FI105482B - Förfarande för framställning av ett heterologt protein i en eukaryotisk värdcell - Google Patents

Förfarande för framställning av ett heterologt protein i en eukaryotisk värdcell Download PDF

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Publication number
FI105482B
FI105482B FI925678A FI925678A FI105482B FI 105482 B FI105482 B FI 105482B FI 925678 A FI925678 A FI 925678A FI 925678 A FI925678 A FI 925678A FI 105482 B FI105482 B FI 105482B
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Finland
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sequence
kex2
yeast
host cell
protein
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FI925678A
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English (en)
Finnish (fi)
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FI925678A0 (fi
FI925678A (fi
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Bhabatosh Chaudhuri
Howard Riezman
Christine Stephan
Peter Seeboth
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Novartis Ag
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Publication of FI105482B publication Critical patent/FI105482B/sv

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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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
    • C07K1/1133—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure by redox-reactions involving cystein/cystin side chains
    • 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
    • C07K14/575—Hormones
    • C07K14/65—Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • 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
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/58—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi
    • C12N9/60—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi from yeast
    • 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
    • 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
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/04—Fusion polypeptide containing a localisation/targetting motif containing an ER retention signal such as a C-terminal HDEL motif
    • 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
    • C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones

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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)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Mycology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Endocrinology (AREA)
  • Diabetes (AREA)
  • Analytical Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Saccharide Compounds (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Claims (13)

1. Förfarande för framstälining av ett heterologt protein i en eukaryotisk värdcell, kännetecknat därav, att en 5 värdcell transformeras med en nukleinsyra, vilken kodar för bäde en pro-sekvens innehällande pro-form av ett protein och ett tväbasiskt, bearbetande endoproteas, vil-ket förmär avlägsna pro-sekvensen och är fusionerat tili en retentionssignal av endoplasmatisk reticulum; och värd-10 cellen odlas för att producera det heterologa proteinet.
2. Förfarande enligt patentkravet 1, kännetecknat därav, att skilda nukleinsyror kodar för ett heterologt protein och ett tväbasiskt, bearbetande endoproteas. 15
3. Förfarande enligt patentkravet 2, kännetecknat därav, att värdcellen permanent transformeras med en nukleinsyra, vilken kodar för ett tväbasiskt, bearbetande endoproteas och därefter ännu transformeras med en nukleinsyra, vilken 20 kodar för ett heterologt protein.
4. Förfarande enligt nägot av patentkraven 1-3, känne- • « « tecknat därav, att värdcellen är en jästcell och det : *" tväbasiska, bearbetande endoproteaset är KEX2 eller YAP3. 25
• · ·.{ : 5. Förfarande enligt nägot av de föregäende patentkraven, kännetecknat därav, att pro-sekvensen är en pro-sekvens av • · jäst a-faktor. .···. 30
6. Förfarande enligt nägot av de föregäende patentkraven, kännetecknat därav, att det heterologa proteinet är ett *1* biologiskt aktivt protein. • · • i » I f ' 4. i. .'ti
: 7. Förfarande enligt patentkravet 6, kännetecknat därav, . 35 att det biologiskt aktiva proteinet är ett insulin liknan- de protein. Hill * 105482
8. Rekombinant DNA-molekyl, vilken innehäller en expres-sionskasett, vilken kodar för ett tväbasiskt, bearbetande endoproteas, vilket Sr fusionerat till en retentionssignal av endoplasmatisk reticulum. 5
9. Rekombinant DNA-molekyl enligt patentkravet 8, känne-tecknad därav, att det tväbasiska, bearbetande endopro-teaset är lösligt.
10. Rekombinant DNA-molekyl enligt patentkravet 8, känne tecknad därav, att det tväbasiska, bearbetande endopro-teaset Sr KEX2 med en C-terminal extension HDEL, lösligt KEX2 vars aminosyrasekvens Sr visad i sekvensen nr. l och med en C-terminal extension HDEL, eller YAP3 med en C-15 terminal extension HDEL.
11. Hybridvektor, kännetecknad därav, att den innefattar en rekombinant DNA-molekyl enligt nSgot av patentkraven 8- 10. 20
12. Eukaryotisk värdcell, vilken Sr transformerad med en vektor enligt patentkravet 11.
• · · : 13. Eukaryotisk värdcell, kännetecknad därav, att den Sr !,·,·' 25 stabilt transformerad. I « • > I • · · • · · · • · • · · • · · • · • · · • · · • · · • · · t 1 « · ·· r I I f · • « • · • · · • f · « t ·
FI925678A 1991-12-16 1992-12-14 Förfarande för framställning av ett heterologt protein i en eukaryotisk värdcell FI105482B (sv)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP91810984 1991-12-16
EP91810984 1991-12-16

Publications (3)

Publication Number Publication Date
FI925678A0 FI925678A0 (fi) 1992-12-14
FI925678A FI925678A (fi) 1993-06-17
FI105482B true FI105482B (sv) 2000-08-31

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Family Applications (1)

Application Number Title Priority Date Filing Date
FI925678A FI105482B (sv) 1991-12-16 1992-12-14 Förfarande för framställning av ett heterologt protein i en eukaryotisk värdcell

Country Status (17)

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US (2) US5501975A (sv)
EP (1) EP0548012B1 (sv)
JP (1) JP3305781B2 (sv)
KR (1) KR100270283B1 (sv)
AT (1) ATE157703T1 (sv)
AU (1) AU667852B2 (sv)
CA (1) CA2085308C (sv)
DE (1) DE69222013T2 (sv)
DK (1) DK0548012T3 (sv)
ES (1) ES2107520T3 (sv)
FI (1) FI105482B (sv)
GR (1) GR3025324T3 (sv)
IL (1) IL104075A (sv)
MX (1) MX9207291A (sv)
NO (1) NO308619B1 (sv)
NZ (1) NZ245469A (sv)
ZA (1) ZA929711B (sv)

Families Citing this family (16)

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Publication number Priority date Publication date Assignee Title
GB9404270D0 (en) * 1994-03-05 1994-04-20 Delta Biotechnology Ltd Yeast strains and modified albumins
US5807702A (en) * 1995-02-27 1998-09-15 Abbott Laboratories Method for expressing phosphorylated recombinant human β-casein in a bacterial system
US5710044A (en) * 1995-02-27 1998-01-20 Abbott Laboratories Plasmid for expressing phosphorylated recombinant proteins in a bacterial system
EP0812362A1 (en) * 1995-02-27 1997-12-17 Abbott Laboratories A plasmid for expressing modified recombinant proteins in a bacterial system
NZ304538A (en) * 1995-02-27 1999-08-30 Abbott Lab Process for producing a modified recombinant protein in a host cell comprising preparing a single vector encoding both the exogenous protein and an enzyme capable of modifying the exogenous protein
US5942254A (en) * 1995-02-27 1999-08-24 Abbott Laboratories Phosphorylated recombinant human β-casein expressed in a bacterial system
AT404838B (de) * 1995-11-24 1999-03-25 Immuno Ag Herstellung von proteinen aus pro-proteinen durch fusionsproteine abgeleitet von furin oder furinanalogen
FR2751000B1 (fr) 1996-07-12 1998-10-30 Inst Nat Sante Rech Med Adn specifiques des bacteries de l'espece neisseria meningitidis, leurs procedes d'obtention et leurs applications biologiques
DE69939548D1 (de) * 1998-11-06 2008-10-23 Novo Nordisk Healthcare Ag Verfahren zur herstellung von faktor vii
AU7765801A (en) * 2000-06-30 2002-01-08 Flanders Interuniversity Inst Protein glycosylation modification in pichia pastoris
DE10221411B4 (de) * 2002-05-14 2004-07-08 GSF-Forschungszentrum für Umwelt und Gesundheit GmbH Rekombinantes Fowlpox-Virus
US20040117874A1 (en) * 2002-10-15 2004-06-17 Jianjun Yang Methods for accumulating translocated proteins
US7507573B2 (en) 2003-11-14 2009-03-24 Vib, Vzw Modification of protein glycosylation in methylotrophic yeast
US7459548B2 (en) * 2004-05-21 2008-12-02 Mo Bio Laboratories, Inc. Kits and processes for removing contaminants from nucleic acids in environmental and biological samples
US20080082338A1 (en) * 2006-09-29 2008-04-03 O'neil Michael P Systems and methods for secure voice identification and medical device interface
WO2014138371A1 (en) 2013-03-06 2014-09-12 Glaxosmithkline Llc Host cells and methods of use

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3685996T2 (de) * 1985-06-11 1993-01-14 Ciba Geigy Ag Hybrid-interferone.
US5391485A (en) * 1985-08-06 1995-02-21 Immunex Corporation DNAs encoding analog GM-CSF molecules displaying resistance to proteases which cleave at adjacent dibasic residues
FI100106B (sv) * 1986-12-05 1997-09-30 Novartis Ag Förfarande för framställning av en enkelsträngad hybridplasminogenakti vator
JP2643968B2 (ja) * 1988-02-03 1997-08-25 サントリー株式会社 Ｋｅｘ２エンドプロテアーゼ及びその製造方法

Also Published As

Publication number Publication date
ZA929711B (en) 1993-06-16
GR3025324T3 (en) 1998-02-27
NZ245469A (en) 1995-03-28
AU667852B2 (en) 1996-04-18
EP0548012B1 (en) 1997-09-03
DE69222013T2 (de) 1998-01-29
US5618690A (en) 1997-04-08
CA2085308C (en) 2003-12-02
AU3007192A (en) 1993-06-17
JPH06197787A (ja) 1994-07-19
US5501975A (en) 1996-03-26
IL104075A0 (en) 1993-05-13
KR930013115A (ko) 1993-07-21
ES2107520T3 (es) 1997-12-01
EP0548012A1 (en) 1993-06-23
CA2085308A1 (en) 1993-06-17
NO308619B1 (no) 2000-10-02
FI925678A0 (fi) 1992-12-14
FI925678A (fi) 1993-06-17
KR100270283B1 (ko) 2000-11-01
ATE157703T1 (de) 1997-09-15
JP3305781B2 (ja) 2002-07-24
DE69222013D1 (de) 1997-10-09
DK0548012T3 (da) 1998-04-14
MX9207291A (es) 1993-07-01
NO924847D0 (no) 1992-12-15
NO924847L (no) 1993-06-17
IL104075A (en) 2003-01-12

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