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Related Experiment Videos

A method of membrane preparation for immunoassay.

M Liebert, B Ballou, R J Taylor

    Journal of Immunological Methods
    |December 17, 1985
    PubMed
    Summary

    Solid tissue membranes serve as effective solid-phase antigens for immunoassays like ELISA and RIA. This method preserves antigenicity for extended storage, aiding in monoclonal antibody screening.

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    Area of Science:

    • Biochemistry
    • Immunology
    • Biotechnology

    Background:

    • Immunoassays like ELISA and RIA are crucial for detecting antigens.
    • Solid tissues contain valuable antigens but require effective preparation for immunoassay use.

    Purpose of the Study:

    • To develop and validate a method for preparing solid tissue membranes as solid-phase antigens.
    • To assess the stability and utility of these membrane preparations in immunoassay applications.

    Main Methods:

    • Solid tissues were processed into membranes and fixed onto polylysine-primed 96-well plates.
    • Antigen preservation was evaluated using ELISA with monoclonal antibodies against protein (HLA) and carbohydrate (SSEA-1) antigens.
    • Antigen deposition levels and immunoassay performance (ELISA vs. RIA) were compared.

    Main Results:

    • Membrane preparations demonstrated preserved antigenicity for both protein and carbohydrate antigens.
    • Antigenic activity remained stable after prolonged frozen storage (months).
    • The method proved effective for both ELISA and RIA, with comparable utility.

    Conclusions:

    • Solid tissue membranes are a simple, rapid, and effective method for preparing solid-phase antigens.
    • This technique is particularly valuable for screening monoclonal antibodies.
    • The prepared antigen plates offer long-term storage stability.

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