School of Microelectronics & Tianjin Key Laboratory of Imaging and Sensing Microelectronic Technology, Tianjin University, Tianjin, 300072, P. R. China.
School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, P. R. China.
Talanta. 2022 Jan 1;236:122839. doi: 10.1016/j.talanta.2021.122839. Epub 2021 Sep 3.
Single-cell analysis of proteins is critical to gain precise information regarding the mechanisms that dictate the heterogeneity in cellular phenotypes and their differential response to internal and external stimuli. However, tools that allow sensitive and easy measurement of proteins in individual cells are still limited. The emerging semiconductor-based bioelectronics may provide a new approach to overcome the challenges in this field, however its utility in single-cell protein analysis has not been explored. In this study, we investigated multiple protein detection in single cells by MoS field effect transistors (MoS-FETs) modified with specific biological probes. First, β-actin antibody was connected to the surface of MoS-FETs by covalent bonds, and the fabricated device was tested using β-actin solution with concentrations from 10 to 10 μg/μL. Next, we examined the application of MoS-FET for protein analysis in complex biological samples, and the device showed electrical signal response to human embryonic kidney cell line HEK293T in a dose-dependent manner. Furthermore, we applied this method to analyze individual liver cancer MHCC-97L cells, targeting four cellular proteins, including β-actin, epidermal growth factor receptor, sirtuin-2, and glyceraldehyde-3-phosphate dehydrogenase. The devices modified with corresponding probes could identify the target proteins and showed cell number-dependent responses. As a proof of principle, we demonstrated sensitive and multiplexed detection of proteins in single cells using MoS-FETs. The biosensor and this detection method are cost-efficient and user-friendly with broad application prospects in biological studies and clinical diagnosis.
单细胞分析蛋白质对于获得关于决定细胞表型异质性及其对内部和外部刺激的差异反应的机制的精确信息至关重要。然而，允许在单个细胞中灵敏且易于测量蛋白质的工具仍然有限。新兴的基于半导体的生物电子学可能为克服该领域的挑战提供一种新方法，但其在单细胞蛋白质分析中的应用尚未得到探索。在这项研究中，我们通过用特异性生物探针修饰的 MoS 场效应晶体管 (MoS-FET) 研究了单细胞中的多种蛋白质检测。首先，β-肌动蛋白抗体通过共价键连接到 MoS-FET 的表面，并用浓度从 10 到 10 μg/μL 的β-肌动蛋白溶液测试所制造的器件。接下来，我们研究了 MoS-FET 在复杂生物样品中的蛋白质分析中的应用，该器件以剂量依赖的方式对人胚肾细胞系 HEK293T 表现出电信号响应。此外，我们将该方法应用于分析单个肝癌 MHCC-97L 细胞，针对包括β-肌动蛋白、表皮生长因子受体、sirtuin-2 和甘油醛-3-磷酸脱氢酶在内的四种细胞蛋白。用相应探针修饰的器件可以识别靶蛋白，并表现出与细胞数量相关的响应。作为原理证明，我们使用 MoS-FET 证明了单细胞中蛋白质的灵敏和多重检测。该生物传感器和检测方法具有成本效益且易于使用，在生物研究和临床诊断中有广泛的应用前景。
