HRP20180226T1 - Bakterijski soj domaćina koji eksprimira rekombinantnu dsbc - Google Patents
Bakterijski soj domaćina koji eksprimira rekombinantnu dsbc Download PDFInfo
- Publication number
- HRP20180226T1 HRP20180226T1 HRP20180226TT HRP20180226T HRP20180226T1 HR P20180226 T1 HRP20180226 T1 HR P20180226T1 HR P20180226T T HRP20180226T T HR P20180226TT HR P20180226 T HRP20180226 T HR P20180226T HR P20180226 T1 HRP20180226 T1 HR P20180226T1
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- Prior art keywords
- seq
- gram
- bacterial cell
- negative bacterial
- antibody
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- 230000001580 bacterial effect Effects 0.000 title claims 17
- 210000004027 cell Anatomy 0.000 claims 16
- 102000040430 polynucleotide Human genes 0.000 claims 12
- 108091033319 polynucleotide Proteins 0.000 claims 12
- 239000002157 polynucleotide Substances 0.000 claims 12
- 239000000427 antigen Substances 0.000 claims 9
- 102000036639 antigens Human genes 0.000 claims 9
- 108091007433 antigens Proteins 0.000 claims 9
- 239000012634 fragment Substances 0.000 claims 9
- 239000013604 expression vector Substances 0.000 claims 6
- 238000000034 method Methods 0.000 claims 6
- 108090000623 proteins and genes Proteins 0.000 claims 4
- 102000004169 proteins and genes Human genes 0.000 claims 4
- 101150081864 Spr gene Proteins 0.000 claims 3
- 235000001014 amino acid Nutrition 0.000 claims 3
- 150000001413 amino acids Chemical class 0.000 claims 3
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 claims 2
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 claims 2
- 210000004899 c-terminal region Anatomy 0.000 claims 2
- 239000012636 effector Substances 0.000 claims 2
- 239000001963 growth medium Substances 0.000 claims 2
- 229920001427 mPEG Polymers 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 230000035772 mutation Effects 0.000 claims 2
- -1 poly(ethylene glycol) Polymers 0.000 claims 2
- 235000018102 proteins Nutrition 0.000 claims 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims 1
- 241001646716 Escherichia coli K-12 Species 0.000 claims 1
- 241001302584 Escherichia coli str. K-12 substr. W3110 Species 0.000 claims 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical group OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims 1
- 101150052672 IGS1 gene Proteins 0.000 claims 1
- 108091005804 Peptidases Proteins 0.000 claims 1
- 239000004365 Protease Substances 0.000 claims 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 1
- 235000004279 alanine Nutrition 0.000 claims 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 claims 1
- 125000003277 amino group Chemical group 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- 235000018417 cysteine Nutrition 0.000 claims 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims 1
- 230000000694 effects Effects 0.000 claims 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims 1
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 claims 1
- 210000001322 periplasm Anatomy 0.000 claims 1
- 229920001223 polyethylene glycol Polymers 0.000 claims 1
- 238000011144 upstream manufacturing Methods 0.000 claims 1
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
- 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
-
- 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
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2875—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154
-
- 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
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
-
- 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
- C12N9/90—Isomerases (5.)
-
- 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'
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Claims (15)
1. Rekombinantna gram-negativna bakterijska stanica koja sadrži:
a) ekspresijski vektor koji sadrži rekombinantni polinukleotid koji kodira DsbC; i
b) jedan ili više polinukleotida koji kodiraju antitijelo ili antigen vezujući fragment od njih koji se specifično veže za CD154;
c) mutirani Tsp gen koji kodira Tsp protein koji ima 50% ili manje aktivnosti proteaze divljeg tipa ne-mutiranog Tsp proteina kao što je prikazano u SEK ID BR: 26 i;
d) mutirani spr gen koji kodira spr protein čija je sekvenca divljeg tipa prikazana u SEK ID BR: 24 uključujući
mutaciju koja utječe na aminokiselinu C94; i
pri čemu je navedena gram-negativna bakterijska stanica izabrana od sojeva E. coli K12 i W3110, i pri čemu je bakterijska stanica izogen za navedene sojeve, osim za rekombinantni polinukleotid koji kodira DsbC, jedan ili više polinukleotida koji kodiraju antitijelo ili antigen vezujući fragment od njih koji se specifično veže za CD154 i mutirani Tsp gen i mutirani spr gen.
2. Gram-negativna bakterijska stanica prema patentnom zahtjevu 1, naznačena time da DsbC sadrži histidin–tag na N-terminusu ili C-terminusu.
3. Gram-negativna bakterijska stanica prema patentnom zahtjevu 1 ili 2, naznačena time da ekspresijski vektor sadrži polinukleotid koji ima sekvencu danu u SEK ID BR: 45 ili SEK ID BR: 51.
4. Gram-negativna bakterijska stanica prema bilo kojem od patentnih zahtjeva 1 do 3, naznačena time da antitijelo ili antigen vezujući fragment od njih sadrži varijabilnu domenu teškog lanca koja sadrži tri CDR-a koji imaju sekvencu danu u SEK ID BR: 1 za CDRH1, SEK ID BR: 2, za CDRH2 i SEK ID BR: 3 za CDRH3 i varijabilnu domenu lakog lanca koji sadrži tri CDR-a koji imaju sekvencu danu u SEK ID BR: 4 za CDRL1, SEK ID BR: 5 za CDRL2 i SEK ID BR: 6 za CDRL3.
5. Gram-negativna bakterijska stanica prema patentnom zahtjevu 4, naznačena time da jedan ili više polinukleotida kodiraju antitijelo koje sadrži sekvencu varijabilne regije lakog lanca danu u SEK ID BR: 8 i varijabilnu regiju teškog lanca danu u SEK ID BR: 10.
6. Gram-negativna bakterijska stanica prema bilo kojem od patentnih zahtjeva 1 do 5, naznačena time da je antitijelo Fab ili Fabov fragment.
7. Gram-negativna bakterijska stanica prema patentnom zahtjevu 6, naznačena time da Fab ili Fabov fragment sadrži laki lanac koji ima sekvencu danu u SEK ID BR: 12 i teški lanac, koji ima sekvencu danu u SEK ID BR: 14 ili 16.
8. Gram-negativna bakterijska stanica prema bilo kojem od patentnih zahtjeva 1 do 7, naznačena time da gram-negativna bakterijska stanica sadrži prvi ekspresijski vektor koji sadrži rekombinantni polinukleotid koji kodira DsbC i drugi ekspresijski vektor koji sadrži jedan ili više polinukleotida koji kodiraju antitijelo ili antigen koji veže fragment od njih, posebno koji veže CD154.
9. Gram-negativna bakterijska stanica prema bilo kojem od patentnih zahtjeva 1 do 7, naznačena time da ekspresijski vektor, koji sadrži rekombinantni polinukleotid koji kodira DsbC, dodatno sadrži jedan ili više polinukleotida koji kodiraju antitijelo ili antigen vezujući fragment od njih, posebno koji veže CD154.
10. Gram-negativna bakterijska stanica prema bilo kojem od prethodnih patentnih zahtjeva, naznačena time da mutacija u spr genu koji kodira spr protein, rezultira u promjeni aminokiselinskog cisteina u alanin na poziciji 94 (C94A).
11. Gram-negativna bakterijska stanica prema bilo kojem od patentnih zahtjeva 1 do 10, koji sadrži ekspresijski vektor koji sadrži rekombinantni polinukleotid koji kodira DsbC i dicistronsku poruku za proizvodnju antitijela ili antigen vezujućeg fragmenta od njih, posebno koji veže CD154, u kojem uzvodni cistron sadrži DNA koja kodira laki lanac antitijela i nizvodni cistron sadrži DNA koja kodira odgovarajući teški lanac, naznačena time da dicistronska poruka sadrži sekvencu izabranu od IGS1 (SEK ID BR: 33), IGS2 (SEK ID BR:34), IGS3 (SEK ID BR: 35) i IGS4 (SEK ID BR: 36).
12. Postupak za proizvodnju antitijela ili antigen vezujućeg fragmenta od njih posebno koji veže CD154, naznačen time da sadži:
a) kultiviranje rekombinantne gram-negativne bakterijske stanice kao što je definirano u bilo kojem od patentnih zahtjeva 1 do 11 u mediju kulture pod uvjetima efikasnim da eksprimiraju antitijelo ili antigen vezujući fragment od njih, posebno koji veže CD154 i rekombinantni polinukleotid koji kodira DsbC; i
b) obnavljanje antitijela ili antigen vezujućeg fragmenta od njih, posebno koji veže CD154 iz periplazme rekombinantne gram-negativne bakterijske stanice i/ili medija kulture.
13. Postupak prema patentnom zahtjevu 12, naznačen time da postupak dalje sadrži korak spajanja efektorske molekule na aminokiselinu na ili prema C-terminalnom kraju teškog lanca i/ili lakog lanca antitijela.
14. Postupak prema patentnom zahtjevu 13, naznačen time da efektorska molekula sadrži poli(etilenglikol) ili metoksipoli(etilenglikol).
15. Postupak prema patentnom zahtjevu 14, naznačen time da postupak sadrži vezanje na jedan od cisteinskih ostataka na C-terminalnom kraju teškog lanca lizil-maleimidne skupine, pri čemu svaka amino skupina lizil ostatka ima kovalentno povezan sa njim metoksipoli(etilenglikol) ostatak koji ima molekulsku težinu od oko 20,000 Da.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP11173880 | 2011-07-13 | ||
| PCT/EP2012/002945 WO2013007388A1 (en) | 2011-07-13 | 2012-07-13 | Bacterial host strain expressing recombinant dsbc |
| EP12737492.4A EP2731973B1 (en) | 2011-07-13 | 2012-07-13 | Bacterial host strain expressing recombinant dsbc |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HRP20180226T1 true HRP20180226T1 (hr) | 2018-03-09 |
Family
ID=46545736
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| HRP20180226TT HRP20180226T1 (hr) | 2011-07-13 | 2018-02-06 | Bakterijski soj domaćina koji eksprimira rekombinantnu dsbc |
Country Status (27)
| Country | Link |
|---|---|
| US (2) | US9725516B2 (hr) |
| EP (2) | EP2731973B1 (hr) |
| JP (3) | JP6431370B2 (hr) |
| KR (1) | KR102023786B1 (hr) |
| CN (1) | CN103649124B (hr) |
| AU (1) | AU2012283388B2 (hr) |
| BR (1) | BR112013032871B1 (hr) |
| CA (1) | CA2841824C (hr) |
| CY (1) | CY1119985T1 (hr) |
| DK (1) | DK2731973T3 (hr) |
| EA (1) | EA031449B1 (hr) |
| ES (1) | ES2659155T3 (hr) |
| HK (1) | HK1194393A1 (hr) |
| HR (1) | HRP20180226T1 (hr) |
| HU (1) | HUE035674T2 (hr) |
| IL (1) | IL229947B (hr) |
| IN (1) | IN2014DN00202A (hr) |
| LT (1) | LT2731973T (hr) |
| ME (1) | ME02957B (hr) |
| MX (1) | MX348738B (hr) |
| NO (1) | NO2731973T3 (hr) |
| PL (1) | PL2731973T3 (hr) |
| PT (1) | PT2731973T (hr) |
| RS (1) | RS56926B1 (hr) |
| SG (1) | SG10201807560XA (hr) |
| SI (1) | SI2731973T1 (hr) |
| WO (1) | WO2013007388A1 (hr) |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| MX2009010120A (es) * | 2007-03-22 | 2009-10-19 | Ucb Pharma Sa | Proteinas de union, incluyendo anticuerpos, derivados de anticuerpo y fragmentos de anticuerpo, que se unen especificamente cd154 y sus usos. |
| 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 |
| 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 |
| GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
| GB201012599D0 (en) | 2010-07-27 | 2010-09-08 | Ucb Pharma Sa | Process for purifying proteins |
| 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 |
| GB201223276D0 (en) * | 2012-12-21 | 2013-02-06 | Ucb Pharma Sa | Antibodies and methods of producing same |
| EP4253548A3 (en) | 2014-12-22 | 2024-01-03 | UCB Biopharma SRL | Protein manufacture |
| KR101810778B1 (ko) * | 2015-06-23 | 2017-12-20 | 서울대학교산학협력단 | Cd154 결합 폴리펩타이드 및 그 용도 |
| 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 |
| CN109371049A (zh) * | 2018-11-14 | 2019-02-22 | 天津大学 | 分子伴侣在促进单克隆抗体的形成和/或提高单克隆抗体的表达量中的应用 |
| 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 |
| WO2021023367A1 (de) * | 2019-08-05 | 2021-02-11 | Wacker Chemie Ag | Bakterienstamm zur freisetzung eines rekombinanten proteins in einem fermentationsverfahren |
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| GB0315457D0 (en) | 2003-07-01 | 2003-08-06 | Celltech R&D Ltd | Biological products |
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| GB0425534D0 (en) | 2004-11-19 | 2004-12-22 | Celltech R&D Ltd | Process for obtaining antibodies |
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| 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 |
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| GB201208367D0 (en) | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
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