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The synthesis of TNF-alpha conjugates with alendronic acid


Conjugates of recombinant human tumor necrosis factor alpha (TNFα) and alendronic acid linked through the protein sulfhydryl, carboxyl, and amino groups were obtained with crosslinking agents of different types. The conjugation reactions were conducted in solution and on a solid phase. Unlike the conjugation reactions in solution, the method involving immobilization of active components on a hydroxyapatite column was shown to result in the conjugates with a specified stoichiometry and a high degree of homogeneity. The TNFα conjugates retained the specific cytolytic activity and demonstrated the higher affinity to hydroxyapatite, an analogue of the bone mineral matrix, than TNFα.

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alendronic acid




glutaric aldehyde




sulfosuccinimidyl-4-(Nmaleimidomethyl) cyclohexane-1-carboxylate


tumor necrosis factor


  1. Coleman, R.E., Semin. Oncol., 2007, vol. 34, no. Suppl. 4, pp. 11–S16.

    Article  Google Scholar 

  2. Ketlinskii, S.A. and Simbirtsev, A.S., Tsitokiny (Cytokines), Moscow: Foliant, 2008.

    Google Scholar 

  3. Shmelev, V.A., Interferon-gamma, faktor nekroza opukholei, timozin-al’fa1–protivoinfektsionnye i protivoopukholevye tsitokiny i preparaty (Interferon gamma, Tumor Necrosis Factor, Thymosin alpha-1—Antiinfective and Anticancer Drugs and Cytokines), Moscow: Medpraktika, 2008.

    Google Scholar 

  4. Nasonov, E.L., Skripnikova, I.A., and Nasonova, V.A., Problema osteoporoza v revmatologii (Problem of Osteoporosis in Rheumatology), Moscow: Stin, 1997.

    Google Scholar 

  5. Ishizuka, S., Kurihara, N., Hiruma, Y., Miura, D., Namekawa, J., Tamura, A., Kato-Nakamura, Y., Nakano, Y., Takenouchi, K., Hashimoto, Y., Nagasawa, K., and Roodman, G.D., J. Steroid Biochem. Mol. Biol., 2008, vol. 110, nos. 3–5, pp. 269–277.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. Frith, J.C., Mönkkönen, J., Blackburn, G.M., Russell, R.G., and Rogers, M.J., J. Bone Miner. Res., 1997, vol. 12, pp. 1358–1367.

    CAS  Article  PubMed  Google Scholar 

  7. Danilenko, E.D. and Belkina, A.O, Biofarm. Zh., 2014, vol. 6, no. 6, pp. 44–53.

    Google Scholar 

  8. Knorre, D.G. and Myzina, S.D., Biologicheskaya khimiya: Ucheb. dlya khim., biol. i med. spets. vuzov (Biological Chemistry: Textbook for Chemical, Biological, and Medical Specialities of Universities), Moscow: Vyssh. shkola, 2000.

    Google Scholar 

  9. Uludag, H., Kousinioris, N., Gao, T., and Kantoci, D, Biotechnol. Prog., 2000, vol. 16, pp. 258–267.

    CAS  Article  PubMed  Google Scholar 

  10. Uludag, H. and Yang, J, Biotechnol. Prog., 2002, vol. 18, no. 3, pp. 604–611.

    CAS  Article  PubMed  Google Scholar 

  11. Ehrick, R.S., Capaccio, M., Puleo, D.A., and Bachas, L.G, Bioconjug. Chem., 2008, vol. 19, pp. 315–321.

    CAS  Article  PubMed  Google Scholar 

  12. Pustoshilova, N.M., Kileva, E.V., Denisova, L.Ya., Shingareva, N.V., Korobko, V.G., Denisov, L.A., Masycheva, V.I., Sandakhchiev, L.S., and Kalinin, Yu.T., RF Patent No. 2144958, 1997.

    Google Scholar 

  13. Lebedev, L.R., Zernov, Yu.P., Andreeva, I.S., and Pustoshilova, N.M, Biotekhnologiya, 1995, no. 12, pp. 19–24.

    Google Scholar 

  14. Scopes, R., Metody ochistki belkov (Protein Purification Methods), Moscow: Mir, 1985.

    Google Scholar 

  15. Solov’eva, G.A., et al., Praktikum po biokhimii (A Practical Course in Biochemistry), Severin, S.E. and Solov’eva, G.A., Eds., Moscow: Izd. Mosk. Univ., 1989.

  16. Laemmli, U.K, Nature, 1970, vol. 227, pp. 680–685.

    CAS  Article  PubMed  Google Scholar 

  17. Meager, A., Leung, H., and Woolley, J., J. Immunol. Methods, 1989, vol. 116, pp. 1–17.

    CAS  Article  PubMed  Google Scholar 

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Correspondence to E. A. Volosnikova.

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Original Russian Text © E.A. Volosnikova, I.F. Demin, G.M. Levagina, L.R. Lebedev, A.I. Zakabunin, E.D. Danilenko, 2016, published in Bioorganicheskaya Khimiya, 2016, Vol. 42, No. 6, pp. 704–711.

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Volosnikova, E.A., Demin, I.F., Levagina, G.M. et al. The synthesis of TNF-alpha conjugates with alendronic acid. Russ J Bioorg Chem 42, 638–645 (2016).

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  • alendronic acid
  • binding to hydroxyapatite
  • conjugation
  • tumor necrosis factor alpha recombinant human