Purification of capsular polysaccharide from Streptococcus pneumoniae serotype 23F by a procedure suitable for scale-up.
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.
Gonçalves
Viviane Maimoni M
VM
Centro de Biotecnologia, Instituto Butantan, Av Vital Brasil, 1500, 05503-900, São Paulo, SP, Brazil. vivimaig@butantan.gov.br
Takagi
Mickie
M
Lima
Rodrigo B
RB
Massaldi
Hugo
H
Giordano
Roberto C
RC
Tanizaki
Martha M
MM
D016667:Bacterial Capsules / Q000737:chemistry* / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000302:isolation & purification; D010450:Endopeptidases / Q000302:isolation & purification; D009711:Nucleotides / Q000302:isolation & purification; D010865:Pilot Projects; D011135:Polysaccharides, Bacterial / Q000302:isolation & purification; D013296:Streptococcus pneumoniae / Q000737:chemistry* / Q000302:isolation & purification*; D014462:Ultrafiltration / Q000379:methods*
D001426:Bacterial Proteins; D009711:Nucleotides; D011135:Polysaccharides, Bacterial; D010450:Endopeptidases