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Related Experiment Videos

Determination of complement activating circulating immune complexes and an example for antigen specific immune

F Krapf, C Fuchs, M Herrmann

    Journal of Clinical & Laboratory Immunology
    |December 1, 1986
    PubMed
    Summary

    A new fluorescence assay effectively detects complement activating circulating immune complexes (CIC) using C3 and C4 proteins. This method shows clear differences between patients with inflammatory diseases and healthy individuals, proving its suitability for routine diagnostics.

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    Area of Science:

    • Immunology
    • Biochemistry
    • Clinical Chemistry

    Background:

    • Circulating immune complexes (CIC) play a role in various inflammatory diseases.
    • Accurate detection of CIC is crucial for diagnosis and monitoring.
    • Existing methods for CIC detection may have limitations in sensitivity or specificity.

    Purpose of the Study:

    • To develop and validate a novel fluorescence-linked immunosorbent assay (FLISA) for screening complement activating CIC.
    • To assess the assay's performance in differentiating between patients with chronic inflammatory diseases and healthy controls.
    • To explore the adaptability of the assay for antigen-specific CIC detection.

    Main Methods:

    • Developed a FLISA by coupling anti-human C3 or C4 F(ab')2 fragments to a nitrocellulose matrix.

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  • Incubated nitrocellulose-bound antibodies with serum samples, followed by reaction with fluorescence-labeled anti-human IgG or IgM.
  • Standardized the assay using a WHO Tetanus toxoid anti-Tetanus immune complex reference standard and validated with patient and control samples.

    • Main Results:

    • The FLISA demonstrated reliable standardization and reproducibility.
    • Determination of C3-IgG and C3-IgM aggregates showed the best discrimination between patients and controls.
    • The assay proved suitable for routine use due to its ease of performance and stability.
    • An antigen-specific CIC test (CEA-anti-CEA) was successfully developed using a modified system.

    Conclusions:

    • The developed FLISA is a robust and sensitive method for detecting complement activating CIC.
    • The assay effectively distinguishes patients with chronic inflammatory conditions from healthy individuals.
    • The FLISA system is versatile and can be adapted for antigen-specific CIC detection, aiding in disease-specific diagnostics.