Purification of capsular polysaccharide from Streptococcus pneumoniae serotype 23F by a procedure suitable for scale-up

Biotechnol Appl Biochem. 2003 Jun;37(Pt 3):283-7. doi: 10.1042/BA20020075.

Abstract

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 microm 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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Bacterial Capsules / chemistry*
  • Bacterial Capsules / isolation & purification*
  • Bacterial Proteins / isolation & purification
  • Endopeptidases / isolation & purification
  • Nucleotides / isolation & purification
  • Pilot Projects
  • Polysaccharides, Bacterial / isolation & purification
  • Streptococcus pneumoniae / chemistry*
  • Streptococcus pneumoniae / isolation & purification*
  • Ultrafiltration / methods*

Substances

  • Bacterial Proteins
  • Nucleotides
  • Polysaccharides, Bacterial
  • Endopeptidases