DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14

Autor(es)

R. T. Zanardo ; A. L. S. Ferri ; D. B. Figueiredo ; S. Kraschowetz ; J. Cabrera-Crespo ^[5] ; V. M. Gonçalves ^[6]

• Tipo de documento: Artigo Científico
• Fonte: Brazilian Journal of Chemical Engineering; v. 33, n. 3, p. 435-443, 2016-09-00.

Resumo

Abstract The main virulence factor of Streptococcus pneumoniae is the capsular polysaccharide (PS), which is the antigen of all current vaccines that are prepared with PS purified from serotypes prevalent in the population. In this work, three purification strategies were evaluated and a new process was developed for purification of serotype 14 PS (PS14), responsible for 39.8% of diseases in children of 0-6 years old in Brazil. The developed method consists of cell separation by tangential microfiltration, concentration of the microfiltrate by tangential ultrafiltration (50 kDa), diafiltration in the presence of sodium dodecyl sulfate using a 30 kDa ultrafiltration membrane, precipitation with 5% trichloroacetic acid, precipitation with 20% and 60% ethanol, and anion exchange chromatography. The required purity regarding nucleic acids (≤ 2%) and proteins (≤ 3%) was achieved, resulting in a relative purity of 439 mg PS14/mg nucleic acids and 146 mg PS14/mg proteins. The final polysaccharide recovery was 65%, which is higher than the recovery of the majority of processes described in the literature. (AU)

• Processo FAPESP: 08/05207-4 - Vacina conjugada anti-pneumocócica: estudos sobre a viabilidade de uma vacina polissacarídeo capsular - PSPA
• Beneficiário: Martha Massako Tanizaki
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• 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
• Processo FAPESP: 12/04858-7 - Desenvolvimento do processo de purificação da proteína a de superfície do pneumococo do clado 4 (PspA4Pro)
• Beneficiário: Douglas Borges de Figueiredo
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 11/02805-0 - Determinação dos aminoácidos essenciais e da influência das bases nitrogenadas em cultivo contínuo de Streptococcus pneumoniae sorotipo 14
• Beneficiário: Anne Leticia Silva Ferri
• Modalidade de apoio: Bolsas no Brasil - Mestrado