Objective To generate a specific antibody for perfluorooctanoic acid (PFOA) based on antigen-antibody specific binding, which can be used to develop an immunoassay for on-site fast detection of PFOA.
Methods At first, a PFOAbovine serum albumin (BSA) complete antigen was synthesized by carbodiimide method. Then an varible domain of heavy china of heavy-chain antibody (VHH) phage display library was used for isolating positive clones. The DNA region of the specific antibodies by polymerase chain reaction amplification was ligated to pET22b-Fc vector, and the antibody was expressed using a prokaryotic expression system. The specificity of the antibody was confirmed by indirect competitive enzyme-linked immunosorbent assay (ELISA) assay.
Results The complete antigen PFOA-BSA was synthesized in a coupling ratio of 8. Using the VHH antibody library, 7 positive clones were isolated after four rounds of screening. Five antibodies containing human Fc fragment were obtained with the molecular weights in line with our expectations. Finally, one VHH antibody was verified by indirect competitive ELISA assay showing specific binding to PFOA.
Conclusion A specific VHH antibody for PFOA is obtained, which might be used to develop immunoassays for fast detecting PFOA in various media.
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