Abstract:
Exosomes are nanovesicles secreted by cells, which play a crucial role in various pathological processes. Exosomes have shown great promise as tumor biomarkers because of the abundant secretion during tumor formation. The development of a convenient, efficient, and cost-effective method for simultaneously enriching and detecting exosomes is of utmost importance for both basic research and clinical applications. In this study, an aptamer-functionalized magnetic Ti3C2 composite material (Fe3O4@Ti3C2@PEI@DSP@aptamer@FAM-ssDNA) is prepared for the simultaneous enrichment and detection of exosomes. CD63 aptamers are utilized to recognize and capture the exosomes, followed by magnetic separation. The exosomes are then released by cleaving the disulfide bonds of DSP. Compared to traditional methods, Fe3O4@Ti3C2@PEI@DSP@aptamer@FAM-ssDNA exhibited superior efficiency in enriching exosomes while preserving their structural and functional integrity. Detection of exosome concentration is achieved through the fluorescence quenching of Ti3C2 and the competitive binding between the exosomes and a fluorescently labeled probe. This method exhibited a low detection limit of 4.21 × 104 particles mL-1, a number that is comparable to the state-of-the-art method in the detection of exosomes. The present study demonstrates a method of simultaneous enrichment and detection of exosomes with a high sensitivity, accuracy, specificity, and cost-effectiveness providing significant potential for clinical research and diagnosis.
摘要:
外泌体是细胞分泌的纳米囊泡,在各种病理过程中起着至关重要的作用。由于在肿瘤形成过程中分泌丰富,外泌体已显示出作为肿瘤生物标志物的巨大前景。一个方便的发展,高效,同时富集和检测外泌体的具有成本效益的方法对于基础研究和临床应用都至关重要。在这项研究中,制备了一种适体功能化的磁性Ti3C2复合材料(Fe3O4@Ti3C2@PEI@DSP@aptamer@FAM-ssDNA),用于同时富集和检测外泌体。CD63适体用于识别和捕获外泌体,其次是磁力分离。然后通过切割DSP的二硫键释放外泌体。与传统方法相比,Fe3O4@Ti3C2@PEI@DSP@aptamer@FAM-ssDNA在富集外泌体方面表现出优异的效率,同时保持其结构和功能完整性。通过Ti3C2的荧光猝灭和外来体与荧光标记的探针之间的竞争性结合来实现外来体浓度的检测。该方法表现出4.21×104颗粒mL-1的低检测限,该数字与检测外来体的最新方法相当。本研究证明了一种高灵敏度同时富集和检测外来体的方法,准确度,特异性,和成本效益为临床研究和诊断提供了巨大的潜力。