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bioRxiv · 10.64898/2025.12.12.693909

Evaluation of peptidomimetic inhibitors

Lenz, C.·Knapp, S.·Saxena, K.
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Abstract

Scientific research in drug discovery relies on robust, versatile, and well-established methodologies. One effective strategy for targeting disease-relevant proteins, such as E3 ligases, involves rational drug design using degron peptides as starting points for compound development. Here, we present a comprehensive set of complementary assays for the evaluation of degron-based binders, enabling their integration into an efficient drug discovery pipeline. We introduce screening strategies such as Differential Scanning Fluorimetry (DSF) and Fluorescence Polarization (FP), alongside both solution-based and immobilization-based biophysical techniques, including Isothermal Titration Calorimetry (ITC) and Surface Plasmon Resonance (SPR) for reliable affinity determination. To assess intracellular interactions with full-length target proteins, we also employ Nano Bioluminescence Resonance Energy Transfer (NanoBRET). Together, these methods establish a robust framework for the discovery and characterization of degron-based peptidomimetic compounds.

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Lenz, C., Knapp, S., Saxena, K.. 2025-12-12. Evaluation of peptidomimetic inhibitors. https://doi.org/10.64898/2025.12.12.693909

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