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Recombinant Proteins in Vaccine Development

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Part of the Methods in Molecular Medicine™ book series (MIMM,volume 66)

Abstract

The outer membrane of Neisseria meningitidis contains a variety of proteins with the potential for inclusion in new meningococcal vaccines (1). Studies on the vaccine potential of these proteins would be facilitated by the production of pure recombinant protein, free from other components of the Neisseria outer membrane. At present, the class 1 outer-membrane protein (OMP) is generally regarded as the most promising candidate. In this chapter, we describe four protocols involved in the preparation of recombinant class 1 OMP for vaccine development. This integrated set of methods can also be readily used to study the potential of other meningococcal OMP as vaccine candidates, and moreover, their utilities make them attractive for vaccine studies relating to many other human pathogens.

Keywords

  • Recombinant Protein
  • Polymerase Chain Reaction Product
  • Luria Bertani
  • Size Exclusion Chromatography
  • Neisseria Meningitidis

These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Poolman, J. T., van der Ley, P. A., and Tommassen, J. (1995) Surface structures and secreted products of meningococci, in Meningococcal Disease (Cartwright, K. A. V., ed.), John Wiley and Sons, Chichester, UK, pp. 21–34.

    Google Scholar 

  2. Barlow, A. K., Heckels, J. E., and Clarke, I. N. (1987) Molecular cloning and expression of Neisseria meningitidis class 1 outer membrane protein in Escherichia coli K12. Infect. Immun. 55, 2734–2740.

    CAS  PubMed  Google Scholar 

  3. Barlow, A. K., Heckels, J. E., and Clarke, I. N. (1989) The class 1 outer membrane protein of Neisseria meningitidis: gene sequence and structural and immunological similarities to gonococcal porins. Mol. Microbiol. 3, 131–139.

    CAS  CrossRef  PubMed  Google Scholar 

  4. White, D. A., Barlow, A. K., Clarke, I. N., and Heckels, J. E. (1990) Stable expression of meningococcal class 1 protein in an antigenically reactive form in outer membranes of Escherichia coli. Mol. Microbiol. 4, 769–776.

    CAS  CrossRef  PubMed  Google Scholar 

  5. Nurminen, M., Butcher, S., Idanpaan-Heikkila, I., Wahlstrom, E., Muttilainen, S., Runeberg-Nyman, K., et al. (1992) The class 1 outer membrane protein of Neisseria meningitidis produced in Bacillus subtilis can give rise to protective immunity. Mol. Microbiol. 6, 2499–2506.

    CAS  CrossRef  PubMed  Google Scholar 

  6. Muttilainen, S., Idanpaan-Heikkila, I., Wahlstrom, E., Nurminen, M., Makela, P. H., and Sarvas, M. (1995) The Neisseria meningitidis outer membrane protein P1 produced in Bacillus subtilis and reconstituted into phospholipid vesicles elicits antibodies to native P1 epitopes. Microb. Pathog. 18, 423–436.

    CAS  CrossRef  PubMed  Google Scholar 

  7. Muttilainen, S., Butcher, S. J., Runeberg, K., Nurminen, M., Idanpaan-Heikkila, I., Wahlstrom, E., and Sarvas, M. (1995) Heterologous production of the P1 porin of Neisseria meningitidis in Bacillus subtilis: the effect of an N-terminal extension on the presentation of native-like epitopes. Microb. Pathog. 18, 365–371.

    CAS  CrossRef  PubMed  Google Scholar 

  8. Ward, S. J., Scopes, D. A., Christodoulides, M., Clarke, I. N., and Heckels, J. E. (1996) Expression of Neisseria meningitidis class 1 porin as a fusion protein in Escherichia coli: the influence of liposomes and adjuvants on the production of a bactericidal immune response. Microb. Pathog. 21, 499–512.

    CAS  CrossRef  PubMed  Google Scholar 

  9. QIAGEN (1997) The QIAexpressionist. A handbook for high-level expression and purification of 6xHis-tagged proteins.

    Google Scholar 

  10. Christodoulides, M., Brooks, J. L., Rattue, E., and Heckels, J. E. (1998) Immunisation with recombinant class 1 outer membrane protein from Neisseria meningitidis: influence of liposomes and adjuvants on antibody avidity, recognition of native protein and the induction of a bactericidal immune response against meningococci. Microbiology 144, 3027–3037.

    CAS  CrossRef  PubMed  Google Scholar 

  11. Gregoriadis, G. (1995) Liposomes as immunological adjuvants, in The Theory and Practical Application of Adjuvants (Stewart-Tull, D. E. S., ed.), John Wiley and Sons Ltd., Chichester, UK, pp. 145–169.

    Google Scholar 

  12. Tinsley, C. R. and Heckels, J. E. (1986) Variation in the expression of pili and outer membrane protein by Neisseria meningitidis during the course of meningococcal infection. J. Gen. Microbiol. 132, 2483–2490.

    CAS  PubMed  Google Scholar 

  13. Ausebel, F. M., Brent, R., Kingston, R. E., Moore, D. D, Sedman, J. G, Smith, J. A, and Struhl, K. (1995) Current Protocols in Molecular Biology, John Wiley and Sons, New York.

    Google Scholar 

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© 2001 Humana Press Inc., Totowa, NJ

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Christodoulides, M., Jolley, K.A., Heckels, J.E. (2001). Recombinant Proteins in Vaccine Development. In: Pollard, A.J., Maiden, M.C. (eds) Meningococcal Vaccines. Methods in Molecular Medicine™, vol 66. Humana Press. https://doi.org/10.1385/1-59259-148-5:167

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  • DOI: https://doi.org/10.1385/1-59259-148-5:167

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-801-1

  • Online ISBN: 978-1-59259-148-0

  • eBook Packages: Springer Protocols