Plasmin Potency Estimates: Influence of the Substrate Used in Assay

Summary A urokinase-activated plasmin (UK-plasmin) preparation was assayed against the International Reference Preparation for Plasmin (IRP-plasmin) using caseinolytic, fibrinolytic, fibrinogenolytic and chromogenic assay methods. The relative potency (using multi-dose bioassays) was estimated by the fibrinolytic method to be about twice that obtained by the caseinolytic, fibrinolytic and chromogenic assay methods. It was found that the UK-plasmin binds to fibrin to a greater extent than does the IRP-plasmin and this is advanced as an explanation for the discrepancy between assay methods. This difference in the binding of the two plasmins to fibrin may mean that it will be difficult to compare the fibrinolytic activities of various plasmin preparations. It is also shown that, during thermal degradation, the IRP-plasmin loses fibrinolytic activity more rapidly than amidolytic activity.

[1]  B. Lämmle,et al.  Different Assessment of Plasmin with Different Substrates , 1980, Thrombosis and Haemostasis.

[2]  Y. Tamaura,et al.  Fibrin Suspension as a Substrate for Plasmin: Determination and Kinetics , 1979, Thrombosis and Haemostasis.

[3]  G. Markus,et al.  Quantitative determination of the binding of epsilon-aminocaproic acid to native plasminogen. , 1978, The Journal of biological chemistry.

[4]  T. Vukovich.  Quantitative determination of plasmin: a fibrinogenolytic method. , 1977, Clinica chimica acta; international journal of clinical chemistry.

[5]  T. Kirkwood,et al.  Problems in the assay of thrombin using synthetic peptides as substrates. , 1977, Thrombosis research.

[6]  B. Wiman,et al.  The specific interaction between plasminogen and fibrin. A physiological role of the lysine binding site in plasminogen. , 1977, Thrombosis research.

[7]  G. Claeson,et al.  Methods for the Determination of Plasmin, Antiplasmin and Plasminogen by Means of the Substrate S-2251 , 1977, Thrombosis and Haemostasis.

[8]  T. Kirkwood,et al.  A Standard for Human Plasmin , 1975, Thrombosis and Haemostasis.

[9]  B. Blombäck,et al.  Prothrombin activation induced by Ecarin - a prothrombin converting enzyme from Echis carinatus venom. , 1975, Thrombosis research.

[10]  P J Campbell,et al.  International biological standards and reference preparations. I. Preparation and presentation of materials to serve as standards and reference preparations. , 1974, Journal of biological standardization.

[11]  N. Marsh,et al.  Evaluation of the Fibrin Plate Method for Estimating Plasminogen Activators , 1972, Thrombosis and Haemostasis.

[12]  H. Ekert,et al.  The Preparation and Evaluation of a Standardized Fibrin Plate for the Assessment of Fibrinolytic Activity , 1970, Thrombosis and Haemostasis.

[13]  A. J. Johnson,et al.  Assay Methods and Standard Preparations for Plasmin, Plasminogen and Urokinase in Purified Systems, 1967-1968 , 1969, Thrombosis and Haemostasis.

[14]  P. Wolf.  Modification of the Fibrin Agar Plate for Measurements of the Components of the Fibrinolytic System , 1968, Thrombosis and Haemostasis.

[15]  N. Kjeldgaard,et al.  Urokinase—an Activator of Plasminogen From Human Urine , 1961, Angiology.

[16]  M. Derechin.  The assay of human plasminogen with casein as substrate. , 1961, The Biochemical journal.

[17]  M. Lassen.  The esterase activity of the fibrinolytic system. , 1958, The Biochemical journal.

[18]  P. S. Roberts.  Measurement of the rate of plasmin action on synthetic substrates. , 1958, The Journal of biological chemistry.

[19]  N. Kjeldgaard,et al.  Urokinase an activator of plasminogen from human urine. I. Isolation and properties. , 1957, Biochimica et biophysica acta.

[20]  S. Sherry,et al.  The action of plasmin on synthetic substrates. , 1954, The Journal of biological chemistry.

[21]  A. Fletcher.  The measurement of the components of the plasminogen-plasmin system in biological fluids. , 1954, The Biochemical journal.

[22]  P. Cohen,et al.  Partial purification and properties of a proteolytic enzyme of human serum. , 1949, The Journal of biological chemistry.

[23]  P. Permin,et al.  Properties of the Fibrinokinase-Fibrinolysin System , 1947, Nature.

[24]  C. Macleod,et al.  A PROTEOLYTIC ENZYME OF SERUM: CHARACTERIZATION, ACTIVATION, AND REACTION WITH INHIBITORS , 1945, The Journal of general physiology.