High sensitivity perovskite-tagged nanobody electrochemiluminescent immunosensor for spike (S1) protein biomarker-based persistent viral reservoir detection

IF 4.9 2区化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioelectrochemistry Pub Date : 2026-03-01 Epub Date: 2025-10-12 DOI:10.1016/j.bioelechem.2025.109137

Jaymi January , Olivier Zwaenepoel , Nelia Sanga , Jan Gettemans , Emmanuel Iwuoha

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引用次数: 0

Abstract

Persistent viral reservoirs (PVR) of SARS-CoV-2 (or long COVID-19) and latent COVID-19 disease have been of great concern to clinicians. Several studies have identified spike protein (S1) as a definitive biomarker for the early detection of persistent viral reservoirs and latent COVID-19. A novel sandwich-format electrochemical immunosensor integrating a nanocomposite material was engineered for rapid and sensitive latent COVID-19 detection. The platform structure, AuSPE||strep|Nb₁|BSA|biotin|S1|strep-LiSmZrO₃-Nb₂ + BSA (AuSPE = gold screen-printed electrode, strep = streptavidin-thiol, Nb₁ = primary nanobody, Nb₂ = secondary nanobody, BSA = bovine serum albumin, and spike (S1) protein = S1), featured a disposable AuSPE modified with strep to anchor a biotinylated camelid Nb₁ specific to the spike protein. A Nb₂ conjugated to streptavidin-labelled LiSmZrO₃ perovskite completed the sandwich complex, enhancing both affinity and signal transduction. Electrochemical responses of the sensor were studied via electrochemiluminescent (ECL) signal transduction. The S1-sensitive sandwich immunosensor had a detection range of 0–1000 pg mL⁻¹ with a limit of detection of 0.04 pg mL⁻¹ via ECL. As feasibility studies, commercial spike protein in buffered solutions and human serum, highlighting the potential for the immunosensor to be used in SARS-CoV-2 patients and PVRs. The nanobody sandwich immunosensor showed excellent stability, selectivity, sensitivity, and reproducibility. The immunosensor serves as a broad PVR for a SARS-CoV-2 screening tool, detecting elevated S1 levels to enable early, targeted diagnostics.

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高灵敏度钙钛矿标记纳米体电化学发光免疫传感器，用于基于刺突（S1）蛋白生物标志物的持久性病毒库检测。

SARS-CoV-2（或长型COVID-19）和潜伏性COVID-19疾病的持续病毒库（PVR）一直是临床医生关注的焦点。一些研究已经确定刺突蛋白（S1）是早期检测持久性病毒库和潜伏性COVID-19的明确生物标志物。设计了一种集成纳米复合材料的新型三明治式电化学免疫传感器，用于快速灵敏地检测COVID-19潜伏性。该平台结构为AuSPE|| |strep|Nb1|BSA|biotin|S1|strep- lismzro3 -Nb2 + BSA (AuSPE =金丝网印刷电极，strep =链亲和素-硫醇，Nb1 =一级纳米体，Nb2 =二级纳米体，BSA =牛血清白蛋白，spike （S1）蛋白= S1)，其特征是用strep修饰的一次性AuSPE来锚定spike蛋白特异性的生物素化的camelid Nb1。Nb2与链霉亲和素标记的LiSmZrO3钙钛矿结合完成了三明治复合物，增强了亲和力和信号转导。通过电化学发光（ECL）信号转导研究了传感器的电化学响应。对s1敏感的夹心免疫传感器检测范围为0 ~ 1000 pg mL-1， ECL检测限为0.04 pg mL-1。作为可行性研究，缓冲溶液和人血清中的商业刺突蛋白，突出了免疫传感器在SARS-CoV-2患者和pvr中的应用潜力。该纳米体夹心免疫传感器具有良好的稳定性、选择性、灵敏度和可重复性。该免疫传感器可作为一种广泛的PVR，用于SARS-CoV-2筛查工具，检测S1水平升高，从而实现早期、有针对性的诊断。

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来源期刊

Bioelectrochemistry 生物-电化学

CiteScore

9.10

自引率

6.00%

发文量

238

审稿时长

38 days

期刊介绍： An International Journal Devoted to Electrochemical Aspects of Biology and Biological Aspects of Electrochemistry Bioelectrochemistry is an international journal devoted to electrochemical principles in biology and biological aspects of electrochemistry. It publishes experimental and theoretical papers dealing with the electrochemical aspects of: • Electrified interfaces (electric double layers, adsorption, electron transfer, protein electrochemistry, basic principles of biosensors, biosensor interfaces and bio-nanosensor design and construction. • Electric and magnetic field effects (field-dependent processes, field interactions with molecules, intramolecular field effects, sensory systems for electric and magnetic fields, molecular and cellular mechanisms) • Bioenergetics and signal transduction (energy conversion, photosynthetic and visual membranes) • Biomembranes and model membranes (thermodynamics and mechanics, membrane transport, electroporation, fusion and insertion) • Electrochemical applications in medicine and biotechnology (drug delivery and gene transfer to cells and tissues, iontophoresis, skin electroporation, injury and repair). • Organization and use of arrays in-vitro and in-vivo, including as part of feedback control. • Electrochemical interrogation of biofilms as generated by microorganisms and tissue reaction associated with medical implants.