可溶性fms样酪氨酸激酶1时间分辨荧光免疫分析法的建立  

Establishment of time-resolved fluoroimmunoassay for soluble fms-like tyrosine kinase 1

作　　者：吴鸿峰 邓昊 翁孔燕 黄颖而 陈佳灵 李志雄 吴英松 郝文波 WU Hong-feng;DENG Hao;WENG Kong-yan;HUANG Yin-er;CHEN Jia-ling;LI Zhi-xiong;WU Ying-song;HAO Wen-bo(Institute of Antibody Engineering,Southern Medical University School of Laboratory Medicine and Biotechnology,Guangzhou 510515,China;Guangzhou Darui Biotechnology Co.,Ltd.,Guangzhou 510705,China;School of Laboratory Medicine and Biotechnology,The Key Laboratory of Antibody Engineering of Guangdong Province Higher Education Institutes,Guangzhou 510515,China)

机构地区：[1]南方医科大学检验与生物技术学院抗体工程研究所,广州510515 [2]广州市达瑞生物技术股份有限公司,广州510705 [3]广东省高等学校抗体工程重点实验室检验与生物技术学院,广州510515

出　　处：《现代免疫学》2022年第5期375-380,共6页Current Immunology

摘　　要：为建立基于磁性微球可溶性fms样酪氨酸激酶1(soluble fms-like tyrosine kinase 1,sFlt-1)的时间分辨荧光免疫分析法(sFlt-1 time-resolved fluoroimmunoassay, sFlt-1-TRFIA),用磁珠偶联抗sFlt-1单克隆抗体,Eu^(3+)(铕)标记抗sFlt-1单克隆抗体,采用双抗体夹心法建立基于磁珠的sFlt-1-TRFIA。结果显示,自制试剂的检测灵敏度为10.78 pg/mL,线性范围10.78~25 000 pg/mL;样品的回收率为98.23%~107.71%,分析内变异系数(intraassay coefficient of variation, CV)为5.29%~6.50%,分析间变异系数(intreassay coefficient of variation, CV)为6.55%~8.23%;与电化学发光法的检测结果比较,相关系数r^(2)值为0.971 9。由此,基于磁性微球建立的sFlt-1-TRFIA灵敏度、特异度以及准确度等均符合临床应用要求,有望用于临床血清样本中sFlt-1浓度的检测。This study aimed to establish a time-resolved fluoroimmunoassay based on magnetic microspheres soluble fms-like tyrosine kinase 1(sFlt-1),the monoclonal antibody against sFlt-1 was labeled with magnetic beads and the monoclonal antibody against sFlt-1 was labeled with Eu^(3+).sFlt-1 time-resolved fluoroimmunoassay(sFlt-1-TRFIA)based on magnetic beads was established by the double-antibody sandwich method.The results showed that the sensitivity of the method was 10.78 pg/mL,and the linear range was 10.78~25000 pg/mL.The average recovery rate of the sample was 98.23%~107.71%,the intraassay coefficient of variation(CV)was 5.29%~6.50%,and the intreassay CV was 6.55%~8.23%.The expression level of serum sFlt-1 was compared with the results of chemiluminescent immunoassay,and the correlation coefficient(r^(2)-value)was 0.9719.In conclusion,the sensitivity,specificity and accuracy of sFlt-1-TRFIA based on magnetic microspheres all meet the requirements of clinical application,and this method can be used to detect the concentration of sFlt-1 in patient serum sample.

关 键 词：磁性微球 可溶性fms样酪氨酸激酶1 时间分辨荧光免疫分析法 先兆子痫 

分 类 号：R392-33[医药卫生—免疫学]