Protein Expression and Strain Development
We offer early insights into the expression of your protein, delivering purified proteins for research use in as little as 5 weeks. With extensive experience covering various bacterial expression systems, including a profound understanding of E. coli and L. lactis systems, we stand ready to assist you in selecting the optimal microbial strain tailored to your specific requirements.
Our comprehensive approach includes assessing expression levels, solubility and cellular localization, providing valuable insights into the manufacturability of your protein and facilitating informed decision-making from the project’s inception. Backed by a proven track record of developing fully documented tech-transfer ready processes that can be transferred to established CMOs, we lay the groundwork to propel your project towards manufacturing success.
- Systems: E. coli, L. lactis, Your preferred system
- Test multiple constructs and expression conditions
- As little as 5 weeks to purified protein
- Gram Yields
- Development of Research and Master Cell Banks
Workflow at Bioneer
Leveraging our expertise across diverse protein types, we provide early insights into the expression of your protein and mitigate risk through a flexible step-wise project approach with Go/No-Go decision points.
Process Step |
Our approach |
Time |
|---|---|---|
|
Gene Design and Plasmid Preparation
|
|
2 – 4 weeks
|
| Strain Selection and Banking |
|
2+ weeks
|
| Feasibility Studies |
|
2 – 3 weeks
|
Report Outlining Expression Results (Go/No Go) |
||
| Upscaled Expression in Shake-Flasks |
|
2 – 3 weeks
|
| Batch and Fed-batch Production & Upstream Process Development |
|
2+ weeks
|
Reporting & Delivery of Culture Supernatant/Cell Pellets/Inclusion Bodies (Go/No-Go) |
||
| Purification and Downstream Process Development |
|
2+ Weeks
|
|
Characterization and Analysis
|
|
2+ Weeks
|
|
Formulation
|
|
2+ Weeks
|
Reporting and Delivery of Purified Protein |
Additional Services:
Our Process and Approach
Report Outlining Expression Results (Go/No Go)
Reporting & Delivery of Culture Supernatant/Cell Pellets/Inclusion Bodies (Go/No-Go)
Reporting and Delivery of Purified Protein
Additional Services:
Upstream Process Development
We provide comprehensive assistance in upstream process development for microbial systems, covering batch and fed-batch processes, preparing them for manufacturing.
Our expertise spans from conceptualization to scalable implementation, ensuring a smooth transition from laboratory to larger production volumes. By meticulously tailoring cultivation conditions, we optimize protein expression in your microbial strain. With the capability to scale up to 50 L in steel fermentors, we offer a robust platform for process validation and production scale-up. Whether you’re initiating process development, scaling up production, or conducting validation studies, our dedicated and experienced team is committed to supporting your upstream bioprocessing needs with efficiency and precision.
- Standard Batch and Fed-batch Processes
- Customized Processes
- 6x 1 L glass fermentors
- 1x 15 L steel fermentor
- 4x 50 L dedicated steel fermentors for manufacturing
- Periplasmic and cytoplasmic extraction
- Inclusion Body preparation
Purification and Downstream Process Development
We specialize in developing customized downstream processes, drawing upon our profound understanding of the inherent complexities in protein purification and the unique characteristics of each protein.
We can rapidly provide materials using standardized methods such as affinity chromatography or devise intricate multistep processes that ensure purity levels surpassing 99%. With experience in developing processes for cGMP manufacturing, our processes are carefully designed for seamless transfer to manufacturing facilities. Whether you require rapid material, full-scale process development, or optimization of existing procedures, our team of dedicated experts is fully equipped to assist you every step of the way.
- Tagged and Non-tagged Methods
- Upscaling and Tangential Flow Filtration
- Process Development & Troubleshooting
- mg to g scale
- Inclusion Body Solubilization & Refolding
Variety of chromatographic techniques available
- Affinity chromatography (Protein A, L, IMAC, etc.)
- Ion exchange chromatography
- Size exclusion chromatography
- Mixed mode chromatography
- Hydrophobic interaction chromatography
- Endotoxin removal chromatography
- More!
Workflow at Bioneer
Leveraging our expertise across diverse protein types, we provide early insights into the expression of your protein and mitigate risk through a flexible step-wise project approach with Go/No-Go decision points.
Process Step |
Example techniques |
|
|---|---|---|
| Preparation, Extraction, Clarification |
|
|
|
Capture (Isolate, concentrate, and stabilize) |
|
|
| Intermediate Purification
(Remove Bulk Impurities) |
|
|
| Polishing
(Achieve final high level purity) |
|
|
| Finalizing/ Formulation
(final preparation) |
|
Our Process and Example techniques
Characterization & Analysis
We specialize in offering customizable characterization packages for protein analysis, utilizing a diverse array of techniques to meticulously examine protein properties. Through close collaboration with you, we tailor a plan to explore various facets of your protein, including purity, identity, structure, aggregation and stability.
Additionally, our cross-disciplinary approach enables us to provide further insights into your protein’s functionality by leveraging our in vitro modelling, histology, or formulation services. Whether you’re immersed in fundamental research, assay production, or biopharmaceutical development, our comprehensive characterization packages deliver invaluable insights to propel your projects forward.
Select from a variety of techniques
- Gel electrophoresis (SDS-PAGE, native PAGE, IEF)
- Western blotting
- Capillary electrophoresis (CGE and cIEF)
- Bradford and BCA protein quantification
- Quantitative amino acid analysis (outsourced)
- UV spectrophotometry
- UHPLC (SEC, IEX, HIC, RP chromatography)
- ELISA
- Enzyme assays
- Endotoxin measurement
- Host cell protein analysis (CHO and E. coli HCP, Cygnus kit)
- Host cell DNA (qPCR method)
- Mass spectrometry (outsourced)
- Circular Dichroism
- Dynamic light scattering
- Infrared spectroscopy
- Thioflavin T Fluorescence
- DSC and ITC
- Mass Photometry (Refeyn TwoMP)
Additional Services:
For further information
please contact:
Ryan Jeffrey Polito
Business Development Manager
