Title:
INDUCIBLE SELF-CLEAVING PROTEASE TAG AND METHOD OF PURIFYING RECOMBINANT PROTEINS USING THE SAME
Document Type and Number:
WIPO Patent Application WO/2011/034622
Kind Code:
A3
Abstract:
A method of purifying a protein is disclosed which entails: a) fusing a site-specific affinity-tagged cysteine protease domain to a target protein to form a tagged fusion protein; b) activating the site-specific cysteine protease domain of the tagged fusion protein by subjecting the site-specific affinity-tagged cysteine protease domain to an inducer, which induces autoprocessing at a cleavage site; thereby releasing untagged target protein; and c) isolating the untagged target protein.
Inventors:
BOGYO MATTHEW (US)
GARCIA K CHRISTOPHER (US)
SHEN AIMEE (US)
LUPARDUS PATRICK J (US)
GARCIA K CHRISTOPHER (US)
SHEN AIMEE (US)
LUPARDUS PATRICK J (US)
Application Number:
PCT/US2010/002571
Publication Date:
August 09, 2012
Filing Date:
September 21, 2010
Export Citation:
Assignee:
UNIV LELAND STANFORD JUNIOR (US)
BOGYO MATTHEW (US)
GARCIA K CHRISTOPHER (US)
SHEN AIMEE (US)
LUPARDUS PATRICK J (US)
BOGYO MATTHEW (US)
GARCIA K CHRISTOPHER (US)
SHEN AIMEE (US)
LUPARDUS PATRICK J (US)
International Classes:
C07K1/14
Foreign References:
| US20060183121A1 | 2006-08-17 | |||
| US20100137563A1 | 2010-06-03 |
Other References:
PROCHAZKOVA ET AL.: "Structure-function analysis of inositol hexakisphosphate-induced autoprocessing of the Vibrio cholerae multifunctional autoprocessing RTX toxin.", J BIOL CHEM, vol. 283, no. 35, 29 August 2008 (2008-08-29), pages 23656 - 23664, XP055047425, DOI: doi:10.1074/jbc.M803334200
SATCHELL.: "MARTX, multifunctional autoprocessing repeats-in-toxin toxins.", INFECT IMMUN, vol. 75, no. 11, November 2007 (2007-11-01), pages 5079 - 5084
HALL.: "Applying Fusion Protein Technology to E. coli", BIOPHARM INTERNAT, vol. 20, 2007, pages 6 - 14, Retrieved from the Internet
SHEN ET AL.: "Simplified, enhanced protein purification using an inducible, autoprocessing enzyme tag.", PLOS ONE, vol. 4, no. 12, 2 December 2009 (2009-12-02), pages E8119 1 - 11
KEMP ET AL.: "Expression of Plasmodium falciparum blood-stage antigens in Escherichia coli: Detection with antibodies from immune humans.", PROC NAT ACAD SCI, vol. 80, June 1983 (1983-06-01), pages 3787 - 3791
FU ET AL.: "Cloning, expression, purification, and characterization of rat MMP-12.", PROTEIN EXPR PURIF, vol. 21, no. 2, March 2001 (2001-03-01), pages 268 - 274, XP002237748, DOI: doi:10.1006/prep.2000.1376
SATCHELL.: "MARTX, multifunctional autoprocessing repeats-in-toxin toxins.", INFECT IMMUN, vol. 75, no. 11, November 2007 (2007-11-01), pages 5079 - 5084
HALL.: "Applying Fusion Protein Technology to E. coli", BIOPHARM INTERNAT, vol. 20, 2007, pages 6 - 14, Retrieved from the Internet
SHEN ET AL.: "Simplified, enhanced protein purification using an inducible, autoprocessing enzyme tag.", PLOS ONE, vol. 4, no. 12, 2 December 2009 (2009-12-02), pages E8119 1 - 11
KEMP ET AL.: "Expression of Plasmodium falciparum blood-stage antigens in Escherichia coli: Detection with antibodies from immune humans.", PROC NAT ACAD SCI, vol. 80, June 1983 (1983-06-01), pages 3787 - 3791
FU ET AL.: "Cloning, expression, purification, and characterization of rat MMP-12.", PROTEIN EXPR PURIF, vol. 21, no. 2, March 2001 (2001-03-01), pages 268 - 274, XP002237748, DOI: doi:10.1006/prep.2000.1376
Attorney, Agent or Firm:
BEAUMONT, William, E. (Pllc333 N. Fairfax St., Suite 30, Alexandria VA, US)
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