DK2850100T3 - Rekombinant bakterie værtscelle til proteinekspression. - Google Patents
Rekombinant bakterie værtscelle til proteinekspression. Download PDFInfo
- Publication number
- DK2850100T3 DK2850100T3 DK13723086.8T DK13723086T DK2850100T3 DK 2850100 T3 DK2850100 T3 DK 2850100T3 DK 13723086 T DK13723086 T DK 13723086T DK 2850100 T3 DK2850100 T3 DK 2850100T3
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- ser
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/283—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against Fc-receptors, e.g. CD16, CD32, CD64
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Claims (20)
1. Rekombinant gram-negativ bakteriecelle omfattende: a. et mutant spr-gen kodende for et spr-protein, som har en mutation i en eller flere aminosyrer valgt blandt D133, H145, H157, N31, R62, 170, Q73, C94, S95, V98, Q99, R100, L108, Y115, V135, L136, G140, R144 og G147 og b. en ekspressionsvektor omfattende et gen, som udtrykker eller overudtrykker et eller flere proteiner, der er i stand til at lette proteinfoldning valgt blandt FkpA, Skp eller en kombination af FkpA og Skp, hvor cellen har reduceret Tsp proteinaktivitet i forhold til en vildtype-celle, og hvor cellen ikke omfatter et rekombinant polynukleotid kodende for DsbC, og hvor bakteriecellen er isogen til en vildtype-E. coli-celle, bortset fra det muterede spr-gen og den modifikation, der kræves for at reducere Tsp-proteinaktivitet i forhold til en vildtype-celle.
2. Celle ifølge krav 1, hvor det mutante spr-gen koder for et spr-protein med en eller flere mutationer valgt blandt D133A, H145A, H157A, N31Y, R62C, I70T, Q73R, C94A, S95F, V98E, Q99P, R100G, L108S, Y115F , V135D, V135G, L136P, G140C, R144C og G147C.
3. Celle ifølge krav 1, hvor det mutante spr-gen koder for et spr-protein med mutationerne C94 og H145A.
4. Celle ifølge krav 1, hvor det mutante spr-gen koder for et spr-protein med en mutation valgt blandt D133A, H145A og H157A.
5. Celle ifølge krav 1, hvor det mutante spr-gen koder for et spr-protein med en mutation valgt blandt C94A.
6. Celle ifølge ethvert af de foregående krav, hvor det mutante spr-gen er integreret i cellens genom.
7. Celle ifølge ethvert af de foregående krav, hvor et gen kodende for et protein, som er i stand til at lette proteinfoldning, midlertidigt transficeres i cellen.
8. Celle ifølge ethvert af de foregående krav, hvor proteinet, som er i stand til at lette proteinfoldning, er FkpA.
9. Celle ifølge ethvert af de foregående krav, hvor cellen desuden omfatter et eller flere af de følgende muterede gener: et muteret DegP-gen, der koder for et DegP-protein, der har chaperonaktivitet og reduceret proteaseaktivitet; b. et muteret ptr-gen, hvor det muterede ptr-gen koder for et Protease Ill-protein med reduceret proteaseaktivitet eller er et deaktiveret ptr-gen; og c. et muteret OmpT-gen, hvor det muterede OmpT-gen koder for et OmpT-protein med reduceret proteaseaktivitet eller er et deaktiveret OmpT-gen.
10. Celle ifølge ethvert af de foregående krav, hvor cellen omfatter et muteret Tsp-gen, som koder for et Tsp-protein, der har reduceret proteaseaktivitet eller er et deaktiveret Tsp-gen.
11. Celle ifølge ethvert af de foregående krav, hvor cellen har et deaktiveret Tsp-gen, som omfatter en mutation til genstartkodonet og / eller et eller flere stopkodoner placeret nedstrøms for genstartkodonet og opstrøms for genstopkodonet.
12. Celle ifølge krav 11, hvor det deaktiverede Tsp-gen omfatter et restriktionsmarkørsted skabt af en missense-mutation til genstartkodonet og eventuelt en eller flere yderligere punktmutationer.
13. Celle ifølge krav 12, hvor det deaktiverede Tsp-gen omfatter SEQ ID NO: 3.
14. Celle ifølge ethvert af de foregående krav, hvor cellen er E. coli.
15. Celle ifølge krav 14, hvor E. coli er stamme K12 eller W3110.
16. Celle ifølge ethvert af de foregående krav, hvor cellen omfatter en polynu-kleotidsekvens kodende for et protein af interesse valgt blandt et antistof eller et antigenbindende fragment deraf.
17. Celle ifølge krav 16, hvor antistoffet eller det antigenbindende fragment deraf er specifikt for FcRn.
18. Fremgangsmåde til fremstilling af et protein af interesse, der omfatter at dyrke en rekombinant gram-negativ bakteriecelle ifølge ethvert af kravene 1 til 17 i et kulturmedium under betingelser, der er effektive til at udtrykke det rekombinante protein af interesse og at udvinde det rekombinante protein af interesse fra periplasmaet af den rekombinante gram-negative bakteriecelle og / eller dyrkningsmediet.
19. Fremgangsmåde ifølge krav 18, hvor fremgangsmåden yderligere omfatter at udvinde proteinet af interesse fra cellen.
20. Fremgangsmåde ifølge krav 19, hvor proteinet af interesse udvindes fra periplasmaet og / eller supernatanten.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB201208367A GB201208367D0 (en) | 2012-05-14 | 2012-05-14 | Biological product |
| PCT/EP2013/059803 WO2013171156A1 (en) | 2012-05-14 | 2013-05-13 | Recombinant bacterial host cell for protein expression |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2850100T3 true DK2850100T3 (da) | 2019-02-04 |
Family
ID=46458730
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK13723086.8T DK2850100T3 (da) | 2012-05-14 | 2013-05-13 | Rekombinant bakterie værtscelle til proteinekspression. |
Country Status (30)
| Country | Link |
|---|---|
| US (2) | US9951365B2 (da) |
| EP (1) | EP2850100B1 (da) |
| JP (2) | JP6420759B2 (da) |
| KR (1) | KR102036399B1 (da) |
| CN (2) | CN110591991A (da) |
| AU (1) | AU2013261915B2 (da) |
| BR (1) | BR112014027357A2 (da) |
| CA (1) | CA2872328C (da) |
| CL (1) | CL2014003112A1 (da) |
| CO (1) | CO7240426A2 (da) |
| CY (1) | CY1121192T1 (da) |
| DK (1) | DK2850100T3 (da) |
| EA (1) | EA201492100A1 (da) |
| ES (1) | ES2704425T3 (da) |
| GB (1) | GB201208367D0 (da) |
| HR (1) | HRP20190025T1 (da) |
| HU (1) | HUE040624T2 (da) |
| IL (1) | IL235041B (da) |
| LT (1) | LT2850100T (da) |
| ME (1) | ME03309B (da) |
| MX (1) | MX353066B (da) |
| MY (1) | MY166223A (da) |
| PL (1) | PL2850100T3 (da) |
| PT (1) | PT2850100T (da) |
| RS (1) | RS58219B1 (da) |
| SG (1) | SG11201406474RA (da) |
| SI (1) | SI2850100T1 (da) |
| SM (1) | SMT201900011T1 (da) |
| TR (1) | TR201900073T4 (da) |
| WO (1) | WO2013171156A1 (da) |
Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9109216B2 (en) | 2009-09-24 | 2015-08-18 | Ucb Pharma, S.A. | Bacterial host strain |
| 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 |
| GB201012599D0 (en) | 2010-07-27 | 2010-09-08 | Ucb Pharma Sa | Process for purifying proteins |
| HRP20180226T1 (hr) | 2011-07-13 | 2018-03-09 | Ucb Biopharma Sprl | Bakterijski soj domaćina koji eksprimira rekombinantnu dsbc |
| GB201208367D0 (en) * | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
| WO2018119142A1 (en) | 2016-12-21 | 2018-06-28 | Amgen Inc. | Anti-tnf alpha antibody formulations |
| GB201708655D0 (en) | 2017-05-31 | 2017-07-12 | Ucb Biopharma Sprl | Cell culture methods |
| EP3492582A1 (en) | 2017-12-01 | 2019-06-05 | UCB Biopharma SPRL | Cell culture methods |
| EP3774926A1 (en) | 2018-04-05 | 2021-02-17 | Bio-Rad ABD Serotec GmbH | Display systems for proteins of interest |
| US12404512B2 (en) | 2018-05-01 | 2025-09-02 | Ambrx, Inc. | Method for optimizing antibody expression |
| EP3877407B1 (en) * | 2018-11-05 | 2026-03-11 | 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 |
| AU2023281650A1 (en) | 2022-05-30 | 2024-10-17 | Hanall Biopharma Co., Ltd. | Anti-fcrn antibody or antigen binding fragment thereof with improved stability |
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| US5264365A (en) | 1990-11-09 | 1993-11-23 | Board Of Regents, The University Of Texas System | Protease-deficient bacterial strains for production of proteolytically sensitive polypeptides |
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| US6083715A (en) | 1997-06-09 | 2000-07-04 | Board Of Regents, The University Of Texas System | Methods for producing heterologous disulfide bond-containing polypeptides in bacterial cells |
| PT1124941E (pt) | 1998-10-28 | 2004-02-27 | Genentech Inc | Processo para recuperar polipeptidos heterologos a partir de celulas bacterianas |
| DE60031533D1 (de) | 1999-06-29 | 2006-12-07 | Siga Technologies Inc | Test für degp-protease inhibitoren |
| GB0006398D0 (en) | 2000-03-16 | 2000-05-03 | Novartis Ag | Organic compounds |
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| US7041479B2 (en) | 2000-09-06 | 2006-05-09 | The Board Of Trustess Of The Leland Stanford Junior University | Enhanced in vitro synthesis of active proteins containing disulfide bonds |
| JP4056880B2 (ja) | 2000-12-14 | 2008-03-05 | ジェネンテック・インコーポレーテッド | 細菌性宿主株 |
| WO2002061090A2 (en) * | 2000-12-14 | 2002-08-08 | Genentech, Inc. | Prokaryotically produced antibodies and uses thereof |
| TWI334439B (en) | 2001-08-01 | 2010-12-11 | Centocor Inc | Anti-tnf antibodies, compositions, methods and uses |
| CN100513414C (zh) | 2001-08-27 | 2009-07-15 | 杰南技术公司 | 进行抗体表达和装配的系统 |
| GB0124317D0 (en) | 2001-10-10 | 2001-11-28 | Celltech R&D Ltd | Biological products |
| MXPA04004634A (es) | 2001-11-16 | 2004-08-12 | Idec Pharma Corp | Expresion policistronica de anticuerpos. |
| GB0129105D0 (en) | 2001-12-05 | 2002-01-23 | Celltech R&D Ltd | Expression control using variable intergenic sequences |
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| US9109216B2 (en) | 2009-09-24 | 2015-08-18 | Ucb Pharma, S.A. | Bacterial host strain |
| US8470552B2 (en) | 2009-10-12 | 2013-06-25 | Keck Graduate Institute | Strategy to reduce lactic acid production and control PH in animal cell culture |
| EP2496601B1 (en) | 2009-11-05 | 2017-06-07 | F. Hoffmann-La Roche AG | Methods and composition for secretion of heterologous polypeptides |
| GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
| GB201000591D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist 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 |
| HRP20180226T1 (hr) * | 2011-07-13 | 2018-03-09 | Ucb Biopharma Sprl | Bakterijski soj domaćina koji eksprimira rekombinantnu dsbc |
| EP2546267A1 (en) * | 2011-07-13 | 2013-01-16 | UCB Pharma S.A. | Bacterial host strain expressing recombinant DsbC |
| GB201208367D0 (en) * | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
-
2012
- 2012-05-14 GB GB201208367A patent/GB201208367D0/en not_active Ceased
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2013
- 2013-05-13 ME MEP-2019-3A patent/ME03309B/me unknown
- 2013-05-13 ES ES13723086T patent/ES2704425T3/es active Active
- 2013-05-13 KR KR1020147032673A patent/KR102036399B1/ko not_active Expired - Fee Related
- 2013-05-13 PT PT13723086T patent/PT2850100T/pt unknown
- 2013-05-13 CA CA2872328A patent/CA2872328C/en active Active
- 2013-05-13 US US14/400,068 patent/US9951365B2/en active Active
- 2013-05-13 DK DK13723086.8T patent/DK2850100T3/da active
- 2013-05-13 PL PL13723086T patent/PL2850100T3/pl unknown
- 2013-05-13 SG SG11201406474RA patent/SG11201406474RA/en unknown
- 2013-05-13 JP JP2015512012A patent/JP6420759B2/ja not_active Expired - Fee Related
- 2013-05-13 AU AU2013261915A patent/AU2013261915B2/en not_active Ceased
- 2013-05-13 LT LTEP13723086.8T patent/LT2850100T/lt unknown
- 2013-05-13 EA EA201492100A patent/EA201492100A1/ru unknown
- 2013-05-13 MX MX2014012623A patent/MX353066B/es active IP Right Grant
- 2013-05-13 HR HRP20190025TT patent/HRP20190025T1/hr unknown
- 2013-05-13 EP EP13723086.8A patent/EP2850100B1/en active Active
- 2013-05-13 SM SM20190011T patent/SMT201900011T1/it unknown
- 2013-05-13 HU HUE13723086A patent/HUE040624T2/hu unknown
- 2013-05-13 TR TR2019/00073T patent/TR201900073T4/tr unknown
- 2013-05-13 BR BR112014027357A patent/BR112014027357A2/pt active Search and Examination
- 2013-05-13 SI SI201331318T patent/SI2850100T1/sl unknown
- 2013-05-13 WO PCT/EP2013/059803 patent/WO2013171156A1/en not_active Ceased
- 2013-05-13 RS RS20190013A patent/RS58219B1/sr unknown
- 2013-05-13 CN CN201910693072.1A patent/CN110591991A/zh active Pending
- 2013-05-13 CN CN201380025191.0A patent/CN104487454A/zh active Pending
- 2013-05-13 MY MYPI2014703261A patent/MY166223A/en unknown
-
2014
- 2014-10-07 IL IL23504114A patent/IL235041B/en active IP Right Grant
- 2014-11-14 CL CL2014003112A patent/CL2014003112A1/es unknown
- 2014-12-11 CO CO14272157A patent/CO7240426A2/es unknown
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2018
- 2018-04-23 US US15/959,498 patent/US11261470B2/en not_active Expired - Fee Related
- 2018-07-12 JP JP2018132619A patent/JP2018198597A/ja not_active Abandoned
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2019
- 2019-01-10 CY CY20191100022T patent/CY1121192T1/el unknown
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