Shi Zhuxuan, Zhang Tongtong, Zhao Yujiao, Zhou Yucheng, Liu Jiyang
School of Chemistry and Chemical Engineering, Zhejiang Sci-Tech University, Hangzhou, 310018, China.
Department of Obstetrics and Gynecology, Hangzhou First People's Hospital, School of Medicine, Westlake University, Hangzhou, 310006, China.
Biosens Bioelectron. 2025 Nov 15;288:117828. doi: 10.1016/j.bios.2025.117828. Epub 2025 Jul 28.
A dual-signal-quenched sandwich electrochemiluminescence (ECL) immunosensor was developed for highly sensitive detection of tumor biomarkers by integrating a silica nanochannel array film (SNF) and glucose oxidase (GOD)-loaded immuno-nanogold. The SNF modified on the supporting electrode provided dual functional domains including nanochannels and the outer surface. The most commonly used ECL emitter, tris(2,2'-bipyridyl) ruthenium (Ru (bpy)), was enriched in the nanochannels of SNF via electrostatic interaction, significantly amplifying the original ECL signal even at low concentrations of Ru (bpy). The outer surface of SNF enabled covalent immobilization of capture antibody (cAb). Gold nanoparticles co-loaded with GOD and detection antibody (dAb) formed a multienzyme-labeled nanoprobe (GOD/dAb@Au). Upon CA 199 (5 U/mL) binding, the dual-antibody sandwich immunocomplex formed on the electrode interface, decreasing the ECL signal (30.1 % reduction in the initial signal) through increased interfacial resistance and reduced Ru (bpy) diffusion. Additionally, GOD catalyzed the conversion of glucose to produce HO, which acted as a quencher, reducing the ECL signal of Ru (bpy) (16.7 % reduction in the initial signal). This dual-signal-quenching strategy enabled sensitive detection of carbohydrate antigen 19-9 (CA19-9) as a proof-of-concept demonstration with a linear range from 50 μU/mL to 50 U/mL and a low limit of detection (LOD) of 1.1 μU/mL.
通过整合二氧化硅纳米通道阵列膜（SNF）和负载葡萄糖氧化酶（GOD）的免疫纳米金，开发了一种双信号猝灭夹心电化学发光（ECL）免疫传感器，用于高灵敏度检测肿瘤生物标志物。修饰在支撑电极上的SNF提供了包括纳米通道和外表面在内的双功能域。最常用的ECL发光体三（2,2'-联吡啶）钌（Ru（bpy））通过静电相互作用富集在SNF的纳米通道中，即使在低浓度的Ru（bpy）下也能显著放大原始ECL信号。SNF的外表面能够共价固定捕获抗体（cAb）。共负载GOD和检测抗体（dAb）的金纳米颗粒形成了多酶标记纳米探针（GOD/dAb@Au）。当CA 199（5 U/mL）结合时，在电极界面形成双抗体夹心免疫复合物，通过增加界面电阻和减少Ru（bpy）扩散降低ECL信号（初始信号降低30.1%）。此外，GOD催化葡萄糖转化产生HO，其作为猝灭剂，降低Ru（bpy）的ECL信号（初始信号降低16.7%）。这种双信号猝灭策略能够灵敏检测糖类抗原19-9（CA19-9），作为概念验证演示，线性范围为50 μU/mL至50 U/mL，检测下限（LOD）低至1.1 μU/mL。
