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DE4403967A1 - Process and apparatus for simultaneous multiple solid-phase synthesis - Google Patents

Process and apparatus for simultaneous multiple solid-phase synthesis

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Publication number
DE4403967A1
DE4403967A1 DE4403967A DE4403967A DE4403967A1 DE 4403967 A1 DE4403967 A1 DE 4403967A1 DE 4403967 A DE4403967 A DE 4403967A DE 4403967 A DE4403967 A DE 4403967A DE 4403967 A1 DE4403967 A1 DE 4403967A1
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Germany
Prior art keywords
reaction vessels
heating
reagents
washing
planar unit
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.)
Granted
Application number
DE4403967A
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German (de)
Other versions
DE4403967C2 (en
Inventor
Juergen Dr Haenel
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.)
Biosyntan Gesellschaft fur Bioorganische Synthese
Original Assignee
Biotez Berlin-Buch Biochemisch-Technologisches Zentrum 13125 Berlin De GmbH
Biotez Berlin Buch Bioche GmbH
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Priority to DE4403967A priority Critical patent/DE4403967C2/en
Priority to DE19944424307 priority patent/DE4424307A1/en
Publication of DE4403967A1 publication Critical patent/DE4403967A1/en
Application granted granted Critical
Publication of DE4403967C2 publication Critical patent/DE4403967C2/en
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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • 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
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00281—Individual reactor vessels
    • B01J2219/00283—Reactor vessels with top opening
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00306—Reactor vessels in a multiple arrangement
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00364—Pipettes
    • B01J2219/00367—Pipettes capillary
    • B01J2219/00369—Pipettes capillary in multiple or parallel arrangements
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00423—Means for dispensing and evacuation of reagents using filtration, e.g. through porous frits
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00479—Means for mixing reactants or products in the reaction vessels
    • B01J2219/00481—Means for mixing reactants or products in the reaction vessels by the use of moving stirrers within the reaction vessels
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00479—Means for mixing reactants or products in the reaction vessels
    • B01J2219/00484—Means for mixing reactants or products in the reaction vessels by shaking, vibrating or oscillating of the reaction vessels
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00495—Means for heating or cooling the reaction vessels
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/005—Beads
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00585—Parallel processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/0059—Sequential processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00596—Solid-phase processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00722—Nucleotides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00725—Peptides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00729—Peptide nucleic acids [PNA]
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/10—Libraries containing peptides or polypeptides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B60/00—Apparatus specially adapted for use in combinatorial chemistry or with libraries
    • C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Peptides Or Proteins (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)

Abstract

The invention relates to a process and an apparatus for fully automatic, simultaneous and multiple solid-phase synthesis of peptides, peptoids, oligonucleotides and other bioorganic polymers, and so-called libraries of these compounds. The effective support/reagent throughput achieved thereby increases the reliability of the individual syntheses, increases the purity of the crude product and permits small amounts of reagent to be used.

Description

Die vorliegende Erfindung betrifft ein Verfahren zur vollautomatischen, simultanen, multiplen Festphasensynthese und eine Vorrichtung zur Durchführung des Verfahrens. Anwendungsgebiete der Erfindung sind die chemische und die pharmazeutische Industrie und die Laborgeräteherstellung.The present invention relates to a method for fully automatic, simultaneous, multiple solid phase synthesis and a device for performing the method. Areas of application of the invention are chemical and pharmaceutical industry and laboratory equipment manufacturing.

Der Stand der Technik soll nachfolgend an Hand der simultanen, multiplen Festphasen-Peptidsynthese dargestellt werden.The state of the art is subsequently to be explained using the simultaneous multiple solid phase peptide synthesis.

Zur vollautomatischen Festphasenpeptidsynthese werden seit ca. 30 Jahren das von Merrifield entwickelte Verfahren (z. B. R.B. Merrifield, J. Am. Chem. Soc., 85 (1963) 2149; G. Barany, R.B. Merrifield in The Peptides, Analysis, Synthetics, Biology, Vol. 2, 1-284 (1980), Hrsg. Gross, Meierhofer Academy Press, New York) und weiter entwickelte Ausführungen (z. B. DE 36 31 662, DE 38 28 576, DE 40 05 518, DE 40 06 349) angewendet. Dabei wird zur Durchmischung der Reagenzien und zur Beschleunigung der Austauschreaktion zwischen fester Phase/Träger z. B. Harz und Reagenzienmischung üblicherweise das Reaktionsgefäß geschüttelt oder die Reagenzienmischung umgepumpt. Für alle Durchmi­ schungsprozesse muß hierbei in der Regel die ungenügende mechanische Stabilität der Harze berücksichtigt werden. Beim Übergang zur simultanen, multiplen Festphasen-Synthese treten in diesem Zusammenhang Probleme auf, die bisher nur ungenügend gelöst worden sind.For fully automated solid phase peptide synthesis, 30 years, the method developed by Merrifield (e.g. R.B. Merrifield, J. Am. Chem. Soc., 85 (1963) 2149; G. Barany, R.B. Merrifield in The Peptides, Analysis, Synthetics, Biology, Vol. 2, 1-284 (1980), ed. Gross, Meierhofer Academy Press, New York) and further developed versions (e.g. DE 36 31 662, DE 38 28 576, DE 40 05 518, DE 40 06 349) applied. Thereby becomes Mixing of the reagents and to accelerate the Exchange reaction between solid phase / carrier z. B. resin and Reagent mixture usually shaken the reaction vessel or pumped the reagent mixture. For all diamonds As a rule, research processes must be insufficient mechanical stability of the resins are taken into account. At the Transition to simultaneous, multiple solid phase synthesis occur this connection problems that were previously insufficient have been resolved.

Die bisher auf dem Markt erhältlichen Geräte der Firma Zinsser Analytik bzw. Abimed zur simultanen, multiplen Peptidsynthese beschreiten hierbei verschiedene Wege, um diese Probleme zu überwinden. Zum einen wird, nachdem sich das Harz abgesetzt hat, aus dem Überstand der Reagenzienmischung über dem Harz ein bestimmtes Volumen über eine Kanüle abgesaugt und gleich darauf zurückgespritzt. Dies erfordert ein zusätzliches Volumen zur Bildung des Überschusses und damit verbunden einen unnötig großen Reagenzienüberschuß. Eine ausreichende Durchmischung wird nur bei sehr geringen Harzmengen erreicht. Schon mit 20 mg Harz kann es hierbei zu Problemen bei der Durchmischung der Reagenzien kommen, und mit noch größeren Harzmengen sinkt die Effizienz dieser Durchmischungsmethode sehr schnell ab. Auf dem anderen Wege wird versucht, Lösungsmittel unterschiedlicher Dichte derart zu mischen, daß das Harz in der Mischung frei schwimmt. Nachteilig ist hier die Verringerung der aktiven Reagenzkonzentrationen durch das zweite Lösungsmittel. Das notwendige Volumen an dem zweiten Lösungsmittel, das für das freie Schwimmen des Harzes benötigt wird, ist überdies abhängig vom Harz und auch von der Peptidsequenz, ändert sich demzufolge auch während der Synthese. Folglich ist das freie Schwimmen des Harzes schwerlich in allen Fällen zu gewährleisten. Die Effizienz der Durchmischung ist auch hierbei gering und sinkt ebenfalls mit zunehmender Harzmenge.The Zinsser devices previously available on the market Analytics or Abimed for simultaneous, multiple peptide synthesis There are various ways to deal with these problems overcome. On the one hand, after the resin has settled, from the supernatant of the reagent mixture over the resin aspirated certain volume through a cannula and immediately afterwards splashed back. This requires an additional volume Formation of the surplus and associated with it unnecessarily large excess of reagents. Adequate mixing is only achieved with very small amounts of resin. Already with 20 mg  Resin can cause problems when mixing the resin Reagents are coming, and with even larger amounts of resin, it drops Efficiency of this mixing method decreases very quickly. On the other ways try different solvents Mix density so that the resin is free in the mixture swims. The disadvantage here is the reduction in active Reagent concentrations by the second solvent. The necessary volume of the second solvent for the free swimming of the resin is also dependent of the resin and also of the peptide sequence changes accordingly even during synthesis. Hence the free swimming is the Hardly to guarantee resin in all cases. The The efficiency of the mixing is also low and decreases also with increasing amount of resin.

Der Erfindung liegt die Aufgabe zugrunde, ein Verfahren und eine Vorrichtung zur simultanen, multiplen Festphasen-Synthese bereitzustellen, womit die oben genannten Nachteile vermieden werden. Diese Nachteile treffen analog auch für die Festphasensynthesen anderer bioorganischer Polymerer zu.The invention is based, a method and a task Device for simultaneous, multiple solid phase synthesis to provide, which avoids the disadvantages mentioned above become. These disadvantages also apply to the Solid phase syntheses of other bioorganic polymers.

Die Aufgabe wird durch die in den Patentansprüchen 1-9 dargelegten Maßnahmen gelöst. Die Durchmischung der Reagenzien mit dem Träger erfolgt erfindungsgemäß durch geeignete Magnetrührer in den einzelnen Reaktionsgefäßen. Eine bevorzugte Variante der Erfindung ist es, die Magnetrührer in der Harz- Reagenzien-Suspension in der Schwebe zu halte, was insbesondere bei ungenügender mechanischer Stabilität der Harze von entscheidendem Vorteil ist. Damit wird ein eventueller Abrieb der Harze, der zur Verstopfung von Fritten oder Pipetten führen kann, vermieden.The object is achieved by the in claims 1-9 outlined measures resolved. Mixing the reagents with the carrier is carried out according to the invention by suitable Magnetic stirrers in the individual reaction vessels. A preferred one Variant of the invention is the magnetic stirrer in the resin Reagent suspension in limbo, which in particular with insufficient mechanical stability of the resins from decisive advantage is. This creates a possible abrasion the resins that clog fries or pipettes can, avoided.

Die Realisierung des erfindungsgemäßen Verfahrens erfolgt durch eine Vorrichtung gemäß den Ansprüchen 10-13. Hierfür wird in einem kreisförmig angeordnetem Spulensystem unter jedem Reaktionsgefäß bzw. um jedes Reaktionsgefäß herum ein Drehfeld erzeugt. Der die Wicklungen durchfließende Strom baut ein rotierendes Magnetfeld auf, welches an einem in diesem System befindliche Magnetrührer ein Drehmoment erzeugt, wobei sich die Drehzahl des Rührers regeln läßt. Wird das in der Regel zylindrische Reaktionsgefäß in das Innere des Spulensystems gebracht, dann wird der Magnetrührer in das rotierende Magnetfeld hineingezogen, falls er sich unter dem Niveau des Magnetfelds befand, und kann sich nun freier in der Suspension bewegen. Nach dem Abschalten des Magnetfeldes sinken Rührer und Harz zu Boden und die überstehenden Lösungen können entweder von oben abpipettiert oder vorteilhafter durch eine Fritte im Boden der Reaktionsgefäße hindurch abgesaugt oder auch durch ein Schutzgas, z. B. Stickstoff, durch die Fritte hindurch abgedrückt werden.The method according to the invention is implemented by a device according to claims 10-13. For this, in a circular coil system under each Reaction vessel or a rotating field around each reaction vessel generated. The current flowing through the windings builds in rotating magnetic field, which is on one in this system  located magnetic stirrer generates a torque, the Control the speed of the stirrer. Usually it will cylindrical reaction vessel inside the coil system brought, then the magnetic stirrer is rotated into the Magnetic field if it is below the level of the Magnetic field, and can now be more freely in the suspension move. After switching off the magnetic field, stirrers and Resin to the bottom and the supernatant solutions can either be from Pipetted off at the top or more advantageously by a frit in the bottom aspirated through the reaction vessels or through a Shielding gas, e.g. B. nitrogen, pressed through the frit become.

In der einfachen Variante kann die Vorrichtung zur Durchführung des Verfahrens auch als effiziente und schonende Wasch- und Filtriereinheit, vorzugsweise für Biopolymere, trägergebundene Biopolymere, Zellsuspensionen und andere empfindliche biologische Materialien verwendet werden.In the simple variant, the device for carrying out the process also as efficient and gentle washing and Filtration unit, preferably for biopolymers, carrier-bound Biopolymers, cell suspensions and other sensitive biological materials are used.

Durch die Erfindung wird eine effektive Harz-Reagenzien- Durchmischung erreicht und dadurch die Zuverlässigkeit der individuellen Synthesen erhöht. Der geringe Anteil von Fehlsequenzen bewirkt, daß ein reineres Rohprodukt bei gleichzeitig verringertem Einsatz von Reagenzien erhalten wird. So wird beispielsweise bei der Synthese des 18-mer Peptids Monochloracetyl-GKKRPKPGGGWNTGGFRY-OH eine Ausbeutesteigerung um ca. 30% erreicht.The invention provides an effective resin reagent Mixing achieved and thereby the reliability of the individual syntheses increased. The low proportion of Incorrect sequences causes a purer raw product at the same time reduced use of reagents is obtained. For example, in the synthesis of the 18-mer peptide Monochloroacetyl-GKKRPKPGGGWNTGGFRY-OH increased the yield approx. 30% reached.

Die Erfindung soll nachfolgend durch ein Ausführungsbeispiel näher erläutert werden, durch das die Erfindung aber nicht eingeschränkt wird.The invention is intended to be described below using an exemplary embodiment are explained in more detail, but not by the invention is restricted.

AusführungsbeispielEmbodiment

Eine Stahlplatte mit den Abmessungen Breite 195 × Länge 360 × Höhe 35 mm verfügt über 5 × 10 = 50 durchgehende Bohrungen.A steel plate with the dimensions width 195 × length 360 × Height 35 mm has 5 × 10 = 50 through holes.

Die Bohrungen weisen einen Durchmesser von 18 mm zueinander auf und befinden sich im Abstand von 30 mm zueinander. Die Spulen zur Erzeugung des Drehfeldes sind kreisförmig um jedes Bohrloch unterhalb der Stahlplatte angeordnet.The holes have a diameter of 18 mm to each other and are at a distance of 30 mm from each other. The spools to generate the rotating field are circular around each borehole  arranged below the steel plate.

Die Haltevorrichtung für die Reaktionsgefäße mit einem Außendurchmesser von 16 mm befindet sich oberhalb der Stahlplatte. Die Reaktionsgefäße werden so angeordnet, daß sich ihr Boden in Höhe der Oberkante der Stahlplatte, im Innern der planaren Einheit oder ca. 5 mm unterhalb der planaren Einheit befindet.The holding device for the reaction vessels with a Outside diameter of 16 mm is above the Steel plate. The reaction vessels are arranged so that their floor at the top of the steel plate, inside the planar unit or about 5 mm below the planar unit.

Claims (14)

1. Verfahren zur Herstellung von Peptiden, Peptoiden, Oligonukleotiden und anderen bioorganischen Polymeren sowie sogenannten Bibliotheken (Librarys) dieser Verbindungen durch vollautomatische, simultane und multiple Festphasensynthese, dadurch gekennzeichnet, daß
  • - die individuellen Träger-Reagenzien-Mischungen bzw. Träger- Wasch-Mischungen in allen Reaktionsgefäßen dauernd oder zeitweilig gerührt werden,
  • - die Verteilung der Reagenzien bzw. der Waschflüssigkeiten mittels eines Pipettier-Roboters erfolgt und
  • - gegebenenfalls eine Erwärmung und/oder Vibration der Reaktionsgefäße veranlaßt wird.
1. A process for the preparation of peptides, peptoids, oligonucleotides and other bioorganic polymers and so-called libraries of these compounds by fully automatic, simultaneous and multiple solid phase synthesis, characterized in that
  • the individual carrier-reagent mixtures or carrier-washing mixtures are stirred continuously or temporarily in all reaction vessels,
  • - The reagents or washing liquids are distributed using a pipetting robot and
  • - If necessary, heating and / or vibration of the reaction vessels is caused.
2. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß der Rührvorgang durch einen Magnetrührer bewirkt wird, der in der Schwebe gehalten wird.2. The method according to claim 1, characterized in that the Stirring is effected by a magnetic stirrer in the Is suspended. 3. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß der Rührvorgang durch einen Magnetrührer bewirkt wird, der sich am Boden des Reaktionsgefäßes bewegt.3. The method according to claim 1, characterized in that the Stirring is effected by a magnetic stirrer, which is on Bottom of the reaction vessel moves. 4. Verfahren nach Anspruch 1-3, dadurch gekennzeichnet, daß die Bewegungen der Magnetrührer durch rotierende Magnetfelder in kreisförmig angeordneten Spulensystemen hervorgerufen werden.4. The method according to claim 1-3, characterized in that the Movements of the magnetic stirrer through rotating magnetic fields circular coil systems are caused. 5. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß die Reagenzien bzw. Waschflüssigkeiten durch eine oder mehrere Nadeln eines Pipettier-Roboters abgesaugt werden.5. The method according to claim 1, characterized in that the Reagents or washing liquids by one or more Needles of a pipetting robot are sucked off. 6. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß die Reagenzien bzw. Waschflüssigkeiten gleichzeitig aus allen Reaktionsgefäßen durch eine im Boden befindliche Filterfritte entfernt werden, vorzugsweise durch Anlegen eines Vakuums oder der Anwendung eines mit einem Schutzgas erzeugten Überdrucks. 6. The method according to claim 1, characterized in that the Reagents or washing liquids from all at the same time Reaction vessels through a filter frit located in the bottom are removed, preferably by applying a vacuum or the application of an excess pressure generated with a protective gas.   7. Verfahren nach Anspruch 1-4, dadurch gekennzeichnet, daß eine Temperierung der Reaktionsgefäße auf 0-120°C, vorzugsweise auf 30-50°C, erfolgt.7. The method according to claim 1-4, characterized in that a Temperature control of the reaction vessels to 0-120 ° C, preferably to 30-50 ° C. 8. Verfahren nach Anspruch 1-4, dadurch gekennzeichnet, daß die Erwärmung der Reaktionsgefäße durch Heizspulen, geeignete Flächenheizungen (Heizfolien) oder Thermostaten, erfolgt.8. The method according to claim 1-4, characterized in that the Heating the reaction vessels using heating coils, suitable Surface heating (heating foils) or thermostats. 9. Verfahren nach Anspruch 1-4, dadurch gekennzeichnet, daß die Vibration der Reaktionsgefäße durch Ultraschall veranlaßt wird.9. The method according to claim 1-4, characterized in that the Vibration of the reaction vessels is caused by ultrasound. 10. Vorrichtung zur Durchführung des Verfahrens nach Anspruch 1-9, gekennzeichnet durch in einer planaren Einheit zusammengefaßte, kreisförmig angeordnete Spulensysteme, die sich oberhalb oder unterhalb der in einer Haltevorrichtung fixierten Reaktionsgefäße befindet.10. Apparatus for performing the method according to claims 1-9, characterized by in a planar unit summarized, circularly arranged coil systems, which above or below that fixed in a holding device Reaction vessels. 11. Vorrichtung nach Anspruch 10, gekennzeichnet durch in der planaren Einheit angeordnete Heizvorrichtungen gemäß Anspruch 8.11. The device according to claim 10, characterized by in the A planar unit heater according to claim 8. 12. Vorrichtung nach Anspruch 10-11, gekennzeichnet durch senkrechte Bohrungen in der planaren Einheit im jeweiligen Mittelpunkt der kreisförmig angeordneten Spulensysteme.12. The apparatus according to claim 10-11, characterized by vertical holes in the planar unit in each Center of the circularly arranged coil systems. 13. Vorrichtung nach Anspruch 10-12, gekennzeichnet durch eine Positionierung des Bodens der Reaktionsgefäße oberhalb, innerhalb oder unterhalb der planaren Einheit.13. The apparatus according to claim 10-12, characterized by a Positioning the bottom of the reaction vessels above, inside or below the planar unit. 14. Verwendung der Vorrichtung nach Anspruch 10 bis 13 als Wasch- und Filtriereinheit, vorzugsweise für Biopolymere, trägergebundene Biopolymere, Zellsuspensionen und andere empfindliche biologische Materialien.14. Use of the device according to claim 10 to 13 as Washing and filtering unit, preferably for biopolymers, carrier-bound biopolymers, cell suspensions and others sensitive biological materials.
DE4403967A 1993-02-10 1994-02-04 Method and device for producing peptides, peptoids and oligonucleotides and their use Expired - Fee Related DE4403967C2 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5792430A (en) * 1996-08-12 1998-08-11 Monsanto Company Solid phase organic synthesis device with pressure-regulated manifold

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US4465377A (en) * 1983-06-07 1984-08-14 Techne Corporation Magnetic stirrer apparatus with guided, floating stirrer
US4498785A (en) * 1982-06-09 1985-02-12 Techne Corporation Floating magnetic stirrer for culture medium
US4759635A (en) * 1985-08-08 1988-07-26 Techne Corporation Magnetic stirrer apparatus with improved stirring action

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US4498785A (en) * 1982-06-09 1985-02-12 Techne Corporation Floating magnetic stirrer for culture medium
US4465377A (en) * 1983-06-07 1984-08-14 Techne Corporation Magnetic stirrer apparatus with guided, floating stirrer
US4759635A (en) * 1985-08-08 1988-07-26 Techne Corporation Magnetic stirrer apparatus with improved stirring action

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5792430A (en) * 1996-08-12 1998-08-11 Monsanto Company Solid phase organic synthesis device with pressure-regulated manifold

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