FI113372B - Förfarande för framställning av en LIF-polypeptid (LIF = leukemiinhiberande faktor) - Google Patents

Förfarande för framställning av en LIF-polypeptid (LIF = leukemiinhiberande faktor) Download PDF

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FI113372B
FI113372B FI894613A FI894613A FI113372B FI 113372 B FI113372 B FI 113372B FI 894613 A FI894613 A FI 894613A FI 894613 A FI894613 A FI 894613A FI 113372 B FI113372 B FI 113372B
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lif
cells
mouse
polypeptide
human
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FI894613A
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Finnish (fi)
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FI894613A (fi
FI894613A0 (fi
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Donald Metcalf
Nicholas Martin Gough
Douglas James Hilton
David Paul Gearing
Julie Ann King
Edouard Collins Nice
Nicos Anthony Nicola
Richard John Simpson
Tracy Ann Willson
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Amrad Corp Ltd
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    • C—CHEMISTRY; METALLURGY
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    • 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
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/54—Interleukins [IL]
    • C07K14/5415—Leukaemia inhibitory factor [LIF]
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
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    • 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
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • 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
    • 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
    • C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/23—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a GST-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/50—Fusion polypeptide containing protease site
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/95—Fusion polypeptide containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence)
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/13011—Gammaretrovirus, e.g. murine leukeamia virus
    • C12N2740/13041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/13043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
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  • Genetics & Genomics (AREA)
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  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
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  • Mycology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Gastroenterology & Hepatology (AREA)
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  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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  • Tropical Medicine & Parasitology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Claims (21)

1. Förfarande för framställning av en LIF-polypeptid (LIF = leukemiinhiberande faktor), kännetecknat därav, att det omfattar: (a) framställning av värdceller, som innehäller en rekombinant-DNA-molekyl, som kodar för LIF-polypeptiden, i kombination med transkriptions- öch translationsreglerande sekvenser samt selektionsmarkörer, (b) odling av dessa celler under betingelser, som leder tili en expression av polypeptiden, och (c) utvinning av polypeptiden sä, att polypeptiden omfattar en mogen polypeptid, som tili minst 78 % är homolog med en aminosyrasekvens utvald ur aminosyrasekvenserna visade i figurerna 15, 26 och 29.
2. Förfarande enligt patentkrav 1, kännetecknat därav, att den mogna polypeptiden uppvisar fullständig homologi med den i figur 29 visade aminosyrasekvensen.
3. Förfarande enligt patentkrav 1, kännetecknat därav, att polypeptiden är glykosylerad med hjälp av värdcellen. • * *» #
4. Förfarande enligt patentkrav 1, , k ä n net e c k n a t därav, att reglerande sekvenserna : : omfattar en promoter som är en galaktosinducerbar hybrid-GAL- CYC-promotor.
5. Förfarande enligt patentkrav 1, kännetecknat därav, att signalsekvensen, som kodär för expressionen av en reglersekvens som styr sekretionen av polypeptiden, som avletts ur signalsekvensen i : killertoxingenen frän Kluyveromyces latis.
6. Förfarande enligt patentkrav 1, kännetecknat därav, att nämnda DNA-molekyl är en vektor, som har avletts frän ett Moloney murint leukemivirus. 67 1 13 3 7 2
7. Förfarande enligt patentkrav 6, kännetecknat därav, att vektorn ytterligare omfattar LTR-enhancern av myeloproliferativt sarkomavirus.
8. Förfarande enligt patentkrav 6, kännetecknat därav, att den nämnda vektorn saknar det 31icke-translaterade omrädet i nativ LIF-mRNA.
9. Förfarande enligt patentkrav 1, kännetecknat därav, att vektorn är en sadan som styr expressionen av ett glutation-s-transferas/LIF-fusionsprotein.
10. Förfarande enligt patentkrav 9, kännetecknat därav, att vektorn är en sadan som styr expressionen av ett glutation-S-transferas/LIF-fusionsprotein/trombinspaltningsstäile/LIF-fusionsprotein.
11. Rekombinant DNA-molekyl, som kodar för en LIF-polypeptid, kännetecknad därav, att den omfattar en i figurerna 10, 15 (baserna 1-63), 25 eller 29 (baserna 1-543) beskriven nukleotidsekvensen, när den nämnda polypeptiden konkurrerar med en polypeptid med en aminosyrasekvens beskriven i figurerna 10, 15, 26 eller 29 för bindning tili specifika • · ‘t cellulära receptorer pä Ml-celler eller murina eller hiomana * ' * * makrofager.
12. Rekdmbinant DNA-molekyl enligt patentkrav 11 omfattande en nukleotidsekvens, sora vid expression i en lämplig värdcell kodar : för en LIF-peptid, kännetecknad därav, att den nämnda nukleotidsekvensen hydridiseras under betingelser med 6xSSC, . ·, 65 C eller härdare betingelser, tili en sekvens av ett LIF- kodande inskott valt frän de LIF-kodande inskotten i klonerna ' pLIF7.2b, pLIFNKl, pLIFNK3 och pHGLIFBaml som derinierats i figurerna 10, 11, 15 och 25.
13. Molekyl enligt patentkrav 12, kännetecknad därav, att den färdiga polypeptiden uppvisar fullständig homologi med aminosyrasekvensen visad i 68 1 13 3 7 2 figur 29.
14. Molekyl enligt patentkrav 11-13, kännetecknad därav, att den ytterligare omfattar en replikationsutgängspunkt, varvid det nämnda molekylen görs lämplig för användning som kloningsvektor.
15. Molekyl enligt patentkrav 14, kännetecknad därav, att den ytterligare omfattar en promotorsekvens som är operabelt bunden tili den nämnda nukleotidsekvensen, varvid den nämnda molekylen görs lämplig för användning som expressionsvektor.
16. Värdcell, kännetecknad därav, att den är transformerad med en rekombinant DNA-molekyl enligt patentkrav 11 och att den förmlr gestalta LIF-peptiden..
17. Värdcell enligt patentkrav 16, kännetecknaddärav, att värdcellen är en jästcell.
18. värdcell enligt patentkrav 17, > kännetecknad därav, att värdcellen är av arten . Saccharomyces cerevisiae. » * · ' t
19. Värdcell enligt patentkrav 16, » * k anne tee n ad därav, att värdcellen är en däggdjurcell. .
20. Värdcell enligt patentkrav 19, : kännetecknad därav, att värdcellerna är blodbildande celler. I I • I» ♦ iti'*
‘ 21. Värdcell enligt. patentkrav 16, kanne, tecknad därav, att värdcellerna är ί E. coli -celler.
FI894613A 1987-04-02 1989-09-29 Förfarande för framställning av en LIF-polypeptid (LIF = leukemiinhiberande faktor) FI113372B (sv)

Applications Claiming Priority (10)

Application Number Priority Date Filing Date Title
AUPI120987 1987-04-02
AUPI120987 1987-04-02
AUPI331787 1987-07-24
AUPI331787 1987-07-24
AUPI490387 1987-10-15
AUPI490387 1987-10-15
AUPI600587 1987-12-21
AUPI600587 1987-12-21
AU8800093 1988-03-31
PCT/AU1988/000093 WO1988007548A1 (en) 1987-04-02 1988-03-31 Leukaemia-inhibitory factor

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FI894613A FI894613A (fi) 1989-09-29
FI894613A0 FI894613A0 (fi) 1989-09-29
FI113372B true FI113372B (sv) 2004-04-15

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FI894613A FI113372B (sv) 1987-04-02 1989-09-29 Förfarande för framställning av en LIF-polypeptid (LIF = leukemiinhiberande faktor)

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US (5) US5187077A (sv)
EP (1) EP0285448B1 (sv)
JP (2) JP2682858B2 (sv)
KR (1) KR0121324B1 (sv)
CA (1) CA1341581C (sv)
DE (1) DE3888379T2 (sv)
DK (1) DK174970B1 (sv)
ES (1) ES2061643T3 (sv)
FI (1) FI113372B (sv)
HK (1) HK33695A (sv)
IL (1) IL85961A (sv)
NO (1) NO178265C (sv)
PT (1) PT87133B (sv)
SG (1) SG11795G (sv)
WO (1) WO1988007548A1 (sv)

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JPWO2011125948A1 (ja) 2010-04-02 2013-07-11 独立行政法人理化学研究所 Ｅｓ細胞の製造方法
KR101434739B1 (ko) * 2012-08-23 2014-09-01 울산대학교 산학협력단 인간 ＰＤＩｂ´ａ´태그를 이용한 생물학적 활성을 가진 인간 ＬＩＦ 재조합 단백질의 대장균 내 발현 및 정제방법
CN109913495B (zh) 2013-02-20 2022-11-25 瑞泽恩制药公司 大鼠的遗传修饰
CN111500630A (zh) 2013-04-16 2020-08-07 瑞泽恩制药公司 大鼠基因组的靶向修饰
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PT87133B (pt) * 1987-04-02 1992-07-31 Amrad Corp Ltd Metodo de purificacao do factor inibidor da leucemia (lif) e de composicoes farmaceuticas contendo polipeptidos com actividade do lif
JPS6448197A (en) * 1987-08-19 1989-02-22 Sanden Corp Goods takeoff port unit for vending machine
JPH01148197A (ja) * 1987-12-04 1989-06-09 Green Cross Corp:The 腫瘍細胞障害因子

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EP0285448A2 (en) 1988-10-05
HK33695A (en) 1995-03-17
DE3888379T2 (de) 1994-07-28
NO885339D0 (no) 1988-11-30
NO885339L (no) 1989-02-01
JP2721123B2 (ja) 1998-03-04
DK483189D0 (da) 1989-10-02
ES2061643T3 (es) 1994-12-16
US5443825A (en) 1995-08-22
JPH01502985A (ja) 1989-10-12
EP0285448B1 (en) 1994-03-16
US5187077A (en) 1993-02-16
DE3888379D1 (de) 1994-04-21
US5750654A (en) 1998-05-12
NO178265C (no) 1996-02-21
JP2682858B2 (ja) 1997-11-26
SG11795G (en) 1995-06-16
CA1341581C (en) 2008-09-30
KR0121324B1 (ko) 1997-11-28
DK483189A (da) 1989-12-04
FI894613A (fi) 1989-09-29
US5427925A (en) 1995-06-27
WO1988007548A1 (en) 1988-10-06
FI894613A0 (fi) 1989-09-29
IL85961A (en) 1997-03-18
NO178265B (no) 1995-11-13
PT87133A (pt) 1988-04-01
US6261548B1 (en) 2001-07-17
JPH07138292A (ja) 1995-05-30
DK174970B1 (da) 2004-04-05
EP0285448A3 (en) 1990-08-29
PT87133B (pt) 1992-07-31
KR890700610A (ko) 1989-04-26

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