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Improvement in the Purification Process of the Capsular Polysaccharide from Haemophilus influenzae Type b by Using Tangential Ultrafiltration and Diafiltration

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Abstract

Capsular polysaccharide produced by Haemophilus influenzae b (Hib) is the main virulent agent and used as the antigen in the vaccine formulation. In this study, an improved process of polysaccharide purification was established based on tangential flow ultrafiltration using detergents (cocamidopropyl betaine and sodium deoxycholate), two selective ethanol precipitations steps, and extensive enzymatic hydrolysis as strategy. The relative purity (RP) related to protein and nucleic acids were 122∼263 and 294∼480, respectively, and compatible with the specifications established by the World Health Organization for Hib vaccine, RP ≥ 100. These results make this process simple, cheaper, efficient, environmentally friendly, and prone to be scaled up.

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References

  1. Shapiro, E. D., & Ward, J. I. (1991). Epidemiologic Reviews, 13, 113–142.

    CAS  Google Scholar 

  2. Kelly, D. F., Moxon, E. R., & Pollard, A. J. (2004). Immunology, 113, 163–174.

    Article  CAS  Google Scholar 

  3. Gotschlich, E. C., Liu, T. Y., & Artenstein, M. S. (1969). The Journal of Experimental Medicine, 129, 1349–1365.

    Article  CAS  Google Scholar 

  4. Yavordios, D., & Cousin, M. (1983). European Patent Office. EP 0 071 515 A1.

  5. Frasch, C. E. (1990). In A. Mizrahi (Ed.), Advances in biotechnological vaccines (pp. 123–145). New York: Wiley-Liss.

    Google Scholar 

  6. Hamidi, A., & Beurret, M. (2010). European Patent Office. EP 1 664 319 B1.

  7. World Health Organization WHO. (2000). Technical Report Series, 897, 27–56.

    Google Scholar 

  8. Ward, O. P. (1991). Bioprocessing. New York: Van Nostrand Reinhold.

    Book  Google Scholar 

  9. Tanizaki, M. M., Garcia, L. R., Ramos, J. B., Leite, L. C. C., Hiss, H., Furuta, J. A., et al. (1996). Journal of Microbiological Methods, 27, 19–23.

    Article  CAS  Google Scholar 

  10. Gonçalves, V. M., Takagi, M., Carmo, T. S., Lima, R. B., Albani, S. M. F., Pinto, J. V., et al. (2007). In A. V. Mendez (Ed.), Communicating current research and educational topics in applied microbiology (pp. 450–457). Badajoz: Formatex Research Center.

    Google Scholar 

  11. Takagi, M., Lima, R. B., Albani, S. M. F., Zangirolami, T. C., Tanizaki, M. M., & Cabrera-Crespo, J. (2008). Journal of Industrial Microbiology and Biotechnology, 35, 1217–1222.

    Article  CAS  Google Scholar 

  12. Takagi, M., Cabrera-Crespo, J., Zangirolami, T. C., Raw, I., & Tanizaki, M. M. (2006). Journal of Chemical Technology and Biotechnology, 81, 182–188.

    Article  CAS  Google Scholar 

  13. Silva, M.R. (2010). Dissertation, Universidade Federal de São Carlos.

  14. Ashwell, G. (1957). Methods in Enzymology, 3, 87–90.

    Article  Google Scholar 

  15. Osborn, M. J. (1963). Biochemistry, 50, 499–506.

    CAS  Google Scholar 

  16. Petsch, D., & Anspach, F. B. (2000). Journal of Biotechnology, 76, 97–119.

    Article  CAS  Google Scholar 

  17. Magalhães, P. O., Lopes, A. M., Mazzola, P. G., Rangel-Yagui, C., Penna, T. C. V., & Pessoa, A., Jr. (2007). Journal of Pharmacy and Pharmaceutical Sciences, 10, 388–404.

    Google Scholar 

  18. Rienstra, M.S., Scattergood, E.R.M., Sitrin, R.D. (1991). United States Patent US5039610.

  19. Kogan, G., Soltés, L., Stern, R., & Gemeiner, P. (2007). Biotechnology Letters, 29, 17–25.

    Article  CAS  Google Scholar 

  20. Lindahl, U. (2000). Glycoconjugate Journal, 17, 597–605.

    Article  CAS  Google Scholar 

  21. Volpi, N. (2007). Journal of Pharmaceutical Sciences, 96(12), 3168–3180.

    Article  CAS  Google Scholar 

  22. Ozbek, S., Muller, J. F., Figgemeier, E., & Stetefeld, J. (2005). Acta Crystallographica D, 61, 477–480.

    Article  Google Scholar 

  23. Chen, Y., Xu, Q., Yang, H., & Gu, G. (2001). Process Biochemistry, 36, 773–779.

    Article  CAS  Google Scholar 

  24. Chen, Y., Chen, Y. S., & Gu, G. (2004). Chemical Engineering Journal, 99, 137–143.

    Article  CAS  Google Scholar 

  25. Pinelo, M., Jonsson, G., & Meyer, A. S. (2009). Separation and Purification Technology, 70(1), 1–11.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We gratefully acknowledge the financial support from the São Paulo Research Foundation (FAPESP 2007/50082-2) and the Butantan Foundation—Brazil. Silvia M.F. Albani received a scholarship from the Secretary of Education of São Paulo State. We also thank Mr. Lourivaldo Inácio de Souza and Ms. Inês do Amaral Maurelli for the technical assistance.

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Authors and Affiliations

  1. Instituto Butantan—Centro de Biotecnologia, Av. Vital Brasil, 1500, 05503-900, São Paulo, Brazil

    Silvia Maria Ferreira Albani, Mateus Ribeiro da Silva, Mickie Takagi & Joaquin Cabrera-Crespo

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  1. Silvia Maria Ferreira Albani
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  2. Mateus Ribeiro da Silva
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  3. Mickie Takagi
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  4. Joaquin Cabrera-Crespo
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Correspondence to Mickie Takagi.

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Albani, S.M.F., da Silva, M.R., Takagi, M. et al. Improvement in the Purification Process of the Capsular Polysaccharide from Haemophilus influenzae Type b by Using Tangential Ultrafiltration and Diafiltration. Appl Biochem Biotechnol 167, 2068–2075 (2012). https://doi.org/10.1007/s12010-012-9750-4

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  • DOI: https://doi.org/10.1007/s12010-012-9750-4

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