Nanobody-Based Lateral Flow Immunoassay for Rapid Antigen Detection of SARS-CoV-2 and MERS-CoV Proteins

ACS Synth Biol. 2025 Jan 9. doi: 10.1021/acssynbio.4c00592. Online ahead of print.

Abstract

The COVID-19 pandemic has highlighted the critical need for pathogen detection methods that offer both low detection limits and rapid results. Despite advancements in simplifying and enhancing nucleic acid amplification techniques, immunochemical methods remain the preferred methods for mass testing. These methods eliminate the need for specialized laboratories and highly skilled personnel, making home testing feasible. Here, we developed nanobody-based lateral flow assays (LFAs) for the rapid detection of SARS-CoV-2 and MERS-CoV in single and dual formats as point-of-care diagnostic tools. The developed LFAs are highly sensitive and successfully detected analytes at clinically relevant diagnostic cutoff values. Additionally, our results confirmed that the LFAs have a long shelf life and can be produced cost-effectively and with ease.

Keywords: COVID-19; VHH; coronavirus; gold conjugation; lateral flow immunoassay; nanobody.