Title:
ENZYME AND PREPARATION METHOD
Document Type and Number:
WIPO Patent Application WO2005087917
Kind Code:
A3
Abstract:
A method for preparing a soluble protein comprising a modified form of urokinase-type plasminogen activator (uPA) or an active fragment thereof, or a variant of either of these which has uPA activity, which method comprises contacting said protein with a buffer at a pH of from 8.5-10.5, said buffer comprising a reducing agent and an oxidising agent which forms a redox pair, wherein the reducing agent is present in excess compared to the oxidising agent, and wherein the reducing agent is present in a concentration of at least 5 mM. Material obtainable in this way forms a further aspect of the invention. It has been refolded in a "native-like" form and is useful in studies such as NMR analysis to detect ligands.
Inventors:
MCALISTER MARK SAMUEL BEATON (GB)
PINEDA-LUCENA ANTONIO (GB)
PINEDA-LUCENA ANTONIO (GB)
Application Number:
PCT/GB2005/000873
Publication Date:
October 27, 2005
Filing Date:
March 07, 2005
Export Citation:
Assignee:
ASTRAZENECA AB (SE)
ASTRAZENECA UK LTD (GB)
MCALISTER MARK SAMUEL BEATON (GB)
PINEDA-LUCENA ANTONIO (GB)
ASTRAZENECA UK LTD (GB)
MCALISTER MARK SAMUEL BEATON (GB)
PINEDA-LUCENA ANTONIO (GB)
International Classes:
C12N9/72; (IPC1-7): C12N9/72; C12N15/58; C12N15/62; C12Q1/37
Domestic Patent References:
| WO2001055174A2 | 2001-08-02 | |||
| WO2004094344A2 | 2004-11-04 |
Foreign References:
| EP0866967A1 | 1998-09-30 |
Other References:
ZESLAWSKA E ET AL: "Crystals of the urokinase type plasminogen activator variant betac-uPA in complex with small molecule inhibitors open the way towards structure-based drug design", JOURNAL OF MOLECULAR BIOLOGY, LONDON, GB, vol. 301, no. 2, 11 August 2000 (2000-08-11), pages 465 - 475, XP004480446, ISSN: 0022-2836
TANG W ET AL: "An Efficient System for Production of Recombinant Urokinase-Type Plasminogen Activator", PROTEIN EXPRESSION AND PURIFICATION, ACADEMIC PRESS, SAN DIEGO, CA, US, vol. 11, no. 3, December 1997 (1997-12-01), pages 279 - 283, XP004459933, ISSN: 1046-5928
HANSEN A P ET AL: "Solution structure of the amino-terminal fragment of urokinase-type plasminogen activator.", BIOCHEMISTRY. 26 APR 1994, vol. 33, no. 16, 26 April 1994 (1994-04-26), pages 4847 - 4864, XP002342850, ISSN: 0006-2960
HAJDUK P J ET AL: "Identification of novel inhibitors of urokinase via NMR-based screening.", JOURNAL OF MEDICINAL CHEMISTRY. 19 OCT 2000, vol. 43, no. 21, 19 October 2000 (2000-10-19), pages 3862 - 3866, XP002341167, ISSN: 0022-2623
TANG W ET AL: "An Efficient System for Production of Recombinant Urokinase-Type Plasminogen Activator", PROTEIN EXPRESSION AND PURIFICATION, ACADEMIC PRESS, SAN DIEGO, CA, US, vol. 11, no. 3, December 1997 (1997-12-01), pages 279 - 283, XP004459933, ISSN: 1046-5928
HANSEN A P ET AL: "Solution structure of the amino-terminal fragment of urokinase-type plasminogen activator.", BIOCHEMISTRY. 26 APR 1994, vol. 33, no. 16, 26 April 1994 (1994-04-26), pages 4847 - 4864, XP002342850, ISSN: 0006-2960
HAJDUK P J ET AL: "Identification of novel inhibitors of urokinase via NMR-based screening.", JOURNAL OF MEDICINAL CHEMISTRY. 19 OCT 2000, vol. 43, no. 21, 19 October 2000 (2000-10-19), pages 3862 - 3866, XP002341167, ISSN: 0022-2623
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