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Inhibitor binding assay for angiotensin-converting enzyme.

F Fyhrquist, I Tikkanen, C Grönhagen-Riska

    Clinical Chemistry
    |May 1, 1984
    PubMed
    Summary

    A new enzyme inhibitor binding assay accurately measures angiotensin-converting enzyme (ACE) in human serum. This sensitive and specific method offers a reliable alternative for clinical diagnostics and research.

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    Area of Science:

    • Biochemistry
    • Enzymology
    • Clinical Chemistry

    Background:

    • Angiotensin-converting enzyme (ACE) plays a crucial role in the renin-angiotensin system.
    • Accurate measurement of ACE activity in biological samples is important for diagnosing and monitoring various conditions.
    • Existing methods for ACE determination may have limitations in sensitivity, specificity, or practicality.

    Purpose of the Study:

    • To introduce a novel enzyme inhibitor binding assay for quantifying ACE activity.
    • To validate the assay's performance characteristics, including sensitivity, precision, and correlation with existing methods.
    • To demonstrate the assay's applicability to human serum samples.

    Main Methods:

    • Utilized a radio-labeled specific inhibitor (125I-labeled 351A) that binds to the active site of ACE.
    • Incubated serum samples with the labeled inhibitor under controlled conditions (pH 7.0, 37°C, 2h).
    • Separated bound inhibitor via charcoal adsorption, measured radioactivity in the supernatant, and calculated ACE concentration from a standard curve.

    Main Results:

    • Achieved a sensitivity of 200 U/L for ACE in serum, equivalent to 5.0 ng of purified human lung ACE.
    • Demonstrated high precision with coefficients of variation of 3.9% (within-assay) and 6.4% (between-assay).
    • Showed excellent correlation (r = 0.98) with a spectrophotometric method in 59 samples from healthy individuals and patients.

    Conclusions:

    • The described enzyme inhibitor binding assay is a simple, specific, and sensitive method for ACE determination.
    • The assay principle is adaptable for analyzing various biological fluids, tissues, and other enzymes.
    • This novel assay offers a valuable tool for clinical diagnostics and biochemical research.

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