抗人 DcR3 单克隆抗体的研制及其鉴定【摘要】 目的: 制备抗人 DcR3单克隆抗体, 并鉴定其特异性。方法: 纯化的 His DcR3融合蛋白免疫小鼠, 应用淋巴细胞杂交瘤技术制备抗人 DcR3 mAb 并进行纯化, 纯化后的抗体经 Western blot 和ELISA 方法鉴定其特异性、 Ig 亚型和效价。结果: 获得 5 株稳定分泌抗 DcR3 mAb 的杂交瘤细胞, 均属 IgG1 亚型, 其腹水抗体效价为 1×10-5～1×10-7, Western blot 显示 5 株细胞分泌的 mAb 均可识别 DcR3 蛋白, 其中 1 株可与 SW480 细胞成分反应。结论: 成功建立稳定分泌抗人 DcR3 mAb 的杂交瘤细胞株, 其分泌的抗体特异性强、 效价高, 为讨论 DcR3 在组织中的表达、 分布及研制 ELISA 试剂盒奠定基础。【关键词】 DcR3 单克隆抗体 制备 [Abstract] AIM: To prepare monoclonal antibodies(mAbs) against human DcR3 and identify their characterization. METHODS: BALB/c mice were immunized with purified His DcR3 protein, and mAbs against DcR3 which prepared by hybridoma technique were purified and identified by their specificity, subtype, titers via ELISA and Western blot. RESULTS: Five hybridoma cell lines secreting mAbs against human DcR3 were obtained, which were determined as IgG1 subtype and ascites titers of five mAbs against DcR3 reached 1×10-5-1×10-7. Five mAbs were proved to recognize His DcR3 protein specifically, one of which (1B1) could recognize SW480 cell. CONCLUSION: mAbs against DcR3 with high titers and specificity have been prepared and purified successfully, which laid a foundation for the study of DcR3 expression, distribution in tissu and development of ELISA kit. [Keywords]DcR3; monoclonal antibody; preparation 诱骗受体 3(decoy receptor 3, DcR3, 又称 TR6)为近年新发现的一种可溶性肿瘤坏死因子受体(TNFR)超家族成员, 可竞争性结合肿瘤坏死因子(TNF)成员 LIGHT、 FasL 及 TL1A[1, 2], 但并不介导细胞凋亡, 在细胞的生长、 分化、 死亡以及免疫调节等方面发挥重要作用, 与肿瘤、 自身免...
