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Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins
Published on: July 1, 2011
The purification and protective capacity of Bordetella pertussis outer membrane proteins
H J Hamstra1, B Kuipers, D Schijf-Evers
1National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.
Abstract:
The whole cell vaccine (WCV) of Bordetella pertussis is protective in the intracerebral (i.c.) mouse protection assay. We found a correlation between the i.c. mouse protection assay potency and the presence of the virulence-associated outer membrane proteins (OMPs) in outer membrane complexes (OMC). The virulence-associated 92, 32 and 30 kDa OMPs were purified and the N-terminal amino acid sequences were determined. The N-terminal amino acid sequences of the 30 and 32 kDa OMPs show homology with the C-terminal fragment of the P.93 precursor of the 69 kDa OMP (pertactin). The purified 32 kDa OMP was protective in the i.c. test when presented as mixed protein-detergent micelles. The 92 kDa OMP became a protective antigen when nonprotective levels of pertussis toxin were added. We found a correlation between the i.c. mouse protection value and the 92 kDal38 kDa (porin) ratio in OMC preparations.
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