Resumo

CUELLO, Maribel et al. SDS-PAGE and densitometric analysis to determine concentration of lipopolysaccharide from Neisseria meningitidis serogroups A, W₁₃₅ and X . Vaccimonitor [online]. 2013, vol.22, n.3, pp. 36-42. ISSN 1025-028X.

For several years, Finlay Institute, in collaboration with the "Norwegian Institute of Public Health", is working in a project to obtain a vaccinal candidate from outer membrane vesicles (OMV) of N. meningitis serogroups A, W₁₃₅ and X. To establish the quality specifications dealing with lipopolysaccharides (LPS), main cause of the pirogenicity of vaccines, is a necessity in this project. This work aims at establishing the conditions of SDS-PAGE to be used as a method for the quality control of the OMV as active pharmaceutical ingredient obtained from N. meningitidis serogroups A, W₁₃₅ and X. The immunotypes of LPS were determined in studies as well as their concentrations as contaminants of OMV. SDS-PAGE gels at 15% and LPS samples from different serogroups were used; these gels were tinted with silver, specific for LPS, and the results were analyzed in a densitometer GS-800 (Bio-Rad) controlled by the program "Quantity One". As a result, LPS profile curves for each serogroup were obtained and the concentration of residual LPS in the OMV of different serogroups of N. meningitidis was quantified. As conclusion, the methodology to determine the concentration of LPS from N. meningitidis serogroups A, X and W₁₃₅ in samples of OMV with SDS-PAGE was established, in addition the established quality specifications for this parameter were confirmed.

Palavras-chave : LPS; Neisseria meningitidis; SDS-PAGE; OMV; vaccines.

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