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Purification of capsular polysaccharide from Neisseria meningitidis serogroup C by liquid chromatography.
Pato, Tânia Pinheiro; Barbosa, Antonio de Pádua R; da Silva Junior, José Godinho.
Afiliação
  • Pato TP; Departamento de Vacinas Bacterianas, Bio-Manguinhos, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil. Tania@bio.fiocruz.br
Article em En | MEDLINE | ID: mdl-16469547
Neisseria meningitidis serogroup C capsular polysaccharide (MenCPS) is an important antigen against meningococcal infection. This paper describes a new purification methodology employing liquid chromatography that resulted in a polysaccharide showing the characteristics recommended by the World Health Organization for vaccine purposes. In this method, steps of the traditional procedure that yield low recovery and use toxic materials were modified. The present process consists in the following steps: (1) continuous flow centrifugation of the culture for removal of the cells; (2) supernatant concentration by tangential filtration (100 kDa cutoff); (3) addition of 0.5% DOC, heating to 55 degrees C during 30 min and tangential filtration (100 kDa cutoff); (4) anion exchange chromatography (Source 15Q) and (5) size exclusion chromatography (Sepharose CL-4B). The polysaccharide C fraction obtained in that way was dialyzed and freeze-dried. The structural identity of the polysaccharide was demonstrated by (1)H-NMR spectrometry.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Polissacarídeos Bacterianos / Cromatografia Líquida de Alta Pressão / Cromatografia por Troca Iônica / Neisseria meningitidis Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2006 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Polissacarídeos Bacterianos / Cromatografia Líquida de Alta Pressão / Cromatografia por Troca Iônica / Neisseria meningitidis Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2006 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda