Detecting HIV early is essential in providing in optimal patient care. p24 is a component of the HIV capsule which is secreted in high levels in the blood during early stages of HIV-1 infection. As such, it can be used for early detection of the disease.
In order to develop an immunoassay to detect p24 in serum, a pair of antibodies would be needed for a sandwich ELISA; one for capture of the desired protein in a mix of multiple other proteins and another antibody to detect the captured protein. Mice were immunized with a recombinant p24 protein and antibody clones generated using hybridoma development. However, in order for antibodies to work together well in a sandwich ELISA or similar capture-detection system, they need to bind to different regions of the protein, far enough away from each other so they will not interfere. To select for such antibodies among the multitude of hybridomas after fusion, we screened using a test matrix to optimize capture of the protein as well as detection of the protein when captured. When using paired antibodies from the same species, it is important to directly label the detection antibody to avoid complications with using a secondary antibody that could detect the capture antibody as well.
Below is a sample HIV-1 p24 ELISA standard curve from our catalog number PM-6585 for capture and PM-6335-biotin for detection. For more information about developing antibody pairs for your assay, contact our Custom Antibody Experts.