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Solid-phase Clq-binding fluorescence immunoassay for detection of circulating immune complexes.

M M Collins, C H Casavant, D P Stites

    Journal of Clinical Microbiology
    |March 1, 1982
    PubMed
    Summary
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    A new fluorescence immunoassay detects immune complexes using solid-phase C1q. This method is reproducible and sensitive, offering a potential alternative to radioimmunoassays for detecting immune complexes in patient serum.

    Area of Science:

    • Immunology
    • Biochemistry
    • Analytical Chemistry

    Background:

    • Immune complexes play a role in various autoimmune diseases.
    • Accurate detection of immune complexes is crucial for diagnosis and monitoring.
    • Current methods like radioimmunoassays have limitations.

    Purpose of the Study:

    • To develop and validate a novel fluorescence immunoassay for detecting immune complexes.
    • To assess the sensitivity, reproducibility, and potential interferences of the assay.
    • To compare the performance of the new assay with existing radioimmunoassay methods.

    Main Methods:

    • Development of a solid-phase fluorescence immunoassay utilizing C1q.
    • Standardization using human aggregated immunoglobulin G (IgG) to simulate immune complexes.

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  • Testing for interference from endogenous serum components, heparin, DNA, and lipopolysaccharides.
  • Comparative analysis with a fluid-phase C1q-binding radioimmunoassay.

    • Main Results:

    • The assay demonstrated a linear relationship between aggregated IgG concentration and fluorescence intensity.
    • It was reproducible and detected as little as 10 µg/ml of aggregated IgG in heat-inactivated serum.
    • Elevated immune complex levels were detected in 7/9 systemic lupus erythematosus patients.
    • Endogenous C1q, heparin, and DNA caused competitive inhibition, while bacterial lipopolysaccharides did not interfere.
    • Concordance with radioimmunoassay was 70% for normal/elevated immune complex levels.

    Conclusions:

    • A solid-phase fluorescence immunoassay for immune complex detection has been successfully developed.
    • The assay is sensitive, reproducible, and shows promise as an alternative to radioimmunoassays.
    • Understanding and mitigating interference from endogenous factors is important for accurate results.