Title:
METHODS AND COMPOSITIONS FOR PRODUCTION OF RECOMBINANT PEPTIDES
Document Type and Number:
WIPO Patent Application WO2003102013
Kind Code:
A3
Abstract:
This invention entails a method of solubilizing and recovering, in bioactive and isolated form with retained native state configuration, a target peptide from a host organism in which the heterologous polypeptide is present in insoluble form. Broadly this method comprises (i) disrupting the host cell to produce a lysate (ii) recovering lysate precipitate containing the polypeptide (iii) resuspending the lysate precipitate in a denaturant-free, non-buffered solubilization solution to produce a solubilization preparation that comprises both sodium hydroxide between about 8 and about 10 mM and the target peptide between about 1 and about 4 mg peptide per ml solubilization solution, wherein the resultant solubilization preparation has a pH of between about 9 and about 11.2; (iv) recovering supernatant from the solubilization preparation containing non-denatured target peptide. Optionally, stabilizing compounds and detergents are employed. The invention further comprises isolated insoluble proteins in bioactive form and native state configuration.
Inventors:
GONZALEZ-VILLASENOR LUCIA IREN (US)
Application Number:
PCT/US2002/005256
Publication Date:
March 03, 2005
Filing Date:
February 22, 2002
Export Citation:
Assignee:
GONZALEZ-VILLASENOR LUCIA IREN (US)
International Classes:
C07K1/113; C07K14/47; C12P21/02; (IPC1-7): A23J1/00
Other References:
ESTAPE D. ET AL.: "Optimized procedures for purification and solubilization of basic fibroblast growth factor inclusion bodies", BIOTECHNOLOGY TECH., vol. 10, no. 7, July 1996 (1996-07-01), pages 481 - 484, XP008040156
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