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首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >In situ amplified photothermal immunoassay for neuron-specific enolase with enhanced sensitivity using Prussian blue nanoparticleloaded liposomes
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In situ amplified photothermal immunoassay for neuron-specific enolase with enhanced sensitivity using Prussian blue nanoparticleloaded liposomes

机译：原位扩增的光热免疫测定，用于神经元特异性烯醇酶，使用普鲁士蓝纳米粉颗粒脂质体具有增强的敏感性

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摘要

Methods based on prussian blue nanoparticles (PBNPs) have been reported for photothermal immunoassays in analytical nanoscience fields but most suffer from low sensitivity and are not beneficial for routine use. Herein, we design an in situ amplified near-infrared (NIR) photothermal immunoassay for the quantitative screening of neuron-specific enolase (NSE) on a portable thermometer using PBNP-encapsulated nanoliposomes as photosensitive materials. Biotinylated liposomes loaded with numerous prussian blue nanoparticles were synthesized through a typical reverse-phase evaporation method. The photothermal immunoassay was carried out in an anti-NSE capture antibody-coated microplate using the biotinylated anti-NSE secondary antibody. With the sandwiched immunoreaction and the biotin-avidin linkage, the subsequent photothermal measurement of PBNPs released from the liposomes with buffered surfactant including Tween 20 was conducted on a digital thermometer under near-infrared 808 nm laser irradiation, accompanied by the convertion of NIR-light wavelength to heat. Under the optimum conditions, the photothermal immunoassay displayed a wide dynamic concentration range of 0.1-100 ng mL~(-1) with a low detection limit for NSE of 0.053 ng mL~(-1). Good reproducibility (RSD ≤ 2.78% for intraassay; RSD ≤ 4.39% for inter-assay), high selectivity against other biomarkers, and a long-term stability (≥94.9% of the initial signal during six-month storage) were acquired in the photothermal immunoassay. Impressively, the analysis of 7 human serum specimens for target NES via the photothermal immunoassay also gave well-matched results with the referenced human NSE enzyme-linked immunosorbent assay.
机译：据报道，基于普鲁士蓝纳米粒子（PBNPS）的方法，用于分析纳米科学领域的光热免疫测定，但大多数患有低敏感性，并且对常规使用无益。在此，我们使用PBNP包封的纳米吡咯作为光敏材料，设计用于在便携式温度计上进行定量筛选神经元特异性烯醇酶（NSE）的原位扩增的近红外（NIR）光热的免疫测定。通过典型的反相蒸发方法合成装载有许多普鲁士蓝纳米颗粒的生物素化脂质体。使用生物素化的抗NSE二抗，在抗NSE捕获抗体涂覆的微孔板中进行光热免疫测定。随着夹层的免疫反应和生物素 - 抗霉素连杆，随后从近红外线808nm激光照射下的数字温度计上进行了从脂质体释放的Pbnps的Pbnps的光热测量，伴随着Nir-Light的转换波长热量。在最佳条件下，光热免疫测定显示出0.1-100ng ml〜（-1）的宽动态浓度范围，NSE的低检测限为0.053 ng ml〜（-1）。良好的再现性（RSD≤2.78％，用于静脉分析; RSD≤4.39％，用于差异≤4.39％，对其他生物标志物的选择性高，并且在六个月内获得了长期稳定性（≥94.9％的初始信号）光热免疫测定。令人印象深刻地，通过光热免疫测定对靶NE的7个人血清样品的分析也使得结果与参考人体NSE酶联免疫吸附测定产生了良好匹配的结果。

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