State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
Hunan Provincial Clinical Research Center for Metabolic Associated Fatty Liver Disease, Clinical Research Institute, the Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang, Hunan 421002, China.
ACS Sens. 2023 Jul 28;8(7):2583-2590. doi: 10.1021/acssensors.3c00288. Epub 2023 Jun 27.
Exosomes secreted from cells carry rich information from their parent cells, representing a promising biomarker for investigation of diseases. We develop a dual-nanopore biosensor using DNA aptamers to specifically recognize CD63 protein on the exosome's surface, which enables label-free exosome detection based on ionic current change. The sensor allows for sensitive detection of exosomes with a detection limit of 3.4 × 10 particles/mL. The dual-nanopore biosensor was able to form an intrapipette electric circuit for ionic current measurement due to its unique structure, which is crucial to achieve detection of exosome secretion from a single cell. We utilized a microwell array chip to entrap a single cell into a confined microwell with small volume, enabling the accumulation of exosomes with high concentration. The dual-nanopore biosensor was positioned into the microwell with a single cell, and monitoring of exosome secretion from a single cell in different cell lines and under different stimulations has been achieved. Our design may provide a useful platform for developing nanopore biosensors for detecting cell secretions from a single living cell.
细胞分泌的外泌体携带了来自其亲代细胞的丰富信息，是研究疾病的有前途的生物标志物。我们开发了一种使用 DNA 适体特异性识别外泌体表面 CD63 蛋白的双纳米孔生物传感器，该传感器能够基于离子电流变化实现无标记的外泌体检测。该传感器能够以 3.4×10 个颗粒/mL 的检测限实现对外泌体的灵敏检测。由于其独特的结构，双纳米孔生物传感器能够形成管内电回路以进行离子电流测量，这对于实现从单个细胞中检测外泌体的分泌至关重要。我们利用微流控芯片将单个细胞困在小体积的限制微井中，从而能够积累高浓度的外泌体。将双纳米孔生物传感器定位到带有单个细胞的微井中，已经实现了对不同细胞系和不同刺激下单个细胞中外泌体分泌的监测。我们的设计可能为开发用于从单个活细胞中检测细胞分泌的纳米孔生物传感器提供了一个有用的平台。
