Methods of renaturation of recombinant proteins
AssigneeAerovance, Inc.
InventorsWayne Froland, Katherine Delaria, David Boisvert, Robert Kuhn
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Abstract
The present invention provides large-scale methods for renaturation of proteins comprising adding a solution of denatured, chemically modified or reduced proteins to a refolding buffer containing sulfate derived from H2SO4 and/or MgSO4 in the presence of guanidine. The present invention further provides methods of isolating a refolded protein at a concentration of about 0.4 to 3.0 gm/L by using a hydrophobic interaction chromatography (HIC) column.
Claims43
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A method of isolating a refolded protein at a concentration of about 0.4 to 3 gm/L comprising:
- a) loading a pH-adjusted protein solution to a hydrophobic interaction chromatography (HIC) column;
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- b) eluting the protein with an elution buffer.
A method for large-scale renaturation of proteins comprising adding to a refolding buffer a protein solution of about 12 to 20 gm/L of denatured, chemically modified or reduced protein in the presence of guanidine, wherein the refolding buffer comprises MgSO4 in a Tris-base/Tris-HCl system.
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A method for large-scale renaturation of proteins comprising:
- a) diafiltering a denatured, chemically modified, or reduced protein with 1 to 10 diavolumes of a diafiltration buffer, wherein the diafiltration buffer comprises guanidine;
- b) concentrating the diafiltered protein to about 12 to 20 gm/L;
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- c) adding the concentrated protein via pulsed dilution to a refolding buffer, wherein the refolding buffer comprises MgSO4 in a Tris-base/Tris-HCl system.
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Dependencies are inferred from the claim text.