A Method to Correct for the Continuing Activation during the Second Stage in a Two-Stage Assay

W Berg

doi:10.1055/s-0038-1651192

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1968; 19(01/02): 161-168
DOI: 10.1055/s-0038-1651192

Originalarbeiten – Original Articles – Travaux Originaux

Schattauer GmbH

A Method to Correct for the Continuing Activation during the Second Stage in a Two-Stage Assay

Exemplified by Urokinase Activation of Plasminogen Determined by the Lysis Time Method

Authors

W Berg

¹Medical Department II, Sahlgrenska Hospital, University of Göteborg, Sweden

Abstract

Summary

In coagulation and fibrinolysis, kinetic data are difficult to obtain with ordinary quantitative methods because no simple means are available to stop the reaction before the determination of the activity formed.

This work describes how to calculate, from the data recorded, the amount of activity at the moment the determination is started.

A formula is given for a zero, a first and a second-order reaction.

The method is exemplified by urokinase activation of plasminogen into plasmin. The determination of the plasmin activity is done by means of the lysis time method.

Full Text

References

References
1 Alkjaersig N, Fletcher A.P, Sherry S. The activation of human plasminogen. J. biol. Chem. 1958; 233: 86
2 Berg W. The urokinase activation of plasminogen and inactivation of the plasmin formed. A kinetic study. Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 19: 161
3 Berg W, Korsan-Bengtsen K, Ygge J. Plasminogen assay by means of the lysis time method. Thrombos. Diathes. haemorrh. (Stuttg.) 1965; 14: 127
4 Berg W, Korsan-Bengtsen K. Separation of human fibrinogen and plasminogen by means of gel filtration. Thrombos. Diathes. haemorrh. (Stuttg.) 1963; 9: 151
5 Berg W, Korsan-Bengtsen K, Ygge J. Preparation of human plasminogen suitable as substrate for quantitative determination of plasminogen activators. Thrombos. Diathes. haemorrh. (Stuttg.) 1966; 15: 511
6 Berg W, Korsan-Bengtsen K, Ygge J. Human and bovine plasminogen-free thrombin, purified by means of gel filtration and ion exchange chromatography. Thrombos. Diathes. haemorrh. (Stuttg.) 1966; 15: 501
7 Berg W, Korsan-Bengtsen K, Ygge J. Theoretical basis and standardization of the one-stage lysis time method for determination of urokinase. Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 19: 169
8 Derechin M. The assay of human plasminogen with casein as substrate. Biochem. J. 1961; 78: 443
9 Kjeldgaard N.O, Ploug J. Urokinase an activator of plasminogen from human urine. II Mechanism of plasminogen activation. Biochim. biophys. Acta (Amst.) 1957; 24: 283
10 Mackay M. Fractionation and standardization of plasma fibrinolytic components. Brit. med. Bull. 1964; 20: 189