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基于 rGO-TEPA-Thi-Au 纳米复合材料和新型 RuPdPt 三元纳米合金颗粒的用于检测 MCP-1 的灵敏三明治型免疫传感器。

A sensitive sandwich-type immunosensor for the detection of MCP-1 based on a rGO-TEPA-Thi-Au nanocomposite and novel RuPdPt trimetallic nanoalloy particles.

机构信息

College of Pharmacy, Institute of Life Science and School of Public Health, Chongqing Medical University, Chongqing 400016, PR China.

College of Pharmacy, Institute of Life Science and School of Public Health, Chongqing Medical University, Chongqing 400016, PR China.

出版信息

Biosens Bioelectron. 2019 Apr 15;131:67-73. doi: 10.1016/j.bios.2019.02.021. Epub 2019 Feb 19.

Abstract

A novel electrochemical immunosensor was proposed for the detection of monocyte chemoattractant protein-1 (MCP-1), a biomarker of cardiovascular disease. Due to thionine (Thi) possessing electroactive redox properties, a one-step approach was utilized to synthesize a reduced graphene oxide-tetraethylene-thionine-Au (rGO-TEPA-Thi-Au) nanocomposite at room temperature using the synergistic effect of Thi and rGO-TEPA towards HAuCl. We obtained the excellent matrix material, which immobilized more primary antibody MCP-1-Ab on rGO-TEPA on a modified glassy carbon electrode (GCE). To further enhance the sensitivity of the sensor, a novel signal generation and amplification strategy was developed for detection. RuPdPt trimetallic nanoalloy particles (RuPdPt TNPs), a novel nanomaterial, were synthesized by a one-pot method, displayed a uniform morphology as well as good electrochemical activity and bound with the secondary antibodies against MCP-1 via the Pt-NH bond. Most importantly, RuPdPt TNPs have a significant ability to catalyze HO to produce an electron. The electrochemical signal was highly amplified because the electrochemical signal was primarily derived from the synergistic catalysis of HO by RuPdPt TNPs and recorded by chronoamperometry. Under the optimal conditions, this newly designed biosensor exhibited sensitive detection of MCP-1 in the range from 20 fg mL to 1000 pg mL, with a detection limit of 8.9 fg mL (based on a S/N = 3). Additionally, the designed immunosensor showed acceptable selectivity, reproducibility and stability. This immunosensor is a promising strategy for analyzing clinical serum samples in the future.

摘要

一种用于检测单核细胞趋化蛋白-1(MCP-1)的新型电化学免疫传感器,MCP-1 是心血管疾病的生物标志物。由于硫堇(Thi)具有电活性氧化还原特性,因此利用 Thi 和 rGO-TEPA 对 HAuCl 的协同作用,在室温下采用一步法合成了还原氧化石墨烯-四乙烯五胺-硫堇-金(rGO-TEPA-Thi-Au)纳米复合材料。我们获得了优异的基质材料,该材料在修饰后的玻碳电极(GCE)上将更多的单核细胞趋化蛋白-1 抗体(MCP-1-Ab)固定在 rGO-TEPA 上。为了进一步提高传感器的灵敏度,开发了一种用于检测的新型信号产生和放大策略。通过一锅法合成了新型纳米材料 RuPdPt 三金属纳米合金颗粒(RuPdPt TNPs),具有均匀的形态以及良好的电化学活性,并通过 Pt-NH 键与针对 MCP-1 的二级抗体结合。最重要的是,RuPdPt TNPs 具有将 HO 催化产生电子的显著能力。由于电化学信号主要源自 RuPdPt TNPs 对 HO 的协同催化作用,并通过计时安培法进行记录,因此电化学信号得到了高度放大。在最佳条件下,该新型生物传感器对 MCP-1 的检测范围从 20 fg mL 到 1000 pg mL,检测限为 8.9 fg mL(基于 S/N=3)。此外,所设计的免疫传感器表现出可接受的选择性、重现性和稳定性。该免疫传感器是未来分析临床血清样本的一种有前途的策略。

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