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Related Experiment Videos

Protein C: an automated activity assay.

O R Odegaard, K Try, T R Andersson

    Haemostasis
    |January 1, 1987
    PubMed
    Summary

    A novel snake venom activator, Protac, enables a rapid and automated assay for protein C (PC) in plasma. This method simplifies PC measurement and identified potential hereditary protein C deficiency cases.

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    Area of Science:

    • Biochemistry
    • Clinical Chemistry
    • Hematology

    Background:

    • Protein C is a crucial anticoagulant.
    • Accurate measurement of protein C activity is vital for diagnosing deficiencies and managing thrombotic disorders.
    • Existing assays can be complex or time-consuming.

    Purpose of the Study:

    • To develop and validate a simple, automated assay for protein C activation using the snake venom derivative Protac.
    • To assess the assay's performance, including speed, independence from protein C inhibitors, and correlation with existing methods.
    • To evaluate the assay's utility in identifying potential cases of hereditary protein C deficiency.

    Main Methods:

    • Utilized Protac, a snake venom derivative, for specific activation of protein C in citrated plasma.
    • Employed a chromogenic substrate (S-2366) for quantifying activated protein C.
    • Validated the assay's speed (activation < 120 seconds) and independence from plasma inhibitors.
    • Compared results with a standard immunoassay for protein C, assessing correlation.

    Main Results:

    • The Protac-based assay demonstrated rapid protein C activation (< 120 seconds).
    • Isolation of protein C from inhibitors was not required, simplifying the procedure.
    • Therapeutic heparin concentrations did not affect assay results.
    • A strong positive correlation (r = 0.92) was observed between the Protac assay and immunoassay methods.
    • Three potential cases of hereditary protein C deficiency were identified within a single family.

    Conclusions:

    • Protac provides a simple, rapid, and automated method for determining protein C activity in plasma.
    • The assay is reliable, correlates well with immunoassays, and is unaffected by therapeutic heparin levels.
    • This method facilitates the detection of protein C deficiency, aiding in the diagnosis of hereditary conditions.

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