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A solid-phase fluoroimmunoassay for human IgE.

G C Blanchard, R E Gardner

    Journal of Immunological Methods
    |January 1, 1982
    PubMed
    Summary
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    A new fluoroimmunoassay (FIA) accurately measures immunoglobulin E (IgE) using a sandwich technique. This method shows good correlation with radioimmunoassay, offering a reliable alternative for IgE quantification.

    Area of Science:

    • Immunology
    • Biochemistry
    • Assay Development

    Background:

    • Immunoglobulin E (IgE) plays a crucial role in allergic reactions.
    • Accurate quantification of IgE is essential for diagnosing and monitoring allergic diseases.
    • Existing methods like radioimmunoassay have limitations, necessitating alternative approaches.

    Purpose of the Study:

    • To describe a novel fluoroimmunoassay (FIA) for the quantitative measurement of human IgE.
    • To evaluate the performance characteristics of the developed FIA method.
    • To compare the results obtained by FIA with those from established radioimmunoassay techniques.

    Main Methods:

    • Development of a sandwich fluoroimmunoassay utilizing antibody-coated StiQTM samplers.
    • Specific binding of serum IgE to immobilized antibodies, followed by washing steps.

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  • Detection of bound IgE using fluorescein-conjugated antisera and measurement of fluorescence intensity via FIAX fluorometer.

    • Main Results:

    • The FIA method demonstrated assay standards ranging from 2 to 400 IU/ml.
    • Intra-assay coefficients of variation (CV) ranged from 3.3% to 8.4%, and inter-assay CV from 6.2% to 16.1%.
    • IgE concentrations measured by FIA in 74 serum samples showed good correlation with radioimmunoassay results.

    Conclusions:

    • The described fluoroimmunoassay provides a sensitive and specific method for IgE measurement.
    • The FIA method offers a reliable alternative to radioimmunoassay for clinical diagnostics.
    • Further validation may be warranted, particularly at lower IgE concentrations where precision is reduced.