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| Preparation and Evaluation of DDT Monoclonal Antibody |
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| DOI： |
| KeyWord:dichlorodiphenyltrichloroethane(DDT) monoclonal antibody ELISA metabolites |
| Author | Institution |
| YANG Xing-xing,BI Si-yuan,ZHU Hai,GU Da-yong* |
1.广东省深圳市易瑞生物技术有限公司;2.深圳市检验检疫科学研究院 |
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| Abstract: |
| 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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