| US 7,544,493 B2 | ||
| Method for the purification of an N-terminal fragment of hepatocyte growth factor | ||
| Friederike Hesse, Munich (Germany); Martin Lanzendoerfer, Tutzing (Germany); Apollon Papadimitriou, Bichl (Germany); and Jan Stracke, Penzberg (Germany) | ||
| Assigned to Hoffmann-La Roche Inc., Nutley, N.J. (US) | ||
| Appl. No. 10/591,040 PCT Filed Mar. 02, 2005, PCT No. PCT/EP2005/002177 § 371(c)(1), (2), (4) Date Aug. 29, 2006, PCT Pub. No. WO2005/095449, PCT Pub. Date Oct. 13, 2005. |
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| Claims priority of application No. 04004950 (EP), filed on Mar. 03, 2004. | ||
| Prior Publication US 2007/0287827 A1, Dec. 13, 2007 | ||
| Int. Cl. C12P 21/04 (2006.01); C12N 1/00 (2006.01); C12N 1/06 (2006.01); C12N 5/00 (2006.01); C12N 15/00 (2006.01); C07K 14/475 (2006.01) | ||
| U.S. Cl. 435—71.1 [435/71.2; 435/320.1; 435/325; 435/243; 435/259; 530/412] | 8 Claims |
| 1. A method for the production of a polypeptide having the amino acid sequence of SEQ ID NO: 2, comprising:
(a) expressing a nucleic acid encoding said polypeptide in a microbial host cell,
(b) isolating inclusion bodies containing said polypeptide in denatured form,
(c) solubilizing the inclusion bodies at a pH of 7-9 in a phosphate buffered solution comprising a denaturing agent, and
(d) renaturing the denatured polypeptide at a pH of 7-9 in a phosphate buffered solution comprising reduced glutathione (GSH)
and oxidized glutathione (GSSG) and a denaturing agent in a non-denaturing concentration.
|