HPV58型E1^E4蛋白的表达纯化和多克隆抗体的制备

Expression and Purification of HPV58 E1^E4 Fusion Protein and Preparation of Polyclonal Antibody Against HPV58 E1^E4

目的:构建pRSET-A\E1^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 recombined plasmid pRSET-A／EI^E4, and to obtain polyclonal antibodies to El^E4 protein. Methods： The constructed recombined plasmid pRSET-AkEI^E4 was transfected into BL21 （DE3） strain to express the El^E4 protein. The inclusion body was collected and purified through Ni-NTA purification system. Polyclonal antibodies to the El^E4 protein were prepared by inoculating rabbits with purified recombinant protein and purified by ammol/Lonium sulfate. Results： The results of Western blotting showed that the prepared polyclonal antibodies had well specificity to EIAE4 protein of HPV58. Conclusion： The polyclonal antibod- ies to El^E4 protein of HPV58 were obtained successfully, which is helpful to research the function and B cell epitopes of this protein.