2,4-二氯苯氧乙酸人工抗原合成及多克隆抗体的制备

Synthesis of 2,4-D Artificial Antigen and Preparation of its Polyclonal Antibody

采用碳化二亚胺法,将2,4-二氯苯氧乙酸(2,4-D)分别与牛血清白蛋白(BSA)及卵清蛋白(OVA)偶联,得到完全免疫原2,4-D-BSA和包被原2,4-D-OVA,采用紫外扫描法判定偶联结果 ;用2,4-D-BSA免疫新西兰大耳白兔,采用间接酶联免疫吸附测定法测定多抗血清效价、阻断酶联免疫吸附测定法测定其抑制曲线。结果表明:BSA与2,4-D偶联后,波峰出现右移,且具有2,4-D特征峰,表明偶联成功;2只新西兰大耳白兔血清效价均达到了1∶5.12×105,且B号兔子多抗血清敏感性最好,半数抑制浓度IC50为47.64 ng/m L,表明成功获得了高效价、特异性好、亲和力较高的兔源抗2,4-D多克隆抗体血清,为2,4-D残留免疫学检测方法的建立奠定了良好基础。

2,4-D separately coupling in carrier protein BSA and OVA by using EDC method. The conjugates of 2,4-D-BSA and 2,4-D-OVA were used as a immunogen and coating antigen respectively. UV was used to identify artificial antigen. New Zealand white rabbits were immunized with 2,4-D-BSA. The titers of polyclonal antiserum was detected by indirect ELISA,the sensitivity of polyclonal antiserum was identified by blocking ELISA. The animal immunization results showed that it was successfully coupled when the wave moved to the right after BSA and 2,4-D was coupling. Two rabbits indirect ELISA titers against 2,4-D were above 1 ： 5.12x105 and the IC50 of B rabbit was 47.64 ng/mL,This test was synthesized successfully 2,4-D artificial antigens,2,4-D polyclonal antiserum with high sensibility and specificity was gotten. This polyclonal antibody may lay a foundation for the further 2,4-D monoclonal antibody preparation and the immunology fast detection methods.