In the forefront of modern therapeutic innovation, Targeted Protein Degradation (TPD) represents a groundbreaking approach that is reshaping the landscape of drug discovery. Unlike traditional methods that inhibit protein function, TPD leverages the body's natural cell machinery to selectively degrade and eliminate disease-causing proteins. This revolutionary technique offers a new horizon for tackling previously undruggable targets, opening vast opportunities for the development of novel treatments for a range of intractable diseases.
At ICE Bioscience, our TPD Services are designed to empower your drug discovery journey, offering comprehensive solutions that span from target identification to the development of bespoke degraders. Leveraging cutting-edge technology, deep scientific expertise, and a collaborative approach, we are committed to advancing your projects with precision and efficiency. Whether it’s cancer, neurodegenerative disorders, or other complex conditions, our TPD platform is equipped to meet the challenges of modern medicine, driving forward the discovery of transformative therapeutic agents.
| Category | Solutions | Assays |
|---|---|---|
| In vitro Biology | Protein production | Target protein, E3 |
| Target degradation validation | E3 OE/KD/KO (siRNA, shRNA, sgRNA)TPD leverages cellular protein degradation pathways to selectively degrade disease-related target proteins. Within cells, two primary degradation mechanisms exist: the ubiquitin-proteasome system (UPS) and the lysosomal pathway. A key component of the UPS is the E3 ubiquitin ligase, which labels target proteins with ubiquitin, directing them to the proteasome for degradation. One notable E3 ligase is VHL (von Hippel-Lindau), commonly used in TPD studies for its role in targeting hypoxia-inducible factors. ICE Bioscience has established VHL knockout (KO) cell lines specifically designed to support VHL-related screening and validation assays, enabling efficient evaluation of TPD strategies that involve VHL and related pathways. | |
| Target and POI Degradation VerificationTarget validation is a critical step to confirm that the drug interacts with the intended molecular targets and that these targets are directly relevant to the disease. ICE Bioscience offers a range of methods for thorough target validation, including gene knockout and silencing, multi-omics approaches, and phenotypic screening. We employ CRISPR-Cas9 and other gene-editing technologies to knock out or silence specific genes, enabling the observation of cellular function and phenotype changes to support robust target validation. Additionally, we provide target protein degradation verification using HiBiT and NanoLuc technologies. Our team constructs plasmids for expressing HiBiT/NanoLuc-fusion proteins, applicable through transient or viral transduction. This advanced plasmid system allows for rapid and precise screening of degradation agents, streamlining the discovery and validation of effective degradation compounds. | ||
| Protein t1/2 Protein half-life reflects the time required for a protein’s concentration to degrade to half its initial level, indicating its stability and dynamic balance within the cell. ICE Bioscience offers multiple assays for accurate half-life measurement: 1. CHX Inhibition Assay: We add cycloheximide (CHX) to block new protein synthesis and track protein degradation over time using standard detection techniques like Classic WB, Simple Western (Jess), In-Cell Western, and IF. 2. Doxycycline (DOX)-Inducible System: Our team has developed an inducible plasmid system for rapid half-life determination. By inducing protein expression to the plateau phase with DOX, we then halt induction and monitor natural degradation using HiBiT, calculating protein half-life through curve fitting. | ||
| Warhead optimization & screening | E3 binder screening: TR-FRET, SPR | |
| POI binder screening: TR-FRET, SPR, TSA, FI, FP, Luminescence, AlphaLISA, LCMS, etc. | ||
| Ternary complex formation assay: TR-FRET, AlphaLISA, SPR, Spectral Shift-TRIC, NanoBRET, etc. | ||
| Cellular degradation and functional assay | WB/Simple Western (Jess) | |
| HiBiT-KI cell line construction & HiBiT screening | ||
| In-Cell Western/Immunofluorescence Assay/HTRF/AlphaLISA/MSD/Flow Cytometry/ELISA | ||
| Quantitative Proteomics & Proximity Proteomics | ||
| Cell viability (2D/3D): Cell proliferation, Clonogenic assay | ||
| Downstream signaling protein detection | ||
| Immune functional assays | ||
| Reporter gene assays | ||
| Cell cycle and cell apoptosis assays | ||
| Cell panel screening | ||
| Mechanism & permeability study | Degradation kinetics Our degradation kinetics testing service provides a comprehensive approach to evaluating protein degradation over time in response to TPD molecules. Using a variety of advanced assays — including Western Blot (WB), Simple Western (Jess), HiBiT, Flow Cytometry, HTRF, and AlphaLISA — we measure protein levels at multiple time points post-treatment to capture precise degradation dynamics. | |
| Degradation pathway: Proteosome or lysosomeOur Degradation Pathways Testing service provides insights into the cellular pathways responsible for drug-induced protein degradation, including the ubiquitin-proteasome system (UPS) and lysosomal pathways. By pre-treating cells with proteasome or lysosome inhibitors before applying TPD molecules, we can pinpoint which degradation pathway is involved. Comparative degradation analysis, with or without inhibitors, allows us to determine the pathway through which proteins are targeted. Our assays utilize advanced detection techniques, such as WB, Simple Western (Jess), ELISA, AlphaLISA, HTRF, Flow Cytometry, and MSD, ensuring precise and comprehensive pathway profiling. | ||
| Proximity-based cellular assay: NanoBRET, NanoBiT, TR-FRET, Co-IP | ||
| Ubiquitination assayOur Ubiquitination Assay service provides detailed analysis of how drugs induce ubiquitination, a crucial step in TPD. Ubiquitination typically involves an E1-E2-E3 enzyme cascade, with E3 ligases specifically recognizing and tagging target proteins for degradation. To confirm whether a protein is targeted for degradation, we assess its ubiquitination status using assays, such as Western Blot, ELISA. We also offer TR-FRET assays to detect ubiquitin binding to the protein of interest (POI) and NanoBRET technology for real-time ubiquitination analysis in live cells. | ||
| Off-Target Studies for Selectivity AssessmentOur Off-Target Studies service is designed to evaluate non-specific degradation effects, commonly known as off-target effects, in drug development. By comparing the overall proteome and ubiquitinated proteome, we can identify proteins affected unintentionally by the drug, ensuring its specificity and reducing potential side effects. We employ advanced techniques, including Simple Western (Jess), In-Cell Western, and proteomics, to assess selectivity and off-target impacts accurately. | ||
| NanoBRET target engagement assay (Live & permeabilized) |
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