量子点偶联抗体型夹心免疫传感法检测心肌肌钙蛋白I

高等学校化学学报 ›› 2009, Vol. 30 ›› Issue (10): 1940.

量子点偶联抗体型夹心免疫传感法检测心肌肌钙蛋白I

宋健^1,2 , 范佳², 宋大千³, 毕丽荣⁴, 周广宇⁴, 张皓³, 魏景艳^2,3, 杨柏³

1. 吉林大学电子科学与工程学院, 长春 130012;
2. 吉林大学药学院, 长春 130021;
3. 吉林大学化学学院, 长春 130012;
4. 吉林大学中日联谊医院, 长春 130033

收稿日期:2009-06-24 出版日期:2009-10-10 发布日期:2009-10-10
通讯作者: 魏景艳, 女, 博士, 教授, 博士生导师, 主要从事生物化学和免疫学研究. E-mail: weijy@jlu.edu.cn
基金资助:
国家自然科学基金(批准号: 29875010)和吉林省科技厅科研基金(批准号: 200705380)资助.

Sandwich Immunosensing Method for Measuring Cardiac Troponin I with Quantum Dots-coupled Antibody Against Troponin I

SONG Jian^1,2, FAN Jia², SONG Da-Qian³, BI Li-Rong⁴, ZHOU Gang-Yu⁴, ZHANG Hao³, WEI Jing-Yan^2,3*, YANG Bai³

1. College of Electronic Science and Engineering, Jilin University, Changchun 130012, China;
2. College of Pharmacy, Jilin University, Changchun 130021, China;
3. College of Chemistry, Jilin University, Changchun 130012, China;
4. China-Japan Union Hospital of Jilin University, Changchun 130033, China

Received:2009-06-24 Online:2009-10-10 Published:2009-10-10
Contact: WEI Jing-Yan. E-mail: weijy@jlu.edu.cn
Supported by:
国家自然科学基金(批准号: 29875010)和吉林省科技厅科研基金(批准号: 200705380)资助.

摘要/Abstract

摘要：

将纳米量子点(QD)的放大作用与夹心免疫传感技术相结合, 首次应用量子点标记抗体和表面等离子体共振生物传感器(SPR)对心肌肌钙蛋白I(cTn I)进行特异性定量检测. 利用N-羟基琥珀酰(NHS)和1-乙基-3-(3-二甲氨基丙基)碳二亚胺盐酸盐(EDC)将量子点偶联到cTn I的单克隆抗体2F11上, 再利用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)验证偶联是否成功, 膜印迹法证明标记后的2F11具有良好的生物学和免疫学活性, 最后以蛋白A为基底膜、特异性抗心肌肌钙蛋白I多克隆抗体为第一抗体(捕捉抗体)、QD标记的抗心肌肌钙蛋白I单克隆抗体2F11为第二抗体(检测抗体), 用表面等离子体共振生物传感器构建了对心肌肌钙蛋白I具有特异性的夹心免疫传感法, 并成功用于检测心肌肌钙蛋白I. 本法的检测范围为0.4~15 μg/L, 检出限为0.4 μg/L, 较未标记夹心法和直接法分别提高了约2倍和10倍.

关键词: 心肌肌钙蛋白I; 特异性抗心肌肌钙蛋白I抗体; 夹心免疫传感法; 量子点; 生物传感器

Abstract:

In this study, a novel quantitative sandwich immunoassay for detecting cTn I was developed by using surface plasmon resonance biosensor and quantum dots(QD)-coupled antibody for the first time, which amplified the response of biosensor and so had the sensitivity of this method obviously improved. First quantum dots were coupled to the monoclonal antibody(2F11) against cTn I with N-hydroxysuccinimide(NHS) and 1-ethyl-3-(3-dimethyl-aminopropyl)-carbodimidehydrochloride(EDC), and then the coupled product(QD-2F11) was confirmed by SDS-PAGE and its antibody activity was tested by protein blot. Finally, a sensitive sandwich immunosensing method for measuring cardiac troponin I was developed by surface plasmon resonance biosensor with protein A as an intermediate layer, the specific polyclonal antibody as the first antibody[capture antibody, and QD-labeled-monoclonal antibody as the second antibody(detecting antibody)]. The novel sandwich immunoassay showed the detection range of 0.4 to 15 μg/L and a detection limit of0.4 μg/L, which is higher 2 times than the sandwich immunoassay without using QD-labeled antibody and 10 times than the direct immunoassay for cTn I previously developed, respectively.

Key words: Cardiac troponin I; Specific antibody against cardiac troponin I; Sandwich immunosensing me-thod; Quantum dot; Biosensor

TrendMD:

宋健 , 范佳, 宋大千, 毕丽荣, 周广宇, 张皓, 魏景艳, 杨柏. 量子点偶联抗体型夹心免疫传感法检测心肌肌钙蛋白I. 高等学校化学学报, 2009, 30(10): 1940.

SONG Jian, FAN Jia, SONG Da-Qian, BI Li-Rong, ZHOU Gang-Yu, ZHANG Hao, .... Sandwich Immunosensing Method for Measuring Cardiac Troponin I with Quantum Dots-coupled Antibody Against Troponin I. Chem. J. Chinese Universities, 2009, 30(10): 1940.

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