高等学校化学学报

高等学校化学学报 ›› 2003, Vol. 24 ›› Issue (12): 2161.

• 论文 • 上一篇    下一篇

用多克隆抗体构建的夹心免疫传感器检测心肌肌钙蛋白I

魏景艳1,2, 牟颖1, 许海波3, 胡鑫1, 宋大千4, 刘霞4, 张寒琦4, 金钦汉4, 罗贵民1   

  1. 1. 吉林大学分子酶学工程教育部重点实验室;
    2. 吉林大学药学院, 长春 130023;
    3. 吉林大学第二医院, 长春 130041;
    4. 吉林大学化学学院, 长春 130023
  • 收稿日期:2002-12-26 出版日期:2003-12-24 发布日期:2003-12-24
  • 通讯作者: 罗贵民(1943年出生),男,教授,博士生导师,主要从事抗体酶研究.E-mail:gmluo@mai.ljlu.edu.cn E-mail:gmluo@mai.ljlu.edu.cn
  • 基金资助:

    国家自然科学基金(批准号:29875010);吉林省卫生厅科研基金(批准号:98047)资助

Measurement of Cardiac Troponin I by Sandwich Immunobiosensor Prepared with the Polyclonal Antibody

WEI Jing-Yan1,2, MU Ying1, XU Hai-Bo3, HU Xin1, SONG Da-Qian4, LIU Xia4, ZHANG Han-Qi4, JIN Qin-Han4, LUO Gui-Min1   

  1. 1. Key Laboratory of Molecular Enzymology and Engineering of Ministry of Education;
    2. College of Pharmacy, Jilin University, Changchun 130023, China;
    3. The Second College of Clinical Medicine Affiliated to Jilin University, Changchun 130041, China;
    4. College of Chemistry, Jilin University, Changchun 130023, China
  • Received:2002-12-26 Online:2003-12-24 Published:2003-12-24

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被引次数

13

摘要： 用表面等离子体子共振生物传感器构建对心肌肌钙蛋白I特异性的免疫传感器检测心肌肌钙蛋白I,并建立两种检测方法:直接法的最低检测限为2.5μg/L,基于传感膜上的夹心免疫法的灵敏度为0.5μg/L,检测范围为0.5～20μg/L,批内及批间精密度分别为3.5%～4.9%,6.1%～7.4%;用夹心法及国外试剂盒对40名健康献血者和20例急性心肌梗死患者血清心肌肌钙蛋白I水平进行检测,两者符合率为95%.

关键词: 心肌肌钙蛋白I, 特异性抗心肌肌钙蛋白I多克隆抗体, 夹心免疫分析, SPR传感器, 心肌梗死

Abstract: Cardiac and skeletal muscle troponin I(cTnI, sTnI) were purified from human left ventricular tissue and skeletal muscle tissue by affinity chromatographic method, respectively. The specific polyclonal antibody to cTnIwas prepared by immuned pure New Zeal and rabbit with the purified cTnIand purified by nitrocellulose membrance filtration-affinity chromatography with sTnI. Aspecific immunobiosensor for cTnIwas developed by using surface plasmon resonance(SPR) biosensor with biotin as an intermediate layer and with the biotinylated-specific polyclonal antibody as capture antibody and used in a direct detection of cTnIin sera with the lowest detection limit of 2.5 μg/L. Amore sensitive sandwich immunobiosensor for cTnIwas developed by using biotinylated-specific polyclonal antibody as capture antibody(the first antibody) and polyclonal antibody as the second antibody. The sandwich immunoassay showed an excellent sensitivity of 0.5μg/Land the detection range of 0.5-20 μg/Land within-run variation of 3.5%-4.9% and between-run variation of 6.1%-7.4%. The cTnI′s in the sera of 40 healthy donors and 20 patients with acute myocardiaLInfarction(AMI) were monitored by using the sandwich immunoassay and the Lifesign MI Troponin Itest device, respectively and it was found that the coincidence of the two assays was 95%, indicating that the developed sandwich immunoassay with SPRbiosensor is suitable in clinical determination.

Key words: Cardiac troponin I, Polyclonal antibody to cardiac troponin I, Sandwich immunoassay, Surface plasmon resonance biosensor, Acute myocardiaLInfarction

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