High pressure fosters protein refolding from aggregates at high concentrations

High hydrostatic pressures (1-2 kbar), combined with low, nondenaturing con centrations of guanidine hydrochloride (GdmHCl) foster disaggregation and r efolding of denatured and aggregated human growth hormone and lysozyme, and beta-lactamase inclusion bodies. One hundred percent recovery of properly folded protein can be obtained by applying pressures of 2 kbar to suspensio ns containing aggregates of recombinant human growth hormone (up to 8.7 mg/ ml) and 0.75 M GdmHCl. Covalently crosslinked, insoluble aggregates of lyso zyme could be refolded to native, functional protein at a 70% yield, indepe ndent of protein concentration up to 2 mg/ml, Inclusion bodies containing b eta-lactamase could be refolded at high yields of active protein, even with out added GdmHCl.