The haptens 4-{4-[2,2,2-trichloro-1-(4-chloro-phenyl)-ethyl]-phenyl}-butyric acid(DDT-H1),4-[4-(2,2,2-trichloro-1-p-tolyl-ethyl)-phenyl]-butyric acid(DDT-H2) were designed and synthesized on the basis of characteristics part of DDT,and artificial antigens DDT-H1-BSA(immmue antigen),and DDT-H1-OVA.DDT-H2-OVA(coating antigens) were prepared by coupling with the carrier proteins using active ester method and mixture anhydrides method,respectively.The monoclonal antibody(MAb) against DDT was produced by immunity DDT-H1-BSA,cell fusion, screening,cloning. The MAb was prepared from ascetic fluids of Balb/c,which was purified with saturated ammonium sulfate followed by affinity chromatography on protein A.The monoclonal antibody characterized by IgG1 isotype showed a high cross-reactivity to some metabolites.The enzyme linked immunosorbent assay(ELISA) titer of ascites was 1.68×105,and the affinity of antibody(Ka) was 5.238×1011 L?mol-1.An indirect competitive ELISA was founded base on the monoclonal antibody.The results of indirect competitive ELISA showed that the linear range of the calibration curves ranged from 6.6ng/mL to 521.8ng/mL.The method could be applied in the detection of DDT and its metabolites in agriculture products and environment.
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Guangdong Provincial Key Laboratory of Food Quality and Safety,College of Food Science,South China Agricultural University