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A stable, sensitive assay for human fibronectin

H D Todd-Kulikowski, R G Parsons

    Journal of Immunological Methods
    |January 1, 1981
    PubMed
    Summary
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    A new solid-phase radioimmunoassay accurately quantifies human fibronectin. This sensitive, rapid assay is excellent for measuring fibronectin in both plasma and cellular samples.

    Area of Science:

    • Biochemistry
    • Immunology
    • Assay Development

    Background:

    • Fibronectin is a crucial extracellular matrix protein involved in cell adhesion, migration, and wound healing.
    • Accurate quantification of fibronectin is essential for understanding its role in various physiological and pathological processes.

    Purpose of the Study:

    • To develop and validate a novel solid-phase radioimmunoassay (RIA) for the sensitive and reliable quantification of human fibronectin.
    • To assess the performance characteristics of the developed RIA, including sensitivity, speed, reagent stability, and reproducibility.

    Main Methods:

    • Development of a solid-phase RIA using 125I-labeled affinity purified IgG against human plasma fibronectin.
    • Validation of the assay by comparing its sensitivity to conventional radioimmunoassays.

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  • Assessment of assay speed, reagent stability at 4°C, and reproducibility.

    • Main Results:

    • The developed RIA demonstrated high sensitivity, with a detection limit of approximately 0.5 ng, comparable to conventional RIAs.
    • The assay is rapid, completed in less than 24 hours.
    • Reagents exhibited excellent stability at 4°C for over 2 months, and the assay showed excellent reproducibility.
    • Both human plasma fibronectin and cellular fibronectin reacted equivalently in the assay.

    Conclusions:

    • A robust and sensitive solid-phase RIA for human fibronectin has been successfully developed.
    • This assay offers advantages in terms of speed, reagent stability, and reproducibility, making it a valuable tool for fibronectin quantification.
    • The assay's ability to detect both plasma and cellular fibronectin equivalently broadens its applicability in research and diagnostics.