Outer membrane vesicles (OMV) production of Neisseria meningitidis serogroup B in batch process |
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Authors: | Sílvia Santos Luciana Juncioni de Arauz Júlia Baruque-Ramos Ivo Lebrun Sylvia Mendes Carneiro Sandra Alves Barreto Rocilda Perazzini Furtado Schenkman |
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Affiliation: | 1. Faculdade Adventista da Bahia (Northeast Brazil College), BR-101, km 197 – Capoeiruçu, Caixa Postal 18, Cachoeira, Bahia CEP 44300-000, Brazil;2. Instituto Adolfo Lutz (IAL), Av. Dr. Arnaldo, 355, São Paulo, SP CEP 01246-902, Brazil;3. Universidade de São Paulo (EACH), Av. Arlindo Béttio, 1000, São Paulo, SP CEP 03828-000, Brazil;4. Instituto Butantan (IB), Av. Vital Brazil, 1500, São Paulo, SP CEP 05503-900, Brazil |
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Abstract: | 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: 250-850rpm, and O(2) control at 10%, 20h. 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 162mg/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. |
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Keywords: | Neisseria meningitidis Outer membrane vesicles Vaccine Bioreactor systems Kinetics |
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