DK2523969T3 - Bakterieværtsstamme, som udtrykker rekombinant DsbC og har nedsat Tsp-aktivitet - Google Patents
Bakterieværtsstamme, som udtrykker rekombinant DsbC og har nedsat Tsp-aktivitet Download PDFInfo
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- DK2523969T3 DK2523969T3 DK11701045.4T DK11701045T DK2523969T3 DK 2523969 T3 DK2523969 T3 DK 2523969T3 DK 11701045 T DK11701045 T DK 11701045T DK 2523969 T3 DK2523969 T3 DK 2523969T3
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K14/245—Escherichia (G)
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/241—Tumor Necrosis Factors
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/90—Isomerases (5.)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y503/00—Intramolecular oxidoreductases (5.3)
- C12Y503/04—Intramolecular oxidoreductases (5.3) transposing S-S bonds (5.3.4)
- C12Y503/04001—Protein disulfide-isomerase (5.3.4.1), i.e. disufide bond-forming enzyme
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/55—Fab or Fab'
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Claims (21)
1. Rekombinant gramnegativ bakteriecelle, kendetegnet ved, at cellen: a) omfatter en ekspressionsvektor, der omfatter et rekombinant polynukleotid kodende for DsbC; b) har et muteret Tsp-gen kodende for et Tsp-protein med nedsat proteaseakti-vitet i sammenligning med en vildtypecelle, c) omfatter et muteret spr-gen kodende for et mutant spr-protein med evne til at undertrykke fænotypen af en celle omfattende et muteret Tsp-gen, og d) har et genom, som er isogent med vildtypestammen W3110 af E. coli, bortset fra det muterede Tsp-gen og det muterede spr-gen.
2. Celle ifølge krav 1, hvor det muterede spr-gen koder for et spr-protein som defineret i SEQ ID NO: 21 med en mutation i en eller flere aminosyrer udvalgt blandt N31, R62, 170, Q73, C94, S95, V98, Q99, R100, L108, Y115, D133, V135, L136, G140, R144, H145, G147 og H157.
3. Celle ifølge krav 2, hvor det mutante spr-gen koder for et spr-protein med en eller flere mutationer udvalgt blandt N31Y, R62C, I70T, Q73R, C94A, S95F, V98E, Q99P, R100G, L108S, Y115F, D133A, V135D, V135G, L136P, G140C, R144C, H145A, G147C og H157A.
4. Celle ifølge krav 3, hvor det mutante spr-gen koder for et spr-protein med mutationen C94A.
5. Celle ifølge krav 3, hvor det mutante spr-gen koder for et spr-protein med mutationen H145A.
6. Celle ifølge krav 3, hvor det mutante spr-gen koder for et spr-protein med mutationerne S95F og Y115F.
7. Celle ifølge et hvilket som helst af kravene 1 til 6, hvor cellen har et knockout-muteret Tsp-gen, som omfatter en mutation i genstartkodonen og/eller omfatter en eller flere stopkodoner placeret nedstrøms for genstartkodonen og opstrøms for genstopkodonen.
8. Celle ifølge krav 7, hvor det knockout-muterede Tsp-gen omfatter et restriktionsmarkørsted dannet af en missense-mutation i genstartkodonen og eventuelt en eller flere punktmutationer.
9. Celle ifølge krav 8, hvor det knockout-muterede Tsp-gen omfatter SEQ ID NO:3.
10. Celle ifølge et hvilket som helst af kravene 1 til 9, hvor cellen omfatter en polynu-kleotidsekvens kodende for et protein af interesse.
11. Celle ifølge krav 10, hvor cellen omfatter en vektor omfattende det rekombinante polynukleotid kodende for DsbC og polynukleotidsekvensen kodende for et protein af interesse.
12. Celle ifølge krav 11, hvor vektoren omfatter en promotor, der styrer udtrykkeisen af det rekombinante polynukleotid kodende for DsbC og polynukleotidsekvensen kodende for et protein af interesse.
13. Celle ifølge et hvilket som helst af kravene 10 til 12, hvor proteinet af interesse er et antistof eller et antigenbindende fragment deraf.
14. Celle ifølge krav 13, hvor antistoffet eller det antigenbindende fragment deraf er et Fab, modificeret Fab eller Fab'.
15. Celle ifølge krav 13 eller 14, hvor antistoffet eller det antigenbindende fragment deraf er specifikt for TNF.
16. Celle ifølge krav 15, hvor antistoffet eller et antigenbindende fragment omfatter a) en tung kæde, hvori det variable domæne omfatter sekvensen vist i SEQ ID NO:28 for CDRH1, sekvensen vist i SEQ ID NO:29 eller SEQ ID NO:34 for CDRH2 og sekvensen vist i SEQ ID NO:30 for CDRFI3 og b) en let kæde, hvor det variable domæne omfatter sekvensen vist i SEQ ID NO:31 for CDRL1, sekvensen vist i SEQ ID NO:32 for CDRL2 og sekvensen vist i SEQ ID NO:33 for CDRL3.
17. Celle ifølge krav 15, hvor antistoffet eller et antigenbindende fragment er et Fab' og har en letkædesekvens omfattende eller bestående af SEQ ID NO: 13 og en tungkædese-kvens omfattende eller bestående af SEQ ID NO: 14.
18. Fremgangsmåde til fremstilling af et protein af interesse, hvilken fremgangsmåde omfatter: a) at dyrke en rekombinant gramnegativ bakteriecelle som defineret i et hvilket som helst af kravene 1 til 17 i et kulturmedium under betingelser, der er virksomme til udtryk-kelse af det rekombinante protein af interesse og det rekombinante polynukleotid kodende for DsbC; og b) at indvinde proteinet af interesse fra periplasmaet af den rekombinante gramnegative bakteriecelle og/eller kulturmediet.
19. Fremgangsmåde ifølge krav 18, hvor udtrykkeisen af polynukleotidsekvensen kodende for et protein af interesse og det rekombinante polynukleotid kodende for DsbC fremkaldes af tilsættelse af et induktionsmiddel til kulturmediet.
20. Fremgangsmåde ifølge krav 18 eller 19, hvor fremgangsmåden yderligere omfatter at separere proteinet af interesse fra DsbC.
21. Fremgangsmåde ifølge et hvilket som helst af kravene 18 til 20, hvilken fremgangsmåde yderligere omfatter PEGylering af proteinet af interesse som nedstrøms forarbejdningstrin.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GBGB1000591.6A GB201000591D0 (en) | 2010-01-14 | 2010-01-14 | Bacterial hoist strain |
PCT/EP2011/050416 WO2011086139A1 (en) | 2010-01-14 | 2011-01-13 | Bacterial host strain expressing recombinant dsbc and having reduced tsp activity |
Publications (1)
Publication Number | Publication Date |
---|---|
DK2523969T3 true DK2523969T3 (da) | 2017-06-12 |
Family
ID=42028361
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DK11701045.4T DK2523969T3 (da) | 2010-01-14 | 2011-01-13 | Bakterieværtsstamme, som udtrykker rekombinant DsbC og har nedsat Tsp-aktivitet |
Country Status (23)
Country | Link |
---|---|
US (2) | US8969038B2 (da) |
EP (1) | EP2523969B1 (da) |
JP (1) | JP5960608B2 (da) |
KR (1) | KR101729361B1 (da) |
CN (2) | CN102712679B (da) |
AU (2) | AU2011206587A1 (da) |
BR (1) | BR112012016808A2 (da) |
CA (1) | CA2786336C (da) |
CY (1) | CY1118990T1 (da) |
DK (1) | DK2523969T3 (da) |
ES (1) | ES2627826T3 (da) |
GB (1) | GB201000591D0 (da) |
HK (2) | HK1176943A1 (da) |
HR (1) | HRP20170852T1 (da) |
HU (1) | HUE033555T2 (da) |
LT (1) | LT2523969T (da) |
ME (1) | ME02752B (da) |
PL (1) | PL2523969T3 (da) |
PT (1) | PT2523969T (da) |
RS (1) | RS56159B1 (da) |
SG (1) | SG182280A1 (da) |
SI (1) | SI2523969T1 (da) |
WO (1) | WO2011086139A1 (da) |
Families Citing this family (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
HUE040306T2 (hu) | 2009-09-24 | 2019-03-28 | Ucb Biopharma Sprl | Baktériumtörzs rekombináns fehérje expresszálására, amely tartalmaz proteázhiányos, de chaperonaktivitását megtartott DEGP-t és génkiütött TSP és PTR gént |
GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
GB201000587D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201000591D0 (en) * | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201001791D0 (en) | 2010-02-03 | 2010-03-24 | Ucb Pharma Sa | Process for obtaining antibodies |
GB201012599D0 (en) * | 2010-07-27 | 2010-09-08 | Ucb Pharma Sa | Process for purifying proteins |
EP2546267A1 (en) * | 2011-07-13 | 2013-01-16 | UCB Pharma S.A. | Bacterial host strain expressing recombinant DsbC |
HUE035674T2 (en) | 2011-07-13 | 2018-05-28 | Ucb Biopharma Sprl | Bacterial host strains expressing recombinant DSBC |
GB201208367D0 (en) | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
US10563225B2 (en) | 2013-07-26 | 2020-02-18 | President And Fellows Of Harvard College | Genome engineering |
EP4253548A3 (en) | 2014-12-22 | 2024-01-03 | UCB Biopharma SRL | Protein manufacture |
AU2017212484C1 (en) | 2016-01-27 | 2020-11-05 | Medimmune, Llc | Methods for preparing antibodies with a defined glycosylation pattern |
EP3877407A1 (en) * | 2018-11-05 | 2021-09-15 | F. Hoffmann-La Roche AG | Methods of producing two chain proteins in prokaryotic host cells |
EP3942079A1 (en) | 2019-03-18 | 2022-01-26 | Bio-Rad ABD Serotec GmbH | Protection of spytag-containing periplasmic fusion proteins from protease tsp and ompt degradation |
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GB201000591D0 (en) * | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
GB201000587D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201000588D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
GB201001791D0 (en) | 2010-02-03 | 2010-03-24 | Ucb Pharma Sa | Process for obtaining antibodies |
GB201012599D0 (en) | 2010-07-27 | 2010-09-08 | Ucb Pharma Sa | Process for purifying proteins |
EP2546267A1 (en) | 2011-07-13 | 2013-01-16 | UCB Pharma S.A. | Bacterial host strain expressing recombinant DsbC |
HUE035674T2 (en) | 2011-07-13 | 2018-05-28 | Ucb Biopharma Sprl | Bacterial host strains expressing recombinant DSBC |
GB201208367D0 (en) | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
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2010
- 2010-01-14 GB GBGB1000591.6A patent/GB201000591D0/en not_active Ceased
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2011
- 2011-01-13 PT PT117010454T patent/PT2523969T/pt unknown
- 2011-01-13 EP EP11701045.4A patent/EP2523969B1/en active Active
- 2011-01-13 WO PCT/EP2011/050416 patent/WO2011086139A1/en active Application Filing
- 2011-01-13 CN CN201180005978.1A patent/CN102712679B/zh active Active
- 2011-01-13 CN CN201410591520.4A patent/CN104328079A/zh active Pending
- 2011-01-13 AU AU2011206587A patent/AU2011206587A1/en not_active Abandoned
- 2011-01-13 RS RS20170626A patent/RS56159B1/sr unknown
- 2011-01-13 DK DK11701045.4T patent/DK2523969T3/da active
- 2011-01-13 SG SG2012047676A patent/SG182280A1/en unknown
- 2011-01-13 BR BR112012016808A patent/BR112012016808A2/pt not_active Application Discontinuation
- 2011-01-13 PL PL11701045T patent/PL2523969T3/pl unknown
- 2011-01-13 JP JP2012548441A patent/JP5960608B2/ja active Active
- 2011-01-13 ES ES11701045.4T patent/ES2627826T3/es active Active
- 2011-01-13 KR KR1020127021021A patent/KR101729361B1/ko active IP Right Grant
- 2011-01-13 US US13/522,083 patent/US8969038B2/en active Active
- 2011-01-13 SI SI201131205A patent/SI2523969T1/sl unknown
- 2011-01-13 ME MEP-2017-127A patent/ME02752B/me unknown
- 2011-01-13 LT LTEP11701045.4T patent/LT2523969T/lt unknown
- 2011-01-13 CA CA2786336A patent/CA2786336C/en active Active
- 2011-01-13 HU HUE11701045A patent/HUE033555T2/hu unknown
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2013
- 2013-03-25 HK HK13103681.3A patent/HK1176943A1/xx unknown
- 2013-03-25 HK HK15107468.1A patent/HK1206785A1/xx unknown
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2015
- 2015-02-27 US US14/633,294 patent/US9493559B2/en active Active
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2017
- 2017-04-12 AU AU2017202434A patent/AU2017202434B2/en active Active
- 2017-06-05 HR HRP20170852TT patent/HRP20170852T1/hr unknown
- 2017-06-22 CY CY20171100667T patent/CY1118990T1/el unknown
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