Purification of capsular polysaccharide produced by <Emphasis Type="Italic">Haemophilus influenzae</Emphasis> type b through a simple,efficient and suitable method for scale-up |
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Authors: | Mickie Takagi Rodrigo Barbosa Lima Silvia Maria Ferreira Albani Teresa Cristina Zangirolami Martha Massako Tanizaki Joaquin Cabrera-Crespo |
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Affiliation: | 1.Centro de Biotecnologia,Instituto Butantan,S?o Paulo,Brazil;2.Departamento de Engenharia Química,Universidade Federal de S?o Carlos-UFSCar,S?o Carlos,Brazil |
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Abstract: | Haemophilus influenzae type b, an encapsulated bacterium, causes meningitis in infants worldwide. The capsular polysaccharide conjugated to a carrier
protein is effective in the prevention of such infections. The traditional purification process of polysaccharide from bacterial
cultures for vaccine production is based on several selective precipitations with solvents such as: ethanol, phenol, and cationic
detergents. The separations of solid and liquid phases are based on continuous centrifugation in explosion proof installations.
The lipopolysaccharides are separated by ultracentrifugation. A simple and efficient method that can easily be scaled-up was
developed for purification of polysaccharides. The ethanol precipitation was reduced to only two steps. The phenol treatment
was substituted by ultrafiltration and enzymatic digestion. Lipopolysaccharide was removed by ultrafiltration together with
addition of detergent and chelating agent. |
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Keywords: | Polysaccharide purification Tangential ultrafiltration Enzymatic digestion |
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