基于量子点纳米吸头的荧光侧流免疫分析法

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES Jove-Journal of Visualized Experiments Pub Date : 2024-06-28 DOI:10.3791/67000
Lingzhi Fan, Yue Luo, Wannian Yan, Huanxing Han, Pengfei Zhang
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引用次数: 0

摘要

量子点又称半导体纳米晶体,是用于生物成像和传感的新型荧光标签。然而,通过费力的纯化程序制备的小尺寸(约 10 纳米)量子点-抗体共轭物在使用侧流免疫分析条检测某些微量疾病标记物时表现出有限的灵敏度。在此,我们介绍一种采用一步乳化蒸发法制备量子点纳米吸附剂(QDNB)的方法。以 C 反应蛋白(CRP)为例,利用制备的 QDNB 制作了一种荧光侧流免疫分析仪,用于检测疾病生物标记物。与单个量子点纳米颗粒不同,量子点纳米吸附剂-抗体共轭物作为免疫测定标签的灵敏度更高,这是因为在一个聚合物复合纳米吸附剂中封装了数百个量子点,从而放大了信号。此外,当 QDNB 与抗体共轭时,QDNB 更大的尺寸有利于更容易地离心分离。基于 QDNBs 的荧光横向流动免疫分析仪已制作完成,并在 15 分钟内测定了样品中的 CRP 浓度。测试结果可在紫外光照射下进行定性评估,并在 15 分钟内使用荧光阅读器进行定量测量。
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Fluorescent Lateral Flow Immunoassay Based on Quantum Dots Nanobeads.

Quantum dots, also known as semiconductor nanocrystals, are novel fluorescent labels for biological imaging and sensing. However, quantum dot-antibody conjugates with small dimensions (~10 nm), prepared through laborious purification procedures, exhibit limited sensitivity in detecting certain trace disease markers using lateral flow immunoassay strips. Herein, we present a method for the preparation of quantum dot nanobeads (QDNB) using a one-step emulsion evaporation method. Using the as-prepared QDNB, a fluorescent lateral flow immunoassay was fabricated to detect disease biomarkers using C-reactive protein (CRP) as an example. Unlike single quantum dot nanoparticles, quantum dot nanobead-antibody conjugates are more sensitive as immunoassay labels due to signal amplification by encapsulating hundreds of quantum dots in one polymer composite nanobead. Moreover, the larger size of QDNBs facilitates easier centrifugation separation when conjugating QDNBs with antibodies. The fluorescent lateral flow immunoassay based on QDNBs was fabricated, and the CRP concentration in the sample was measured in 15 min. The test results can be qualitatively assessed under UV light illumination and quantitatively measured using a fluorescent reader within 15 min.

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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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