Purification of capsular polysaccharide produced by Haemophilus influenzae type b through a simple, efficient and suitable method for scale-up.
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.
Takagi
Mickie
M
Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil.
Lima
Rodrigo Barbosa
RB
Albani
Silvia Maria Ferreira
SM
Zangirolami
Teresa Cristina
TC
Tanizaki
Martha Massako
MM
Cabrera-Crespo
Joaquin
J
D019149:Bioreactors / Q000382:microbiology; D002498:Centrifugation / Q000379:methods*; D011232:Chemical Precipitation; D000431:Ethanol / Q000737:chemistry; D019741:Haemophilus influenzae type b / Q000737:chemistry* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000737:chemistry / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D014462:Ultrafiltration / Q000379:methods*
D008070:Lipopolysaccharides; D011135:Polysaccharides, Bacterial; D000431:Ethanol