摘要
目的 制备抗人钠碘转运体 (NIS)单克隆抗体。方法 以人NIS蛋白质的二个片段(第 2膜外段和第 14f段 )为抗原免疫小鼠 ,运用杂交瘤技术 ,制备鼠抗人NIS单克隆抗体 (rhNISMAb) ,采用小鼠Ig亚类测定试剂条测定单抗的亚类 ,并通过ELISA、Western blot、免疫组化技术等鉴定其特异性。结果 运用杂交瘤技术成功地制备出能稳定分泌抗人NIS单克隆抗体的细胞系。Western blot分析表明 :此单抗所识别的靶分子的相对分子量主要在 97KD ,免疫组化染色显示 :甲状腺滤泡细胞基底膜上有棕黄色着色。结论 成功地制备出抗人NIS单克隆抗体 ,为NIS抗原 抗体的研究提供了新的工具 ,NIS单克隆抗体不仅可以应用于甲状腺、乳腺等领域的基础研究 ,而且可以用于临床 。
Objective To develop monoclonal antibody against human sodium iodide symporter.Methods The hybridoma cell lines were obtained by fusing spleen cells of Balb/c mice that had been immunized with synthesized NIS peptides.The monoclonal antibody was characterized by ELISA,Western-blot and immunohistochemical staining.Results One hybridoma cell line was developed.This cell line could stably secrete monoclonal antibody which could react specifically to synthesized NIS peptides.The result of Western-blot showed that the monoclonal antibody could recognize molecules of relative molecular mass 97KD in the target cells.Immunostaining with this monoclonal antibody was observed in Graves thyroid tissues.Conclusion Monoclonal antibody to human sodium iodide symporter has been developed and characterized successfully.
出处
《江苏医药》
CAS
CSCD
北大核心
2003年第6期419-421,共3页
Jiangsu Medical Journal
基金
国家自然科学基金资助项目 (3 9970 3 5 1)
关键词
人钠碘转运体
单克隆抗体
杂交瘤技术
电泳印渍检测法
Human sodium iodide symporter Monoclonal antibody Hybridoma Western-blot Immunohistochemical staining
