EP0417227A1 - Procede biocatalytique avec des particules porteuses sous forme de particules magnetiques de verre ou de ceramique, et dispositif pour son execution - Google Patents

Procede biocatalytique avec des particules porteuses sous forme de particules magnetiques de verre ou de ceramique, et dispositif pour son execution

Info

Publication number
EP0417227A1
EP0417227A1 EP19900904292 EP90904292A EP0417227A1 EP 0417227 A1 EP0417227 A1 EP 0417227A1 EP 19900904292 EP19900904292 EP 19900904292 EP 90904292 A EP90904292 A EP 90904292A EP 0417227 A1 EP0417227 A1 EP 0417227A1
Authority
EP
European Patent Office
Prior art keywords
permanent magnet
particles
carrier particles
reactor
glass
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.)
Withdrawn
Application number
EP19900904292
Other languages
German (de)
English (en)
Inventor
Ralf Kindervater
Rolf Dieter Schmid
Wolfgang Dr. Künneker
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.)
KINDERVATER Ralf
Original Assignee
Helmholtz Zentrum fuer Infektionsforschung HZI GmbH
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 Helmholtz Zentrum fuer Infektionsforschung HZI GmbH filed Critical Helmholtz Zentrum fuer Infektionsforschung HZI GmbH
Publication of EP0417227A1 publication Critical patent/EP0417227A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G01N33/5434—Magnetic particles using magnetic particle immunoreagent carriers which constitute new materials per se
    • 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
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2446/00—Magnetic particle immunoreagent carriers
    • G01N2446/10—Magnetic particle immunoreagent carriers the magnetic material being used to coat a pre-existing polymer particle but not being present in the particle core
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2446/00—Magnetic particle immunoreagent carriers
    • G01N2446/80—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids
    • G01N2446/90—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids characterised by small molecule linker used to couple immunoreagents to magnetic particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation

Definitions

  • Biocatalytic process and carrier particles consisting of magnetic glass or ceramic particles and device for carrying them out.
  • biocatalysts such as proteins, DNA structures or microorganisms
  • the chemical or biological substance can also be measured indirectly by removing the substance or the reaction products from the reaction site and then allowing a further reaction to take place at the reaction site with which the course of the first reaction can be measured. This second reaction can thus be carried out without any interference from starting materials or products of the first reaction.
  • CPG porous glass
  • Chen modification have been subjected, for example with the help of aminopropylsilane.
  • Enzymes can be bound, for example, to such surface-modified CPG using the known Carbodii id, thiourea or azo method. Those skilled in the art are familiar with these techniques; see. for example Filbert, Controlled-Pore Glasses for Enzyme Immobilization. In: Messing, I mobilized Enzymes for Industrial Reactors, 39-62, NY, 1975.
  • a catalytic process which runs in liquid, in particular aqueous media, is now proposed, in which a biocatalyst is used which is fixed on glass or ceramic particles, in particular on porous glass (CPG) as a support, the process thereby resulting in is characterized in that the carrier particles are magnetic or magnetizable.
  • CPG porous glass
  • Such particles can be held in the liquid medium with the help of a magnetic field. If the process is carried out in a reactor through which the liquid medium flows, the carrier particles can easily be removed by switching off the magnetic field.
  • biocatalysts As in the prior art, biological macromolecules, such as antibodies, lectins, enzymes or DNA structures, organisms, such as microorganisms, or parts thereof, such as animal or plant tissue, are suitable as biocatalysts.
  • carrier particles which may be surface-modified ceramic material or glass, in particular porous glass (CPG), on which a fur / Felll mixed oxide / mixed hydroxide has been deposited.
  • a device which is characterized by a reactor for carrying out the catalytic process, a permanent magnet and an electromagnet, the reactor being arranged in the field of the permanent magnet and the permanent magnet using the Electromagnets can be switched off.
  • the device according to the invention can be used
  • the cover being a magnetizable element for the force connection between the poles of the permanent magnet and
  • FIG. 1 shows a schematic of a flow injection analysis (FIA);
  • FIG. 2 shows a device according to the invention for carrying out an FIA;
  • FIGS. 3 and 4 show further embodiments of the device according to the invention.
  • a water sample is to be examined for impurities, for example inhibitors of cholinesterase.
  • the water sample is included With the help of one of the pumps of the pump block 4 to 6 via the line 3 and the valve 12, it is fed to a reactor 17 in which the impurity sought is subjected to a reaction with the aid of a biocatalyst.
  • the reactor can be a reaction or line loop in a permanent magnet system which can be switched off and in which magnetizable CPGs are held by means of the magnetic field, on which, for example, cholinesterase has been immobilized.
  • the immobilized choline esterase is gradually poisoned by the impurity sought.
  • the outflow takes place via the valve 13 and the line 20.
  • the valve 12 is actuated so that the inflow of the water sample via line 3 is interrupted and the reactor via line 1 with the help of one of the pumps of pump block 4 to 6 a substrate is fed which is subjected to a reaction by the not yet inactivated biocatalyst.
  • the substrate is choline ester.
  • the substrate supplied to the reactor 17 via the line 1 and the valves 11 and 12 can be diluted with a liquid carrier which meets the substrate in the valve 11.
  • the liquid carrier can be a buffer solution.
  • the stream leaving the reactor 17 is fed via the valve 13 to a secondary reactor 21, in which the reaction product of the biocatalytic reaction is subjected to a further reaction, the result of which can be determined in the downstream detector 22.
  • the subsequent reaction is the oxidation of the choline released in the reactor 17 with the aid of choline oxidase, which produces hydrogen peroxide, which can be measured in the detector 22.
  • Carrier with unused biocatalyst is kept ready in the storage vessel 14 and can be stirred there with the aid of a stirrer 15 slurried. If the biocatalyst in the reactor 17 is used up, the permanent magnet (not shown) can be switched off, so that the used biocatalyst can be discharged via the valve 13 and the line 20 with the aid of the carrier stream 2. Unused biocatalyst can be fed to the reactor 17 from the storage vessel 14 via the line 16 and the valve 12.
  • the reactor 17 (FIG. 1) is shown in more detail in FIG. 2.
  • the permanent magnet 100 is formed like a gugelhupf, so that its two poles 101, 102 are formed by the edge of the ring wall 101 and the central pin 102.
  • This permanent magnet 100 is additionally provided with an electromagnet, of which only the connections 103, 104 can be seen.
  • the permanent magnet 100 can be switched off by the electromagnet, not shown.
  • the permanent magnet 100 carries a cover 105 made of a magnetically neutral material, for example brass. This cover 105 is provided on its side facing the permanent magnet 100 with a steel disk 106 which has a smaller diameter than the cover 105 and which produces the force fit between the poles 101 and 102.
  • a feed line 118 is brought to the periphery of the steel disk 106 and, as the actual reactor 117, runs around the steel disk 106 and then leads away as a line 119.
  • the actual reactor is thus placed as an approximately omega-shaped conductor loop 117 around the steel disk 106 and can lie on the periphery of the steel disk 106 in a groove in the cover 105.
  • 3 shows a further embodiment of a device according to the invention using a module.
  • the module consists of a base plate on which the coil of an electromagnet with a circular core is mounted so that a hose is guided between a gap in the core with tapered ends, which transports the magnetic particles.
  • a block made of non-magnetic material and fitted with a device for receiving a hose serves as the hose holder.
  • FIG. 4 shows a further embodiment in the form of a device according to the invention on the basis of a further module.
  • the module consists of a base plate on which a permanent magnet can be moved back and forth in the longitudinal direction by an actuator.
  • a block made of non-magnetic material which is firmly attached to the base plate and is equipped with a device for receiving a hose serves as the rear stop. At both ends of the block there are devices for holding the hose tight in the guide.
  • the servomotor is controlled so that the permanent magnet is moved against the stop block. So the tube through which the magnetic particles can flow comes under the direct influence of the magnetic field forces, which are concentrated in the slot of the magnet, between the two poles.
  • the magnet can be moved backwards by actuating the servomotor, so that the magnetic field acting on the hose is eliminated. She likes. Particles in the hose are no longer held and transported away in the direction of flow.
  • Cholinesterase was immobilized on the CPG particles obtained in Production Example 1 as follows. First with 5 percent. Glutaraldehyde activated in 0.1 M potassium phosphate buffer of pH 7.5. 50 mg glass and 2 ml aldehyde solution were slowly agitated in a sealed vessel for 30 minutes. The glass took on a strong pink color over the course of this time. The glass thus activated was washed six times with buffer, after which it was decanted and the supernatant was discarded. After the last washing, a solution of 10 mg / ml enzyme preparation in buffer could be added, after which the mixture was slowly rotated for 1 hour. The supernatant of the mixture was used for the Bradford protein determination. The slides were washed three times with buffer and then stored in buffer at 4C.
  • Glucose oxidase (GOD) was immobilized on CPG analogously to Example 1 with an enzyme concentration of 10 mg / ml buffer.
  • the composition of the substrate was as follows: 0.1 M potassium phosphate buffer of pH 7.5; dissolved therein per ml 21, uM glucose, 2 ⁇ uM ABTS and 10 units POD. 0.1 M potassium phosphate buffer of pH 7.5 was used as the carrier stream. The hydrogen peroxide formed by oxidation of the dye ABTS was detected photometrically at a wavelength of 420 nm.
  • Alcohol oxidase (AOD) was immobilized on CPG at an enzyme concentration of 10 mg / ml buffer according to Example 1.
  • a flow injection analysis is carried out, pulsing every 40 seconds with a substrate solution of the following composition: 0.1 M potassium phosphate buffer of pH 7.5; dissolved per ml: 300.UM ethanol, 2.uM ABTS and 10 units POD.
  • the detection in the photometer was carried out at 420 ⁇ m.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Hematology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Inorganic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé catalytique se déroulant dans un milieu fluide, et en particulier aqueux, ainsi que des particules porteuses et un dispositif pour la mise en oeuvre du procédé. Le biocatalyseur est fixé sur des particules de verre ou de céramique, en particulier sur du verre poreux, et les particules porteuses sont magnétiques. La surface des particules porteuses est recouverte d'une couche de séparation constituée d'un polymère organique, par exemple le glutaraldéhyde ou le polyuréthane.
EP19900904292 1989-03-15 1990-03-15 Procede biocatalytique avec des particules porteuses sous forme de particules magnetiques de verre ou de ceramique, et dispositif pour son execution Withdrawn EP0417227A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE3908494 1989-03-15
DE3908494 1989-03-15

Publications (1)

Publication Number Publication Date
EP0417227A1 true EP0417227A1 (fr) 1991-03-20

Family

ID=6376423

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19900904292 Withdrawn EP0417227A1 (fr) 1989-03-15 1990-03-15 Procede biocatalytique avec des particules porteuses sous forme de particules magnetiques de verre ou de ceramique, et dispositif pour son execution

Country Status (3)

Country Link
EP (1) EP0417227A1 (fr)
JP (1) JPH03505163A (fr)
WO (1) WO1990010696A1 (fr)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5397755A (en) * 1993-06-29 1995-03-14 W. R. Grace & Co.-Conn. Low density glassy materials for bioremediation supports
FR2766387B1 (fr) * 1997-07-22 1999-10-08 Univ La Rochelle Procede reactionnel en continu par catalyse solide/gaz en milieu non conventionnel, reacteur correspondant et utilisation de ce reacteur
DE19822050A1 (de) * 1998-05-16 1999-11-18 Forschungszentrum Juelich Gmbh Verfahren zur Durchführung von chemischen oder biologischen Reaktionen
EP1693387B1 (fr) * 2003-07-17 2012-06-20 Invitrogen Dynal AS Procédé de préparation de particules magnétiques revêtues
CA2532711C (fr) * 2003-07-17 2013-07-23 Dynal Biotech Asa Revetement de particules polymeres poreuses a surface fonctionnalisee qui contiennent des cristaux superparamagnetiques avec monomeres polyisocyanates/diols ou epoxides
US20060188905A1 (en) 2005-01-17 2006-08-24 Dynal Biotech Asa Process
GB0500888D0 (en) * 2005-01-17 2005-02-23 Dynal Biotech Asa Process
JP6880571B2 (ja) * 2016-05-20 2021-06-02 Jnc株式会社 磁性粒子を用いた水溶液中の微生物の回収方法および回収装置

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4233169A (en) * 1979-04-13 1980-11-11 Corning Glass Works Porous magnetic glass structure
SE456164B (sv) * 1980-08-20 1988-09-12 Kjell Nilsson Forfarande for immobilisering, odling och efterfoljande frigoring av animala celler samt mikroberare av gelatin med absorberade animala celler
US4360441A (en) * 1981-06-25 1982-11-23 Corning Glass Works Glass-encapsulated magnetic materials and methods for making them
US4448884A (en) * 1982-03-03 1984-05-15 Kms Fusion, Inc. Glass-surface microcarrier for growth of cell cultures
JPS6033476B2 (ja) * 1982-07-29 1985-08-02 ユ−オ−ピ−・インコ−ポレイテツド 磁性支持マトリツクス及び固定化酵素系
DE3228477C2 (de) * 1982-07-30 1985-01-31 Uop Inc., Des Plaines, Ill. Magnetisches immobilisiertes Enzymsystem

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9010696A1 *

Also Published As

Publication number Publication date
WO1990010696A1 (fr) 1990-09-20
JPH03505163A (ja) 1991-11-14

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