抗人t-PSA单克隆抗体的制备及化学发光检测方法的建立  

Preparation of monoclonal antibodies against human t-PSA and establishment of a sandwich CLIA system

作　　者：曹领改 张兰兰 魏德强 CAO Ling-Gai;ZHANG Lan-Lan;WEI De-Qiang(College of Life Science,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学生命科学学院

出　　处：《中国免疫学杂志》2019年第13期1599-1603,共5页Chinese Journal of Immunology

基　　金：中央高校基本科研业务费专项资金(2572019BD03)和中央高校基本科研业务费专项资金(2572014AA19)

摘　　要：目的:制备抗人前列腺癌特异性抗原(t-PSA)单克隆抗体,建立t-PSA的化学发光CLIA检测方法。方法: t-PSA抗原免疫小鼠后,通过细胞融合、HTS筛选、扩大培养及亲和纯化后得到t-PSA单克隆抗体(mAb),测定抗体亲和力、特异性及表位,最后选择抗不同表位的单克隆抗体建立双抗体夹心 CLIA检测方法。结果:获得4株阳性信号较强的抗人t-PSA的mAb(3-21-1、3-26-1、3-40-1、3-41-1)。用3-21-1 mAb-HRP和3-26-1 mAb-HRP建立的CLIA体系检测范围为0.1~100 ng/ml,最低检测限为 0.09 ng/ml,相对偏差均在±5%之内,与肿瘤标志物CEA、AFP、Myo无交叉反应。与罗氏试剂检测t-PSA结果对比,一致性检验KAPPA系数为0.933,相关系数为0.936,两组数据具有较好的一致性和相关性。结论:本研究成功制备了抗人t-PSA mAb,建立了定量检测t-PSA的双抗体夹心CLIA 方法。Objective: To prepare total prostate specific antigen(t-PSA)monoclonal antibody,and establish double antibody sandwich Chemiluminescence immunoassay(CLIA)detecting system for human t-PSA. Methods: Mice were immunized with human t-PSA,and then t-PSA mAb was obtained through cell fusion,HTS screening,expansion culture and affinity purification.Finally the obtained monoclonal antibodies were used to establish the double antibody sandwich CLIA system,which were against different epitopes. Results: 4 human anti-t-PSA(3-21-1,3-26-1,3-40-1,3-41-1)monoclonal antibody were obtained.3-21-1 mAb-HRP and 3-26-1 mAb-HRP were used to establish the double antibody sandwich system with detection range of 0.1-100 ng/ml,sensitivity of 0.09 ng/ml,relative detection deviation of ±5% and not cross-react with the tumor marker AFP,CEA and Myo.Compared with the results of t-PSA detected by Roche reagent,the KAPPA coefficient was 0.933 and the correlation coefficient was 0.936.The two groups of data had good consistency and correlation. Conclusion: The mAbs against human t-PSA have been prepared successfully and the double antibody sandwich CLIA system for quantitative detection of human t-PSA has been established.

关 键 词：人前列腺癌特异性抗原 单克隆抗体 肿瘤标志物 化学发光免疫分析 

分 类 号：R392.11[医药卫生—免疫学] R392-33[医药卫生—基础医学]