An efficient NIR-to-NIR signal-based LRET system for homogeneous competitive immunoassay
An efficient NIR-to-NIR signal-based LRET system for homogeneous competitive immunoassay
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DOI：
10.1016/j.bios.2019.111921
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发表时间：
2020-02-15
影响因子：
12.6
通讯作者：
Lee, Joonseok
中科院分区：
文献类型：
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作者：
Kang, Dongkyu;Lee, Seok;Lee, Joonseok
上转换纳米粒子（UCNP）是基于发光共振能量转移（LRET）的生物应用的有前途的材料。与量子点、金纳米粒子等经典 RET 供体相比，UCNP 在 NIR 照射下可以发射近红外 (NIR)，由于在被称为诊断窗口的 NIR 区域中具有强穿透力和低自发荧光，因此可提供增强的信噪比。在这里，我们报告了第一个有效的基于近红外到近红外信号的 LRET 系统，用于通过同质竞争免疫测定检测黄体酮，选择作为概念验证目标。为了提高LRET的效率，我们构建了惰性核/活性壳/惰性超薄壳UCNPs（NaYF4@NaYF4:Yb,Tm@NaYF4）作为LRET供体，并构建了紧凑的黄体酮/辣根过氧化物酶/IRdyeQC-1（P-HRP-染料）复合物作为LRET受体。与传统供体和受体（分别为 70% 和 50%）相比，设计的供体和受体显示出显着提高的 LRET 效率（供体和受体分别为 95% 和 85%）。使用开发的 NIR-to-NIR LRET 系统，成功检测到黄体酮，具有无背景信号和低检测限（十倍稀释的人血清中为 1.36 pg/ml）。我们相信，这里开发的基于近红外到近红外信号的高效 LRET 系统有潜力作为同质竞争免疫测定的简单探针，并能够快速检测生物标志物。
Upconversion nanoparticles (UCNPs) are promising materials for biological applications based on luminescence resonance energy transfer (LRET). In contrast to classical RET donors such as quantum dots, gold nanoparticles, UCNPs can emit near-infrared (NIR) upon the NIR irradiation, which provides enhanced signal-to-noise due to strong penetration and low autofluorescence in the NIR region known as the diagnostic window. Here we report the first efficient NIR-to-NIR signal-based LRET system for the detection of progesterone, chosen as a proof-ofconcept target, via homogeneous competitive immunoassay. To enhance the efficiency of LRET, we constructed inert-core/active-shell/inert-ultrathin shell UCNPs (NaYF4@NaYF4:Yb,Tm@NaYF4) as an LRET donor and a compact progesterone/horseradish peroxidase/IRdyeQC-1 (P-HRP-dyes) complex as an LRET acceptor. The designed donor and acceptor showed significantly improved LRET efficiencies (95% and 85% for donor and acceptor, respectively) compared with conventional donor and acceptor (70% and 50%, respectively). Using the developed NIR-to-NIR LRET system, progesterone was successfully detected with a background-free signal and low limit of detection (1.36 pg/ml in ten-fold diluted human serum). We believe that the efficient NIR-to-NIR signal-based LRET system developed here has potential as a simple probe for homogeneous competitive immunoassay, with the ability to rapidly detect biomarkers.