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1.
Vaccine ; 30(42): 6064-9, 2012 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-22867717

RESUMO

Serogroup B outer membrane vesicles (OMV) with iron regulated proteins (IRP) from Neisseria meningitidis constitute the antigen for the vaccine against the disease caused by this bacterium. Aiming to enhance final OMV concentration, seven batch experiments were carried out under four different conditions: (i) with original Catlin medium; (ii) with original Catlin medium and lactate and amino acids pulse at the 6th cultivation hour; (iii) with Catlin medium with double initial concentrations of lactate and amino acids and (iv) Catlin medium without glycerol and with double initial concentrations of lactate and amino acids. The cultivation experiments were carried out in a 7-L bioreactor under the following conditions: 36°C, 0.5atm, overlay air 1L/min, agitation: 250-850 rpm, and O(2) control at 10%, 20 h. After lactate and amino acids exhaustion, cell growth reached stationary phase and a significant release increase of OMV was observed. According to the Luedeking & Piret model, OMV liberation is non-growth associated. Glycerol was not consumed during cultivation. The maximum OMV concentration value attained was 162 mg/L with correspondent productivity of 8.1mg/(Lh) employing Catlin medium with double initial concentrations of lactate and amino acids. The obtained OMV satisfied constitution and protein pattern criteria and were suitable for vaccine production.


Assuntos
Proteínas da Membrana Bacteriana Externa/biossíntese , Reatores Biológicos , Neisseria meningitidis Sorogrupo B/metabolismo , Vesículas Secretórias/química , Aminoácidos/química , Técnicas de Cultura Celular por Lotes , Meios de Cultura/química , Glicerol/metabolismo , Proteínas Reguladoras de Ferro/química , Ácido Láctico/química , Vacinas Meningocócicas/biossíntese
2.
J Med Microbiol ; 57(Pt 3): 338-342, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18287297

RESUMO

Identification of Neisseria meningitidis PorA types remains important, as the PorA protein is a major immunogenic component of several meningococcal vaccines under development. In this study, 191 N. meningitidis serogroup B isolates collected in Argentina through active laboratory-based surveillance from 2001 to 2003 were serosubtyped. Nucleotide sequences of the porA variable region 1 (VR1) and VR2 regions were determined in 52 non-serosubtypeable isolates. A substantial number of distinct VR types were identified, and a new VR2 variant from the P1.16 family was described. This is the first report describing PorA types in N. meningitidis serogroup B isolates in Argentina. Furthermore, the wide diversity of subtypes detected by serosubtyping and genosubtyping reveals the difficulty in designing a useful outer-membrane vaccine applicable in this country. A possible mechanism responsible for altered PorA expression was analysed in two PorA types.


Assuntos
Desenho de Fármacos , Variação Genética , Vacinas Meningocócicas , Neisseria meningitidis Sorogrupo B/classificação , Porinas/classificação , Argentina/epidemiologia , Sequência de Bases , Genótipo , Humanos , Infecções Meningocócicas/epidemiologia , Infecções Meningocócicas/microbiologia , Vacinas Meningocócicas/química , Dados de Sequência Molecular , Neisseria meningitidis Sorogrupo B/genética , Neisseria meningitidis Sorogrupo B/isolamento & purificação , Neisseria meningitidis Sorogrupo B/metabolismo , Vigilância da População , Porinas/química , Porinas/genética , Porinas/metabolismo , Regiões Promotoras Genéticas , Análise de Sequência de DNA , Sorotipagem
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