DigiWest® Protein Profiling Services

  • DigiWest® immuno-assays enable comprehensive analyses of up to 800 total & phosphoproteins using just ~20-60 µg of total protein, from 2D/3D cells, xenografts, tissues or even sections (e.g. LCM)

  • DigiWest® studies are fully customizable, based on a list of currently 1,208 antibodies, including 389 antibodies against phospho-epitopes and other post-translational modifications (PTMs), with 50+ pre-defined pathway panels for human, mouse, rat, dog, cyno & minipig samples

  • DigiWest® represents a versatile platform for pathway profiling, drug MoA studies, compound characterization (small molecules, PROTACs, antibodies, etc.), biomarker discovery, and safety/tox

  • DigiWest® combines the robustness of Western blotting with the power of Luminex® bead-based multiplexing and has been invented, developed and validated by Dr. Markus Templin's group at NMI

  • DigiWest® is being offered on a fee-for-service basis

  • New! Our new "Pathway Activity Panels" are designed to profile activation/ phosphorylation of key cell signaling pathways (RTK/RAS/RAF/ERK, PI3K/AKT, mTOR). As these pathways are frequently targeted by anti-cancer drugs, our Pathway Activity Panels are valuable for both clinical and pre-clinical samples. Contact us to get the list! And we have also just established DigiWest® for FFPE (formalin-fixed paraffin-embedded) samples ...
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 promising 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 as compared to reference drugs.

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 phenotypic differences in 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

Zhan T, et al. MEK inhibitors activate Wnt signaling and induce stem cell plasticity in colorectal cancer. Nature Communications 2019

Laner-Plamberger S, et al. H. differentially impacts gene expression of stromal cells from various tissues. Scientific Reports 2019

Christian S, et al. The novel dihydroorotate dehydrogenase (DHOH) inhibitor BAY 2402234 triggers differentiation and is effective in the treatment of myeloid malignancies. Leukemia 2019

Schumacher D, et al. Heterogeneous pathway activation and drug response modelled in colorectal-tumor-derived 3d cultures. PLoS Genetics 2019

Hoerst K, et al. Regenerative potential of adipocytes in hypertrophic scars is mediated by myofibroblast reprogramming. J Mol Med 2019

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

Inder S, et al. Profiling of radioresistant prostate cancer cells identifies key protein. BJUI 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