Arabnejad Mahdi, Tothill Ibtisam E, Chianella Iva
Faculty of Engineering and Applied Sciences, Cranfield University, Cranfield, Bedfordshire MK43 0AL, UK.
Silveray, Stockport Road West, Stockport SK6 2BP, UK.
Biosensors (Basel). 2024 Dec 18;14(12):624. doi: 10.3390/bios14120624.
Lung cancer is the most common type of cancer diagnosed worldwide and is also among the most fatal. Early detection, before symptoms become evident, is fundamental for patients' survival. Therefore, several lung cancer biomarkers have been proposed to enable a prompt diagnosis, including neuron-specific enolase (NSE) and carcinoembryonic antigen (CEA). NSE and CEA are two serum proteins whose elevated levels have been associated with lung cancer. Hence, in this study, impedimetric biosensors (immunosensors) able to quantify NSE and CEA were developed as proof-of-concept devices for lung cancer diagnosis. The sensing platform exploited for the immunosensors comprises a novel combination of a magnetic platform, screen-printed gold electrode (SPGE), and magnetic nanobeads (MB). The MB were functionalized with antibodies to capture the analyte from the sample and to move it over the sensing area. The immunosensors were then developed by immobilizing another set of antibodies for either CEA or NSE on the SPGE through formation of self-assembled monolayer (SAM). The second set of antibodies enabled a sandwich assay to be formed on the surface of the sensor, while MB manipulation was applied during the sensor performance to depict a microfluidic system and increase antigen-antibody complex formation prior to CEA or NSE detection and quantification. The optimized immunosensors were successfully tested to measure various concentrations of CEA and NSE (0-100 ng/mL) in both phosphate buffer and 100% human serum samples. Clinically relevant detection limits of 0.26 ng/mL and 0.18 ng/mL in buffer and 0.76 ng/mL and 0.52 ng/mL in 100% serum for CEA and NSE, respectively, were achieved via electrochemical impedance spectroscopy with the use of potassium ferri/ferrocyanide as a redox probe. Hence, the two immunosensors demonstrated great potential as tools to be implemented for the early detection of lung cancer.
肺癌是全球诊断出的最常见癌症类型，也是最致命的癌症之一。在症状出现之前进行早期检测是患者生存的关键。因此，人们提出了几种肺癌生物标志物以实现快速诊断，包括神经元特异性烯醇化酶（NSE）和癌胚抗原（CEA）。NSE和CEA是两种血清蛋白，其水平升高与肺癌有关。因此，在本研究中，开发了能够定量NSE和CEA的阻抗生物传感器（免疫传感器），作为肺癌诊断的概念验证设备。用于免疫传感器的传感平台包括磁性平台、丝网印刷金电极（SPGE）和磁性纳米珠（MB）的新型组合。MB用抗体进行功能化，以从样品中捕获分析物并将其移动到传感区域。然后通过自组装单层（SAM）的形成，将另一组针对CEA或NSE的抗体固定在SPGE上，从而开发出免疫传感器。第二组抗体能够在传感器表面形成夹心测定，同时在传感器性能测试期间应用MB操作来描绘微流体系统，并在检测和定量CEA或NSE之前增加抗原-抗体复合物的形成。优化后的免疫传感器成功地用于测量磷酸盐缓冲液和100%人血清样品中不同浓度的CEA和NSE（0-100 ng/mL）。使用铁氰化钾/亚铁氰化钾作为氧化还原探针，通过电化学阻抗谱分别在缓冲液中实现了CEA和NSE的临床相关检测限为0.26 ng/mL和0.18 ng/mL，在100%血清中为0.76 ng/mL和0.52 ng/mL。因此，这两种免疫传感器作为肺癌早期检测工具具有巨大潜力。
