DE60233738D1 - Hemmung des apoptoseprozesses und verbesserung der zelleistung - Google Patents

Hemmung des apoptoseprozesses und verbesserung der zelleistung

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Publication number
DE60233738D1
DE60233738D1 DE60233738T DE60233738T DE60233738D1 DE 60233738 D1 DE60233738 D1 DE 60233738D1 DE 60233738 T DE60233738 T DE 60233738T DE 60233738 T DE60233738 T DE 60233738T DE 60233738 D1 DE60233738 D1 DE 60233738D1
Authority
DE
Germany
Prior art keywords
cell
apoptose
inhibition
improvement
cell performance
Prior art date
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.)
Expired - Lifetime
Application number
DE60233738T
Other languages
English (en)
Inventor
Mitchell Reff
Michael J Betenbaugh
Bruno Figueroa Jr
Eric Ailor
Marie Hardwick
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Biogen Inc
Johns Hopkins University
Original Assignee
Johns Hopkins University
Biogen Idec Inc
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.)
Filing date
Publication date
Application filed by Johns Hopkins University, Biogen Idec Inc filed Critical Johns Hopkins University
Application granted granted Critical
Publication of DE60233738D1 publication Critical patent/DE60233738D1/de
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/10Cells modified by introduction of foreign genetic material
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4747Apoptosis related proteins
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K39/395Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/005Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/721Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K16/00Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2875Immunoglobulins [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
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/67General methods for enhancing the expression
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2800/00Nucleic acids vectors
    • C12N2800/10Plasmid DNA
    • C12N2800/108Plasmid DNA episomal vectors
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2830/00Vector systems having a special element relevant for transcription
    • C12N2830/001Vector systems having a special element relevant for transcription controllable enhancer/promoter combination
    • C12N2830/002Vector systems having a special element relevant for transcription controllable enhancer/promoter combination inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2830/00Vector systems having a special element relevant for transcription
    • C12N2830/15Vector systems having a special element relevant for transcription chimeric enhancer/promoter combination
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2840/00Vectors comprising a special translation-regulating system
    • C12N2840/20Vectors comprising a special translation-regulating system translation of more than one cistron
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2840/00Vectors comprising a special translation-regulating system
    • C12N2840/44Vectors comprising a special translation-regulating system being a specific part of the splice mechanism, e.g. donor, acceptor

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Public Health (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Cell Biology (AREA)
  • Endocrinology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Mycology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
DE60233738T 2001-07-10 2002-07-10 Hemmung des apoptoseprozesses und verbesserung der zelleistung Expired - Lifetime DE60233738D1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US30381301P 2001-07-10 2001-07-10
PCT/US2002/021606 WO2003006607A2 (en) 2001-07-10 2002-07-10 Inhibition of apoptosis process and improvement of cell performance

Publications (1)

Publication Number Publication Date
DE60233738D1 true DE60233738D1 (de) 2009-10-29

Family

ID=23173824

Family Applications (1)

Application Number Title Priority Date Filing Date
DE60233738T Expired - Lifetime DE60233738D1 (de) 2001-07-10 2002-07-10 Hemmung des apoptoseprozesses und verbesserung der zelleistung

Country Status (13)

Country Link
US (1) US7604989B2 (de)
EP (1) EP1414948B1 (de)
JP (2) JP4435563B2 (de)
KR (1) KR20040044433A (de)
CN (1) CN1526011A (de)
AT (1) ATE443131T1 (de)
AU (1) AU2002315534B2 (de)
CA (1) CA2452005C (de)
DE (1) DE60233738D1 (de)
MX (1) MXPA04000281A (de)
NO (1) NO20040101L (de)
WO (1) WO2003006607A2 (de)
ZA (2) ZA200508985B (de)

Families Citing this family (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7812148B2 (en) 2001-04-05 2010-10-12 Millipore Corporation Vectors comprising CpG islands without position effect varigation and having increased expression
KR100454016B1 (ko) * 2002-01-05 2004-10-26 한국과학기술원 항예정사 유전자로 형질전환되고 dhfr 유전자가결핍된 신규한 cho 세포주, 그의 제조 방법 및 상기형질전환된 cho 숙주 세포를 이용한 목적단백질의 생산방법
CA2514412A1 (en) 2003-02-01 2004-08-12 Innovata Plc Improved genetic elements providing high levels of expression
JP4825518B2 (ja) * 2003-10-24 2011-11-30 一般財団法人化学及血清療法研究所 新規なタンパク質高産生組換え動物細胞、その作製方法及びそれを用いたタンパク質を大量産生する方法
EP1702071B1 (de) * 2003-12-31 2012-02-22 Kalobios Inc. Transaktivierungssystem für säugerzellen
US7608425B2 (en) * 2004-07-23 2009-10-27 Immunomedics, Inc. Methods for protein expression in mammalian cells in serum-free medium
US7537930B2 (en) 2004-07-23 2009-05-26 Immunomedics, Inc. Mammalian cell lines for increasing longevity and protein yield from a cell culture
AU2005333513B2 (en) * 2004-07-23 2011-06-02 Immunomedics, Inc. Methods and compositions for increasing longevity and protein yield from a cell culture
WO2007015691A2 (en) * 2004-07-23 2007-02-08 Immunomedics, Inc. Methods and compositions for increasing longevity and protein yield from a cell culture
US7531327B2 (en) * 2004-07-23 2009-05-12 Immunomedics, Inc. Methods and compositions for increasing longevity and protein yield from a cell culture
GB0509965D0 (en) 2005-05-17 2005-06-22 Ml Lab Plc Improved expression elements
WO2007119774A1 (ja) * 2006-04-13 2007-10-25 Chugai Seiyaku Kabushiki Kaisha タウリントランスポーター遺伝子
WO2007124106A2 (en) * 2006-04-21 2007-11-01 Bayer Healthcare Llc Application of antl-apoptotic genes in mammalian cells for perfusion culture.
CN101802170A (zh) * 2007-01-08 2010-08-11 米利波尔公司 在存在降低水平的一或多种污染物的条件下制备重组蛋白的细胞培养方法
CA2727010C (en) 2008-06-13 2017-05-16 Centocor Ortho Biotech Inc. Methods for improving viability and productivity in cell culture
WO2011056886A2 (en) * 2009-11-03 2011-05-12 The Johns Hopkins University Method and composition for generating programmed cell death resistant algal cells
HUP1000243A2 (en) 2010-05-06 2012-01-30 Avidin Kft 8-hidroxy-quinoline derivatives
KR20200002050A (ko) 2018-06-28 2020-01-08 삼성디스플레이 주식회사 유기 발광 표시 장치

Family Cites Families (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6136310A (en) * 1991-07-25 2000-10-24 Idec Pharmaceuticals Corporation Recombinant anti-CD4 antibodies for human therapy
ATE236987T1 (de) * 1992-11-13 2003-04-15 Idec Pharma Corp Konsensus-kozak-sequenzen zur säugetier- exprimierung
US5565491A (en) * 1994-01-31 1996-10-15 Bristol-Myers Squibb Company Use of phosphotyrosine phospatase inhibitors for controlling cellular proliferation
US5550050A (en) * 1994-04-15 1996-08-27 Cytotherapeutics, Inc. Method for implanting encapsulated cells in a host
US5811524A (en) * 1995-06-07 1998-09-22 Idec Pharmaceuticals Corporation Neutralizing high affinity human monoclonal antibodies specific to RSV F-protein and methods for their manufacture and therapeutic use thereof
US6001358A (en) * 1995-11-07 1999-12-14 Idec Pharmaceuticals Corporation Humanized antibodies to human gp39, compositions containing thereof
US6087129A (en) * 1996-01-19 2000-07-11 Betagene, Inc. Recombinant expression of proteins from secretory cell lines
US5897992A (en) * 1996-09-27 1999-04-27 Cold Spring Harbor Laboratory Cell-free assay using oncogene-induced apoptosis in drug-resistant cells
US6011138A (en) * 1997-02-20 2000-01-04 Idec Pharmaceuticals Corporation Gamma-1 anti-human CD23 monoclonal antibodies
US5969102A (en) * 1997-03-03 1999-10-19 St. Jude Children's Research Hospital Lymphocyte surface receptor that binds CAML, nucleic acids encoding the same and methods of use thereof
US5830698A (en) * 1997-03-14 1998-11-03 Idec Pharmaceuticals Corporation Method for integrating genes at specific sites in mammalian cells via homologous recombination and vectors for accomplishing the same
NZ510357A (en) * 1998-08-07 2004-02-27 Immunex Corp Molecules designated B7L-1
US6570002B1 (en) 1999-02-26 2003-05-27 The Johns Hopkins University Inhibitor of programmed cell death
US6140118A (en) * 1999-08-11 2000-10-31 Wisconsin Alumni Research Foundation Avian blastodermal cell lines
EP1650307B1 (de) * 1999-09-27 2010-07-14 Genentech, Inc. Verfahren zur Herstellung von rekombinanten Proteinen mittels Inhibitoren von Apoptose
AU781552B2 (en) * 1999-11-12 2005-05-26 Merck Sharp & Dohme Corp. Diaryl piperidyl pyrrole derivatives as antiprotozoal agents
KR100454016B1 (ko) 2002-01-05 2004-10-26 한국과학기술원 항예정사 유전자로 형질전환되고 dhfr 유전자가결핍된 신규한 cho 세포주, 그의 제조 방법 및 상기형질전환된 cho 숙주 세포를 이용한 목적단백질의 생산방법
EP1348758A1 (de) 2002-03-28 2003-10-01 Boehringer Ingelheim Pharma GmbH & Co.KG Wirtszellen mit verbessertem Überleben und Verfahren zur Herstellung von diesen Zellen

Also Published As

Publication number Publication date
EP1414948A4 (de) 2006-06-21
AU2002315534B2 (en) 2008-05-22
US7604989B2 (en) 2009-10-20
JP2005525782A (ja) 2005-09-02
ZA200508985B (en) 2006-07-26
ATE443131T1 (de) 2009-10-15
WO2003006607A2 (en) 2003-01-23
JP2009118851A (ja) 2009-06-04
ZA200400137B (en) 2006-06-28
CA2452005C (en) 2011-03-15
WO2003006607A9 (en) 2003-04-10
CN1526011A (zh) 2004-09-01
JP4435563B2 (ja) 2010-03-17
NO20040101L (no) 2004-03-09
MXPA04000281A (es) 2004-07-23
EP1414948B1 (de) 2009-09-16
EP1414948A2 (de) 2004-05-06
WO2003006607A3 (en) 2003-07-31
CA2452005A1 (en) 2003-01-23
US20030064510A1 (en) 2003-04-03
KR20040044433A (ko) 2004-05-28

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