A homogeneous biosensor for Human Epididymis Protein 4 based on upconversion luminescence resonance energy transfer
A homogeneous biosensor for Human Epididymis Protein 4 based on upconversion luminescence resonance energy transfer
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基于上转换发光共振能量转移的人附睾蛋白4均质生物传感器
DOI：
10.1016/j.microc.2021.106083
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发表时间：
2021-02
期刊：
影响因子：
--
通讯作者：
Chunya Li
中科院分区：
文献类型：
--
作者：
Xiuqing Ma;Guiling Huang;Miantai Ye;Xiaojing Zhang;Yanying Wang;Tao Liang;Hongping Deng;Chunya Li
人附睾蛋白4 （HE4）是一种广泛应用于卵巢癌早期诊断的生物标志物。发光共振能量转移（LRET）是一种简便、特异的均相分析技术，在生物检测中具有显著优势。镧系掺杂上转换纳米粒子（UCNPs）具有近红外（NIR）激发特性，可以有效消除生物样品自身发光的干扰，是构建lret探针的理想材料。在此，基于LRET策略制备了HE4纳米探针，其中UCNPs作为能量供体。选择商业有机染料BHQ-1作为能量受体，其吸收与掺杂Tm3+的UCNPs的蓝色发射有很好的重叠。用含有HE4适体序列的分子信标（MB）的两端标记这两个组分。由于UCNPs和BHQ-1的接近性和光谱重叠，82%的上转换发光（UCL）可以通过LRET猝灭。HE4存在时，发夹结构被打开，HE4与其适体序列发生强相互作用，导致UCNPs和BHQ-1分离，抑制LRET。在0.4 ~ 7.0 ng/mL范围内，UCL强度与HE4浓度的对数呈良好的线性关系，HEPES缓冲液的LOD为0.021 ng/mL。该探针具有良好的选择性和稳定性，可成功应用于人血清中HE4的定量分析。这可能是卵巢癌早期诊断的可靠方法。
Human epididymis protein 4 (HE4) is a widely used biomarker for the early diagnosis of ovarian cancer. Luminescence resonance energy transfer (LRET) is a homogeneous assay technique with great simplicity and specificity, which shows a significant advantage in a bioassay. Lanthanide-doped upconverting nanoparticles (UCNPs) were an ideal material to construct LRET-based probes, because their near-infrared (NIR)-excitable property can effectively eliminate the interference from autoluminescence of biosamples. Herein, a HE4 nanoprobe was fabricated based on LRET strategy, in which UCNPs was employed as an energy donor. The commercial organic dye BHQ-1, whose absorption overlaps well with the blue emission of Tm3+-doped UCNPs, was chosen as an energy acceptor. The two components were tagged with the two ends of a molecular beacon (MB) containing HE4 aptamer sequence. Due to the proximity and spectral overlap of UCNPs and BHQ-1, 82% of upconversion luminescence (UCL) can be quenched through LRET. In presence of HE4, the hairpin structure was opened for the strong interaction between HE4 and its aptamer sequence, which leads to the separation of UCNPs and BHQ-1 and inhibition of LRET. There is a good linear relationship between UCL intensity and the logarithm of HE4 concentration in the range from 0.4 ng/mL to 7.0 ng/mL, with a LOD of 0.021 ng/mL in HEPES buffer. This probe exhibits excellent selectivity and stability, and can also be successfully applied in HE4 quantification in human serum. It might be a reliable method for the early diagnosis of ovarian cancer.
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