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Related Experiment Videos

Standard sera in solid-phase immunoassays.

O Mäkelä, F Péterfy

    European Journal of Immunology
    |October 1, 1983
    PubMed
    Summary

    Converting antibody titers from solid-phase immunoassays to weight concentrations using standard sera can introduce significant errors. Using two standard sera, even those targeting different antigens, can reduce these inaccuracies in antibody quantification.

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    Area of Science:

    • Immunology
    • Biochemistry
    • Assay Development

    Background:

    • Antibody titers from solid-phase immunoassays (SPIA) are commonly converted to weight unit concentrations.
    • This conversion relies on standard sera with known antibody concentrations.
    • Systematic studies validating this conversion method are lacking.

    Purpose of the Study:

    • To investigate the magnitude of errors in converting antibody titers to weight concentrations using standard sera.
    • To evaluate the impact of using homologous versus heterologous standard sera.
    • To assess the effectiveness of using single versus multiple standard sera for conversion.

    Main Methods:

    • Antibody concentrations were determined using quantitative precipitation or equilibrium dialysis.
    • Thirteen sera or ascites fluids were analyzed.
    • Solid-phase radioimmunoassay (SP-RIA) and enzyme-linked immunosorbent assays (ELISA) were employed using standard sera for conversion.

    Main Results:

    • Conventional solid-phase assay conversions using a single homologous standard serum resulted in up to fourfold errors.
    • Using a combination of two standard sera reduced these errors.
    • Using heterologous standard sera (directed to different antigens) did not introduce significantly greater errors than homologous standards.
    • Combining two heterologous reference sera further reduced errors, but requires antigen coat binding capacity checks.

    Conclusions:

    • The conversion of antibody titers from solid-phase immunoassays to weight concentrations is prone to significant errors.
    • Employing multiple standard sera, including heterologous ones, can mitigate these errors.
    • Careful validation, including checking antigen binding capacity, is crucial when using heterologous standards for accurate antibody quantification.

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