State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, P. R. China.
School of Life Sciences, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, Shandong 250117, P. R. China.
Anal Chem. 2023 Feb 21;95(7):3606-3612. doi: 10.1021/acs.analchem.2c04280. Epub 2022 Dec 24.
Exosome-based liquid biopsy technologies play an increasingly prominent role in tumor diagnosis. However, the simple and sensitive method for counting exosomes still faces considerable challenges. In this work, the CD63 aptamer-modified DNA tetrahedrons on the gold electrode were used as recognition elements for the specific capture of exosomes. Partially complementary DNA probes act as bridges linking trapped exosomes and three AuNP-DNA signal probes. This clover-like structure can tackle the recognition and sensitivity issues arising from the undesired AuNP aggregation event. When cancerous exosomes are present in the system, the high accumulation of methylene blue molecules from DNA-AuNP nanocomposites on the surface of the electrode leads to an intense current signal. According to the results, the aptasensor responds to MCF-7 cell-derived exosomes in the concentration range from 1.0 × 10 to 1.0 × 10 particles·μL, with the detection limit of 158 particles·μL. Furthermore, the aptasensor has been extended to serum samples from breast cancer patients and exhibited excellent specificity. To sum it up, the aptasensor is sensitive, straightforward, less expensive, and fully capable of receiving widespread application in clinics for tumor monitoring.
基于外泌体的液体活检技术在肿瘤诊断中发挥着越来越重要的作用。然而,用于计数外泌体的简单而灵敏的方法仍然面临着相当大的挑战。在这项工作中,金电极上修饰有 CD63 适体的 DNA 四面体被用作识别元件,用于外泌体的特异性捕获。部分互补的 DNA 探针作为桥接物,将捕获的外泌体与三个 AuNP-DNA 信号探针连接起来。这种三叶草状结构可以解决由于 AuNP 聚集导致的识别和灵敏度问题。当系统中存在癌源性外泌体时,电极表面上的 DNA-AuNP 纳米复合材料中的亚甲基蓝分子大量积累,导致电流信号增强。根据结果,该适体传感器在 1.0×10 至 1.0×10 个颗粒·μL 的浓度范围内对 MCF-7 细胞来源的外泌体做出响应,检测限为 158 个颗粒·μL。此外,该适体传感器已扩展到来自乳腺癌患者的血清样本,并表现出优异的特异性。总之,该适体传感器具有灵敏度高、操作简单、成本低等优点,完全能够在临床上广泛应用于肿瘤监测。