文献类型：期刊文献

英文题名：Quantification of Exosome Based on a Copper-Mediated Signal Amplification Strategy

作者：He, Fang[1];Wang, Jing[1];Yin, Bin-Cheng[1];Ye, Bang-Ce[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biosyst & Microanal, Shanghai 200237, Peoples R China;[2]Zhejiang Univ Technol, Coll Pharmaceut Sci, Collaborat Innovat Ctr Yangtze River Delta Reg Gr, Hangzhou 310014, Zhejiang, Peoples R China;[3]Shihezi Univ, Sch Chem & Chem Engn, Xinjiang 832000, Peoples R China

年份：2018

卷号：90

期号：13

起止页码：8072

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20182505352754);WOS:【SCI-EXPANDED(收录号:WOS:000438008600040)】；

基金：This work was jointly supported by the National Natural Science Foundation of China (Grants 21335003, 21675052, and 21575089), the Fundamental Research Funds for the Central Universities, the Science Fund for Creative Research Groups (Grant 21421004), and the Programme of Introducing Talents of Discipline to Universities (Grant B16017).

语种：英文

外文关键词：Hydrophobicity - Nanoparticles - Cholesterol - Signal processing - Body fluids - Cost effectiveness - Fluorescence - Cost benefit analysis

摘要：Exosomes, a class of small extracellular vesicles, play important roles in various physiological and pathological processes by serving as vehicles for transferring and delivering membrane and cytosolic molecules between cells. Since exosomes widely exist in various body fluids and carry molecular information on their originating cells, they are being regarded as potential noninvasive biomarkers. Nevertheless, the development of convenient and quantitative exosome analysis methods is still technically challenging. Here, we present a low-cost assay for direct capture and rapid detection of exosomes based on a copper-mediated signal amplification strategy. The assay involves three steps. First, bulk nanovesicles are magnetically captured by cholesterol-modified magnetic beads (MB) via hydrophobic interaction between cholesterol moieties and lipid membranes. Second, bead-binding nanovesicles of exosomes with a specific membrane protein are anchored with aptamer-modified copper oxide nanoparticles (CuO NPs) to form sandwich complexes (MB-exosome CuO-NP). Third, the resultant sandwich complexes are dissolved by acidolysis to turn CuO NP into copper(II) ions (Cu2+), which can be reduced to fluorescent copper nanoparticles (CuNPs) by sodium ascorbate in the presence of poly(thymine). The fluorescence emission of CuNPs increases with the increase of Cu2+ concentration, which is directly proportional to the concentration of exosomes. Our method allows quantitative analysis of exosomes in the range of 7.5 x 10(4) to 1.5 x 10(7) particles/yL with a detection of limit of 4.8 X 10(4) particles/mu L in biological sample. The total working time is about 2 h. The assay has the potential to be a simple and cost-effective method for routine exosome analysis in biological samples.