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Biotechnology and Applied Biochemistry (2003) 37, (283–287) (Printed in Great Britain)

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

Viviane Maimoni M. Gonc¸alves*¹, Mickie Takagi*, Rodrigo B. Lima*, Hugo Massaldi†, Roberto C. Giordano‡ and Martha M. Tanizaki*

*Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil, 1500, 05503-900, São Paulo, SP, Brazil , †Departamento de Desarrollo Biotecnológico y Producción, Instituto de Higiene, Montevideo, Uruguay , and ‡Dep. Engenharia Química, Universidade Federal de São Carlos, Via Washington Luiz, km 235, 13565-905, São Carlos, SP, Brazil

Key words: downstream processing, pneumococcal vaccine, tangential filtration.

Abbreviations used: DOC, sodium deoxycholate; K_d, distribution coefficient; PS, polysaccharide; WHO, World Health Organization.

¹To whom correspondence should be addressed (e-mail vivimaig@butantan.gov.br).

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 µm 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.

Received 28 August 2002/28 November 2002; accepted 6 January 2003

Published as Immediate Publication 6 January 2003, DOI 10.1042/BA20020075

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