Department of Analytical Chemistry, Faculty of Chemistry, University Complutense of Madrid, 28040 Madrid, Spain.
Biosensors (Basel). 2022 Aug 7;12(8):610. doi: 10.3390/bios12080610.
Serum level of CCL5 chemokine is considered an emerging biomarker for multiple sclerosis (MS). Due to the lack of specific assays for this disease, the development of a point-of-care test for rapid detection of MS could lead to avoiding diagnostics delays. In this paper, we report the first electrochemical immunoplatform for quantification of the CCL5 biomarker at the clinically required levels, able to discriminate between patients diagnosed with MS and healthy individuals. The immunosensing device involves protein capture from biological samples by complexation with biotinylated specific antibodies immobilized onto neutravidin-functionalized microparticles and sandwich assay with anti-CCL5 antibody and IgG labelled with horseradish peroxidase (HRP) for the enzyme-catalyzed amperometric detection of HO using hydroquinone (HQ) as the redox mediator. The method shows excellent analytical performance for clinical application with a wide linear range of concentrations (0.1-300 ng·mL CCL5, R = 0.998) and a low detection limit (40 pg·mL CCL5). The biosensing platform was applied to the determination of the CCL5 endogenous content in 100-fold diluted sera both from healthy individuals and patients diagnosed with MS, with no further sample treatment in just two hours. The results were successfully compared with those obtained by the ELISA methodology.
血清 CCL5 趋化因子水平被认为是多发性硬化症（MS）的一种新兴生物标志物。由于针对这种疾病缺乏特异性检测方法，因此开发用于快速检测 MS 的即时检测（POCT）测试可能会避免诊断延误。在本文中，我们报告了第一个用于在临床所需水平定量 CCL5 生物标志物的电化学免疫平台，该平台能够区分诊断为 MS 的患者和健康个体。免疫传感设备涉及通过与固定在链霉亲和素功能化微颗粒上的生物素化特异性抗体复合物从生物样品中捕获蛋白质，并使用抗 CCL5 抗体和标记有辣根过氧化物酶（HRP）的 IgG 进行夹心测定，用于使用对苯二酚（HQ）作为氧化还原介体进行 HO 的酶催化安培检测。该方法具有出色的分析性能，可用于临床应用，具有广泛的浓度线性范围（0.1-300 ng·mL CCL5，R = 0.998）和低检测限（40 pg·mL CCL5）。该生物传感平台应用于测定经过 100 倍稀释的血清中 CCL5 的内源性含量，来自健康个体和诊断为 MS 的患者的血清均无需进一步处理，仅需两个小时即可完成。结果与 ELISA 方法学获得的结果成功进行了比较。
