Cell-based Epigenetic Assay Services

Creative Biolabs provides cell-based epigenetic assay services testing the modulation of histone deacetylation, histone methylation and histone phosphorylation. Numerous cell-based methods that our scientists have created should make it easier to chemically screen for and profile these epigenetic targets. This will help guide research programs in the development of novel chemotypes with higher cellular potency and specificity.

In vitro biochemical assays are typically the first option for routine primary screening because they can be easily automated and miniaturized. Even though a lot of biochemical experiments have been modified to use nucleosomes, octamers, and whole histone proteins as substrates, these modifications are unable to accurately replicate native circumstances. Since many epigenetic enzymes exist in cells as complexes of many regulatory subunits, it may be challenging to express them as functional enzymes that can be reconstituted. Furthermore, the impact of other nearby histone posttranslational modifications on the methylation of histone tails remains poorly known. These restrictions are overcome by cell-based assays, which measure histone modifying enzymes in their native habitat and with appropriate substrates.

Fig.1 The enzyme (yellow) interacts (red arrows) with several proteins inside the cell.¹

Choosing a Cell-based Epigenetic Assay for Compounds Profiling

• Antibody-based assays
  Details: Nuclear extraction followed by Western blot or a sandwich immunoassay using histone methylation-specific antibodies is a conventional antibody-based assay. A reasonably sensitive way to identify changes in histone methylation levels globally is made possible by the availability of commercial histone methylation ELISA kits.
  Format: 384-well microplates
  Targets: HDAC, Methyltransferase, Kinases
• Binding assays
  Details: In addition to reporting on inhibitor cell permeability, target engagement, and intracellular potency, cellular ligand binding platforms are effective methods for determining a compound's ability to bind to its protein target in its native cellular context. These plates depend on the well-characterized characteristic of protein targets to modify their biophysical characteristics upon attachment to tiny ligands.
  Format: 384-well microplates
  Targets: Kinases
• Mass spectrometry-based assays
  Details: Enzymatic activity in response to small molecule modulation is measured in vitro using mass spectrometry (MS) based methods. There has been a study on an MS-based technique that analyzes JMJD2A inhibition in cells by tracking H3K9me3 levels. The substrate that JMJD2A prefers. The ultraperformance liquid chromatography high/low collision switching–MS (UPLC–MS) was used to examine the results.
  Targets: Histone methyltransferases (HMTs) and demethylases (HDMs)

Few compounds are presently in advanced phases of clinical studies, despite the mounting body of evidence indicating the causative contribution of the expanding number of epi-based inhibitors exhibiting activity against human illnesses. Drug combinations and time-dependent dosages are crucial to the clinical development of these medications. The variety of possible therapeutic alternatives expands rapidly with the ability to arrange the continuous administration of molecules and use different dosages. Creative Biolabs expedites the drug development process with cell-based epigenetic assay services. Please contact us for more information.

Reference

1. Benedetti, Rosaria, et al. "Epigenetic-based therapy: From single-to multi-target approaches." The international journal of biochemistry & cell biology 69 (2015): 121-131.