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Sensitive radiochemical esterolytic assays for urokinase

T Imanari, G M Wilcox, J J Pisano

    Clinica Chimica Acta; International Journal of Clinical Chemistry
    |September 6, 1976
    PubMed
    Summary
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    Two new radiochemical assays simplify urokinase measurement. These sensitive methods accurately quantify urokinase activity by measuring hydrolyzed ester substrates, aligning with traditional fibrin plate assays.

    Area of Science:

    • Biochemistry
    • Enzymology
    • Biochemical Assays

    Background:

    • Urokinase is a key enzyme in fibrinolysis.
    • Accurate measurement of urokinase activity is crucial for clinical and research applications.
    • Existing assays can be complex or lack sensitivity.

    Purpose of the Study:

    • To develop and validate two novel, sensitive radiochemical esterolytic assays for urokinase.
    • To compare the performance of these new assays with the established fibrin plate assay.

    Main Methods:

    • Hydrolysis of Nalpha-acetyl-glycyl-L-lysine [3H]methyl ester by urokinase.
    • Urokinase-dependent activation of plasminogen and subsequent plasmin assay using Nalpha-tosyl-L-arginine [3H]methyl ester.
    • Extraction and scintillation counting of generated [3H]methanol.

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    Main Results:

    • Both assays demonstrated high sensitivity for urokinase detection.
    • The radiochemical assays provided results in good agreement with the fibrin plate assay.
    • The method allows for facile separation of the radiolabeled alcohol product.

    Conclusions:

    • The developed radiochemical assays offer a simple, sensitive, and reliable method for quantifying urokinase activity.
    • These assays can serve as valuable alternatives to existing methods in research and potentially clinical settings.