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Clinical Chemistry 34: 1971-1975, 1988;
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Clinical Chemistry, Vol 34, 1971-1975, Copyright © 1988 by American Association for Clinical Chemistry

An automated approach to characterize, in simple kinetic terms, the generation and inactivation of thrombin and the interaction of fibrinogen with thrombin

DR Hoak, SK Banerjee and G Kaldor
VA Medical Center, Allen Park, MI 48101.

Here, we used a fully automated, computer-directed centrifugal analyzer (which permitted simultaneous turbidimetry and calculation of results) and purified thrombin, fibrinogen, and various inhibitors to study clot formation. The Km and Vm for these reactions were useful in detecting and partly characterizing anticoagulants. We also explored the generation and inactivation of thrombin, using the two-stage prothrombin time and antithrombin activity tests. The amount of thrombin instantaneously generated and inactivated was monitored under artificially created pathological conditions. The pseudo-first-order rate constant for thrombin generation and inactivation and the instantaneous concentration of enzymatically active and inactive thrombin were used in the characterization of these conditions. We believe this approach is suitable for routine clinical use.




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Copyright © 1988 by the American Association for Clinical Chemistry.