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基于磁响应上转换发光共振能量转移(LRET)的生物传感器用于 SARS-CoV-2 刺突蛋白的超高灵敏检测

Magnetic-responsive upconversion luminescence resonance energy transfer (LRET) biosensor for ultrasensitive detection of SARS-CoV-2 spike protein.

机构信息

Department of Biomedical Engineering, The Hong Kong Polytechnic University, 999077, Hong Kong, China.

Department of Applied Physics, The Hong Kong Polytechnic University, 999077, Hong Kong, China.

出版信息

Biosens Bioelectron. 2024 Mar 15;248:115969. doi: 10.1016/j.bios.2023.115969. Epub 2023 Dec 23.

Abstract

Upconversion nanoparticles (UCNPs) are ideal donors for luminescence resonance energy transfer (LRET)-based biosensors due to their excellent upconversion luminescence properties. However, the relatively large size of antibodies and proteins limits the application of UCNPs-based LRET biosensors in protein detection because the large steric hindrance of proteins leads to low energy transfer efficiency between UCNPs and receptors. Herein, we developed a magnetic responsive UCNPs-based LRET biosensor to control the coupling distance between antibody-functionalized UCNPs (Ab-UCNPs) as donors and antibody-PEG linker-magnetic gold nanoparticles (Ab-PEG-MGNs) as acceptors for ultrasensitive and highly selective detection of SARS-CoV-2 spike proteins. Our results showed that this platform reversibly shortened the coupling distance between UCNPs and MGNs and enhanced the LRET signal with a 10-fold increase in the limit of detection (LOD) from 20.6 pg/mL without magnetic modulation to 2.1 pg/mL with magnetic modulation within 1 h. The finite-difference time-domain (FDTD) simulation with cyclic distance change confirmed the distance-dependent LRET efficiency under magnetic modulation, which supported the experimental results. Moreover, the applications of this magnetic-responsive UCNP-based LRET biosensor could be extended to other large-size biomolecule detection.

摘要

上转换纳米粒子(UCNPs)由于其优异的上转换发光性能,是用于基于荧光共振能量转移(LRET)的生物传感器的理想供体。然而,抗体和蛋白质的相对较大尺寸限制了基于 UCNPs 的 LRET 生物传感器在蛋白质检测中的应用,因为蛋白质的大位阻导致 UCNPs 和受体之间的能量转移效率较低。在此,我们开发了一种磁响应的基于 UCNPs 的 LRET 生物传感器,以控制抗体功能化 UCNPs(Ab-UCNPs)作为供体和抗体-PEG 接头-磁性金纳米颗粒(Ab-PEG-MGNs)作为受体之间的耦合距离,用于超灵敏和高度选择性检测 SARS-CoV-2 刺突蛋白。我们的结果表明,该平台可可逆地缩短 UCNPs 和 MGNs 之间的耦合距离,并增强 LRET 信号,检测限(LOD)从无磁场调制时的 20.6 pg/mL 提高了 10 倍,达到磁场调制时的 2.1 pg/mL,仅需 1 小时。具有循环距离变化的有限差分时间域(FDTD)模拟证实了磁场调制下距离依赖性的 LRET 效率,这支持了实验结果。此外,这种磁响应的基于 UCNPs 的 LRET 生物传感器的应用可以扩展到其他大尺寸生物分子的检测。

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