Development of a Dual-Epitope Nanobody-Based Immunosensor with MXenes@CNTs@AuNPs for Ultrasensitive Detection of RotavirusClick to copy article link

Immunoassays have become essential tools for detecting infectious viruses. However, traditional monoclonal antibody-dependent immunoassays are costly, fragile, and unstable, especially in complex media. To overcome these challenges, we have developed cost-effective, robust, and high-affinity nanobod...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2024-12, Vol.96 (49), p.19678
Main Authors: Hu, Wenjin, Yang, Shixiang, Wang, Xiaolong, Li, Xi, Lei, Liusheng, Lin, Huai, Yuan, Qingbin, Mao, Daqing, Luo, Yi
Format: Article
Language:English
Subjects:
Antigens
Catalase
Complex media
Epitopes
Immobilization
Immunoassay
Immunoassays
Immunosensors
Magnetic probes
Monoclonal antibodies
MXenes
Nanobodies
Robustness
Rotavirus
Sensitivity enhancement
Signal generation
Two dimensional materials
Viruses
Water sampling
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Summary:Immunoassays have become essential tools for detecting infectious viruses. However, traditional monoclonal antibody-dependent immunoassays are costly, fragile, and unstable, especially in complex media. To overcome these challenges, we have developed cost-effective, robust, and high-affinity nanobodies as alternatives to monoclonal antibodies for rapid detection applications. We engineered dual-epitope nanobody (NB) pairs and incorporated them into a sandwich immunosensor design to detect transmitted rotaviruses in rectal swabs and wastewater samples. To further enhance sensitivity, we synthesized an advanced two-dimensional material, MXenes@CNTs@AuNPs, which offers an extensive specific surface area that supports the enrichment and immobilization of NBs. This integration with catalase-modified magnetic probes facilitates signal generation. Subsequently, our sensor achieved a detection limit of 0.0207 pg/mL for the rotavirus VP6 antigen, significantly outperforming commercial antigen kits with a sensitivity enhancement of 3.77 × 105-fold. The exceptional sensor performance extended to specificity, repeatability, stability, and accuracy across various sample types, establishing it as a promising tool for rotavirus detection. This research outlines a viable strategy for creating a robust and ultrasensitive analytical nanoprobe, thereby addressing the critical need for efficient and reliable viral detection methods in various environments.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.4c04826