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目的制备5-氟尿嘧啶(5-FU)免疫原并制备其多克隆抗体。方法采用化学合成法对5-FU进行修饰制备5-氟尿嘧啶-1-基乙酸半抗原(5-FUAA),并经碳酰二亚胺盐法将其分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)连接以制备免疫原。采用5-FU与BSA结合的免疫原(5-FUAA-BSA)免疫BALB/c小鼠制备多克隆抗体,5-FU与OVA结合物(5-FUAA-OVA)包被酶标板经间接ELISA检测抗血清效价,并采用ELISA与Western blot法检测抗血清的特异性。结果成功合成半抗原5-FUAA,并分别制备5-FUAA-BSA和5-FUAA-OVA完全抗原。将5-FUAA-BSA免疫BALB/c小鼠制备的免疫血清效价达1∶1 280 000,且该多克隆抗体能特异地结合5-FU半抗原。结论成功制备了5-FU的多克隆抗体。
Objective To prepare 5-fluorouracil (5-FU) immunogen and prepare its polyclonal antibody. Methods The 5-fluorouracil-1 -ylacetate hapten (5-FUAA) was modified by 5-FU using chemical synthesis method. The 5-fluorouracil- Albumin (OVA) to prepare the immunogen. BALB / c mice were immunized with 5-FU and BSA-conjugated immunogen (5-FUAA-BSA) to prepare polyclonal antibody. 5-FU and OVA conjugate (5-FUAA- The antiserum titer was detected and the specificity of antiserum was tested by ELISA and Western blot. Results The hapten 5-FUAA was successfully synthesized and 5-FUAA-BSA and 5-FUAA-OVA complete antigen were prepared respectively. BALB / c mice immunized with 5-FUAA-BSA had an immune serum titer of 1: 1 280 000 and the polyclonal antibody specifically bound to the 5-FU hapten. Conclusion The polyclonal antibody of 5-FU was successfully prepared.