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Interferon: purification and initial characterization from human diploid cells.

E Knight

    Proceedings of the National Academy of Sciences of the United States of America
    |February 11, 1976
    PubMed
    Summary
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    Human fibroblast interferon was purified and found to be a single glycoprotein component. This purified interferon exhibits antiviral activity, confirming its identity and potential therapeutic applications.

    Area of Science:

    • Biochemistry
    • Immunology
    • Cell Biology

    Background:

    • Interferons (IFNs) are crucial signaling proteins in the innate immune system.
    • Human fibroblast interferons play a role in antiviral defense.
    • Understanding the molecular composition of IFNs is essential for their therapeutic development.

    Purpose of the Study:

    • To purify and characterize the molecular properties of human fibroblast interferon.
    • To determine the molecular weight and composition of the active interferon component.
    • To investigate whether human fibroblast interferon is a glycoprotein.

    Main Methods:

    • Purification of interferon from human diploid fibroblast cell cultures using biochemical techniques.
    • Analysis of purified interferon by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

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  • Chemical analysis involving periodic acid oxidation and Fuchsin base staining.

    • Main Results:

    • Interferon was purified approximately 5000-fold.
    • SDS-PAGE revealed a single polypeptide component with a molecular weight of 20,000.
    • The interferon activity co-migrated with this polypeptide, indicating their identity.
    • Chemical analysis suggested the presence of carbohydrates, identifying the interferon as a glycoprotein.

    Conclusions:

    • Human fibroblast interferon is a single polypeptide of 20,000 molecular weight.
    • The purified interferon is a glycoprotein, containing carbohydrate moieties.
    • This characterization provides fundamental insights into the structure of human fibroblast interferon.