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The complete purification of human leucocyte interferon
Scandinavian Journal of Immunology
|January 1, 1980
Summary
Researchers purified human leucocyte interferon (HuLeIF), identifying five distinct protein species. This breakthrough offers a highly purified interferon with significant biological activity for further study.
Area of Science:
- Biochemistry
- Immunology
- Protein Chemistry
Background:
- Human leucocyte interferon (HuLeIF) is a crucial antiviral protein.
- Previous purification methods yielded heterogeneous mixtures, limiting detailed analysis.
Purpose of the Study:
- To achieve the first successful purification of HuLeIF.
- To characterize the molecular properties of purified HuLeIF species.
Main Methods:
- Multi-step purification including precipitation, gel filtration, and affinity chromatography (Cu-chelate, blue dextran, antibody).
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for molecular weight determination.
Main Results:
- Purified HuLeIF comprises five distinct subspecies with molecular weights of 18,400, 19,500, 20,180, 20,900, and 22,130 daltons.
- Two major species (18,400 and 20,180 Da) account for 85% of biological activity.
- Specific activity of pure HuLeIF reached 2 x 10(9) units/mg protein, with a 50% recovery and >350,000 purification factor.
Conclusions:
- Successful purification of HuLeIF was achieved, yielding highly active protein fractions.
- Characterization revealed multiple HuLeIF species, providing insights into its heterogeneity.
- The high specific activity and purification factor demonstrate the efficacy of the developed purification strategy.