单 位：
大连水产学院水产养殖系;国家海洋环境监测中心;大连海事大学环境科学与工程学院
关键词：
DDA;DDT;抗DDA抗体;酶联免疫吸附实验
摘 要：
制备抗DDA抗体,建立间接竞争酶联免疫吸附实验(Indrect Competitive Enzyme-linked Immunosorbent Assay,idc-ELISA)方法,以测定海水样品和海洋贝类中的有机氯农药滴滴涕(DDT)代谢产物。采用碳二亚胺法,将半抗原DDA分别与载体蛋白牛血清白蛋白(Bovine serumalbumin,BSA)和卵清蛋白(Ovalbumin,OVA)偶联,得到完全抗原。以DDA-BSA为抗原,免疫BALB/c小鼠,制备抗DDA抗体。研究表明,5只鼠产生的抗体对DDA都能产生灵敏的特异性免疫反应,以DDA-OVA为包被抗原,用5#抗体建立间接竞争ELISA方法检测海水样品和海洋贝类中的有机氯农药DDT代谢产物DDA,最低检出限为:10.0ng·mL-1,海水加标样平均回收率122%,批间变异系数30%,贝类加标样平均回收率为131%,批间变异系数22%。
译 名：
Preparation and Caracterization of Polyclonal Antibody Against DDA
作 者：
LIU Ren-yan1,2,3, DONG Yu-hua4, XU Dao-yan2,3, LIANG Yu-bo2,3 (1.Environmental Science & Engineering College, Dalian Maritime University, Dalian 116026,China; 2.National Marine Environmental Monitoring Center, Dalian 116023, China; 3.SOA Key Lab of Coastal Environment and Ecosystem Research, Dalian 116023, China; 4. Department of Aquaculture, Dalian Fisheries University, Dalian 116023, China)
关键词：
DDA; DDT; polyclonal antibody; enzyme-linked immunosorbent assay (ELISA)
摘 要：
Organochlorine pesticide 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane(DDT) is a persistent and broad toxic organochlorine insecticide. DDT is slowly degraded in tissues mainly to 2,2-bis(4-chlorophenyl)acetic acid(DDA). DDT and its decomposition products are ubiquitous micropollutants in seawater and shellfish. This study presents a novel method of preparing the polyclonal antibody against DDA to investigate the levels of DDT and DDA in seawater and shellfish using indirect competitive enzyme-linked immunosorbent assay (idc-ELISA). Under the presence of N,N-dicyclohexylcarbodiimide, the complete antigen can be obtained by coupling the hapten (DDA) with bovine serum albumin (BSA) and ovalbumin (OVA), respectively. The female BALB/c mice were immunized using DDA-BSA as immunogen to prepare the polyclonal antibody against DDA and its characterization was studied. The results indicated that the antibodies produced from five mice immunized possessed highly sensitive and specific immune reaction for DDA. Using DDA-OVA as coating antigen, the 5# antibody was employed to develop the idc-ELISA method to investigate the concentrations of DDA in seawater and shellfish. The limit of detection (LOD) of the method was 10.0 ng·mL-1. The average recovery and coefficient of variation (CV) were 122% and 30% for seawater, and 131% and 22% for shellfish, respectively.
