1887

Abstract

Human placental trophoblast interferon (tro-IFN), induced in trophoblast cultures by a superinduction procedure, was purified to a homogeneous product with retention of biological activity. The problems associated with isolation from serum-containing medium were overcome by a combination of Blue Sepharose affinity chromatography and reversed-phase HPLC (RP-HPLC) on Separon SGX C-18. This two-step purification procedure yielded tro-IFN with a specific activity of 3·4 × 10 international units/mg of protein. The overall recovery of interferon activity was 66·7%. The purified tro-IFN was shown to be a glycoprotein with an of 24K on native and SDS-PAGE. Its antiviral activity was stable at pH 2·0 at 37 °C but was sensitive to heat at 56 °C for 1 h and was neutralized by antibodies to human IFN-.

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1990-12-01
2024-11-13
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References

  1. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  2. Cesario T. C., Schryer P. J., Tilles J. G. 1977; Relationship between the physicochemical nature of human interferon, the cell induced, and the inducing agent. Antimicrobial Agents and Chemotherapy 11:291–298
    [Google Scholar]
  3. Davey M. W., Sulkowski E., Carter W. A. 1976; Binding of human fibroblast interferon to concanavalin A-agarose. Involvement of carbohydrate recognition and hydrophobic interaction. Biochemistry 15:704–713
    [Google Scholar]
  4. Doerner K. C., White B. A. 1990; Detection of glycoproteins separated by nondenaturating polyacrylamide gel electrophoresis using the periodic acid-Schiff stain. Analytical Biochemistry 187:147–150
    [Google Scholar]
  5. Duc-Goiran P., Robert-Galliot B., Chudzio T., Chany C. 1983; Unusual human interferons produced by virus-infected amniotic membranes. Proceedings of the National Academy of Sciences, U.S.A 80:2628–2631
    [Google Scholar]
  6. Fantes K. H. 1973; Purification and physico-chemical properties of interferons. In Interferons and Interferon Inducers pp. 171–200 Finter N. B. Edited by New York: North-Holland;
    [Google Scholar]
  7. Fisher P. B., Grant S. 1985; Effects of interferon on differentiation of normal and tumor cells. Pharmacology and Therapy 27:143–166
    [Google Scholar]
  8. Heine J. W., Van Damme J., De Ley M., Billiau A., De Somer P. 1981; Purification of human fibroblast interferon by zinc chelate chromatography. Journal of General Virology 54:47–56
    [Google Scholar]
  9. Kirchner H. 1986; The interferon system as an integral part of the defence system against infections. Antiviral Research 6:1–17
    [Google Scholar]
  10. Klein J. O., Remington J. S., Marcy S. M. 1976; An introduction to infections of the foetus and newborn infant. In Infectious Diseases of the Foetus and Newborn Infants pp. 1–32 Remington J. S., Klein J. O. Edited by Philadelphia: W. B. Saunders;
    [Google Scholar]
  11. Kliman H., Feinman M. A., Strauss J. F. 1987; Differentiation of human cytotrophoblast into syncytrophoblast in culture. Trophoblast Research 2:407–421
    [Google Scholar]
  12. Knight E. Jr 1976; Interferon: purification and initial characteri-zation from diploid cells. Proceedings of the National Academy of Sciences, U.S.A 73:520–523
    [Google Scholar]
  13. Knight E. Jr Fahey D. 1980; Human fibroblast interferon: an improved purification. Journal of Biological Chemistry 256:3609–3611
    [Google Scholar]
  14. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  15. Marshall L. W., Pitha P. M., Carter W. A. 1972; Inactivation of interferon: the effect of protonation. Virology 48:607–611
    [Google Scholar]
  16. Ng M. H., Vilček J. 1972; Interferons: physicochemical properties and control of cellular synthesis. Advances in Protein Chemistry173–234
    [Google Scholar]
  17. Okamura H., Berthold W., Hood L., Hunkapiller M., Inove M., Smith-Johannsen H., Tan Y. H. 1980; Human fibroblastoid interferon: immunosorbent column chromatography and N-terminal amino acid sequence. Biochemistry 19:3831–3835
    [Google Scholar]
  18. Otto M. J., Sedmak J. J., Grossberg S. E. 1980; Enzymatic modifications of human fibroblast and leukocyte interferons. Journal of Virology 35:390–399
    [Google Scholar]
  19. Remington J. S., Desmonts G. 1976; Toxoplasmosis. In Infectious Diseases of the Foetus and Newborn Infants pp. 191–332 Remington J. S., Klein J. O. Edited by Philadelphia: W. B. Saunders;
    [Google Scholar]
  20. Reynolds F. H. Jr Pitha P. M. 1975; Molecular weight study of human fibroblast interferon. Biochemical and Biophysical Research Communications 65:107–111
    [Google Scholar]
  21. Rubinstein M., Rubinstein S., Familleti P. C., Gross M. S., Miller R. S., Waldman A. A., Pestka S. 1978; Human leukocyte interferon purified to homogeneity. Science 202:1289–1290
    [Google Scholar]
  22. Sedmak J. J., Grossberg S. E. 1977; Stabilization of interferons. In Texas Reports on Biology and Medicine 35 pp. 198–203 Baron S., Dianzani F. Edited by Galveston: University of Texas Medical Branch;
    [Google Scholar]
  23. Tan Y. H., Berthold W. 1977; A mechanism for the induction and regulation of human fibroblastoid interferon genetic expression. Journal of General Virology 34:401–411
    [Google Scholar]
  24. Tan Y. H., Barakat F., Berthold W., Smith-Johannsen H., Tan C. 1979; The isolation and amino acid/sugar composition of human fibroblastoid interferon. Journal of Biological Chemistry 254:8067–8073
    [Google Scholar]
  25. Tóth F. D., Juhl C., Nørskov-Lauritsen N., Petersen P. M., Ebbesen P. 1990a; Interferon production by cultured human trophoblast induced with double stranded polyribonucleotide. Journal of Reproductive Immunology 17:217–227
    [Google Scholar]
  26. Tóth F. D., Juhl C., Nørskov-Lauritsen N., Petersen P. M., Ebbesen P. 1990b; Human trophoblast interferon: pattern of response to priming and superinduction of purified term trophob- lasts and choriocarcinoma cells. Journal of Reproductive Immunology in press
    [Google Scholar]
  27. Yamauchi T., Wilson C., St Geme J. W. Jr 1974; Transmission of live, attenuated mumps virus to the human placenta. New England Journal of Medicine 290:710–712
    [Google Scholar]
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