WO2010005227A2 - 동물세포용 재조합 발현벡터 - Google Patents

동물세포용 재조합 발현벡터 Download PDF

Info

Publication number
WO2010005227A2
WO2010005227A2 PCT/KR2009/003714 KR2009003714W WO2010005227A2 WO 2010005227 A2 WO2010005227 A2 WO 2010005227A2 KR 2009003714 W KR2009003714 W KR 2009003714W WO 2010005227 A2 WO2010005227 A2 WO 2010005227A2
Authority
WO
WIPO (PCT)
Prior art keywords
cell line
animal cell
dhfr
vector
present
Prior art date
Application number
PCT/KR2009/003714
Other languages
English (en)
French (fr)
Other versions
WO2010005227A3 (ko
WO2010005227A8 (ko
Inventor
김근수
주혜경
전춘주
김재섭
Original Assignee
제넥셀세인 주식회사
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 제넥셀세인 주식회사 filed Critical 제넥셀세인 주식회사
Priority to EP09794633.9A priority Critical patent/EP2314701B1/en
Priority to BRPI0915734-4A priority patent/BRPI0915734A2/pt
Priority to JP2011517345A priority patent/JP5514818B2/ja
Priority to AU2009270108A priority patent/AU2009270108B2/en
Priority to CA2733119A priority patent/CA2733119C/en
Priority to CN200980126484.1A priority patent/CN102177239B/zh
Priority to US13/002,814 priority patent/US8426570B2/en
Publication of WO2010005227A2 publication Critical patent/WO2010005227A2/ko
Publication of WO2010005227A3 publication Critical patent/WO2010005227A3/ko
Publication of WO2010005227A8 publication Critical patent/WO2010005227A8/ko

Links

Classifications

    • 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
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • 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
    • 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
    • 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
    • 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.)
    • C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
    • C12N9/0026—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on CH-NH groups of donors (1.5)
    • C12N9/0028—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on CH-NH groups of donors (1.5) with NAD or NADP as acceptor (1.5.1)
    • C12N9/003—Dihydrofolate reductase [DHFR] (1.5.1.3)
    • 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
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/10—Plasmid DNA
    • C12N2800/106—Plasmid DNA for vertebrates
    • C12N2800/107—Plasmid DNA for vertebrates for mammalian

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

본 발명은 다이하이드로폴레이트 환원효소(Dihydrofolate reductase:DHFR) 프로모터에 작동적으로 연결된(operatively linked) DHFR-코딩 뉴클레오타이드 서열을 포함하는 동물세포용 재조합 발현벡터 및 상기 벡터에 의해 형질전환된 동물세포주, 그리고 그를 이용한 목적단백질의 제조방법에 관한 것이다. 본 발명의 벡터는 기존 동물세포 발현벡터와 비교하여 훨씬 적은 메토트렉세이트 농도 하에서도 효과적으로 DHFR 유전자와 함께 외래 유전자가 증폭된 세포주 클론을 선별할 수 있도록 한다. 또한 본 발명에 따르면, 단백질 생산 세포주 확립과정에서 감소된 농도의 메토드렉세이트의 사용을 통해서 높은 생산성의 세포주를 단기간에 확보하게 되므로 세포주 제작 측면에서 우수한 효과를 나타낸다.
PCT/KR2009/003714 2008-07-10 2009-07-07 동물세포용 재조합 발현벡터 WO2010005227A2 (ko)

Priority Applications (7)

Application Number Priority Date Filing Date Title
EP09794633.9A EP2314701B1 (en) 2008-07-10 2009-07-07 Recombinant expression vector for animal cell
BRPI0915734-4A BRPI0915734A2 (pt) 2008-07-10 2009-07-07 Vetor de expressão recombinante para células de animais
JP2011517345A JP5514818B2 (ja) 2008-07-10 2009-07-07 動物細胞用組み換え発現ベクター
AU2009270108A AU2009270108B2 (en) 2008-07-10 2009-07-07 Recombinant expression vector for animal cell
CA2733119A CA2733119C (en) 2008-07-10 2009-07-07 Recombinant expression vector for animal cell
CN200980126484.1A CN102177239B (zh) 2008-07-10 2009-07-07 用于动物细胞的重组表达载体
US13/002,814 US8426570B2 (en) 2008-07-10 2009-07-07 Recombinant expression vector for animal cell

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020080066771A KR101030978B1 (ko) 2008-07-10 2008-07-10 동물세포용 재조합 발현벡터
KR10-2008-0066771 2008-07-10

Publications (3)

Publication Number Publication Date
WO2010005227A2 true WO2010005227A2 (ko) 2010-01-14
WO2010005227A3 WO2010005227A3 (ko) 2010-04-22
WO2010005227A8 WO2010005227A8 (ko) 2010-06-24

Family

ID=41507570

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2009/003714 WO2010005227A2 (ko) 2008-07-10 2009-07-07 동물세포용 재조합 발현벡터

Country Status (9)

Country Link
US (1) US8426570B2 (ko)
EP (1) EP2314701B1 (ko)
JP (1) JP5514818B2 (ko)
KR (1) KR101030978B1 (ko)
CN (1) CN102177239B (ko)
AU (1) AU2009270108B2 (ko)
BR (1) BRPI0915734A2 (ko)
CA (1) CA2733119C (ko)
WO (1) WO2010005227A2 (ko)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013544080A (ja) * 2010-10-26 2013-12-12 ハンミ サイエンス カンパニー リミテッド ヒト血液凝固第ｖｉｉ因子の大量生産方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10196601B2 (en) 2013-08-20 2019-02-05 Lek Pharmaceuticals D.D. Cell culture medium and process for controlling α-amidation and/or C-terminal amino acid cleavage of polypeptides

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4656134A (en) 1982-01-11 1987-04-07 Board Of Trustees Of Leland Stanford Jr. University Gene amplification in eukaryotic cells

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5011795A (en) 1983-01-19 1991-04-30 Genentech, Inc. Human tPA production using vectors coding for DHFR protein
EP0281246A3 (en) * 1987-02-04 1989-07-12 Invitron Corporation Method to enhance amplification and expression of foreign genes
EP0555203B1 (en) * 1988-12-08 1995-10-18 Abbott Laboratories Expression induction method employing dhfr gene
US5736137A (en) * 1992-11-13 1998-04-07 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma
CN1117155C (zh) * 1994-07-29 2003-08-06 史密丝克莱恩比彻姆有限公司 新型化合物
EP1694850B1 (en) * 2003-11-12 2011-06-29 Schering Corporation Plasmid system for multigene expression
US7846724B2 (en) 2006-04-11 2010-12-07 Hoffmann-La Roche Inc. Method for selecting CHO cell for production of glycosylated antibodies
KR100880509B1 (ko) 2006-10-16 2009-01-28 한미약품 주식회사 재조합 단백질의 대량 생산을 위한 신규한 벡터, 발현세포주 및 이를 이용한 재조합 단백질의 생산 방법
CN101687933B (zh) * 2007-05-30 2015-11-25 浦项工科大学校产学协力团 免疫球蛋白融合蛋白

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4656134A (en) 1982-01-11 1987-04-07 Board Of Trustees Of Leland Stanford Jr. University Gene amplification in eukaryotic cells

Non-Patent Citations (31)

* Cited by examiner, † Cited by third party
Title
ALT ET AL.: "Symp Quant Biol.", vol. 2, 1978, COLD SPRING HARB, pages: 649 - 57
BAI ET AL., ZHONGHUA YI XUE ZA ZHI. FEB, vol. 25;83, no. 4, 2003, pages 333 - 7
BARNES, D. ET AL., ANAL. BIOCHEM., vol. 102, 1980, pages 255
BATT, D. B; G. G. CARMICHAEL, MOL. CELL. BIOL., vol. 15, 1995, pages 4783 - 4790
BIOSCIENCE AND BIOTECHNOLOGY, 11 March 2008 (2008-03-11)
CAPECCHI, M.R., CELL, vol. 22, 1980, pages 479
COLE, C. N.; T. P. STACY, MOL. CELL. BIOL., vol. 5, 1985, pages 2104 - 2113
DULBECCO, R. ET AL., VIROLOGY, vol. 8, 1959, pages 396
EAGLE, H., SCIENCE, vol. 130, 1959, pages 432
GIL, A. ET AL., CELL, vol. 49, 1987, pages 399406
GIMMI, E. R. ET AL., NUCLEIC ACIDS RES., vol. 17, 1989, pages 6983 - 6998
GOPAL, MOL. CELL BIOL., vol. 5, 1985, pages 1188 - 1190
GRAHAM, F.L. ET AL., VIROLOGY, vol. 52, 1973, pages 456
HAM, PROC. NATL. ACAD SCI. USA, vol. 53, 1965, pages 288
HAM, R.G. ET AL., IN VITRO, vol. 14, 1978, pages 11
HAM, R.G., EXP. CELL RES., vol. 29, 1963, pages 515
ISCOVE, N. ET AL., J. EXP. MED., vol. 147, 1978, pages 923
KAUFINAN ET AL., MOL CELL BIOL. J, vol. 5, no. 7, 1985, pages 1750 - 9
KAUFINAN ET AL., MOL CELL BIOL. JUL, vol. 5, no. 7, 1985, pages 1750 - 9
KLASENS, B. 1. F. ET AL., NUCLEIC ACIDS RES., vol. 26, 1998, pages 1870 - 1876
MCCOY, T.A. ET AL., PROC. SOC. EXP. BIOL. MED., vol. 100, 1959, pages 115
MOORE ET AL., J AMER. MED. ASSOC., vol. 199, 1967, pages 519
MORGAN ET AL., PROC. SOC. EXP. BIO. MED., vol. 73, 1950, pages 1
NEUMANN, E. ET AL., EMBO J, vol. 1, 1982, pages 841
RESEARCH INSTITUTE OF BIOSCIENCE AND BIOTECHNOLOGY, 11 March 2008 (2008-03-11)
SCHEK, N ET AL., MOL. CELL BIOL., vol. 12, 1992, pages 5386 - 5393
See also references of EP2314701A4
STANNER, C.P. ET AL., NAT. NEW BIOL., vol. 230, 1971, pages 52
WAYMOUTH, C., J NATL. CANCER INST., vol. 22, 1959, pages 1003
WONG, T.K. ET AL., GENE, vol. 10, 1980, pages 87
YANG ET AL., PROC. NATL. ACAD SCI., vol. 87, 1990, pages 9568 - 9572

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013544080A (ja) * 2010-10-26 2013-12-12 ハンミ サイエンス カンパニー リミテッド ヒト血液凝固第ｖｉｉ因子の大量生産方法
US9243237B2 (en) 2010-10-26 2016-01-26 Hanmi Science Co., Ltd Method for mass production of factor VII/VIIA

Also Published As

Publication number Publication date
AU2009270108B2 (en) 2013-10-24
EP2314701A4 (en) 2012-03-07
CN102177239A (zh) 2011-09-07
KR101030978B1 (ko) 2011-04-28
WO2010005227A3 (ko) 2010-04-22
CA2733119A1 (en) 2010-01-14
BRPI0915734A2 (pt) 2015-08-04
EP2314701B1 (en) 2015-07-29
CA2733119C (en) 2014-04-08
US20110117643A1 (en) 2011-05-19
CN102177239B (zh) 2014-06-04
JP5514818B2 (ja) 2014-06-04
KR20100006800A (ko) 2010-01-22
WO2010005227A8 (ko) 2010-06-24
EP2314701A2 (en) 2011-04-27
US8426570B2 (en) 2013-04-23
JP2011527567A (ja) 2011-11-04
AU2009270108A1 (en) 2010-01-14

Similar Documents

Publication Publication Date Title
MX2012015142A (es) Polipeptidos que tiene actividad beta-glucosidasa y usos del mismo.
WO2010120514A3 (en) Antigen-binding proteins comprising recombinant protein scaffolds
MY160897A (en) Polypeptide having or assisting in carbohydrate material degrading activity and uses thereof
WO2010043650A3 (en) Amino acid sequences targeting human cd4, cxcr4, ccr5, tlr4, alphav integrin, beta3- integrin, betal -integrin, human alpha2- integrin, cd81, sr-bi, claudin-i, claudin-6 and claudin-9, respectively, and neutralizing viral entry
WO2014118360A3 (en) Carbohydrate degrading polypeptide and uses thereof
MX2013007853A (es) Polipeptidos que tienen actividad de celobiohidrolasa y polinucleotidos que codifican los mismos.
MX2014002497A (es) Sintasa isopropilmalato de nicotiana tabacum y metodo y usos de la misma.
MX337919B (es) Polipeptidos que tienen actividad de celobiohidrolasa y polinucleotidos que codifican los mismos.
WO2007003582A3 (en) Serum-free stable transfection and production of recombinant human proteins in human cell lines
MX337913B (es) Polipeptidos que tienen actividad celobiohidrolasa y polinucleotidos que codifican los mismos.
WO2012074277A3 (en) Novel hybrid promoter and recombinant vector comprising the same
WO2010139736A3 (de) Rekombinante herstellung von peptiden
WO2014097113A3 (en) Production of therapeutic proteins in genetically modified mammalian cells
MY171544A (en) A cell with reduced ppgppase activity
EA201170715A1 (ru) Вектор, содержащий множественные гомологичные нуклеотидные последовательности
PH12015500771A1 (en) Novel fc gamma receptor iib variants
WO2010042755A3 (en) Chimeric therapeutics, compositions, and methods for using same
NZ742660A (en) Variants of chymosin with improved milk-clotting properties
CA2863578C (en) Methods and compositions for increasing sialic acid production and treating sialic related disease conditions
EP2534168A4 (en) CHEMICAL PREPARATION OF UBIQUITIN THIOESTERS AND MODIFICATIONS THEREOF
WO2007130543A3 (en) Method for producing stable mammalian cell lines producing high levels of recombinant proteins
MX2010001491A (es) Proteina precursora de tiopeptidos, gen que la codifica, y usos de la misma.
WO2010005227A2 (ko) 동물세포용 재조합 발현벡터
WO2010127219A3 (en) Altering enzyme balance through fermentation conditions
WO2009125999A8 (en) Human host cell for producing recombinant proteins with high quality and quantity

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 200980126484.1

Country of ref document: CN

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09794633

Country of ref document: EP

Kind code of ref document: A2

WWE Wipo information: entry into national phase

Ref document number: 13002814

Country of ref document: US

ENP Entry into the national phase

Ref document number: 2733119

Country of ref document: CA

Ref document number: 2011517345

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2009270108

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 2009794633

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2009270108

Country of ref document: AU

Date of ref document: 20090707

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: PI0915734

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20110110