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Related Experiment Videos

Naturally occurring carbohydrate antibodies: interference in solid-phase immunoassays.

R G Hamilton, N F Adkinson

    Journal of Immunological Methods
    |February 28, 1985
    PubMed
    Summary

    Interfering antibodies against carbohydrate matrices like agarose and cellulose can cause false positives in antibody assays. Pre-treating samples with agarose removes these antibodies, improving assay accuracy.

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    Area of Science:

    • Immunology
    • Biochemistry
    • Analytical Chemistry

    Background:

    • Carbohydrate matrices (agarose, cellulose) are used in protein insolubilization for antibody assays.
    • These assays can be affected by non-specific and specific interfering factors.
    • Particle-based solid-phase radioimmunoassays (SPRIAs) for antigen-specific human IgG are particularly susceptible.

    Purpose of the Study:

    • To examine factors diminishing the quality of particle-based solid-phase radioimmunoassays (SPRIAs).
    • To investigate interfering factors in antigen-specific human IgG assays using carbohydrate matrices.
    • To identify and characterize the cause of variable background binding in these assays.

    Main Methods:

    • Analysis of constant non-specific binding and absorbable binding in human and rabbit sera.

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  • Utilizing ultracentrifugation and immune-complex studies to assess IgG binding.
  • Employing inhibition assays with acid-hydrolyzed soluble agarose.
  • Performing pre-absorption of sera with agarose for SPRIAs.

    • Main Results:

    • Constant non-specific binding was consistently 1-3% Bmax across sera.
    • Absorbable binding varied widely (0.75-30% Bmax) and was linked to specific IgG antibodies against the carbohydrate matrix.
    • IgG anti-agarose antibodies were detected in a significant proportion of rabbit sera.
    • Pre-absorption with agarose removed >90% of anti-agarose antibodies, eliminating false positives.

    Conclusions:

    • Naturally occurring antibodies against carbohydrate matrices (e.g., agarose, cellulose) can interfere with solid-phase immunoassays.
    • These antibodies, often found in heterologous antisera, cause variable background binding and false positive results.
    • Pre-absorption techniques can mitigate interference from these carbohydrate antibodies, enhancing assay reliability.