基于量子点荧光探针的高灵敏蛋白质检测方法

doi:10.3969/j.issn.1000-5641.2016.02.016

华东师范大学学报(自然科学版) ›› 2016, Vol. 2016 ›› Issue (2): 144-152.doi: 10.3969/j.issn.1000-5641.2016.02.016

基于量子点荧光探针的高灵敏蛋白质检测方法

吴思敏^1,2，贾春平²，刘莉芬²，郜晚蕾²，景奉香²，,金庆辉²，张红锋¹，赵建龙^2

1. 华东师范大学生命科学学院, 上海200241；
2. 中国科学院上海微系统与信息技术研究所传感技术联合国家重点实验室, 上海 200050

收稿日期:2015-03-23 出版日期:2016-03-25 发布日期:2016-07-25
通讯作者: 张红锋，女，副教授，硕士研究生，研究方向为肿瘤生物学.  E-mail:hfzhang@bio.ecnu.edu.cn.
作者简介:吴思敏，女，硕士研究生，研究方向为肿瘤生物学. E-mail: wusimin_mail@163.com.
基金资助:
国家重点基础发展计划（2012CB933303）;国家自然科学基金（81101645，61271162，81472751，21405167）；上海市科委项目（12nm0503702，11JC1414400，BRC2012002）; 东南大学生物电子学国家重点实验室开放研究项目

Ultrasensitive detection of protein biomarker using fluorescent quantum dot probes

WU Si-Min^1,2, JIA Chun-Ping², LIU Li-Fen², GAO Wan-Lei², JING Feng-Xiang², Jin-Qing-Hui²， , ZHANG Hong-Feng¹, ZHAO Jian-Long^2

Received:2015-03-23 Online:2016-03-25 Published:2016-07-25

摘要/Abstract

摘要： 通过制备量子点荧光检测探针，构建了一种基于量子点探针和免疫磁珠的蛋白质检测方法，实现了对蛋白肿瘤标志物癌胚抗原（carcinoembryonic antigen, CEA）的高灵敏定量检测. 在该检测体系中，若存在有靶标蛋白，其与量子点检测探针以及捕获探针之间会发生免疫反应形成三明治结构，利用磁力分离器对免疫复合物进行富集后，通过检测富集在磁珠表面的量子点荧光信号，可实现对靶标蛋白的定量. 该方法的检测灵敏度为38 pg/mL,线性范围为0.39～50 ng/mL，临床质控样本检验结果表明，该方法准确度高，可重复性好，可应用于临床样本检测. 该量子点探针检测体系具有灵敏度高、特异性好、样品消耗量低等优点，在疾病早期诊断方面具有广阔的应用前景.

关键词: 量子点, 免疫荧光分析, 免疫磁珠, 肿瘤标志物

Abstract: In this study, based on the unique properties of quantum dots (QDs) and immune magnetic beads, an fluorescent assay was built to detect low concentration carcinoembryonic antigen (CEA). The assay was carried out in a sandwich formation in which two antibodies simultaneously recognized two separate binding sites on the same target CEA. After enrichment with magnetic separator, the fluorescent intensities produced by the QDs, which were confined to the surface of the beads, could be read efficiently to quantify the concentration of the CEA. For this assay, the limit of detection was 38 pg/mL with the linear range was from 0.39 to 50 ng/mL, and it has been proved with high accuracy and repeatability in tumor marker control. With the high sensitivity, specificity and low sample consumption, this method takes a big step to the early diagnosis of diseases.

Key words: quantum dots, immunofluorescence assay, magnetic beads, tumor marker

中图分类号:

吴思敏, 贾春平, 刘莉芬, 郜晚蕾, 景奉香, 金庆辉, 张红锋, 赵建龙. 基于量子点荧光探针的高灵敏蛋白质检测方法[J]. 华东师范大学学报(自然科学版), 2016, 2016(2): 144-152.

WU Si-Min, JIA Chun-Ping, LIU Li-Fen, GAO Wan-Lei, JING Feng-Xiang, Jin-Qing-Hui , ZHANG Hong-Feng, ZHAO Jian-Long-. Ultrasensitive detection of protein biomarker using fluorescent quantum dot probes[J]. Journal of East China Normal University(Natural Sc, 2016, 2016(2): 144-152.

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基于量子点荧光探针的高灵敏蛋白质检测方法

Ultrasensitive detection of protein biomarker using fluorescent quantum dot probes

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