适用免疫组化的突触素与嗜铬素A单克隆抗体的制备与临床初步评价  

Preparation and clinical evaluation of synaptophysin and chromogranin A monoclonal antibodies against immunohistochemistry

作　　者：闫敏 宋军营 曾华辉 袁永 张钟允 翟晋豫 张振强 刘文弟 YAN Min;SONG Jun- Ying;ZENG Hua-Hui;YUAN Yong;ZHANG Zhong- Yun;ZHAI Jin- Yu;ZHANG Zhen-Qiang;LIU Wen-Di(Chinese Medicine Immunology Laboratory, Henan University of Chinese Medicine, Zhengzhou 450046, Chin)

机构地区：[1]河南中医药大学中医药免疫学实验室,郑州450046 [2]河南省肿瘤病理诊断试剂工程技术研究中心,郑州450001

出　　处：《中国免疫学杂志》2017年第12期1828-1831,共4页Chinese Journal of Immunology

基　　金：2015年度河南省科技创新人才计划(154100510019);河南省基本科研业务费(2014KYYWF-ZZCX3-06;2014KYYWF-ZZCX3-04);2014年度河南中医学院创新团队(2014XCXTD01);河南省高校科技创新人才支持计划(13HASTIT027)

摘　　要：目的:制备突触素与嗜铬素A免疫组化单克隆抗体,期望可初步应用于临床。方法:设计Syn与Cg A融合基因,并构建至表达载体上表达获得融合蛋白。经过抗原免疫、细胞融合后筛选获得目标抗体。通过临床样本对比验证,研究Syn和Cg A两者的特异性,评价相关系数。结果:通过筛选,共获得2株分别针对Syn与Cg A的抗体3D9和4A12。通过对比抗体试剂共同验证19种蜡块组织,统计结果分析,结果符合性较好,相关系数分别达到r=0.989 2与r=0.993 9。结论:该方法成功制备获得了可初步适用于免疫组化的突触素与嗜铬素A抗体,该方法可为免疫学抗体研究提供一定的借鉴意义。Objective： To prepare the synaptophysin and chromogranin A monoclonal antibodies with clinical evaluation. Methods： The fuse gene（ Syn and CgA） was designed and it was constructed on the expression vector p ET-28 a. Then,the fusion protein was purified. After protein immunization,cell fusion and screening,the target antibodies were selected. Specificity study and correlation coefficient of Syn and CgA was evaluated by clinical sample comparison validation. Results： By screening,two antibodies 3 D9 and4 A12,respectively,for Syn and CgA,were obtained. 19 kinds of wax block organization were detected by 3 D9,4 A12 and control antibody（ Leica）. The statistical results were analyzed,the results were in good agreement,and the correlation coefficients were r=0. 9892 and r=0. 993 9,respectively. Conclusion： This method is prepared to obtain the synaptophysin and chromogranin A antibodies successfully and both can be used for immunohistochemistry. This method can also provide some reference for the study of antibody.

关 键 词：突触素 嗜铬素A 单克隆抗体 免疫组化 

分 类 号：R392.3[医药卫生—免疫学]