Rapid characterization of outer-membrane proteins in Neisseria lactamica by SELDI-TOF-MS (surface-enhanced laser desorption ionization-time-of-flight MS) for use in a meningococcal vaccine.
Immunological and epidemiological evidence suggests that the development of natural immunity to meningococcal disease results from colonization of the nasopharynx by commensal Neisseria species, particularly with Neisseria lactamica. We have reported previously that immunization with N. lactamica outer-membrane vesicles containing the major OMPs (outer-membrane proteins) protected mice against lethal challenge with meningococci of diverse serogroups and serotypes and has the potential to form the basis of a vaccine against meningococcal diseases [Oliver, Reddin, Bracegirdle et al. (2002) Infect. Immun. 70, 3621-3626]. In the present study, we have shown that biomass production and the profile of outer-membrane vesicle proteins may be affected by fermentation conditions and, in particular, media composition. Ciphergen SELDI-TOF Protein Chips were used as a rapid and sensitive new method in comparison with conventional SDS/PAGE. SELDI-TOF-MS (surface-enhanced laser-desorption ionization-time-of-flight MS) reproducibly identified three major OMPs (NspA, RmpM and PorB) and detected the changes in the protein profile when the growth medium was altered. The findings of this work indicate that SELDI-TOF-MS is a useful tool for the rapid optimization of OMP production in industrial fermentation processes and can be adapted as a Process Analytical Technology.
Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, U.K.
Tarit Kumar
TK
Mukhopadhyay
Denise
D
Halliwell
Cliona
C
O'Dwyer
Parviz Ayazi
PA
Shamlou
Myriam Susana
MS
Levy
Nigel
N
Allison
Andrew
A
Gorringe
Karen M
KM
Reddin
D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D022401:Meningococcal Vaccines / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D042806:Neisseria lactamica / Q000737:chemistry / Q000276:immunology*; D009345:Neisseria meningitidis / Q000737:chemistry / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D022401:Meningococcal Vaccines; C105506:NspA protein, Neisseria