title: Polysaccharide purification from Haemophilus influenzae type b through tangential microfiltration.
abstract: Haemophilus influenzae type b (Hib) is a human pathogen that causes meningitis in infants worldwide. Capsular polysaccharide linked to a protein has been used as an efficient vaccine, and this approach has reduced the incidence of Hib disease since its inclusion in national immunisation campaigns. The traditional polysaccharide downstream process is based on several ethanol precipitations, treatment with detergents and centrifugation. The aim of this study was to introduce tangential microfiltration (TMF) in the place of centrifugation to simplify handling and to scale up the process. The purity of the polysaccharide was RPNA=1747.2 and RPPrt=196.1 for nucleic acid and protein, respectively, meeting the quality requirements for this polysaccharide. Moreover, the polysaccharide was recognised by at specific antibody, and the ribose and phosphate contents were within the expected limits. Thus, we established a process for the purification of capsular polysaccharide produced by H. influenzae type b that is effective, robust and feasible to be scaling up.
authors.affiliation: Laboratório de Bioprocessos, Centro de Biotecnologia, Instituto Butantan, Brazil.
authors.forename: Silvia Maria Ferreira
authors.initials: SM
authors.lastname: Albani
authors.affiliation: Laboratório de Bioprocessos, Centro de Biotecnologia, Instituto Butantan, Brazil.
authors.forename: Mateus Ribeiro
authors.initials: MR
authors.lastname: da Silva
authors.affiliation: Laboratório Especial Piloto de Produtos Biológicos Recombinantes, Instituto Butantan, Brazil.
authors.forename: Fernando
authors.initials: F
authors.lastname: Fratelli
authors.affiliation: Laboratório Especial Piloto de Produtos Biológicos Recombinantes, Instituto Butantan, Brazil.
authors.forename: Celso Preto Cardoso
authors.initials: CP
authors.lastname: Junior
authors.affiliation: Divisão de Desenvolvimento Tecnológico e Produção, Instituto Butantan, Brazil.
authors.forename: Dmitri
authors.initials: D
authors.lastname: Iourtov
authors.affiliation: Laboratório de Bioprocessos, Centro de Biotecnologia, Instituto Butantan, Brazil.
authors.forename: Felipe de Oliveira
authors.initials: Fde O
authors.lastname: Cintra
authors.affiliation: Laboratório de Bioprocessos, Centro de Biotecnologia, Instituto Butantan, Brazil. Electronic address: mickie.takagi@butantan.gov.br.
authors.forename: Mickie
authors.initials: M
authors.lastname: Takagi
authors.affiliation: Laboratório de Bioprocessos, Centro de Biotecnologia, Instituto Butantan, Brazil.
authors.forename: Joaquin
authors.initials: J
authors.lastname: Cabrera-Crespo
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis; D019149:Bioreactors; D011232:Chemical Precipitation; D005374:Filtration; D019741:Haemophilus influenzae type b / Q000378:metabolism; D009696:Nucleic Acids / Q000032:analysis; D010758:Phosphorus / Q000032:analysis; D011135:Polysaccharides, Bacterial / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D009696:Nucleic Acids; D011135:Polysaccharides, Bacterial; D010758:Phosphorus
keywords: Downstream process; Glicoconjugate; Haemophilus influenzae type b; Polysaccharide; Tangential flow microfiltration; Vaccine