甲状腺转录因子-1单克隆抗体的制备及其应用价值分析  

Preparation of Monoclonal Antibody of Thyroid Transcription Factor-1 and Its Application Value

作　　者：陈旭[1,2] 张路遥 刘玲玲 翟晋豫 张会娟 CHEN Xu;ZHANG Luyao;LIU Lingling;ZHAI Jinyu;ZHANG Huijuan(Department of Pathology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,China;Department of Pathology and Pathophysiology,Basic Medical School,Zhengzhou University,Zhengzhou 450052,China;Department of Endocrinology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,China;Henan Celnovte Biotechnology Company Limited,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院病理科,河南郑州450052 [2]郑州大学基础医学院病理学与病理生理学系,河南郑州450052 [3]郑州大学第一附属医院内分泌科,河南郑州450052 [4]河南赛诺特生物技术有限公司,河南郑州450001

出　　处：《肿瘤基础与临床》2019年第5期369-374,共6页journal of basic and clinical oncology

摘　　要：目的制备甲状腺转录因子-1(TTF-1)单克隆抗体,为临床工作及后续研究奠定基础。方法以聚合酶链反应扩增目的基因,并构建TTF-1/pET32a重组质粒,诱导表达融合蛋白,经镍琼脂糖亲和层析柱洗脱目的蛋白,并将洗脱液经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测蛋白纯度,经过免疫学评价合格后,进行小鼠体内免疫,效价评价后通过细胞融合筛选阳性细胞株,制备分泌TTF-1单克隆抗体,并对得到的抗体进行免疫组化验证。结果获得能稳定分泌TTF-1单克隆抗体的杂交瘤细胞株,且其产生的抗体与TTF-1抗原具有良好的免疫学反应。结论成功制备出TTF-1单克隆抗体,为进一步提高相关肿瘤的诊断率提供了新的试剂。Objective To prepare monoclonal antibody of thyroid transcription factor-1(TTF-1),and to lay the foundation for clinical work and follow-up study.Methods The target gene was amplified by polymerase chain reaction and cloned into pET32a vector.The protein expression was induced,then the fusion protein was purified by Ni-chelating affinity chromatography.After determining the molecular weight of the purified protein by sodium dodecyl sulfate polyacrylamide gel electrophoresis,the band was excised from gel for protein identification by enzyme linked immunosorbent assay method.BALB/c mice were immunized with purified fusion protein,and hybridoma cells were obtained by cell fusion.The stability of the obtained hybridoma cells and the specificity of TTF-1 monoclonal antibody were identified.Results One hybridoma cell line which could secrete TTF-1 monoclonal antibody stably was obtained.This monoclonal antibody had good immunological reaction with TTF-1 antigen.Conclusion The monoclonal antibody of TTF-1 is prepared successfully,which would provide a new material for improvement of the diagnostic rate in related tumors.

关 键 词：甲状腺转录因子-1 单克隆抗体 酶联免疫吸附实验 免疫组化 

分 类 号：R730.23[医药卫生—肿瘤]