A Functional Plasminogen Assay Utilizing the Potentiating Effect of Fibrinogen to Correct for the Overestimation of Plasminogen in Pathological Plasma Samples

Summary

An overestimation of the plasminogen concentration occurs in patients with elevated levels of fibrin degradation products (fdp) and/or fibrinogen, when using assays based on the activation of plasminogen by streptokinase (Sk) followed by the hydrolysis of a synthetic chromogenic substrate. This source of error could be overcome by addition of fibrinogen in excess to the plasminogen assay thereby obtaining maximum stimulation of the Sk-plasminogen complex. The late degradation products of fibrinogen and fibrin, D_cate and D_dimer, respectively, had no potentiating effect on the Sk-plasminogen complex. The modified assay was designed as a manual method or for a centrifugal analyzer. The excellent reproducibility is illustrated by the coefficient of variation (CV) within series being 1.3% (n = 24) and from day to day being 1.9% (n = 20).

Keywords

Plasminogen assay - Sk-plasminogen - Fibrinogen - Fibrin degradation products - Amidolytic method

• References

• 1 Müllertz S. Formation and properties of the activator of plasminogen and of human and bovine plasmin. Biochem J 1955; 61: 424-434
• 2 Davies MC, Englert ME, De Renzo EC. Interaction of streptokinase and human plasminogen I. J Biol Chem 1964; 239: 2651-2656
• 3 McClintock DK, Bell PH. The mechanism of activation of human plasminogen by streptokinase. Biochem Biophys Res Commun 1971; 43: 694-702
• 4 Schick LA, Castellino FJ. Direct evidence for the generation of an active site in the plasminogen moiety of the streptokinase - human plasminogen activator complex. Biochem Biophys Res Commun 1974; 57: 47-54
• 5 Soria J, Soria C, Bertrand O, Dunn F, Samama M, Bachmann F. The amidolytic activity of the Sk-plasminogen complex is enhanced by a potentiator which is generated in the presence of vascular plasminogen activator - role of fibrin degradation products. Thromb Haemostas 1982; 47: 193-196
• 6 Soria C, Soria J, Dunn F, Deguchi K, Mirshahi M, Lijnen R, Samama M, Caen JP. Comparison between the potentiating effect induced by fibrin(ogen) derivatives on plasminogen activation induced by either streptokinase or tissue plasminogen activator. Haemostasis 1984; 14: 70
• 7 Friberger P. Method for the assay of plasminogen in plasma. Scand J Clin Lab Invest 1982; (Suppl. 42) suppl 162: 49-54
• 8 Deutsch DG, Mertz ET. Plasminogen: Purification from human plasma by affinity chromatography. Science 1970; 170: 1095-1096
• 9 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Arkiv Kemi 1956; 415-443
• 10 Haverkate F, Timan G, Nieuwenhuizen W. Anticlotting properties of fragments D from human fibrinogen and fibrin. Eur J Clin Invest 1979; 9: 253-255
• 11 Knös M, Friberger P. Methods for plasminogen determination in human plasma and for streptokinase standardization.. In Davidson JF, Cepelak V, Samama MM, Desnoyers PC. eds Progress in Chemical Fibrinolysis and Thrombolysis. Churchill Livingstone; London: 1979pp 154-158
• 12 Jespersen J, Sidelmann J. A study of the conditions and accuracy of the thrombin time assay of plasma fibrinogen. Acta Haematol 1982; 67: 2-7
• 13 Camiolo SM, Markus G, Evers JL, Hobika GH. Augmentation of streptokinase activator activity by fibrinogen or fibrin. Thromb Res 1980; 17: 697-706
• 14 Takada A, Mochizuki K, Takada Y. Effects of Sk-potentiator and fibrinogen on Sk-plasminogen complex in the presence of tranexamic acid. Thromb Res 1980; 19: 767-773
• 15 Takada A, Takada Y. Potentiation of the activation of Glu-plasminogen by streptokinase and urokinase in the presence of fibrinogen degradation products. Thromb Res 1982; 25: 229-235
• 16 Strickland DK, Morris JP, Castellino FJ. Enhancement of the streptokinase-catalyzed activation of human plasminogen by human fibrinogen and its plasminolysis products. Biochemistry 1982; 21: 721-728
• 17 Castellino FJ, Strickland DK, Morris JP, Smith J, Chibber B. Enhancement of streptokinase-induced activation of human plasminogen by human fibrinogen fragment D₁ . Ann NY Acad Sci 1983; 408: 595-601
• 18 Lucas MA, Fretto LJ, McKee PA. The relationship of fibrinogen structure to plasminogen and plasmin activity during fibrinolysis. Ann NY Acad Sci 1983; 408: 71-91
• 19 Soria J, Soria C, Samama M. A plasminogen assay using a chromogenic synthetic substrate: Results from clinical work and from studies of thrombolysis. In Davidson JF, Rowan RM, Samama MM, Desnoyers PC. eds Progress in Chemical Fibrinolysis and Thrombolysis London. Churchill Livingstone; 1978 pp 337-346
• 20 Latallo ZS, Teisseyre E, Lopciuk S. Assessment of plasma fibrinolytic system with use of chromogenic substrate. Haemostasis 1978; 7: 150-154
• 21 Pochron SP, Mitchell GA, Albareda I, Huseby RM, Gargiulo RJ. A fluorescent substrate assay for plasminogen. Thromb Res 1978; 13: 733-739
• 22 Philo RD, Gaffney PJ. Some observations on the assay of plasminogen using streptokinase and S-2251. Haemostasis 1982; 11: 66