Process for renaturation of recombinant, disulfide containing proteins at high protein concentrations in the presence of amines
View Patent ↗A method for renaturation of proteins comprising adding to a solution of denatured, chemically modified or reduced proteins a refolding buffer containing a primary, secondary or tertiary amine. Said method has been applied, for example, to interleukin-4 and bovine pancreatic trypsin inhibitor (BPTI), which were previously (i) solubilized in the presence of guanidinium hydrochloride as chaotronic agent, and (ii) subjected to sulfitolysis.
1. A method for renaturation of Interleukin-4 or Interleukin-4 derivative comprising the step of adding a refolding buffer comprising Tris-(hydroxymethyl)-aminomethane/H2SO4 in a concentration of 1 to 3 Mol/l or triethanolamine/H2SO4 in a concentration of 0.5 to 2 Mol/l to a solution comprising denatured Interleukin-4 or denatured Interleukin-4 derivative at a protein concentration of 250 to 1000 mg/L.
2. The method of claim 1 , wherein the refolding buffer further comprises contains a solubility enhancer.
3. The method of claim 2 , wherein the solubility enhancer is an ion.
4. The method of claim 2 , wherein the solubility enhancer is chloride.
5. The method of claim 1 , wherein the denatured Interleukin-4 or denatured Interleukin-4 derivative is chemically modified or reduced.