JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:51 min • August 31st, 2026
Take lentiviral vectors containing HIV-1 p24 antigen, a protein marker used for detection.
Add a detergent to lyse and release p24, then serially dilute the sample.
Take a microplate coated with anti-p24 monoclonal antibodies and a blocking agent to prevent non-specific binding.
Add the sample dilutions and p24 standards as references.
Incubate to allow p24 interaction with the antibodies via a specific epitope, then remove unbound components.
Incubate with anti-p24 polyclonal antibodies that bind to different epitopes on p24. Remove any unbound antibodies.
Add peroxidase-linked antibodies that bind to the polyclonal antibodies. Remove unbound antibodies.
Add a peroxidase substrate that converts into a colored product.
Add an acid to inactivate peroxidase and stabilize the product.
Measure the absorbance corresponding to the color intensity, which is proportional to the p24 concentration.
Generate the standard graph and estimate the sample concentrations to determine the viral titer.
Coat a high-binding 96-well plate with 100 microliters of monoclonal anti-p24 antibody at a dilution of one to 1,500 according to NIH AIDS vaccine program for HIV-1 p24 antigen capture assay kit. Incubate the pla
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