JoVE Encyclopedia of Experiments
Biological Techniques
0 views • 2:57 min • July 8th, 2025
To elicit an immune response, antigens bind to the antibodies that are specific for the endocytic receptors present on the surface of dendritic cells, or DCs — the antigen-presenting cells — which help in the uptake of antigens.
The formation of an antibody-antigen complex is a crucial step and can be accomplished in vitro by chemically coupling the antigen to a DC-targeting monoclonal antibody.
To verify the efficiency of the conjugation process, begin with an ELISA plate precoated with the captured antibodies. Treat the plate with a blocking solution to prevent the non-specific binding of the antigens to the coated surface.
Add chemically crosslinked, antibody-antigen conjugates to the plate, and incubate. The specific epitope of the antigen interacts with the captured antibody's paratope forming an antibody-antigen-antibody complex.
Next, overlay the plate with a third set of antibodies linked to an enzyme. These antibodies bind specifically to the primary antibodies present in the conjugate, enabling its detection.
Add a buffer containing the chromogenic substrate to visualize the conjugates. The enzyme attached to the antibody reacts with the substrate, generating the colored
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