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WO2003080660A3 - Method for the preparation of recombinant mammalian heparin-binding protein (hbp) - Google Patents

Method for the preparation of recombinant mammalian heparin-binding protein (hbp) Download PDF

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Publication number
WO2003080660A3
WO2003080660A3 PCT/DK2003/000207 DK0300207W WO03080660A3 WO 2003080660 A3 WO2003080660 A3 WO 2003080660A3 DK 0300207 W DK0300207 W DK 0300207W WO 03080660 A3 WO03080660 A3 WO 03080660A3
Authority
WO
WIPO (PCT)
Prior art keywords
hbp
preparation
binding protein
fusion protein
relates
Prior art date
Application number
PCT/DK2003/000207
Other languages
French (fr)
Other versions
WO2003080660A2 (en
Inventor
Helle Fabricius Woeldike
Original Assignee
Leukotech As
Helle Fabricius Woeldike
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 Leukotech As, Helle Fabricius Woeldike filed Critical Leukotech As
Priority to AU2003226907A priority Critical patent/AU2003226907A1/en
Publication of WO2003080660A2 publication Critical patent/WO2003080660A2/en
Publication of WO2003080660A3 publication Critical patent/WO2003080660A3/en

Links

Classifications

    • 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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4723—Cationic antimicrobial peptides, e.g. defensins
    • 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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • 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
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • 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
    • C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
    • 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)

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Medicinal Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention relates to methods of making heparin-binding protein (HBP) in a recombinant bacterial expression system. In particular, this invention relates to a method for the preparation of an insoluble fusion protein comprising heparin-binding protein (HBP), a proteolytic cleavage- site, and a second polypeptide in recombinant bacterial cells, said fusion protein being accumulated in inclusion bodies in the cytoplasm of bacterium after expression. The invention further relates to methods of separation of the expressed HBP from inclusion bodies comprising purification of HBP from the second polypeptide and optionally refolding of said HBP. The invention further features the DNA constructs comprising different HBP-fusion proteins. Invention also relates to use of the HBP-fusion protein produced in bacteria for the production of pure HBP and use of the HBP purified from the fusion protein for the preparation of a medicament.
PCT/DK2003/000207 2002-03-27 2003-03-26 Method for the preparation of recombinant mammalian heparin-binding protein (hbp) WO2003080660A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003226907A AU2003226907A1 (en) 2002-03-27 2003-03-26 Method for the preparation of recombinant mammalian heparin-binding protein (hbp)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DKPA200200477 2002-03-27
DKPA200200477 2002-03-27

Publications (2)

Publication Number Publication Date
WO2003080660A2 WO2003080660A2 (en) 2003-10-02
WO2003080660A3 true WO2003080660A3 (en) 2003-12-18

Family

ID=28051662

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK2003/000207 WO2003080660A2 (en) 2002-03-27 2003-03-26 Method for the preparation of recombinant mammalian heparin-binding protein (hbp)

Country Status (2)

Country Link
AU (1) AU2003226907A1 (en)
WO (1) WO2003080660A2 (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150056653A1 (en) * 2011-07-29 2015-02-26 Georgia State University Research Foundation, Inc. Protein production method
CN108018294B (en) * 2018-01-02 2020-11-17 昆明理工大学 Panax notoginseng plant hormone binding protein genePnPhBP1And applications
CN117343200B (en) * 2023-12-04 2024-01-30 北京质肽生物医药科技有限公司 Fusion protein containing amyloid core polypeptide, preparation method and application thereof
CN117965667B (en) * 2024-01-23 2024-08-16 陕西普罗安蒂生物科技发展有限公司 Method for preparing polypeptide by cutting fusion protein

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989008666A1 (en) * 1988-03-17 1989-09-21 Novo-Nordisk A/S Heparin-binding proteins, dna cuding for them, processes for producing them as well as therapeutic preparations containing them
US5484885A (en) * 1989-07-05 1996-01-16 Emory University Chemotactic, antibiotic and lipopolysaccharide-binding peptide fragments of CAP37
WO1999000417A1 (en) * 1997-06-25 1999-01-07 Novo Nordisk A/S Production of heparin-binding protein in mammalian cells
WO1999026647A1 (en) * 1997-11-20 1999-06-03 Novo Nordisk A/S Use of heparin-binding protein for the modulation or prophylaxis of apoptosis of mammalian cells
WO2000066627A1 (en) * 1999-04-29 2000-11-09 Leukotech A/S Expression of heparin-binding protein in recombinant mammalian cells

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989008666A1 (en) * 1988-03-17 1989-09-21 Novo-Nordisk A/S Heparin-binding proteins, dna cuding for them, processes for producing them as well as therapeutic preparations containing them
US5484885A (en) * 1989-07-05 1996-01-16 Emory University Chemotactic, antibiotic and lipopolysaccharide-binding peptide fragments of CAP37
WO1999000417A1 (en) * 1997-06-25 1999-01-07 Novo Nordisk A/S Production of heparin-binding protein in mammalian cells
WO1999026647A1 (en) * 1997-11-20 1999-06-03 Novo Nordisk A/S Use of heparin-binding protein for the modulation or prophylaxis of apoptosis of mammalian cells
WO2000066627A1 (en) * 1999-04-29 2000-11-09 Leukotech A/S Expression of heparin-binding protein in recombinant mammalian cells

Also Published As

Publication number Publication date
AU2003226907A1 (en) 2003-10-08
AU2003226907A8 (en) 2003-10-08
WO2003080660A2 (en) 2003-10-02

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