Expression of recombinant proteins in Escherichia coli often leads to formation of inclusion bodies (IBs). To recover the protein activity, the IBs are isolated, solubilized and refolded. The protein refolding processes play a major role in the production of recombinant proteins; thus, various methodologies have been implemented, including dilution, dialysis and column chromatography with or without the assistance of molecular chaperones. Recently, it was demonstrated that the apical domain of GroEL (AD), DsbA and DsbC immobilized on cellulose improved the efficiency of chromatographic refolding of rhodanese and lysozyme. Although immobilized chaperones and foldases greatly improve refolding yields, their use has been limited. To improve their potential use at the bioprocess scale, it is essential to understand the effects of operational conditions and additives on refolding yields. Therefore, we investigated the lysozyme refolding kinetics assisted by the apical domain of GroEL (AD), DsbA and DsbC in either soluble or immobilized on cellulose with different lysozyme concentrations, different chaperone:lysozyme ratios, absence of redox pairing, presence of glycerol and presence of high concentrations of GdnHCl and β−mercaptoethanol (β−ΜΕ). Our results provide insight to improve the use of molecular chaperones in the refolding of recombinant proteins expressed as inclusion bodies.