Poly(ADP-ribose) Polymerase (PARP) Panel Screening Service

We specialize in high-throughput screening (HTS) to efficiently identify novel PARP inhibitors from vast compound libraries. Following initial hit identification, our services extend to comprehensive compound profiling, including the determination of half-maximal inhibitory concentration (IC50) values and detailed selectivity assessments across various PARP isoforms. This meticulous approach is crucial for minimizing off-target effects and identifying compounds with optimal therapeutic windows. We also offer lead generation and structure-activity relationship (SAR) studies to systematically optimize compound potency and pharmacokinetic properties.

Our service portfolio incorporates a diverse range of cutting-edge assay technologies, ensuring robust and reproducible results:

• Biochemical Activity Assays: These in vitro assays quantify PARP enzymatic activity by measuring the consumption of NAD+ or the transfer of ADP-ribose to histone proteins. Our capabilities include:
  • Fluorescence Polarization Assays: Utilized for PARP trapping, where an inhibitor prevents ribosylation and effectively traps PARP onto a fluorescent oligonucleotide duplex, leading to a measurable increase in signal.
  • Bioluminescent Assays: These homogeneous, add-and-read formats are ideal for HTS, detecting NAD(+) and NADH levels consumed during PARP activity or measuring AMP produced from enzymatic reactions.
  • Kinase Assays: While primarily for kinases, this universal biochemical assay can be adapted to monitor the activity of enzymes that produce ADP, including certain PARPs, and is valuable for inhibitor profiling and mode of action studies. Our standard biochemical assays typically utilize histone substrate and biotin-labeled NAD+, with activity detected by adding Strep-horseradish peroxidase (HRP) and a chemiluminescent substrate, ensuring high sensitivity and reliability.
• Cell-Based Target Engagement Assays: For a more physiologically relevant assessment, we employ live-cell assays:
  • Target Engagement Assays: This powerful biophysical method assesses compound binding to target proteins directly within live cells using bioluminescent resonance energy transfer.
  • Immunoassay Cellular Systems: These no-wash bioluminescent immunoassays are designed to measure target analytes directly in cell lysates or culture media, making them ideal for secondary screening and lead optimization.
  • Cell Viability Assays: We utilize sensitive assays for measuring cell health and cytotoxicity in live cells, including 3D cultures, which are scalable for high-throughput primary screening.

Creative Biolabs boasts access to one of the largest PARP protein panels on the market, enabling comprehensive efficacy comparisons across a broad spectrum of the PARP family. This extensive panel, encompassing over 10 different PARP isoforms, allows for unparalleled selectivity profiling. For every project, clients receive detailed reports including raw data, percentage enzyme activity, and control compound IC50 values in an easily accessible Excel format.