Department of Electrical and Biomedical Engineering, University of Nevada Reno, Reno, NV 89557-0260, USA.
Biosens Bioelectron. 2011 Dec 15;30(1):342-6. doi: 10.1016/j.bios.2011.09.034. Epub 2011 Oct 2.
C-reactive protein (CRP), a 115 kDa pentameric protein, is one of the important cardiac biomarkers that are indicative of coronary heart events. Sensitive detection of CRP in human serum is critical for the diagnosis of coronary heart disease. This work presents a sensitive sandwich immunoassay for the detection of CRP in human serum using zinc sulfide (ZnS) nanoparticles as novel fluorescence signal transducers. In this assay, monoclonal anti-CRP antibodies are used to capture CRP in human serum, and then the captured CRPs are incubated with biotinylated monoclonal anti-CRP and Neutravidin coated ZnS nanoparticle to form sandwich immunocomplexes. Quantification of CRP occurs when zinc ions released from ZnS nanoparticle labels are mixed with zinc-ion sensitive fluorescence indicator Fluozin-3 for fluorescence generation. The developed assay presents a detection limit around 10 pM and a detection range with more than two orders of magnitude.
C-反应蛋白(CRP)是一种 115kDa 的五聚体蛋白,是指示冠心病事件的重要心脏生物标志物之一。在人血清中敏感地检测 CRP 对于冠心病的诊断至关重要。本工作提出了一种使用硫化锌(ZnS)纳米粒子作为新型荧光信号转导物检测人血清中 CRP 的灵敏夹心免疫测定法。在该测定法中,使用单克隆抗 CRP 抗体捕获人血清中的 CRP,然后将捕获的 CRP 与生物素化的单克隆抗 CRP 和链霉亲和素包被的 ZnS 纳米粒子孵育以形成夹心免疫复合物。当从 ZnS 纳米粒子标记物释放的锌离子与锌离子敏感的荧光指示剂 Fluozin-3 混合以产生荧光时,发生 CRP 的定量。所开发的测定法的检测限约为 10pM,检测范围超过两个数量级。