On-line version ISSN 1025-0298
PEREZ, José Luis et al. Cuantificación de lipopolisacárido en un proteoliposoma obtenido a partir de la superficie externa de Vibrio cholerae O1. Vaccimonitor [online]. 2007, vol.16, n.1, pp. 15-20. ISSN 1025-0298.
Unlike most of the Gram-negative bacteria, Vibrio cholerae O1 lipopolysacharide (LPS), does not react in KDO assay, a widely used method for the quantification of these molecules. Vibrio cholerae O1 LPS can be quantified by dry weight when it is in pure state, but when it is part of a more complex structure as proteoliposomes, other methods such as strong hydrolysis are required for exposition of KDO molecule. In this paper, a rapid, simple and reproducible method for the quantification of LPS in a proteoliposome obtained from the external surface of Vibrio cholerae O1 El Tor Ogawa is described. The method consists in a Western blot assay using an anti- Ogawa LPS monoclonal antibody followed by the evaluation of the reactive profile by densitometry using a dry weight purified and quantified Ogawa LPS as reference pattern. This method can be applied to any other preparation containing LPS in which KDO method is not applicable
Keywords : Vibrio cholerae 01; LPS; Immunoblot; densitometry.