HRP20231139T1 - Proizvodnja heteromultimernih proteina upotrebom stanica sisavaca - Google Patents

Proizvodnja heteromultimernih proteina upotrebom stanica sisavaca Download PDF

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Publication number
HRP20231139T1
HRP20231139T1 HRP20231139TT HRP20231139T HRP20231139T1 HR P20231139 T1 HRP20231139 T1 HR P20231139T1 HR P20231139T T HRP20231139T T HR P20231139TT HR P20231139 T HRP20231139 T HR P20231139T HR P20231139 T1 HRP20231139 T1 HR P20231139T1
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Croatia
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host cell
combined culture
cell
heavy chain
combined
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HRP20231139TT
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Justin Scheer
Whitney SHATZ
Domingos Ng
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F. Hoffmann - La Roche Ag
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Publication of HRP20231139T1 publication Critical patent/HRP20231139T1/hr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/02—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length in solution
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/02—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length in solution
    • C07K1/026—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length in solution by fragment condensation in solution
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins
    • C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • 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
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/526—CH3 domain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/53—Hinge
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • 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
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Analytical Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Epidemiology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Mycology (AREA)

Claims (6)

1. Postupak proizvodnje bispecifičnog protutijela koje obuhvaća i) prvi teški lanac protutijela koji sadrži modifikaciju čvora, koja uključuje supstituciju T366W (EU numeracija), pri čemu je prvi teški lanac protutijela povezan s prvim lakim lancem, i ii) drugi teški lanac protutijela koji uključuje modifikaciju šupljine, koja uključuje dvije ili više supstitucija aminokiselina odabranih iz skupine koja se sastoji od T366S, L368A i Y407V (EU numeracija), pri čemu je drugi teški lanac protutijela povezan s drugim lakim lancem, pri čemu modifikacija šupljine stupa u interakciju s modifikacijom čvora na sučelju, i pri čemu su prvi i drugi teški lanac protutijela povezani najmanje jednom međulančanom disulfidnom vezom, a postupak uključuje sljedeće korake: (a) uzgoj kombinirane kulture prve stanice domaćina i druge stanice domaćina, pri čemu prva stanica domaćina eksprimira prvi teški lanac protutijela i prvi laki lanac, pri čemu druga stanica domaćina eksprimira drugi teški lanac protutijela i drugi laki lanac, i pri čemu su prva stanica domaćina i druga stanica domaćina obje stanica sisavca; (b) dodavanje reduciranog glutationa (GSH) u kombiniranu kulturu kako bi se postigla konačna koncentracija od 5 mM do 20 mM; i (c) prikupljanje kombiniranog medija kulture iz kombinirane kulture bez razaranja stanične membrane 4 do 24 sata nakon dodavanja GSH u kombiniranu kulturu, pri čemu prikupljanje uključuje uklanjanje prve stanice domaćina i druge stanice domaćina iz kombiniranog medija kulture, dok kombinirani medij kulture sadrži bispecifično protutijelo, i gdje je bispecifično protutijelo ljudsko ili humanizirano.
2. Postupak prema patentnom zahtjevu 1, naznačen time, da se kombinirani medij kulture sakuplja 15 sati nakon dodavanja GSH.
3. Postupak prema patentnom zahtjevu 1 ili 2, naznačen time, da se kombinirani medij kulture inkubira 4 sata do 7 dana.
4. Postupak prema bilo kojem od patentnih zahtjeva 1 do 3, naznačen time, da se prva stanica domaćina i druga stanica domaćina uzgajaju odvojeno prije spajanja u kombiniranu kulturu.
5. Postupak prema patentnom zahtjevu 4, naznačen time, da se GSH dodaje u kombiniranu staničnu kulturu kako bi se postigla konačna koncentracija bilo koje od 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM ili 20 mM; poželjno pri čemu je konačna koncentracija 15 mM.
6. Postupak prema bilo kojem od patentnih zahtjeva 1 do 5, naznačen time, da je prva i/ili druga stanica domaćina a) CHO stanica; i/ili b) stabilna stanična linija.
HRP20231139TT 2014-05-06 2015-05-06 Proizvodnja heteromultimernih proteina upotrebom stanica sisavaca HRP20231139T1 (hr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201461989509P 2014-05-06 2014-05-06
EP15789583.0A EP3140392B1 (en) 2014-05-06 2015-05-06 Production of heteromultimeric proteins using mammalian cells
PCT/US2015/029546 WO2015171822A1 (en) 2014-05-06 2015-05-06 Production of heteromultimeric proteins using mammalian cells

Publications (1)

Publication Number Publication Date
HRP20231139T1 true HRP20231139T1 (hr) 2024-01-05

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HRP20231139TT HRP20231139T1 (hr) 2014-05-06 2015-05-06 Proizvodnja heteromultimernih proteina upotrebom stanica sisavaca

Country Status (14)

Country Link
US (1) US10941190B2 (hr)
EP (2) EP4306544A3 (hr)
JP (3) JP6715186B2 (hr)
KR (2) KR20230164192A (hr)
CN (1) CN106471117A (hr)
BR (2) BR112016024462B1 (hr)
CA (1) CA2943707A1 (hr)
ES (1) ES2955736T3 (hr)
HR (1) HRP20231139T1 (hr)
HU (1) HUE063273T2 (hr)
MX (2) MX2016014409A (hr)
PL (1) PL3140392T3 (hr)
RU (1) RU2727012C2 (hr)
WO (1) WO2015171822A1 (hr)

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TW201834685A (zh) * 2016-12-29 2018-10-01 財團法人生物技術開發中心 Klk6-介導cns-特異性抗體前藥活化
TW202016151A (zh) 2018-06-09 2020-05-01 德商百靈佳殷格翰國際股份有限公司 針對癌症治療之多特異性結合蛋白
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