EA201391282A1 - Клеточные линии с низким содержанием фукозы и их применение - Google Patents

Клеточные линии с низким содержанием фукозы и их применение

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Publication number
EA201391282A1
EA201391282A1 EA201391282A EA201391282A EA201391282A1 EA 201391282 A1 EA201391282 A1 EA 201391282A1 EA 201391282 A EA201391282 A EA 201391282A EA 201391282 A EA201391282 A EA 201391282A EA 201391282 A1 EA201391282 A1 EA 201391282A1
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EA
Eurasian Patent Office
Prior art keywords
cells
fucose
population
expression
recombinant proteins
Prior art date
Application number
EA201391282A
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English (en)
Other versions
EA032513B1 (ru
Inventor
Дэниел Хельман
Мейрав Бар-Шимон
Мира Тойстер-Ачитув
Original Assignee
Мерк Сероно С.А.
Priority date (The priority date 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 date listed.)
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Publication date
Priority claimed from IL211583A external-priority patent/IL211583A0/en
Priority claimed from IL217216A external-priority patent/IL217216A0/en
Application filed by Мерк Сероно С.А. filed Critical Мерк Сероно С.А.
Publication of EA201391282A1 publication Critical patent/EA201391282A1/ru
Publication of EA032513B1 publication Critical patent/EA032513B1/ru

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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
    • 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
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • 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
    • C12N15/01—Preparation of mutants without inserting foreign genetic material therein; Screening processes 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
    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1048—Glycosyltransferases (2.4)
    • C12N9/1051—Hexosyltransferases (2.4.1)
    • 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
    • C12N9/88—Lyases (4.)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01271—GDP-L-fucose synthase (1.1.1.271)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y204/00—Glycosyltransferases (2.4)
    • C12Y204/01—Hexosyltransferases (2.4.1)
    • C12Y204/01068—Glycoprotein 6-alpha-L-fucosyltransferase (2.4.1.68), i.e. FUT8
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y402/00—Carbon-oxygen lyases (4.2)
    • C12Y402/01—Hydro-lyases (4.2.1)
    • C12Y402/01047—GDP-mannose 4,6-dehydratase (4.2.1.47), i.e. GMD
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
    • C07K2317/41—Glycosylation, sialylation, or fucosylation
    • 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

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Описан способ отбора клеток с нулевым уровнем фукозы, применимых в качестве клеток-хозяев для экспрессии рекомбинантных белков. Этот способ предусматривает: 1) введение генетических мутаций в популяцию клеток СНО путем контактирования этих клеток с метотрексатом (МТХ), 2) контактирование указанной популяции клеток СНО, содержащей мутантные клетки, с нетоксичным веществом, связывающим фукозу, в течение времени, которое дает возможность осуществить связывание вещества, связывающего фукозу, с фукозной группой на клеточной мембране популяции клеток, где период времени не дает возможности осуществить уничтожение клеток; и 3) извлечение из этой популяции клеток, содержащей мутантные клетки, субпопуляции клеток, которые связываются с веществом, связывающим фукозу, таким образом, отбирая клетки, применимые в качестве клеток-хозяев для экспрессии рекомбинантных белков, отобранные клетки имеют нулевое содержание фукозы. Также описаны клетки и клеточные линии, применимые в качестве клеток-хозяев для экспрессии рекомбинантных белков.
EA201391282A 2011-03-06 2012-03-05 Клеточные линии с низким содержанием фукозы и их применение EA032513B1 (ru)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
IL211583A IL211583A0 (en) 2011-03-06 2011-03-06 Low fucose cell lines and uses thereof
IL217216A IL217216A0 (en) 2011-12-27 2011-12-27 Low fucose cell lins and uses thereof
PCT/IL2012/000109 WO2012120500A2 (en) 2011-03-06 2012-03-05 Low fucose cell lines and uses thereof

Publications (2)

Publication Number Publication Date
EA201391282A1 true EA201391282A1 (ru) 2014-04-30
EA032513B1 EA032513B1 (ru) 2019-06-28

Family

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EA201391282A EA032513B1 (ru) 2011-03-06 2012-03-05 Клеточные линии с низким содержанием фукозы и их применение

Country Status (14)

Country Link
US (1) US9574003B2 (ru)
EP (1) EP2683821B1 (ru)
JP (2) JP2014509850A (ru)
CN (2) CN103597073B (ru)
AU (1) AU2012226398B9 (ru)
BR (1) BR112013022832A2 (ru)
CA (1) CA2829110C (ru)
EA (1) EA032513B1 (ru)
ES (1) ES2687771T3 (ru)
IL (1) IL228184B (ru)
MX (1) MX346663B (ru)
SG (1) SG192632A1 (ru)
WO (1) WO2012120500A2 (ru)
ZA (1) ZA201306336B (ru)

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Also Published As

Publication number Publication date
AU2012226398B9 (en) 2017-04-13
CN103597073A (zh) 2014-02-19
JP2014509850A (ja) 2014-04-24
ES2687771T3 (es) 2018-10-29
AU2012226398B2 (en) 2017-03-23
AU2012226398A1 (en) 2013-09-12
EP2683821A2 (en) 2014-01-15
CN103597073B (zh) 2019-06-07
US9574003B2 (en) 2017-02-21
CA2829110C (en) 2019-01-15
SG192632A1 (en) 2013-09-30
JP2017079775A (ja) 2017-05-18
BR112013022832A2 (pt) 2016-11-22
EP2683821B1 (en) 2018-07-25
WO2012120500A3 (en) 2012-11-15
ZA201306336B (en) 2014-10-29
CN109797138A (zh) 2019-05-24
CA2829110A1 (en) 2012-09-13
IL228184B (en) 2018-12-31
EA032513B1 (ru) 2019-06-28
WO2012120500A2 (en) 2012-09-13
US20140005368A1 (en) 2014-01-02
MX346663B (es) 2017-03-27
JP6353023B2 (ja) 2018-07-04
MX2013010210A (es) 2013-09-26

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