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Recombinant equine interferon-beta 1: purification and preliminary characterization.

G R Adolf1, E Traxler, I Maurer-Fogy

  • 1Department of Cell Biology, Ernst-Boehringer-Institut für Arzneimittelforschung, Vienna, Austria.

Journal of Interferon Research
|June 1, 1990
PubMed
Summary

Equine interferon-beta 1 (EqIFN-beta 1) was purified and characterized. This equine-specific protein demonstrated potent antiviral activity in equine cells but not in other species, enabling the development of a novel immunoassay.

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Area of Science:

  • Biochemistry
  • Immunology
  • Veterinary Science

Background:

  • Interferons (IFNs) are crucial antiviral proteins.
  • Equine interferon-beta 1 (EqIFN-beta 1) is a specific type of IFN with potential therapeutic applications in horses.
  • Understanding the properties and species-specificity of EqIFN-beta 1 is essential for its development.

Purpose of the Study:

  • To purify and characterize recombinant Equine interferon-beta 1 (EqIFN-beta 1).
  • To assess the antiviral activity and species-specificity of EqIFN-beta 1.
  • To develop tools for the detection and quantification of EqIFN-beta 1.

Main Methods:

  • Purification of EqIFN-beta 1 from recombinant Escherichia coli using sequential chromatography (hydroxylapatite, anion-, and cation-exchangers).

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  • Purity assessment via SDS-PAGE, gel permeation HPLC, and reverse-phase HPLC.
  • Amino-terminal amino acid sequencing.
  • Antiviral activity assays using various cell lines and viruses.
  • Development of monoclonal antibodies and a solid-phase enzyme immunoassay.

    • Main Results:

    • EqIFN-beta 1 was purified to >98% purity.
    • The protein primarily contained an additional amino-terminal methionine.
    • Specific antiviral activity was high (5 X 10(8) U/mg) in equine cells but negligible (<0.001%) in human, murine, ovine, and bovine cells.
    • Monoclonal antibodies neutralized EqIFN-beta 1 but not human IFN-beta, and vice versa.
    • A rapid one-step solid-phase enzyme immunoassay for EqIFN-beta 1 was established.

    Conclusions:

    • Recombinant EqIFN-beta 1 is a highly pure and potent antiviral agent specific to equine cells.
    • The developed monoclonal antibodies and immunoassay are valuable tools for studying and utilizing EqIFN-beta 1.