人白介素24单克隆抗体的制备与鉴定

Preparation and identification of monoclonal antibodies against interleukin 24

目的:制备抗白细胞介素24(Interleukin 24,IL-24)的单克隆抗体并对其生物学特性进行鉴定。方法:应用分子生物学技术,构建含人IL-24编码序列的原核表达载体,表达并纯化了人IL-24融合蛋白。用纯化人IL-24融合蛋白免疫Balb/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0进行融合,用间接ELISA方法筛选杂交瘤细胞,有限稀释法进行亚克隆,采用免疫印迹及免疫组化染色对抗体的特异性进行了鉴定。结果:获得3株能稳定分泌抗IL-24单抗的杂交瘤细胞,分别命名为1G2,3F4,4A6。通过免疫印迹及免疫组化染色检测显示,这3株单抗均能够与IL-24特异性结合。结论:成功制备抗IL-24单克隆抗体,为进一步探索IL-24在抗肿瘤中的作用机制奠定了基础。

Objective： To prepare and identify the monoclonal antibody against interleukin 24（IL-24）.Methods： Prokaryotic expressing vector containing human IL-24 gene was constructed by molecular biology techniques.Then human IL-24 fusion protein was expressed and purified.Splenocytes from the Balb/c mice immunized by human IL-24 fusion protein were fused with myeloma cells SP2/0 for producing hybridoma.Hybridoma cell line secreting antibodies against human IL-24 was determined by indirect ELISA and limiting dilution assay.The specificity of monoclonal antibodies against human IL-24 were evaluated with Western blot and immunohistochemistry staining.Results： Three hybridoma cell lines stably secreting monoclonal antibodies against human IL-24 were developed and named 1G2,3F4 and 4A6 respectively.By Western blot and immunohistochemistry staining,the results indicated that all monoclonal antibodies raised could specifically react with human IL-24.Conclusion： The success in production of IL-24 monoclonal antibody provides a foundation for studying the function of IL-24 in anti-tumor experiments.