人白介素24单克隆抗体的制备与鉴定
刘世海;李荣华;王东阳;王丽娴;夏海滨
【期刊名称】《现代肿瘤医学》 【年(卷),期】2011(019)009
【摘要】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 antihody 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 hyhridoma. 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
人白介素24单克隆抗体的制备与鉴定
