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Immunochemical analysis of active and inactive antithrombin III
British Journal of Haematology
|October 1, 1980
Summary
Accurate Antithrombin III (AT) measurement requires specific antisera that distinguish active AT from inactive AT/protease complexes. Plasma, not serum, is the recommended clinical test material for reliable immunoassay results.
Area of Science:
- Clinical Chemistry
- Immunology
- Hematology
Background:
- Accurate measurement of Antithrombin III (AT) is crucial for assessing coagulation disorders.
- Immunoassays for AT can yield discrepant results depending on the antiserum and sample type (plasma vs. serum).
Purpose of the Study:
- To investigate the causes of anomalous results in Antithrombin III (AT) immunoassays.
- To determine the optimal conditions for reliable AT measurement using electroimmunoassay (EIA).
Main Methods:
- Comparison of electroimmunoassay (EIA) with a chromogenic assay (S2238) for Antithrombin III (AT) measurement.
- Evaluation of different antisera specificity for active and inactive AT forms.
- Analysis of plasma and serum samples with and without added thrombin.
Main Results:
- One antiserum produced discrepant results between plasma and serum, not correlating with functional activity.
- A different antiserum successfully differentiated active AT from AT/protease complexes, showing high correlation (r=0.97) with functional activity.
- Addition of thrombin elucidated antiserum specificity issues, explaining previous anomalies in AT immunoassays.
Conclusions:
- Plasma is the preferred clinical sample for Antithrombin III (AT) testing over serum.
- Antisera used in EIA for AT must differentiate free AT from AT/protease complexes for results to correlate with functional activity.