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Studies on subtype composition in natural leukocyte interferon preparations.

H Shirono1, C Ito, J Koga

  • 1Biotechnology Research Laboratories, JCR Pharmaceuticals Co. Ltd., Kobe, Japan.

Journal of Virological Methods
|January 1, 1990
PubMed
Summary

Highly purified human leukocyte interferon was achieved using antibody affinity chromatography. This method enabled subtype analysis and demonstrated comparable purity to recombinant interferons, aiding natural interferon research.

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Area of Science:

  • Immunology
  • Biochemistry
  • Virology

Background:

  • Human leukocyte interferon (interferon-alpha) is a crucial antiviral protein.
  • Existing purification methods for leukocyte interferon are often complex and can reduce activity.
  • Recombinant interferons offer high purity but may not fully represent natural immune responses.

Purpose of the Study:

  • To develop a highly efficient purification method for human leukocyte interferon using specific antibodies.
  • To establish an enzyme immunoassay for interferon subtype analysis.
  • To compare the purity and characteristics of naturally occurring leukocyte interferon with recombinant forms.

Main Methods:

  • Generation and purification of antisera against human leukocyte interferon.
  • Affinity chromatography utilizing the purified antibodies for interferon purification.

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  • Development of an enzyme immunoassay for subtype characterization.
  • Reversed-phase high-performance liquid chromatography (RP-HPLC) for analyzing antiviral and immunoreactivity profiles.

    • Main Results:

    • Antibody affinity chromatography achieved a 10,000-fold purification of leukocyte interferon in a single step while preserving activity.
    • The enzyme immunoassay successfully enabled subtype analysis of interferon.
    • Crude and purified leukocyte interferons exhibited identical profiles in RP-HPLC, indicating high purity and integrity.
    • The purified natural interferon mixture showed comparable purity to recombinant interferons.

    Conclusions:

    • Antibody affinity chromatography is a highly effective method for purifying human leukocyte interferon.
    • This technique allows for the characterization of natural interferon subtypes.
    • The findings suggest that natural leukocyte interferon mixtures can be purified to a level comparable to recombinant interferons, facilitating further research into innate immune responses.