Back to results
US 11,697,803 B2Granted
Method of protein purification from E.coli
AssigneeAnsun Biopharma, Inc.
InventorStephen Hawley
Life of this patent
Estimating the term…
Abstract
The present disclosure provides methods for releasing intracellular proteins. The method allows isolation of the protein of interest from the cell without the requirement for mechanical disruption of the cells, without the need for isolation of the cells from the culture media, and without the need for removal of the cells from the culture media.
Claims18
A composition comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1 or 2 made by a method comprising:
- (a) providing a liquid culture of a cell expressing the polypeptide;
- (b1) contacting the culture with an inorganic salt;
- holding the culture containing the added inorganic salt for at least 10 minutes;
- contacting the culture containing the added inorganic salt with a chelating agent;
- or
- (b2) contacting the culture with an inorganic salt and a chelating agent;
- (c) holding the culture containing the added inorganic salt and added chelating agent for at least 10 minutes;
- (d) contacting the culture containing the added inorganic salt and the added chelating agent with a detergent;
- (e) contacting the culture containing the added inorganic salt, the added chelating agent and the added detergent with a precipitating agent;
- (f) holding the culture containing the added inorganic salt, the added chelating agent and the added detergent for at least 1 hour;
- and
- (g) subjecting the culture comprising the added inorganic salt, the added chelating agent, the added detergent and the added precipitating agent to a method to remove a substantial portion of the cellular debris, thereby providing a supernatant composition comprising the polypeptide;
- wherein the method does not comprise:
- (i) mechanical disruption of the cells, (ii) removing substantially all of the culture media, or (iii) addition of an enzyme that degrades cell wall material.
12 more elements
Dependencies are inferred from the claim text.