Abstract

BALBOA, Julio A et al. Purification of Neisseria meningitidis lipopolysaccharide starting from a collateral fraction of the process of the antimeningococcal vaccine, VA-MENGOC-BC® production process. Vaccimonitor [online]. 2008, vol.17, n.1, pp.17-26. ISSN 1025-028X.

The work aimed at purifying lipopolysaccharides (LPS) of Neisseria meningitidis from a collateral fraction of the antimeningococcal BC vaccine, VAMENGOC-BC®. production process, the supernatant obtained from the ultra centrifugation stage during the proteins extraction process of the meningococcus outer membrane. The purification was carried out by precipitation 80% ethanol, protein extraction with 90% phenol from 65-70 ºC and fractional ultra centrifugation at 105 000 g. Three lots of LPS were obtained, in total 1.069 g, with a content of proteins, nucleic acids and sialic acid in respect to the LPS of 0.5%, 0.3% and 2.2% (m/m) respectively. The assessment by chromatography showed a high molecular integrity. with constant valves of reproducible distribution (Kd 0.36 - 0.38) and a possible sialic acid association to the LPS. Homogeneity was observed in the electrophoretic profile of the three lots and a high endotoxic activity. The purified LPS was mainly identified as the inmunotype L3,7,9. The purification procedure used allows making use of a collateral fraction of the vaccine production process, it is scalable, it does not include chromatographic methods and makes easy the obtainment of large quantity of LPS of Neisseria meningitidis, wihich it is non available on the market, with high purity and high endotoxic activity.

Keywords : lipopolysaccharide; lipooligosaccharide; purification; Neisseria meningitidis; meningococcal vaccine.

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