Zhou Qing, Rahimian Ali, Son Kyungjin, Shin Dong-Sik, Patel Tushar, Revzin Alexander
Department of Biomedical Engineering, University of California, Davis, CA, United States.
Department of Biomedical Engineering, University of California, Davis, CA, United States; Department of Medical and Pharmaceutical Sciences, Sookmyung Women's University, Seoul, South Korea.
Methods. 2016 Mar 15;97:88-93. doi: 10.1016/j.ymeth.2015.10.012. Epub 2015 Oct 21.
Exosomes are small (50-100 nm in diameter) vesicles secreted from various mammalian cells. Exosomes have been correlated with tumor antigens and anti-tumor immune responses and may represent cancer biomarkers. Herein, we report on the development of an aptamer-based electrochemical biosensor for quantitative detection of exosomes. Aptamers specific to exosome transmembrane protein CD63 were immobilized onto gold electrode surfaces and incorporated into a microfluidic system. Probing strands pre-labeled with redox moieties were hybridized onto aptamer molecules anchored on the electrode surface. In the presence of exosomes these beacons released probing strands with redox reporters causing electrochemical signal to decrease. These biosensors could be used to detect as few as 1×10(6) particles/mL of exosomes, which represents 100-fold decrease in the limit of detection compared to commercial immunoassays relying on anti-CD63 antibodies. Given the importance of exosome-mediated signal transmission among cells, our study may represent an important step towards development of a simple biosensor that detects exosomes without washing or labeling steps in complex media.
外泌体是由各种哺乳动物细胞分泌的小囊泡(直径50 - 100纳米)。外泌体已与肿瘤抗原和抗肿瘤免疫反应相关联,可能代表癌症生物标志物。在此,我们报告了一种基于适配体的电化学生物传感器的开发,用于定量检测外泌体。将外泌体跨膜蛋白CD63特异性的适配体固定在金电极表面,并整合到微流控系统中。用氧化还原基团预标记的探针链与锚定在电极表面的适配体分子杂交。在外泌体存在的情况下,这些信标释放出带有氧化还原报告基团的探针链,导致电化学信号降低。这些生物传感器可用于检测低至1×10⁶个颗粒/毫升的外泌体,与依赖抗CD63抗体的商业免疫测定相比,检测限降低了100倍。鉴于外泌体介导的细胞间信号传递的重要性,我们的研究可能代表了朝着开发一种简单生物传感器迈出的重要一步,该传感器能够在复杂介质中无需洗涤或标记步骤即可检测外泌体。