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采用碳化二亚胺法(EDC法)将2,4-D与牛血清白蛋白(BSA)及卵清蛋白(OVA)偶联,合成免疫原2,4-D-BSA和包被原2,4-D-OVA,采用紫外扫描(UV)和聚丙烯酰胺凝胶电泳(SDS-PAGE)法进行鉴定;用2,4-D人工免疫抗原(2,4-D-BSA)免疫BALB/c纯系小鼠,采用间接ELISA测定多抗血清效价、阻断ELISA鉴定其抑制。结果表明:BSA与2,4-D偶联后,波峰出现右移,表明偶联成功;SDS-PAGE电泳中BSA的泳动速度大于2,4-D-BSA,表明2,4-D已与BSA成功偶联;6只小鼠血清效价均达到了1:1.28×104,且1号小鼠多抗血清敏感性最好,半数抑制IC50为72.48ng/mL,表明成功获得了高效价、特异性好、亲和力较高的鼠源抗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的基础。
2,4-D was coupled with bovine serum albumin (BSA) and ovalbumin (OVA) by carbodiimide method (EDC method) to synthesize 2,4-D-BSA and coated original 2, 4-D-OVA and identified by UV-PAGE and SDS-PAGE. Immunized BALB / c mice with 2,4-D artificial immune antigen (2,4-D-BSA) The pure mice were tested for anti-serum titer by indirect ELISA and blocked by ELISA. The results showed that the peak of BSA was shifted to the right after coupling with 2,4-D, indicating that the coupling was successful. The mobility of BSA in SDS-PAGE was higher than that of 2,4-D-BSA, indicating that 2,4-D Was successfully coupled with BSA. The serum titer of all the 6 mice reached 1: 1.28 × 104, and the sensitivity of multi-antiserum in No.1 mice was the best, with the IC50 of 72.48ng / mL, indicating that the titer of high titer , Good specificity and high affinity murine anti-2,4-D polyclonal antibody serum, which laid a good foundation for the establishment of 2,4-D residual immunological detection method.