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Estimation of methods of protein delivery into mammalian cells — A comparative study by electroporation and Bioporter assay

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Abstract

To test the efficiency of some alternative methods for protein delivery into live cells were used rEGFP (recombinant Enhanced Green Fluorescent Protein) and HeLa cells as model system. The compared methods, electroporation (up to 10%) and Bioporter (up to 20%), deliver proteins and peptides to the cells with efficiency, enough for functional studies of mammalian cells. Thus, the delivery by either of both methods, followed by fluorescence activated cell sorting (FACS) of the cell population, can give a sufficient number of uniformly loaded cells for different studies including transcription assays and gene therapy.

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References

  1. Zheng, X., Lundberg, M., Karlsson, A., and Johansson, M., Cancer Res., 2003, vol. 63, no. 20, pp. 6909–6913.

    PubMed  CAS  Google Scholar 

  2. Raptis, L.H., Brownell, H.L., Liu, S.K., Firth, K.L., MacKenzie, L.W., Stiles, P.D., and Alberta, J.A., Mol. Biotechnol., 1995, vol. 4, no. 2, pp. 129–138.

    Article  PubMed  CAS  Google Scholar 

  3. Raptis, L.H., Liu, S.K., Firth, K.L., Stiles, C.D., and Alberta, J.A., BioTechniques, 1995, vol. 18, no. 1, pp. 104, 106, 108, 110 passim.

    PubMed  CAS  Google Scholar 

  4. Bright, G.R., Kuo, N.T., Chow, D., Burden, S., Dowe, C., and Przybylski, R.J., Cytometry, 1996, vol. 24, no. 3, 226–233.

    Article  PubMed  CAS  Google Scholar 

  5. Potts, R.O., Bommannan, D., Wong, O., Tamada, A., Riviere, E., and Monteiro-Riviere, A., Pharm. Biotechnol., 1997, vol. 10, pp. 213–238.

    Article  PubMed  CAS  Google Scholar 

  6. Coulberson, A.L., Hud, N.V., LeDoux, J.M., Vilfan, I.D., and Prausnitz, M.R., J. Control. Release, 2003, vol. 86, nos. 2/3, pp. 361–370.

    Article  PubMed  CAS  Google Scholar 

  7. Eksioglu-Demiralp, E., Kitada, S., Carson, D., Garland, J., Andreef, M., and Reed, J.C., J. Immunol. Methods, 2003, vol. 275, nos. 1/2, pp. 41–56.

    Article  PubMed  CAS  Google Scholar 

  8. Mashiba, H., Ozaki, Y., Ikuno, S., and Matsunaga, K., Cancer, Biother. Radiopharm., 2001, vol. 16, no. 6, pp. 495–499.

    Article  CAS  Google Scholar 

  9. Nakanishi, O., Shibasaki, F., Hidaka, M., Homma, Y., and Takenawa, T., J. Biol. Chem., 1993, vol. 268, pp. 10754–10759.

    PubMed  CAS  Google Scholar 

  10. Liao, F., Shin, H.S., and Rhee, S.G., Biochem. Biophys. Res. Commun., 1993, vol. 19, no. 3, pp. 1028–1033.

    Article  Google Scholar 

  11. Marrero, M.B., Schieffer, B., Paxton, W.G., Schieffer, E., and Bernstein, K.E., J. Biol. Chem., 1995, vol. 270, no. 26, pp. 15734–15738.

    Article  PubMed  CAS  Google Scholar 

  12. Lukas, J., Bartek, J., and Strauss, M., J. Immunol. Methods, 1994, vol. 17, no. 2, pp. 255–259.

    Article  Google Scholar 

  13. Graessmann, M. and Graessmann, A., Meth. Enzymol., 1983, vol. 101, pp. 482–492.

    Article  PubMed  CAS  Google Scholar 

  14. Handbook of Cell-Penetrating Peptides, Langel, U., Ed., 2nd ed., Boca Raton: CRC Press, Taylor Francis Group, 2007.

    Google Scholar 

  15. Watanabe, S. and Wakasugi, K., Biochem. Biophys. Res. Commun., 2008, vol. 369, no. 2, pp. 695–700.

    Article  PubMed  CAS  Google Scholar 

  16. Okada, C.Y. and Rechsteiner, M., Cell, 1982, vol. 29, no. 1, pp. 33–41.

    Article  PubMed  CAS  Google Scholar 

  17. Brown, A.D., Lopez-Terrada, D., Denny, C., and Lee, K.A., Oncogene, 1995, vol. 10, no. 9, pp. 1749–1756.

    PubMed  CAS  Google Scholar 

  18. Wadia, J.S. and Dowdy, S.F., Adv. Drug Deliv. Rev., 2005, vol. 57, pp. 579–596.

    Article  PubMed  CAS  Google Scholar 

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Correspondence to R. Todorova.

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Todorova, R. Estimation of methods of protein delivery into mammalian cells — A comparative study by electroporation and Bioporter assay. Appl Biochem Microbiol 45, 444–448 (2009). https://doi.org/10.1134/S0003683809040176

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  • DOI: https://doi.org/10.1134/S0003683809040176

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