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Nanobody engineering for enhanced-sensitivity rapid COVID-19 tests.

Eunji Jeong1, Seo Yeong Oh2, Seong Uk Son2,3

  • 1Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul 03722, Republic of Korea. haam@yonsei.ac.kr.

Nanoscale
|September 15, 2025
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Summary

This study introduces a novel nanobody-based lateral flow immunoassay (LFIA) for rapid and early detection of COVID-19. The new diagnostic test shows improved sensitivity and specificity compared to traditional antibody-based tests, aiding infectious disease surveillance.

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Area of Science:

  • Immunotechnology
  • Diagnostic Assays
  • Virology

Background:

  • Nanobodies offer advantages over conventional antibodies in diagnostic applications due to their small size, stability, and ease of production.
  • Conventional antibody-based lateral flow immunoassays (LFIAs) have limitations in sensitivity and specificity for detecting infectious diseases like COVID-19.
  • The global impact of coronavirus disease (COVID-19) necessitates improved diagnostic tools for early detection and outbreak control.

Purpose of the Study:

  • To develop and validate a nanobody-based LFIA for the rapid and early detection of SARS-CoV-2.
  • To address the limitations of existing antibody-based LFIAs for COVID-19 diagnosis.
  • To evaluate the sensitivity, specificity, and reliability of the novel nanobody-based LFIA.

Main Methods:

  • Development of nanobodies with high specificity against the SARS-CoV-2 nucleocapsid protein.
  • Conjugation of nanobodies to gold nanoparticles (GNPs) to create nanobody-GNPs (nb-GNPs).
  • Optimization of nb-GNPs for incorporation into the LFIA platform and testing with clinical samples.

Main Results:

  • The nanobody-based LFIA achieved a lower limit of detection (10^3.15 PFU mL^-1) and higher reliability (R^2 = 0.9874) than antibody-based tests.
  • Results were obtained within 15 minutes, demonstrating strong discriminative power for clinical samples.
  • The assay showed high specificity, distinguishing COVID-19 from other respiratory viruses, with excellent sensitivity and accuracy.

Conclusions:

  • The developed nanobody-based LFIA is a sensitive, specific, and rapid diagnostic tool for COVID-19 detection.
  • This nanobody-LFIA serves as a viable alternative to conventional antibody-based LFIAs, offering improved performance.
  • The technology can significantly contribute to SARS-CoV-2 surveillance and future pandemic preparedness.