Centre for Biosensors, Bioelectronics and Biodevices (C3Bio) and Department of Electronic and Electrical Engineering, University of Bath, Claverton Down, Bath BA2 7AY, United Kingdom.
Biosens Bioelectron. 2018 Oct 15;117:620-627. doi: 10.1016/j.bios.2018.07.003. Epub 2018 Jul 3.
A novel fluidic-based electrochemical ELISA platform is descried for estimation of the bladder cancer protein markers nuclear mitotic apparatus protein 1 (NUMA1) and complement factor H-related 1 (CFHR1). The platform uses an off-site chamber for a sandwich immunoassay and performs the electrochemistry on-chip in a separate chamber. The off-site matrices were connected to the sensor chip in a manner that the sensors were exposed only to the final electroactive product for signal detection, thus avoiding interference from other molecules present in the sample. Two off-site matrices using 3D polymethyl methacrylate (PMMA) sheets and 2D polycarbonate (PC) membranes modified with the desired antibodies were investigated. Antibodies for NUMA1 and CFHR1 were utilized for the immunoassay and hair comb structured gold electrodes were used for sensing. Results in 10% synthetic urine reveal that the system can detect NUMA1 and CFHR1 in the 1-100 ng/ml range with high sensitivities of 260 nA/(ng/ml) and 310 nA/(ng/ml), for NUMA1 and CFHR1, respectively; negligible interference from the diluted urine and other molecules has been observed. A fully automated fluidic prototype has also been developed to demonstrate that automation of the process and multiplexing of detection can be achieved in a small footprint benchtop device. The use of off-site matrix-based platforms paves the way towards a new generation of electrochemical immunosensors for biomarker estimation with negligible non-specific interactions and false signals in complex samples.
描述了一种基于流体制备的电化学酶联免疫吸附分析平台，用于膀胱癌蛋白标志物核有丝分裂器蛋白 1（NUMA1）和补体因子 H 相关蛋白 1（CFHR1）的定量分析。该平台采用场外腔室进行三明治免疫分析，并在单独的腔室内进行电化学检测。场外基质通过一种方式与传感器芯片相连，即传感器仅暴露于最终的电活性产物以进行信号检测，从而避免了样品中其他分子的干扰。研究了两种使用三维聚甲基丙烯酸甲酯（PMMA）片和二维聚碳酸酯（PC）膜的场外基质，其中 PMMA 和 PC 膜经过了所需抗体的修饰。用于免疫分析的抗体为 NUMA1 和 CFHR1，而梳状金电极则用于传感。在 10%的人工尿液中的结果表明，该系统能够在 1-100ng/ml 范围内检测到 NUMA1 和 CFHR1，NUMA1 和 CFHR1 的灵敏度分别为 260nA/(ng/ml)和 310nA/(ng/ml)；稀释尿液和其他分子几乎没有干扰。还开发了一种全自动流体原型，以证明可以在小型台式设备中实现该过程的自动化和检测的多重化。场外基质平台的使用为新一代电化学免疫传感器用于生物标志物的定量分析铺平了道路，这些传感器在复杂样品中具有可忽略的非特异性相互作用和假信号。
