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Open AccessArticle

Simultaneous Quantification of Protein Binding Kinetics in Whole Cells with Surface Plasmon Resonance Imaging and Edge Deformation Tracking

by 1,†, 1,2,†, 1,3, 1,* and 1,*
1
Biodesign Center for Bioelectronics and Biosensors, Arizona State University, Tempe, AZ 85287, USA
2
School of Molecular Sciences, Arizona State University, Tempe, AZ 85287, USA
3
School of Electrical, Energy and Computer Engineering, Arizona State University, Tempe, AZ 85287, USA
*
Authors to whom correspondence should be addressed.
These authors contributed equally.
Membranes 2020, 10(9), 247; https://doi.org/10.3390/membranes10090247
Received: 27 August 2020 / Revised: 18 September 2020 / Accepted: 20 September 2020 / Published: 22 September 2020
(This article belongs to the Special Issue Protein-Lipid Interactions as Key Regulators of Cell Function)
Most drugs work by binding to receptors on the cell surface. Quantification of binding kinetics between drug and membrane protein is an essential step in drug discovery. Current methods for measuring binding kinetics involve extracting the membrane protein and labeling, and both have issues. Surface plasmon resonance (SPR) imaging has been demonstrated for quantification of protein binding to cells with single-cell resolution, but it only senses the bottom of the cell and the signal diminishes with the molecule size. We have discovered that ligand binding to the cell surface is accompanied by a small cell membrane deformation, which can be used to measure the binding kinetics by tracking the cell edge deformation. Here, we report the first integration of SPR imaging and cell edge tracking methods in a single device, and we use lectin interaction as a model system to demonstrate the capability of the device. The integration enables the simultaneous collection of complementary information provided by both methods. Edge tracking provides the advantage of small molecule binding detection capability, while the SPR signal scales with the ligand mass and can quantify membrane protein density. The kinetic constants from the two methods were cross-validated and found to be in agreement at the single-cell level. The variation of observed rate constant between the two methods is about 0.009 s−1, which is about the same level as the cell-to-cell variations. This result confirms that both methods can be used to measure whole-cell binding kinetics, and the integration improves the reliability and capability of the measurement. View Full-Text
Keywords: binding kinetics; surface plasmonic resonance; cell edge tracking; whole cell; wheat germ agglutinin binding kinetics; surface plasmonic resonance; cell edge tracking; whole cell; wheat germ agglutinin
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MDPI and ACS Style

Jing, W.; Hunt, A.; Tao, N.; Zhang, F.; Wang, S. Simultaneous Quantification of Protein Binding Kinetics in Whole Cells with Surface Plasmon Resonance Imaging and Edge Deformation Tracking. Membranes 2020, 10, 247.

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