DigiWest High Content Protein Profiling for Target Identification

DigiWest® High-Content Protein Profiling Services

LI-COR and NMI TT Pharmaservices have partnered to bring you DigiWest® Protein Profiling, a high-content, multiplex immunoassay performed as a contract research service. The assay allows for the quick and efficient discovery of biomarkers or characterization of potential targeted therapeutics. A DigiWest® assay provides comprehensive biological information deep into one or several cell signaling pathways.

The information gained from a DigiWest® Protein Profiling assay can help determine which targets or pathways offer the most therapeutic promise. The service can also be used to make predictions about the safety and toxicity of a therapeutic based upon functional effects and performance.

Watch to see how DigiWest® is making an impact in biomedical decision making and patient care.

High Content Screening with Efficient Sample Use

Only 20 to 60 μg of total protein are required to perform a DigiWest® assay. Using just 30 μg of snap-frozen tissue or dry-frozen cells, comprehensive analyses can be done using up to 800 different antibodies. A library of over 1,200 prevalidated antibodies and over 50 predefined pathways are available for human, mouse, rat, dog, minipig, and cynomolgus samples.

Uses for DigiWest® Protein Profiling Services

A DigiWest® immunoassay can provide data on several parameters useful in the development of targeted therapeutics.

  • Biomarker discovery to identify which biomarkers are promising targets for a therapeutic.
  • Lead characterization or mode-of-action study to provide insight into how a therapeutic works.
  • Predictive safety and toxicology investigation to estimate how safe a therapeutic might be based upon its functional effects.

Biomarker Discovery with DigiWest®

This study used DigiWest® for analysis of mechanisms and biomarkers of platinum resistance in ovarian cancer patients.

A set of 24 fresh frozen tumor specimens from relapsed vs cured platinum-treated ovarian cancer patients was analyzed by DigiWest®, using 466 antibodies (total and phospho proteins) covering various cell signaling pathways.

DigiWest® protein signatures distinguished platinum resistant vs sensitive patients, and revealed 8 promising biomarker candidates.

figure 1
Figure 1. Image shows a heat map of a subset of the profiled protein analytes (targeted with antibodies shown on the y-axis) in each tumor sample (x-axis). Red squares indicate an increase in protein expression compared to control, black indicates no change, and green squares indicate a decrease.

DigiWest® for Lead Characterization

In this study, DigiWest® was used to perform a mode-of-action study of compounds in comparison to reference therapeutics.

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 analyzed by DigiWest®, using 156 selected antibodies (total and phospho proteins) covering different signal transduction pathways.

DigiWest® yielded distinct signatures for each compound and allowed for in-depth characterization of lead compounds as compared to reference therapeutic.

figure 2
Figure 2. Distinct signatures for MEK vs PI3K inhibitors in Calu1 cells are shown using the heat map. Each horizontal line indicates a different antibody. Red lines indicate an increase in protein expression compared to control, black indicates no change, and green lines indicate a decrease.

Advantages of DigiWest® Protein Profiling Services

The analyses from a DigiWest® Protein Profiling assay provide a more complete understanding of how a therapeutic may affect a target and creates a potential new starting point in the therapeutic development process.1

  • High content
  • Low sample use
  • Library of over 1,200 prevalidated antibodies
  • 50+ predefined pathways
  • Targets total and phosphorylated proteins

Contact us to arrange a consultation with the DigiWest® Service Lab.

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  1. Kissel, M., Berndt, S., Fiebig, L., Kling, S., Ji, Q., Gu, Q., Lang, T., Hafner, F.T., Teufel, M., and Zopf, D. (2017). Antitumor effects of regorafenib and sorafenib in preclinical models of hepatocellular carcinoma. Oncotarget, 8(63), 107096–107108. https://doi.org/10.18632/oncotarget.22334