On-line version ISSN 1025-0298
VALMASEDA, Tania et al. Determination of the optimal assay conditions for an ELISA to detect serum IgG antibodies against Pseudomonas aeruginosa O11 lipopolysaccharide. Vaccimonitor [online]. 2004, vol.13, n.3, pp. 18-26. ISSN 1025-0298.
An indirect quantitative ELISA was developed for the evaluation of the immune response elicited by Pseudomona aeruginosa vaccine candidates, and for obtaining hyperimmune immunoglobulins to be used in the medical treatment. Standard and control sera were made, and the optimal LPS coating dilution of the polystyrene plates, and the sample and conjugate dilutions were established. A 1.5 μg/mL concentration of O11 LPS in PBS, incubated for 16-20 h at 4°C were found to be the best coating conditions. The blocking step was unnecessary. A 1:100 dilution for the serum samples and 1:3000 for the anti-human IgG:HRP conjugate were selected. Accurate quantitative determinations were possible in the lineal range of the standard curve.
Keywords : Pseudomonas aeruginosa; immunoglobulin; lipopolysaccharide; ELISA; standardization.