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目的 :制备多克隆抗体并初步用于TRP 1抗原表位区的研究 ,为白癜风、恶性黑素瘤的免疫治疗提供实验依据。方法 :在大肠杆菌中表达PRSETA/TRP 1融合蛋白 ,用所获得的蛋白免疫新西兰白兔得到多克隆抗体 ,并用ELISA、Western blot方法进行此抗体的鉴定。结果 :①表达了PRSETA/TRP 1融合蛋白 ;②制备了抗TRP 1B细胞表位区的多克隆抗体 ;③并用多克隆抗体检测了毕赤酵母表达的 6His TRP1蛋白 ,证实此抗体效价高、特异性强。结论 :此抗体可用于TRP 1B细胞表位肽段的进一步研究
OBJECTIVE: To prepare polyclonal antibody and preliminary study of TRP1 epitope region, providing experimental evidence for immunotherapy of vitiligo and malignant melanoma. Methods: The PRSETA / TRP 1 fusion protein was expressed in E. coli, and polyclonal antibody was obtained by immunizing New Zealand white rabbits with the obtained protein. The antibody was identified by ELISA and Western blot. Results: ① PRSETA / TRP 1 fusion protein was expressed; ② polyclonal antibody against TRP 1B was prepared; ③ polyclonal antibody was used to detect the 6His TRP1 protein expressed in Pichia pastoris, Strong specificity. Conclusion: This antibody can be used for further study of TRP 1B epitopes