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1. (WO2007002532) CELL LINES FOR EXPRESSING ENZYME USEFUL IN THE PREPARATION OF AMIDATED PRODUCTS
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/2007/002532 International Application No.: PCT/US2006/024687
Publication Date: 04.01.2007 International Filing Date: 23.06.2006
Chapter 2 Demand Filed: 22.01.2007
IPC:
C12P 21/04 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
21
Preparation of peptides or proteins
02
having a known sequence of two or more amino acids, e.g. glutathione
04
Cyclic or bridged peptides or polypeptides, e.g. bacitracin
Applicants:
UNIGENE LABORATORIES, INC. [US/US]; 110 Little Falls Road Fairfield, NJ 07004-2193, US (AllExceptUS)
MILLER, Duncan, A. [US/US]; US (UsOnly)
MEHTA, Nozer, M. [US/US]; US (UsOnly)
CONSALVO, Angelo, P. [US/US]; US (UsOnly)
Inventors:
MILLER, Duncan, A.; US
MEHTA, Nozer, M.; US
CONSALVO, Angelo, P.; US
Agent:
GRAY, William, O., III ; Ostrolenk, Faber, Gerb & Soffen, LLP 1180 Avenue of the Americas New York, NY 10036, US
Priority Data:
60/693,61224.06.2005US
Title (EN) CELL LINES FOR EXPRESSING ENZYME USEFUL IN THE PREPARATION OF AMIDATED PRODUCTS
(FR) LIGNEES CELLULAIRES PERMETTANT D'EXPRIMER UNE ENZYME UTILE POUR LA PREPARATION DE PRODUITS AMIDES
Abstract:
(EN) Cell lines are provided for expressing peptidylglycine alpha-amidating monooxygenase (PAM), or one of its two catalytic domains. High levels of enzyme expression are achieved while utilizing a non-animal source, low protein tissue culture medium. A robust two-step downstream purification results in high enzyme purity. Resulting PAM, or its PHM catalytic domain, is used to catalyze the enzymatic conversion of X-Gly to X-alpha-hydroxy-Gly or X-NH2 (X being a peptide or any chemical compound having a carbonyl group to which a glycine group can be covalently attached). Methods of preparing preferred cell lines are also set forth.
(FR) L'invention porte sur des lignées cellulaires qui permettent d'exprimer la peptidylglycine alpha-amidating monooxygénase (PAM) ou l'un de ses deux domaines catalytiques. On obtient des niveaux élevés d'expression enzymatique tout en utilisant un milieu de culture tissulaire à faible teneur en protéines de source non animale. Une purification aval robuste en deux étapes permet d'arriver à une haute pureté enzymatique. On utilise la PAM obtenue, ou son domaine catalytique PHM, pour catalyser la conversion enzymatique de X-Gly en X-alpha-hydroxy-Gly ou en X-NH2 (X étant un peptide ou un quelconque composé chimique renfermant un groupe carbonyle auquel un groupe glycine peut être attaché par covalence). L'invention concerne également des procédés de préparation de lignées cellulaires préférées.
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Designated States: AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BW, BY, BZ, CA, CH, CN, CO, CR, CU, CZ, DE, DK, DM, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LV, LY, MA, MD, MG, MK, MN, MW, MX, MZ, NA, NG, NI, NO, NZ, OM, PG, PH, PL, PT, RO, RS, RU, SC, SD, SE, SG, SK, SL, SM, SY, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Organization (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM)
European Patent Office (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HU, IE, IS, IT, LT, LU, LV, MC, NL, PL, PT, RO, SE, SI, SK, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG)
Publication Language: English (EN)
Filing Language: English (EN)
Also published as:
EP1907561JP2008543344JP2011139714JP2012016354CN101287840CA2613471
ES2437068AU2006261909DK1907561