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Article

Dissociation Constant of Integrin-RGD Binding in Live Cells from Automated Micropipette and Label-Free Optical Data

1
Centre for Energy Research, Nanobiosensorics Laboratory, Institute of Technical Physics and Materials Science, 1121 Budapest, Hungary
2
Department of Biological Physics, Eötvös Loránd University, 1117 Budapest, Hungary
3
Department of Electronics Technology, Budapest University of Technology and Economics, 1111 Budapest, Hungary
*
Author to whom correspondence should be addressed.
Biosensors 2021, 11(2), 32; https://doi.org/10.3390/bios11020032
Received: 4 January 2021 / Revised: 21 January 2021 / Accepted: 22 January 2021 / Published: 24 January 2021
The binding of integrin proteins to peptide sequences such as arginine-glycine-aspartic acid (RGD) is a crucial step in the adhesion process of mammalian cells. While these bonds can be examined between purified proteins and their ligands, live-cell assays are better suited to gain biologically relevant information. Here we apply a computer-controlled micropipette (CCMP) to measure the dissociation constant (Kd) of integrin-RGD-binding. Surface coatings with varying RGD densities were prepared, and the detachment of single cells from these surfaces was measured by applying a local flow inducing hydrodynamic lifting force on the targeted cells in discrete steps. The average behavior of the populations was then fit according to the chemical law of mass action. To verify the resulting value of Kd2d = (4503 ± 1673) 1/µm2, a resonant waveguide grating based biosensor was used, characterizing and fitting the adhesion kinetics of the cell populations. Both methods yielded a Kd within the same range. Furthermore, an analysis of subpopulations was presented, confirming the ability of CCMP to characterize cell adhesion both on single cell and whole population levels. The introduced methodologies offer convenient and automated routes to quantify the adhesivity of living cells before their further processing. View Full-Text
Keywords: adhesion; micropipette; waveguide; biosensor; two-dimensional dissociation constant; integrin-RGD-binding adhesion; micropipette; waveguide; biosensor; two-dimensional dissociation constant; integrin-RGD-binding
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MDPI and ACS Style

Gerecsei, T.; Chrenkó, P.; Kanyo, N.; Péter, B.; Bonyár, A.; Székács, I.; Szabo, B.; Horvath, R. Dissociation Constant of Integrin-RGD Binding in Live Cells from Automated Micropipette and Label-Free Optical Data. Biosensors 2021, 11, 32. https://doi.org/10.3390/bios11020032

AMA Style

Gerecsei T, Chrenkó P, Kanyo N, Péter B, Bonyár A, Székács I, Szabo B, Horvath R. Dissociation Constant of Integrin-RGD Binding in Live Cells from Automated Micropipette and Label-Free Optical Data. Biosensors. 2021; 11(2):32. https://doi.org/10.3390/bios11020032

Chicago/Turabian Style

Gerecsei, Tamás, Péter Chrenkó, Nicolett Kanyo, Beatrix Péter, Attila Bonyár, Inna Székács, Balint Szabo, and Robert Horvath. 2021. "Dissociation Constant of Integrin-RGD Binding in Live Cells from Automated Micropipette and Label-Free Optical Data" Biosensors 11, no. 2: 32. https://doi.org/10.3390/bios11020032

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