An analysis of the factors that affect the dissociation of inclusion bodies and the refolding of endostatin under high pressure

Efficient refolding of the bifunctional therapeutic fusion protein VAS-TRAIL by a triple agent solution

“…3). The protein solubilized by the triple agent solution containing 2 M urea showed an emission maximum of around 340 nm, indicating that the tryptophan residues of the solubilized VAS-TRAIL exist in a more hydrophobic environment [9]. With the further increase in urea concentration (from 2 M to 6 M), the solubilized VAS-TRAIL protein showed a red shift in emission maximum, indicating the loss of secondary structures.…”

“…This modification improves the overall recovery of bioactive proteins from IBs. Several mild solubilization techniques have been developed, including the use of high hydrostatic pressure [9], detergents, such as n-lauroyl-L-glutamate [10], organic solvents, such as n-propanol [11], and additives, such as L-arginine [12,13].…”

Investigation on solubilization protocols in the refolding of the thioredoxin TsnC from Xylella fastidiosa by high hydrostatic pressure approach

“…However, there are also articles in the literature that indicate that protein folding occurs during incubation at lower pressure levels (0.3-0.7 kbar) [13,27] or at atmospheric pressure [9]. To determine the conditions that favor TsnC refolding, IB suspensions were subjected to incubation at 2.4 kbar for 16 h. As alternative conditions, the IB suspensions were compressed at 2.4 kbar for 90 min for dissociation of the aggregates, which was followed by decompression to 0.8, 0.4, 0.2 kbar or 1 bar for refolding, condition that was maintained for 16 h before complete decompression and dialysis.…”

Solubilization of inclusion body proteins using low and very low concentrations of chemicals: implications of novel combined chemical treatment designs in enhancement of post‐solubilization target protein purity and biological activity

“…High hydrostatic pressure (2–4 kbar), alone or combined with guanidinium chloride, has also been introduced as a mild method for solubilization of proteins from inclusion bodies produced in E. coli . While high pressure disrupts the intermolecular interactions and disaggregates inclusion bodies, it maintains the secondary structure of the solubilized proteins and protein refolding is facilitated following removal of the applied pressure …”