Human leukocyte interferon: production, purification to homogeneity, and initial characterization

Area of Science:

• Biochemistry
• Protein Chemistry
• Immunology

Background:

• Human leukocyte interferon is crucial for antiviral responses.
• Previous purification methods for interferon were complex and less efficient.
• Characterizing specific interferon species is vital for therapeutic development.

Purpose of the Study:

• To develop a novel method for protein fractionation using high-performance liquid partition chromatography (HPPLC).
• To isolate and purify a specific species of human leukocyte interferon to homogeneity.
• To characterize the purified interferon's properties and antiviral activity.

Main Methods:

• High-performance liquid partition chromatography (HPPLC) for protein separation.
• Sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) for purity assessment.
• Amino acid analysis for specific activity determination.

Main Results:

• HPPLC enabled the isolation of a homogeneous human leukocyte interferon species.
• The purified interferon showed a single sharp peak on HPPLC and SDS-PAGE.
• Antiviral activity was confirmed to coincide with the purified protein band.
• Specific activity of the pure interferon was determined to be 2-4 X 10(8) units/mg.
• The molecular weight of the purified interferon was found to be 17,500–18,000 Da.

Conclusions:

• HPPLC is an effective technique for purifying human leukocyte interferon.
• The purified interferon is homogeneous and possesses significant antiviral properties.
• This purification method facilitates further research into interferon's structure and function.