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Sensitive and rapid determination of heat shock protein 70 using lateral flow immunostrips and upconversion nanoparticle fluorescence probes
Analyst ( IF 4.2 ) Pub Date : 2022-06-23 , DOI: 10.1039/d2an00742h
Nengying Wu 1, 2 , Yuxi Wei 1 , Lanlan Pan 1 , Xiaolin Yang 1 , Honglan Qi 1 , Qiang Gao 1 , Chengxiao Zhang 1 , Chen-Zhong Li 3
Affiliation  

Heat shock protein 70 (Hsp70), belonging to the heat shock protein (HSP) family, is reported to be a potential diagnostic biomarker. In this work, a lateral flow immunostrip was fabricated for the sensitive and rapid determination of Hsp70 by the incorporation of fluorescence and upconversion nanoparticle probes. The upconversion nanoparticles (UCNPs, size ∼39 nm, λex = 980 nm; λem = 540 nm) consisting of a NaYF4:Yb/Er core and polyacrylic acid-modified shell were covalently coupled with Hsp70 antibodies to form the signal probe, which was characterized by dynamic light scattering and zeta potential analyses. The lateral flow assay (LFA) was constructed based on the sandwich-type immunoassay using a sample pad, a test pad, and an adsorption pad on a PVP backing. Hsp70 antibody, IgG antibody and the signal probe were separately dropped on the test zone, the control zone of the test pad, and the sample pad, respectively. In the sandwich LFA, since two antibodies bind to Hsp70 antigenic epitopes, i.e. specific binding, it provided superior specificity and high sensitivity, making it an ideal sensing platform for complex samples like serum Hsp70 samples. The important parameters for the preparation of the lateral flow immunostrips were optimized. Under the optimized conditions, Hsp70 can be detected using the increased fluorescence intensity of UCNPs with a wide linear range from 0.11 to 12 ng mL−1, low detection limit of 0.06 ng mL−1, small sample volume (120 μL), short assay time (15 min) and good reproducibility. The fluorescence method was successfully applied in the determination of Hsp70 in serum samples with good recovery. By combining the accessibility of the lateral flow immunostrips and upconversion nanoparticles, the fluorescence method can serve as a point-of-care testing method for protein assays with high sensitivity and fast detection.

中文翻译:

使用侧向流动免疫条和上转换纳米颗粒荧光探针灵敏快速地测定热激蛋白 70

据报道,热休克蛋白 70 (Hsp70) 属于热休克蛋白 (HSP) 家族,是一种潜在的诊断生物标志物。在这项工作中,通过掺入荧光和上转换纳米粒子探针,制造了一种用于灵敏和快速测定 Hsp70 的侧流免疫条。上转换纳米粒子(UCNPs,尺寸 ∼39 nm,λ ex = 980 nm;λ em= 540 nm) 由 NaYF4:Yb/Er 核心和聚丙烯酸修饰的外壳组成,与 Hsp70 抗体共价偶联形成信号探针,其特征在于动态光散射和 zeta 电位分析。侧向流动测定法 (LFA) 基于夹心型免疫测定法构建,使用样品垫、测试垫和 PVP 背衬上的吸附垫。Hsp70抗体、IgG抗体和信号探针分别滴在测试区、测试垫的对照区和样品垫上。在夹心 LFA 中,由于两种抗体与 Hsp70 抗原表位结合,特异性结合,它提供了卓越的特异性和高灵敏度,使其成为血清 Hsp70 样品等复杂样品的理想传感平台。优化了制备侧流免疫条的重要参数。在优化的条件下,可以使用 UCNPs 增加的荧光强度检测 Hsp70,其线性范围从 0.11 到 12 ng mL -1,低检测限为 0.06 ng mL -1,样品量小(120 μL),测定时间短(15分钟),重现性好。荧光法成功应用于血清样品中Hsp70的测定,回收率良好。通过结合横向流动免疫条和上转换纳米粒子的可及性,荧光方法可以作为一种高灵敏度和快速检测的蛋白质测定的即时检测方法。
更新日期:2022-06-23
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