DigiWest High Content Protein Profiling | NMI TT Pharmaservices

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NMI-TT Contact Person

Dr. Markus Templin

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DigiWest: High Content Protein Profiling Services

DigiWest enables multiplexed protein profiling of up to 600 analytes per sample, thus increasing Western Blot outputs by up to 300-fold.

    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 PD3DTM 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.

 

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