Torrente-Rodríguez Rebeca M, Campuzano Susana, Ruiz-Valdepeñas-Montiel Víctor, Pedrero María, Fernández-Aceñero M Jesús, Barderas Rodrigo, Pingarrón José M
Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain.
Department of Pathology, Hospital Clínico San Carlos, 28040 Madrid, Spain.
J Pharm Biomed Anal. 2016 Sep 10;129:288-293. doi: 10.1016/j.jpba.2016.07.020. Epub 2016 Jul 15.
A sensitive and rapid method for the determination of the clinically relevant biomarker human endoglin (CD105) in serum samples is presented, involving a magneto-actuated immunoassay and amperometric detection at disposable screen-printed carbon electrodes (SPCEs). Micro-sized magnetic particles were modified with a specific antibody to selectively capture the target protein which was further sandwiched with a secondary HRP-labeled antibody. The immunocomplexes attached to the magnetic carriers were amperometrically detected at SPCEs using the hydroquinone (HQ)/H2O2/HRP system. The magneto-actuated immunosensing platform was able to detect 5 pmoles of endoglin (in 25μL of sample, 0.2μM) in 30min providing statistically similar results to those obtained using a commercial ELISA kit for the determination of endogenous content of endoglin in human serum samples.
本文介绍了一种灵敏且快速的方法，用于测定血清样本中具有临床相关性的生物标志物人内皮糖蛋白（CD105），该方法涉及磁驱动免疫分析以及在一次性丝网印刷碳电极（SPCE）上的安培检测。用特异性抗体修饰微米级磁性颗粒，以选择性捕获目标蛋白，该目标蛋白再与辣根过氧化物酶（HRP）标记的二抗形成夹心结构。使用对苯二酚（HQ）/H₂O₂/HRP体系，在SPCE上对附着于磁性载体的免疫复合物进行安培检测。该磁驱动免疫传感平台能够在30分钟内检测出5皮摩尔的内皮糖蛋白（在25μL样本中，浓度为0.2μM），所得结果与使用商业ELISA试剂盒测定人血清样本中内皮糖蛋白内源性含量的结果在统计学上相似。
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