Protein folding refolding service

Protein folding refolding service

E. coli is widely used to express proteins that do not require post-translational modifications such as glycosylation to maintain activity. However, large amounts of E. coli expression of recombinant proteins often cause them to aggregate and form protein precipitates-inclusion bodies. Inclusion body proteins have no biological activity and require complex dissolution, renaturation, and purification to obtain functionally active products. Generally, a high concentration of denaturant (urea, guanidine hydrochloride) is used to dissolve the inclusion body, and a reducing agent such as β-mercaptoethanol is added. After the protein is dissolved, the denaturant is renatured by slowly removing the denaturant in the presence of an oxidant.

Disadvantages of the traditional renaturation method of inclusion bodies: (1) Dissolving the inclusion body with a high concentration of denaturant will cause the loss of secondary structure of the protein, which will randomly wind the peptide chains and expose the hydrophobic surface. The main reason for the low efficiency of active proteins is that they lose the secondary structure of the protein during dissolution, and the denatured proteins are intertwined with each other during the renaturation process to aggregate and precipitate. (2) In many cases, the recovery rate of inclusion body renaturation is about 15-25%, which accounts for most of the cost of recombinant protein production in E. coli.

RealGene uses the company's patented renaturation technology to simplify protein renaturation procedures, greatly improving the success rate and recovery rate of protein renaturation.

service advantage

If you need, we guarantee that more than 95% of the inclusion bodies can be dissolved and folded and renatured. The purity of the renatured protein provided to customers is more than 85% (the final purity depends on the number of target proteins containing the inclusion bodies), and stored In a specific buffer.
The main contents of the refolding and refolding service of Rage Protein are as follows:
1. Analyze the physical and chemical properties of the target protein
2. Explore the gentle dissolution method of inclusion body protein.

The renaturation strategy is determined according to the sequence and structural properties of the protein. In general, our plan includes the following:
Ruiji's patented renaturation method: direct dilution dialysis and diafiltration renaturation techniques: SEC (volume exclusion chromatography), HIC (hydrophobic interaction chromatography), affinity columns, and IEC (ion exchange chromatography)

Delivery information:
In the 1L expression system, we can provide customers with 85% purity and 5mg soluble refoldable protein. The soluble renatured protein is analyzed by SDS-PAGE, verified by the liveness method provided by the client, and the work report is provided. The protein is delivered to the client in a specific buffer.