Detecting RSV Protein P with Monoclonal Antibodies

Example 6

The assay was made for determining the maximum dilution of the monoclonal antibodies RSV anti-P from the hybridomas 2E6/D2 and 6H5/H1, which allow the detection of the viral antigen using ELISA. For this, the same indirect ELISA technique was used of the example 6. The well was activated with 50 ng of the purified antigen and the anti-P antibodies 2E6/D2 or 6H5/H1 were used in dilutions 1:2, starting from the concentration of work (3.4 μg/ml) to the dilution 11 in PBS/FBS 10%. In FIG. 3 is observed that to all the dilutions, which were used in the assay, the anti-P 2E6/D2 and 6H5/H1 antibodies are able of detecting the RSV protein P. The anti-P RSVH102 antibody, catalogue number #AB94965, of Abcam, was also able of detecting in all the dilutions the RSV protein P, but it was less efficient than the anti-P 6H5/H1 antibody.

The negative control included on this assay, correspond to a well which does not contain sample (protein P), it was blocked with PBS/FBS 10%, primary antibody was not added (anti-P 2E6/D2 or anti-P 6H5/H1) and it contains only the mouse anti-IgG antibody conjugated with HRP.