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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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References

  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

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