DNA Damage Response (DDR) Protein Engineering Service
IntroductionStrategiesWhat We Can OfferWorkflowCustomer ReviewsFAQsExtended Services
DNA Damage Response (DDR) Protein Engineering Service: Accelerate Your Drug Discovery Process!
The DDR is a sophisticated cellular network crucial for maintaining genomic integrity. It detects DNA lesions, signals their presence, and orchestrates repair pathways, protecting against mutagenesis and disease. Key players include sensor proteins (e.g., PARP1, Ku70/80), transducers (e.g., ATM, ATR, DNA-PKcs, Chk1/2), and effectors. Dysregulation of DDR pathways is a hallmark of cancer, making DDR proteins prime targets for therapeutic intervention. Engineering high-quality, functional recombinant DDR proteins is paramount for biochemical, structural, and drug discovery research.
DDR protein engineering service at creative Biolabs helps you accelerate drug discovery and obtain high-quality recombinant proteins through advanced protein expression, purification, and modification techniques, ensuring their functional integrity for your critical studies. This service is meticulously designed to provide researchers and pharmaceutical companies with precisely engineered DDR proteins essential for advancing their projects. We deliver highly purified, functionally active recombinant DDR proteins, including kinases (e.g., ATM, ATR, Chk1, DNA-PKcs) and PARP enzymes, critical for drug screening, structural biology, and mechanistic studies. Our service directly addresses challenges such as low expression yields, solubility issues, and the need for specific post-translational modifications, enabling you to accelerate target validation, inhibitor development, and the comprehensive understanding of DDR pathways.
Strategies of This Service
Our approach to DDR Protein Engineering is founded on a multi-faceted strategy designed to overcome common challenges associated with expressing and purifying complex eukaryotic proteins. This includes:
Optimized Expression System Selection: We meticulously select the most appropriate expression system (e.g., E. coli, baculovirus/insect cells, mammalian cells) based on the specific DDR protein's characteristics, size, required post-translational modifications (e.g., phosphorylation, ubiquitination), and desired functional activity. This ensures optimal folding and PTMs critical for DDR protein function.
Rational Construct Design: Our experts employ rational design principles for gene constructs, incorporating various tags (e.g., His, GST, MBP) for enhanced solubility and purification efficiency. We also consider truncations or domain-specific expression where full-length proteins are challenging, focusing on functionally relevant regions.
Codon Optimization: For heterologous expression, we perform codon optimization tailored to the chosen expression host, significantly improving protein synthesis rates and yields, particularly for AT-rich or GC-rich genes encoding DDR proteins.
Advanced Purification Techniques: We utilize a comprehensive suite of chromatography methods, including affinity chromatography, ion exchange chromatography, and size-exclusion chromatography (SEC). SEC is particularly crucial for ensuring the proper oligomeric state and homogeneity of DDR proteins, which often function as complexes.
Rigorous Quality Control & Functional Validation: Beyond standard purity assessments (SDS-PAGE, Western Blot), we employ specific biochemical and biophysical assays to confirm the functional activity of DDR proteins. This includes enzyme activity assays (e.g., FRET-based kinase assays for ATM/ATR/Chk1, PARP activity assays), binding assays, and thermal stability assays to ensure stability and functionality for downstream applications.
Custom Post-Translational Modification (PTM) Integration: Recognizing the critical role of PTMs in DDR protein function, we offer strategies for incorporating specific modifications, such as phosphorylation, either through co-expression with relevant kinases or in vitro enzymatic modifications, providing functionally accurate proteins.
What We Can Offer
Creative Biolabs offers a comprehensive suite of services under our DDR Protein Engineering umbrella, specifically tailored to meet the diverse needs of academic and industrial research:
Custom Recombinant DDR Protein Production: We provide high-quality, recombinant production of a wide range of DDR proteins, including but not limited to:
Kinases: ATM, ATR, DNA-PKcs, Chk1, Chk2.
PARPs: PARP1, PARP2.
Repair Enzymes: Ligases (e.g., DNA Ligase IV), Nucleases (e.g., MRE11).
Scaffold Proteins: BRCA1, 53BP1. We ensure optimal expression, purification, and functional validation for each protein.
Protein Engineering for Specific Needs:
Mutagenesis Services: Introduction of specific point mutations, deletions, or insertions to study protein function, structure-activity relationships, or drug resistance mechanisms within DDR proteins.
Domain-Specific Expression: Production of individual domains or truncated versions of large DDR proteins for focused structural and functional studies.
Tagged Protein Production: Generation of DDR proteins with various fusion tags (e.g., His-tag, GST-tag, MBP, SUMO) to enhance solubility, aid purification, and facilitate detection.
Functional Characterization and Quality Control:
Enzyme Activity Assays: Comprehensive biochemical assays to confirm the enzymatic activity of purified DDR proteins (e.g., kinase activity assays, PARP activity assays, ATPase assays).
Biophysical Characterization: Assessment of protein stability (e.g., by DSF/nanoDSF), oligomeric state (e.g., by SEC-MALS), and binding kinetics (e.g., by SPR/BLI) to ensure suitability for structural and mechanistic studies.
Mass Spectrometry for PTM Analysis: Verification and characterization of crucial post-translational modifications (e.g., phosphorylation, ubiquitination) on DDR proteins.
Scale-Up Production: From milligram to gram quantities, we can scale up production to meet the demands of large-scale screening campaigns or preclinical studies.
Workflow
Required Starting Materials: To initiate your project, we typically require:
The gene sequence (cDNA or gBlock) of your target DDR protein.
Information on any desired tags (e.g., His-tag, GST-tag) or specific mutations.
Details on post-translational modifications (e.g., phosphorylation sites) if specific modifications are required for functional studies.
Any specific functional assay requirements or target purity levels?
Customer Reviews
[Exceptional Quality] "Using DDR protein engineering service at Creative Biolabs in our research has significantly improved our ability to perform consistent and reliable kinase inhibition assays. The ATM protein provided was of unprecedented purity and activity, streamlining our lead optimization efforts for a new class of DDR-targeted therapeutics." 2024-03-15, Dr. Li*****g.
[Reliable & Efficient] "Expertise in engineering challenging proteins, specifically their DNA-PKcs, was a game-changer for our structural biology project. We had struggled with expression and purification for months, but their service delivered a highly soluble and stable protein, enabling us to obtain high-resolution cryo-EM data. Their attention to detail regarding specific modifications made all the difference." 2023-11-20, Prof. An****a.
[Tailored Solutions] "The team at Creative Biolabs provided an incredibly tailored solution for our need for a specific mutant ATR protein. Their ability to deliver a functionally validated protein for our cell-based assays, addressing potential off-target effects through their purification process, was invaluable. This service truly stands out compared to other providers, offering superior functional protein for complex research." 2024-01-05, Dr. Ch*****n.
FAQs
What types of DDR proteins can Creative Biolabs engineer?
We have extensive experience with a broad spectrum of DDR proteins, including key kinases like ATM, ATR, Chk1, and DNA-PKcs, as well as PARP enzymes, and various repair complex components. If your specific protein isn't listed, please reach out!
How do you ensure the functional activity of the engineered DDR proteins?
Beyond standard purity assessments, we employ rigorous functional validation, including specific enzymatic assays (e.g., kinase activity, PARP activity), biophysical characterization, and mass spectrometry, to guarantee that the delivered protein is not only pure but also functionally active for your research.
How do you address solubility or expression challenges with difficult DDR proteins?
Our expertise lies in overcoming these challenges. We employ a multi-pronged approach including rational construct design, codon optimization, screening of various expression systems and conditions, and the use of solubility-enhancing tags to ensure successful protein production.
Can I request a specific mutation or truncation for my DDR protein of interest?
Yes, our service is highly customizable. We can introduce specific point mutations, deletions, or express truncated domains of your DDR protein to facilitate structure-function studies or explore specific interactions.
Is there a minimum or maximum quantity of protein I can order?
We are flexible and can accommodate various scales, from milligram quantities for initial studies to gram-scale production for large-scale screening or preclinical applications. Please discuss your quantity needs during the consultation.
How does your service compare to in-house protein production?
Our service offers significant advantages by leveraging specialized expertise, advanced equipment, and established workflows, often leading to higher yields, better purity, and faster turnaround times, freeing up your valuable in-house resources.
What if I need assistance with experimental design related to my DDR protein?
Our experienced scientific team is available for consultation throughout your project. We can provide guidance on experimental design, help interpret results, and offer troubleshooting support to ensure your success.
Extended Services
Creative Biolabs offers a suite of complementary services designed to provide a comprehensive solution for your drug discovery and research needs, extending beyond our core DDR Protein Engineering. These services seamlessly integrate to support your project lifecycle from target identification to lead optimization.
PARP Inhibitor Screening Services: Discover potent PARP inhibitors using our functionally validated recombinant PARP enzymes and diverse assay formats, accelerating your oncology drug development.
Structural Biology Services: Utilize our highly purified DDR proteins for structural studies (e.g., X-ray crystallography, cryo-EM), providing critical insights into protein-ligand interactions and rational drug design.
Custom Antibody Development: If you require specific antibodies against DDR proteins for research or diagnostic purposes, our custom antibody development service can generate high-affinity, specific antibodies.
Cell-Based Assay Development: For functional studies of DDR pathways in a cellular context, we offer services to develop and optimize cell-based assays using our engineered proteins.
At Creative Biolabs, DDR protein engineering service is your trusted partner for obtaining high-quality, functionally validated recombinant DDR proteins. Our deep scientific expertise, advanced technologies, and commitment to client success ensure that your projects are accelerated, from basic research to preclinical drug discovery. We are dedicated to providing the precise tools you need to unravel complex DDR mechanisms and develop novel therapeutic interventions.
Ready to advance your research with high-quality DDR proteins? Contact our expert team today to discuss your specific project requirements, receive a personalized quote, and learn how Creative Biolabs can be instrumental in your success.
