JoVE Encyclopedia of Experiments
Immunology
0 views • 5:42 min • July 8th, 2025
Begin with a passivated assay plate coated with rabies immunogenic glycoprotein-specific monoclonal antibodies and a blocking agent to prevent non-specific interactions.
Introduce serially diluted reference and test rabies vaccines with decreasing glycoprotein content. Seal the plate to prevent contamination.
During incubation, immunogenic glycoproteins interact with coated monoclonal antibodies via a specific antigenic site.
Post-incubation, remove the film, aspirate the supernatant, and wash the plate.
Overlay with enzyme-conjugated glycoprotein-specific monoclonal antibodies that bind to a different antigenic site on the glycoprotein, forming an immunocomplex.
Add a solution containing a substrate and a chromogen. When the substrate interacts with the enzyme, it causes the oxidation of the chromogen, changing the solution color.
Add a stopping solution to inactivate the enzymes.
Measure the absorbance and generate a reference curve using the reference vaccine values.
Identify the region where there is a linear relationship between absorbance and the glycoprotein content of the reference vaccine dilutions. Then, extrapolate the glycoprotein content of the test vaccine dilutions b
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