Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, P. R. China.
J Mater Chem B. 2020 Apr 29;8(16):3574-3581. doi: 10.1039/c9tb02170a.
In general, protein detection relies primarily on enzyme-linked immunosorbent assays. Here, we constructed a colorimetric and electrochemical dual-mode biosensor for thrombin detection based on the mechanism of aptamer recognition. Magnetic nanobeads (MBs) were used as carriers for separation and enrichment to quickly capture thrombin (TB) in the complex matrix. Also, the combination of MBs and the magnetic electrode array (MEA) effectively avoided the poisoning of the electrode by biological samples. Furthermore, hybridization chain reaction (HCR) was indirectly used to achieve amplification of TB. A large number of horseradish peroxidases (HRPs) were coupled with the amplified long nucleic acid fragments. Based on the color and current response of the substrate TMB catalyzed by HRP, a dual-mode detection system for thrombin was established to ensure the accuracy of the test results. The method had a minimum resolution of 10 nM to the naked eye and an electrochemical detection limit as low as 0.35 nM. In addition, the sensor provided good anti-interference ability in a complex matrix and showed great potential to detect TB in complex samples.
一般来说，蛋白质检测主要依赖于酶联免疫吸附测定法。在这里，我们基于适体识别的机制构建了一种用于凝血酶检测的比色和电化学双模生物传感器。磁性纳米珠 (MBs) 用作分离和富集的载体，以快速捕获复杂基质中的凝血酶 (TB)。此外，MBs 和磁电极阵列 (MEA) 的结合有效地避免了生物样品对电极的毒害。此外，杂交链式反应 (HCR) 被间接用于实现 TB 的放大。大量辣根过氧化物酶 (HRP) 与扩增的长核酸片段结合。基于 HRP 催化底物 TMB 的颜色和电流响应，建立了一种用于凝血酶的双模检测系统，以确保测试结果的准确性。该方法对裸眼的最小分辨率为 10 nM，电化学检测限低至 0.35 nM。此外，该传感器在复杂基质中具有良好的抗干扰能力，在复杂样品中检测 TB 具有很大的潜力。
