Label-Free Profiling of Cell Adhesion: Determination of the Dissociation Constant for Native Cell Membrane Adhesion Receptor-Ligand Interaction

Norbert Orgovan, Beatrix Peter, Szilvia Bősze, Jeremy J. Ramsden, Bàlint Szabò, Robert Horvath

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Here we describe the protocol and workflow for a label-free cell adhesion assay utilizing the high-throughput Epic BenchTop (BT) optical biosensor. We also describe how the dissociation constant for the binding between integrins in their native cell membrane and their ligands immobilized on the planar sensor surface can be determined from the biosensor data. To achieve this, cell adhesion has to be measured on surfaces having fine-tuned ligand densities. The present protocol can be applied to determine the dissociation constant of the binding between any matrix adhesion receptor embedded in its native cell membrane and its ligand, provided that a coating molecule with appropriate functionalization is available. The effect of drugs or other chemicals on this molecular interaction and subsequent cellular adhesion can be investigated in a straightforward way.

Original languageEnglish
Pages (from-to)327-338
Number of pages12
JournalMethods in Pharmacology and Toxicology
Volume53
DOIs
Publication statusPublished - Jan 1 2015

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Keywords

  • Adhesion kinetics
  • Adhesion tailoring
  • Cell adhesion
  • Optical biosensor
  • RGD-tuning
  • Resonant waveguide grating

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Pharmacology (medical)

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