Despite the development of the eukaryotic expression systems, E. coli remains the most widely used expression system for recombinant proteins. Advantages of E. coli expression are low cost, high expression level, easy to scale up, short turnaround time. One point to keep in mind is that expression of foreign proteins in E. coli may result in the formation of protein aggregates, commonly known as inclusion bodies, as a result of imbalance between protein aggregation and solubilization. In this case, our protein refolding system are set to solubilize and refold the inclusion bodies and thereby obtain soluble, bioactive recombinant proteins.
Novoprotein’s bacteria expression system serves you with these advantages:
Collection of more than 10 host strains and various proprietary or non-proprietary vectors guarantees the selection of the most efficient expression system.
Enhanced fusion tag technology and proprietary AIE technology are available in house to increase protein solubility and production, and to decrease purification cost, even for toxic proteins.
We have established and optimized over the years a sophisticated protein refolding system composed of 20 refolding buffers to increase the chance of obtaining soluble and bioactive proteins.
We are able to produce up to 500L of E. coli cell culture for gram scale production.
Metabolic labeling with stable isotopes (13C and 15N) and biotinylation are available.
5mg protein at 80% purity from only $1000!
Timeframe and pricing
- Your gene of interest
- Evaluate and choose the suitable expression system
- Codon Optimization
- Sub-clone into E. coli Expression Vectors
- Transfect expression plasmid into E. coli
- Select high-expression transformants
- Expression evaluation and optimization
- Protein Refolding
- Large scale expression
(up to 500L,gram scale)
- SDS Page Gel
- Affinity column
- Ion exchange chromatography
- Hydrophobic chomatography
- Size exclusion chromatography
- Ready to go!
|Gene Cloning and Expression Test|
|Strategy design||Free||1 day|
|Codon optimization||Free||2-3 weeks|
|Subcloning gene of interest into expression vector||$195.00|
|Expression strain construction and expression optimization||$100.00||2 weeks|
|Pilot Scale Expression and Purification|
|2L bacterial cell culture||From $1,000.00||2-4 weeks|
|5mg of protein (soluble, >80% purity)|
|Large Scale Production|
|Large-scale fermentation up to 500L with 60-80g cell/L||From $1,000.00||1-2 weeks|
|Purification to gram-scale protein|
|Protein production to kilogram scale||Please inquire|
- Secreted cytokine
- Expressed as pro-peptide (33kD), processed to mature form (14kD)
Both pro-peptide and mature forms are expressed in inclusion body (IB)
- Pro-peptide modification to facilitate the refolding
- Solubilize and refold pro-peptide
- Remove pro-peptide sequence
Refolded bioactive mature peptide above 95% purity
1.Expression strain screeningFig.1 SDS-PAGE analysis of pro-peptide expression.
Lane2: After inducing
Lane3: Before inducing
2.Inclusion body solubilization and pro-peptide refoldingFig.2 Solubilization and refolding of inclusion body in NP proprietary buffers.
IB: Inclusion body
R: Soluble and folded pro-peptide, Reduced
NR: Soluble and folded pro-peptide, Non-Reduced
3.Purification of refolded pro-peptide
Fig.3 IEX purification (SP-HP) of refolded pro-peptide
FT: Flow Through
4.Mature peptide processingFig.4 Pro-peptide processed into mature form with NP proprietary reagents and buffer.
mp(R): Mature peptide (reduced)
mp(NR): Mature peptide (non-reduced)
5.Final product: refolded bioactive mature peptide with purity above 95% (Mass Spec confirmed)Fig.5 Final product analyzed by SDS-PAGE.
For more information about our E. coli expression service, contact us at firstname.lastname@example.org or visit our Service page. In addition, we also offer other protein expression services including yeast protein expression services, baculovirus expression services,mammalian expression service. To know how other scientists speak of us, check out our References.
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I feel like losing control of data generation!
Our service team keeps tight monitoring and tracking over each step where data are generated.