Improvement in the purification process of the capsular polysaccharide from Haemophilus influenzae type b by using tangential ultrafiltration and diafiltration.
Capsular polysaccharide produced by Haemophilus influenzae b (Hib) is the main virulent agent and used as the antigen in the vaccine formulation. In this study, an improved process of polysaccharide purification was established based on tangential flow ultrafiltration using detergents (cocamidopropyl betaine and sodium deoxycholate), two selective ethanol precipitations steps, and extensive enzymatic hydrolysis as strategy. The relative purity (RP) related to protein and nucleic acids were 122~263 and 294~480, respectively, and compatible with the specifications established by the World Health Organization for Hib vaccine, RP≥100. These results make this process simple, cheaper, efficient, environmentally friendly, and prone to be scaled up.
Instituto Butantan-Centro de Biotecnologia, Av. Vital Brasil, 1500, 05503-900 São Paulo, Brazil.
Silvia Maria Ferreira
SM
Albani
Mateus Ribeiro
MR
da Silva
Mickie
M
Takagi
Joaquin
J
Cabrera-Crespo
D016667:Bacterial Capsules / Q000096:biosynthesis / Q000302:isolation & purification*; D005947:Glucose / Q000494:pharmacology; D019741:Haemophilus influenzae type b / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D014462:Ultrafiltration / Q000379:methods*
D005947:Glucose