Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry , Central China Normal University , Wuhan 430079 , P. R. China.
International Joint Research Center for Intelligent Biosensor Technology and Health , Central China Normal University , Wuhan 430079 , P. R. China.
ACS Sens. 2018 Aug 24;3(8):1553-1561. doi: 10.1021/acssensors.8b00365. Epub 2018 Aug 1.
Cytokines are small proteins and form complicated cytokine networks to report the status of our health. Thus, accurate profiling and sensitive quantification of multiple cytokines is essential to have a comprehensive and accurate understanding of the complex physiological and pathological conditions in the body. In this study, we demonstrated a robust electrochemical immunosensor for the simultaneous detection of three cytokines IL-6, IL-1β, and TNF-α. First, graphene oxides (GO) were loaded with redox probes nile blue (NB), methyl blue (MB), and ferrocene (Fc), followed by covalent attachment of anti-cytokine antibodies for IL-6, IL-1β, and TNF-α, respectively, to obtain Ab-GO-NB, Ab-GO-MB, and Ab-GO-Fc, acting as the signal reporters. The sensing interface was fabricated by attachment of mixed layers of 4-carboxylic phenyl and 4-aminophenyl phosphorylcholine (PPC) to glassy carbon surfaces. After that, the capture monoclonal antibody for IL-6, IL-1β, and TNF-α was modified to the carboxylic acid terminated sensing interface. And finally a sandwich assay was developed. The quantitative detection of three cytokines was achieved by observing the change in electrochemical signal from signal reporters Ab-GO-NB, Ab-GO-MB, and Ab-GO-Fc. The designed system has been successfully used for detection of three cytokines (IL-6, IL-1β, and TNF-α) simultaneously with desirable performance in sensitivity, selectivity, and stability, and recovery of 93.6%-105.5% was achieved for determining cytokines spiked in the whole mouse serum.
细胞因子是小蛋白，它们形成复杂的细胞因子网络来报告我们健康状况。因此，准确分析和敏感定量多种细胞因子对于全面准确地了解体内复杂的生理和病理状况至关重要。在本研究中，我们展示了一种用于同时检测三种细胞因子 IL-6、IL-1β 和 TNF-α 的强大电化学免疫传感器。首先，将氧化石墨烯 (GO) 负载到氧化还原探针尼罗蓝 (NB)、亚甲蓝 (MB) 和二茂铁 (Fc) 上，然后通过共价键将抗细胞因子抗体分别连接到 IL-6、IL-1β 和 TNF-α 上，得到 Ab-GO-NB、Ab-GO-MB 和 Ab-GO-Fc，用作信号报告物。通过将混合层的 4-羧基苯基和 4-氨基苯基膦酸酯 (PPC) 附着到玻璃碳表面来制备传感界面。之后，将用于 IL-6、IL-1β 和 TNF-α 的捕获单克隆抗体修饰到羧酸封端的传感界面上。最后，建立了三明治检测方法。通过观察信号报告物 Ab-GO-NB、Ab-GO-MB 和 Ab-GO-Fc 的电化学信号变化，实现了对三种细胞因子的定量检测。该设计的系统已成功用于同时检测三种细胞因子 (IL-6、IL-1β 和 TNF-α)，具有良好的灵敏度、选择性和稳定性，并且在全鼠血清中测定细胞因子的加标回收率为 93.6%-105.5%。
