睫状神经营养因子单克隆抗体制备  

Establishment of the monoclonal antibodies against ciliary neurotrophic factor

作　　者：邓小华[1] 周芸[2] 曾瑞云[2] 蒋春雷[1] 路长林[1] 

机构地区：[1]第二军医大学基础医学部神经生物学教研室,上海200433 [2]上海第二医科大学医学检验重点实验室,上海200025

出　　处：《第二军医大学学报》1999年第8期560-562,共3页Academic Journal of Second Military Medical University

摘　　要：目的：制备睫状神经营养因子（ｃｉｌｉａｒｙ ｎｅｕｒｏｔｒｏｐｈｉｃ ｆａｃｔｏｒ， Ｃ Ｎ Ｔ Ｆ）单克隆抗体。方法：将 Ｃ Ｎ Ｔ Ｆ免疫的 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠脾细胞和 Ｓ Ｐ２／０ 骨髓瘤细胞融合，经 ３ 次克隆化培养，获得分泌 Ｃ Ｎ Ｔ Ｆ单克隆抗体的杂交瘤细胞株；然后进行体内诱生获取含大量 Ｃ Ｎ Ｔ Ｆ单克隆抗体的腹水和血清；再进行 Ｃ Ｎ Ｔ Ｆ单克隆抗体鉴定和效价测定。结果：获得５ 株分泌 Ｃ Ｎ Ｔ Ｆ单克隆抗体的杂交瘤细胞株，抗体亚型为 Ｉｇ Ｇ２ａ；腹水和血清中该抗体效价分别为 １∶１０７ ～１∶１０８ 和 １∶１０６；该抗体能特异性地显示 Ｓ Ｄ大鼠脑内表达 Ｃ Ｎ Ｔ Ｆ的神经元，其中１ 株单克隆抗体能抑制 Ｃ Ｎ Ｔ Ｆ促进体外培养新生 Ｓ Ｄ 大鼠骨骼肌细胞增殖的功能。结论：制备成功高效且特异的 Ｃ Ｎ Ｔ Ｆ单克隆抗体，为 Ｃ Ｎ Ｔ Ｆ结构、功能和机制的研究提供了有效工具。Objective: To prepare the monoclonal antibodies against ciliary neurotrophic factor(CNTF). Methods: Cell lines of hybrids secreting monoclonal antibodies against CNTF were established by fusing SP2/0 myeloma cells with spleen cells of BALB/c mice immunized with CNTF and cloning 3 times; asitic fluid and serum containing abundant monoclonal antibodies against CNTF were obtained by in vivo induction; then identification of the monoclonal antibodies against CNTF and detection of antibody titer were undergone. Results: Five cell lines of hybrids secreting monoclonal antibodies against CNTF were established. Subtypes of these antibodies were all IgG 2a . Antibody titers in asitic fluid and serum were 1∶10 7~1∶10 8 and 1∶10 6, respectively. These monoclonal antibodies showed specificity in demonstrating neurons secreting CNTF in CNS, and one kind of them inhibited the enhancing effect of CNTF on the proliferation of neonal SD rats skeletal muscle cells in culture. Conclusion: The monoclonal antibodies against CNTF with high activity and specificity have been established successfully, and will be a useful tool in the studies of CNTF's structure,function and mechanism

关 键 词：神经营养因子 单克隆抗体 CNTF 

分 类 号：R392－33[医药卫生—免疫学]