ME02752B - Bakterijski soj domaćina sa ekspresijom rekombinantne dsbc i imajući smanjenu aktivnost tsp - Google Patents
Bakterijski soj domaćina sa ekspresijom rekombinantne dsbc i imajući smanjenu aktivnost tspInfo
- Publication number
- ME02752B ME02752B MEP-2017-127A MEP12717A ME02752B ME 02752 B ME02752 B ME 02752B ME P12717 A MEP12717 A ME P12717A ME 02752 B ME02752 B ME 02752B
- Authority
- ME
- Montenegro
- Prior art keywords
- protein
- cell
- gene
- seq
- interest
- Prior art date
Links
Classifications
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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 [IG], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/24—Immunoglobulins [IG], 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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- 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
- C12N1/00—Microorganisms; 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
-
- 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
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- 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
-
- 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'
Landscapes
- 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. Rekombinantna gram-negativna bakterijska ćelija je naznačena time, da ćelija: a)obuhvata ekspresioni vektor sadržavajući rekombinantni polinukleotid koji kodira DsbC; b) ima mutirani Tsp gen koji kodira Tsp protein koji ima smanjenu aktivnost proteaze u poređenju sa divljim-tipom ćelije, c) obuhvata mutirani spr gen koji kodira mutirani spr protein sposoban suzbijanju fenotipa ćelije obuhvatajući mutirani Tsp gen, i d) ima genom koji je izogen za divlji- tip E.koli soja W3110 osim za mutirani Tsp gen i mutirani spr gen.
2. Ćelija prema patentnim zahtevu 1, pri čemu mutirani spr gen kodira spr protein, kao što je definisano u SEK ID BR: 21, imajući mutaciju u jednoj ili više aminokiselina izabranih od N31, R62, I70, Q73, C94, S95, V98, Q99, R100, L108, Y115, D133, V135, L136, G140, R144, H145, G147 i H157.
3. Ćelija prema patentnim zahtevu 2, pri čemu mutirani spr gen kodira spr protein, imajući jednu ili više mutacija izabranih od N31Y, R62C, I70T, Q73R, C94A, S95F, V98E, Q99P, R100G, L108S, Y115F, D133A, V135D, V135G, L136P, G140C, R144C, H145A, G147C i H157A.
4. Ćelija prema patentnim zahtevu 3, pri čemu mutirani spr gen kodira spr protein, imajući mutaciju C94A.
5. Ćelija prema patentnim zahtevu 3, pri čemu mutirani spr gen kodira spr protein, imajući mutaciju H145A.
6. Ćelija prema patentnim zahtevu 3, pri čemu mutirani spr gen kodira spr protein, imajući mutacije S95F i Y115F.
7. Ćelija prema bilo kojem od patentnih zahteva 1 do 6, pri čemu ćelija ima nokaut mutirani Tsp gen koji obuhvata mutaciju na genu start kodona i/ili obuhvata jedan ili više stop kodona pozicioniranih nizvodno od start kodona gena i uzvodno od stop kodona gena.
8. Ćelija prema patentnim zahtevu 7, pri čemu nokaut mutirani Tsp gen obuhvata mesto restrikcionog markera nastao pomoću genetske mutacije na start kodonu gena i opciono jednu ili više daljih tačkastih mutacija.
9. Ćelija prema patentnim zahtevu 8, pri čemu nokaut mutirani Tsp gen obuhvata SEK ID BR:3.
10. Ćelija prema bilo kojem od patentnih zahteva 1 do 9, pri čemu ćelija obuhvata polinukleotidnu sekvencu koja kodira protein od interesa.
11. Ćelija prema patentnim zahtevu 10, pri čemu ćelija obuhvata vektor obuhvatajući rekombinantni polinukleotid koji kodira DsbC i polinukleotidnu sekvencu koja kodira protein od interesa.
12. Ćelija prema patentnim zahtevu 11, pri čemu vektor sadrži promoter koji kontroliše ekspresiju rekombinantnog polinukleotida kodirajući DsbC i polinukleotidnu sekvencu koja kodira protein od interesa.
13. Ćelija prema bilo kojem od patentnih zahteva 10 do 12, pri čemu je protein od interesa antitelo ili njegov antigen vezujući fragment.
14. Ćelija prema patentnim zahtevu 13, pri čemu antitelo ili njegov antigen vezujući fragment je Fab, modifikovani Fab ili Fab’.
15. Ćelija prema patentnim zahtevu 13 ili 14, pri čemu je antitelo ili njegov antigen vezujući fragment specifičan za TNF.
16. Ćelija prema patentnim zahtevu 15, pri čemu antitelo ili antigen vezujući fragment obuhvata a) teški lanac pri čemu varijabilni domen obuhvata sekvencu prikazanu u SEK ID BR: 28 za CDRH1, sekvencu prikazanu u SEK ID BR: 29 ili SEK ID BR: 34 za CDRH2 i sekvencu prikazanu u SEK ID BR: 30 za CDRH3 i b) laki lanac pri čemu varijabilni domen sadrži sekvencu prikazanu u SEK ID BR: 31 za CDRL1, sekvencu prikazanu u SEK ID BR: 32 za CDRL2 i sekvencu prikazanu u SEK ID BR: 33 za CDRL3.
17. Ćelija prema patentnim zahtevu 15, pri čemu antitelo ili antigen vezujući fragment je Fab’ i ima sekvencu lakog lanca obuhvatajući ili se sastojeći od SEK ID BR: 13 i sekvence teškog lanca obuhvatajući ili se sastojeći od SEK ID BR: 14.
18. Metoda za proizvodnju proteina od interesa obuhvatajući: a) kultivisanje rekombinantne gram-negativne bakterijske ćelije kao što je definisano u bilo kojem od patentnih zahteva 1 do 17 u medijumu za kulturu pod uslovima efikasnim da izraze rekombinantni protein od interesa i rekombinantni polinukleotid koji kodira DsbC; i b) obnavljanje proteina od interesa iz periplazme rekombinantne gram-negativne bakterijske ćelije i/ili medijuma za kulturu.
19. Metoda prema patentnom zahtevu 18, pri čemu ekspresija polinukleotidne sekvence koja kodira protein od interesa i rekombinantni polinukleotid koji kodira DsbC je indukovan dodavanjem induktora u medijum za kulturu.
20. Metoda prema patentnim zahtevima 18 ili 19, pri čemu metoda dalje obuhvata odvajanje proteina od interesa iz DsbC.
21. Metoda prema bilo kom od patetnih zahteva od 18 do 20, koji nizvodno dalje obuhvata korak obrade PEG-ilacije proteina od interesa.
Applications Claiming Priority (3)
| 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 |
| EP11701045.4A EP2523969B1 (en) | 2010-01-14 | 2011-01-13 | Bacterial host strain expressing recombinant dsbc and having reduced tsp activity |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ME02752B true ME02752B (me) | 2018-01-20 |
Family
ID=42028361
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| MEP-2017-127A ME02752B (me) | 2010-01-14 | 2011-01-13 | Bakterijski soj domaćina sa ekspresijom rekombinantne dsbc i imajući smanjenu aktivnost tsp |
Country Status (23)
| Country | Link |
|---|---|
| US (2) | US8969038B2 (me) |
| EP (1) | EP2523969B1 (me) |
| JP (1) | JP5960608B2 (me) |
| KR (1) | KR101729361B1 (me) |
| CN (2) | CN102712679B (me) |
| AU (2) | AU2011206587A1 (me) |
| CA (1) | CA2786336C (me) |
| CY (1) | CY1118990T1 (me) |
| DK (1) | DK2523969T3 (me) |
| ES (1) | ES2627826T3 (me) |
| GB (1) | GB201000591D0 (me) |
| HK (1) | HK1206785A1 (me) |
| HR (1) | HRP20170852T1 (me) |
| HU (1) | HUE033555T2 (me) |
| LT (1) | LT2523969T (me) |
| ME (1) | ME02752B (me) |
| PL (1) | PL2523969T3 (me) |
| PT (1) | PT2523969T (me) |
| RS (1) | RS56159B1 (me) |
| SG (1) | SG182280A1 (me) |
| SI (1) | SI2523969T1 (me) |
| SM (1) | SMT201700295T1 (me) |
| WO (1) | WO2011086139A1 (me) |
Families Citing this family (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2774692C (en) | 2009-09-24 | 2017-12-19 | Ucb Pharma S.A. | Bacterial strain for recombinant protein expression, having protease deficient degp retaining chaperone activity, and knocked out tsp and ptr genes |
| 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 |
| 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 |
| SMT201800068T1 (it) * | 2011-07-13 | 2018-03-08 | Ucb Biopharma Sprl | Ceppo ospite batterico che esprime dsbc ricombinante |
| 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 |
| US11306328B2 (en) | 2013-07-26 | 2022-04-19 | President And Fellows Of Harvard College | Genome engineering |
| JP6807327B2 (ja) | 2014-12-22 | 2021-01-06 | ユーシービー バイオファルマ エスアールエル | タンパク質製造 |
| RU2018128784A (ru) | 2016-01-27 | 2020-02-27 | МЕДИММЬЮН, ЭлЭлСи | Способы получения антител с заданным профилем гликозилирования |
| EP3824906A1 (en) | 2016-12-21 | 2021-05-26 | Amgen Inc. | Anti-tnf alpha antibody formulations |
| EP3774926A1 (en) | 2018-04-05 | 2021-02-17 | Bio-Rad ABD Serotec GmbH | Display systems for proteins of interest |
| CN113260626B (zh) | 2018-11-05 | 2024-09-13 | 豪夫迈·罗氏有限公司 | 在原核宿主细胞中产生双链蛋白质的方法 |
| CN113874518B (zh) | 2019-03-18 | 2025-04-25 | 生物辐射Abd瑟罗泰克有限公司 | 保护含SpyTag的周质融合蛋白免于蛋白酶Tsp和OmpT降解 |
| ES3026707T3 (en) * | 2019-08-05 | 2025-06-12 | Wacker Chemie Ag | Bacterial strain for releasing a recombinant protein in a fermentation method |
| KR20240021234A (ko) | 2021-06-10 | 2024-02-16 | 펠리칸 테크놀로지 홀딩스, 인크. | 재조합 단백질 발현을 위한 박테리아 숙주 |
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| GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
| GB201000588D0 (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 |
| SMT201800068T1 (it) | 2011-07-13 | 2018-03-08 | Ucb Biopharma Sprl | Ceppo ospite batterico che esprime dsbc ricombinante |
| 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
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2013
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2015
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