Wang Wei-Bin, Wang Ya-Fei, Li Jian-Jun, Wang Jing-Yuan, Weng Guo-Jun, Zhu Jian, Zhao Jun-Wu
The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi, 710049, China.
The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi, 710049, China; Department of Neurosurgery, Yulin First Hospital, Yulin, Shaanxi, 719000, China.
Biosens Bioelectron. 2025 Jun 15;278:117352. doi: 10.1016/j.bios.2025.117352. Epub 2025 Mar 6.
Herein, we fabricate the graphene oxide-supported nanofilms coated by roughened nanoboxes (GO@AuAgRNB) for the ultrasensitive and simultaneous determination of multiple stroke subtype-specific biomarkers. Initially, Au-Ag roughened nanobox (AuAgRNB) with abundant coupling and tip hotspots is prepared by the partial surface passivation strategy. AuAgRNB is uniformly, densely and firmly assembled onto graphene oxide (GO) by metal-sulfur bonds, generating extensive high-density hotspots. Owing to electromagnetic and chemical enhancement, the surface enhanced Raman scattering (SERS) activity of GO@AuAgRNB is greatly improved with the enhancement factor of 5.78 × 10. Combined with magnetic bead, GO@AuAgRNB was employed to develop a SERS-based immunoassay platform for the simultaneous detection of glial fibrillary acidic protein (GFAP) and retinol binding protein 4 (RBP). The platform demonstrates ultra-sensitivity with detection ranges of 0.1 pg/mL-0.1 μg/mL and limits of detection of 0.16 pg/mL for GFAP and 0.10 pg/mL for RBP. Furthermore, the platform provides superior anti-interference properties, accuracy, and capability for simultaneous detection and practical application. In the detection of clinical patient samples, the receiver operating characteristic curve analysis shows that cut-off values (RBP = 13.51 μg/mL and GFAP = 2.07 ng/mL) can reliably differentiate patients with ischaemic and haemorrhagic stroke. Overall, the ultrasensitive and multiplex immunoassay platform based on GO@AuAgRNB demonstrates high potential in the clinical diagnosis of stroke subtypes.
在此，我们制备了由粗糙纳米盒包覆的氧化石墨烯负载纳米薄膜（GO@AuAgRNB），用于超灵敏同时检测多种中风亚型特异性生物标志物。首先，通过部分表面钝化策略制备具有丰富耦合和尖端热点的金 - 银粗糙纳米盒（AuAgRNB）。AuAgRNB通过金属 - 硫键均匀、密集且牢固地组装在氧化石墨烯（GO）上，产生大量高密度热点。由于电磁和化学增强作用，GO@AuAgRNB的表面增强拉曼散射（SERS）活性大大提高，增强因子为5.78×10。结合磁珠，GO@AuAgRNB被用于开发基于SERS的免疫分析平台，用于同时检测胶质纤维酸性蛋白（GFAP）和视黄醇结合蛋白4（RBP）。该平台具有超灵敏性，检测范围为0.1 pg/mL - 0.1 μg/mL，GFAP的检测限为0.16 pg/mL，RBP的检测限为0.10 pg/mL。此外，该平台具有卓越的抗干扰性能、准确性以及同时检测和实际应用的能力。在临床患者样本检测中，受试者工作特征曲线分析表明，临界值（RBP = 13.51 μg/mL和GFAP = 2.07 ng/mL）能够可靠地区分缺血性和出血性中风患者。总体而言，基于GO@AuAgRNB的超灵敏多重免疫分析平台在中风亚型的临床诊断中显示出巨大潜力。
