兔抗2,4-D多克隆抗体的制备

采用水溶性碳化二亚胺法(EDC),将2,4-D与牛血清蛋白(BSA)和卵清蛋白(OVA)共价偶联,分别合成免疫原2,4-D-BSA和包被原2,4-D-OVA,经紫外分光光度计扫描鉴定。用合成的免疫抗原免疫新西兰大白兔,并用合成的包被原进行ELISA试验对抗血清进行定性定量测定,结果表明,获得的抗血清效价达1.6×105。用所制备的抗血清建立间接竞争ELISA方法。优化了ELISA的工作条件,方阵测定确定了包被抗原最佳浓度(5μg/mL),抗血清最佳稀释度(1∶50 000),并建立了ELISA标准工作曲线。工作曲线表明在50~2 000ng/mL浓度范围内呈良好的线性关系,该法检测底限为50 ng/mL。

The preparation of polyclone antibody for 2,4-D residue

2,4-D was chemically linked to BSA or OVA by 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide hydrochloride(EDC) to form immunizeantigen and coating antigen.The complete antigen was determined by UV spectra metric scanning method.Polyclonal antibodies against 2,4-D were produced from rabbits,and the antiserum was determined with ELISA test,which showed that the title of the antibodies was 1∶160 000.Indirect competitive ELISA(icELISA) was established with this antiserum.The most appropriate of coating antigen was 5 μg/mL,and optimal dilution 1∶500 00.The standard curve of icELISA was also established.The curve had a favorable linear relation within the concentration range of 50-2 000 ng/mL and the curve indicated that the lowest detection limit was 50 ng/mL.