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Enzyme-Linked Immunosorbent Assay with Monoclonal and Single-Chain Variable Fragment Antibodies Selective to Coplanar Polychlorinated Biphenyls

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journal contribution
posted on 22.02.2012, 00:00 authored by Hideyuki Inui, Tetsuya Takeuchi, Akari Uesugi, Fumito Doi, Mikio Takai, Kosuke Nishi, Shiro Miyake, Hideo Ohkawa
Coplanar polychlorinated biphenyls (Co-PCBs) consisting of non-ortho and mono-ortho-chlorinated PCBs are dioxin-like compounds and cause wide contamination in the environment. To monitor Co-PCB residues, it was attempted to establish an enzyme-linked immunosorbent assay (ELISA) with monoclonal and recombinant antibodies selective to Co-PCBs. When 3,3′,5,5′-tetrachlorobiphenoxybutyric acid (PCBH)–keyhole limpet hemocyanin conjugate was immunized into mice, two monoclonal antibodies, Mab-0217 and Mab-4444, were obtained. 3,3′,5,5′-Tetrachlorobiphenyl (PCB80) was determined with an IC50 value of 2.6 and 0.46 ng mL–1 in ELISA based on Mab-0217 and Mab-4444, respectively. Mab-4444 cross-reacted with Co-PCB congeners, except for PCB77 and PCB81. Mab-0217 reacted with PCB80 and cross-reacted with PCB111. A single-chain variable fragment (scFv) antibody derived from Mab-4444 was produced in recombinant Escherichia coli cells. The scFv antibody showed nearly the same sensitivity toward PCBH as the parent monoclonal antibody in ELISA. These results clearly suggested that Mab-4444 and its scFv antibodies were suitable for monitoring the representative congeners of Co-PCBs.