E. coli is widely used to express proteins without post-translational modifications such as glycosylation to maintain activity, but a large number of E. coli expression often leads to the formation of protein aggregation and precipitation - inclusion bodies. Inclusion body proteins are without biological activity, requiring a complex solvent to dissolve, refolding and re-purification to obtain the biological active fractions. Typically, the use of high concentrations of denaturing agents (urea, guanidine hydrochloride) are necessary to dissolve inclusion bodies. To prevent denaturant refolding, reducing agents such as β-mercaptoethanol are added to remove the presence of oxidants.
This traditional method has a lot of shortcomings. Generally, the efficiency of refolded biologically active proteins is low - about 15-25%.

RealGene uses our patented technology for the refolding of inclusion bodies, and in this, greatly improved the success rate of protein re-naturation and recovery.

Service Advantages
For most proteins, we pledge that more than 95% of the inclusion bodies can dissolve and provide 85% of the final refolded purified products.
Realgene protein folding services include the following:
1. Physical and chemical analysis of the target protein
2. Optimization of inclusion body solvents.
The recovery strategy is based on the nature of protein sequence and structure. In general, our program includes the following:
Realgene patented refolding method
Direct dilution method
Dialysis and filtration
Refolding techniques: SEC (size exclusion chromatography), HIC (hydrophobic interaction chromatography), affinity chromatography, as well as IEC (ion exchange chromatography)
Delivery Information:
For 1 L expression system, we can provide customers with a purity of 85% of at least 5mg soluble protein complex. After recovery of soluble fraction, SDS-PAGE analysis will be carried out in accordance to customer’s required verification methods. Delivery will be in suitable buffer as according to customer specifications.