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. 1974 Aug;71(8):3139–3142. doi: 10.1073/pnas.71.8.3139

Partial Purification of Human Interferon by Affinity Chromatography

C B Anfinsen 1,2, S Bose 1,2, L Corley 1,2, D Gurari-Rotman 1,2
PMCID: PMC388637  PMID: 4370251

Abstract

Human interferon prepared by challenge of leukocytes with Sendai virus, or of fibroblasts with double-stranded poly(inosinic acid)·poly(cytidylic acid), has been studied with respect to purification by affinity chromatography. Both leukocyte and fibroblast interferons are removed from crude tissue culture fluids by means of columns of antibody to leukocyte interferon attached to Sepharose-4B. The antibody was prepared in sheep using, as antigen, material that had been partially purified by gel filtration through Sephadex G-100 columns. Many of the impurities in the crude fibroblast interferon were presumably not recognized by the sheep antibodies induced by leukocyte interferon. Fibroblast interferon was, therefore, much more effectively purified as the result of this “common denominator” approach. The fibroblast product, in contrast to interferon from leukocytes, could only be harvested efficiently from the crude starting material when a carrier protein (bovine-serum albumin, and later, cytochrome c) was added to the eluting buffers to counteract losses, presumably due to adsorption on purification and assay equipment. Both varieties of interferon exhibit molecular weights of approximately 20,000-25,000, although association with higher molecular weight proteins occurs.

Keywords: antiviral glycoprotein, immunoabsorbance

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Armstrong J. A. Semi-micro, dye-binding assay for rabbit interferon. Appl Microbiol. 1971 Apr;21(4):723–725. doi: 10.1128/am.21.4.723-725.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axén R., Porath J., Ernback S. Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature. 1967 Jun 24;214(5095):1302–1304. doi: 10.1038/2141302a0. [DOI] [PubMed] [Google Scholar]
  3. Campbell D. H., Luescher E., Lerman L. S. Immunologic Adsorbents: I. Isolation of Antibody by Means of a Cellulose-Protein Antigen. Proc Natl Acad Sci U S A. 1951 Sep;37(9):575–578. doi: 10.1073/pnas.37.9.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cuatrecasas P. Affinity chromatography. Annu Rev Biochem. 1971;40:259–278. doi: 10.1146/annurev.bi.40.070171.001355. [DOI] [PubMed] [Google Scholar]
  5. Cuatrecasas P., Wilchek M., Anfinsen C. B. Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A. 1968 Oct;61(2):636–643. doi: 10.1073/pnas.61.2.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dorner F., Scriba M., Weil R. Interferon: evidence for its glycoprotein nature. Proc Natl Acad Sci U S A. 1973 Jul;70(7):1981–1985. doi: 10.1073/pnas.70.7.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fantes K. H., Furminger I. G. Polyacrylamide gel electrophoresis of highly purified chick interferon. Nature. 1967 Oct 7;216(5110):71–72. doi: 10.1038/216071a0. [DOI] [PubMed] [Google Scholar]
  8. LINDENMANN J., BURKE D. C., ISAACS A. Studies on the production, mode of action and properties of interferon. Br J Exp Pathol. 1957 Oct;38(5):551–562. [PMC free article] [PubMed] [Google Scholar]
  9. Lampson G. P., Tytell A. A., Nemes M. M., Hilleman M. R. Multiple molecular species of interferons of mouse and of rabbit origin. Proc Soc Exp Biol Med. 1966 Feb;121(2):377–384. doi: 10.3181/00379727-121-30783. [DOI] [PubMed] [Google Scholar]
  10. Ogburn C. A., Berg K., Paucker K. Purification of mouse interferon by affinity chromatography on anti-interferon globulin-sepharose. J Immunol. 1973 Oct;111(4):1206–1218. [PubMed] [Google Scholar]
  11. Sipe J. D., de Maeyer-Guignard J., Fauconnier B., de Maeyer E. Purification of mouse interferon by affinity chromatography on a solid-phase immunoadsorbent. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1037–1040. doi: 10.1073/pnas.70.4.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Strander H., Cantell K. Production of interferon by human leukocytes in vitro. Ann Med Exp Biol Fenn. 1966;44(2):265–273. [PubMed] [Google Scholar]
  13. Vilcek J., Havell E. A. Stabilization of interferon messenger RNA activity by treatment of cells with metabolic inhibitors and lowering of the incubation temperature. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3909–3913. doi: 10.1073/pnas.70.12.3909. [DOI] [PMC free article] [PubMed] [Google Scholar]

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