Zhang Liding, Su Ying, Liang Xiaohan, Cao Kai, Luo Qingming, Luo Haiming
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology (HUST), Wuhan, 430074 China.
MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan, 430074 China.
Nano Res. 2023;16(5):7459-7469. doi: 10.1007/s12274-022-5354-4. Epub 2023 Feb 28.
Phosphorylation of tau at Ser (396, 404) (p-tau) is one of the earliest phosphorylation events, and plasma p-tau level appears to be a potentially promising biomarker of Alzheimer's disease (AD). The low abundance and easy degradation of p-tau in the plasma make the lateral flow assay (LFA) a suitable choice for point-of-care detection of plasma p-tau levels. Herein, based on our screening of a pair of p-tau-specific antibodies, we developed a colorimetric and surface-enhanced Raman scattering (SERS) dual-readout LFA for the rapid, highly sensitive, and robust detection of plasma p-tau levels. This LFA realized a detection limit of 60 pg/mL by the naked eye or 3.8 pg/mL by SERS without cross-reacting with other tau species. More importantly, LFA rapidly and accurately differentiated AD patients from healthy controls, suggesting that it has the potential for clinical point-of-care application in AD diagnosis. This dual-readout LFA has the advantages of simple operation, rapid, and ultra-sensitive detection, providing a new way for early AD diagnosis and intervention, especially in primary and community AD screening.
Supplementary material (characterization of AuNPs and 4-MBA@AuNP probe; the optimal 4-MBA load for AuNPs; the optimal K2CO3 volumes for 4-MBA@AuNP-3G5 conjugates; the optimal 3G5 load for 4-MBA@AuNP conjugates; effect of NaCl concentration on 4-MBA@AuNP-3G5 stability; the linear curve of T-line color and SERS intensity versus different p-tau396,404 concentrations; the comparison of colorimetric-based LFA test results and the diagnosis results; Raman intensities and antibody activity of 4-MBA@AuNP-3G5 before and after storage; colorimetric intensity of dual-readout LFA detecting different concentrations of p-tau396,404 protein; sequence of synthesized peptides used in this study; information of the participants in this study; the information of antibodies used in this study) is available in the online version of this article at 10.1007/s12274-022-5354-4.
tau蛋白在丝氨酸（396、404）位点的磷酸化（p-tau）是最早的磷酸化事件之一，血浆p-tau水平似乎是阿尔茨海默病（AD）一个潜在的有前景的生物标志物。血浆中p-tau丰度低且易降解，使得侧向流动分析（LFA）成为即时检测血浆p-tau水平的合适选择。在此，基于我们对一对p-tau特异性抗体的筛选，我们开发了一种比色和表面增强拉曼散射（SERS）双读出LFA，用于快速、高灵敏度且稳健地检测血浆p-tau水平。这种LFA通过肉眼实现的检测限为60 pg/mL，通过SERS实现的检测限为3.8 pg/mL，且不与其他tau蛋白种类发生交叉反应。更重要的是，LFA能快速准确地区分AD患者和健康对照，表明其在AD诊断的临床即时检测应用中具有潜力。这种双读出LFA具有操作简单、检测快速且超灵敏的优点，为AD的早期诊断和干预提供了一种新方法，尤其适用于基层和社区的AD筛查。
补充材料（金纳米颗粒和4-巯基苯甲酸@金纳米颗粒探针的表征；金纳米颗粒的最佳4-巯基苯甲酸负载量；4-巯基苯甲酸@金纳米颗粒-3G5缀合物的最佳碳酸钾体积；4-巯基苯甲酸@金纳米颗粒缀合物的最佳3G5负载量；氯化钠浓度对4-巯基苯甲酸@金纳米颗粒-3G5稳定性的影响；T线颜色和SERS强度与不同p-tau396,404浓度的线性曲线；基于比色的LFA测试结果与诊断结果的比较；储存前后4-巯基苯甲酸@金纳米颗粒-3G5的拉曼强度和抗体活性；双读出LFA检测不同浓度p-tau396,404蛋白的比色强度；本研究中合成肽的序列；本研究参与者的信息；本研究中使用抗体的信息）可在本文的在线版本中获取，链接为10.1007/s12274-022-5354-4。
