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

Author(s)

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

• Document type: Journal article
• Source: Brazilian Journal of Chemical Engineering; v. 33, n. 3, p. 435-443, 2016-09-00.

Abstract

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)

• FAPESP's process: 08/05207-4 - Pneumococcal conjugate vaccine: capsular polysaccharide - pneumococcal surface protein A
• Grantee: Martha Massako Tanizaki
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 11/02805-0 - Determination of essential amino acids and influence of nitrogenous bases on the continuous cultivation of Streptococcus pneumoniae serotype 14
• Grantee: Anne Leticia Silva Ferri
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 12/04858-7 - Development of the purification process of the pneumococcal surface protein a from Clade4 (PspA4pro)
• Grantee: Douglas Borges de Figueiredo
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 08/10364-1 - Impact of Streptococcus pneumoniae cultivation strategies on capsular polysaccharide production
• Grantee: Viviane Maimoni Gonçalves
• Support Opportunities: Regular Research Grants