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Antibodies to synthetic peptides for the detection of survivin in tumor tissues

Abstract

Survivin, an endogenous protein, is a promising marker for the cancer diagnosis. The aim of the present work was to obtain antibodies specific to survivin and capable of detecting this protein in tumor tissues. Four peptides corresponding to fragments (1–22), (54–74), (80–88)–(153–165), and (118–144) of the survivin-2B sequence were selected and synthesized. Rabbits were immunized with the synthetic peptides. It has been shown that all peptides in a free state, without conjugation with a high-molecular-weight carrier, stimulate the production of antibodies capable of binding with recombinant survivin. Antipeptide antibodies were isolated from sera and their performance in the immunohistochemical detection of survivin in human tumor tissues was studied. It was shown that only antibodies to the (80–88)–(153–165) peptide bind to the survivin present in breast and bladder tumors. The ability of antibodies to this peptide to detect survivin in tumor tissue lysates was demonstrated by immunoblotting. The part of the sequence targeted by the antibodies against the (80–88)–(153–165) peptide was localized using truncated peptide fragments.

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Abbreviations

FIA:

Freund’s incomplete adjuvant

FCA:

Freund’s complete adjuvant

RecSurv:

recombinant survivin, Fmoc, 9-fluorenylmethoxycarbonyl

TIS:

triisopropylsilane

BSA:

bovine serum albumin

ELISA:

enzyme-linked immunosorbent assay

Ab:

antibodies

References

  1. Rudin, C.M. and Thomson, C.B., Annu. Rev. Med., 1997, vol. 48, pp. 267–281.

    Article  CAS  PubMed  Google Scholar 

  2. Johnson, M.E. and Howerth, E.W., Vet. Pathol., 2004, vol. 41, pp. 599–607.

    Article  CAS  PubMed  Google Scholar 

  3. Zhang, S.Q., Qiang, S.Y., Yang, W.B., Jianq, J.T., and Ji, Z.Z., Ai Zheng, 2004, vol. 35, pp. 40–47.

    CAS  Google Scholar 

  4. Wang, M., Wang, B., and Wang, X., Chin. Med. J., 2001, vol. 114, pp. 149–153.

    Google Scholar 

  5. Ferrandina, G., Legge, F., Martinelli, E., Raneletti, F.O., Zannoni, G.F., Lauriola, L., Gessi, M., Gallota, V., and Scambia, G., Br. J. Cancer, 2005, vol. 92, pp. 271–277.

    CAS  PubMed  Google Scholar 

  6. Tarkowski, R., Kotarski, J., Polak, G., and Wojcierowski, J., Ginecol. Pol., 2001, vol. 72, pp. 1539–1542.

    CAS  Google Scholar 

  7. Mori, A., Wada, H., Nishimura, Y., Okamoto, T., Takemoto, T., and Kakishita, E., Int. J. Hematol., 2002, vol. 75, pp. 161–165.

    Article  CAS  PubMed  Google Scholar 

  8. Yu, J., Leung, W.K., Ebert, M.P., Ng, E.K., Go, M.Y., Wang, H.B., Chung, S.C., Malfertheiner, P., and Sung, J.J., Br. J. Cancer, 2002, vol. 87, pp. 91–97.

    Article  CAS  PubMed  Google Scholar 

  9. Yang, D., Zhu, Y.Q., and Qi, J., Ai Zheng, 2004, vol. 23, pp. 306–309.

    CAS  PubMed  Google Scholar 

  10. Chiou, S.K., Jones, M.K., and Tarnawski, A.S., Med. Sci. Monit., 2003, vol. 9, pp. 125–129.

    Google Scholar 

  11. Wang, W., Luo, H., and Wang, A., J. Surg. Oncol., 2006, vol. 93, pp. 578–584.

    Article  CAS  PubMed  Google Scholar 

  12. Velculescu, V.E., Madden, S.L., Zhang, L., Lash, A.E., Yu, J., Rago, C., Lal, A., Wang, C.J., Beaudry, G.A., Ciriello, K.M., Cook, B.P., Dufault, M.R., Ferguson, A.T., Gao, Y., He, T.C., Hermeking, H., Hiraldo, S.K., Hwang, P.M., Lopez, M.A., Luderer, H.F., Mathews, B., Petroziello, J.M., Polyak, K., Zawel, L., and Kinzler, K.W., Nat. Genet., 1999, vol. 23, pp. 387–388.

    Article  CAS  PubMed  Google Scholar 

  13. Ambrosini, G., Adida, C., and Altieri, D.C., Nat. Med., 1997, vol. 3, pp. 917–921.

    Article  CAS  PubMed  Google Scholar 

  14. Mahotka, C., Wenzel, M., Springer, E., Gabbert, H.E., and Gerharz, C.D., Cancer Res., 1999, vol. 59, pp. 6097–6102.

    CAS  PubMed  Google Scholar 

  15. Byun, S., Yeo, W.G., Lee, S.E., and Lee, E., Urology, 2007, vol. 69, pp. 34–37.

    Article  PubMed  Google Scholar 

  16. Zaffaroni, N. and Daidone, M.G., Drug Resist. Updat, 2002, vol. 5, pp. 65–72.

    Article  CAS  PubMed  Google Scholar 

  17. Karam, J.A., Lotan, Y., Ashfaq, R., Sagalowsky, A.I., and Shariat, S.F., Urology, 2007, vol. 70, pp. 482–486.

    Article  PubMed  Google Scholar 

  18. Sohn, D.M., Kim, S.Y., Baek, M.J., Lim, C.W., Lee, M.H., Cho, M.S., and Kim, T.Y., Biomed. Pharmacother., 2006, vol. 60, pp. 289–292.

    Article  CAS  PubMed  Google Scholar 

  19. Rayn, B.M., Konecny, G.E., Kahlert, S., Wang, H.-J., Untch, M., Meng, G., Pegram, M.D., Podratz, K.C., Crown, J., Slamon, D.J., and Duffy, M.J., Ann. Oncol., 2006, vol. 17, pp. 597–604.

    Article  Google Scholar 

  20. Tan, H.-Y., Liu, J., Wu, S.-M., and Luo, H.-S., World J. Gastroenterol., 2005, vol. 11, pp. 4689–4692.

    CAS  PubMed  Google Scholar 

  21. Fields, A.C., Cotsonis, G., Sexton, D., Santoianni, R., and Cohen, C., Modern Pathology, 2004, vol. 17, pp. 1378–1385.

    Article  CAS  PubMed  Google Scholar 

  22. Mahotka, C., Liebmann, J., Wenzel, M., Suschek, C.V., Schmitt, M., Gabbert, H.E., and Gerharz, C.D., Cell Death Differ., 2002, vol. 9, pp. 1334–1342.

    Article  CAS  PubMed  Google Scholar 

  23. Shinohara, E.T., Gonzalez, A., Massion, P.P., Chen, H., Li, M., Freyer, A.S., Olson, S.J., Andersen, J.J., Shyr, Y., Carbone, D.P., Johnson, D.H., Hallahan, D.E., and Lu, B., Cancer, 2005, vol. 103, pp. 1685–1692.

    Article  CAS  PubMed  Google Scholar 

  24. Ponnelle, T., Chapusot, C., Martin, L., Bouvier, A.M., Plenchette, S., Faivre, J., Solary, E., and Piard, F., J. Cancer Res. Clin. Oncol., 2005, vol. 131, pp. 504–510.

    Article  CAS  PubMed  Google Scholar 

  25. Trieb, K., Lehner, R., Stulnig, T., Sulzbacher, I., and Shroyer, K., EJSO, 2003, vol. 29, pp. 379–382.

    Article  CAS  PubMed  Google Scholar 

  26. Vischioni, B., Oudejans, J.J., and Kruyt, F.A.E., Histopathology, 2007, vol. 50, pp. 659–680.

    Article  Google Scholar 

  27. Chantalat, L., Skoufias, D.A., Kleman, J.P., Jung, B., Dideberg, O., and Margolis, R.L., Mol. Cell, 2000, vol. 6, pp. 183–189.

    Article  CAS  PubMed  Google Scholar 

  28. http://antherprot-pbil.ibcp.fr

  29. Vol’pina O.M., Titova M.A., Zhmak M.N., Koroev D.O., Oboznaya M.B., Volkova, T.D. and Ivanov, V.T., Bioorgan. Khim., 2002, vol. 28, pp. 387–395. [Russ. J. Bioorg. Chem. (Engl. Transl.), 2002, vol. 28, pp. 349–356].

    Google Scholar 

  30. Deveraux, Q.L. and Reed, J.C., Genes Dev., 1999, vol. 13, pp. 239–252.

    Article  CAS  PubMed  Google Scholar 

  31. Volkova, T.D., Koroev, D.O., Titova, M.A., Oboznaya, M.V., Filatova, M.R., Pankratov, A.A., Morozova, N.V., Zolotavkina, Yu.V., Yakubovskaya, R.I., and Vol’pina, O.M., Bioorgan. Khim., 2008, vol. 34, pp. 457–463 [Russ. J. Bioorg. Chem. (Engl. Transl.), 2008, vol. 34, pp. 409–414].

    CAS  Google Scholar 

  32. Koroev, D.O., Kotelnikova, O.V., Vol’pina, O.M., Zhmak, M.N., Kuprianova, M.A., Agafonova, S.A., Alliluev, A.R., Litvinov, I.S., Nesmeynov, V.A., and Ivanov, V.T., Bioorgan. Khim., 2000, vol. 26, pp. 323–329 [Russ. J. Bioorg. Chem. (Engl. Transl.), 2000, vol. 26, pp. 292–296].

    CAS  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  34. Lowry, O.H., Rosenbrough, N.G., Farr, A.L., and Randall, R.J., J. Biol. Chem., 1951, vol. 193, pp. 265–272.

    CAS  PubMed  Google Scholar 

  35. Vladimirova, N.M., Potapenko, N.A., Sachs, G., and Modyanov, N.N., Biochim. Biophys. Acta, 1995, vol. 1233, pp. 175–184.

    Article  PubMed  Google Scholar 

  36. Zavalishina, L.E., Andreeva, Yu.Yu., and Frank, G.A., Prakticheskoe posobie (A Practical Course), Moscow, 2006.

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Correspondence to E. V. Akhidova.

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Original Russian Text © E.V. Akhidova, T.D. Volkova, D.O. Koroev, Ya.S. Kim, M.P. Filatova, N.M. Vladimirova, T.A. Karmakova, L.E. Zavalishina, Yu.Yu. Andreeva, O.M. Volpina, 2010, published in Bioorganicheskaya Khimiya, 2010, Vol. 36, No. 2, pp. 178–186.

This article is dedicated to the 50th anniversary of IBCh RAS.

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Akhidova, E.V., Volkova, T.D., Koroev, D.O. et al. Antibodies to synthetic peptides for the detection of survivin in tumor tissues. Russ J Bioorg Chem 36, 164–171 (2010). https://doi.org/10.1134/S1068162010020044

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  • DOI: https://doi.org/10.1134/S1068162010020044

Keywords

  • survivin
  • synthetic peptides
  • antibodies