Title:
PRODUCTION OF RECOMBINANT PROTEINS UTILIZING NON-ANTIBIOTIC SELECTION METHODS AND THE INCORPORATION OF NON-NATURAL AMINO ACIDS THEREIN
Document Type and Number:
WIPO Patent Application WO/2010/132341
Kind Code:
A3
Abstract:
Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analogue into the growth media for the host cell such that the analogue is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analogue.
Inventors:
RETALLACK DIANE M (US)
CHEW LAWRENCE C (US)
SQUIRES CHARLES H (US)
CHEW LAWRENCE C (US)
SQUIRES CHARLES H (US)
Application Number:
PCT/US2010/034201
Publication Date:
April 21, 2011
Filing Date:
May 10, 2010
Export Citation:
Assignee:
PFENEX INC (US)
RETALLACK DIANE M (US)
CHEW LAWRENCE C (US)
SQUIRES CHARLES H (US)
RETALLACK DIANE M (US)
CHEW LAWRENCE C (US)
SQUIRES CHARLES H (US)
International Classes:
C12N15/65; C12P21/04
Foreign References:
| US20060183198A1 | 2006-08-17 |
Other References:
SCHNEIDER, J. C. ET AL., BIOTECHNOL. PROG., vol. 21, 14 December 2004 (2004-12-14), pages 343 - 348, XP002421514
PAVELKA, M. S. ET AL., JOURNAL OF BACTERIOLOGY., vol. 181, no. 16, August 1999 (1999-08-01), pages 4780 - 4789, XP002992817
MICHIELSE, C. B. ET AL., MOL. GEN. GENOMICS., vol. 271, 6 April 2004 (2004-04-06), pages 499 - 510, XP008164711
PRONK, J. T. ET AL., APPLIED AND ENVIRONMENTAL MICROBILOGY., vol. 68, no. 5, May 2002 (2002-05-01), pages 2095 - 2100, XP008164715
See also references of EP 2430164A4
PAVELKA, M. S. ET AL., JOURNAL OF BACTERIOLOGY., vol. 181, no. 16, August 1999 (1999-08-01), pages 4780 - 4789, XP002992817
MICHIELSE, C. B. ET AL., MOL. GEN. GENOMICS., vol. 271, 6 April 2004 (2004-04-06), pages 499 - 510, XP008164711
PRONK, J. T. ET AL., APPLIED AND ENVIRONMENTAL MICROBILOGY., vol. 68, no. 5, May 2002 (2002-05-01), pages 2095 - 2100, XP008164715
See also references of EP 2430164A4
Attorney, Agent or Firm:
VAN DYKE, Timothy, H. (390 N. Orange AvenueOrlando, Florida, US)
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