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WO1986000339A1 - Procede et appareil de recuperation d'un polypeptide dans un bouillon de fermentation - Google Patents

Procede et appareil de recuperation d'un polypeptide dans un bouillon de fermentation Download PDF

Info

Publication number
WO1986000339A1
WO1986000339A1 PCT/DK1985/000062 DK8500062W WO8600339A1 WO 1986000339 A1 WO1986000339 A1 WO 1986000339A1 DK 8500062 W DK8500062 W DK 8500062W WO 8600339 A1 WO8600339 A1 WO 8600339A1
Authority
WO
WIPO (PCT)
Prior art keywords
fermentation broth
fermentation
polypeptide
process according
microorganism
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/DK1985/000062
Other languages
English (en)
Inventor
John Pedersen
Kim Ry HEJNA^ES
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nordisk Gentofte AS
Original Assignee
Nordisk Gentofte AS
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 Nordisk Gentofte AS filed Critical Nordisk Gentofte AS
Publication of WO1986000339A1 publication Critical patent/WO1986000339A1/fr
Priority to DK84386A priority Critical patent/DK84386D0/da
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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
    • C07K14/575—Hormones
    • C07K14/61—Growth hormone [GH], i.e. somatotropin
    • 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
    • C07K1/14—Extraction; Separation; Purification
    • 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
    • C07K14/62—Insulins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/10—Separation or concentration of fermentation products
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/12—Purification
    • 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
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/582—Recycling of unreacted starting or intermediate materials

Definitions

  • a process and an apparatus for the recovery of a poly ⁇ peptide from a fermentation broth A process and an apparatus for the recovery of a poly ⁇ peptide from a fermentation broth.
  • the present invention concerns a process for recovering a polypeptide from a fermentation broth.
  • Polypeptides which were previously recovered from ani ⁇ mal tissue or organs, e.g. by extraction, are now increas ⁇ ingly produced by bio-technjcal processes using micro ⁇ organisms, such as bacteria or yeast cells, transformed by engineering in a manner such as to form the desired polypeptide as a fermentation product.
  • the desired polypeptide In fermentation of a substrate by means of the transformed microorganisms the desired polypeptide is in certain cases secreted to the medium.
  • the polypeptide will occur in a low concentration, e.g. 10-200 mg/1, and will be mixed with proteolytic enzymes which have simultaneous ⁇ ly been secreted from the microorganism.
  • the fermenta- tion is usually carried out at a relatively high tempera ⁇ ture, such as from 25-40°C, with a consequent risk of pro ⁇ teolytic degradation as well as de ⁇ aturation of the poly ⁇ peptide.
  • the present invention is based on the finding that the po ⁇ lypeptide can be extracted directly from the fermentation broth under specific conditions, if desired even during the very fermentation.
  • the process of the invention has been found to be useful in the recovery to proinsulin or other insulin-like material, called ILM, from a fermentation broth formed by cultivation of a correspondingly transformed yeast cell.
  • the fermentation broth was passed through an ultrafiltra ⁇ tion system 5 of the Millipore Pellican Casette Svstem type with an 0.5 ⁇ diaphragm.
  • the retentate consisting of a concentrated suspension of the yeast cells, was re ⁇ cycled through the pipe 6 to the fermentation container 1.
  • 25 1 of a human proinsulin-producing yeast culture (Sacc- haromyces cerevisiae, AB 103-1 containing the plasmid py- BCA 5) were cultivated to an optical density of 4.3 at 37°C, measured at 610 n in the fermentation container 1.
  • the fermentation broth contained human semi-synthetic insulin in a concentration of 100 mg/1.
  • the fermentation broth including yeast cells and insulin, was passed through an ultrafiltration apparatus 5 of the Pellicon Casette System type with an 0.5 ⁇ diaphragm and recycled to the fermentation container 1.
  • the yeast suspension - cultivated to an optical density of 17 - was applied from below and up through a 1.6 x 5 cm column containing suspended phenyl sepharose particles to form a fluidized bed.
  • flushing was performed with 65 ml of an equilibration buffer, and then the flow was stopped to sediment the gel.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Sustainable Development (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicinal Chemistry (AREA)
  • Endocrinology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Mycology (AREA)
  • Diabetes (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Dans un procédé de récupération d'un polypeptide, tel qu'une substance analogue à l'insuline ou hGH, contenu dans un bouillon de fermentation, on fait passer le bouillon de fermentation à travers un lit d'un matériau chromatographique contenant des groupes hydrophobes, tels que la phénylsepharose. Le matériau d'insuline adsorbé est ensuite élué avec un milieu aqueux qui peut contenir, si voulu, un solvant organique miscible avec de l'eau. La séparation chromatographique dans ces conditions permet d'obtenir un grand rendement de polypeptides isolés du bouillon de fermentation et des autres composants contenus dans le bouillon par une seule étape de traitement. Il en résulte également un facteur élevé de concentration. Un appareil pour appliquer le procédé comprend un récipient de fermentation (1) et au moins une colonne chromatographique (8, 9) connectée au récipient, de même que des dispositifs (2, 7, 10) pour faire circuler le bouillon de fermentation du récipient de fermentation (1) à travers la ou les colonnes chromatographiques (8, 9).
PCT/DK1985/000062 1984-06-25 1985-06-25 Procede et appareil de recuperation d'un polypeptide dans un bouillon de fermentation Ceased WO1986000339A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
DK84386A DK84386D0 (da) 1984-06-25 1986-02-24 Fremgangsmaade og apparat til udvinding af et polypeptid fra en fermenteringsvaeske

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DK309184A DK309184D0 (da) 1984-06-25 1984-06-25 Fremgangsmaade til isolering af insulin eller insulinlignende materiale fra en fermenteringsvaeske
DK3091/84 1984-06-25

Publications (1)

Publication Number Publication Date
WO1986000339A1 true WO1986000339A1 (fr) 1986-01-16

Family

ID=8118989

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK1985/000062 Ceased WO1986000339A1 (fr) 1984-06-25 1985-06-25 Procede et appareil de recuperation d'un polypeptide dans un bouillon de fermentation

Country Status (5)

Country Link
EP (1) EP0185078A1 (fr)
JP (1) JPS61502585A (fr)
AU (1) AU4607985A (fr)
DK (2) DK309184D0 (fr)
WO (1) WO1986000339A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0452240A1 (fr) * 1990-04-13 1991-10-16 Britech S.A. Procédé et dispositif de fermentation semi-continue, notamment pour la fabrication industrielle d'un mélange biologique à base d'acide propionique
WO1999025726A1 (fr) * 1997-11-14 1999-05-27 New Jersey Institute Of Technology Procede et appareil d'isolation et de purification de biomolecules
KR100514461B1 (ko) * 2001-09-28 2005-09-13 로레알 실란 라디칼로 치환된 파라-아미노페놀 또는파라-페닐렌디아민 화합물을 함유하는 염색 조성물
WO2006051554A1 (fr) * 2004-11-09 2006-05-18 Usv Limited Nouveau procede pour la purification d'une hormone de croissance humaine
WO2014073967A1 (fr) * 2012-11-09 2014-05-15 Biosana Pty Ltd. Traitement par lots à alimentation discontinue au moyen de bioréacteurs alternatifs
EP2698423A4 (fr) * 2011-04-14 2014-12-10 Gs Caltex Corp Appareil et procédé de séparation et de raffinage de fermentation de produit fabriqué par fermentation de micro-organismes à l'aide d'adsorbant
EP2649016A4 (fr) * 2010-12-06 2016-08-10 Tarpon Biosystems Inc Procédés de traitement continu pour produits biologiques
CN108840922A (zh) * 2018-06-04 2018-11-20 河北常山生化药业股份有限公司 分离白蛋白非结合物、白蛋白结合物和小分子化合物的方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3232531A (en) * 1963-05-13 1966-02-01 Jr Richard G Hodge Calculator

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3960720A (en) * 1973-03-23 1976-06-01 Exploaterings Aktiebolaget T.B.F. Gel product for separation purposes and method of using the product for hydrophobic salting out adsorption
EP0092280A1 (fr) * 1982-04-21 1983-10-26 Akzo N.V. Procédé semi-synthétique de préparation de l'insuline

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3960720A (en) * 1973-03-23 1976-06-01 Exploaterings Aktiebolaget T.B.F. Gel product for separation purposes and method of using the product for hydrophobic salting out adsorption
EP0092280A1 (fr) * 1982-04-21 1983-10-26 Akzo N.V. Procédé semi-synthétique de préparation de l'insuline

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Pharmacia Fine Chemicals Catalogue 1980, p 35 *
Pharmacia Fine Chemicals Catalogue 1982, p 16-17 *
Scientific American, Vol 249, 1983 Pestka, "The Purification and, Manufacture of Human Interferons". p 29-35 *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0452240A1 (fr) * 1990-04-13 1991-10-16 Britech S.A. Procédé et dispositif de fermentation semi-continue, notamment pour la fabrication industrielle d'un mélange biologique à base d'acide propionique
FR2660932A1 (fr) * 1990-04-13 1991-10-18 Britech Sa Procede et dispositif de fermentation semi-continue, notamment pour la fabrication d'un melange biologique a base d'acide propionique.
WO1991016415A1 (fr) * 1990-04-13 1991-10-31 Societe Britech S.A. Procede et dispositif de fermentation semi-continue, notamment pour la fabrication industrielle d'un melange biologique a base d'acide propionique
WO1999025726A1 (fr) * 1997-11-14 1999-05-27 New Jersey Institute Of Technology Procede et appareil d'isolation et de purification de biomolecules
KR100514461B1 (ko) * 2001-09-28 2005-09-13 로레알 실란 라디칼로 치환된 파라-아미노페놀 또는파라-페닐렌디아민 화합물을 함유하는 염색 조성물
WO2006051554A1 (fr) * 2004-11-09 2006-05-18 Usv Limited Nouveau procede pour la purification d'une hormone de croissance humaine
EP2649016A4 (fr) * 2010-12-06 2016-08-10 Tarpon Biosystems Inc Procédés de traitement continu pour produits biologiques
EP3578522A1 (fr) * 2010-12-06 2019-12-11 Pall Corporation Procédés de traitement continu pour produits biologiques
US12012629B2 (en) 2010-12-06 2024-06-18 Sartorius Stedim Chromatography Systems Ltd. Continuous processing methods for biological products
EP2698423A4 (fr) * 2011-04-14 2014-12-10 Gs Caltex Corp Appareil et procédé de séparation et de raffinage de fermentation de produit fabriqué par fermentation de micro-organismes à l'aide d'adsorbant
AU2012243504B2 (en) * 2011-04-14 2015-08-13 Gs Caltex Corporation Apparatus and method for separating and refining fermentation of product manufactured by fermenting microorganism by using adsorbent
US10150973B2 (en) 2011-04-14 2018-12-11 Gs Caltex Corporation Apparatus and method for separating and refining product manufactured by microbial fermentation by using adsorbent
WO2014073967A1 (fr) * 2012-11-09 2014-05-15 Biosana Pty Ltd. Traitement par lots à alimentation discontinue au moyen de bioréacteurs alternatifs
US9982226B2 (en) 2012-11-09 2018-05-29 Biosana Pty Ltd Discontinuous fed batch processing with the use of alternating bioreactors
CN108840922A (zh) * 2018-06-04 2018-11-20 河北常山生化药业股份有限公司 分离白蛋白非结合物、白蛋白结合物和小分子化合物的方法

Also Published As

Publication number Publication date
EP0185078A1 (fr) 1986-06-25
JPS61502585A (ja) 1986-11-13
DK84386A (da) 1986-02-24
DK309184D0 (da) 1984-06-25
DK84386D0 (da) 1986-02-24
AU4607985A (en) 1986-01-24

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