| US 7,538,198 B2 | ||
| Methods for protein refolding | ||
| Theodore W. Randolph, Niwot, Colo. (US); John F. Carpenter, Littleton, Colo. (US); Richard J. St. John, San Francisco, Calif. (US); and Jonathan N. Webb, Zionsville, Ind. (US) | ||
| Assigned to Barofold, Inc., Boulder, Colo. (US) | ||
| Filed on Apr. 29, 2003, as Appl. No. 10/425,371. | ||
| Application 10/425371 is a continuation of application No. PCT/US01/45728, filed on Oct. 31, 2001. | ||
| Claims priority of provisional application 60/244808, filed on Oct. 31, 2000. | ||
| Prior Publication US 2004/0038333 A1, Feb. 26, 2004 | ||
| This patent is subject to a terminal disclaimer. | ||
| Int. Cl. C07K 1/02 (2006.01); C12N 9/00 (2006.01) | ||
| U.S. Cl. 530—427 [435/183] | 42 Claims |
| 1. A method of producing biologically active disaggregated protein from protein aggregates comprising:
(i) providing a protein aggregate;
(ii) mixing said protein aggregate with a reducing agent in an amount sufficient to reduce disulfide bonds therein;
(iii) subjecting the mixture of step (ii) to increased pressure of about 0.25 kbar to about 12 kbar, whereby said protein
aggregate dissolves and/or disaggregates;
(iv) dialyzing the mixture under pressure, whereby said reducing agent is removed and disulfide bonds reform; and
(v) removing the dissolved and/or disaggregated protein from increased pressure, whereby said protein refolds such that biological
activity is retained.
|