DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14
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...
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Brazilian Society of Chemical Engineering
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doaj-44ff97f739e44ce1821768f9e07f1bdf2020-11-25T01:36:24ZengBrazilian Society of Chemical EngineeringBrazilian Journal of Chemical Engineering1678-438333343544310.1590/0104-6632.20160333s20150140S0104-66322016000300435DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14R. T. ZanardoA. L. S. FerriD. B. FigueiredoS. KraschowetzJ. Cabrera-CrespoV. M. GonçalvesAbstract 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66322016000300435&lng=en&tlng=enPneumococcal vaccinePurification strategySodium dodecyl sulfateTrichloroacetic acidAnion exchange chromatography |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
R. T. Zanardo A. L. S. Ferri D. B. Figueiredo S. Kraschowetz J. Cabrera-Crespo V. M. Gonçalves |
spellingShingle |
R. T. Zanardo A. L. S. Ferri D. B. Figueiredo S. Kraschowetz J. Cabrera-Crespo V. M. Gonçalves DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14 Brazilian Journal of Chemical Engineering Pneumococcal vaccine Purification strategy Sodium dodecyl sulfate Trichloroacetic acid Anion exchange chromatography |
author_facet |
R. T. Zanardo A. L. S. Ferri D. B. Figueiredo S. Kraschowetz J. Cabrera-Crespo V. M. Gonçalves |
author_sort |
R. T. Zanardo |
title |
DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14 |
title_short |
DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14 |
title_full |
DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14 |
title_fullStr |
DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14 |
title_full_unstemmed |
DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14 |
title_sort |
development of a new process for purification of capsular polysaccharide from streptococcus pneumoniae serotype 14 |
publisher |
Brazilian Society of Chemical Engineering |
series |
Brazilian Journal of Chemical Engineering |
issn |
1678-4383 |
description |
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. |
topic |
Pneumococcal vaccine Purification strategy Sodium dodecyl sulfate Trichloroacetic acid Anion exchange chromatography |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66322016000300435&lng=en&tlng=en |
work_keys_str_mv |
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