【摘要】 目的:纯化原核表达的JEV E蛋白并制备其多克隆抗体。方法:IPTG诱导大肠杆菌表达JEV E蛋白,Ni-NTA亲和层析纯化。将纯化后的JEV E蛋白免疫新西兰兔,制备多克隆抗血清。琼脂糖双向扩散实验及间接ELISA检测抗体效价。结果:获得浓度1.85mg/ml、纯度92%的JEV E蛋白,并制备了JEV E蛋白兔多克隆抗体血清。琼脂糖双向扩散实验显示多克隆抗体血清1∶16稀释时仍能观察到沉淀线,间接ELISA法显示1∶200稀释时具有较高效价。结论:纯化了JEV E蛋白,并制备出了多克隆抗体。更多还原

【Abstract】 Objective: To purify JEV E protein expressed in E-coli and prepare its polyclonal antibody.Methods: JEV E protein expressed in E-coli induced by IPTG was purified by Ni-NTA chromatography and was utilized to immune New-Zealand rabbits to achieve blood serum of polyclonal antibody.Doubel immunodiffusion test and indirect ELISA experiment were used to detect antibody titer.Results: The concentration of JEV E protein was 1.85mg/ml and the purity was 92%.When the achieved polyclonal antibody was attenuated according to 1∶16,precipitation line was still found in doubel immunodiffusion test.The polyclonal antibody had high titer when it was attenuated according to 1∶200.Conclusion:JEV E protein and its polyclonal antibody were prepared.更多还原