variances of the reagent. Furtermore, the model gives a detailed insight into the mechanism involved in the interaction of human plasminogen, fibrinogen, bovine plasminogen and thrombine.

Simulation of Coupled Enzyme Assays for a New Determination of Plasminogen Via the Clotting Time