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Drug Target Engagement Using Coupled Cellular Thermal Shift Assay-Acoustic Reverse-Phase Protein Array.
Adrien Herledan1, Marine Andres1,2, Aurore Lejeune-Dodge3
1University of Lille, Inserm, Institut Pasteur de Lille, U1177-Drugs and Molecules for Living Systems, Lille, France.
SLAS Discovery : Advancing Life Sciences R & D
|December 31, 2019
Summary
A new assay combines nanoacoustics and protein arrays for faster, cost-effective drug target engagement analysis. This cellular thermal shift assay (CETSA) method requires fewer cells and resources, accelerating drug discovery.
Area of Science:
- Biochemistry
- Pharmacology
- Biotechnology
Background:
- Cellular thermal shift assay (CETSA) measures drug-target engagement by detecting ligand-induced protein thermal stability changes.
- Traditional CETSA analysis methods (mass spectrometry, Western blotting, AlphaScreen) are often time-consuming and require substantial sample amounts.
- Limitations in throughput and protein requirements hinder the widespread application of CETSA in drug discovery.
Purpose of the Study:
- To develop a novel, high-throughput, and resource-efficient method for analyzing drug-target engagement using CETSA.
- To overcome the limitations of existing CETSA detection techniques regarding time, cell usage, and sample quantity.
- To validate a new coupled assay for assessing the engagement of drug candidates with their intended targets in a cellular context.
Main Methods:
- Development of a coupled assay integrating a nanoacoustic transfer system with reverse-phase protein array (RPPA) technology for CETSA.
- Application of the CETSA-acoustic reverse-phase protein array (CETSA-aRPPA) to analyze protein thermal stability changes in response to drug treatment.
- Validation of the CETSA-aRPPA technology using inhibitors targeting insulin-degrading enzyme (IDE), a key enzyme in diabetes and Alzheimer's disease.
Main Results:
- The developed CETSA-aRPPA enables simultaneous analysis of numerous conditions and drug-target interactions.
- The assay significantly reduces the required sample size compared to conventional methods.
- Demonstrated rapid, cost-effective, and biological material-saving analysis of drug-target engagement.
Conclusions:
- CETSA-aRPPA offers a significant advancement in drug discovery by enhancing the efficiency and reducing the resource demands of target engagement studies.
- This novel technology facilitates rapid assessment of drug candidates' interaction with their targets in a biologically relevant manner.
- The CETSA-aRPPA method holds promise for accelerating the identification and optimization of novel therapeutics for diseases like diabetes and Alzheimer's.
