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US 6,605,706 B1Granted
Method for producing a correctly folded, biological active recombinant protein
AssigneePharmacia Corporation
InventorsJan-Gunnar Gustafsson, Johan Öhman
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Abstract
Methods for producing correctly folded biological active recombinant protein or polypeptide comprise the steps of expressing the protein or polypeptide in prokaryotic cells, harvesting the cells, directly solubilizing the cells in a buffer at pH of about 8 to 11 with the chaotropic agent and a reducing agent, and diluting the solution with water and a diluent. The method is conducted in the absence of mechanical disruption of the cells and isolation and washing of refractile bodies.
Claims19
A method for producing a correctly folded, biologically active recombinant protein or polypeptide, comprising the steps of
- a) expressing the protein or polypeptide in prokaryotic cells,
- b) after step a), harvesting the cells by a process comprising centrifugation or cross flow filtration,
- c) after step b), directly solubilizing the cells in a buffer at pH of about 8 to 11 with a chaotropic agent and a reducing agent to form a solution, and
- d) after step c), diluting the solution with water and a diluent, thereby obtaining the correctly folded, biologically active recombinant protein or polypeptide, the method being conducted in the absence of mechanical disruption of the cells and in the absence of isolation and washing of refractile bodies from within the cells.
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Dependencies are inferred from the claim text.