High pressure refolding of protein aggregates and inclusion bodies

US6489450B2 · kind B2 · utility

Assignee

• The Regents of the University of Technology Office of Technology Transfer · US

Inventors

• Theodore W. Randolph · Niwot, US
• John Carpenter · Littleton, US
• Richard St. John · San Francisco, US

Key dates

• Filing date: Jul 9, 1999
• Grant date: Dec 3, 2002
• Expiry date: Jul 9, 2019

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N9/2462
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present disclosure provides an effective method for the refolding of denatured proteins in solution so that properly folded, biologically active protein in solution is recovered in high yield. The refolding takes place at pressures between about 0.25 kbar to about 3.5 kbar, advantageously at about 1.5 kbar to about 3 kbar. Typically a chaotropic agent is present at a concentration which is not effective for denaturing protein at atmospheric pressure, and optionally, oxidation-reduction reagents can be incorporated in the refolding solution so that native intramolecular disulfide bonds can be formed where that is desired. The method is applicable to substantially all proteins, especially after solubilization and/or denaturation of insoluble protein aggregates, inclusion bodies, or abnormal oligomeric (soluble) aggregates.