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ANANBIOANAL - 2010
Pharmaceutical R & D Summit
doi:10.4172/2155-9872.1000065
In Deep Proteomic Characterization of an OMV Based Vaccine
Against
Jeovanis Gil Valdés
Department of Proteomics, Biomedical Researches, Centre for Genetic Engineering and Biotechnology,
Havana, Cuba
A
t present one of the efforts devoted to the development of an effective vaccine
against Neisseria meningitidis serogroup B have been focused in the proteomic
studies of outer membrane vesicles (OMV). This work presents the results from a
detailed study of the protein composition of OMV from VA-MENGOC-BC™
(Finlay
Institute, Cuba). The characterization of this protein preparation is a challenge
because of it is enriched in lipids and membrane proteins and only a few number
of proteins represent about 70 % of the total protein mass. Proteins were identified
by, combining two-dimensional gel electrophoresis of proteins and peptides and
mass spectrometry, and the application of non-gel based approaches starting form
the tryptic digest of the OMV preparation followed by (1) the Selective Capture
of Peptides (SCAPE-nHnR), and (2) equalization of the peptide mixture by using
peptide libraries and fractionation by reverse phase liquid chromatography at basic
pH before LC-MS/MS analysis. Most of the applied methods have been developed
in our group. This study resulted in the identification of more than three hundred
proteins. Bioinformatics analysis of the identified components allowed the selection
of potential antigens for cloning, expression, purification and immunological studies,
which were part of a wider project aimed to the development of a new vaccine
based on a defined protein composition.
ANALBIOANAL-2010
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