滴滴涕代谢物DDA单克隆抗体的制备及其酶联免疫检测方法的建立

Preparation of monoclonal antibody to DDT metabolite DDA and its application in analysis of DDT residue in shellfish

为建立一种快速检测生物体中有机氯农药滴滴涕(DDT)代谢物DDA的酶联免疫检测(Enzyme-linked immunosorbant assay,ELISA)方法,利用碳二亚胺(EDC)法将DDT代谢产物半抗原DDA(2,2-双(4-氯苯基)乙酸)与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,分别作为免疫抗原和包被抗原.采用杂交瘤技术制备特异性抗DDA单克隆抗体,同时建立DDA间接竞争ELISA检测方法,并应用到海产贝类DDA的检测分析中.结果显示,本研究制备抗体所建立的ELISA方法在优化条件下对DDA的半抑制浓度IC50和检测限(Limit of Detection,LOD)分别为110 ng/mL和7.5ng/g,具有高灵敏度.该抗体同DDT其他几种同系物的交叉反应率都较低,只对DDA具有较强的特异性.本研究所制备抗体能够满足海产贝类DDT代谢物DDA的检测.

This study aimed to establish a rapid enzyme-linked immunosorbent assay（ELISA） method to detect DDA, a metabolite of organochlorine pesticide DDT. Hapten of 2, 2-bis-（4-chlorophenyl） acetic acid（DDA） was coupled to bovine serum albumin（BSA） and ovalbumin（OVA） respectively to be used as immunogen and coating antigen. Hybridoma technology was used to prepare specific monoclonal antibody to DDA. An indirect competitive ELISA（idc-ELISA） for determination of DDA was also established and performed on detection of DDA in marine shellfish. The results showed the IC50 and limit of detection（LOD） values under the optimized ELISA conditions were 110 ng/mL and 7.5 ng/g respectively, with high sensitivity. The antibody exhibited low cross reactivity with some related DDT isomersand high specificity to DDA only. The antibody prepared in this study can be applied into the detection of DDA, a metabolilite of DDT in shellfish.