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Triggered "Capture-and-Release" Enables a High-Affinity Rebinding Strategy for Sensitivity Enhancement in Lateral

Chapman Ho1,2, Clíona McMahon3, John-Paul Ayrton1,2

  • 1London Centre for Nanotechnology, University College London, 17-19 Gordon Street, Bloomsbury, London WC1H 0AH, United Kingdom.

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|November 26, 2025
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Summary

This study introduces the AmpliFold approach to boost the sensitivity of lateral flow assays (LFAs). By using a capture-and-release method, AmpliFold significantly enhances signal detection for point-of-care diagnostics.

Keywords:
capture-and-releasecleavable linkerslateral flow immunoassaysignal amplificationsite-specific protein/antibody modification

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Area of Science:

  • Biomedical Engineering
  • Assay Development
  • Point-of-Care Diagnostics

Background:

  • Lateral flow assays (LFAs) are widely used point-of-care devices due to their cost-effectiveness, speed, and ease of use.
  • However, the sensitivity of LFAs is often constrained by the kinetics of analyte interactions with assay components.
  • Improving these kinetics is crucial for enhancing detection limits in rapid diagnostic tests.

Purpose of the Study:

  • To develop a novel strategy, termed the AmpliFold approach, to overcome limitations in lateral flow assay (LFA) sensitivity.
  • To reduce the dependence on fast associative kinetics between assay components in LFAs.
  • To demonstrate enhanced signal-to-noise detection through a capture-and-release mechanism.

Main Methods:

  • The AmpliFold approach utilizes a "capture-and-release" strategy with triggered release of analyte-bound complexes.
  • Anti-HER2 Fab fragments with cleavable biotin linkers were employed for triggered release, and their linker length and bioconjugation were optimized.
  • Dual-affinity gold nanoparticles (AuNPs) decorated with anti-HER2 antibodies were used for signal amplification, combined with titration of capture receptor density.

Main Results:

  • The AmpliFold approach achieved up to a 16-fold improvement in LFA sensitivity by overcoming poor capture kinetics at low receptor densities.
  • It successfully addressed issues with large nanoparticle diffusivity and surface binding kinetics in LFAs.
  • A 12-fold sensitivity enhancement was observed using AmpliFold with 150 nm AuNPs in both buffer and human serum samples.

Conclusions:

  • The AmpliFold approach provides a viable strategy for enhancing LFA kinetics and sensitivity using established chemistries.
  • This method offers a rapid (<30 min), equipment-free route to improve diagnostic test performance.
  • The capture-and-release mechanism represents a significant advancement for sensitive and reliable point-of-care diagnostics.