Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay

Science. 2013 Jul 5;341(6141):84-7. doi: 10.1126/science.1233606.


The efficacy of therapeutics is dependent on a drug binding to its cognate target. Optimization of target engagement by drugs in cells is often challenging, because drug binding cannot be monitored inside cells. We have developed a method for evaluating drug binding to target proteins in cells and tissue samples. This cellular thermal shift assay (CETSA) is based on the biophysical principle of ligand-induced thermal stabilization of target proteins. Using this assay, we validated drug binding for a set of important clinical targets and monitored processes of drug transport and activation, off-target effects and drug resistance in cancer cell lines, as well as drug distribution in tissues. CETSA is likely to become a valuable tool for the validation and optimization of drug target engagement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antimetabolites, Antineoplastic / metabolism
  • Cell Line, Tumor
  • Dose-Response Relationship, Drug
  • Drug Monitoring / methods*
  • Folic Acid Antagonists / metabolism
  • Hot Temperature*
  • Humans
  • Kidney / metabolism
  • Ligands
  • Liver / metabolism
  • Molecular Targeted Therapy*
  • Pharmaceutical Preparations / metabolism*
  • Protein Binding
  • Protein Stability
  • Proteins / metabolism*
  • Quinazolines / metabolism
  • Thiophenes / metabolism
  • Tissue Distribution


  • Antimetabolites, Antineoplastic
  • Folic Acid Antagonists
  • Ligands
  • Pharmaceutical Preparations
  • Proteins
  • Quinazolines
  • Thiophenes
  • raltitrexed