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.
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References
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. 21ā34.
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, 2734ā2740.
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, 131ā139.
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, 769ā776.
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, 2499ā2506.
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, 423ā436.
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, 365ā371.
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, 499ā512.
QIAGEN (1997) The QIAexpressionist. A handbook for high-level expression and purification of 6xHis-tagged proteins.
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, 3027ā3037.
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. 145ā169.
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, 2483ā2490.
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.
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Ā© 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 Medicineā¢, 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
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