Background: Interferon α-2b is a vital biotherapeutic produced through the recombinant DNA technology in E. coli. The recombinant IFN-α2b normally appears as intercellular IBs, which requires intensive refolding and purification steps. Method: Purification of IFN-α2b from solubilized IB was performed using two-phase extraction. To optimize refolding conditions, the effects of pH and different a...

Low molecular size additives such as L-arginine and the redox compounds have been used both in the culturemedium and in vitro refolding to increase recombinant proteins production. Additives increase proteinrefolding and yield of active proteins by suppressing aggregate formation or enhancing refolding process.In this work, a comparative study was performed on refolding of rec...

We employed a urease-catalyzed reaction to gradually remove a high concentration of a chaotropic agent (urea) from a denatured protein solution and demonstrated that efficient protein refolding can be achieved by the urease-catalyzed reaction, without large-volume dilution.

Protein refolding is an important process to recover active recombinant proteins from inclusion bodies. Refolding by simple dilution, dialysis and on-column refolding methods are the most common techniques reported in the literature. However, the refolding process is time-consuming and laborious due to the variability of the behavior of each protein and requires a great deal of trial-and-error ...

background: the over-expression of recombinant proteins in large amount is important for production of therapeutic proteins and structural study. there are several systems for expression of recombinant proteins. one of the most relevant expression systems is escherichia coli (e. coli). although this organism has many advantages, most of recombinant proteins expressed in e. coli hosts form inclu...

A recurring obstacle for structural genomics is the expression of insoluble, aggregated proteins. In these cases, the use of alternative salvage strategies, like in vitro refolding, is hindered by the lack of a universal refolding method. To overcome this obstacle, fractional factorial screens have been introduced as a systematic and rapid method to identify refolding conditions. However, metho...

Over-expression of heterologous proteins in Escherichia coli is commonly hindered by the formation of inclusion bodies. Nevertheless, refolding of proteins in vitro has become an essential requirement in the development of structural genomics (proteomics) and as a means of recovering functional proteins from inclusion bodies. Many distinct methods for protein refolding are now in use. However, ...

The fast reaction ((T2) approximately 50 msec) observed previously in the refolding of thermally unfolded ribonuclease A (disulfide bonds intact) has now been studied by two properties indicative of enzyme function: binding of a competitive inhibitor (2'CMP) and hydrolysis of a substrate (CpA --> C > p + A). Both the binding and catalytic reactions are fast (<2 msec) compared to refolding. Bind...

L-arginine (Arg) is a widely used additive for suppressing protein aggregation during refolding. Systematic screening of Arg analogs provides superior additives that enhance the refolding yield more effectively than Arg. The refolding yield of hen egg lysozyme in the presence of 500 mM L-argininamide (ArgAd) increases 1.7-fold higher than Arg. Thermal unfolding experiments indicate that ArgAd h...

background: the refolding of proteins from inclusion bodies is affected by several factors, including solubilization of inclusion bodies by denaturants, removal of the denaturant, and assistance of refolding by small molecule additives. objectives: the purpose of this study was optimization of recombinant human interferon-b purification in order to achieve higher efficiency, yield, and a produc...