WO1999001548A3 - Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin - Google Patents

Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin Download PDF

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Publication number
WO1999001548A3
WO1999001548A3 PCT/IB1998/001023 IB9801023W WO9901548A3 WO 1999001548 A3 WO1999001548 A3 WO 1999001548A3 IB 9801023 W IB9801023 W IB 9801023W WO 9901548 A3 WO9901548 A3 WO 9901548A3
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WO
WIPO (PCT)
Prior art keywords
insulin
protein
vector
recombinant
heterologous protein
Prior art date
Application number
PCT/IB1998/001023
Other languages
French (fr)
Other versions
WO1999001548A2 (en
Inventor
Spartaco Astolfi-Filho
Lima Beatriz Dolabela De
Josef Ernst Thiemann
Sousa Heloisa Ribeiro Tunes De
Luciano Vilela
Original Assignee
Univ Brasilia
Biobras S A
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 Univ Brasilia, Biobras S A filed Critical Univ Brasilia
Priority to IL13383298A priority Critical patent/IL133832A0/en
Priority to PL98348669A priority patent/PL194141B1/en
Priority to EP98928476A priority patent/EP0996713B1/en
Priority to CA2294760A priority patent/CA2294760C/en
Priority to DE69826864T priority patent/DE69826864T2/en
Priority to AT98928476T priority patent/ATE278776T1/en
Priority to BRPI9810650-3A priority patent/BR9810650B1/en
Publication of WO1999001548A2 publication Critical patent/WO1999001548A2/en
Publication of WO1999001548A3 publication Critical patent/WO1999001548A3/en
Priority to IL133832A priority patent/IL133832A/en

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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
    • 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
    • C12N15/70Vectors or expression systems specially adapted for E. coli
    • C12N15/73Expression systems using phage (lambda) regulatory sequences
    • 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
    • C07K14/575Hormones
    • C07K14/62Insulins
    • 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
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • 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
    • C12N15/64General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00Preparation of peptides or proteins
    • C12P21/02Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Chemical & Material Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Medicinal Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Endocrinology (AREA)
  • Diabetes (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Cell Biology (AREA)
  • Virology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The present invention relates to a vector for expression of a heterologous protein by a Gram negative bacteria, wherein the vector includes a nucleic acid such as DNA encoding the following: an origin of replication region; optionally and preferably a selection marker; a promoter; an initiation region such as translation initiation region and/or a ribosome binding site, at least one restriction site for insertion of heterologous nucleic acid, e.g. DNA, encoding the heterologous protein, and a transcription terminator. The inventive vector may contain DNA encoding the heterologous protein, e.g., pro-insulin such as pro-insulin with a His tag. Additionally, the invention provides a method for extracting a recombinant protein from within a recombinant Gram negative bacteria having a cell membrane, without lysing the bacteria, as well as a method for purifying an isolated recombinant human insulin, wherein the isolated recombinant human pro-insulin is subjected to sulfitolysis, Ni-chelation chromatography, renaturation, limited proteolysis and chromatography separation to provide purified, isolated, recombinant human insulin.
PCT/IB1998/001023 1997-07-02 1998-07-02 Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin WO1999001548A2 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
IL13383298A IL133832A0 (en) 1997-07-02 1998-07-02 Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin
PL98348669A PL194141B1 (en) 1997-07-02 1998-07-02 Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin
EP98928476A EP0996713B1 (en) 1997-07-02 1998-07-02 Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin
CA2294760A CA2294760C (en) 1997-07-02 1998-07-02 Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin
DE69826864T DE69826864T2 (en) 1997-07-02 1998-07-02 VECTOR FOR HETEROLOGICAL PROTEIN EXPRESSION AND METHOD FOR EXTRACTION OF RECOMBINANT PROTEINS AND FOR INSULATION OF RECOMBINANT INSULINS
AT98928476T ATE278776T1 (en) 1997-07-02 1998-07-02 VECTOR FOR HETEROLOGUE PROTEIN EXPRESSION AND METHOD FOR EXTRACTION OF RECOMBINANT PROTEINS AND FOR ISOLATION OF RECOMBINANT INSULINS
BRPI9810650-3A BR9810650B1 (en) 1997-07-02 1998-07-02 vector for heterologous protein expression and methods for extracting recombinant protein and for purifying isolated recombinant insulin.
IL133832A IL133832A (en) 1997-07-02 1999-12-30 Expression vector for transforming gram negative bacteria and process for producing a heterologous protein from the vector

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/886,967 US6068993A (en) 1997-07-02 1997-07-02 Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin
US08/886,967 1997-07-02

Publications (2)

Publication Number Publication Date
WO1999001548A2 WO1999001548A2 (en) 1999-01-14
WO1999001548A3 true WO1999001548A3 (en) 1999-09-10

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB1998/001023 WO1999001548A2 (en) 1997-07-02 1998-07-02 Vector for expression of heterologous protein and methods for extracting recombinant protein and for purifying isolated recombinant insulin

Country Status (11)

Country Link
US (4) US6068993A (en)
EP (1) EP0996713B1 (en)
AT (1) ATE278776T1 (en)
BR (1) BR9810650B1 (en)
CA (2) CA2687542C (en)
DE (1) DE69826864T2 (en)
IL (2) IL133832A0 (en)
IN (1) IN187721B (en)
PL (1) PL194141B1 (en)
RU (1) RU2222598C2 (en)
WO (1) WO1999001548A2 (en)

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US7049286B2 (en) * 2001-08-30 2006-05-23 Diatos, S.A. Insulin conjugates and methods of use thereof
WO2004042003A2 (en) * 2002-11-01 2004-05-21 Promega Corporation Cell lysis compositions, methods of use, apparatus, and kit
EP1670446A2 (en) 2003-09-26 2006-06-21 Japan Tobacco Inc. Method of inhibiting remnant lipoprotein production
US7674621B2 (en) 2004-05-21 2010-03-09 The United States Of America As Represented By The Department Of Health And Human Services Plasmids and phages for homologous recombination and methods of use
CN105801686B (en) 2004-07-19 2020-04-07 比奥孔有限公司 Insulin-oligomer conjugates, formulations and uses thereof
WO2006037215A1 (en) * 2004-10-04 2006-04-13 National Research Council Of Canada Expression system, components thereof and methods of use
US7935788B2 (en) * 2004-10-04 2011-05-03 National Research Council Of Canada Reverse cumate repressor mutant
WO2009032664A2 (en) * 2007-08-31 2009-03-12 The Regents Of The University Of Michigan Methods and compositions for increasing biological molecule stability
EP2203181B1 (en) 2007-10-16 2018-02-14 Biocon Limited An orally administerable solid pharmaceutical composition and a process thereof
ES2799431T3 (en) * 2010-03-31 2020-12-17 Amsilk Gmbh Insoluble target protein separation
RU2497118C1 (en) * 2012-03-06 2013-10-27 Федеральное государственное бюджетное учреждение науки Институт биохимии и физиологии микроорганизмов им. Г.К. Скрябина Российской академии наук (ИБФМ РАН) FAST METHOD FOR TREATING INTESTINAL SMOOTH MUSCLE CELLS WITH Bacillus cereus HEMOLYSIN II
CN102617703A (en) * 2012-04-14 2012-08-01 拜明(苏州)生物技术有限公司 Method for extracting exogenous expressive protein in fermentation bacteria efficiently without cracking
WO2015176067A2 (en) * 2014-05-16 2015-11-19 The Regents Of The University Of California A long non-coding rna expressed in aggressive cancer
EP3344651B1 (en) 2015-09-02 2022-03-02 Merck Sharp & Dohme Corp. A process for obtaining insulin with correctly formed disulfide bonds

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Also Published As

Publication number Publication date
BR9810650A (en) 2002-07-23
WO1999001548A2 (en) 1999-01-14
EP0996713A1 (en) 2000-05-03
DE69826864D1 (en) 2004-11-11
US6699692B2 (en) 2004-03-02
PL194141B1 (en) 2007-04-30
CA2294760C (en) 2010-02-23
CA2687542C (en) 2013-04-30
PL348669A1 (en) 2002-06-03
US6281329B1 (en) 2001-08-28
DE69826864T2 (en) 2005-11-03
US6068993A (en) 2000-05-30
EP0996713B1 (en) 2004-10-06
CA2687542A1 (en) 1999-01-14
BR9810650B1 (en) 2010-12-14
US6509452B1 (en) 2003-01-21
IL133832A0 (en) 2001-04-30
IL133832A (en) 2007-07-04
US20030082742A1 (en) 2003-05-01
IN187721B (en) 2002-06-15
ATE278776T1 (en) 2004-10-15
RU2222598C2 (en) 2004-01-27
CA2294760A1 (en) 1999-01-14

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