Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1981 Sep;33(3):763–768. doi: 10.1128/iai.33.3.763-768.1981

Radioimmunoassay for human beta interferon.

M Inoue, Y H Tan
PMCID: PMC350775  PMID: 6169643

Abstract

Pure human beta interferon (Hu IFN-beta) (formerly human fibroblast interferon) was used to raise antibodies in rabbits. Anti-IFN-beta immunoglobulins from rabbit serum were purified by IFN-beta affinity chromatography and were used in the development of a sandwich-type radioimmunoassay for Hu IFN-beta. This radioimmunoassay consisted of (i) incubating Hu IFN-beta with anti-IFN-beta immunoglobulins immobilized on Sepharose 4B beads and (ii) incubating the resulting solid-phase IFN antibody complex with 125I-labeled anti-Hu IFN-beta immunoglobulins. The amount of 125I-labeled anti-Hu IFN-beta bound to the complex was linearly proportional to the amount of antiviral activity present in the complex from 0 to 500 U of standard IFN-beta per ml; 1 U of antiviral activity was equivalent to about 40 cpm of 125I-labeled anti-IFN immunoglobulins in the radioimmunoassay. The results of the radioimmunoassay were not influenced by the protein content of the IFN sample assayed, and the degree of intraassay variability was low; coefficients of variability were 9.4% at 5 U/ml and 1.1% at 500 U/ml. Thus, compared with the traditional antiviral assay for IFN, this radioimmunoassay was equally sensitive and more precise. In addition, it was highly specific for Hu IFN-beta and took only 5 h to complete.

Full text

PDF
763

Images in this article

Selected References

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

  1. Addison G. M., Hales C. N. Two site assay of human growth hormone. Horm Metab Res. 1971 Jan;3(1):59–60. doi: 10.1055/s-0028-1096761. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Berthold W., Tan C., Tan Y. H. Purification and in vitro labeling of interferon from a human fibroblastoid cell line. J Biol Chem. 1978 Jul 25;253(14):5206–5212. [PubMed] [Google Scholar]
  4. Billiau A., Joniau M., De Somer P. Mass production of human interferon in diploid cells stimulated by poly-I:C. J Gen Virol. 1973 Apr;19(1):1–8. doi: 10.1099/0022-1317-19-1-1. [DOI] [PubMed] [Google Scholar]
  5. Ceska M., Lundkvist U. A new and simple radioimmunoassay method for the determination of IgE. Immunochemistry. 1972 Oct;9(10):1021–1030. doi: 10.1016/0019-2791(72)90112-7. [DOI] [PubMed] [Google Scholar]
  6. Crumpton M. J., Parkhouse R. M.E. Comparison of the effects of various detergents on antigen-antibody interaction. FEBS Lett. 1972 May 1;22(2):210–212. doi: 10.1016/0014-5793(72)80047-4. [DOI] [PubMed] [Google Scholar]
  7. Dimitriadis G. J. Effect of detergents on antibody-antigen interaction. Anal Biochem. 1979 Oct 1;98(2):445–451. doi: 10.1016/0003-2697(79)90165-9. [DOI] [PubMed] [Google Scholar]
  8. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gonenne A., Ernst R. Solubilization of membrane proteins by sulfobetaines, novel zwitterionic surfactants. Anal Biochem. 1978 Jun 15;87(1):28–38. doi: 10.1016/0003-2697(78)90565-1. [DOI] [PubMed] [Google Scholar]
  10. Ho M., Tan Y. H., Armstrong J. A. Accentuation of production of human interferon by metabolic inhibitors. Proc Soc Exp Biol Med. 1972 Jan;139(1):259–262. doi: 10.3181/00379727-139-36122. [DOI] [PubMed] [Google Scholar]
  11. Knight E., Jr, Hunkapiller M. W., Korant B. D., Hardy R. W., Hood L. E. Human fibroblast interferon: amino acid analysis and amino terminal amino acid sequence. Science. 1980 Feb 1;207(4430):525–526. doi: 10.1126/science.7352259. [DOI] [PubMed] [Google Scholar]
  12. Knight E., Jr Interferon: purification and initial characterization from human diploid cells. Proc Natl Acad Sci U S A. 1976 Feb;73(2):520–523. doi: 10.1073/pnas.73.2.520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knight E. Preparation of 125iodine-labelled human fibroblast interferon. J Gen Virol. 1978 Sep;40(3):681–684. doi: 10.1099/0022-1317-40-3-681. [DOI] [PubMed] [Google Scholar]
  14. Okamura H., Berthold W., Hood L., Hunkapiller M., Inoue M., Smith-Johannsen H., Tan Y. H. Human fibroblastoid interferon: immunosorbent column chromatography and N-terminal amino acid sequence. Biochemistry. 1980 Aug 5;19(16):3831–3835. doi: 10.1021/bi00557a028. [DOI] [PubMed] [Google Scholar]
  15. Readhead C., Addison G. M., Hales C. N., Lehmann H. Immunoradiometric and two-site assay of human follicle-stimulating hormone. J Endocrinol. 1973 Nov;59(2):313–323. doi: 10.1677/joe.0.0590313. [DOI] [PubMed] [Google Scholar]
  16. Tan Y. H., Armstrong J. A., Ke Y. H., Ho M. Regulation of cellular interferon production: enhancement by antimetabolites. Proc Natl Acad Sci U S A. 1970 Sep;67(1):464–471. doi: 10.1073/pnas.67.1.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. YALOW R. S., BERSON S. A. Immunoassay of endogenous plasma insulin in man. J Clin Invest. 1960 Jul;39:1157–1175. doi: 10.1172/JCI104130. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES