Ready-To-Go Assays

Cutting-Edge & Cost-Efficient Services For Multiple Drug Targets

Biophysical & Biochemical Services to Characterize Target Interaction

Ready to go, off the shelf assays, biophysical and biochemical, target identification and validation

Established Assays

Optimized assay conditions

Ready to go, off the shelf assays, biophysical and biochemical, target identification and validation, parallel

Flexible

Single/multiple assays in parallel

Ready to go, off the shelf assays, biophysical and biochemical, target identification, cost-effective

Cost-effective

High quality data

Ready to go, off the shelf assays, biophysical and biochemical, rapid, quick turnaround times, TAT

Fast Turnaround Time

Less than 2 weeks

nanoDSF – MST – SPR – ADP-Glo – Nucleotide exchange

Protein Supply

Established protein constructs in-house

Tailored high quality assay grade  proteins

Protein QC: SDS-PAGE, aSEC, MS

Complete protein supply included

Assay Initiation & Measurements

Established assay conditions

Established labeling/coupling method

Published tool compounds

Up to 3 Tm/ Kds / IC50s included

Results & Report

A tabulated report included

Extended report available including  measured curves, assay conditions,  analysis and scientific summary in  pdf-format

Biophysical Assays covering a Broad Range of Drug Targets

TargetUniProt IDPPMSTnanoDSFSPRBiochemical Assay
CDK7P50613_
CRBN/DDB1Q96SW2/Q16531developing_
cRELQ96HD1__
DOCK5Q9H7D0nucleotide exchange
GLUT1P11166__
GRB2P62993__
KRASP01116nucleotide exchange
MALT1Q9UDY8_
MAP4K1Q92918developingADP Glo
NLRP3Q96P20developingADP Glo
NRASP01111nucleotide exchange
P2RX3P56373__
p38Q15759__
PolQO75417__
SHP2Q06124__
SMARCA2P51531developingADP Glo
SOS1Q07889_nucleotide exchange
STAT3P40763__
STAT4Q14765___
STAT6P42226_
STINGQ86WV6_
USP7Q93009_
WRNQ14191_

Case Study: biophysical assays to identify novel K-RAS inhibitors

K-RAS is a member of the RAS protein family – GTPases which are involved in signaling processes leading to cell growth and proliferation. Oncogenic variants contribute to 25% of cancers including colorectal and lung cancer. This makes the RAS family of proteins important targets for cancer treatment. After 30 years of research, the recently developed K-RAS inhibitor (Canon et al., 2019) opened the door for promising strategies to target this protein family.

To support and accelerate drug development campaigns to identify novel K-RAS inhibitors, WuXi AppTec offers a platform for small molecule hit-finding and lead optimization including biophysical assays and structure biology services based on established systems.

Protein Production

  • Broad variety of constructs for biophysical assays and crystallography established including,
    • wild type
    • G12C, G12D, G12V
    • G13D
    • “Cys-light” variants for covalent inhibitors
  • Nucleotide exchange protocols established to provide K-RAS variants loaded with GDP and alternative nucleotide analogues such as GTPyS or GMPPNG GDP
  • Characterization and optimization of covalent complex formation using HPLC-MS
  • Related targets protein interaction partners such as SOS1 and Raf available
KRAS, K-RAS inhibitors, assay-grade and crystal grade proteins, G12C, G12D, GTPase

Biophysical assays for different K-RAS variants established as “Ready-to-Go”

nanoDSF

nano-DSF, nanoDSF biophysical assays for KRAS variants

Nucleotide exchange assay

Nucleotide exchange assays, KRAS, K-RAS, SOS, ready-to-go

Labeled MST

Labeled MST assessment

GFP-KRAS Fusion

GFP-KRAS fusion protein, MST

MST in eukaryotic cell lysates

MST in eukaryotic cell lysates

Crystallography

  • Established co-crystallization system for covalent and non-covalent small molecule inhibitors
  • Non-covalent and covalent inhibitors; GDP and non-hydrolysable GTP analog bound states
Crystallography, co-crystallization for covalent and non-covalent small molecule inhibitors, crystal grade protein
  • Rapid data collection – K-RAS data sets up to 1.3 Å resolution collected in-house on a Bruker MetalJet
  • Established crystallization system for related targets e.g.: SOS1, K-RAS-SOS1 complex
Crystal-grade protein, SOS1, K-RAS-SOS1 complex, high resolution, co-crystallization