t-PAIC单克隆抗体制备及磁微粒化学发光免疫检测  

Preparation of Monoclonal Antibody of t-PAIC and Its Magnetic Particle Chemiluminescence Immunoassay

作　　者：王兢磊 孔飞志 党智笙 钱纯亘 聂立波 WANG Jinglei;KONG Feizhi;DANG Zhisheng;QIAN Chungen;NIE Libo(College of Life Science and Chemistry,Hunan University of Technology,Zhuzhou Hunan 412007,China;Shenzhen Yhlo Biotech Co.,Ltd.,Shenzhen Guangdong 518116,China)

机构地区：[1]湖南工业大学生命科学与化学学院,湖南株洲412007 [2]深圳市亚辉龙生物科技股份有限公司,广东深圳518116

出　　处：《包装学报》2023年第4期59-67,共9页Packaging Journal

基　　金：湖南省自然科学基金资助项目(2020JJ6102);湖南省教育厅科学研究基金资助项目(19A144);湖南省大学生创新创业训练计划资助项目(202211535029)。

摘　　要：以组织型纤溶酶原激活剂-抑制剂复合物(t-PAIC)为免疫原制备和筛选了一对特异性单克隆抗体,并建立了一种检测人体血浆t-PAIC的磁微粒化学发光免疫检测方法。通过与参比方法比对,选定了反应速度更快的一步法作为反应模式,然后对t-PAIC检测方法的检测性能进行了评估。结果显示:该方法的空白限为0.43 ng/mL,检出限为0.91 ng/mL;线性范围为0.91~100ng/mL;重复性CV小于4%,精密度CV小于3%;样本回收率在100%~110%之间;常见干扰物对检测结果无明显影响。构建了t-PAIC检测方法的参考区间,健康成年男性参考范围为1.85~15.91 ng/mL,健康成年女性参考范围为1.90~9.43 ng/mL。t-PAIC检测方法反应快速,在血栓疾病诊断中有重要的应用价值。A pair of specific monoclonal antibodies were prepared and screened by using tissue type plasminogen activator inhibitor complex(t-PAIC)as immunogen,and then a magnetic particle chemiluminescence immunoassay method was established for the detection of human plasma t-PAIC.In comparison with the reference method,the one-step method with a faster reaction speed was finally selected as the reaction mode.Then the performance of the method was estimated,the results showed that the limit of blank(LOB)was 0.43 ng/mL,the limit of detection(LOD)was 0.91 ng/mL,and the linear range was 0.91~100 ng/mL.The coefficient of variance(CV)of repeatability was less than 4%,and the CV of precision was less than 3%.The sample recovery rate was between 100%~110%,and the common interfering substances had no obvious influence on the detection results.Finally,the reference interval of the method was constructed.The reference range for healthy adult males was 1.85~15.91 ng/mL,and that of 1.90~9.43 ng/mL for healthy adult women.The method has the advantage of rapid reaction,which is of important application value in the diagnosis of thrombotic diseases.

关 键 词：单克隆抗体 化学发光 人组织型纤溶酶原激活剂-抑制剂复合物 吖啶酯 

分 类 号：R392.11[医药卫生—免疫学] R446.62[医药卫生—基础医学]