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以BaP为原料,经硝化和还原引入氨基合成半抗原(oABaP),再采用碳化二亚胺(EDC)法将半抗原分别与牛血清白蛋白(BSA)及卵清蛋白(OVA)偶联,合成人工抗原(BSA-oABaP)和包被抗原(OVA-oABaP),通过紫外扫描法和聚丙酰胺凝胶电泳(SDS-PAGE)法进行鉴定;免疫BALB/c纯系小鼠,采用间接ELISA测定小鼠多抗血清效价、阻断ELISA测定其抑制价。结果表明:紫外扫描结果显示BSA-oABaP的波峰发生右移,说明偶联成功;SDS-PAGE法显示BSA的泳动速度大于BSA-oABaP,证明偶联成功;免疫的6只小鼠血清抗体的效价均达到1:103以上,其中6号小鼠的血清效价最高,半数抑制浓度IC50为22.14μg/mL,表明获得了具有良好特异性的多克隆抗体血清,这为苯并芘ELISA快速检测试剂盒的研制奠定了基础。
BaP was used as raw material to synthesize hapten (oABaP) by nitration and reduction. Coupled with bovine serum albumin (BSA) and ovalbumin (OVA) by carbodiimide (EDC) The synthetic antigen (BSA-oABaP) and coating antigen (OVA-oABaP) were identified by ultraviolet scanning and polyacrylamide gel electrophoresis (SDS-PAGE). BALB / c mice were immunized with indirect ELISA Mouse multi-antiserum titer, blocking ELISA determination of its inhibition. The results showed that the peak of BSA-oABaP was shifted to the right by UV scanning, indicating that the coupling was successful. The SDS-PAGE showed that the mobility of BSA was higher than that of BSA-oABaP and the coupling was successful. The titer reached above 1: 103. Among them, the serum titer of No. 6 mice was the highest, and the IC50 of half inhibitory concentration was 22.14 μg / mL, which indicated that polyclonal antibody sera with good specificity was obtained. The development of test kit has laid a foundation.