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WO2002005945A1 - Method for producing microarray chips with nucleic acids, proteins or other test substrates - Google Patents

Method for producing microarray chips with nucleic acids, proteins or other test substrates Download PDF

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Publication number
WO2002005945A1
WO2002005945A1 PCT/DE2001/002559 DE0102559W WO0205945A1 WO 2002005945 A1 WO2002005945 A1 WO 2002005945A1 DE 0102559 W DE0102559 W DE 0102559W WO 0205945 A1 WO0205945 A1 WO 0205945A1
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WO
WIPO (PCT)
Prior art keywords
substrate
carrier substance
fibers
strips
chips
Prior art date
Application number
PCT/DE2001/002559
Other languages
German (de)
French (fr)
Inventor
Kang-Hun Lee
Original Assignee
Max-Delbrück-Centrum für Molekulare Medizin
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Publication date
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Publication of WO2002005945A1 publication Critical patent/WO2002005945A1/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/36—Embedding or analogous mounting of samples
    • 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
    • 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
    • B01J2219/00457—Dispensing or evacuation of the solid phase support
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • B01J2219/00608—DNA chips
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01J2219/00612—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports the surface being inorganic
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    • B01J2219/00614—Delimitation of the attachment areas
    • B01J2219/00621—Delimitation of the attachment areas by physical means, e.g. trenches, raised areas
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    • B01J2219/00623—Immobilisation or binding
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Definitions

  • the human genome project which is about to be fully decoded, is currently undergoing a drastic change in the biomedical field.
  • the so-called microarray technology appears to be the first technical novelty that has the potential to be comparable to PCR technology in the 1980s (see “insighf” article in the Nature edition of June 15, 2000, vol. 405, from page 819: Functional genomics).
  • the production of so-called microarray chips for biomedical research is mainly implemented by two methods: 1.
  • the substrates DNA or protein
  • the dots are applied through nozzles which are adapted from the nozzles of inkjet printers.
  • the object of the present invention is a new method for the production of microarray chips.
  • the main objectives of the invention are a low variability of the chips (quality improvement) and a higher economy in the production, especially of large quantities of the same chips.
  • the object of the invention is achieved according to the claims.
  • the inventive method is characterized in that
  • a substrate e.g. DNA, protein or chemical substance
  • a carrier substance e.g.
  • Paraffin, polyacrylamide Paraffin, polyacrylamide
  • Micrometer range is cast, shaped or cut, various fibrous carrier substances with the respective substrate in any
  • Composition are combined into a bundle, the individual fibers are isolated from each other in the bundle by separating material, the combination of different fibers is brought in a defined arrangement, alternatively the carrier substance with the substrate is poured, shaped or cut in layers, different layers are stacked in layers and separating layers separate layers with different substrates, the entire layer is cut into strips, different strips are put together to form a block, again
  • Separating layers are integrated between strips, the different layers and strips are produced in a defined arrangement, the separating material or the separating layer consists of the same carrier material without a substrate, the separating material or the separating layer consists of a different material, which
  • Carrier material adheres, the bundles or blocks are cut into thin slices of a few micrometers, the slices are applied to a suitable solid surface, the carrier substance is removed from the surface and the substrate is immobilized at the respective location on the surface, alternatively: the carrier substance is not removed, but the substrate on the surface of the
  • the substrate is homogeneously distributed in a carrier substance as the first step.
  • a carrier substance A wide variety of test substances can be used as a substrate, which are available in large numbers, in particular DNA for gene expression analyzes and proteins for proteome analyzes, but also chemical substances, such as those being investigated in the search for new drugs.
  • a wide variety of substances can be used as the carrier substance, which enable a homogeneous mixture of the substrate and do not damage the substrate.
  • the carrier substance can first be shaped, is then converted into a solid state either chemically (by a chemical reaction, for example polyacrylamide gel) or physically (by cooling, for example paraffin).
  • the carrier substance with the substrate contained therein is cast, drawn or cut in the form of microscopic fibers (up to diameters of less than 100 micrometers). Fibers with different substrates are bundled in a defined arrangement. Separating material separates different fibers so that the substrates are isolated from each other. Alternatively, the carrier substance with the substrate is applied to one another in layers (up to thicknesses of less than 100 micrometers) in layers. Here, too, separating layers isolate layers with different substrates from one another. The entire layer is cut into fine strips (up to widths of less than 100 micrometers). Different strips are combined into a block. The separating material or the separating layer either consists of the same carrier material without a substrate or of a different material that can be adhered to the carrier material.
  • the fiber bundle or strip block is cut across the longitudinal axis into thin slices (up to thicknesses of less than 100 microns), placed on a firm surface and fixed.
  • Glass plates, metal plates or other plates which are suitable for immobilizing the substrate covalently or non-covalently are used as supports.
  • the discs are fixed chemically (e.g. by covalently binding the separating material to the chemically pretreated surface) or physically (e.g. by gluing, pressing or burning).
  • the carrier substance is removed and the substrate is immobilized on the support.
  • the carrier substance is left, for this purpose the substrate is brought from the carrier substance to the surface (e.g. by an electric field with a polyacrylamide gel as carrier substance) and is thus fixed.
  • microarray chips with substrates in the order of 10 6 and more can be produced in a series of 1000 to 10 6 pieces.
  • the number and dimensions of the substrate points correspond to the dimension in the case of chips currently being produced by others
  • DNA microchips for example, have already found their way into research institutes and the pharmaceutical industry, and further areas of application are opening up in rapid succession. A potentially even larger market is expected to be in clinical use for screening tests.
  • the previous manufacturing techniques are relatively inefficient, i.e. measured by the financial and time expenditure, the production is low. As a result, demand is already greater than supply, and this will intensify in the coming years, especially after the completion of the human genome project. It is even more important that the conventional chips are very expensive due to the high technical complexity, making the chips financially unaffordable for many applications that would make scientific and medical sense. The process presented here will be able to drastically reduce production costs and thus prices, while at the same time achieving a qualitative improvement in the chips.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention relates to the production of microarray chips for analyzing a large number of individual values, especially DNA or protein microarray chips. According to said method, the substrate (e.g. DNA, protein or other test substances) is homogeneously distributed in a support substance. The support substance, together with the substrate content therein, is transformed into microscopically thin fibers and different fibers are arranged in a bundle. Alternatively, the support substance and the substrate are applied to each other in layers, the combination layer is cut into strips and different strips are combined to form a block. The bundle or block is cut into thin slices crosswise to the longitudinal axis, applied to a solid support and fixed. The support substance is removed and the substrate is immobilized on said support. This method enables microarray chips to be produced with as many substrates as required. The outstanding advantages of the method are as follows: 1. simple production of an almost unlimited number of chips (one bundle/block 10 cm in length produces 1000 slices = chips 100 micrometers thick); 2. very little variation among the chips in a series (the 1000 slices are almost identical in terms of substrate content). The invention hereby provides an efficient and economical means of producing microarray chips for the increasing demand in biomedical research and industry.

Description

Verfahren zur Herstellung von Mikroarray-Chips mit Nukleinsäuren, Proteinen oder anderen TestsubstratenProcess for the production of microarray chips with nucleic acids, proteins or other test substrates
Beschreibungdescription
Nicht zuletzt mit dem Humanen Genomprojekt, das kurz vor der vollständigen Entschlüsselung der menschlichen Erbinformation steht, findet zur Zeit ein drastischer Umbruch im biomedizinischen Feld statt. In diesem Zusammenhang scheint die sogenannte Mikroarray-Technik als erste technische Neuheit hervorzugehen, die ein Potenzial hat vergleichbar der PCR-Technik in den 80er Jahren (siehe „insighf'-Artikel der Nature- Ausgabe vom 15 Juni 2000, Vol.405, ab Seite 819: Functional genomics). Gegenwärtig wird die Herstellung von sogenannten Mikroarray-Chips für die biomedizinische Forschung vor allem durch zwei Verfahren realisiert: 1. Auf einer Unterlage (Glas oder Membran) werden die Substrate (DNA oder Protein) als Punkte durch mechanisch betriebene feine Metallspitzen aufgetragen; 2. Das Auftragen der Punkte geschieht durch Düsen, die aus den Düsen von Tintenstrahldruckern adaptiert sind.Last but not least, the human genome project, which is about to be fully decoded, is currently undergoing a drastic change in the biomedical field. In this context, the so-called microarray technology appears to be the first technical novelty that has the potential to be comparable to PCR technology in the 1980s (see "insighf" article in the Nature edition of June 15, 2000, vol. 405, from page 819: Functional genomics). At present, the production of so-called microarray chips for biomedical research is mainly implemented by two methods: 1. The substrates (DNA or protein) are applied as dots on a base (glass or membrane) using mechanically operated fine metal tips; 2. The dots are applied through nozzles which are adapted from the nozzles of inkjet printers.
Beide Verfaliren sind zwar in der Lage, Chips mit bis zu Millionen von individuellen Punkten von ca. 10-100 Mikrometer Durchmesser herzustellen, jedoch die Herstellung ist aufwendig aufgrund des Herstellungsprinzips, dass jeder Punkt auf jedem Chip einzeln aufgebracht werden muss. Dieser Nachteil nimmt proportional mit der Anzahl der Substrate (Punkte pro Chip) und der Anzahl der Chips zu. Ein weiterer, qualitativer Nachteil ist die Variabilität der Punkte von Chip zu Chip sogar in derselben Herstellungsserie. Dies ist einerseits durch die mechanische Limitation der Pinspitzen der Auftragungsroboter, andererseits wieder im Herstellungsverfahren (das Auftragen von individuellen Punkten) begründet und beeinträchtigt die Vergleichbarkeit der Resultate.Both methods are able to produce chips with up to millions of individual points of approximately 10-100 micrometers in diameter, but the production is complex due to the manufacturing principle that each point must be applied to each chip individually. This disadvantage increases proportionally with the number of substrates (points per chip) and the number of chips. Another qualitative disadvantage is the variability of the points from chip to chip even in the same production series. This is due on the one hand to the mechanical limitation of the pin tips of the application robots, and on the other hand again in the manufacturing process (the application of individual points) and impairs the comparability of the results.
Vor diesem Hintergrund ist die Aufgabe der vorliegenden Erfindung ein neues Verfahren für die Herstellung von Mikroarray-Chips. Die Hauptziele der Erfindung sind eine geringe Variabilität der Chips (Qualitätsverbesserung) und eine höhere Ökonomie der Herstellung vor allem von großen Mengen gleicher Chips. Die Aufgabe der Erfindung wird gemäß den Ansprüchen realisiert. Das erfindungsgemäße Verfahren ist dadurch gekennzeichnet, daßAgainst this background, the object of the present invention is a new method for the production of microarray chips. The main objectives of the invention are a low variability of the chips (quality improvement) and a higher economy in the production, especially of large quantities of the same chips. The object of the invention is achieved according to the claims. The inventive method is characterized in that
ein Substrat (z.B. DNA, Protein oder chemische Substanz) in einer Trägersubstanz (z.B.a substrate (e.g. DNA, protein or chemical substance) in a carrier substance (e.g.
Paraffin, Polyacrylamid) homogen aufgelöst wird, die Trägersubstanz unter Bedingungen, die das enthaltene Substrat nicht zerstören, in festen Aggregatzustand überführt wird, die Trägersubstanz mit dem Substrat in eine faserförmige Form mit Durchmesser imParaffin, polyacrylamide) is homogeneously dissolved, the carrier substance is converted into a solid state under conditions which do not destroy the substrate contained, the carrier substance with the substrate in a fibrous form with a diameter in
Mikrometerbereich gegossen, geformt oder geschnitten wird, verschiedene faserförmige Trägersubstanzen mit dem jeweiligen Substrat in beliebigerMicrometer range is cast, shaped or cut, various fibrous carrier substances with the respective substrate in any
Zusammensetzung zu einem Bündel kombiniert werden, die einzelnen Fasern durch Trennmaterial voneinander isoliert im Bündel vorliegen, die Kombination von verschiedenen Fasern in definierter Anordnung gebracht werden, alternativ die Trägersubstanz mit dem Substrat in Schichten gegossen, geformt oder geschnitten wird, verschiedene Schichten in Lagen übereinander geschichtet werden und Trennschichten jeweils Schichten mit unterschiedlichen Substraten separieren, die Gesamtschicht in Streifen geschnitten wird, verschiedene Streifen zu einem Block zusammengesetzt werden, wobei wiederumComposition are combined into a bundle, the individual fibers are isolated from each other in the bundle by separating material, the combination of different fibers is brought in a defined arrangement, alternatively the carrier substance with the substrate is poured, shaped or cut in layers, different layers are stacked in layers and separating layers separate layers with different substrates, the entire layer is cut into strips, different strips are put together to form a block, again
Trennschichten zwischen Streifen integriert werden, die verschiedenen Schichten und Streifen in definierter Anordnung hergestellt werden, das Trennmaterial bzw. die Trennschicht aus demselben Trägermaterial ohne Substrat besteht, das Trennmaterial bzw. die Trennschicht aus einem anderen Material besteht, das amSeparating layers are integrated between strips, the different layers and strips are produced in a defined arrangement, the separating material or the separating layer consists of the same carrier material without a substrate, the separating material or the separating layer consists of a different material, which
Trägermaterial haftet, die Bündel oder die Blöcke in dünnen Scheiben von wenigen Mikrometern geschnitten werden, die Scheiben auf einer geeigneten festen Unterlage aufgetragen werden, die Trägersubstanz von der Unterlage entfernt wird und dabei das Substrat an der jeweiligen Stelle auf der Unterlage immobilisiert wird, alternativ: die Trägersubstranz nicht entfernt, jedoch das Substrat auf die Oberfläche derCarrier material adheres, the bundles or blocks are cut into thin slices of a few micrometers, the slices are applied to a suitable solid surface, the carrier substance is removed from the surface and the substrate is immobilized at the respective location on the surface, alternatively: the carrier substance is not removed, but the substrate on the surface of the
Scheibe gebracht und mitsamt der Trägersubstanz auf der Unterlage immobilisiert wird. Bei dem erfindungsgemäßen Verfahren wird als erster Schritt das Substrat in einer Trägersubstanz homogen verteilt. Als Substrat können verschiedenste Testsubstanzen verwendet werden, die in großer Anzahl vorliegen, insbesondere DNS für Genexpressionsanalysen und Proteine für Proteomanalysen, aber auch chemische Substanzen, wie sie bei der Suche nach neuen Medikamenten untersucht werden. Als Trägersubstanz können verschiedenste Substanzen verwendet werden, die eine homogene Mischung des Substrats ermöglichen und nicht das Substrat beschädigen. Die Trägersubstanz ist zunächst formbar, wird dann entweder chemisch (durch eine chemische Reaktion, z.B. Polyacrylamidgel) oder physikalisch (durch Abkühlung, z.B. Paraffin) in einen festen Zustand überfuhrt.Brought disk and immobilized on the base together with the carrier. In the method according to the invention, the substrate is homogeneously distributed in a carrier substance as the first step. A wide variety of test substances can be used as a substrate, which are available in large numbers, in particular DNA for gene expression analyzes and proteins for proteome analyzes, but also chemical substances, such as those being investigated in the search for new drugs. A wide variety of substances can be used as the carrier substance, which enable a homogeneous mixture of the substrate and do not damage the substrate. The carrier substance can first be shaped, is then converted into a solid state either chemically (by a chemical reaction, for example polyacrylamide gel) or physically (by cooling, for example paraffin).
Die Trägersubstanz mit dem darin enthaltenen Substrat wird in Form von mikroskopisch dünnen Fasern (bis zu Durchmessern von unter 100 Mikrometer) gegossen, gezogen oder geschnitten. Fasern mit unterschiedlichen Substaten werden in definierter Anordung gebündelt. Trennmaterial separiert verschiedene Fasern, so dass die Substrate voneinander isoliert sind. Alternativ wird die Trägersubstanz mit dem Substrat in dünnen Schichten (bis zu Dicken von unter 100 Mikrometer) in Lagen aufeinander aufgetragen. Auch hier isolieren Trennschichten Lagen mit verschiedenen Substraten voneinander. Die Gesamtschicht wird in feinen Streifen (bis zu Breiten von unter 100 Mikrometer) geschnitten. Verschiedene Streifen werden zu einem Block vereint. Das Trennmaterial bzw. die Trennschicht besteht entweder aus demselben Trägermaterial ohne Substrat oder aus einem anderen Material, das sich an das Trägermaterial haften lässt.The carrier substance with the substrate contained therein is cast, drawn or cut in the form of microscopic fibers (up to diameters of less than 100 micrometers). Fibers with different substrates are bundled in a defined arrangement. Separating material separates different fibers so that the substrates are isolated from each other. Alternatively, the carrier substance with the substrate is applied to one another in layers (up to thicknesses of less than 100 micrometers) in layers. Here, too, separating layers isolate layers with different substrates from one another. The entire layer is cut into fine strips (up to widths of less than 100 micrometers). Different strips are combined into a block. The separating material or the separating layer either consists of the same carrier material without a substrate or of a different material that can be adhered to the carrier material.
Das Faserbündel bzw. der Streifenblock wird quer zur Längsachse in dünne Scheiben (bis zu Dicken von unter 100 Mikrometer) geschnitten, auf eine feste Unterlage gebracht und fixiert. Als Unterlagen werden Glasplatten, Metallplatten oder sonstige Platten verwendet, die geeignet sind, das Substrat kovalent oder nicht-kovalent zu immobilisieren. Die Fixation der Scheiben erfolgt chemisch (z.B. durch kovalente Bindung des Trennmaterial an die chemisch vorbehandelte Unterlage) oder physikalisch (z.B. durch Kleben, Pressen oder Aufbrennen). Während oder nach der Fixierung wird die Trägersubstanz entfernt und das Substrat auf der Unterlage immobilisiert. Alternativ wird die Trägersubstanz belassen, dafür das Substrat aus der Trägersubstanz an die Oberfläche gebracht (z.B. durch ein elektrisches Feld bei einem Polyacrylamidgel als Trägersubstanz) und so fixiert.The fiber bundle or strip block is cut across the longitudinal axis into thin slices (up to thicknesses of less than 100 microns), placed on a firm surface and fixed. Glass plates, metal plates or other plates which are suitable for immobilizing the substrate covalently or non-covalently are used as supports. The discs are fixed chemically (e.g. by covalently binding the separating material to the chemically pretreated surface) or physically (e.g. by gluing, pressing or burning). During or after the fixation, the carrier substance is removed and the substrate is immobilized on the support. Alternatively, the carrier substance is left, for this purpose the substrate is brought from the carrier substance to the surface (e.g. by an electric field with a polyacrylamide gel as carrier substance) and is thus fixed.
Auf diese Weise können Mikroarray-Chips mit Substraten in der Größenordnung von 106 und mehr in einer Serie von 1000 bis 106 Stückzahlen hergestellt werden. Beispielsweise ist ein Chip mit 104 Substrat-Punkten von 100 Mikrometer Durchmesser und einer Trennschichtdicke von ebenfalls 100 Mikrometer (lOOx 2x 100 Mikrometer)2 = 2 x 2 cm2 = 4 cm2 groß, und bei einer Scheibendicke von 100 Mikrometer gehen aus einem Block von 10 cm Länge 1000 Scheiben, also Chips, hervor. Anzahl und Abmessungen der Substrat-Punkte entsprechen der Dimension bei zur Zeit hergestellten Chips durch andereIn this way, microarray chips with substrates in the order of 10 6 and more can be produced in a series of 1000 to 10 6 pieces. For example, there is a chip with 10 4 substrate points of 100 microns in diameter and one Interface layer thickness of also 100 micrometers (100 x 2 x 100 micrometers) 2 = 2 x 2 cm 2 = 4 cm 2 , and with a slice thickness of 100 micrometers, a block of 10 cm length gives 1000 slices, i.e. chips. The number and dimensions of the substrate points correspond to the dimension in the case of chips currently being produced by others
Herstellungsverfahren.Production method.
Die herausragenden Vorteile dieser Methode sind:The outstanding advantages of this method are:
1. Die ökonomische und schnelle Herstellung von einem Vielfachen der Chipmengen, die nach den bisher üblichen Herstellungsverfahren in einer Serie produziert werden. Beispielsweise ergibt ein Faserbündel von 10 cm Länge 1000 Chips. Diese Zahl erreicht bei den Streifenblöcken eine Million Chips, da bei einer Schichtabmessung von 10 cm x 10 cm parallel 1000 Blöcke verarbeitet werden.1. The economical and rapid production of a multiple of the chip quantities that are produced in a series according to the previously usual manufacturing processes. For example, a fiber bundle of 10 cm in length gives 1000 chips. This number reaches one million chips for the stripe blocks, since 1000 blocks are processed in parallel with a layer size of 10 cm x 10 cm.
2. Die geringe Variabilität der Chips einer Serie. Dadurch, dass alle Substrate in dem Faserbündel oder Streifenblock in der Längsachse absolut homogen sind und die einzelnen Substrat-Punkte alle die gleiche Größe haben, hängt die Variabilität unter den Chips einer Serie lediglich von der Präzision des Schneidevorgangs ab. Anders als die individuelle Punkt- Auftragung bei der konventionellen Technik der Mikroarray-Chipherstellung, erfolgt der Schnitt in einem Zug durch den gesamten Block, so dass die Variabilität der individuellen Substrat-Punkte prinzipiell geringer ist.2. The low variability of the chips in a series. Because all the substrates in the fiber bundle or strip block are absolutely homogeneous in the longitudinal axis and the individual substrate points are all of the same size, the variability among the chips in a series depends only on the precision of the cutting process. In contrast to the individual point application in the conventional technique of microarray chip production, the cut is made in one go through the entire block, so that the variability of the individual substrate points is in principle less.
Die Anwendungsgebiete sind bereits jetzt als immens einzuschätzen, eine genaue Abschätzung des gesamten potenziellen Anwendungsgebiets ist kaum möglich. Angespornt durch das Humane Genomprojekt haben beispielsweise DNS-Mikrochips bereits massiven Einzug in Forschungsinstitute und in die pharmazeutische Industrie erhalten, und weitere Anwendungsbereiche eröffiien sich in rasanter Folge. Ein möglicherweise noch größerer Markt ist voraussichtlich in der klinischen Anwendung für Screening-Untersuchungen. Die bisherigen Herstellungtechniken sind relativ ineffizient, d.h. gemessen an dem finanziellen und zeitlichen Aufwand ist die Produktion gering. Dadurch ist bereits jetzt die Nachfrage größer als das Angebot, und dies wird sich in den kommenden Jahren verschärfen, vor allem nach der Fertigstellung des Humanen Genomprojekts. Wichtiger noch ist, dass aufgrund des hohen technischen Aufwandes die konventionellen Chips sehr teuer sind und damit die Chips für viele Anwendungen, die wissenschaftlich und medizinisch sinnvoll wären, finanziell unerschwinglich machen. Das hier vorgestellte Verfahren wird die Produktionskosten und damit die Preise drastisch senken können und gleichzeitig eine qualitative Verbesserung der Chips erreichen. The areas of application can already be assessed as immense, an exact assessment of the entire potential area of application is hardly possible. Spurred on by the human genome project, DNA microchips, for example, have already found their way into research institutes and the pharmaceutical industry, and further areas of application are opening up in rapid succession. A potentially even larger market is expected to be in clinical use for screening tests. The previous manufacturing techniques are relatively inefficient, i.e. measured by the financial and time expenditure, the production is low. As a result, demand is already greater than supply, and this will intensify in the coming years, especially after the completion of the human genome project. It is even more important that the conventional chips are very expensive due to the high technical complexity, making the chips financially unaffordable for many applications that would make scientific and medical sense. The process presented here will be able to drastically reduce production costs and thus prices, while at the same time achieving a qualitative improvement in the chips.

Claims

Patentansprüche claims
1. Verfahren zur Herstellung von Mikroarray-Chips, dadurch gekennzeichnet, dass1. A method for producing microarray chips, characterized in that
- ein Substrat mit einer Trägersubstanz homogen vermischt wird,a substrate is mixed homogeneously with a carrier substance,
- die Trägersubstanz mit dem Substrat in eine Form gebracht wird,the carrier substance is brought into a shape with the substrate,
- verschiedene Substrat-Trägersubstanz-Gemische zu einem Bündel zusammensetzt werden,various substrate-carrier substance mixtures are combined to form a bundle,
- anschließend das Bündel in dünne Scheiben geschnitten wird,- then the bundle is cut into thin slices,
- die Scheiben auf eine geeignete feste Unterlage gebracht werden und- The panes are placed on a suitable firm surface and
- schließlich die Trägersubstanz entfernt und dabei das Substrat auf der Unterlage immobilisiert wird.- Finally, the carrier substance is removed and the substrate is immobilized on the base.
2. Verfahren nach Anspruch 1, gekennzeichnet durch ein biologisches oder chemisches Testmaterial als Substrat.2. The method according to claim 1, characterized by a biological or chemical test material as a substrate.
3. Verfahren nach Anspruch 1 und 2, gekennzeichnet durch Nukleotidsequenzen als Substrat.3. The method according to claim 1 and 2, characterized by nucleotide sequences as a substrate.
4. Verfahren nach Anspruch 1 und 2, gekennzeichnet durch Peptidsequenzen als Substrat.4. The method according to claim 1 and 2, characterized by peptide sequences as a substrate.
5. Verfahren nach Anspruch 1, gekennzeichnet durch ein Material als Trägersubstanz, das von einem formbaren Zustand in einen festen Aggregatzustand versetzt werden kann, ohne das Substrat zu beschädigen.5. The method according to claim 1, characterized by a material as a carrier substance, which can be changed from a moldable state to a solid physical state without damaging the substrate.
6. Verfahren nach Anspruch 1 und 5, gekennzeichnet durch Paraffin als Trägersubstanz.6. The method according to claim 1 and 5, characterized by paraffin as a carrier.
7. Verfahren nach Anspruch 1 und 5, gekennzeichnet durch Polyacrylamid als Trägersubstanz.7. The method according to claim 1 and 5, characterized by polyacrylamide as a carrier.
8. Methode zur Verarbeitung und Bündelung von Substrat-Trägersubstanz-Gemischen nach Anspruch 1, dadurch gekennzeichnet, dass8. Method for processing and bundling substrate-carrier substance mixtures according to claim 1, characterized in that
- das Substrat-Trägersubstanz-Gemisch in Form von Fasern gezogen oder gegossen wird und- The substrate-carrier substance mixture is drawn or cast in the form of fibers and
- eine gewünschte Anzahl von Fasern mit unterschiedlichen Substraten zu einem geordneten Bündel zusammengesetzt werden.- A desired number of fibers with different substrates can be assembled into an ordered bundle.
9. Methode nach Anspruch 8, dadurch gekennzeichnet, dass Fasern von beliebigem Durchmesser, insbesondere auch von Durchmessern unter 100 Mikrometer, hergestellt werden. 9. The method according to claim 8, characterized in that fibers of any diameter, in particular also of diameters below 100 micrometers, are produced.
10. Methode nach Anspruch 8, gekennzeichnet durch eine sonstige Anordnung der Fasern mit verschiedenen Substraten, die geignet ist, um die Fasern im Querschnitt eindeutig zu identifizieren.10. The method according to claim 8, characterized by another arrangement of the fibers with different substrates, which is suitable to uniquely identify the fibers in cross section.
11. Methode nach Anspruch 8, dadurch gekennzeichnet, dass Fasern von beliebigem Querschnitt, insbesondere auch von quadratischem Querschnitt, hergestellt werden.11. The method according to claim 8, characterized in that fibers of any cross-section, in particular also of square cross-section, are produced.
12. Methode nach Anspruch 8, dadurch gekennzeichnet, dass Fasern mit verschiedenen Substraten in einem konzentrischen Raster (Abb. la) geordnet werden.12. The method according to claim 8, characterized in that fibers with different substrates are arranged in a concentric grid (Fig. La).
13. Methode nach Anspruch 8, dadurch gekennzeichnet, dass Fasern mit verschiedenen Substraten in einem rechteckigen Raster (Abb. lb) geordnet werden.13. The method according to claim 8, characterized in that fibers with different substrates are arranged in a rectangular grid (Fig. Lb).
14. Methode zur Verarbeitung und Bündelung von Substrat-Trägersubstanz-Gemischen nach Anspruch 1, dadurch gekennzeichnet, dass14. Method for processing and bundling substrate-carrier substance mixtures according to claim 1, characterized in that
- das Substrat-Trägersubstanz-Gemisch in Form von Schichten gebracht wird,the substrate-carrier substance mixture is brought in the form of layers,
- beliebig viele Schichten mit unterschiedlichen Substraten aufeinander geschichtet werden, die Schichten in Streifen geschnitten werden und schließlich die Streifen zu einem Bündel zusammengesetzt werden.- Any number of layers with different substrates are layered on top of each other, the layers are cut into strips and finally the strips are put together to form a bundle.
15. Methode nach Anspruch 14, gekennzeichnet durch eine beliebige Dicke der Schichten, insbesondere auch Dicken von unter 100 Mikrometer.15. The method according to claim 14, characterized by any thickness of the layers, in particular also thicknesses of less than 100 microns.
16. Methode nach Anspruch 14, dadurch gekennzeichnet, dass zwei übereinander liegende Schichten durch eine Trennschicht separiert sind (Abb. 2a).16. The method according to claim 14, characterized in that two layers lying one above the other are separated by a separating layer (Fig. 2a).
17. Methode nach Anspruch 14, dadurch gekennzeichnet, dass die zusammengesetzten Schichten mit beliebig vielen Lagen in Streifen (Abb. 2b) geschnitten werden.17. The method according to claim 14, characterized in that the composite layers with any number of layers are cut into strips (Fig. 2b).
18. Methode nach Anspruch 14, dadurch gekennzeichnet, dass geschnittenen Streifen eine beliebige Breite haben, insbesondere Breiten von unter 100 Mikrometer.18. The method according to claim 14, characterized in that cut strips have any width, in particular widths of less than 100 microns.
19. Methode nach Anspruch 14, dadurch gekennzeichnet, dass geschnittenen Streifen mit anderen Streifen an den Schnittstellen zusammengesetzt werden (Abb. 2c).19. The method according to claim 14, characterized in that cut strips are assembled with other strips at the interfaces (Fig. 2c).
20. Methode nach Anspruch 14, dadurch gekennzeichnet, dass zusammengesetzten Streifen durch eine Trennschicht separiert sind (Abb. 2c).20. The method according to claim 14, characterized in that composite strips are separated by a separating layer (Fig. 2c).
21. Methode zur Herstellung von Mikroarray-Chips aus Bündeln nach Anspruch 1, dadurch gekennzeichnet, dass das Bündel quer zur Längsachse in Scheiben geschnitten wird,21. Method for producing microarray chips from bundles according to claim 1, characterized in that the bundle is cut into slices transversely to the longitudinal axis.
- die Scheiben auf eine geeignete feste Unterlage gebracht werden,- the panes are placed on a suitable firm surface,
- die Trägersubstanz entfernt und das Substrat auf der Unterlage immobilisiert wird. - The carrier substance is removed and the substrate is immobilized on the base.
22. Methode nach Anspruch 21, gekennzeichnet durch Scheiben beliebiger Dicke, insbesondere von Dicken unter 100 Mikrometer.22. The method according to claim 21, characterized by discs of any thickness, in particular of thicknesses less than 100 microns.
23. Methode nach Anspruch 21, gekennzeichnet durch ein festes Material als Unterlage, das die Haftung des Substrates gewährleistet.23. The method according to claim 21, characterized by a solid material as a base, which ensures the adhesion of the substrate.
24. Methode nach Anspruch 21, gekennzeichnet durch unbehandeltes Glas als Material der festen Unterlage.24. The method according to claim 21, characterized by untreated glass as the material of the solid base.
25. Methode nach Anspruch 21, gekennzeichnet durch behandeltes Glas als Material der festen Unterlage.25. The method according to claim 21, characterized by treated glass as the material of the solid base.
26. Methode nach Anspruch 21, dadurch gekennzeichnet, dass die Trägersubstanz chemisch herausgelöst wird.26. The method according to claim 21, characterized in that the carrier substance is chemically dissolved out.
27. Methode nach Anspruch 21, dadurch gekennzeichnet, dass die Trägersubstanz physikalisch herausgelöst wird, z.B. Verdampfung durch Wärme-Einwirkung.27. Method according to claim 21, characterized in that the carrier substance is physically extracted, e.g. Evaporation through exposure to heat.
28. Methode nach Anspruch 21, dadurch gekennzeichnet, dass die Trägersubstanz biologisch herausgelöst wird, z.B. Verdauung durch Enzyme.28. Method according to claim 21, characterized in that the carrier substance is biologically extracted, e.g. Digestion by enzymes.
29. Methode nach Anspruch 21, dadurch gekennzeichnet, dass das Substrat nicht-kovalent auf der Unterlage immobilisiert wird.29. The method according to claim 21, characterized in that the substrate is immobilized non-covalently on the base.
30. Methode nach Anspruch 21, dadurch gekennzeichnet, dass das Substrat kovalent an der Unterlage gebunden wird.30. The method according to claim 21, characterized in that the substrate is covalently bound to the base.
31. Methode nach Anspruch 21, dadurch gekennzeichnet, daß31. The method according to claim 21, characterized in that
- das Trägermaterial nicht entfernt wird,- the carrier material is not removed,
- das Substrat an die Oberfläche der Scheibe gebracht wird,the substrate is brought to the surface of the pane,
- das oberflächliche Substrat mit dem darunter liegenden Trägermatrial an der Unterlage fixiert wird. - The superficial substrate with the underlying carrier material is fixed to the base.
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