Protac® In Vitro Evaluation
We provide comprehensive PROTAC in vitro evaluation services, covering a variety of detection methods from protein level to cell level, including binary and ternary complex formation analysis, target ubiquitination analysis, target degradation level analysis, cytotoxicity experiments, cell function impact assessment and intracellular protein degradation detection, etc. With a professional technical team, a complete analysis platform and a strict quality control system, we ensure the accuracy and reliability of the data and provide strong support for customers' PROTAC molecule research and development.
Our Available PROTAC In Vitro Evaluation Services
Protein Level Detection Services
Binary and Ternary Complex Formation Analysis: Detect the binary and ternary complexes formed between PROTAC molecules, target proteins, and E3 ligases, and evaluate their binding abilities.
Target Ubiquitination Analysis: Analyze the level of target protein ubiquitination induced by PROTAC molecules to verify their ability to effectively mediate the ubiquitination process of target proteins.
Protein Degradation Kinetic Study: Study the kinetic characteristics of PROTAC molecules on target protein degradation using methods such as Western blot, In-cell western, and HTRF, including degradation efficiency, specificity, dose-dependency, and time-dependency (e.g., DC50 and Dmax parameters).
Cellular Level Detection Services
Cytotoxicity Assay: Use methods like CCK-8 or CTG to assess the inhibitory effect of PROTAC molecules on cell proliferation and analyze their toxicity on cancer cells.
Cell Function Impact Evaluation: Evaluate the effects of target protein degradation on cell functions, such as changes in cell proliferation, apoptosis, and migration ability.
Intracellular Protein Degradation Detection: Use technologies like HiBiT-tagged POI stable cell lines to conduct high-throughput screening and detect the intracellular degradation ability of PROTAC molecules on target proteins.
Screening and Optimization Services
PROTAC Molecule Screening: Use affinity screening technologies (such as ASMS, SPR, etc.) to screen for high-affinity PROTAC molecules from compound libraries.
High-Throughput Screening Platform: Use "endogenous protein-HiBiT" stable cell lines created by gene knock-in technology to conduct high-throughput screening of PROTAC molecules and calculate DC50 values.
Lead Molecule Optimization: Support library construction, high-throughput screening, and lead molecule optimization needs to improve the structure and performance of PROTAC molecules.
Pharmacokinetic Research Services
In Vitro Transporter Study: Study the transporter properties of PROTAC molecules in vitro, such as permeability and transporter inhibition effects.
Metabolic Stability Study: Evaluate the metabolic stability of PROTAC molecules in vitro, including liver cell metabolism stability tests, plasma protein binding tests, and more.
Metabolite Identification: Identify and analyze the metabolites of PROTAC molecules through various metabolite identification models.
Biological Sample Analysis: Perform biological sample analysis of PROTAC molecules using LC-MS/MS platforms, offering bioanalysis services throughout the preclinical stages of drug development.
Other Services
E3 Ligase Analysis: Analyze the expression and function of potential E3 ligases to provide guidance for PROTAC molecule design.
Target Protein Degradation Validation: Validate the degradation effect of PROTAC molecules on target proteins using various methods to ensure mechanism reliability.
Omics Analysis: Use transcriptomics and proteomics to reveal the global gene and protein expression effects of PROTAC molecules.
Why Choose Our Services?
Technical Expertise: A professional technical team skilled in advanced PROTAC in vitro evaluation techniques, providing comprehensive and precise testing services.
Platform Integrity: A complete PROTAC in vitro analysis platform covering a wide range of detection methods from protein level to cell level, offering comprehensive solutions to clients.
Method Diversity: Provide various detection methods such as Western blot, In-cell western, and HTRF, allowing clients to choose the most suitable methods according to their needs and project characteristics.
Data Reliability: A strict quality control system ensures the accuracy and reliability of experimental data, providing strong support for clients' R&D decisions.
Customized Services: Provide personalized service plans based on clients' specific needs, meeting the unique requirements of different projects.
Project Experience: Rich project experience in PROTAC in vitro evaluation, enabling quick and efficient completion of projects of various scales.
Innovation: Stay up-to-date with the latest developments in PROTAC technology, continuously optimizing and innovating service offerings to provide clients with cutting-edge technical support.
One-Stop Service: We also offer one-stop services from molecular design to in vivo efficacy validation, helping clients develop next-generation precision-targeted drugs.
Workflow of PROTAC In Vitro Evaluation
- Project Consultation and Communication: Initial communication between the client and service provider to clarify project requirements, goals, and expected outcomes.
- Scheme Design: The service provider creates a detailed experimental plan based on the client's needs, including experimental methods, sample requirements, and scheduling.
- Sample Preparation: The client prepares samples as required, such as cell lines and protein samples, and submits them to the service provider.
- Experimental Implementation: The service provider conducts the PROTAC in vitro evaluation experiments as per the experimental plan, including protein level detection, cellular level detection, etc.
- Data Analysis and Interpretation: Analyze and interpret the experimental data, evaluating PROTAC molecule activity, specificity, degradation efficiency, and other indicators.
- Report Writing and Submission: Prepare a detailed experimental report, including experimental methods, results, data analysis, and conclusions, and submit it to the client.
- Follow-up Support: Provide subsequent technical support and consultation services based on client feedback and needs.
Technology Platform
Protein Level Detection Platform: Using affinity detection technology and ternary complex formation analysis, this platform accurately evaluates the binding ability of PROTAC molecules with target proteins and E3 ligases, as well as the formation of complexes.
Cellular Level Detection Platform: Utilizing protein degradation detection technology and cell function impact assessment techniques, this platform efficiently detects the degradation efficiency of PROTAC molecules on target proteins and their effects on cellular functions.
Screening and Optimization Platform: Combining high-throughput screening technology and computational drug design platforms, this platform rapidly identifies and optimizes PROTAC molecules with high degradation activity, accelerating the drug development process.
Pharmacokinetic Research Platform: Through in vitro transporter study models, metabolite identification models, and biological sample analysis platforms, this platform deeply investigates the metabolic properties, transport characteristics, and content changes of PROTAC molecules in biological samples.
Frequently Asked Questions (FAQs)
1. What are the experimental methods for PROTAC molecule in vitro evaluation services?
- Binary and ternary complex formation analysis.
- Target ubiquitination analysis.
- Target degradation level analysis.
- Cytotoxicity assays.
- Cell function impact evaluation.
- Intracellular protein degradation detection.
2. Does poor in vitro permeability of a PROTAC molecule indicate poor in vivo absorption?
PROTAC molecules that show low permeability in vitro may still achieve moderate bioavailability in animals. Therefore, it is advised to use in vitro permeability data cautiously when screening PROTAC compounds.
3. What early DMPK studies are recommended for PROTAC molecules?
One part is used to characterize the basic DMPK properties of the compound, including determining physicochemical properties, conducting in vitro permeability testing, plasma protein binding, and preliminary in vitro drug interaction tests.
Another part focuses on further screening and optimization of the compound, with emphasis on solubility determination and in vitro metabolic studies (including both qualitative and quantitative studies).
4. Do metabolites with cleavage-type structures in PROTAC molecule metabolism studies require additional toxicology studies?
If certain conditions are met, it is recommended to conduct toxicological evaluation of the metabolites.
5. How to evaluate the protein degradation selectivity of PROTACs?
The selectivity of E3 substrate degradation can be evaluated from three dimensions:
- Whether the degradation of the natural substrate of E3 is affected.
- Known specific E3 neosubstrates.
- Other potential influencing factors.
References:
- Churcher, I. (2018). Protac-induced protein degradation in drug discovery: breaking the rules or just making new ones?. Journal of medicinal chemistry, 61(2), 444-452.
- Toure, M., & Crews, C. M. (2016). Small-molecule Protacs: new approaches to protein degradation. Angewandte Chemie International Edition, 55(6), 1966-1973.
* PROTAC® is a registered trademark of Arvinas Operations, Inc., and is used under license.
