Process for protein extraction
AssigneeGenentech, Inc.
InventorRichard L. Gehant
Life of this patent
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Abstract
The invention includes a process for extracting a target protein from Escherichia coli cells that includes lowering the pH of a whole E. coli cell solution to form an acidic solution, disrupting the cells to release the protein into the acidic solution, and separating the cellular debris from the released protein to obtain a protein product enriched in the heterologous target protein. The invention also includes addition of a solubility enhancer.
Claims48
A recombinant protein product produced by a method for extracting a heterologous target protein from Escherichia coli (E. coli) cells, comprising the following steps:
- a) lowering the pH of a solution containing whole E. coli cells expressing said heterologous target protein to form an acidic cell solution;
- b) adding at least one solubility enhancer to the solution containing the E. coli cells, wherein said at least one solubility enhancer is added to the solution prior to or contemporaneously with said pH lowering in step a);
- c) disrupting the cells to release the heterologous target protein into the acidic solution;
- and
- d) separating cellular debris from the released heterologous target protein to obtain a protein preparation enriched in the heterologous target protein.
A recombinant protein product produced by a method for extracting a heterologous target protein from a solution of disrupted E. coli cells expressing the heterologous target protein, comprising the steps of:
- a) lowering the pH of the solution to be acidic;
- b) adding at least one solubility enhancer to the solution, wherein the solubility enhancer comprises a divalent cation or polyethyleneimine (PEI);
- and
- c) separating cellular debris from the heterologous target protein.
Dependencies are inferred from the claim text.