CA3024027A1 - Direct selection of cells expressing high levels of heteromeric proteins using glutamine synthetase intragenic complementation vectors - Google Patents

Direct selection of cells expressing high levels of heteromeric proteins using glutamine synthetase intragenic complementation vectors Download PDF

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Publication number
CA3024027A1
CA3024027A1 CA3024027A CA3024027A CA3024027A1 CA 3024027 A1 CA3024027 A1 CA 3024027A1 CA 3024027 A CA3024027 A CA 3024027A CA 3024027 A CA3024027 A CA 3024027A CA 3024027 A1 CA3024027 A1 CA 3024027A1
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Canada
Prior art keywords
nucleic acid
mutgs
polypeptide
expression system
vector
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CA3024027A
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English (en)
French (fr)
Inventor
Randal R. Ketchem
Jeffrey T. Mcgrew
Dina A. FOMINA YADLIN
Trent P. MUNRO
Neeraj Jagdish Agrawal
Kristine M. DARIS
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Amgen Inc
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Amgen Inc
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Publication of CA3024027A1 publication Critical patent/CA3024027A1/en
Pending legal-status Critical Current

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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
    • C12N15/67—General methods for enhancing the expression
    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1055—Protein x Protein interaction, e.g. two hybrid selection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/06—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies from serum
    • C07K16/065—Purification, fragmentation
    • 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
    • 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
    • C12N15/65—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression using markers
    • 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
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • 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
    • C12N9/93—Ligases (6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y104/00—Oxidoreductases acting on the CH-NH2 group of donors (1.4)
    • C12Y104/01—Oxidoreductases acting on the CH-NH2 group of donors (1.4) with NAD+ or NADP+ as acceptor (1.4.1)
    • C12Y104/01002—Glutamate dehydrogenase (1.4.1.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y603/00—Ligases forming carbon-nitrogen bonds (6.3)
    • C12Y603/04—Other carbon-nitrogen ligases (6.3.4)
    • C12Y603/04004—Adenylosuccinate synthase (6.3.4.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y603/00—Ligases forming carbon-nitrogen bonds (6.3)
    • C12Y603/04—Other carbon-nitrogen ligases (6.3.4)
    • C12Y603/04005—Argininosuccinate synthase (6.3.4.5)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
    • 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
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
    • C12N2015/8518—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic expressing industrially exogenous proteins, e.g. for pharmaceutical use, human insulin, blood factors, immunoglobulins, pseudoparticles
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y603/00—Ligases forming carbon-nitrogen bonds (6.3)
    • C12Y603/01—Acid-ammonia (or amine)ligases (amide synthases)(6.3.1)
    • C12Y603/01002—Glutamate-ammonia ligase (6.3.1.2)

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
CA3024027A 2016-05-11 2017-05-11 Direct selection of cells expressing high levels of heteromeric proteins using glutamine synthetase intragenic complementation vectors Pending CA3024027A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201662334966P 2016-05-11 2016-05-11
US62/334,966 2016-05-11
PCT/US2017/032139 WO2017197098A1 (en) 2016-05-11 2017-05-11 Direct selection of cells expressing high levels of heteromeric proteins using glutamine synthetase intragenic complementation vectors

Publications (1)

Publication Number Publication Date
CA3024027A1 true CA3024027A1 (en) 2017-11-16

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Family Applications (1)

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CA3024027A Pending CA3024027A1 (en) 2016-05-11 2017-05-11 Direct selection of cells expressing high levels of heteromeric proteins using glutamine synthetase intragenic complementation vectors

Country Status (17)

Country Link
US (2) US11384140B2 (https=)
EP (2) EP3455359B1 (https=)
JP (2) JP6972022B2 (https=)
KR (2) KR102439719B1 (https=)
CN (2) CN115873904A (https=)
AU (2) AU2017263454B2 (https=)
BR (1) BR112018073316A2 (https=)
CA (1) CA3024027A1 (https=)
CL (1) CL2018003210A1 (https=)
EA (1) EA201892588A1 (https=)
ES (1) ES2914118T3 (https=)
IL (2) IL300591A (https=)
MA (1) MA44976A (https=)
MX (2) MX394682B (https=)
SG (1) SG11201810040WA (https=)
TW (2) TWI757291B (https=)
WO (1) WO2017197098A1 (https=)

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US20250101402A1 (en) * 2021-08-03 2025-03-27 Shanghai Zhenge Biotechnology Co., Ltd. Selectable markers for eukaryotic expression system
WO2023125389A1 (en) * 2021-12-27 2023-07-06 Shanghai Zhenge Biotechnology Co., Ltd. Novel markers for recombinant production system
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AU2023237597A1 (en) 2022-03-23 2024-10-31 Boehringer Ingelheim International Gmbh New glutamine synthetase variants as selection marker
WO2025037708A1 (en) * 2023-08-16 2025-02-20 Korea Advanced Institute Of Science And Technology Method for selection of cell line expressing high levels of recombinant protein using glutamine synthetase split expression vector
WO2025128343A1 (en) 2023-12-11 2025-06-19 Just-Evotec Biologics, Inc. Protein expression using trans-splicing and split selectable markers
CN118006685B (zh) * 2024-04-07 2024-07-23 上海碧博生物医药科技有限公司 一种快速的高表达单克隆细胞株构建方法
KR20250169382A (ko) * 2024-05-23 2025-12-03 한국과학기술원 디히드로엽산 환원효소 분할 발현벡터를 이용한 재조합 단백질 고발현 세포주의 선별방법
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Also Published As

Publication number Publication date
SG11201810040WA (en) 2018-12-28
MX394682B (es) 2025-03-24
KR102439719B1 (ko) 2022-09-02
IL300591A (en) 2023-04-01
IL262923A (en) 2019-01-31
KR102323519B1 (ko) 2021-11-09
ES2914118T3 (es) 2022-06-07
WO2017197098A1 (en) 2017-11-16
AU2017263454B2 (en) 2023-02-09
US20230048658A1 (en) 2023-02-16
AU2017263454A1 (en) 2018-12-06
TWI821905B (zh) 2023-11-11
TW202223093A (zh) 2022-06-16
CN109219660A (zh) 2019-01-15
EP3455359B1 (en) 2022-03-09
MX2022009674A (es) 2022-09-09
US11384140B2 (en) 2022-07-12
CN115873904A (zh) 2023-03-31
TW201805422A (zh) 2018-02-16
US20190127452A1 (en) 2019-05-02
CN109219660B (zh) 2023-01-06
AU2023200660B2 (en) 2024-12-12
IL262923B2 (en) 2023-08-01
JP7344949B2 (ja) 2023-09-14
TWI757291B (zh) 2022-03-11
MX2018013882A (es) 2019-03-14
JP2022033734A (ja) 2022-03-02
KR20210135635A (ko) 2021-11-15
US12516111B2 (en) 2026-01-06
JP2019514417A (ja) 2019-06-06
JP6972022B2 (ja) 2021-11-24
AU2023200660A1 (en) 2023-03-09
BR112018073316A2 (pt) 2019-06-04
MA44976A (fr) 2019-03-20
IL262923B1 (en) 2023-04-01
EP4067493A1 (en) 2022-10-05
CL2018003210A1 (es) 2019-02-15
KR20190005193A (ko) 2019-01-15
EA201892588A1 (ru) 2019-06-28
EP3455359A1 (en) 2019-03-20

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