CD Biosynsis offers comprehensive E. coli Protein Expression and Purification Services , providing high-quality, functional proteins for research, structural biology, and assay development. As the most widely utilized host, E. coli provides a fast, cost-effective platform for recombinant protein production. Our service covers the entire workflow: from codon optimization and vector design to strain selection , expression optimization (including solubility enhancement), and multi-step chromatographic purification to achieve high purity (up to >98%) and high yield. We are experts in handling challenging proteins, including toxic, low-solubility, and complex multi-domain targets.
Get a QuoteWhile E. coli is a robust expression system, challenges such as the formation of insoluble inclusion bodies , poor yield of target protein, and issues with protein toxicity often arise. We mitigate these risks through a systematic optimization process. This involves careful selection of expression strains (e.g., BL21(DE3) variants for rare codons or specific proteases), tuning induction parameters (IPTG concentration, temperature), and applying co-expression strategies (chaperones) to maximize soluble and properly folded protein yield. Our purification process is tailored to the protein's properties to deliver the highest possible purity and activity.
Codon Optimization & Synthesis
Computational analysis and synthesis of DNA sequences optimized for E. coli translation machinery to boost expression levels.
Expression Vector Design
Cloning into various vectors with fusion tags (His-Tag, GST, MBP) and regulatable promoters (T7, T5) for controlled expression.
Solubility Screening
Systematic screening of E. coli strains, induction temperatures, and media types to maximize soluble protein fraction.
Affinity Chromatography
Primary purification step using affinity resins (e.g., Ni-NTA for His-Tagged proteins) followed by tag cleavage (optional).
Polishing Purification
Secondary purification using Ion-Exchange (IEX) and Size-Exclusion Chromatography (SEC) to achieve research-grade purity (>95%).
Inclusion Body Refolding
Expert protocols for solubilizing and properly refolding proteins isolated from insoluble inclusion bodies to restore biological activity.
Toxic Protein Production
Use of tightly regulated expression systems and specific host strains to minimize basal expression and cell death.
Multi-Subunit Assembly
Co-expression strategies for simultaneous production of multiple subunits (e.g., dimer, trimer) required for functional complexes.
Endotoxin Removal
Specialized purification protocols to remove LPS (endotoxin) down to clinical/cell culture grade levels (<1 EU/mg protein).
A sequential process ensuring maximum yield and verifiable purity.
Gene Optimization & Cloning
Expression Screening & Scale-Up
Purification & Tag Removal
Quality Control & Delivery
Design: Codon optimization and DNA synthesis for the target protein sequence.
Cloning: Insertion into appropriate expression vector with desired fusion tag and protease cleavage site.
Small-Scale Screening: Test various strains, temperatures, and media conditions for optimal soluble yield.
Scale-Up: Transition to large-scale fermentation (liters) for high-yield production based on optimized conditions.
Lysis & AC: Cell lysis followed by primary Affinity Chromatography.
Polishing: Optional protease cleavage of fusion tag, followed by IEX and SEC for high purity.
Guaranteed Purity and Yield
We offer flexible purity tiers (up to >98%) and high-yield production for milligrams to gram quantities.
Expert Solubility Optimization
Strategies including low-temperature induction, chaperone co-expression, and fusion tag screening to maximize soluble protein.
Low Endotoxin Processing
Specialized purification for sensitive applications, ensuring endotoxin levels are suitable for cell culture (<1 EU/mg) or in vivo studies.
Custom QC Assays
Beyond SDS-PAGE, we offer custom functional assays, Mass Spec verification, and aggregation analysis (SEC) upon request.
"We struggled to express our target enzyme due to toxicity. CD Biosynsis successfully optimized the system using a specialized E. coli strain and delivered 500 mg of active protein at >95% purity, which was crucial for our structural studies."
Dr. Emily Roth, PI, Structural Biology Lab
"The Low Endotoxin Purification service was outstanding. The final protein used in our cell therapy assays had undetectable LPS levels, saving us significant time and effort."
Mr. Kevin Zhou, R\&D Manager, Biotech Therapeutics
"They took our inclusion body project and expertly managed the Refolding process. The resulting protein was functionally active, something we couldn't achieve internally."
Ms. Lisa Nguyen, Research Scientist, Drug Discovery
"We commissioned CD Biosynsis to support an intricate gene editing project with multiple targets. Their talent in producing high-quality work in a short period of time was impressive. Their solutions were custom made to suit our needs, and they went above and beyond to ensure our experiments worked. Their support has been a great asset to our research department and we look forward to further working with them."
Dr. Raj Patel, Principal Investigator, Department of Molecular Biology
"As a pharmaceutical company working to discover new cancer therapies, we require accurate, trustworthy gene editing solutions. CD Biosynsis did more than what we expected when it came to providing strong, accurate CRISPR/Cas9 solutions for our preclinical research. Their technical support team was excellent and responsive, and they quickly replied to our questions. This alliance has been pivotal in helping us move our drug pipeline forward. Thank you, CD Biosynsis, for your amazing service!"
Dr. Clara Rodriguez, Chief Scientist, AstraZeneca Pharmaceuticals, Spain
What is the typical purity level guaranteed?
We typically guarantee a minimum purity of >95% verified by SDS-PAGE. For highly sensitive applications (e.g., crystallization), we offer an ultra-high purity grade of >98% verified by HPLC or SEC.
How do you handle proteins that form inclusion bodies?
If initial screening results in inclusion body formation, we isolate the inclusion bodies, solubilize the protein using denaturants, and apply carefully controlled Refolding protocols to restore the native, functional structure of the protein before final purification.
What is the significance of Endotoxin Removal?
Endotoxin (LPS) is a cell wall component of E. coli that is highly inflammatory. Its removal is critical if the protein will be used in cell culture, in vivo studies, or clinical applications, where LPS levels must be reduced to <1 EU/mg protein.
Do you offer a functional assay for the final product?
Yes, while our standard QC includes SDS-PAGE and concentration, we offer custom functional QC tests, such as ELISA, enzyme kinetic assays, or SPR/BLI binding assays, upon client request to confirm biological activity.
How much does Metabolic Engineering services cost?
The cost of Metabolic Engineering services depends on the project scope, complexity of the target compound, the host organism chosen, and the required yield optimization. We provide customized quotes after a detailed discussion of your specific research objectives.
Do your engineered strains meet regulatory standards?
We adhere to high quality control standards in all strain construction and optimization processes. While we do not handle final regulatory approval, our detailed documentation and compliance with best laboratory practices ensure your engineered strains are prepared for necessary regulatory filings (e.g., GRAS, FDA).
What to look for when selecting the best gene editing service?
We provide various gene editing services such as CRISPR-sgRNA library generation, stable transformation cell line generation, gene knockout cell line generation, and gene point mutation cell line generation. Users are free to select the type of service that suits their research.
Does gene editing allow customisability?
Yes, we offer very customised gene editing solutions such as AAV vector capsid directed evolution, mRNA vector gene delivery, library creation, promoter evolution and screening, etc.
What is the process for keeping data private and confidential?
We adhere to the data privacy policy completely, and all customer data and experimental data are kept confidential.
CRISPR-Cas9 technology represents a transformative advancement in gene editing techniques. The main function of the system is to precisely cut DNA sequences by combining guide RNA (gRNA) with the Cas9 protein. This technology became a mainstream genome editing tool quickly after its 2012 introduction because of its efficient, simple and low-cost nature.
The CRISPR gene editing system with its Cas9 version stands as a vital instrument for current biological research. CRISPR technology enables gene knockout (KO) through permanent gene expression blockage achieved by sequence disruption. Various scientific domains including disease modeling and drug screening employ this technology to study gene functions. CRISPR KO technology demonstrates high efficiency and precision but requires confirmation and verification post-implementation because unsatisfactory editing may produce off-target effects or incomplete gene knockouts which impact experimental result reliability. For precise and efficient Gene Editing Services - CD Biosynsis, Biosynsis offers comprehensive solutions tailored to your research needs.
The CRISPR-Cas9 knockout cell line was developed using CRISPR/Cas9 gene editing to allow scientists to remove genes accurately for research on gene function and disease models and pharmaceutical discovery. Genetic research considers this technology essential due to its high efficiency together with simple operation and broad usability.
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CD Biosynsis is a leading customer-focused biotechnology company dedicated to providing high-quality products, comprehensive service packages, and tailored solutions to support and facilitate the applications of synthetic biology in a wide range of areas.