This work reports the development of the first integrated electrochemical immunosensor for the determination of the ligand receptor activator nuclear factor‐κB (RANKL), a biomarker that plays an important role in the regulation of bone resorption process and, as reported recently, in oncology by modulating the immune system response facilitating the tumor growth and metastatic process during cancer development. The developed platform involves a sandwich‐type immunoassay with covalent immobilization of biotinylated capture antibodies onto streptavidin/4‐aminobenzoic acid ( p ‐ABA) grafted screen‐printed carbon electrodes (bCAb‐Strep/ p ‐ABA‐SPCE) and the use of an hybrid nanomaterial composed of gold nanoparticles (AuNPs) and multi‐walled carbon nanotubes (MWCNTs) as nanocarrier of multiple detector antibody and HRP molecules. Amperometric detection with the H 2 O 2 /hydroquinone (HQ) system allowed the construction of a linear calibration plot for RANKL standards between 10.4 and 1,000 pg mL ‐1 with a LOD of 3.1 pg mL ‐1 . The determination implied simple handling protocols and short assay times. The developed immunosensor was applied to the determination of RANKL in human serum of patients diagnosed with colorectal cancer (CRC) or rheumatoid arthritis (RA). Each determination required only 5 μL of undiluted serum and took less than 2 h counting since the preparation of blocked‐bCAb‐Strep/ p ‐ABA‐SPCE.

中文翻译：

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碳/无机杂化纳米结构作为电化学免疫传感器信号的载体。第一个用于炎症和转移过程的生物传感器，生物标记物RANK-配体

这项工作报告了第一个用于确定配体受体激活物核因子κB（RANKL）的集成电化学免疫传感器的开发，RANKL是一种生物标志物，在骨吸收过程的调节中起着重要作用，最近在肿瘤学中也报道了调节免疫系统反应，促进癌症发展过程中的肿瘤生长和转移过程。开发的平台涉及将生物素化的捕获抗体共价固定在链霉亲和素/ 4-氨基苯甲酸（p-ABA）嫁接的丝网印刷碳电极（bCAb-Strep / p-ABA-SPCE）上的三明治型免疫测定法，以及使用由金纳米颗粒（AuNPs）和多壁碳纳米管（MWCNTs）组成的杂化纳米材料，可作为多种检测抗体和HRP分子的纳米载体。使用H 2 O 2 /对苯二酚（HQ）系统进行安培检测可以构建RANKL标准的线性校正图，其LOD为3.1 pg mL -1且在10.4和1,000 pg mL -1之间。该测定暗示简单的操作规程和较短的测定时间。研发的免疫传感器用于确定患有大肠癌（CRC）或类风湿关节炎（RA）的患者血清中的RANKL。自制备封闭的bCAb-Strep / p-ABA-SPCE以来，每次测定仅需5μL未稀释的血清，计数时间不到2小时。研发的免疫传感器用于确定患有大肠癌（CRC）或类风湿关节炎（RA）的患者血清中的RANKL。自制备封闭的bCAb-Strep / p-ABA-SPCE以来，每次测定仅需5μL未稀释的血清，计数时间不到2小时。研发的免疫传感器用于确定患有大肠癌（CRC）或类风湿关节炎（RA）的患者血清中的RANKL。自制备封闭的bCAb-Strep / p-ABA-SPCE以来，每次测定仅需5μL未稀释的血清，计数时间不到2小时。