title: Development of a whole cell pneumococcal vaccine: BPL inactivation, cGMP production, and stability.
abstract: 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 °C for 30 h. BPL was hydrolyzed by heating for 2h at 37 °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.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil.
authors.forename: Viviane M
authors.initials: VM
authors.lastname: Gonçalves
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil.
authors.forename: Waldely O
authors.initials: WO
authors.lastname: Dias
authors.affiliation: Laboratório Especial Piloto de Produtos Biológicos Recombinantes, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação Interunidades em Biotecnologia-USP-IPT-IB, São Paulo, Brazil.
authors.forename: Ivana B
authors.initials: IB
authors.lastname: Campos
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil.
authors.forename: Celia
authors.initials: C
authors.lastname: Liberman
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil.
authors.forename: Maria E
authors.initials: ME
authors.lastname: Sbrogio-Almeida
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação Interunidades em Biotecnologia-USP-IPT-IB, São Paulo, Brazil.
authors.forename: Eliane P
authors.initials: EP
authors.lastname: Silva
authors.affiliation: Laboratório Especial Piloto de Produtos Biológicos Recombinantes, Instituto Butantan, São Paulo, Brazil.
authors.forename: Celso P
authors.initials: CP
authors.lastname: Cardoso
authors.affiliation: PATH, Seattle, WA, United States.
authors.forename: Mark
authors.initials: M
authors.lastname: Alderson
authors.affiliation: PATH, Seattle, WA, United States.
authors.forename: George
authors.initials: G
authors.lastname: Robertson
authors.affiliation: PATH, Seattle, WA, United States.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Maisonneuve
authors.affiliation: PATH, Seattle, WA, United States.
authors.forename: Andrea
authors.initials: A
authors.lastname: Tate
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Boston Children's Hospital, Boston, MA 02115, United States.
authors.forename: Porter
authors.initials: P
authors.lastname: Anderson
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Boston Children's Hospital, Boston, MA 02115, United States.
authors.forename: Richard
authors.initials: R
authors.lastname: Malley
authors.affiliation: Laboratório Especial Piloto de Produtos Biológicos Recombinantes, Instituto Butantan, São Paulo, Brazil.
authors.forename: Fernando
authors.initials: F
authors.lastname: Fratelli
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil. Electronic address: luciana.leite@butantan.gov.br.
authors.forename: Luciana C C
authors.initials: LC
authors.lastname: Leite
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D019149:Bioreactors; D005260:Female; D005285:Fermentation; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D050296:Microbial Viability; D011008:Pneumococcal Infections / Q000517:prevention & control*; D022242:Pneumococcal Vaccines / Q000096:biosynthesis* / Q000276:immunology; D011420:Propiolactone / Q000494:pharmacology*; D011786:Quality Control
chemical_list: D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D022242:Pneumococcal Vaccines; D011420:Propiolactone
keywords: Streptococcus pneumoniae; Whole cell vaccine; cGMP production