ME03561B - SOJ BAKTERIJA DOMAĆINA KOJI SADRŽI MUTIRANI SPR GEN I DIVLJI TIP Tsp GENA - Google Patents

SOJ BAKTERIJA DOMAĆINA KOJI SADRŽI MUTIRANI SPR GEN I DIVLJI TIP Tsp GENA

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Publication number
ME03561B
ME03561B MEP-2019-281A MEP2019281A ME03561B ME 03561 B ME03561 B ME 03561B ME P2019281 A MEP2019281 A ME P2019281A ME 03561 B ME03561 B ME 03561B
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mutated
gene
protein
recombinant
cell
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MEP-2019-281A
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German (de)
English (en)
French (fr)
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Mark Ellis
David Paul Humphreys
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Ucb Biopharma Sprl
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    • 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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    • 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
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    • 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
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    • 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
    • C12N1/22—Processes using, or culture media containing, cellulose or hydrolysates thereof
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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; 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
    • C12N1/26—Processes using, or culture media containing, hydrocarbons
    • C12N1/28—Processes using, or culture media containing, hydrocarbons aliphatic
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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
    • 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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    • 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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    • 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
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    • 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'
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts

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  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Description

SOJ BAKTERIJA DOMAĆINA KOJI SADRŽI MUTIRANI SPR GEN I DIVLJI TIP Tsp GENA
Patentni Zahtevi
1. Postupak za proizvodnju rekombinantnog proteina od interesa koji obuhvata kultivaciju rekombinantne gram-negativne bakterijske ćelije u medijumu kulture pod uslovima koji su efikasni za ekspresiju rekombinantnog proteina od interesa i izdvajanje rekombinantnog proteina od interesa iz periplazme rekombinantne gram-negativne bakterijske ćelije i/ili medijuma kulture, naznačen time što rekombinantna gram-negativna bakterijska ćelija sadrži
(a) mutirani spr gen koji kodira mutirani spr protein koji može da suzbije fenotip ćelije koji sadrži mutirani Tsp gen, naznačen time što mutirani spr gen kodira spr protein koji ima mutaciju u aminokiselini H145 prema aminokiselinskoj sekvenci SEK ID NO: 21, i
(b) nerekombinantni divlji tip hromozomskog Tsp gena,
naznačen time što je ćelija izogenična sa divljim tipom ćelije E. coli, osim za mutirani spr gen.
2. Postupak prema patentnom zahtevu 1, naznačen time što mutirani spr gen kodira spr protein koji ima mutaciju H145A, u skladu sa aminokiselinskom sekvencom SEK ID NO: 21.
3. Postupak prema bilo kojem od patentnih zahteva 1 ili 2, naznačen time, što ćelija dalje sadrži i rekombinantni polinukleotid koji kodira DsbC.
4. Postupak prema bilo kojem od patentnih zahteva od 1 do 3, naznačen time što ćelija dalje sadrži jedan ili više od sledećih mutiranih gena:
a) mutirani DegP gen koji kodira DegP protein koji ima aktivnost šaperona i smanjenu proteaznu aktivnost;
b) mutirani ptr gen, pri čemu mutirani ptr gen kodira protein Proteaza III koji ima smanjenu proteaznu aktivnost ili je mutirani nokaut ptr gen; i
c) mutirani OmpT gen, pri čemu mutirani OmpT gen kodira OmpT protein koji ima smanjenu proteaznu aktivnost ili je mutirani nokaut OmpT gen.
5. Postupak prema bilo kojem od patentnih zahteva od 1 do 4, naznačen time što ćelija sadrži vektor koji sadrži rekombinantni polinukleotid koji kodira DsbC i polinukleotidnu sekvencu koja kodira protein od interesa.
6. Postupak prema patentnom zahtevu 5, naznačen time što vektor sadrži promoter koji kontroliše ekspresiju rekombinantnog polinukleotida koji kodira DsbC i polinukleotidnu sekvencu koja kodira protein od interesa.
7. Postupak prema patentnom zahtevu 5 ili 6, naznačen time što je protein od interesa antitelo ili njegov antigen vezujući fragment.
8. Postupak prema patentnom zahtevu 7, naznačen time što je antitelo ili njegov antigen vezujući fragment specifičan za TNF.
9. Postupak prema bilo kojem od patentnih zahteva od 1 do 8, naznačen time što se rekombinantni protein od interesa dobija iz periplazme i/ili supernatanta.
10. Postupak prema patentnom zahtevom 9, naznačen time što sadrži dalji korak prerade proteina od interesa PEGilacijom.
11. Postupak prema bilo kojem od prethodnog patentnog zahteva, naznačen time što je ćelija E. coli W3110.
12. Rekombinantna gram-negativna bakterijska ćelija koja sadrži:
(a) mutirani spr gen koji kodira mutirani spr protein sposoban za supresiju fenotipa ćelije koja sadrži mutirani Tsp gen naznačen time što mutirani spr gen kodira spr protein koji ima mutaciju u aminokiselini H145 prema aminokiselinskoj sekvenci SEK ID NO: 21, i
(b) nerekombinantni hromozomski Tsp gen divljeg tipa,
naznačen time što je ćelija izogenična sa divljim tipom E. coli ćelije osim za mutirani spr gen.
13. Rekombinantna ćelija E. coli W3110 koja sadrži:
(a) mutirani spr gen koji kodira mutirani spr protein koji može da suzbije fenotip ćelije koja sadrži mutirani Tsp gen, pri čemu mutirani spr gen kodira spr protein koji ima mutaciju u aminokiselini H145 prema aminokiselinskoj sekvenci SEK ID NO: 21, i
(b) nerekombinantni hromozomski Tsp gen divljeg tipa.
2

Claims (13)

1. Postupak za proizvodnju rekombinantnog proteina od interesa koji obuhvata kultivaciju rekombinantne gram-negativne bakterijske ćelije u medijumu kulture pod uslovima koji su efikasni za ekspresiju rekombinantnog proteina od interesa i izdvajanje rekombinantnog proteina od interesa iz periplazme rekombinantne gram-negativne bakterijske ćelije i/ili medijuma kulture, naznačen time što rekombinantna gram-negativna bakterijska ćelija sadrži(a) mutirani spr gen koji kodira mutirani spr protein koji može da suzbije fenotip ćelije koji sadrži mutirani Tsp gen, naznačen time što mutirani spr gen kodira spr protein koji ima mutaciju u aminokiselini H145 prema aminokiselinskoj sekvenci SEK ID NO: 21, i(b) nerekombinantni divlji tip hromozomskog Tsp gena, naznačen time što je ćelija izogenična sa divljim tipom ćelije E. coli, osim za mutirani spr gen.
2. Postupak prema patentnom zahtevu 1, naznačen time što mutirani spr gen kodira spr protein koji ima mutaciju H145A, u skladu sa aminokiselinskom sekvencom SEK ID NO: 21.
3. Postupak prema bilo kojem od patentnih zahteva 1 ili 2, naznačen time, što ćelija dalje sadrži i rekombinantni polinukleotid koji kodira DsbC.
4. Postupak prema bilo kojem od patentnih zahteva od 1 do 3, naznačen time što ćelija dalje sadrži jedan ili više od sledećih mutiranih gena:a) mutirani DegP gen koji kodira DegP protein koji ima aktivnost šaperona i smanjenu proteaznu aktivnost;b) mutirani ptr gen, pri čemu mutirani ptr gen kodira protein Proteaza III koji ima smanjenu proteaznu aktivnost ili je mutirani nokaut ptr gen; ic) mutirani OmpT gen, pri čemu mutirani OmpT gen kodira OmpT protein koji ima smanjenu proteaznu aktivnost ili je mutirani nokaut OmpT gen.
5. Postupak prema bilo kojem od patentnih zahteva od 1 do 4, naznačen time što ćelija sadrži vektor koji sadrži rekombinantni polinukleotid koji kodira DsbC i polinukleotidnu sekvencu koja kodira protein od interesa.
6. Postupak prema patentnom zahtevu 5, naznačen time što vektor sadrži promoter koji kontroliše ekspresiju rekombinantnog polinukleotida koji kodira DsbC i polinukleotidnu sekvencu koja kodira protein od interesa.
7. Postupak prema patentnom zahtevu 5 ili 6, naznačen time što je protein od interesa antitelo ili njegov antigen vezujući fragment.
8. Postupak prema patentnom zahtevu 7, naznačen time što je antitelo ili njegov antigen vezujući fragment specifičan za TNF.
9. Postupak prema bilo kojem od patentnih zahteva od 1 do 8, naznačen time što se rekombinantni protein od interesa dobija iz periplazme i/ili supernatanta.
10. Postupak prema patentnom zahtevom 9, naznačen time što sadrži dalji korak prerade proteina od interesa PEGilacijom.
11. Postupak prema bilo kojem od prethodnog patentnog zahteva, naznačen time što je ćelija E. coli W3110.
12. Rekombinantna gram-negativna bakterijska ćelija koja sadrži:(a) mutirani spr gen koji kodira mutirani spr protein sposoban za supresiju fenotipa ćelije koja sadrži mutirani Tsp gen naznačen time što mutirani spr gen kodira spr protein koji ima mutaciju u aminokiselini H145 prema aminokiselinskoj sekvenci SEK ID NO: 21, i(b) nerekombinantni hromozomski Tsp gen divljeg tipa, naznačen time što je ćelija izogenična sa divljim tipom E. coli ćelije osim za mutirani spr gen.
13. Rekombinantna ćelija E. coli W3110 koja sadrži:(a) mutirani spr gen koji kodira mutirani spr protein koji može da suzbije fenotip ćelije koja sadrži mutirani Tsp gen, pri čemu mutirani spr gen kodira spr protein koji ima mutaciju u aminokiselini H145 prema aminokiselinskoj sekvenci SEK ID NO: 21, i(b) nerekombinantni hromozomski Tsp gen divljeg tipa. 2
MEP-2019-281A 2010-01-14 2011-01-13 SOJ BAKTERIJA DOMAĆINA KOJI SADRŽI MUTIRANI SPR GEN I DIVLJI TIP Tsp GENA ME03561B (me)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GBGB1000590.8A GB201000590D0 (en) 2010-01-14 2010-01-14 Bacterial host strain
PCT/EP2011/050415 WO2011086138A1 (en) 2010-01-14 2011-01-13 Bacterial host strain comprising a mutant spr gene and a wild-type tsp gene
EP11700144.6A EP2523975B1 (en) 2010-01-14 2011-01-13 Bacterial host strain comprising a mutant spr gene and a wild-type tsp gene

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ME03561B true ME03561B (me) 2020-07-20

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US (2) US8969037B2 (me)
EP (1) EP2523975B1 (me)
JP (1) JP5960607B2 (me)
KR (1) KR101741865B1 (me)
CN (1) CN102712694B (me)
AU (2) AU2011206586B2 (me)
BR (1) BR112012016807B1 (me)
CA (1) CA2785931C (me)
CY (1) CY1122174T1 (me)
DK (1) DK2523975T3 (me)
ES (1) ES2747981T3 (me)
GB (1) GB201000590D0 (me)
HR (1) HRP20191815T1 (me)
HU (1) HUE046420T2 (me)
LT (1) LT2523975T (me)
ME (1) ME03561B (me)
PL (1) PL2523975T3 (me)
PT (1) PT2523975T (me)
RS (1) RS59417B1 (me)
SG (1) SG182279A1 (me)
SI (1) SI2523975T1 (me)
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