Enzyme-Linked Immunosorbent Assay with Monoclonal and Single-Chain Variable Fragment Antibodies Selective to Coplanar Polychlorinated Biphenyls
Enzyme-Linked Immunosorbent Assay with Monoclonal and Single-Chain Variable Fragment Antibodies Selective to Coplanar Polychlorinated Biphenyls
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DOI：
10.1021/jf203585b
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发表时间：
2012-02-22
影响因子：
6.1
通讯作者：
Ohkawa, Hideo
中科院分区：
文献类型：
--
作者：
Inui, Hideyuki;Takeuchi, Tetsuya;Ohkawa, Hideo
共面多氯联苯（Co-PCBs）是由非邻位和单邻位氯代PCBs组成的一类二恶英类化合物，在环境中造成广泛的污染。为了监测Co-PCB的残留，尝试建立一种酶联免疫吸附试验（ELISA）与单克隆抗体和重组抗体的选择性Co-PCB。将3，3 '，5，5'-四氯联苯氧基丁酸（PCBH）-钥孔血蓝蛋白偶联物免疫小鼠，获得单克隆抗体Mab-0217和Mab-4444。在基于Mab-0217和Mab-4444的ELISA中，3，3 '5，5'-四氯联苯（PCB 80）的IC 50值分别为2.6和0.46 ng mL（-1）。Mab-4444与Co-PCB同系物发生交叉反应，但PCB 77和PCB 81除外。Mab-0217与PCB 80发生反应，与PCB 111发生交叉反应。在重组大肠杆菌细胞中产生来自Mab-4444的单链可变片段（scFv）抗体。在ELISA中，scFv抗体显示出与亲本单克隆抗体几乎相同的对PCBH的敏感性。这些结果清楚地表明，单克隆抗体-4444及其单链抗体适用于监测的代表性同源物的Co-PCBs。
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.