On the Reliability of the Use of Heparin Immobilized on Agarose for the Study of the Interactions among Heparin, Thrombin and Antithrombin

Wayne W Fish; Ingemar Björk

doi:10.1055/s-0038-1657222

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1982; 48(01): 084-086
DOI: 10.1055/s-0038-1657222

Original Article

Schattauer GmbH Stuttgart

On the Reliability of the Use of Heparin Immobilized on Agarose for the Study of the Interactions among Heparin, Thrombin and Antithrombin

Authors

Wayne W Fish

The Dept. of Biochemistry, Medical University of South Carolina, Charleston, South Carolina, U.S.A. and the Dept. of Medical and Physiological Chemistry, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Biomedical Center, Uppsala, Sweden
Ingemar Björk

The Dept. of Biochemistry, Medical University of South Carolina, Charleston, South Carolina, U.S.A. and the Dept. of Medical and Physiological Chemistry, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Biomedical Center, Uppsala, Sweden

Abstract

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Summary

The extent of inhibition of thrombin was re-examined as a consequence of the sequence of addition of thrombin and antithrombin III to a column of heparin immobilized on agarose. With the use of pure enzyme, pure inhibitor, and a highly sensitive chromogenic substrate, no appreciable difference in the extent of thrombin inhibition was observed between the two sequences of addition. These observations, together with a demonstrated sensitivity of the method to variations in experimental protocol, challenge the conclusions reached in an earlier work (Hatton and Regoeczi, Thromb. Res. 1977; 10:645) which utilized this experimental approach but which employed larger quantities of reactants and a less sensitive substrate.

Keywords

Heparin - Antithrombin III - Thrombin

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References

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