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Related Experiment Videos

Standardized automated assay for functional alpha 1-antitrypsin.

R E Mullins, R L Miller, R L Hunter

    Clinical Chemistry
    |November 1, 1984
    PubMed
    Summary

    A new enzymic assay accurately measures alpha 1-antitrypsin functional activity using a specific trypsin substrate. This method offers improved precision and reliability for quantifying this important protein in serum samples.

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    BioMed research international·2017

    Area of Science:

    • Biochemistry
    • Enzymology
    • Clinical Chemistry

    Background:

    • Alpha 1-antitrypsin (AAT) is a crucial serine protease inhibitor in the blood.
    • Accurate measurement of AAT functional activity is vital for diagnosing AAT deficiency and related diseases.
    • Existing assays may have limitations in substrate solubility and precision.

    Purpose of the Study:

    • To develop and validate a novel enzymic assay for quantifying the functional activity of alpha 1-antitrypsin.
    • To establish a reliable and precise method for clinical laboratory use.
    • To compare the new assay's performance against established methods.

    Main Methods:

    • Utilized tosyl-Gly-Pro-Lys-p-nitroanilide acetate (Chromozym PL) as a trypsin substrate.
    • Employed p-nitrophenyl-p'-guanidinobenzoate for active site titration to standardize the assay.
    • Standardized results in terms of active trypsin sites inhibited per liter of serum.

    Main Results:

    • The assay demonstrated good agreement with immunonephelometric methods (r = 0.953).
    • Achieved high precision with within-run and run-to-run variability of approximately 3%.
    • Showed linear results for AAT concentrations ranging from 0 to 60 mumol/L, with a minimum sample volume of 50 microL.

    Conclusions:

    • The developed enzymic assay provides a precise and reliable method for measuring alpha 1-antitrypsin functional activity.
    • This assay is suitable for clinical applications due to its accuracy, precision, and efficiency.
    • The assay's performance characteristics make it a valuable tool for AAT deficiency diagnostics.

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