WO2002052260A2 - Biosensor and method of producing the same - Google Patents

Biosensor and method of producing the same Download PDF

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Publication number
WO2002052260A2
WO2002052260A2 PCT/EP2001/015225 EP0115225W WO02052260A2 WO 2002052260 A2 WO2002052260 A2 WO 2002052260A2 EP 0115225 W EP0115225 W EP 0115225W WO 02052260 A2 WO02052260 A2 WO 02052260A2
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WIPO (PCT)
Prior art keywords
layer
protective layer
biosensor
plasmon
hydrogel
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PCT/EP2001/015225
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German (de)
French (fr)
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WO2002052260A3 (en
WO2002052260A9 (en
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Andreas Hofmann
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Jandratek Gmbh
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Priority to AU2002237253A priority Critical patent/AU2002237253A1/en
Publication of WO2002052260A2 publication Critical patent/WO2002052260A2/en
Publication of WO2002052260A9 publication Critical patent/WO2002052260A9/en
Publication of WO2002052260A3 publication Critical patent/WO2002052260A3/en

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    • 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
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Definitions

  • the invention relates to a biosensor with a support, a plasmon-supporting layer arranged on the support made of a metal with free electrons or a semiconductor, an intermediate layer and an external hydrogel layer with the possibility of connecting receptors for the binding of analytes and the like Property to avoid unspecific adsorption.
  • Such a biosensor is e.g. known from DE 198 17 180 C2.
  • the biosensor whose hydrogel layer consists of a polymer, for example dextran, amose or celulose and whose receptors are formed, for example, by proteins (e.g. antibodies, antigens, membrane proteins), glycoproteins, nykleotids or DNA, to which an analyte ( Binding partner) can serve to detect certain molecules (in particular biomolecules) in a sample to be examined, for which purpose an optical plasmon resonance measurement is carried out.
  • proteins e.g. antibodies, antigens, membrane proteins
  • glycoproteins e.g. antibodies, antigens, membrane proteins
  • nykleotids or DNA to which an analyte ( Binding partner) can serve to detect certain molecules (in particular biomolecules) in a sample to be examined, for which purpose an optical plasmon resonance measurement is carried out.
  • a covalently bound organic compound predominantly sulfur compound
  • the plasmon-bearing noble metal layer in particular gold layer
  • the intermediate layer made of a polymer
  • a further linker is used to bind the hydrogel (main polymer).
  • the object of the invention is to create a biosensor which is comparatively simple to manufacture and which is insensitive to disturbing contamination of the plasmon-bearing layer.
  • the intermediate layer contains a protective layer covering the plasmone-bearing layer, which is physisorbed onto the plasmone-bearing layer and to which the hydrogel layer is connected.
  • the physisorption provided according to the invention makes chemical binding in the manner of a linker to the plasmon-carrying layer superfluous.
  • the physisorbed protective layer for example a very thin silicon oxide layer, protects the plasmon-bearing layer in an excellent manner against impurities, in particular in combination with the hydrogel layer it prevents the so-called unspecific adsorption. This result is already achieved with a thin protective layer of less than 10 nm, in particular less than 5 nm, which seals the plasmon-carrying layer, without this protective layer losing the physical effect relevant for the measurements.
  • the hydrogel layer can be more easily connected to the physisorbed protective layer and that known and proven coating methods can be adopted from other areas. Furthermore, when the plasmon-bearing noble metal layer is applied by vapor deposition or sputtering onto the carrier, the protective layer can also be applied in one step in a labor-saving manner.
  • the invention also relates to a method for producing the biosensor according to the invention, in which a plasmon-carrying layer is applied to a carrier, which is then covered with a hydrogel layer with the interposition of an intermediate layer.
  • this method is characterized in that after the plasmon-carrying layer has been applied to the intermediate layer, a protective layer is physisorbed, whereupon the hydrogel layer is attached to the intermediate layer.
  • FIGS. 3 and 4 the result of optical plasmon resonance measurements (detector response curves) with a biosensor according to FIG. 1 with a physisorbed protective layer (FIG. 4) and without such a protective layer ( Figure 3) show.
  • a plasmon-carrying layer 2 is arranged on a carrier 1, for example a glass prism, which can be a noble metal layer, in particular a gold layer, a silver layer or a semiconductor layer. Between the Carrier 1 and layer 2 can be provided with an adhesion promoter (not shown). As shown in FIG. 1, there is an intermediate layer 3 on layer 2.
  • An ultra-thin silicon monoxide was applied to layer 2 by means of an adhesion promoter, for example nickel / chromium, titanium, chromium, by means of thermal evaporation, sputtering, electron beam evaporation, sol gel technology or plasma polymerization Layer (Sio layer) of less than 5 nm thickness applied.
  • the SiO is oxidized to SiC> 2 in air and forms a protective layer 4 on the plasmon-carrying layer 2.
  • a hydrogel layer 6 e.g. Dextran, amylose or cellulose.
  • This hydrogel layer 6 can be provided with receptors 7 in a known manner, e.g. Proteins (e.g. antibodies, antigens, membrane proteins), glycoproteins, nucleotides, DNA, to which only the analytes to be detected, mainly biomolecules, attach, provided that they are in a sample to be applied to the hydrogel layer 6.
  • FIG. 2 shows a biosensor modified compared to FIG. 1, but which in the same way has a carrier 1, a plasmon-carrying layer 2 and a hydrogel layer 6 with receptors 7.
  • the intermediate layer 8 has a protective layer 9, for the formation of which a polymer, for example polyacrylic acid, was applied to the surface of the layer 2 by means of plasma polymerization.
  • the hydrogel layer 6 is attached to this polymer layer 9 serving as a protective layer by means of a linker 10.

Abstract

The invention relates to a biosensor comprising a support (1) (prism of glass), a plasmon-carrying layer (2) that consists of a metal layer with free electrons, especially gold or silver, or a semiconductor layer and a hydrogel layer (6) that consists of a polymer, especially a dextran to which receptors (7) can be bound. Said biosensor is further provided with a protective layer (4) that consists of a silicon oxide or a metal oxide, or a protective layer (9) that consists of a polymer and that is physisorbed to the plasmon-carrying layer (2). The hydrogel layer (6) can be bound via a linker (5, 10), for example in the case of a protective layer (4) from silicon oxide by for example silane as the linker (5). The protective layer (4) has a thickness of less than 10 nm, preferably less than 5 nm, and effectively protects the active plasmon-carrying layer (2) from any disturbing contaminations. The physical optical effect produced by the surface plasmon resonance does not take place outside the range that can be measured so that the suitability as a biosensor is not lost.

Description

Bezeichnung: Biosensor und Verfahren zu seinerName: Biosensor and method for its
Herstellungmanufacturing
B e s c h r e i b u n gDescription
Die Erfindung bezieht sich auf einen Biosensor mit einem Träger, einer auf dem Träger angeordneten, plasmonentragenden Schicht aus einem Metall mit freien Elektronen bzw. einem Halbleiter, einer Zwischenschicht und einer außenliegenden Hydrogelschicht mit der Möglichkeit zum Anbinden von Rezeptoren für die Bindung von Analyten und der Eigenschaft, unspezifische Adsorption zu vermeiden.The invention relates to a biosensor with a support, a plasmon-supporting layer arranged on the support made of a metal with free electrons or a semiconductor, an intermediate layer and an external hydrogel layer with the possibility of connecting receptors for the binding of analytes and the like Property to avoid unspecific adsorption.
Ein derartiger Biosensor (Affinitätssensor) ist z.B. aus DE 198 17 180 C2 bekannt.Such a biosensor (affinity sensor) is e.g. known from DE 198 17 180 C2.
Der Biosensor, dessen Hydrogelschicht aus- einem Polymer, beispielsweise Dextran, Am lose oder Celulose besteht und dessen Rezeptoren beispielsweise von Proteinen (z. B. Antikörper, Antigenen, Membranproteinen) , Glykoproteinen, Nykleotiden oder DNA gebildet sind, an die sich ein Analyt (Bindungspartner) anlagern kann, dient dem Nachweis bestimmter Moleküle (insbesondere Biomoleküle) in einer zu untersuchenden Probe, wozu eine optische Plasmonenresonanzmessung vorgenommen wird. Bei den bekannten Biosensoren ist eine kovalentgebundene organische Verbindung, vorwiegend Schwefelverbindung, als Linker zwischen der plasmonentragenden Edelmetallschicht (insbesondere Goldschicht) und der Zwischenschicht aus einem Polymer vorgesehen, und ein weiterer Linker dient zum Anbinden des Hydrogels (Hauptpolymer) . Dieser Aufbau erfordert nicht nur eine vergleichsweise aufwändige Herstellung sondern hat auch den Nachteil, daß es zu einer störenden Verunreinigung der reaktiven Goldschicht durch Substanzen, z.B. von organischen Verbindungen aus der Luft, kommen kann, was wiederum eine aufwändige Reinigungsprozedur bedingt.The biosensor, whose hydrogel layer consists of a polymer, for example dextran, amose or celulose and whose receptors are formed, for example, by proteins (e.g. antibodies, antigens, membrane proteins), glycoproteins, nykleotids or DNA, to which an analyte ( Binding partner) can serve to detect certain molecules (in particular biomolecules) in a sample to be examined, for which purpose an optical plasmon resonance measurement is carried out. In the known biosensors, a covalently bound organic compound, predominantly sulfur compound, is provided as a linker between the plasmon-bearing noble metal layer (in particular gold layer) and the intermediate layer made of a polymer, and a further linker is used to bind the hydrogel (main polymer). This structure not only requires a comparatively complex production but also has the disadvantage that the reactive gold layer can be contaminated by substances, for example organic compounds from the air, which in turn requires a complex cleaning procedure.
Dementsprechend liegt der Erfindung die Aufgabe zugrunde, einen Biosensor zu schaffen, der sich vergleichsweise einfach herstellen läßt und der unanfällig gegen störende Verunreinigung der plasmonentragenden Schicht ist.Accordingly, the object of the invention is to create a biosensor which is comparatively simple to manufacture and which is insensitive to disturbing contamination of the plasmon-bearing layer.
Diese Aufgabe wird erfindungsgemäß dadurch gelöst, daß die Zwischenschicht eine die plasmonentragende Schicht abdeckende Schutzschicht enthält, die auf die plasmonentragende Schicht physisorbiert ist und mit der die Hydrogelschicht verbunden ist.This object is achieved according to the invention in that the intermediate layer contains a protective layer covering the plasmone-bearing layer, which is physisorbed onto the plasmone-bearing layer and to which the hydrogel layer is connected.
Die erfindungsgemäß vorgesehene Physisorption, also eine Bindung über elektrostatische Kräfte, macht ein chemisches Anbinden nach Art eines Linkers an die plasmonentragende Schicht überflüssig. Die physisorbierte Schutzschicht, beispielsweise eine.sehr dünne Siliziumoxidschicht, schützt die plasmonentragende Schicht in hervorragender Weise vor Verunreinigungen, insbesondere verhindert sie in Kombination mit der Hydrogelschicht die sogenannte unspezifische Adsorption. Dieses Ergebnis wird bereits mit einer die plasmonentragende Schicht versiegelnden dünnen Schutzschicht von weniger als 10 nm, ins- besonder weniger als 5 nm, erreicht, ohne daß durch diese Schutzschicht der für die Messungen relevante physikalische Effekt verloren geht. Weitere Vorteile des erfindungsgemäßen Biosensors bestehen darin, daß sich die Hydrogelschicht einfacher an die physisor- bierte Schutzschicht anbinden läßt und sich bekannte und bewährte Beschichtungs ethoden aus anderen Bereichen übernehmen lassen. Weiter läßt sich beim Aufbringen der plasmonentragenden Edelmetallschicht durch Aufdampfen oder Sputtern auf den Träger in arbeitssparender Weise in einem Schritt auch die Schutzschicht aufbringen.The physisorption provided according to the invention, that is to say binding via electrostatic forces, makes chemical binding in the manner of a linker to the plasmon-carrying layer superfluous. The physisorbed protective layer, for example a very thin silicon oxide layer, protects the plasmon-bearing layer in an excellent manner against impurities, in particular in combination with the hydrogel layer it prevents the so-called unspecific adsorption. This result is already achieved with a thin protective layer of less than 10 nm, in particular less than 5 nm, which seals the plasmon-carrying layer, without this protective layer losing the physical effect relevant for the measurements. Further advantages of the biosensor according to the invention are that the hydrogel layer can be more easily connected to the physisorbed protective layer and that known and proven coating methods can be adopted from other areas. Furthermore, when the plasmon-bearing noble metal layer is applied by vapor deposition or sputtering onto the carrier, the protective layer can also be applied in one step in a labor-saving manner.
Die Erfindung betrifft auch ein Verfahren zum Herstellen des erfindungsgemäßen Biosensors, bei dem auf einen Träger eine plasmonentragende Schicht aufgebracht wird, die dann unter Zwischenfügung einer Zwischenschicht mit einer Hydrogelschicht überdeckt wird.The invention also relates to a method for producing the biosensor according to the invention, in which a plasmon-carrying layer is applied to a carrier, which is then covered with a hydrogel layer with the interposition of an intermediate layer.
Dieses Verfahren ist erfindungsgemäß dadurch gekennzeichnet, daß nach dem Aufbringen der plasmonentragenden Schicht auf diese als Zwischenschicht eine Schutzschicht physisorbiert wird, worauf an die Zwischenschicht die Hydrogelschicht angelagert wird.According to the invention, this method is characterized in that after the plasmon-carrying layer has been applied to the intermediate layer, a protective layer is physisorbed, whereupon the hydrogel layer is attached to the intermediate layer.
Zweckmäßige Ausgestaltungen und Weiterbildungen der Erfindung ergeben sich aus den Unteransprüchen.Expedient refinements and developments of the invention result from the subclaims.
Die Erfindung wird nachfolgend anhand von zwei Beispielen und den zugehörigen schematischen Figuren 1 und 2 der beiden beispielhaften Biosensoren näher erläutert, wobei die Figuren 3 und 4 das Ergebnis optischer Plasmonenresonanzmessungen (Detektorantwortskurven) mit einem Biosensor gemäß Figur 1 mit physisorbierter Schutzschicht (Figur 4) und ohne eine solche Schutzschicht (Figur 3) zeigen.The invention is explained in more detail below with the aid of two examples and the associated schematic FIGS. 1 and 2 of the two exemplary biosensors, FIGS. 3 and 4 the result of optical plasmon resonance measurements (detector response curves) with a biosensor according to FIG. 1 with a physisorbed protective layer (FIG. 4) and without such a protective layer (Figure 3) show.
Beispiel 1:Example 1:
Auf einem Träger 1, beispielsweise ein Glasprisma, ist eine plasmonentragende Schicht 2 angeordnet, bei der es sich um eine Edelmetallschicht, insbesondere eine Goldschicht, eine Silberschicht oder eine Halbleiterschicht handeln kann. Zwischen dem Träger 1 und der Schicht 2 kann ein Haftvermittler (nicht dargestellt) vorgesehen sein. Wie in Figur 1 dargestellt, befindet sich auf der Schicht 2 eine Zwischenschicht 3. Auf die Schicht 2 wurde über einen Haftvermittler, z.B. Nickel/Chrom, Titan, Chrom, mittels thermischen Aufdampfens, Sputterns, Elektronen- strahlverdampfens, Solgeltechnik oder Plasmapolymerisation eine ultradünne Siliziummonoxid-Schicht (Sio-Schicht) von weniger als 5 nm Dicke aufgebracht. Das SiO ist an der Luft zu SiC>2 oxidiert und bildet eine Schutzschicht 4 auf der plasmonentragenden Schicht 2.A plasmon-carrying layer 2 is arranged on a carrier 1, for example a glass prism, which can be a noble metal layer, in particular a gold layer, a silver layer or a semiconductor layer. Between the Carrier 1 and layer 2 can be provided with an adhesion promoter (not shown). As shown in FIG. 1, there is an intermediate layer 3 on layer 2. An ultra-thin silicon monoxide was applied to layer 2 by means of an adhesion promoter, for example nickel / chromium, titanium, chromium, by means of thermal evaporation, sputtering, electron beam evaporation, sol gel technology or plasma polymerization Layer (Sio layer) of less than 5 nm thickness applied. The SiO is oxidized to SiC> 2 in air and forms a protective layer 4 on the plasmon-carrying layer 2.
Anschließend wurde die Oberfläche der Schutzschicht 4 silani- siert, wie es dem Fachmann bekannt und u.a. von Elam, Nygren und Stenberg im Artikel "Covalent coupling of polysaccharides to Silicon and Silicon rubber surfaces" in Journal of Biomedi- cal Materials Research, Vol. 18, 953 bis 959 (1984) beschrieben ist. Über die so gebildete Silanschicht 5 ist eine Hydrogelschicht 6, z.B. Dextran, Amylose oder Cellulose, angeschlossen. Diese Hydrogelschicht 6 kann in bekannter Weise mit Rezeptoren 7 versehen werden, z.B. Proteinen (z.B Antikörpern, Antigenen, Membranproteinen) , Glykoproteinen, Nukleotiden, DNA, an die sich nur die nachzuweisenden Analyte, vorwiegend Biomoleküle, anlagern, soweit sich diese in einer auf die Hydrogelschicht 6 aufzubringenden Probe befinden.The surface of the protective layer 4 was then silanized, as is known to the person skilled in the art and i.a. by Elam, Nygren and Stenberg in the article "Covalent coupling of polysaccharides to Silicon and Silicon rubber surfaces" in Journal of Biomedical Materials Research, Vol. 18, 953 to 959 (1984). A hydrogel layer 6, e.g. Dextran, amylose or cellulose. This hydrogel layer 6 can be provided with receptors 7 in a known manner, e.g. Proteins (e.g. antibodies, antigens, membrane proteins), glycoproteins, nucleotides, DNA, to which only the analytes to be detected, mainly biomolecules, attach, provided that they are in a sample to be applied to the hydrogel layer 6.
Zum Nachweis dafür, daß die erfindungsgemäß vorgesehene phy- sisorbierte Schutzschicht 4 die Plasmonenresonanzmessung nicht stört, wurden Vergleichsversuche für einen Biosensor ohne die Schutzschicht 4 (Figur 3) und für den Biosensor mit der Schutzschicht 4 (Figur 4) gemacht. Dabei sind die reflektierten Lichtintensitäten über den durchgestimmten Einfallswinkeln des auf die Unterseite des Trägers 1 gelenkten monochromatischen Lichts dargestellt. Für beide Fälle ist das resonanzbedingte Intensitatsminimum, das einen Hinweis auf die an den Rezeptoren 7 angelagerten Biomoleküle gibt, annähernd gleich deutlich zu erkennen. Dabei entspricht die im Vergleich zu Figur 3 in Figur 4 vorhandene Verschiebung und Verbreiterung der Kurven den theoretischen Annahmen. Exakte Meßergebnisse sind also trotz der vorhandenen Schutzschicht 4 gewährleistet. Beispiel 2 :To prove that the physisorbed protective layer 4 provided according to the invention does not interfere with the plasmon resonance measurement, comparative tests were carried out for a biosensor without the protective layer 4 (FIG. 3) and for the biosensor with the protective layer 4 (FIG. 4). The reflected light intensities are shown over the tuned angles of incidence of the monochromatic light directed onto the underside of the carrier 1. For both cases, the resonance-related minimum intensity, which gives an indication of the biomolecules attached to the receptors 7, can be recognized approximately equally clearly. The shift and widening of the curves compared to FIG. 3 in FIG. 4 correspond to the theoretical assumptions. Exact measurement results are thus guaranteed despite the protective layer 4 present. Example 2:
Figur 2 zeigt einen gegenüber Figur 1 abgewandelten Biosensor, der jedoch in gleicher Weise einen Träger 1, eine plasmonentragende Schicht 2 und eine Hydrogelschicht 6 mit Rezeptoren 7 aufweist. Abweichend weist die Zwischenschicht 8 eine Schutzschicht 9 auf, zu deren Ausbildung auf die Oberfläche der Schicht 2 mittels Plasmapolymerisation ein Polymer, beispielsweise Polyacrylsäure, aufgebracht wurde. An diese als Schutzschicht dienende Polymerschicht 9 ist die Hydrogelschicht 6 mittels eines Linkers 10 angelagert. FIG. 2 shows a biosensor modified compared to FIG. 1, but which in the same way has a carrier 1, a plasmon-carrying layer 2 and a hydrogel layer 6 with receptors 7. Deviating from this, the intermediate layer 8 has a protective layer 9, for the formation of which a polymer, for example polyacrylic acid, was applied to the surface of the layer 2 by means of plasma polymerization. The hydrogel layer 6 is attached to this polymer layer 9 serving as a protective layer by means of a linker 10.

Claims

P a t e n t a n s p r ü c h e Patent claims
1. Biosensor mit einem Träger (1) , einer auf dem Träger (1) angeordneten, plasmonentragenden Schicht (2) aus einem Metall mit freien Elektronen bzw. einem Halbleiter, einer Zwischenschicht (3, 8) und einer außenliegenden Hydrogelschicht (6) mit der Möglichkeit zum Anbinden von Rezeptoren (7) für die Bindung von Analyten und der Eigenschaft, unspezifische Adsorption zu vermeiden, dadurch gekennzeichnet, daß die Zwischenschicht (3, 8) eine die plasmonentragende Schicht (2) abdeckende Schutzschicht (4, 9) enthält, die auf die plasmonentragende' Schicht (2) physisorbiert ist und mit der die Hydrogelschicht (6) verbunden ist.1. Biosensor with a carrier (1), a plasmon-bearing layer (2) made of a metal with free electrons or a semiconductor, an intermediate layer (3, 8) and an external hydrogel layer (6) arranged on the carrier (1) the possibility of binding receptors (7) for the binding of analytes and the property of avoiding unspecific adsorption, characterized in that the intermediate layer (3, 8) contains a protective layer (4, 9) covering the plasmon-carrying layer (2), which is physisorbed on the plasmon-supporting 'layer (2) and with which the hydrogel layer (6) is connected.
2. Biosensor nach Anspruch 1, dadurch gekennzeichnet, daß die Schutzschicht (4, 9) weniger als 10 nm dick ist.2. Biosensor according to claim 1, characterized in that the protective layer (4, 9) is less than 10 nm thick.
3. Biosensor nach Anspruch 1, dadurch gekennzeichnet, daß die3. Biosensor according to claim 1, characterized in that the
Schutzschicht (4, 9) weniger als 5 nm dick ist.Protective layer (4, 9) is less than 5 nm thick.
4. Biosensor nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß die Schutzschicht (4) ein Siliziumoxid aufweist.4. Biosensor according to one of claims 1 to 3, characterized in that the protective layer (4) has a silicon oxide.
5. Biosensor nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß die Schutzschicht (4) ein Metalloxid aufweist.5. Biosensor according to one of claims 1 to 3, characterized in that the protective layer (4) has a metal oxide.
6. Biosensor nach einem der Ansprüche Ibis 3, dadurch gekennzeichnet, daß die Schutzschicht (9) ein Polymer aufweist.6. Biosensor according to one of claims 3, characterized in that the protective layer (9) comprises a polymer.
7. Biosensor nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß die Schutzschicht (9) Polyacrylsäure oder Polystyrol aufweist. 7. Biosensor according to one of claims 1 to 3, characterized in that the protective layer (9) comprises polyacrylic acid or polystyrene.
8. Biosensor nach einem der Ansprüche 1 bis 7, dadurch gekennzeichnet, daß die Hydrogelschicht (6) über einen Linker (5, 10) mit der Schutzschicht (4, 9) verbunden ist.8. Biosensor according to one of claims 1 to 7, characterized in that the hydrogel layer (6) via a linker (5, 10) with the protective layer (4, 9) is connected.
9. Biosensor nach einem der Ansprüche 1 bis 7, insbesondere nach Anspruch 4, dadurch gekennzeichnet, daß die Hydrogelschicht (6) über eine Silanschicht (5) als Linkerschicht mit der Schutzschicht (4) verbunden ist.9. Biosensor according to one of claims 1 to 7, in particular according to claim 4, characterized in that the hydrogel layer (6) via a silane layer (5) as a linker layer with the protective layer (4) is connected.
10. Verfahren zum Herstellen eines Biosensors gemäß den Ansprüchen 1 bis 9, bei dem auf einen Träger (1) eine plasmonentragende Schicht (2) aufgebracht wird, die dann unter Zwischenfügung einer Zwischenschicht (3, 8) mit einer Hydrogelschicht (6) überdeckt wird, dadurch gekennzeichnet, daß nach dem Aufbringen der plasmonentragenden Schicht (2) auf diese als Zwischenschicht (3, 8) eine Schutzschicht (4, 9) physisorbiert wird, worauf an die Zwischenschicht die Hydrogelschicht - (6) angelagert wird.10. A method for producing a biosensor according to claims 1 to 9, in which a plasmon-carrying layer (2) is applied to a carrier (1), which is then covered with a hydrogel layer (6) with the interposition of an intermediate layer (3, 8) , characterized in that after the plasmon-carrying layer (2) has been applied to the latter as an intermediate layer (3, 8), a protective layer (4, 9) is physisorbed, whereupon the hydrogel layer - (6) is attached to the intermediate layer.
11. Verfahren nach Anspruch 10, dadurch gekennzeichnet, daß die11. The method according to claim 10, characterized in that the
Schutzschicht durch thermisches Aufdampfen aufgebracht wird.Protective layer is applied by thermal vapor deposition.
12. Verfahren nach Anspruch 10, dadurch gekennzeichnet, daß die Schutzschicht durch Sputtern aufgebracht wird.12. The method according to claim 10, characterized in that the protective layer is applied by sputtering.
13. Verfahren nach Anspruch 10, dadurch gekennzeichnet, daß die13. The method according to claim 10, characterized in that the
Schutzschicht durch Elektronenstrahlverdampfen aufgebracht wird..Protective layer is applied by electron beam evaporation ..
14. Verfahren nach Anspruch iθ, dadurch gekennzeichnet, daß die Schutzschicht durch Solgeltechnik aufgebracht wird.14. The method according to claim iθ, characterized in that the protective layer is applied by solgel technology.
15. Verfahren nach Anspruch 10, dadurch gekennzeichnet, daß die15. The method according to claim 10, characterized in that the
Schutzschicht durch Plasmapolymerisation aufgebracht wird.Protective layer is applied by plasma polymerization.
16. Verfahren nach einem der Ansprüche 10 bis 15, dadurch gekennzeichnet, daß die Hydrogelschicht (6) mittels eines Linkers (5, 10) an die Schutzschicht (4, 9) angelagert wird. 16. The method according to any one of claims 10 to 15, characterized in that the hydrogel layer (6) is attached to the protective layer (4, 9) by means of a linker (5, 10).
PCT/EP2001/015225 2000-12-22 2001-12-21 Biosensor and method of producing the same WO2002052260A2 (en)

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