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A simple amidolytic method for the determination of functional active antithrombin III
Scandinavian Journal of Clinical and Laboratory Investigation
|January 1, 1976
Summary
A new amidolytic assay accurately measures active antithrombin III (AT III) levels in plasma. This simpler method, independent of heparin, offers higher accuracy than traditional clotting assays.
Area of Science:
- Biochemistry
- Clinical Chemistry
- Hematology
Background:
- Antithrombin III (AT III) is a crucial anticoagulant protein.
- Accurate measurement of functional AT III is essential for diagnosing and managing thrombotic disorders.
- Existing clotting-based methods can be complex and influenced by various factors.
Purpose of the Study:
- To describe a simple, accurate amidolytic method for determining functionally active antithrombin III concentration.
- To develop an assay that is independent of heparin and less influenced by fibrinogen.
Main Methods:
- Plasma is diluted and incubated with thrombin.
- EDTA is used to retard fibrin polymerization, preventing interference from fibrinogen.
- Polybrene is added to ensure heparin independence.
- Remaining thrombin is quantified using the chromogenic substrate benzoyl-Phe-Arg-p-NA.
Main Results:
- The described amidolytic method provides a simpler alternative to clotting assays.
- The assay demonstrates higher accuracy compared to traditional methods.
- Results from this assay show a close correlation with immunoassay measurements of antithrombin III.
Conclusions:
- This novel amidolytic assay offers a reliable and accurate method for measuring functional antithrombin III.
- The assay's simplicity and independence from heparin make it a valuable tool in clinical settings.
- The high correlation with immunoassay validates its utility for assessing AT III levels.