抗人细胞角蛋白7(CK7)单克隆抗体的制备及鉴定  

Preparation and Identification of Anti-human Cytokeratin 7(CK7) Monoclonal Antibody

作　　者：曾璇 吴意[2,3] 刘如石[1,2] 曾冰洁 扶春艳 杨玉皎 郑姣[1,2] 钟志宏[1,2] ZENG Xuan;WU Yi;LIU Rushi;ZENG Bingjie;FU Chunyan;YANG Yujiao;ZHENG Jiao;ZHONG Zhihong(School of Medicine,Hunan Normal University,Changsha 410013,China;Immunodiagnostic Reagents Engineering Research Center of Hunan Province,Hunan Normal University,Changsha 410013,China;Department of Clinical Laboratory,the First Affiliated Hospital of Hunan Normal University(Hunan Provincial People’s Hospital),Changsha 410005,China;Maternity Department,Women and Children Healthcare Hospital of Zhuzhou,Zhuzhou 412008,China)

机构地区：[1]湖南师范大学医学院,长沙410013 [2]湖南师范大学免疫诊断试剂湖南省工程研究中心,长沙410013 [3]湖南师范大学附属第一医院(湖南省人民医院)临床检验科,长沙410005 [4]湖南省株洲市妇幼保健院产科,株洲412008

出　　处：《激光生物学报》2022年第3期234-242,共9页Acta Laser Biology Sinica

基　　金：国家自然科学基金青年项目(81702005);湖南省自然科学基金项目(2019JJ50370)。

摘　　要：用KLH偶联合成细胞角蛋白7(CK7)优势表位多肽片段(C14)免疫小鼠,经细胞融合技术制备CK7单克隆抗体(mAb),并收集临床组织进行免疫组化鉴定。C14多肽作为抗原具有良好的免疫反应性。本研究共获得了3株能稳定分泌抗CK7抗体的单克隆细胞株,选择亲和力较好的7A8单克隆抗体进行免疫组化鉴定,结果表明,本研究筛选的7A8单克隆抗体的质量浓度为0.49μg/mL时,与收集的临床标本均有良好的特异性反应,低于医院常用CK7检测试剂盒中抗体质量浓度2~5μg/mL,表现出较高的灵敏度。本研究筛选的7A8单克隆抗体为研发CK7快速诊断试剂提供了关键的原材料。To immunize mouse with KLH-conjugated synthetic polypeptide fragment(C14) of cytokeratin 7(CK7) against the dominant epitope, after cell fusion and being screened, we prepared monoclonal antibodies(mAbs) against CK7, collected clinical tissues for immunohistochemical identification. The results showed that C14 polypeptide has significant immunoreactivity and immunogenicity as an antigen,and three diagnostic monoclonal antibody cell lines that stably secretes antibodies have been obtained. The 7A8 monoclonal antibody with better affinity was selected for immunohistochemical identification, and the antibody concentration was 0.49 μg/mL which is lower than 2~5 μg/mL in the commonly used CK7 detection kits in hospitals,showing high sensitivity. It showed a good specific reaction with the collected clinical specimens, and the key raw material were provided by the screened 7A8 monoclonal antibody for the development of CK7 rapid diagnostic reagents.

关 键 词：细胞角蛋白7(CK7) 免疫组化 单克隆抗体(mAb) 免疫反应性 检测试剂盒 

分 类 号：R392[医药卫生—免疫学]