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目的:构建pRSET-AE1^E4原核表达载体,获得E1^E4蛋白的多克隆抗体。方法:在BL21(DE3)细胞中诱导E1^E4融合蛋白表达,收集包涵体经过镍柱纯化后免疫新西兰白兔,获得的抗血清经硫酸铵沉淀法纯化得到抗体。结果:经Western blotting检测表明制备的E1^E4蛋白多克隆抗体特异性较好。结论:成功获得了纯化的E1^E4蛋白抗体,有助于后续HPV58型E1^E4蛋白功能和B细胞抗原表位的研究。
OBJECTIVE: To construct a prokaryotic expression vector pRSET-A E1E4 and to obtain polyclonal antibody against E1E4 protein. Methods: The expression of E1 (superscript E4) fusion protein was induced in BL21 (DE3) cells. The inclusion bodies were purified after purification by nickel column and immunized New Zealand white rabbits. The obtained antiserum was purified by ammonium sulfate precipitation. Results: The results of Western blotting showed that the specificity of the polyclonal antibody of E1 ^ E4 was better. Conclusion: The purified E1 ^ E4 antibody was successfully obtained, which is helpful to study the function of E1 (superscript E) E4 protein and B cell epitopes.