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Simple immunochromometric assay for protein C activity.

T Exner, J Koutts

    The Journal of Laboratory and Clinical Medicine
    |May 1, 1986
    PubMed
    Summary
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    This study introduces an immunochromometric assay (ICMA) for quantifying activated protein C (APC) in plasma. The assay effectively detects inactive protein C forms in patients on anticoagulants, improving diagnostic capabilities.

    Area of Science:

    • Biochemistry
    • Immunology
    • Clinical Chemistry

    Background:

    • Protein C is a crucial anticoagulant protein.
    • Existing methods like IRMA may not fully distinguish between active and inactive protein C forms.
    • Accurate quantification of activated protein C is vital for understanding coagulation disorders.

    Purpose of the Study:

    • To develop and validate an immunochromometric assay (ICMA) for quantifying activated protein C (APC).
    • To compare the ICMA with an immunoradiometric assay (IRMA) for protein C antigen detection.
    • To assess the assay's utility in identifying inactive protein C forms.

    Main Methods:

    • Utilized antibody-coated wells for protein C capture.
    • Employed a heterologous antibody for detection in the ICMA.

    Related Experiment Videos

  • Activated protein C with bovine thrombin and quantified using chromogenic substrates (S-2366 or CBS 34.47).
  • Performed assays in a 96-well microtiter plate system.

    • Main Results:

    • The ICMA successfully quantifies activated protein C.
    • Good correlation was observed between IRMA and ICMA for protein C antigen in normal individuals and liver disease patients.
    • Patients on oral anticoagulants showed reduced protein C antigen by IRMA but significantly lower levels by ICMA, indicating the presence of inactive protein C.

    Conclusions:

    • The developed ICMA is a reliable method for measuring activated protein C.
    • The assay can differentiate between active and inactive protein C forms.
    • ICMA provides valuable insights into protein C status, particularly in patients with altered coagulation profiles.