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Streamlining cellular thermal shift assay for ultra-high throughput screening.

Dr Pascal Lambertz1, Loretta Hamacher1, Dr Jana Flegel1

  • 1Bayer, AG, Germany.

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Summary
This summary is machine-generated.

We optimized the Cellular Thermal Shift Assay (CETSA) for ultra-high throughput screening (uHTS) using isothermal methods and a Gradient Peltier Device (GPD). This enhances drug discovery by improving the identification of true drug-target interactions in live cells.

Keywords:
Androgen receptorAutomationCETSAHigh throughput screeningHit Identification

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Area of Science:

  • Biochemistry
  • Pharmacology
  • Drug Discovery

Background:

  • Cellular Thermal Shift Assay (CETSA) is crucial for drug-target interaction studies.
  • Existing CETSA methods face limitations in ultra-high throughput screening (uHTS).

Purpose of the Study:

  • To develop and validate advanced CETSA methodologies for uHTS.
  • To enhance the efficiency and accuracy of drug-target interaction screening in live cells.

Main Methods:

  • Developed an innovative isothermal CETSA platform for 1536-well plate screening.
  • Utilized a Gradient Peltier Device (GPD) for full melting curve CETSA retesting.
  • Employed luminescence detection and controlled thermal ramp-up for adherent cells.

Main Results:

  • Demonstrated strong correlation between optimized CETSA methods and fluorescence polarization assays.
  • Showcased the potential for identifying true binders and minimizing false positives.
  • Validated the utility of the new CETSA platform for high throughput drug discovery.

Conclusions:

  • The optimized isothermal CETSA and GPD-based CETSA are effective for uHTS.
  • These advancements facilitate more efficient screening of live-cell drug-target interactions.
  • The developed platform holds significant promise for accelerating drug discovery pipelines.