As one of the important classes of drug target, Kinase remains an area of interest for both pharmaceutical industry and academia. To assist the efforts in kinase drug discovery, WuXi Biology offers high quality, time efficient and cost-effective kinase solution for assay development, high throughput screening (HTS) and selectivity profiling. WuXi Biology provides multiple cellular assay development formats to support different functional assessments of the drug candidates.

Assay development

Assay development is applicable for a broad range of kinases involved in MAPK pathway, cell cycle, NF-kB pathway, AKT/PKB pathway, JAK/STAT, membrane receptor, etc.

  • Biochemical assay:
    • A variety of formats: TR-FRET (LANCE Ultra, LathaScreen, HTRF), Caliper, Luminescence (Kinase-Glo, ADP-Glo), LC-MS, Radiometric, IMAP, ADP-Quest, Z’ LYTE, etc.
    • Development includes routine assay optimization (enzyme titration, time course, Km determination etc.) and assay validation (DMSO tolerance, Z’ test, compound validation, etc.).

WuXi Biology also provides service for compound MOA study in the effort to further characterize the identified potent compounds.

Results from various biochemical assay development for protein kinases.
(A) Establishment of initial velocity for the determination of kinase A ATP Km.
(B) ATP Km determination for kinase A using ADP Quest format.
(C) Kinase profiling-IC50 determination using Kinase Glo Plus.
Curve fitting was performed using GraphPad Prism.

ELISA Potency Assay

  • Cellular Assay:
    • Activity evaluation: ELISA, AlphaLISA, Western Blot, LanthaScreen activity assay, Cisbio HTRF assay, Pathhunter kinase assay
    • Compound binding assay: ActivX assay, LanthScreen binding assay
    • Protein interaction detection: Cisbio HTRF, PathHunter binding assay
    • Cellular selectivity screen: ActivX assay, MSD
  • Compound Screening:
    • We support HTS and routine SAR screening against enzyme targets using either WuXi Biology developed assays or assays transferred from clients.

Kinase Selectivity Profiling Services

  • Profiling format: Caliper, Kinase Glo Plus, ADP Glo
    • The diversified panel for profiling services.
    • Single dose measurement and IC50 determination
    • Single compound test and library screening
    • Data turnaround time: 1-2 weeks
Compound A selectivity profile against a panel of 30 kinases.