DigiWest High Content Protein Profiling

Imagine you get data from 80, 300, or even 600 total & phospho proteins per sample. With DigiWest you can!

DigiWest:
• Is a novel proprietary immunoassay technology
• Combines the robustness of Western blotting with the power of Luminex® bead-based multiplexing
• Allows for quantification of up to 600 analytes from <50 µg of protein
  
• Is fully customizable, based on a list of >1,000 antibodies (and validated antibody lists for human, mouse, rat, dog, minipig, cyno)
  
• Quantifies also post-translational modifications incl. phospho-epitopes
• Enables comprehensive analyses from limited samples (e.g. LCM)
• Is superior to MassSpec, needs less material & no enrichment
• Represents a versatile new platform for pathway profiling, drug MoA studies, lead characterization, predictive tox and biomarker screening
• Is being offered on a fee-for-service basis
  

Case Study 1: DigiWest for Biomarker Discovery

Analysis of mechanisms and biomarkers of Platinum resistance

• A set of 24 fresh frozen tumor specimens from relapsed vs cured platinum-treated ovarian cancer patients was analysed by DigiWest, using 466 antibodies (total and phospho proteins) covering various cell signalling pathways.
• DigiWest protein signatures distinguished platinum resistant vs sensitive patients, and revealed 8 biomarker candidates.
  

Case Study 2: DigiWest for Lead Characterization

Mode-of-action study of compounds in comparison to reference drugs

• Calu1 cells were treated with Trametinib (MEK inh) vs Copanlisib (PI3K inh) vs 2 experimental lead compounds (data not shown) vs DMSO. Cell samples were analysed by DigiWest, using 156 selected antibodies (total and phospho proteins) covering different signal transduction pathways. 
  
• DigiWest yielded distinct signatures for each compound, and allowed in-depth characterization of lead compounds.
  

Case Study 3: DigiWest in PD3D^TM Tumor Cell Models

Protein profiling for characterization of primary tumor samples 

• In collaboration with cpo cellular phenomics & oncology GmbH, primary patient-derived 3D tumor cell cultures (PD3Ds) were profiled by DigiWest, focusing on 122 antibodies for total and phospho proteins involved in a range of signaling pathways. 
  
• DigiWest revealed pathway activation patterns that explained differences in the PD3Ds' drug responses as measured by cpo.
  

Case Study 4: DigiWest in Multiple Species

Multiplex protein profiling for toxicology and in vivo pharmacology 

• A DigiWest panel of >700 antibodies previously validated in human and mouse materials was tested in protein lysates from liver and kidney samples of rat, dog, minipig and cynomolgus origin. 
• This comprehensive validation project has yielded hundreds of antibodies for DigiWest and Western blot studies, thus enabling protein profiling in toxicology and in vivo pharmacology.
  

References

Treindl F, Ruprecht B, Beiter Y, Schultz S, Döttinger A, Staebler A, Joos TO, Kling S, Poetz O, Fehm T, Neubauer H, Kuster B, Templin MF. A bead-based western for high-throughput cellular signal transduction analyses. Nature Communications 2016.

Augustin I, Dewi DL, Hundshammer J, Erdmann G, Kerr G, Boutros M. Autocrine Wnt regulates the survival and genomic stability of embryonic stem cells. Science Signaling 2017.