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2,4-D人工抗原合成及2,4-D类农药特异性多克隆抗体的研制

Keywords: 2,4-D,半抗原,免疫抗原,酶联免疫吸附测定(ELISA),抗体

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Abstract:

通过改良EDC法,将2,4-DB、2,4-D分别与载体蛋白(BSA、OVA)偶联合成了2,4-D的免疫原2,4-DB-BSA、2,4-D-BSA和包被原2,4-DB-OVA、2,4-D-OVA。免疫抗原测定结果表明,所得紫外吸收光谱,具有载体蛋白BSA和半抗原2,4-DB或2,4-D的特性,最大吸收峰为282nm。合成2,4-DB-BSA、2,4-D-BSA免疫抗原的结合比分别为1∶24、1∶20,合成的2,4-DB-OVA、2,4-D-OVA包被原的结合比分别为1∶10、1∶8。上述两种免疫原分别免疫新西兰白雄兔,获得2,4-D类农药多克隆抗体。其中免疫2,4-DB-BSA的两只兔PBD1和PBD2的效价较高,分别是1∶102400和1∶51200。选择效价较高的PBD1多克隆抗体进一步优化筛选出抗原抗体的工作浓度,2,4-D-OVA包被原3μg·mL-1,PBD1多克隆抗体1∶12800。经测定,该多克隆抗体对目标检测物2,4-D、2,4-Dbutylester敏感度较高,抑制中浓度(I50)、检测限(I20)分别为,2,4-D:179.8ng·mL-1,1.09ng·mL-1;2,4-Dbutylester:3010.0ng·mL-1,1.53ng·mL-1。对2,4-D识别的特异性也较强。另外测定了制备2,4-D多克隆抗体对几种2,4-D类似结构化合物交叉特异性反应,2,4-DB最强,2,4-D次之,之后依次

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