Receptor specific adhesion assay for the quantification of integrin–ligand interactions in intact cells using a microplate based, label-free optical biosensor

Inna Szekacs, Norbert Orgovan, Beatrix Peter, Boglarka Kovacs, R. Horváth

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In this proof of principle study, a sensitive, label-free, whole cell approach for determination of the affinity of ligands to cell adhesion receptors is presented. The employed optical biosensor is capable of recording, in real time, the kinetics of cell adhesion with high resolution in a microplate based format. The assay was made receptor specific by measuring cell adhesion on integrin ligand displaying surfaces while tuning the concentration of the integrin ligand under study in the employed cell suspensions. As a case study, Arg-Gly-Asp (RGD) displaying polymer surfaces were used and cell adhesion was quantified by measuring the antagonistic action of echistatin, a disintegrin also containing the RGD motif, on the adhesion of cancer cells. Using this novel methodology the half maximal inhibitory concentration (IC50) for echistatin in living cancer cells was determined to be in the range of 20–40 nM. The introduced methodology is fast, sensitive, and does not require isolated receptors because biological effects are measured in intact, living cells. The utility of this method for study of other type of adhesion receptors and ligands is discussed.

Original languageEnglish
Pages (from-to)729-734
Number of pages6
JournalSensors and Actuators, B: Chemical
Volume256
DOIs
Publication statusPublished - Mar 1 2018

Fingerprint

Cell adhesion
bioinstrumentation
Biosensors
Labels
Assays
adhesion
Adhesion
Ligands
Cells
cells
Integrins
ligands
interactions
Disintegrins
cancer
methodology
Suspensions
Polymers
biological effects
Tuning

Keywords

  • Biosensor
  • Cell adhesion
  • Integrins
  • Ligand-receptor interaction
  • Real-time kinetics

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Instrumentation
  • Condensed Matter Physics
  • Surfaces, Coatings and Films
  • Metals and Alloys
  • Electrical and Electronic Engineering
  • Materials Chemistry

Cite this

Receptor specific adhesion assay for the quantification of integrin–ligand interactions in intact cells using a microplate based, label-free optical biosensor. / Szekacs, Inna; Orgovan, Norbert; Peter, Beatrix; Kovacs, Boglarka; Horváth, R.

In: Sensors and Actuators, B: Chemical, Vol. 256, 01.03.2018, p. 729-734.

Research output: Contribution to journalArticle

@article{0101b4edea484b1ebfb3e3d9b02ebc5e,
title = "Receptor specific adhesion assay for the quantification of integrin–ligand interactions in intact cells using a microplate based, label-free optical biosensor",
abstract = "In this proof of principle study, a sensitive, label-free, whole cell approach for determination of the affinity of ligands to cell adhesion receptors is presented. The employed optical biosensor is capable of recording, in real time, the kinetics of cell adhesion with high resolution in a microplate based format. The assay was made receptor specific by measuring cell adhesion on integrin ligand displaying surfaces while tuning the concentration of the integrin ligand under study in the employed cell suspensions. As a case study, Arg-Gly-Asp (RGD) displaying polymer surfaces were used and cell adhesion was quantified by measuring the antagonistic action of echistatin, a disintegrin also containing the RGD motif, on the adhesion of cancer cells. Using this novel methodology the half maximal inhibitory concentration (IC50) for echistatin in living cancer cells was determined to be in the range of 20–40 nM. The introduced methodology is fast, sensitive, and does not require isolated receptors because biological effects are measured in intact, living cells. The utility of this method for study of other type of adhesion receptors and ligands is discussed.",
keywords = "Biosensor, Cell adhesion, Integrins, Ligand-receptor interaction, Real-time kinetics",
author = "Inna Szekacs and Norbert Orgovan and Beatrix Peter and Boglarka Kovacs and R. Horv{\'a}th",
year = "2018",
month = "3",
day = "1",
doi = "10.1016/j.snb.2017.09.208",
language = "English",
volume = "256",
pages = "729--734",
journal = "Sensors and Actuators, B: Chemical",
issn = "0925-4005",
publisher = "Elsevier",

}

TY - JOUR

T1 - Receptor specific adhesion assay for the quantification of integrin–ligand interactions in intact cells using a microplate based, label-free optical biosensor

AU - Szekacs, Inna

AU - Orgovan, Norbert

AU - Peter, Beatrix

AU - Kovacs, Boglarka

AU - Horváth, R.

PY - 2018/3/1

Y1 - 2018/3/1

N2 - In this proof of principle study, a sensitive, label-free, whole cell approach for determination of the affinity of ligands to cell adhesion receptors is presented. The employed optical biosensor is capable of recording, in real time, the kinetics of cell adhesion with high resolution in a microplate based format. The assay was made receptor specific by measuring cell adhesion on integrin ligand displaying surfaces while tuning the concentration of the integrin ligand under study in the employed cell suspensions. As a case study, Arg-Gly-Asp (RGD) displaying polymer surfaces were used and cell adhesion was quantified by measuring the antagonistic action of echistatin, a disintegrin also containing the RGD motif, on the adhesion of cancer cells. Using this novel methodology the half maximal inhibitory concentration (IC50) for echistatin in living cancer cells was determined to be in the range of 20–40 nM. The introduced methodology is fast, sensitive, and does not require isolated receptors because biological effects are measured in intact, living cells. The utility of this method for study of other type of adhesion receptors and ligands is discussed.

AB - In this proof of principle study, a sensitive, label-free, whole cell approach for determination of the affinity of ligands to cell adhesion receptors is presented. The employed optical biosensor is capable of recording, in real time, the kinetics of cell adhesion with high resolution in a microplate based format. The assay was made receptor specific by measuring cell adhesion on integrin ligand displaying surfaces while tuning the concentration of the integrin ligand under study in the employed cell suspensions. As a case study, Arg-Gly-Asp (RGD) displaying polymer surfaces were used and cell adhesion was quantified by measuring the antagonistic action of echistatin, a disintegrin also containing the RGD motif, on the adhesion of cancer cells. Using this novel methodology the half maximal inhibitory concentration (IC50) for echistatin in living cancer cells was determined to be in the range of 20–40 nM. The introduced methodology is fast, sensitive, and does not require isolated receptors because biological effects are measured in intact, living cells. The utility of this method for study of other type of adhesion receptors and ligands is discussed.

KW - Biosensor

KW - Cell adhesion

KW - Integrins

KW - Ligand-receptor interaction

KW - Real-time kinetics

UR - http://www.scopus.com/inward/record.url?scp=85031503999&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85031503999&partnerID=8YFLogxK

U2 - 10.1016/j.snb.2017.09.208

DO - 10.1016/j.snb.2017.09.208

M3 - Article

AN - SCOPUS:85031503999

VL - 256

SP - 729

EP - 734

JO - Sensors and Actuators, B: Chemical

JF - Sensors and Actuators, B: Chemical

SN - 0925-4005

ER -