基于多肽抗原的抗人TTC36的单克隆抗体制备及其在免疫组化中的应用  

Preparation of Anti-Human TTC36 Monoclonal Antibody and Its Application in Immunohistochemistry Based on Polypeptides Antigen

作　　者：刘运洪 夏勇武 孙炎[2] 张伟 郭慧娟 姜孝新 LIU Yunhong;XIA Yongwu;SUN Yan;ZHANG Wei;GUO Huijuan;JIANG Xiaoxin(Clinical Laboratory,People’s Hospital of Longhua Shenzhen,Shenzhen 518109,China;Department of Pathology,Third People’s Hospital of Shenzhen,Shenzhen 518109,China)

机构地区：[1]深圳市龙华区人民医院检验科,深圳518109 [2]深圳市第三人民医院病理科,深圳518109

出　　处：《中国细胞生物学学报》2020年第10期1758-1765,共8页Chinese Journal of Cell Biology

基　　金：深圳市龙华区科技创新资金(批准号:2017115);院内博士启动科研项目(批准号:20181125)资助的课题。

摘　　要：为了研究人TTC36的功能,筛选并制备抗人TTC36的单克隆抗体,探讨其在免疫组化检测中的应用。使用化学法合成4种TTC36多肽,将偶联KLH后的多肽免疫Balb/c小鼠,采用杂交瘤方法筛选和制备相应单克隆抗体,使用ELISA、Western blot对单克隆抗体的亚型、效价、特异性进行鉴定和分析,使用免疫组化技术鉴定其性能。结果获得了5株单克隆抗体细胞株,其分泌的抗体能结合原核细胞和真核细胞表达的TTC36蛋白,经免疫组化技术能检测到人肝脏和肾脏中TTC36蛋白表达,并发现肝癌和肾癌组织中的TTC36蛋白表达下降。该研究成功制备了人TTC36的单克隆抗体,通过免疫组化检测了人肝脏和肾脏中TTC36蛋白的表达情况,为TTC36相关疾病的研究、临床诊断、单克隆抗体药物的进一步研究提供基础。This study aimed to investigate the function of anti-TTC36 monoclonal antibodies,especially its application in immunohistochemistry.Four TTC36 polypeptides coupled with KLH were used as antigens to immunize Balb/c mice.The corresponding monoclonal antibodies were screened and prepared by hybridoma method.ELISA and Western blot were used to identify and analyze the subtype,titer and specificity of monoclonal antibodies.The application was investigated with immunohistochemical techniques.Results showed that five monoclonal antibodies recognizing human TTC36 protein were obtained,which could specifically bind to TTC36 protein expressed by prokaryotic cells and eukaryotic cells.These monoclonal antibodies could recognize TTC36 protein in human liver and kidney tissue,which also indicated that TTC36 expression decreased in liver and kidney cancer compared with normal tissue.In conclusion,monoclonal antibodies that bind to human TTC36 have been successfully prepared and can apply in immunohistochemistry,which provides the basis for the clinical diagnosis and drug research of related diseases.

关 键 词：TTC36 多肽 单克隆抗体 免疫组化 

分 类 号：R392[医药卫生—免疫学]