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To develop a PFOA antibody that can be integrated into a magnetic particle based immunoassay or other suitable format of rapid screening assay that is cost effective, high throughput, and user friendly.
Approach: Perfluorooctanoic acid is a small molecule that, by itself, cannot elicit an immuno-response to produce antibodies. We will generate PFOA hapten that can attach to large molecules to produce an immuno-response. Rabbits will be immunized with the PFOA immunogen and rabbit serum will be collected when good titer and specificity is shown toward PFOA. The screening assay will be developed using either magnetic particle (tube) or plate format enzyme linked immunoassay (ELISA). Antibodies will be purified from serum and covalently bound to magnetic particles. Perfluorooctanoic acid hapten will be conjugated to horse radish peroxidase (HRP) as a signal generator. Perfluorooctanoic acid present in the samples will compete with the PFOA-HRP for the binding site of the antibody. The more PFOA in the sample the less signal will be generated. When signal strength is compared with signal strengths generated by a known set of PFOA standards, the amount of PFOA in the samples can be computed. An instrumental method such as q-TOF-LC-MS-MS will be used to correlate the screening assay results for the purpose of validation. Environmental samples, such as water and soil, as well as food samples, such as meat and milk, will be utilized for matrix effect determination.