Ultrafast and Predictive Mass Spectrometry-Based Autotaxin Assays for Label-Free Potency Screening

SLAS Discov. 2017 Apr;22(4):425-432. doi: 10.1177/2472555217690484. Epub 2017 Feb 2.


Autotaxin (ATX) is a promising drug target for the treatment of several diseases, such as cancer and fibrosis. ATX hydrolyzes lysophosphatidyl choline (LPC) into bioactive lysophosphatidic acid (LPA). The potency of ATX inhibitors can be readily determined by using fluorescence-based LPC derivatives. While such assays are ultra-high throughput, they are prone to false positives compared to assays based on natural LPC. Here we report the development of ultrafast mass spectrometry-based ATX assays enabling the measurement of data points within 13 s, which is 10 times faster than classic liquid chromatography-mass spectrometry. To this end, we set up a novel in vitro and whole-blood assay. We demonstrate that the potencies determined with these assays are in good agreement with the in vivo efficacy and that the whole-blood assay has the best predictive power. This high-throughput label-free approach paired with the translatable data quality is highly attractive for appropriate guidance of medicinal chemists for constructing strong structure-activity relationships.

Keywords: LPA assay; RapidFire; autotaxin; label free; mass spectrometry.

MeSH terms

  • Animals
  • Dogs
  • Enzyme Inhibitors / blood*
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology
  • Haplorhini
  • High-Throughput Screening Assays*
  • Humans
  • Hydrolysis
  • Lysophosphatidylcholines / blood*
  • Lysophosphatidylcholines / chemistry
  • Lysophospholipids / antagonists & inhibitors
  • Lysophospholipids / blood*
  • Lysophospholipids / chemistry
  • Mass Spectrometry / methods*
  • Phosphoric Diester Hydrolases / blood*
  • Rats
  • Rats, Wistar
  • Recombinant Proteins / blood


  • Enzyme Inhibitors
  • Lysophosphatidylcholines
  • Lysophospholipids
  • Recombinant Proteins
  • Phosphoric Diester Hydrolases
  • alkylglycerophosphoethanolamine phosphodiesterase