• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Jun 1986; 83(11): 3949–3953.
PMCID: PMC323642

Purification, characterization, and antitumor activity of nonrecombinant mouse tumor necrosis factor.


Mouse tumor necrosis factor (TNF) was purified from serum through a series of steps, and each step was monitored for L-cell cytotoxicity in vitro and tumor-necrotizing activity in vivo. The two activities copurified and could not be dissociated. Purified mouse TNF has a specific activity of 2.2 X 10(7) (L-cell assay in the absence of actinomycin D) and 1 microgram causes necrosis of the standard TNF-sensitive sarcoma Meth A. TNF has a Mr of 39,000 +/- 2000 by gel filtration and a Mr of 16,000-18,000 by NaDodSO4/PAGE. Both molecular weight forms display cytotoxic and necrotizing activities. TNF has a pI of 3.9 and is destroyed by trypsin, protease, elastase, and alpha-chymotrypsin but not by neuraminidase or papain. These characteristics of nonrecombinant mouse TNF clearly resemble those of recombinant human and mouse TNF.

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 (959K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Carswell EA, Old LJ, Kassel RL, Green S, Fiore N, Williamson B. An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3666–3670. [PMC free article] [PubMed]
  • Green S, Dobrjansky A, Chiasson MA, Carswell E, Schwartz MK, Old LJ. Corynebacterium parvum as the priming agent in the production of tumor necrosis factor in the mouse. J Natl Cancer Inst. 1977 Nov;59(5):1519–1522. [PubMed]
  • Haranaka K, Satomi N, Sakurai A, Haranaka R. Role of first stimulating agents in the production of tumor necrosis factor. Cancer Immunol Immunother. 1984;18(2):87–90. [PubMed]
  • Haranaka K, Satomi N, Sakurai A. Antitumor activity of murine tumor necrosis factor (TNF) against transplanted murine tumors and heterotransplanted human tumors in nude mice. Int J Cancer. 1984 Aug 15;34(2):263–267. [PubMed]
  • Helson L, Green S, Carswell E, Old LJ. Effect of tumour necrosis factor on cultured human melanoma cells. Nature. 1975 Dec 25;258(5537):731–732. [PubMed]
  • Haranaka K, Satomi N. Cytotoxic activity of tumor necrosis factor (TNF) on human cancer cells in vitro. Jpn J Exp Med. 1981 Jun;51(3):191–194. [PubMed]
  • Haranaka K, Satomi N, Sakurai A, Kunii O. Role of lipid A in the production of tumor necrosis factor and differences in antitumor activity between tumor necrosis factor and lipopolysaccharide. Tohoku J Exp Med. 1984 Dec;144(4):385–396. [PubMed]
  • Green S, Dobrjansky A, Carswell EA, Kassel RL, Old LJ, Fiore N, Schwartz MK. Partial purification of a serum factor that causes necrosis of tumors. Proc Natl Acad Sci U S A. 1976 Feb;73(2):381–385. [PMC free article] [PubMed]
  • Old LJ. Cancer immunology: the search for specificity--G. H. A. Clowes Memorial lecture. Cancer Res. 1981 Feb;41(2):361–375. [PubMed]
  • Old LJ. Tumor necrosis factor. Clin Bull. 1976;6(3):118–120. [PubMed]
  • Männel DN, Moore RN, Mergenhagen SE. Macrophages as a source of tumoricidal activity (tumor-necrotizing factor). Infect Immun. 1980 Nov;30(2):523–530. [PMC free article] [PubMed]
  • Matthews N. Tumour-necrosis factor from the rabbit. II. Production by monocytes. Br J Cancer. 1978 Aug;38(2):310–315. [PMC free article] [PubMed]
  • Satomi N, Haranaka K, Kunii O. Research on the production site of tumor necrosis factor (TNF). Jpn J Exp Med. 1981 Dec;51(6):317–322. [PubMed]
  • Kull FC, Jr, Cuatrecasas P. Necrosin: purification and properties of a cytotoxin derived from a murine macrophage-like cell line. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7932–7936. [PMC free article] [PubMed]
  • Pennica D, Nedwin GE, Hayflick JS, Seeburg PH, Derynck R, Palladino MA, Kohr WJ, Aggarwal BB, Goeddel DV. Human tumour necrosis factor: precursor structure, expression and homology to lymphotoxin. Nature. 1984 Dec 20;312(5996):724–729. [PubMed]
  • Shirai T, Yamaguchi H, Ito H, Todd CW, Wallace RB. Cloning and expression in Escherichia coli of the gene for human tumour necrosis factor. Nature. 313(6005):803–806. [PubMed]
  • Wang AM, Creasey AA, Ladner MB, Lin LS, Strickler J, Van Arsdell JN, Yamamoto R, Mark DF. Molecular cloning of the complementary DNA for human tumor necrosis factor. Science. 1985 Apr 12;228(4696):149–154. [PubMed]
  • Marmenout A, Fransen L, Tavernier J, Van der Heyden J, Tizard R, Kawashima E, Shaw A, Johnson MJ, Semon D, Müller R, et al. Molecular cloning and expression of human tumor necrosis factor and comparison with mouse tumor necrosis factor. Eur J Biochem. 1985 Nov 4;152(3):515–522. [PubMed]
  • Fransen L, Müller R, Marmenout A, Tavernier J, Van der Heyden J, Kawashima E, Chollet A, Tizard R, Van Heuverswyn H, Van Vliet A, et al. Molecular cloning of mouse tumour necrosis factor cDNA and its eukaryotic expression. Nucleic Acids Res. 1985 Jun 25;13(12):4417–4429. [PMC free article] [PubMed]
  • Pennica D, Hayflick JS, Bringman TS, Palladino MA, Goeddel DV. Cloning and expression in Escherichia coli of the cDNA for murine tumor necrosis factor. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6060–6064. [PMC free article] [PubMed]
  • Haranaka K, Satomi N, Sakurai A. Differences in tumour necrosis factor productive ability among rodents. Br J Cancer. 1984 Oct;50(4):471–478. [PMC free article] [PubMed]
  • Williamson BD, Carswell EA, Rubin BY, Prendergast JS, Old LJ. Human tumor necrosis factor produced by human B-cell lines: synergistic cytotoxic interaction with human interferon. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5397–5401. [PMC free article] [PubMed]
  • Ohki M, Nakamura T, Morita T, Iwanaga S. A new endotoxin sensitive factor associated with hemolymph coagulation system of horseshoe crab (Limulidae). FEBS Lett. 1980 Nov 3;120(2):217–220. [PubMed]
  • Haranaka K, Satomi N, Sakurai A, Nariuchi H. Purification and partial amino acid sequence of rabbit tumor necrosis factor. Int J Cancer. 1985 Sep 15;36(3):395–400. [PubMed]
  • Aggarwal BB, Kohr WJ, Hass PE, Moffat B, Spencer SA, Henzel WJ, Bringman TS, Nedwin GE, Goeddel DV, Harkins RN. Human tumor necrosis factor. Production, purification, and characterization. J Biol Chem. 1985 Feb 25;260(4):2345–2354. [PubMed]
  • Rubin BY, Anderson SL, Sullivan SA, Williamson BD, Carswell EA, Old LJ. High affinity binding of 125I-labeled human tumor necrosis factor (LuKII) to specific cell surface receptors. J Exp Med. 1985 Sep 1;162(3):1099–1104. [PMC free article] [PubMed]
  • Rubin BY, Anderson SL, Sullivan SA, Williamson BD, Carswell EA, Old LJ. Purification and characterization of a human tumor necrosis factor from the LuKII cell line. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6637–6641. [PMC free article] [PubMed]
  • Gray PW, Aggarwal BB, Benton CV, Bringman TS, Henzel WJ, Jarrett JA, Leung DW, Moffat B, Ng P, Svedersky LP, et al. Cloning and expression of cDNA for human lymphotoxin, a lymphokine with tumour necrosis activity. Nature. 1984 Dec 20;312(5996):721–724. [PubMed]
  • Aggarwal BB, Moffat B, Harkins RN. Human lymphotoxin. Production by a lymphoblastoid cell line, purification, and initial characterization. J Biol Chem. 1984 Jan 10;259(1):686–691. [PubMed]
  • Kull FC, Jr, Cuatrecasas P. Preliminary characterization of the tumor cell cytotoxin in tumor necrosis serum. J Immunol. 1981 Apr;126(4):1279–1283. [PubMed]
  • Green S, Dobrjansky A, Chiasson MA. Murine tumor necrosis-inducing factor: purification and effects on myelomonocytic leukemia cells. J Natl Cancer Inst. 1982 Jun;68(6):997–1003. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...