A high content, high throughput cellular thermal stability assay for measuring drug-target engagement in living cells
作者: Andrew J. Massey
作者单位: 1Vernalis Research, Granta Park, Cambridge, United Kingdom
刊名: PLOS ONE, 2018, Vol.13 (4)
来源数据库: PLOS
DOI: 10.1371/journal.pone.0195050
原始语种摘要: Determining and understanding drug target engagement is critical for drug discovery. This can be challenging within living cells as selective readouts are often unavailable. Here we describe a novel method for measuring target engagement in living cells based on the principle of altered protein thermal stabilization / destabilization in response to ligand binding. This assay (HCIF-CETSA) utilizes high content, high throughput single cell immunofluorescent detection to determine target protein levels following heating of adherent cells in a 96 well plate format. We have used target engagement of Chk1 by potent small molecule inhibitors to validate the assay. Target engagement measured by this method was subsequently compared to target engagement measured by two alternative methods...
全文获取路径: PLOS  (合作)
影响因子:3.73 (2012)

  • engagement 啮合
  • target 目标
  • cells 麻风细胞
  • throughput 吞吐量
  • measuring 测量
  • thermal 热的
  • protein 蛋白质
  • adherent 粘附的
  • determination 测定
  • cellular 细胞状的