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Biosensors based on potent miniprotein binder for sensitive testing of SARS-CoV‑2 variants of concern

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Abstract

The miniprotein binder TRI2-2 was employed as an antibody alternative to build a single antibody-coupled TRI2-2 based gold nanoparticle-based lateral flow immunoassay (AT–GLFIA) biosensor. The biosensor provides high specificity and affinity binding between TRI2-2 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) spike antigen receptor binding domain (S-RBD). It also enables rapid testing of wild-type (WT), B.1.1.7 (Alpha), B.1.351 (Beta), B.1.617.2 (Delta), P.1 (Gamma), and B.1.1.529 (Omicron) SARS-CoV-2 S-RBD and is at least ~ 16-fold more sensitive than conventional antibody pair-based GLFIA (AP–GLFIA). Besides, we developed a wireless micro-electrochemical assay (WMECA) biosensor based on the TRI2-2, which demonstrates an excellent VOCs testing capability at the pg mL−1 level. Overall, our results demonstrate that integrating miniprotein binders into conventional immunoassay systems is a promising design for improving the testing capabilities of such systems without hard-to-obtain antibody pair, complex reporter design, laborious signal amplification strategies, or specific instrumentation.

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Funding

This work was partially supported by the Shanghai Science and Technology Biomedical Innovation Funds (19441904200) for financial support.

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  1. Liangwen Hao and Xue Li contributed equally to this work.

Authors and Affiliations

  1. Department of Radiology, Tongji Hospital, Shanghai Frontiers Science Center of Nanocatalytic Medicine, The Institute for Biomedical Engineering & Nano Science, School of Medicine, Tongji University, Shanghai, 200065, China

    Liangwen Hao, Xue Li, Hongying Liang, Wenjing Lei, Weitao Yang & Bingbo Zhang

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Hao, L., Li, X., Liang, H. et al. Biosensors based on potent miniprotein binder for sensitive testing of SARS-CoV‑2 variants of concern. Microchim Acta 191, 38 (2024). https://doi.org/10.1007/s00604-023-06113-2

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