Liu Qianshan, Ouyang Yulin, Wang Yunjiao, Zhou Shuo, Zhan Yang, Wang Liang
Chongqing Institute of Green and Intelligent Technology, Chinese Academy of Sciences, Chongqing School, The University of Chinese Academy of Sciences (UCAS Chongqing), Chongqing, 400714, China.
Brain Cognition and Brain Disease Institute, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
Adv Healthc Mater. 2025 Mar 24:e2405058. doi: 10.1002/adhm.202405058.
Due to matrix interference, detection methods for protein biomarkers in body fluids are limited. Commonly used methods often require antibody modification or fluorescent labeling. Furthermore, subtle differences in protein sequences make it more challenging to detect and differentiate multiple biomarkers. This study introduces a novel nanopore-based method for simultaneous, label-free detection of key Alzheimer's disease (AD) biomarkers in biological samples. The technique enables distinguishable and ultrasensitive detection of amyloid-beta peptides (Aβ,Aβ) amyloid precursor protein (APP), and tubulin associated unit (Tau) proteins in cerebrospinal fluid and serum. The method successfully identifies AD biomarkers by directly detecting Aβ in cerebrospinal fluid and can detect age-dependent changes in Aβ levels in AD mice models, demonstrating reliability comparable to established enzyme linked immunosorbent assay (ELISA) assays and brain plaque-staining confocal imaging. Notably, this method achieves significant advancements in detecting 2.1 pm Aβ and 1.5 pm APP, as well as 627 fm Aβ in serum. This improvement in nanopore technology addresses the challenges of detecting Aβ and Tau alterations in complex biological samples and differentiating between similar protein sequences. The study marks a significant advancement in the analysis of pathogenic proteins in physiological samples, also offering a powerful tool for AD research and diagnostics.
由于基质干扰，体液中蛋白质生物标志物的检测方法受到限制。常用方法通常需要抗体修饰或荧光标记。此外，蛋白质序列的细微差异使得检测和区分多种生物标志物更具挑战性。本研究介绍了一种基于新型纳米孔的方法，用于同时无标记检测生物样品中关键的阿尔茨海默病（AD）生物标志物。该技术能够对脑脊液和血清中的β淀粉样肽（Aβ）、淀粉样前体蛋白（APP）和微管相关单位（Tau）蛋白进行可区分的超灵敏检测。该方法通过直接检测脑脊液中的Aβ成功识别AD生物标志物，并能检测AD小鼠模型中Aβ水平的年龄依赖性变化，证明其可靠性与既定的酶联免疫吸附测定（ELISA）和脑斑块染色共聚焦成像相当。值得注意的是，该方法在检测血清中2.1皮摩尔的Aβ、1.5皮摩尔的APP以及627飞摩尔的Aβ方面取得了显著进展。纳米孔技术的这一改进解决了在复杂生物样品中检测Aβ和Tau变化以及区分相似蛋白质序列的挑战。该研究标志着生理样品中致病蛋白质分析的重大进展，也为AD研究和诊断提供了一个强大的工具。
