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Updated: Jun 26, 2026

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer
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Detecting protein-ligand binding on supported bilayers by local pH modulation.

Hyunsook Jung1, Aaron D Robison, Paul S Cremer

  • 1Department of Chemistry, Texas A&M University, P.O. Box 30012, College Station, Texas 77843-3012, USA.

Journal of the American Chemical Society
|January 8, 2009
PubMed
Summary

This study introduces a sensitive method to detect protein-ligand binding by measuring pH changes at interfaces. The technique accurately quantizes binding affinities and offers a low detection limit for biomolecular interactions.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Materials Science

Background:

  • Detecting protein-ligand interactions at interfaces is crucial for understanding biological processes.
  • Existing methods may lack sensitivity or require complex labeling.
  • A novel, label-free approach is needed for sensitive interfacial binding detection.

Purpose of the Study:

  • To develop a highly sensitive technique for detecting protein-ligand binding at the liquid/solid interface.
  • To utilize pH modulation at the interface upon protein binding as a detection signal.
  • To quantify binding affinities and establish the limit of detection for this new method.

Main Methods:

  • Incorporation of a pH-sensitive dye (ortho-Texas Red DHPE) into supported phospholipid bilayers.

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  • Detection of interfacial pH changes caused by protein binding events.
  • Utilizing fluorescence modulation of the dye to quantify binding and determine equilibrium dissociation constants (K(D)).

    • Main Results:

    • The method successfully detected antibiotin/biotin and cholera toxin B subunit/GM(1) binding.
    • Equilibrium dissociation constants were accurately determined, showing excellent agreement with established techniques.
    • A very low limit of detection (approximately 350 fM) was achieved, comparable to or exceeding surface plasmon resonance.

    Conclusions:

    • The developed technique offers a highly sensitive and accurate method for studying protein-ligand interactions at interfaces.
    • The pH-modulation detection strategy provides a label-free and robust assay.
    • The assay's sensitivity and potential for multiplexed measurements (imaging mode) highlight its broad applicability in biological and materials science.