Purification of capsular polysaccharide from Neisseria meningitidis serogroup C by liquid chromatography.
Neisseria meningitidis serogroup C capsular polysaccharide (MenCPS) is an important antigen against meningococcal infection. This paper describes a new purification methodology employing liquid chromatography that resulted in a polysaccharide showing the characteristics recommended by the World Health Organization for vaccine purposes. In this method, steps of the traditional procedure that yield low recovery and use toxic materials were modified. The present process consists in the following steps: (1) continuous flow centrifugation of the culture for removal of the cells; (2) supernatant concentration by tangential filtration (100 kDa cutoff); (3) addition of 0.5% DOC, heating to 55 degrees C during 30 min and tangential filtration (100 kDa cutoff); (4) anion exchange chromatography (Source 15Q) and (5) size exclusion chromatography (Sepharose CL-4B). The polysaccharide C fraction obtained in that way was dialyzed and freeze-dried. The structural identity of the polysaccharide was demonstrated by (1)H-NMR spectrometry.
Departamento de Vacinas Bacterianas, Bio-Manguinhos, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil. Tania@bio.fiocruz.br
Tânia Pinheiro
TP
Pato
Antonio de Pádua R
Ade P
Barbosa
José Godinho
JG
da Silva Junior
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D009345:Neisseria meningitidis / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011135:Polysaccharides, Bacterial / Q000302:isolation & purification*
D011135:Polysaccharides, Bacterial