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Preformulation study of the vaccine candidate P64k against Neisseria meningitidis
Author(s) -
Expósito Raya Néstor,
Luaces Marissa Mestre,
Rodriguez Ricardo Silva,
Nazábal Gálvez Consuelo,
Peña Rivero Maxlenin,
Martínez de la Puente Nieves,
Batista Milagros Font,
Guillén Nieto Gerardo
Publication year - 1999
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1999.tb00540.x
Subject(s) - immunogenicity , neisseria meningitidis , microbiology and biotechnology , escherichia coli , neisseria , biology , recombinant dna , bacterial outer membrane , neisseriaceae , chemistry , gene , antigen , bacteria , genetics , antibiotics
We have previously isolated, cloned and expressed in Escherichia coli the lpdA gene coding for a high‐molecular‐mass protein (P64k) common to many meningococcal strains. P64k is an outer membrane lipoamide dehydrogenase that is highly immunogenic in animals. Here we describe a preformulation study of the recombinant protein as a vaccine candidate against Neisseria meningitidis , in which six variants containing the candidate were tested. Three assays were used to identify the most suitable variant for further evaluation: percentage of adsorption, identification of P64k by SDS/PAGE, and immunogenicity in mice. All the preformulation variants studied showed more than 98% of adsorption of P64k on the aluminium gel. After desorption, P64k was also identified by SDS/PAGE in the six preformulation variants. Seroconversion was attained in all groups analysed. On the basis of these results, the most effective variant consisted of 20 μg/ml P64k plus 0.5 mg/ml aluminium hydroxide.