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Related Experiment Videos

A sensitive and quantitative fluorescence assay for cell surface antigens

T J Higgins, H C O'Neill, C R Parish

    Journal of Immunological Methods
    |January 1, 1981
    PubMed
    Summary

    This study introduces a sensitive fluorescent sphere assay for detecting cell surface antigens. This objective method uses a standard fluorometer and offers improved sensitivity over existing techniques.

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    Area of Science:

    • Immunology
    • Biochemistry
    • Cell Biology

    Background:

    • Accurate detection of cell surface antigens is crucial for immunological research and diagnostics.
    • Existing methods like complement lysis and radioimmune assays have limitations in sensitivity or complexity.

    Purpose of the Study:

    • To describe a novel, highly sensitive assay for quantifying cell surface antigens.
    • To present a simple, objective, and cost-effective alternative to current antigen detection methods.

    Main Methods:

    • Development of a fluorescent sphere assay utilizing immunospheres coupled with antibodies or Protein A.
    • Application of the assay to detect specific antibodies bound to target cells.
    • Quantification of fluorescence using a standard laboratory fluorometer.

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    Main Results:

    • The fluorescent sphere assay demonstrated 16- to 128-fold greater sensitivity compared to complement mediated lysis and Protein A radioimmune assays.
    • The assay exhibited sensitivity comparable to rosetting assays.
    • The procedure is simple, uses commercially available reagents, and provides objective fluorescence measurements.

    Conclusions:

    • The fluorescent sphere assay is a highly sensitive and objective method for detecting cell surface antigens.
    • This assay offers a practical and improved alternative for immunological studies and diagnostics.
    • The simplicity and use of readily available reagents make this assay accessible for widespread laboratory use.