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Using High Content Imaging to Quantify Target Engagement in Adherent Cells.

Hanna Axelsson1, Helena Almqvist1, Brinton Seashore-Ludlow2

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This study presents an advanced cellular thermal shift assay (CETSA) for high-throughput drug target engagement analysis. The modified method quantifies small molecule-protein interactions in living cells without target modification, crucial for drug development.

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Area of Science:

  • Biochemistry
  • Pharmacology
  • Cell Biology

Background:

  • Quantifying small molecule-protein interactions is vital for drug discovery and validation.
  • Existing methods often require target or molecule modification, posing technical challenges.
  • Cellular thermal shift assay (CETSA) monitors target engagement in live cells.

Purpose of the Study:

  • To adapt the CETSA protocol for high-throughput measurements.
  • To enable target engagement analysis at the single-cell level with subcellular localization.
  • To facilitate in-depth characterization of compound-target interactions in diverse cell populations.

Main Methods:

  • Adaptation of the original CETSA protocol.
  • Implementation of high-throughput measurement capabilities.
  • Preservation of subcellular localization data at the single-cell level.

Main Results:

  • A modified CETSA protocol enabling high-throughput analysis.
  • Successful retention of subcellular localization information.
  • Demonstrated utility for studying compound-target interactions in heterogeneous cell populations.

Conclusions:

  • The adapted CETSA protocol offers significant advancements for drug development.
  • This method allows for precise characterization of target engagement without molecular modification.
  • The protocol is particularly valuable for analyzing complex cellular systems and diverse cell types.