Torrente-Rodríguez Rebeca M, Ruiz-Valdepeñas Montiel Víctor, Campuzano Susana, Pedrero María, Farchado Meryem, Vargas Eva, Manuel de Villena F Javier, Garranzo-Asensio María, Barderas Rodrigo, Pingarrón José M
Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, Madrid, Spain.
Departamento de Bioquímica y Biología Molecular, Facultad de CC. Químicas, Universidad Complutense de Madrid, Madrid, Spain.
PLoS One. 2017 Apr 4;12(4):e0175056. doi: 10.1371/journal.pone.0175056. eCollection 2017.
The first electrochemical immunosensor for the determination of fibroblast growth factor receptor 4 (FGFR4) biomarker is reported in this work. The biosensor involves a sandwich configuration with covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic microcarriers (HOOC-MBs) and amperometric detection at disposable carbon screen-printed electrodes (SPCEs). The biosensor exhibits a great analytical performance regarding selectivity for the target protein and a low LOD of 48.2 pg mL-1. The electrochemical platform was successfully applied for the determination of FGFR4 in different cancer cell lysates without any apparent matrix effect after a simple sample dilution and using only 2.5 μg of the raw lysate. Comparison of the results with those provided by a commercial ELISA kit shows competitive advantages by using the developed immunosensor in terms of simplicity, analysis time, and portability and cost-affordability of the required instrumentation for the accurate determination of FGFR4 in cell lysates.
本工作报道了首个用于测定成纤维细胞生长因子受体4(FGFR4)生物标志物的电化学免疫传感器。该生物传感器采用夹心结构,将特异性捕获抗体共价固定在活化的羧基修饰磁性微载体(HOOC-MBs)上,并在一次性碳丝网印刷电极(SPCEs)上进行安培检测。该生物传感器在对目标蛋白的选择性方面表现出优异的分析性能,检测限低至48.2 pg mL-1。该电化学平台经过简单的样品稀释,仅使用2.5 μg原始裂解液,成功应用于不同癌细胞裂解液中FGFR4的测定,且无明显基质效应。将结果与商用ELISA试剂盒提供的结果进行比较,结果表明,所开发的免疫传感器在简单性、分析时间、便携性以及用于准确测定细胞裂解液中FGFR4所需仪器的成本可承受性方面具有竞争优势。