WO2003025116A3 - Verfahren zur erhöhung der löslichkeit, der expressionsrate und der aktivität von proteinen während der rekombinanten herstellung - Google Patents

Verfahren zur erhöhung der löslichkeit, der expressionsrate und der aktivität von proteinen während der rekombinanten herstellung Download PDF

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Publication number
WO2003025116A3
WO2003025116A3 PCT/EP2002/010329 EP0210329W WO03025116A3 WO 2003025116 A3 WO2003025116 A3 WO 2003025116A3 EP 0210329 W EP0210329 W EP 0210329W WO 03025116 A3 WO03025116 A3 WO 03025116A3
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WO
WIPO (PCT)
Prior art keywords
solubility
activity
increasing
recombinant production
expression rate
Prior art date
Application number
PCT/EP2002/010329
Other languages
English (en)
French (fr)
Other versions
WO2003025116A2 (de
Inventor
Manfred Watzele
Regina Schweizer
Cordula Nemetz
Robin Steigerwald
Thomas Emrich
Katrin Zaiss
Erhard Fernholz
Baerbel Walckhoff
Hans Joachim Schoenfeld
Birgit Offen
Original Assignee
Roche Diagnostics Gmbh
Hoffmann La Roche
Manfred Watzele
Regina Schweizer
Cordula Nemetz
Robin Steigerwald
Thomas Emrich
Katrin Zaiss
Erhard Fernholz
Baerbel Walckhoff
Hans Joachim Schoenfeld
Birgit Offen
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 Roche Diagnostics Gmbh, Hoffmann La Roche, Manfred Watzele, Regina Schweizer, Cordula Nemetz, Robin Steigerwald, Thomas Emrich, Katrin Zaiss, Erhard Fernholz, Baerbel Walckhoff, Hans Joachim Schoenfeld, Birgit Offen filed Critical Roche Diagnostics Gmbh
Priority to EP02774617A priority Critical patent/EP1430078A2/de
Priority to JP2003529890A priority patent/JP2005503157A/ja
Priority to US10/489,941 priority patent/US20040248238A1/en
Publication of WO2003025116A2 publication Critical patent/WO2003025116A2/de
Publication of WO2003025116A3 publication Critical patent/WO2003025116A3/de

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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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • 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
    • C12N15/67—General methods for enhancing the expression

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

Abstract

Gegenstand der vorliegenden Erfindung ist ein Verfahren zur Herstellung eines Lysates enthaltend Helferproteine, wobei ein Stamm, welcher geeignet ist für die Gewinnung von in vitro Translations-Lysaten, transformiert wird mit einem Vektor enthaltend ein oder mehrere für ein oder mehrere Helferproteine kodierende Gene, wobei die Helferproteine in diesem Stamm ex-primiert werden und wobei das Lysat enthaltend Helferproteine aus diesen Stämmen gewonnen wird. Des weiteren ist Gegenstand der vorliegenden Erfindung ein Lysat enthaltend Helferproteine erhältlich nach dem erfindungsgemässen Verfahren, Verschnitte aus diesen Lysaten und die Verwendung der Lysat und Verschnitte in in vitro Translationssystemen.
PCT/EP2002/010329 2001-09-17 2002-09-14 Verfahren zur erhöhung der löslichkeit, der expressionsrate und der aktivität von proteinen während der rekombinanten herstellung WO2003025116A2 (de)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP02774617A EP1430078A2 (de) 2001-09-17 2002-09-14 Verfahren zur erhöhung der löslichkeit, der expressionsrate und der aktivität von proteinen während der rekombinanten herstellung
JP2003529890A JP2005503157A (ja) 2001-09-17 2002-09-14 組換え生産においてタンパク質の溶解性、発現率及び活性を増大させるための方法
US10/489,941 US20040248238A1 (en) 2001-09-17 2002-09-14 Method for increasing the solubility, expression rate and the acitivity of proteins during recombinant production

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10145694.8 2001-09-17
DE10145694A DE10145694A1 (de) 2001-09-17 2001-09-17 Verfahren zur Erhöhung der Löslichkeit, der Expressionsrate und der Aktivität von Proteinen während der rekombinanten Herstellung

Publications (2)

Publication Number Publication Date
WO2003025116A2 WO2003025116A2 (de) 2003-03-27
WO2003025116A3 true WO2003025116A3 (de) 2004-03-11

Family

ID=7699261

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2002/010329 WO2003025116A2 (de) 2001-09-17 2002-09-14 Verfahren zur erhöhung der löslichkeit, der expressionsrate und der aktivität von proteinen während der rekombinanten herstellung

Country Status (5)

Country Link
US (1) US20040248238A1 (de)
EP (1) EP1430078A2 (de)
JP (1) JP2005503157A (de)
DE (1) DE10145694A1 (de)
WO (1) WO2003025116A2 (de)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB0329681D0 (en) * 2003-12-23 2004-01-28 Delta Biotechnology Ltd Gene expression technique
US9057061B2 (en) 2003-12-23 2015-06-16 Novozymes Biopharma Dk A/S Gene expression technique
GB0512707D0 (en) * 2005-06-22 2005-07-27 Delta Biotechnology Ltd Gene expression technique
WO2014172631A2 (en) * 2013-04-19 2014-10-23 Sutro Biopharma, Inc. Expression of biologically active proteins in a bacterial cell-free synthesis system using bacterial cells transformed to exhibit elevated levels of chaperone expression

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0885967A2 (de) * 1997-06-20 1998-12-23 Hsp Research Institute, Inc. Expressionsplasmide für Chaperon (Molekulärkappe)
EP1077262A1 (de) * 1999-07-29 2001-02-21 F. Hoffmann-La Roche Ag Verfahren zur Herstellung von natürlich gefalteten Proteinen durch Co-Sekretion von molekularen Chaperonen

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0698790A (ja) * 1992-03-04 1994-04-12 Kobe Steel Ltd 無細胞蛋白質合成系によるポリペプチド の製造法
EP0693131A4 (de) * 1993-04-08 1998-06-10 Res Dev Foundation Zellfreies system für die proteinsynthese und die verwendung von chaperonproteinen dabei
JP3255784B2 (ja) * 1994-01-26 2002-02-12 弥重太 遠藤 蛋白質の合成方法
DE10121235A1 (de) * 2001-04-30 2002-10-31 Roche Diagnostics Gmbh Verfahren zur Expression von Proteinen in in vitro Translations-Systemen unter Ko-Expression von Faltungshelferproteinen

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0885967A2 (de) * 1997-06-20 1998-12-23 Hsp Research Institute, Inc. Expressionsplasmide für Chaperon (Molekulärkappe)
EP1077262A1 (de) * 1999-07-29 2001-02-21 F. Hoffmann-La Roche Ag Verfahren zur Herstellung von natürlich gefalteten Proteinen durch Co-Sekretion von molekularen Chaperonen
EP1077263A1 (de) * 1999-07-29 2001-02-21 F.Hoffmann-La Roche Ag Verfahren zur Herstellung von natürlich gefalteten und sekretierten Proteinen durch Co-Sekretion von Chaperonen

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
FORSYTHE HEIDI L ET AL: "Stable association of hsp90 and p23, but not hsp70, with active human telomerase.", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 276, no. 19, 11 May 2001 (2001-05-11), pages 15571 - 15574, XP002248678, ISSN: 0021-9258 *

Also Published As

Publication number Publication date
US20040248238A1 (en) 2004-12-09
JP2005503157A (ja) 2005-02-03
DE10145694A1 (de) 2003-04-03
WO2003025116A2 (de) 2003-03-27
EP1430078A2 (de) 2004-06-23

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