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Colorimetric analysis of extracellular vesicle surface proteins based on controlled growth of Au aptasensors
Analyst ( IF 4.2 ) Pub Date : 2021-1-20 , DOI: 10.1039/d0an02080j
Xiaojie Wang 1, 2, 3, 4 , Xinxin Yuan 1, 2, 3, 4 , Kexin Fu 1, 2, 3, 4 , Chang Liu 1, 2, 3, 4 , Lu Bai 2, 3, 4, 5, 6 , Xinchao Wang 2, 4, 7, 8, 9 , Xiaoyue Tan 1, 2, 3, 4 , Yuying Zhang 1, 2, 3, 4
Affiliation  

Protein profiling of extracellular vesicles (EVs) provides important information in both clinical cancer diagnosis and relevant biological research studies. Although a variety of bioanalytical techniques have been investigated for EV characterization, limitations such as time-consuming operations, the requirement of large sample volume and demand for specialized instruments hinder their practical applications. Here, we report a simple and wash-free homogeneous colorimetric assay for sensitive detection of surface proteins on EVs. Au nanoparticles were modified with thiolated aptamers to fabricate aptasensors and incubated with EVs. Upon addition of a Au growth reagent, the solution color changed from light red to blue in the presence of target proteins and became deep red when the targets were absent. Expression of CD63, epithelial cell adhesion molecules (EpCAM), and mucin1 in EVs derived from two breast cancer cell lines (MCF-7 and MDA-MB-231) were compared, showing results consistent with western blotting results. The colorimetric assay achieves a limit of detection (LOD) down to 0.7 ng μL−1 against MCF-7 EVs based on the assessment of EpCAM expression, suggesting its potential to be applied in clinical breast cancer diagnosis.

中文翻译:

基于金适体传感器的受控生长的细胞外囊泡表面蛋白的比色分析

细胞外囊泡(EVs)的蛋白质谱分析为临床癌症诊断和相关生物学研究提供了重要信息。尽管已针对电动汽车的特性研究了多种生物分析技术,但诸如耗时的操作,大量样品的需求以及对专用仪器的需求等限制因素限制了其实际应用。在这里,我们报告了一种简单且免洗的均相比色测定法,用于对电动汽车上的表面蛋白进行灵敏检测。金纳米颗粒用硫醇适体修饰,以制造适体传感器,并与电动汽车孵育。加入Au生长试剂后,在目标蛋白存在的情况下溶液颜色从浅红色变为蓝色,而在没有目标蛋白时溶液变为深红色。CD63的表达 比较了来自两种乳腺癌细胞系(MCF-7和MDA-MB-231)的EV中的上皮细胞粘附分子(EpCAM)和mucin1,显示的结果与Western blotting结果一致。比色法可实现低至0.7 ngμL的检测限(LOD)基于EpCAM表达的评估,针对MCF-7 EV -1,表明其潜力可用于临床乳腺癌诊断。
更新日期:2021-02-02
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