基于磁性 RuL@SiO2-Au~RuL-Ab2 三明治型纳米免疫复合物的可再生和超灵敏电致化学发光免疫传感器。
A renewable and ultrasensitive electrochemiluminescence immunosenor based on magnetic RuL@SiO2-Au~RuL-Ab2 sandwich-type nano-immunocomplexes.
机构信息
The State key Laboratory of Novel Functional Materials and Preparation Science, Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China.
出版信息
Sensors (Basel). 2011;11(8):7749-62. doi: 10.3390/s110807749. Epub 2011 Aug 5.
An ultrasensitive and renewable electrochemiluminescence (ECL) immunosensor was developed for the detection of tumor markers by combining a newly designed trace tag and streptavidin-coated magnetic particles (SCMPs). The trace tag (RuL@SiO(2)-AuRuL-Ab2) was prepared by loading Ru(bpy)(3)(2+)(RuL)-conjuged secondary antibodies (RuL-Ab2) on RuL@SiO(2) (RuL-doped SiO(2)) doped Au (RuL@SiO(2)-Au). To fabricate the immunosensor, SCMPs were mixed with biotinylated AFP primary antibody (Biotin-Ab1), AFP, and RuL@SiO2-AuRuL-Ab2 complexes, then the resulting SCMP/Biotin-Ab1/AFP/RuL@SiO2-AuRuL-Ab2 (SBAR) sandwich-type immunocomplexes were absorbed on screen printed carbon electrode (SPCE) for detection. The immunocomplexes can be easily washed away from the surface of the SPCE when the magnetic field was removed, which made the immunosensor reusable. The present immunosensor showed a wide linear range of 0.05-100 ng mL(-1) for detecting AFP, with a low detection limit of 0.02 ng mL(-1) (defined as S/N = 3). The method takes advantage of three properties of the immunosensor: firstly, the RuL@SiO(2)-AuRuL-Ab2 composite exhibited dual amplification since SiO(2) could load large amount of reporter molecules (RuL) for signal amplification. Gold particles could provide a large active surface to load more reporter molecules (RuL-Ab2). Accordingly, through the ECL response of RuL and tripropylamine (TPA), a strong ECL signal was obtained and an amplification analysis of protein interaction was achieved. Secondly, the sensor is renewable because the sandwich-type immunocomplexes can be readily absorbed or removed on the SPCE's surface in a magnetic field. Thirdly, the SCMP modified probes can perform the rapid separation and purification of signal antibodies in a magnetic field. Thus, the present immunosensor can simultaneously realize separation, enrichment and determination. It showed potential application for the detection of AFP in human sera.
一种超灵敏且可重复使用的电化学发光(ECL)免疫传感器,通过结合新设计的微量标记物和链霉亲和素包被的磁性颗粒(SCMPs),用于检测肿瘤标志物。微量标记物(RuL@SiO2-AuRuL-Ab2)是通过将 Ru(bpy)(3)(2+)(RuL)缀合的二级抗体(RuL-Ab2)负载在 RuL@SiO2(RuL 掺杂的 SiO2)掺杂的 Au(RuL@SiO2-Au)上来制备的。为了制备免疫传感器,将 SCMPs 与生物素化的 AFP 一级抗体(Biotin-Ab1)、AFP 和 RuL@SiO2-AuRuL-Ab2 复合物混合,然后将所得的 SCMP/Biotin-Ab1/AFP/RuL@SiO2-AuRuL-Ab2(SBAR)三明治型免疫复合物吸附在丝网印刷碳电极(SPCE)上进行检测。当磁场被移除时,免疫复合物可以很容易地从 SPCE 表面被洗掉,这使得免疫传感器可以重复使用。该免疫传感器在检测 AFP 时具有 0.05-100ng mL(-1) 的宽线性范围,检测限低至 0.02ng mL(-1)(定义为 S/N = 3)。该方法利用了免疫传感器的三个特性:首先,由于 SiO2 可以负载大量的报告分子(RuL)进行信号放大,因此 RuL@SiO2-AuRuL-Ab2 复合物表现出双重放大。金颗粒可以提供更大的活性表面来负载更多的报告分子(RuL-Ab2)。因此,通过 RuL 和三丙胺(TPA)的 ECL 响应,获得了强的 ECL 信号,并实现了蛋白质相互作用的放大分析。其次,传感器是可再生的,因为在磁场中,三明治型免疫复合物可以很容易地被吸附或从 SPCE 表面去除。第三,修饰了 SCMP 的探针可以在磁场中快速分离和纯化信号抗体。因此,该免疫传感器可以同时实现分离、富集和测定。它在人血清中 AFP 的检测中显示出了潜在的应用。
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