DigiWest® Protein Profiling Services

  • DigiWest® is a proprietary immunoassay technology
  • Allows analyses of up to 800 total and phospho proteins from just ~60 µg of total protein
  • Combines the robustness of Western blotting with the power of Luminex® bead-based multiplexing
  • Is fully customizable, based on a list of >1,000 antibodies, including >260 phospho-epitopes and other post-translational modifications (PTMs)
  • Comes with 50+ pre-defined pathway panels for human, mouse, rat, dog, cyno & minipig samples
  • Enables comprehensive analyses from limited samples (e.g. LCM)
  • Has been invented, developed and validated by Dr. Markus Templin's group at NMI
  • Represents a versatile platform for pathway profiling, drug MoA studies, lead characterization, predictive safety/tox and biomarker discovery
  • Is being offered on a fee-for-service basis
Logo of DigiWest - High Content Protein Profiling

Imagine to get data on 80, 300, or even 800 total & phospho proteins from just 10-60 micrograms of protein sample. With DigiWest® you can!

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.

 

Image of DigiWest - Biomarker Discovery

Case study 2: DigiWest® for lead characterization

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

  • Calu1 cells were treated with 1 MEK inhibitor vs 1 PI3K inhibitor 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.

 

Image of DigiWest - Lead Characterization

Case study 3: DigiWest® in PD3D® tumor models

Protein profiling for characterization of primary tumor samples

  • In collaboration with cpo Cellular Phenomics & Oncology GmbH, primary patient-derived 3D tumor cell cultures (PD3D®) 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 PD3D® drug responses as measured by cpo.

 

Image of DigiWest in PD3D Tumor Cell Models

Case study 4: DigiWest® in multiple species

Multiplex protein profiling for safety/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 safety/toxicology and in vivo pharmacology.

 

Image of DigiWest Protein Profiling in Multiple Species

Peer-reviewed publications

Sikorski K, et al. A high-throughput pipeline for validation of antibodies. Nature Methods 2018

Mehul B, et al. Mass spectrometry and DigiWest technology emphasize protein acetylation profile from [...] treated HuT78 CTCL cell line. Journal of Proteomics 2018

Lomnytska M, et al. Platelet protein biomarker panel for ovarian cancer diagnosis. Biomarker Research 2018

De Boussac H, et al. Epidermal Growth Factor represses CAR expression in primary hepatocytes and favors regulation by PXR. Drug Metab Dispos 2017

Kissel M, et al. Antitumor effects of [...] in preclinical models of hepatocellular carcinoma. Oncotarget 2017

Mishra M, et al. Protein purification and analysis: next generation Western blotting techniques. Expert Rev Proteomics 2017

Menck K, et al. Neutral sphingomyelinases control extracellular vesicles budding from the plasma membrane. J Extracell Vesicles 2017

Augustin I, et al. Autocrine Wnt regulates the survival and genomic stability of embryonic stem cells. Science Signaling 2017

Treindl F, et al. A bead-based western for high-throughput cellular signal transduction analyses. Nature Communications 2016

Other references

DigiWest: Multiplex profiling of proteins for biomarker discovery. LabsExplorer 2018

Sachse C, Templin M. Multiplexing Western blots: More data, less sample. GEN 2018

Erdmann G, et al. Towards 3Rs: Multiplex protein profiling from minipig, cyno, dog and rat samples, to get the most our of preclinical safety studies. Ellegaard Newsletter 2018

Graham H. Drop the blot: Parallel quantification of up to 800 proteins. Luminex Blog 2018

Singh G. The future of now - how the science of today shapes tomorrow. ScienceExchange Blog 2018

Pearson S. Screening innovations and 3D assay developments showcased at ELRIG. GEN 2017

Sachse C, Regenbrecht CRA, Templin MF. Beyond Genomics. DigiWest-Technologie: Multiplex-Analyse von Proteinen und Phosphoproteinen. LABO 2017

How to find the missing link between genotype and phenotype. Labiotech 2017

Evans J. A blot on the beads: Western blotting with a bead-based immunoassay. Electrophoresis 2016

Couvillon A, Templin MF. Cross-validation of antibodies using DigiWest. LabRoots 2016

Graham H. DigiWest: Reimaging Western blotting on the proteomic scale. Luminex 2016