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Related Experiment Videos

Lymphocyte surface membrane immunoglobulin

R J Winchester, S M Fu

    Scandinavian Journal of Immunology
    |June 1, 1976
    PubMed
    Summary
    This summary is machine-generated.

    False positive surface immunoglobulin detection on lymphocytes can be avoided. Using F(ab)2 antibody fragments prevents immune complex formation, leading to more accurate results in flow cytometry analysis.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Biotechnology

    Background:

    • Surface immunoglobulin (Ig) detection on lymphocytes is crucial for cell identification and function analysis.
    • Immune complex formation between fluorochrome-tagged antibodies and residual serum Ig can interfere with accurate surface Ig detection.
    • This interference leads to falsely elevated percentages of Ig-bearing cells due to binding to Fc receptors.

    Purpose of the Study:

    • To describe methods for improving the accuracy of surface Ig detection on lymphocytes.
    • To present strategies for mitigating interference from immune complex formation in fluorescence assays.
    • To enhance the reliability of flow cytometry for analyzing Ig-positive cells.

    Main Methods:

    • Utilizing fluorochrome-tagged F(ab')2 fragments of antibodies, which lack an exposed Fc region.

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  • Describing methods for the preparation of these F(ab')2 fragments.
  • Employing latex ingestion to aid monocyte identification and facilitate the elution of autoreactive anti-lymphocyte antibodies.

    • Main Results:

    • F(ab')2 fragments form immune complexes that do not bind to Fc receptors, thus preventing false positives.
    • The use of F(ab')2 fragments significantly reduces interference from residual serum Ig.
    • Latex ingestion aids in reducing false positives by diminishing autoreactive anti-lymphocyte antibody binding.

    Conclusions:

    • The use of F(ab')2 antibody fragments is a reliable method to prevent immune complex-mediated interference in surface Ig detection.
    • These improved methods enhance the accuracy and reliability of flow cytometry for quantifying Ig-bearing lymphocytes.
    • The described techniques offer a significant advancement for immunological and cell biology research.