Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process

Table 2 Methods used for refolding of solubilized inclusion body proteins

Refolding methods	Remarks	Reference
Dilution
Flash dilution	Simplest method	[59]
Pulsatile dilution	low requirement of buffer and improved refolding yield	[54]
Dialysis
One step dialysis	May be successful only for those proteins that are soluble in intermediate states	[74]
Step wise dialysis	Useful for multidomain or disulphide bond containing proteins	[74]
On column refolding
Size exclusion chromatography	Separation of folded form from intermediates	[75-79]
Anion exchange chromatography	More advantageous for crude samples	[80-83]
Affinity chromatography	Limited to cases where the Tag doesn’t interfere with folding	[84-86]
Hydrophobic interaction chromatography	May substitute for the requirement of additives during refolding	[87,88]
Chromatography in presence of chaperones	Reduces aggregation by mimicking in vivo scenario	[91-95]
Micro fluidic chips	May be useful for difficult to fold proteins	[98]
Urease mediated refolding	No requirement of refolding buffer	[99]

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