Meningococcal Antigen Typing System Development and Application to the Evaluation of Effectiveness of Meningococcal B Vaccine and Possible Use for Other Purposes

Figure 2

Schematic representation of the MATS ELISA method [15]. Bacteria from overnight cultures on agar chocolate plates are suspended in Mueller-Hinton broth and lysed with a detergent (Empigen BB 5%) added to a final volume of 1/11 and inactivated for 1 hour at 45°C in a water bath; bacterial lysates are added to three different ELISA microwell plates coated with polyclonal rabbit antibodies raised against the single vaccine components fHbp, NHBA, and NadA; the antigens are captured from the suspension to the plate. Plates are then incubated for 1 hour at 37°C with biotinylated rabbit polyclonal antibodies against each of the antigens, washed, incubated with streptavidin-HRP, and developed with the OPD substrate. Relative potency is calculated by interpolating the regression curve of the unknown sample versus that of a reference strain added to each plate. Adapted with permission from [15].