High pressure fosters protein refolding from aggregates at high concentrations.
High hydrostatic pressures (1-2 kbar), combined with low, nondenaturing concentrations of guanidine hydrochloride (GdmHCl) foster disaggregation and refolding 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 suspensions containing aggregates of recombinant human growth hormone (up to 8.7 mg/ml) and 0.75 M GdmHCl. Covalently crosslinked, insoluble aggregates of lysozyme could be refolded to native, functional protein at a 70% yield, independent of protein concentration up to 2 mg/ml. Inclusion bodies containing beta-lactamase could be refolded at high yields of active protein, even without added GdmHCl.
St John
R J
RJ
Department of Chemical Engineering, University of Colorado, Boulder, CO 80309, USA.
Carpenter
J F
JF
Randolph
T W
TW
D013006:Growth Hormone / Q000737:chemistry*; D019791:Guanidine / Q000737:chemistry; D006801:Humans; D009113:Muramidase / Q000737:chemistry*; D011312:Pressure; D011489:Protein Denaturation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry; D001618:beta-Lactamases / Q000737:chemistry*
D011994:Recombinant Proteins; D013006:Growth Hormone; D009113:Muramidase; D001618:beta-Lactamases; D019791:Guanidine