Development of a whole cell pneumococcal vaccine: BPL inactivation, cGMP production, and stability

Autor(es):Mostrar menos -

Goncalves, Viviane M. ^[1] ; Dias, Waldely O. ^[1] ; Campos, Ivana B. ^{[2, 3]} ; Liberman, Celia ^[1] ; Sbrogio-Almeida, Maria E. ^[1] ; Silva, Eliane P. ^{[1, 2]} ; Cardoso, Jr., Celso P. ^[3] ; Alderson, Mark ^[4] ; Robertson, George ^[4] ; Maisonneuve, Jean-Francois ^[4] ; Tate, Andrea ^[4] ; Anderson, Porter ^[5] ; Malley, Richard ^[5] ; Fratelli, Fernando ^[3] ; Leite, Luciana C. C. ^[1]

Afiliação do(s) autor(es)

^[1] Inst Butantan, Ctr Biotecnol, Sao Paulo - Brazil

^[2] Programa Posgrad Interunidades Biotecnol USP IPT, Sao Paulo - Brazil

^[3] Inst Butantan, Lab Especial Piloto Prod Biol Recombinantes, Sao Paulo - Brazil

^[4] PATH, Seattle, WA - USA

^[5] Boston Childrens Hosp, Dept Med, Div Infect Dis, Boston, MA 02115 - USA

• Tipo de documento: Artigo Científico
• Fonte: Vaccine; v. 32, n. 9, p. 1113-1120, FEB 19 2014.
• Citações Web of Science: 15

Resumo

Pneumococcal infections impose a large burden of disease on the human population, mainly in developing countries, and the current pneumococcal vaccines offer serotype-specific protection, but do not cover all pathogenic strains, leaving populations vulnerable to disease caused by non-vaccine serotypes. The pneumococcal whole cell vaccine is a low-cost strategy based on non-capsular antigens common to all strains, inducing serotype-independent immunity. Therefore, we developed the process for the cGMP production of this cellular vaccine. Initially, three engineering runs and two cGMP runs were performed in 60-L bioreactors, demonstrating the consistency of the production process, as evaluated by the growth curves, glucose consumption and metabolite formation (lactate and acetate). Cell recovery by tangential filtration was 92 +/- 13%. We optimized the conditions for beta-propiolactone (BPL) inactivation of the bacterial suspensions, establishing a maximum cell density of OD600 between 27 and 30, with a BPL concentration of 1:4000 (v/v) at 150 rpm and 4 degrees C for 30 h. BPL was hydrolyzed by heating for 2 h at 37 degrees C. The criteria and methods for quality control were defined using the engineering runs and the cGMP Lots passed all specifications. cGMP vaccine Lots displayed high potency, inducing between 80 and 90% survival in immunized mice when challenged with virulent pneumococci. Sera from mice immunized with the cGMP Lots recognized several pneumococcal proteins in the extract of encapsulated strains by Western blot. The cGMP whole cell antigen bulk and whole cell vaccine product lots were shown to be stable for up to 12 and 18 months, respectively, based upon survival assays following i.p. challenge. Our results show the consistency and stability of the cGMP whole cell pneumococcal vaccine lots and demonstrate the feasibility of production in a developing country setting. (C) 2013 Elsevier Ltd. All rights reserved. (AU)

• Processo FAPESP: 03/07447-9 - Desenvolvimento de vacina celular anti-pneumococica: utilizacao de cepa nao capsulada de streptococcus pneumoniae.
• Beneficiário: Waldely de Oliveira Dias
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 08/10364-1 - Impacto das estratégias de cultivo de Streptococcus pneumoniae sobre a produção do polissacarídeo capsular
• Beneficiário: Viviane Maimoni Gonçalves
• Modalidade de apoio: Auxílio à Pesquisa - Regular