Protein Refolding Services
Insoluble aggregates, also called inclusion bodies, are one of the major issues facing protein expression in E. coli. The insoluble aggregates are non-functional but completely block the progress of research into downstream applications. GenScript's protein refolding services provide a solution to this problem.
Our confidence is backed by our proprietary refolding technology, which includes high-hydrostatic-pressure techniques, small-molecule additives, and column-refolding methods.
- Service Specifications
- Case Studies
- Related Services
Protein refolding services are provided along with our Bacteria Protein Service Packages upon request. Over 95% of the inclusion bodies can be solublized and refolded. We provide refolded protein at purity of 85% or greater. The final output is dependent on the total amount of target protein comprising the aggregates. The refolded protein is delivered in customized buffer. The major elements of our protein refolding services include the following procedures:
- Evaluation of target proteins' biochemical and biophysical properties
- Refolding optimizations in the 1 L system
- The particular refolding strategy is selected based on protein's sequence and the structural properties. Typically, our strategies include the following:
- Direct dilution
- Dialysis and diafiltration
- Small molecular assistants:
- Amino acids
- Polymers: micellar and liposomal
- Reducing/oxidizing reagents
- Salts: Na/K/Mg-Cl and (NH4)2SO4
- Saccharides: glucose, sucrose, and trehalose
- Artificial chaperones: detergent and non-detergent
- Matrix-assisted methods: SEC (size exclusion chromatography), HIC (hydrophobic interaction chromatography), affinity column and IEC (ion exchange chromatography)
- Chaperon-mediated methods: ATP-dependent and ATP-independent
- Gradient methods: pH, Tem, hydrostatic pressure, and denaturant
1. A Protein denaturation and renaturation from inclusion bodies:
|Fig. A-1 (left figure): Inclusion body preparation and denaturation |
Lane 1: Total protein after sonication
Lane 2: The supernatant after centrifugation
Lane 3: Washed with 1M urea
Lane 4: Dissolved by 8M urea
|Fig. A-2 (right figure): Refolded protein |
Lane 1: Refolded tag-free A protein
2. B Fusion protein denaturation and renaturation from inclusion bodies:
|Fig. B-1 (left figure): Inclusion body preparation and denaturation |
Lane 1: The supernatant after centrifugation
Lane 2: The precipitation after centrifugation
Lane 3: Inclusion bodies dissolved by urea
Lane 4: Flow through
Lane 5: Washed by imidazole containing urea
Lane 6: Eluted by imidazole containing urea
|Fig. B-2 (right figure): Refolded protein |
Lane 1 and 2: Refolded B fusion protein
1. OptimumGeneTM Codon Optimization Technology: Our proprietary OptimumGeneTM codon optimization technology comprehensively optimizes factors critical to transcription, translation, and co-translational protein folding to deliver the highest possible levels of expression in any system.
Minimum 10-fold increase in protein express level
2. Bacterial Expression: GenScript has developed BacPowerTM technologies to tackle hard-to-express and hard-to-dissolve proteins. Guaranteed package (SC1253): 3 mg purified protein.
3. Yeast Expression: Our yeast expression system employs stable and durable production strains that resemble mammalian protein processing, incorporating features of high yield, high productivity, and dramatic reductions in costs and time requirements.
4. Insect Cell/Baculovirus Expression: GenScript has pioneered methods for the secretion of recombinant proteins from baculovirus-infected insect cells. Our services are suitable for academic institutions and both the pharmaceutical and biotechnology industries.
5. Mammalian Cell Expression: Our proprietary 293 and CHO suspension system platform promotes fast production and delivery of recombinant proteins and monoclonal antibodies up to grams level.
6. Stable Cell Line Develipment: Custom stable cell lines for assay development, recombinant protein and monoclonal antibody production. We also provide comprehensive assay-ready cell line productions to preserve your time for higher value projects.
GenScript can provide soluble refolded protein at the purity of 85% or greater from one-liter expression system, based on total target protein comprising the aggregates. The refolded protein is delivered in customized buffer with the protein purity and identity data determined by Coomassie-stained gels and western blot analyses using Tag or the customer's designated protein-specific antibody (must be provided by the customer).
Quotations and Ordering:
To request a quotation, please download and complete the Quotation/Order Form, and send it to us by email or fax.
To order, please download and complete the Quotation/Order Form and send it to us by email or fax with a formal PO (Purchase Order) or credit card information. You can also submit PO/credit card information by phone or via our secure online messaging system.
If submitting samples, please mail them together with a hard copy of the completed Quotation/Order Form to: Protein Services, GenScript, 860 Centennial Ave., Piscataway, NJ 08854, US.
For questions, or to inquire about the status of your order contact us by email, phone, fax or via our secure online messaging system.
Our customer service representatives are available 24 hours a day, Monday through Friday to assist you.
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