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Purification of capsular polysaccharide from Streptococcus pneumoniae serotype 23F by a procedure suitable for scale‐up
Author(s) -
Gonçalves Viviane Maimoni M.,
Takagi Mickie,
Lima Rodrigo B.,
Massaldi Hugo,
Giordano Roberto C.,
Tanizaki Martha M.
Publication year - 2003
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20020075
Subject(s) - microbiology and biotechnology , streptococcus pneumoniae , polysaccharide , biology , serotype , ultrafiltration (renal) , streptococcaceae , ethanol precipitation , chromatography , chemistry , biochemistry , antibiotics
Streptococcus pneumoniae is a pathogenic encapsulated bacterium, which causes pneumonia, bacteraemia and meningitis. Capsular polysaccharide conjugated to a carrier protein has been widely used as a vaccine antigen. Serotype 23F is one of the prevalent worldwide pneumococci. A simple and efficient method for capsular polysaccharide serotype 23F purification that can easily be scaled‐up was developed. This method consisted of using culture broth obtained by tangential microfiltration through a 0.22 μ m membrane, broth microfiltrate concentration by tangential ultrafiltration in a 30 kDa spiral membrane, fractional ethanol precipitation (28–60%), nuclease and proteinase treatment, and concentration/diafiltration in a 30 kDa cassette membrane. The final polysaccharide recovery was 89%. The final protein and nucleotide contamination was 1.5% (w/w) and 0.3% (w/w) respectively. The final pure polysaccharide meets the requirements of the World Health Organization and residual proteinase was not found in the final product.