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Solubilization of human thyroid microsomal antigen.

S Mariotti, A Pinchera, C Marcocci

    The Journal of Clinical Endocrinology and Metabolism
    |February 1, 1979
    PubMed
    Summary
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    Researchers investigated methods to solubilize human thyroid microsomal antigen. Triton X-100 proved most effective, enabling antigen purification and identification for potential diagnostic applications.

    Area of Science:

    • Immunology
    • Biochemistry
    • Endocrinology

    Background:

    • Human thyroid microsomal antigen is crucial in autoimmune thyroid diseases.
    • Effective solubilization is necessary for antigen purification and characterization.

    Purpose of the Study:

    • To evaluate various agents for solubilizing human thyroid microsomal antigen.
    • To assess the effectiveness of detergents, salts, and enzymes in antigen release.
    • To establish a basis for antigen identification and purification.

    Main Methods:

    • Solubilization using Triton X-100, deoxycholate, 3 M KCl, papain, and trypsin.
    • Separation of solubilized antigen by high-speed centrifugation.
    • Measurement of antigenic activity using 125I-labeled anti-microsomal IgG (anti-M IgG).

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  • Affinity chromatography for antigen purification.

    • Main Results:

    • Triton X-100 was the most effective solubilizing agent, followed by deoxycholate, papain, trypsin, and 3 M KCl.
    • Solubilized materials specifically bound and inhibited anti-M IgG binding to microsomes.
    • Affinity chromatography yielded a 15.6-fold purification of anti-M antibodies.

    Conclusions:

    • Multiple agents can effectively solubilize human thyroid microsomal antigen.
    • Triton X-100 demonstrates superior efficacy in antigen solubilization.
    • These methods provide a foundation for the purification and identification of thyroid microsomal antigen.