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Methods of Preparation of Recombinant Cytokines: IV. Renaturation of Recombinant Human Interleukin-3


Renaturation of recombinant human interleukin-3 produced as inclusion bodies in the transformed cells of Escherichia coli was studied and optimized. Importance was shown of removing from the protein solution the hydrophobic cellular components causing irreversible aggregation of the protein under renaturation conditions. An effect of pH on the secondary structure of the denatured protein was revealed by CD spectroscopy. It was thereby found that at pH 8.5, which is the optimal value for renaturation, the protein has the secondary structure most close to the native one. The isolation according to the scheme proposed allows preparation of interleukin-3 in 50% yield with 99% purity and biological activity 2 × 107 U/mg.

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  1. Klyushnichenko, V.E., Tikhonov, R.V., Pechenov, S.E., Yakimov, S.A., Shingarova, L.N., Korobko, V.G., and Wulfson, A.N., Protein Expr. Purif., 1998, vol. 14, pp. 261-266.

    Google Scholar 

  2. Gianella-Borradori, A., Stem Cells (Dayt), 1994, vol. 12, pp. 241-248.

    Google Scholar 

  3. Newland, A.C., Cancer Chemother. Pharmacol., 1996, vol. 38, pp. 83-88.

    Google Scholar 

  4. Lutsenko, S.V., Gurevich, A.I., Kanevskii, V.Yu., Smirnov, V.A., Nazimov, I.V., Azhikina, T.L., Chernov, I.P., Rostapshov, V.M., Sonina, N.V., and Azhaev, A.V., Bioorg. Khim., 1991, vol. 17, pp. 1649-1654.

    Google Scholar 

  5. Gurevich, A.I., Kachalina, T.A., Miroshnikov, A.I., Sverdlov, E.D., Azhikina, T.A., Rostapshov, V.M., Esipov, R.S., Wulfson, A.N., Tikhonov, R.V., and Kostromina, T.I, Pat. RF no. 2099421, 1997, Byull. Izobret., 1997, no. 9.

  6. De Bernardes Clark, E., Hevehan, D., Szela, S., and Maachupalli-Reddy, J., Biotechnol. Prog., 1998, vol. 14, pp. 47-54.

    Google Scholar 

  7. Feng, Y., Klein, B.K., and McWherter, C.A., J. Mol. Biol., 1996, vol. 259, pp. 524-541.

    Google Scholar 

  8. Wulfson, A.N., Tikhonov, R.V., Pechenov, S.E., Klyushnichenko, V.E., and Miroshnikov, A.I., Bioorg. Khim., 1997, vol. 23, pp. 721-726.

    Google Scholar 

  9. Ellman, G.L., Arch. Biochem. Biophys., 1959, vol. 82, pp. 70-77.

    Google Scholar 

  10. Freeman, J.J., Parr, G.R., Hecht, R.I., Morris, J.C., and McKearn, J.P., Int. J. Biochem., 1991, vol. 23, pp. 353-360.

    Google Scholar 

  11. Laemmli, U.K., Nature, 1970, vol. 227, pp. 680-685.

    Google Scholar 

  12. Andrade, M.A., Chacon, P., Merelo, J.J., and Moran, F., Protein Eng., 1993, vol. 6, pp. 383-390.

    Google Scholar 

  13. Merelo, J.J., Andrade, M.A., Prieto, A., and Moran, F., Neurocomputing, 1994, vol. 6, pp. 1-12.

    Google Scholar 

  14. Bradford, M.M., Anal. Biochem., 1976, vol. 72, pp. 248-254.

    Google Scholar 

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Tikhonov, R.V., Pechenov, S.E., Gurevich, A.I. et al. Methods of Preparation of Recombinant Cytokines: IV. Renaturation of Recombinant Human Interleukin-3. Russian Journal of Bioorganic Chemistry 27, 34–38 (2001).

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  • CD spectroscopy
  • HPLC
  • hydrophobic cellular components
  • inclusion bodies
  • recombinant human interleukin-3
  • renaturation
  • thiol–disulfide exchange