Purification, Characterization, and Attempts at Isotopic Labeling of Mouse Interferon (original) (raw)

• Journal List
• J Virol
• v.5(2); 1970 Feb
• PMC375980

J Virol. 1970 Feb; 5(2): 145–152.

Virus Laboratory of Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19146

1 Fellow of the Rockefeller Foundation. Present address: Microbiologia, Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais, Belo Horizonte, M. G., Brazil.

2 On leave of absence from the Institute of Virology, Czechoslovak academy of Sciences, Bratislava, Czechoslovakia.

Abstract

Under optimal conditions which minimized the accumulation of extraneous proteins, interferon preparations were obtained in L cells containing from 2 × 104 to 5 × 104 units/mg of protein. The radiolabeled proteins were liberated simultaneously with interferon from cultures exposed to tritiated amino acids after viral stimulation and from corresponding controls, and were subsequently purified with the following results. Chromatography of interferon on carboxymethyl-Sephadex C-25 eliminated selectively unlabeled or poorly labeled proteins, resulting in a greater than sixfold increase in counts per minute per milligram of protein. Similarly purified control material harbored at least 12 times less tritium per milligram of protein than interferon, and the label was more diversely distributed among proteins of different molecular weights. On electrophoresis of interferon in polyacrylamide gels, labeled proteins were reduced further by a factor of at least 10 without loss in titer. Final purification was estimated at greater than 280-fold, representing a calculated specific activity of at least 1.4 × 107 units of interferon per milligram of protein.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.2M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

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

• Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. [PMC free article] [PubMed] [Google Scholar]
• CANTELL K, PAUCKER K. QUANTITATIVE STUDIES ON VIRAL INTERFERENCE IN SUSPENDED L CELLS. IV. PRODUCTION AND ASSAY OF INTERFERON. Virology. 1963 Sep;21:11–21. [PubMed] [Google Scholar]
• Fantes KH. Further purification of chick interferon. Nature. 1965 Sep 18;207(5003):1298–1298. [PubMed] [Google Scholar]
• Fantes KH. Chick interferon: heterogeneity of electric charge. Science. 1969 Mar 14;163(3872):1198–1199. [PubMed] [Google Scholar]
• Fantes KH, Furminger IG. Polyacrylamide gel electrophoresis of highly purified chick interferon. Nature. 1967 Oct 7;216(5110):71–72. [PubMed] [Google Scholar]
• Hallum JV, Youngner JS, Stinebring WR. Interferon activity associated with high molecular weight proteins in the circulation of mice injected with endotoxin or bacteria. Virology. 1965 Nov;27(3):429–431. [PubMed] [Google Scholar]
• HO M. IDENTIFICATION AND "INDUCTION" OF INTERFERON. Bacteriol Rev. 1964 Dec;28:367–381. [PMC free article] [PubMed] [Google Scholar]
• KREUZ LE, LEVY AH. PHYSICAL PROPERTIES OF CHICK INTERFERON. J Bacteriol. 1965 Feb;89:462–469. [PMC free article] [PubMed] [Google Scholar]
• LAMPSON GP, TYTELL AA, NEMES MM, HILLEMAN MR. Purification and characterization of chick embryo interferon. Proc Soc Exp Biol Med. 1963 Feb;112:468–478. [PubMed] [Google Scholar]
• LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
• MERIGAN TC. PURIFIED INTERFERONS: PHYSICAL PROPERTIES AND SPECIES SPECIFICITY. Science. 1964 Aug 21;145(3634):811–813. [PubMed] [Google Scholar]
• PAUCKER K, SKURSKA Z, HENLE W. Quantitative studies on viral interference in suspended L cells. I. Growth characteristics and interfering activities of vesicular stomatitis, Newcastle disease, and influenza A viruses. Virology. 1962 Jun;17:301–311. [PubMed] [Google Scholar]
• PAUCKER K, CANTELL K, HENLE W. Quantitative studies on viral interference in suspended L cells. III. Effect of interfering viruses and interferon on the growth rate of cells. Virology. 1962 Jun;17:324–334. [PubMed] [Google Scholar]
• ROTEM Z, CHARLWOOD PA. Molecular weights of interferons from different animal species. Nature. 1963 Jun 15;198:1066–1068. [PubMed] [Google Scholar]
• Schonne E. Properties of rat-tumor interferon. Biochim Biophys Acta. 1966 Feb 28;115(2):429–439. [PubMed] [Google Scholar]
• SIMINOVITCH L, GRAHAM AF, LESLEY SM, NEVILL A. Propagation of L strain mouse cells in suspension. Exp Cell Res. 1957 Apr;12(2):299–308. [PubMed] [Google Scholar]
• VILCEK J, TOMISOVA J, SOKOL F, HANA L. CONCENTRATION AND PARTIAL PURIFICATION OF INTERFERON FROM MOUSE BRAINS. Acta Virol. 1964 Jan;8:76–79. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)