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Multiplexible Wash-Free Immunoassay Using Colloidal Assemblies of Magnetic and Photoluminescent Nanoparticles

机译:使用磁性和光致发光纳米粒子的胶体组件的多功转复洗涤免疫测定

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摘要

Colloidal assemblies of nanoparticles possess both the intrinsic and collective properties of their constituent nanoparticles, which are useful in applications where ordinary nanoparticles are not well suited. Here, we report an immunoassay technique based on colloidal nanoparticle assemblies made of iron oxide nanoparticles (magnetic substrate) and manganese-doped zinc sulfide (ZnS:Mn) nano particles (photoluminescent substrate), both of which are functionalized with antibodies to capture target proteins in a sandwich assay format. After magnetic isolation of the iron oxide nanoparticle assemblies and their bound ZnS:Mn nanoparticle assemblies (MZSNAs), photoluminescence of the remaining MZSNAs is measured for the protein quantification, eliminating the need for washing steps and signal amplification. Using human C-reactive protein as a model biomarker, we achieve a detection limit of as low as 0.7 pg/mL, which is more than 1 order of magnitude lower than that of enzyme-linked immunosorbent assay (9.1 pg/mL) performed using the same pair of antibodies, while using only one-tenth of the antibodies. We also confirm the potential for multiplex detection by using two different types of photoluminescent colloidal nanoparticle assemblies simultaneously.
机译:纳米颗粒的胶体组件具有其组成纳米颗粒的内在和集体性质,可用于普通纳米颗粒不太适合的应用中。在这里,我们报告了一种基于氧化铁纳米颗粒(磁性基材)和掺锰锌硫化锌(ZnS:Mn)纳米颗粒(光致发光底物)制成的胶体纳米粒子组件的免疫测定技术,两者都是用抗体捕获靶蛋白的抗体的官能化在三明治测定格式。在氧化铁纳米粒子组件的磁性分离和它们结合的ZnS:Mn纳米颗粒组件(MZSNA)之后,测量剩余的MZSNA的光致发光,用于蛋白质量化,消除了对洗涤步骤和信号放大的需求。使用人C-反应蛋白作为模型生物标志物,我们达到0.7pg / ml的检出限,该检出限为0.7pg / ml,比使用的酶联免疫吸附测定(9.1pg / ml)低1个左右数量级同一对抗体,同时仅使用十分之一的抗体。我们还通过使用两种不同类型的光致发光胶体纳米颗粒组件来确认多重检测的可能性。

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