| 摘要: |
| 目的：制备睫状神经营养因子(ciliary neurotrophic factor,CNTF)单克隆抗体。方法：将CNTF免疫的BALB/c小鼠脾细胞和SP2/0骨髓瘤细胞融合，经3次克隆化培养，获得分泌CNTF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CNTF单克隆抗体的腹水和血清；再进行CNTF单克隆抗体鉴定和效价测定。结果：获得5株分泌CNTF单克隆抗体的杂交瘤细胞株，抗体亚型为IgG2a；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SD大鼠脑内表达CNTF的神经元，其中1株单克隆抗体能抑制CNTF促进体外培养新生SD大鼠骨骼肌细胞增殖的功能。结论：制备成功高效且特异的CNTF单克隆抗体，为CNTF结构、功能和机制的研究提供了有效工具。 |
| 关键词: 睫状神经营养因子 抗体，单克隆 |
| DOI： |
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| Establishment of the monoclonal antibodies against ciliary neurotrophic factor |
| 路长林,蒋春雷,曾瑞云,周芸,邓小华 |
| () |
| Abstract: |
| 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 IgG2a. Antibody titers in asitic fluid and serum were 1∶107-1∶108 and 1∶106, 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 |
| Key words: ciliary neurotrophic factors antibodies, monoclonal |
