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Antifactor Xa activity measured with amidolytic methods.

O R Odegård, M Lie, U Abildgaard

    Haemostasis
    |January 1, 1976
    PubMed
    Summary
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    This study presents two methods for assaying antifactor Xa activity, with and without heparin. The findings reveal that antithrombin III is the primary inhibitor of factor Xa in plasma.

    Area of Science:

    • Coagulation and Thrombosis Research
    • Biochemical Assays
    • Hemostasis and Thrombosis

    Background:

    • Assessing antifactor Xa activity is crucial for understanding coagulation disorders.
    • Heparin cofactor activity, primarily mediated by antithrombin III, plays a significant role in inhibiting factor Xa.
    • Existing methods may not fully differentiate between heparin-dependent and independent inhibition of factor Xa.

    Purpose of the Study:

    • To develop and validate methods for measuring antifactor Xa activity in plasma.
    • To investigate the contribution of antithrombin III to factor Xa inactivation.
    • To correlate antifactor Xa activity with thrombin-inactivating capacity.

    Main Methods:

    • Developed two methods to assay antifactor Xa activity: Method I (with heparin) and Method II (without heparin).

    Related Experiment Videos

  • Utilized a chromogenic substrate (Bz-Ile-Glu-Gly-Arg-pNA) to measure remaining activated factor X (Xa).
  • Employed plasma with antithrombin III removed by immunoadsorption to determine inhibitor contributions.

    • Main Results:

    • Method I, measuring total Xa inactivation in the presence of heparin, strongly correlated with thrombin-inactivating capacity (r=0.85), reflecting antithrombin III levels.
    • Method II, measuring initial Xa inactivation without heparin, showed a weaker correlation (r=0.58), indicating contributions from non-antithrombin III inhibitors.
    • Antithrombin III was found to account for approximately 80% of the initial factor Xa inactivation in plasma.

    Conclusions:

    • The developed assay methods effectively measure antifactor Xa activity.
    • Antithrombin III is the principal inhibitor responsible for the initial inactivation of factor Xa in plasma.
    • The assays are robust and not significantly affected by common variations in plasma components like fibrinogen or heparin.