WO2009133679A1 - Procédé de fabrication d'une électrode pour biodétecteur et procédé de fabrication d'un biodétecteur - Google Patents

Procédé de fabrication d'une électrode pour biodétecteur et procédé de fabrication d'un biodétecteur Download PDF

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WO2009133679A1
WO2009133679A1 PCT/JP2009/001881 JP2009001881W WO2009133679A1 WO 2009133679 A1 WO2009133679 A1 WO 2009133679A1 JP 2009001881 W JP2009001881 W JP 2009001881W WO 2009133679 A1 WO2009133679 A1 WO 2009133679A1
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electrode
biosensor
substrate
manufacturing
nanocarbon
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PCT/JP2009/001881
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English (en)
Japanese (ja)
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松本達
二瓶史行
成田薫
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日本電気株式会社
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • C12Q1/005—Enzyme electrodes involving specific analytes or enzymes
    • C12Q1/006—Enzyme electrodes involving specific analytes or enzymes for glucose
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon

Definitions

  • the present invention relates to a biosensor electrode manufacturing method and a biosensor manufacturing method.
  • Patent Document 1 discloses a method in which a carbon nanotube is micelleed with a mixture of a surfactant and a polymer electrolyte solution, dispersed in an aqueous solvent, and then the micelle is decomposed to form a thin film of the carbon nanotube. It is disclosed.
  • Patent Document 2 discloses that carbon nanotubes are dispersed in an optical modeling resin, and the optical modeling resin is cured by light irradiation to form an electrode.
  • Patent Document 3 discloses a method of forming a carbon nanotube film by a dry method. Specifically, a raw material gas is sprayed on the catalyst supporting surface of the substrate to grow carbon nanotubes on the catalyst supporting surface.
  • the present invention provides a method for producing a biosensor electrode and a method for producing a biosensor that can improve the performance of the biosensor.
  • a method for producing an electrode for a biosensor comprising the step of immobilizing an antibody or an enzyme on the nanocarbon layer.
  • nanocarbon is any of carbon nanotubes, carbon nanohorns, carbon nanocones, carbon nanofilaments, and fullerenes.
  • a biosensor manufacturing method including the above-described biosensor electrode manufacturing method, wherein an alignment mark is formed at a position below the nanocarbon layer of the substrate.
  • a step of mounting the biosensor electrode on a mounting substrate, and the step of mounting the biosensor electrode on the mounting substrate includes forming an alignment mark on the biosensor electrode substrate on the mounting substrate.
  • a biosensor manufacturing method can be provided in which alignment is performed using the alignment mark and the biosensor electrode is mounted on the mounting substrate.
  • the manufacturing method of the electrode for biosensors and the manufacturing method of a biosensor which can improve the performance of a biosensor are provided.
  • FIG. 1 discloses a biosensor 1 of the present embodiment.
  • the biosensor 1 includes a biosensor body 11 including a substrate 12, a working electrode 13 formed on the substrate 12, a counter electrode 14, and a reference electrode 15, and a mounting substrate 16.
  • the substrate 12 and the working electrode 13 constitute a carbon electrode for the biosensor of the present invention.
  • the substrate 12 is an insulating substrate and is a light transmissive substrate.
  • the substrate 12 is glass, plastic, or the like, and the material is not particularly limited, but glass having high strength and high light transmission is preferable.
  • light-transmitting means transmitting visible light, and it is only necessary to transmit light through the light-transmitting object to such an extent that the opposite side can be visually recognized by the naked eye.
  • the light transmittance of visible light is preferably 70% or more.
  • the working electrode 13 includes a carbon nanotube thin film (nanocarbon layer) 131 formed on the substrate 12, a layer 132 containing an antibody and an enzyme provided so as to cover the thin film 131, and a thin film 131 provided on the layer 132. And a polymer film 134 provided so as to cover the surface.
  • the thin film 131 is formed by depositing carbon nanotubes. As shown in FIG. 2, the carbon nanotubes intersect each other so as to be in direct contact with each other.
  • reference numeral 131A indicates a carbon nanotube
  • a dotted line A indicates an intersection of the carbon nanotubes 131A. Since the polymer film 134 is provided on the thin film 131, a polymer may be interposed between some of the carbon nanotubes 131A (less than 1% of all the carbon nanotubes). The carbon nanotubes 131A (99% or more of all carbon nanotubes) are in direct contact with each other.
  • the carbon nanotube 131A is in direct contact with the substrate 12, and the thin film 131 is physically fixed (adsorbed) to the substrate 12 (see dotted line B in FIG. 2). Since the polymer film 134 is provided on the thin film 131, a polymer may be interposed between some of the carbon nanotubes 131A and the substrate 12, but most of the carbon nanotubes 131A are in direct contact with the substrate 12. is doing.
  • Various types of carbon nanotubes can be used. For example, they are produced by a laser ablation method, a CoMoCAT method (a method of the University of Oklahoma, USA), a direct-injection pyrolysis synthesis method, or a high pressure carbon monoxide process method. Kinds are preferably used.
  • the laser ablation method is more preferable from the viewpoint of cost.
  • the thin film 131 is light-transmitting, and it is sufficient that the thin film 131 has such a transparency that the substrate 12 can be visually recognized through the thin film 131. Specifically, the light transmittance of visible light is preferably 70% or more.
  • Carbon nanotubes having a diameter of 0.8 to 1.3 nm and a length of several ⁇ m are preferably used from the viewpoints of quality, difficulty of peeling from the substrate 12, and the like.
  • the thickness of the thin film 131 is preferably 10 nm or more and 1 ⁇ m or less. More preferably, it is 20 nm or more and 50 nm or less. If the thickness of the thin film 131 is too thin, the electrical conductivity decreases, which is not preferable from the viewpoint of reducing the amount of antibody or enzyme immobilized. On the other hand, if it is too thick, the light transmittance is lowered, which is not preferable.
  • the layer 132 containing an antibody and an enzyme shown in FIG. 1 is obtained by fixing an antibody and an enzyme on a thin film 131.
  • the carbon nanotube thin film 131 is preliminarily treated with an immobilized protein, and then a solution containing the antibody is brought into contact therewith.
  • the antibody 132A is immobilized on the carbon nanotube 131A via the immobilized protein 132C.
  • the immobilized protein has a function for immobilizing the antibody on the carbon nanotube.
  • proteins containing electrochemically active amino acids such as tryptophan and tyrosine may be used, and streptavidin, protein A, protein G and the like are preferably used.
  • electrochemically active amino acid if the above-mentioned electrochemically active amino acid is contained in the measurement target substance, these amino acids may not be contained in the immobilized protein. Moreover, even if it is not contained in both the substance to be measured and the immobilized protein, the solution used at the time of measurement may contain an electrochemically active amino acid. These selections can be appropriately changed depending on the substance to be measured.
  • an antibody to be used it is possible to apply a wide range of antigens, such as human chorionic gonadotropin antibody and trinitrotoluene antibody, from an antigen having a large molecular weight to a small antigen.
  • an enzyme immobilization method a method using a crosslinking reaction between albumin and glutaraldehyde is preferably used because it can be produced at low cost.
  • an oxidase such as glucose oxidase, lactate oxidase, or urate oxidase is mainly used, but a dehydrogenase can also be used using a mediator or the like.
  • the layer 132 is a layer containing an antibody and an enzyme, but may be a layer having only one of an antibody and an enzyme.
  • the polymer film 134 covers the layer 132 and the thin film 131 from the upper surface side, and is provided to firmly fix the thin film 131 to the substrate 12.
  • the polymer film 134 also serves to reinforce that the antibody or the like is immobilized on the thin film 131.
  • the polymer film 134 include polyvinyl alcohol, polyacetylene, polythiophene, proteins such as albumin and gelatin, and the like. Of these, polyvinyl alcohol is preferably used because it is inexpensive and the solution can be easily adjusted.
  • the counter electrode 14 is disposed so as to face the working electrode 13 and includes, for example, platinum.
  • the reference electrode 15 includes, for example, silver. Both the counter electrode 14 and the reference electrode 15 are formed on the substrate 12.
  • the biosensor body 11 having the above configuration is mounted on a mounting substrate 16.
  • the mounting substrate 16 is an insulating substrate, for example, a flexible substrate.
  • the mounting substrate 16 is preferably made of a material excellent in water resistance, heat resistance, and chemical resistance, and plastics that can be manufactured at low cost are preferable.
  • a wiring (not shown) is formed on the mounting substrate 16, and the biosensor body 11 is connected to the wiring via a bonding wire W. Sealing agent E is provided on the bonding wire portion.
  • the sealant E is not particularly limited as long as it uses a material that waterproofs the bonding wire W and further the connection portion between the bonding wire W and the biosensor main body 11 and protects it from breakage.
  • a silicone resin is preferably used from the viewpoint of reliability.
  • the biosensor body 11 is connected to the electrochemical measurement device via the mounting substrate 16.
  • the manufacturing method of the biosensor 1 of the present embodiment includes a step of manufacturing the biosensor main body 11 and a step of mounting the biosensor main body 11 on the mounting substrate 16.
  • the manufacturing method of the biosensor body 11 includes a step of manufacturing a carbon electrode including the working electrode 13 and the substrate 12, and a step of manufacturing the counter electrode 14 and the reference electrode 15.
  • the steps of manufacturing the carbon electrode include adding a nanocarbon (carbon nanotube) to a solvent that does not contain a binder agent to create a dispersed liquid (carbon nanotube dispersion), and the above-mentioned liquid in which nanocarbon is dispersed. And a step of coating on the substrate 12 to form a nanocarbon layer (thin film 131), and a step of immobilizing an antibody or an enzyme on the nanocarbon layer.
  • the binder agent is for adhering nanocarbons to each other, and is, for example, a polymer material such as a polymer resin.
  • carbon nanotubes are prepared, and the carbon nanotubes are washed using an acid or alkali solution.
  • carbon nanotubes are dispersed in an acid or alkali solution and washed.
  • the acid solution preferably has a pH of 4 or less, and examples thereof include nitric acid and hydrochloric acid.
  • the concentration of nitric acid, hydrochloric acid, etc. in the acid solution is preferably 0.01M or more and 1M or less.
  • As an alkaline solution it is preferable that pH is 8 or more, for example, ammonia, sodium hydroxide, potassium hydroxide etc. are mentioned.
  • the concentration is preferably 0.01M or more and 3M or less.
  • the used catalyst may be slightly attached to the carbon nanotube. Such a catalyst may affect the sensitivity of the biosensor. Therefore, by providing this step, the catalyst adhering to the carbon nanotube is removed, and the sensitivity of the biosensor can be prevented from being lowered. Whether or not the catalyst has been removed can be confirmed by, for example, EDX (energy dispersive X-ray spectrometry).
  • EDX energy dispersive X-ray spectrometry
  • the carbon nanotubes are dispersed in a solvent that does not contain a binder agent for bonding the carbon nanotubes together to create a carbon nanotube dispersion.
  • a solvent an organic solvent can be used.
  • the organic solvent dichloroethane, dimethylsulfone, acetone, ether, DMF (dimethylformamide) and the like are preferable. Of these, it is preferable to use dichloroethane having good dispersibility of carbon nanotubes.
  • the solvent may be stirred with a stirrer.
  • the carbon nanotubes are efficiently dissolved in the solvent. Can be dispersed. Among these, carbon nanotubes can be more effectively dispersed in the solvent by using dichloroethane and further stirring with ultrasonic waves.
  • the carbon nanotube dispersion liquid is composed only of carbon nanotubes and a solvent, and does not contain a binder agent, a surfactant or the like.
  • a solvent in which nanocarbon is dispersed (carbon nanotube dispersion) is applied on the substrate 12 to form a carbon nanotube thin film 131.
  • a spin coating method or a dip coating method is preferably used as the coating method, but the dip coating method is preferable from the viewpoint that the thickness of the thin film 131 can be increased efficiently. Even when the spin coating method is applied, it is possible to increase the thickness of the carbon nanotube thin film 131 by increasing the number of coatings as in the case of the dip coating method.
  • a polymer film 134 is formed. Specifically, a solution containing a polymer (for example, a polyvinyl alcohol solution) is applied on the layer 132. Thereafter, the counter electrode 14 and the reference electrode 15 are formed on the substrate 12. The biosensor body 11 is manufactured through the above steps.
  • a polymer for example, a polyvinyl alcohol solution
  • an alignment mark is formed on the substrate 12 of the biosensor body 11 in advance.
  • a metal such as gold or silver may be formed in advance by sputtering or the like and patterned, or may be scratched with a glass cutter or the like.
  • the former is preferable when the size of the substrate 12 is small, and the latter is preferable when the size of the substrate 12 is large.
  • the step of forming the alignment mark M1 is performed on the substrate 12 before the electrode is formed. In the state where the electrode is formed, the alignment mark M1 is covered with the carbon nanotube thin film 131 of the working electrode 13 or the like.
  • the alignment mark M2 is also formed on the mounting substrate 16.
  • the wiring is formed on the mounting substrate 16, it is preferable to pattern the alignment mark with the same material as the wiring, but the alignment mark M2 is not limited to this.
  • the alignment mark M2 formed on the surface of the mounting substrate 16 is aligned with the alignment mark M1 formed on the substrate 12, and the biosensor body 11 is placed on the mounting substrate 16.
  • the substrate 12 is light transmissive, and the carbon nanotube thin film 131 and the polymer film 134 are also light transmissive, in this embodiment, alignment can be easily performed.
  • the mounting substrate 16 and the biosensor main body 11 are electrically connected using the bonding wire W, and the connection portion is sealed with a sealant.
  • the effect of this embodiment is demonstrated.
  • carbon nanotubes are dispersed in a polymer material such as a resin, and this polymer material is applied onto a substrate. Resistance value becomes high.
  • the thin film 131 is formed by dispersing the carbon nanotubes in a solvent that does not contain the binder agent and applying the solvent onto the substrate 12. Therefore, the carbon nanotubes in the thin film 131 are easily brought into direct contact with each other, and it is possible to prevent the resistance value between the carbon nanotubes from increasing. Thereby, the performance of the biosensor using the carbon electrode of this embodiment can be improved.
  • a catalyst is supported on a substrate to form a carbon nanotube thin film.
  • iron, cobalt, nickel or the like is generally used as a catalyst, but such a catalyst reduces the sensitivity of the biosensor. This is because the catalyst generates an interference current in current-potential measurement or constant potential measurement (for example, Fe ⁇ Fe2 ++ 2e ⁇ ).
  • a catalyst is supported on a substrate to form a carbon nanotube thin film. After forming a carbon nanotube thin film, even if an attempt is made to remove the catalyst with an acid or alkali solution, the carbon nanotube This thin film makes it difficult for an acid or alkali solution to penetrate to the substrate surface. Therefore, it is difficult to remove the catalyst.
  • the thin film 131 is formed by applying a solvent in which carbon nanotubes are dispersed on the substrate 12, and the catalyst is supported on the substrate as in the conventional case, and the carbon nanotube thin film is formed. Does not form. Therefore, the performance of the biosensor can be improved.
  • the carbon nanotubes are washed with an acid or alkali solution, and the catalyst adhered in the carbon nanotube production process is removed, so that the catalyst can be reliably removed, and the performance of the biosensor is further improved. Can be improved.
  • the polymer film 134 is formed so as to cover the thin film 131 of carbon nanotubes.
  • the polymer film 134 can prevent the thin film 131 from peeling off the substrate 12. Since the polymer film 134 is applied on the thin film 131 after the thin film 131 is formed, the polymer hardly enters between the carbon nanotubes.
  • the thin film 131 is light transmissive.
  • the substrate 12 is also light transmissive. Therefore, the position of the biosensor body 11 with respect to the mounting substrate 16 is accurately determined by using the alignment mark M1 formed on the portion below the thin film 131 of the substrate 12 and the alignment mark M2 formed on the mounting substrate 16. Can be adapted.
  • one working electrode 13, one counter electrode 14, and one reference electrode 15 are formed on one substrate 12.
  • the present invention is not limited to this, and a large-sized substrate is prepared, and the working electrode, the counter electrode, After forming many sets of reference electrodes, the biosensor body 11 may be manufactured by cutting the substrate.
  • the carbon nanotubes are washed with an acid or alkali solution to remove the catalyst, but this step may be omitted. By doing in this way, the manufacturing process of a biosensor can be simplified.
  • the carbon nanotube was used for the thin film 131 which comprises a carbon electrode
  • the carbon used for a carbon electrode should just be nanocarbon, carbon nanohorn, carbon nanocone, carbon nanofilament, It may be fullerene.
  • Example 1 A 10 ⁇ 10 ⁇ 0.7 mm glass substrate (light-transmitting substrate) was prepared, and ultrasonic cleaning was performed in a nitric acid-hydrogen peroxide solution for 5 minutes. Subsequently, a diamond-shaped mark was formed at the center of the glass substrate as an alignment mark by sputtering at two locations. The material of the alignment mark is platinum, and the size of the alignment mark is 200 ⁇ m square. Subsequently, CoMoCAT carbon nanotubes were dispersed in 1,2-dichloroethane. In dispersion, ultrasonic treatment was performed for 2 hours to prepare a solution having a carbon nanotube concentration of 10 ppm.
  • the carbon nanotubes have a diameter of 0.8 to 1.6 nm and a length of several ⁇ m.
  • the solution in which the carbon nanotubes are dispersed does not include a binder agent or a surfactant, and includes carbon nanotubes and a solvent.
  • the glass substrate was immersed in the solution and pulled up with a dip coater (pickup speed: 0.2 mm / sec) to coat the carbon nanotubes on the substrate surface. This pulling operation was performed 10 times and 20 times, and a carbon electrode of a carbon nanotube thin film having a thickness of about 10 nm was manufactured.
  • a 22.5 w / v% albumin solution containing glucose oxidase and 1 v / v% glutaraldehyde was spin-coated at 3000 rpm to immobilize the enzyme on the carbon nanotube thin film.
  • the alignment mark formed on the glass substrate was covered with the carbon nanotube thin film, it could be visually confirmed, and the carbon nanotube thin film was light-transmitting.
  • a reference electrode and a platinum counter electrode were formed on the substrate, and a glucose biosensor main body was manufactured. Thereafter, a printed board (mounting board) made of polyimide resin of 15 ⁇ 100 ⁇ 0.2 mm in which two rhombus alignment marks were formed with copper was prepared.
  • the biosensor main body was mounted on a printed circuit board using an actual microscope so that the marks of both the alignments overlapped. Subsequently, the mounting substrate and the biosensor main body were connected by wire bonding, and the connection portion and the entire wire bonding were sealed with silicone resin. Using the biosensor thus produced, glucose in the solution (glucose concentration 1 mM) was measured by the three-electrode method.
  • Example 2 A 10 ⁇ 10 ⁇ 0.7 mm glass substrate (light-transmitting substrate) was prepared, and ultrasonic cleaning was performed in a nitric acid-hydrogen peroxide solution for 5 minutes. Subsequently, a diamond-shaped mark was formed at the center of the glass substrate as an alignment mark by sputtering at two locations. The material is platinum and the size is 200 ⁇ m square. Subsequently, laser ablation carbon nanotubes were dispersed in 1,2-dichloroethane. As a dispersion method, ultrasonic treatment was performed for 2 hours. And the solution whose density
  • the carbon nanotube has a diameter of about 1.3 nm and a length of several ⁇ m.
  • the solution in which the carbon nanotubes are dispersed does not include a binder agent or a surfactant, and includes carbon nanotubes and a solvent.
  • the glass substrate was dipped in the solution and pulled up with a dip coater (pickup speed: 0.2 mm / sec) to coat the carbon nanotubes on the substrate surface. This pulling operation was performed 20 times, and a carbon electrode of a carbon nanotube thin film having a thickness of about 16 nm was manufactured.
  • the carbon electrode was immersed in a solution of 1 mg / 0.2 ml of human chorionic gonadotropin antibody (mouse immunized monoclonal antibody manufactured by Hitest) for 1 hour. Then, it is immersed in 3 mM 1,3-diaminobenzene (Aldrich Co., USA, containing phosphate buffer solution of pH 7.4 and 0.1 M sodium chloride), and 0 to 0.8 V is 2 mV. Swept 100 times at / s and applied for 5 hours at 0.65 V to immobilize the antibody described above.
  • human chorionic gonadotropin antibody mouse immunized monoclonal antibody manufactured by Hitest
  • the alignment mark formed on the glass substrate was covered with a carbon nanotube thin film, an antibody layer, and a polyvinyl alcohol layer, but it can be visually confirmed, and the carbon nanotube thin film is light-transmitting. I understand. Thereafter, a printed board (mounting board) made of polyimide resin of 15 ⁇ 100 ⁇ 0.2 mm in which two rhombus alignment marks were formed with copper was prepared.
  • the biosensor main body was mounted on a printed circuit board using an actual microscope so that the marks of both the alignments overlapped. Subsequently, the mounting substrate and the biosensor main body were connected by wire bonding, and the connection portion and the entire wire bonding were sealed with silicone resin.
  • the carbon electrode thus produced as a working electrode the chorionic gonadotropin concentration (30 nM) in the solution was measured by a square wave voltammetry method using a glass reference electrode and a platinum counter electrode. The measurement conditions are 0.1-1.2 V sweep range, 40 mV pulse potential, 4 Hz frequency, 10 mV step potential.
  • Example 3 A 10 ⁇ 10 ⁇ 0.7 mm glass substrate (light-transmitting substrate) was prepared, and ultrasonic cleaning was performed in a nitric acid-hydrogen peroxide solution for 5 minutes. Subsequently, a diamond-shaped mark was formed at the center of the glass substrate as an alignment mark by sputtering at two locations. The material of the alignment mark is platinum, and the size of the alignment mark is 200 ⁇ m square. Subsequently, CoMoCAT carbon nanotubes were placed in a 1 M hydrochloric acid solution and subjected to ultrasonic cleaning treatment for 1 hour.
  • the carbon nanotube was scooped up with the glass filter, the carbon nanotube was rinsed fully with the pure water, and hydrochloric acid was removed completely. Thereby, the catalyst adhering to the carbon nanotube was removed. Subsequently, the above-mentioned carbon nanotubes were put in 1,2-dichloroethane and sufficiently dispersed by ultrasonic treatment for 2 hours to prepare a solution having a carbon nanotube concentration of 10 ppm.
  • the carbon nanotubes have a diameter of 0.8 to 1.6 nm and a length of several ⁇ m.
  • the solution in which the carbon nanotubes are dispersed does not include a binder agent or a surfactant, and includes carbon nanotubes and a solvent.
  • the glass substrate was immersed in the solution and pulled up with a dip coater (pickup speed: 0.2 mm / sec) to coat the carbon nanotubes on the substrate surface.
  • This pulling operation was performed 10 times and 20 times, and a carbon electrode of a carbon nanotube thin film having a thickness of about 10 nm was manufactured.
  • a 22.5 w / v% albumin solution containing glucose oxidase and 1 v / v% glutaraldehyde was spin-coated at 3000 rpm to immobilize the enzyme on the carbon nanotube thin film.
  • the alignment mark formed on the glass substrate was covered with the carbon nanotube thin film, it could be visually confirmed, and the carbon nanotube thin film was light-transmitting.
  • a reference electrode and a platinum counter electrode were formed on the substrate, and a glucose biosensor main body was manufactured. Thereafter, a printed board (mounting board) made of polyimide resin of 15 ⁇ 100 ⁇ 0.2 mm in which two rhombus alignment marks were formed with copper was prepared. Next, the biosensor main body was mounted on a printed circuit board using an actual microscope so that the marks of both the alignments overlapped. Subsequently, the mounting substrate and the biosensor main body were connected by wire bonding, and the connection portion and the entire wire bonding were sealed with silicone resin. Using the biosensor thus produced, glucose in the solution (glucose concentration 1 mM) was measured by the three-electrode method.
  • Example 4 A 10 ⁇ 10 ⁇ 0.7 mm glass substrate (light-transmitting substrate) was prepared, and ultrasonic cleaning was performed in a nitric acid-hydrogen peroxide solution for 5 minutes. Subsequently, a diamond-shaped mark was formed at the center of the glass substrate as an alignment mark by sputtering at two locations. The material of the alignment mark is platinum, and the size of the alignment mark is 200 ⁇ m square. Subsequently, CoMoCAT carbon nanotubes were placed in a 0.5 M sodium hydroxide solution and subjected to ultrasonic cleaning treatment for 1 hour. And the carbon nanotube was scooped up with the glass filter, the carbon nanotube was rinsed fully with the pure water, and the sodium hydroxide solution was removed completely.
  • the catalyst adhering to the carbon nanotube was removed.
  • the above-mentioned carbon nanotubes were put in 1,2-dichloroethane and sufficiently dispersed by ultrasonic treatment for 2 hours to prepare a solution having a carbon nanotube concentration of 10 ppm.
  • the carbon nanotubes have a diameter of 0.8 to 1.6 nm and a length of several ⁇ m.
  • the solution in which the carbon nanotubes are dispersed does not include a binder agent or a surfactant, and includes carbon nanotubes and a solvent.
  • the glass substrate was immersed in the solution and pulled up with a dip coater (pickup speed: 0.2 mm / sec) to coat the carbon nanotubes on the substrate surface.
  • This pulling operation was performed 10 times and 20 times, and a carbon electrode of a carbon nanotube thin film having a thickness of about 10 nm was manufactured.
  • a 22.5 w / v% albumin solution containing glucose oxidase and 1 v / v% glutaraldehyde was spin-coated at 3000 rpm to immobilize the enzyme on the carbon nanotube thin film.
  • the alignment mark formed on the glass substrate was covered with the carbon nanotube thin film, it could be visually confirmed, and the carbon nanotube thin film was light-transmitting.
  • a reference electrode and a platinum counter electrode were formed on the substrate, and a glucose biosensor main body was manufactured.
  • a printed board (mounting board) made of polyimide resin of 15 ⁇ 100 ⁇ 0.2 mm in which two rhombus alignment marks were formed with copper was prepared.
  • the biosensor main body was mounted on a printed circuit board using an actual microscope so that the marks of both the alignments overlapped.
  • the mounting substrate and the biosensor main body were connected by wire bonding, and the connection portion and the entire wire bonding were sealed with silicone resin.
  • glucose in the solution glucose in the solution (glucose concentration 1 mM) was measured by the three-electrode method.
  • a glass substrate of 10 ⁇ 10 ⁇ 0.7 mm was prepared and subjected to ultrasonic cleaning for 5 minutes in a nitric acid-hydrogen peroxide solution. Subsequently, a diamond-shaped mark was formed at the center of the glass substrate as an alignment mark by sputtering at two locations.
  • the material of the alignment mark is platinum, and the size of the alignment mark is 200 ⁇ m square.
  • CoMoCAT carbon nanotubes were dispersed in 1,2-dichloroethane, and further dispersed by ultrasonic treatment for 2 hours.
  • carbon nanotubes were placed in a 5 w / v% Nafion (registered trademark) solution manufactured by DuPont, which is a polymer electrolyte solution, and further subjected to ultrasonic cleaning treatment for 2 hours, and Nafion (registered trademark) having a concentration of 10 ppm.
  • -A carbon nanotube solution was prepared.
  • the carbon nanotubes have a diameter of 0.8 to 1.6 nm and a length of several ⁇ m.
  • the glass substrate was immersed in the solution and pulled up by a dip coater (pickup speed: 0.2 mm / sec) to coat Nafion (registered trademark) -carbon nanotubes on the substrate surface.
  • This pulling operation was performed 10 times and 20 times, and a carbon electrode of a thin film of Nafion (registered trademark) -carbon nanotubes having a thickness of about 1 ⁇ m was manufactured.
  • a 22.5 w / v% albumin solution containing glucose oxidase and 1 v / v% glutaraldehyde was spin-coated at 3000 rpm, and the enzyme was immobilized on the Nafion (registered trademark) -carbon nanotube thin film. .
  • a reference electrode and a platinum counter electrode were formed on the substrate, and a glucose biosensor main body was manufactured.
  • a printed board (mounting board) made of polyimide resin of 15 ⁇ 100 ⁇ 0.2 mm in which two rhombus alignment marks were formed with copper was prepared.
  • the biosensor main body was mounted on a printed circuit board using an actual microscope so that the marks of both the alignments overlapped.
  • the mounting substrate and the biosensor main body were connected by wire bonding, and the connection portion and the entire wire bonding were sealed with silicone resin.
  • glucose in the solution glucose in the solution (glucose concentration 1 mM) was measured by the three-electrode method.
  • a 22.5 w / v% albumin solution containing glucose oxidase and 1 v / v% glutaraldehyde was spin-coated at 3000 rpm to immobilize the enzyme on the carbon nanotube thin film.
  • a reference electrode and a platinum counter electrode were formed on the substrate, and a glucose biosensor main body was manufactured.
  • the side surface of the substrate was aligned with the alignment mark of the polyimide resin described above, and the substrate was mounted on the polyimide resin.
  • the biosensor main body was mounted on a printed circuit board using an actual microscope so that the marks of both the alignments overlapped.
  • the mounting substrate and the biosensor main body were connected by wire bonding, and the connection portion and the entire wire bonding were sealed with silicone resin.
  • glucose in the solution glucose concentration 1 mM was measured by the three-electrode method.
  • Example 1 The results from Example 1 to Comparative Example 2 are shown in Table 1.
  • the items evaluated were as follows, and each item was evaluated on a 5-point scale.
  • (1) Base current value indicating the background current output value When the base current value exceeds 1 ⁇ A When the one-point base current value exceeds 500 nA and below 1 ⁇ A When the two-point base current value exceeds 200 nA and below 500 nA When the 3-point base current value exceeds 50 nA and is 200 nA or less 4-point base current value is 50 nA or less 5 points
  • (2) Indicates the current output value against noise when measuring 1 mM glucose or 30 nM chorionic gonadotropin Signal / noise average value Signal / noise average value is less than 1 1 point Signal / noise average value is 1 or more and less than 2 2 points Signal / noise average value is 2 or more and less than 5 3 Point When the average value of signal / noise is 5 or more and less than 20 4 points When the average value of signal / noise is 20 or more Total 5 points

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Abstract

La présente invention concerne une étape de la fabrication d'une électrode de carbone comprenant une sous-étape consistant à ajouter du nanocarbone à un solvant ne contenant pas d'agent liant de façon à préparer un liquide dispersé (liquide de dispersion de nanotubes de carbone) ; une sous-étape consistant à déposer sur un substrat (12) le liquide dans lequel a été dispersé le nanocarbone de façon à former une couche de nanocarbone ; et une sous-étape consistant en une solidification d'un anticorps ou d'un catalyseur au niveau de la couche de nanocarbone.
PCT/JP2009/001881 2008-04-30 2009-04-24 Procédé de fabrication d'une électrode pour biodétecteur et procédé de fabrication d'un biodétecteur WO2009133679A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014002999A1 (fr) * 2012-06-25 2014-01-03 合同会社バイオエンジニアリング研究所 Électrode enzymatique
JPWO2020175526A1 (fr) * 2019-02-27 2020-09-03
JP2020528950A (ja) * 2017-07-27 2020-10-01 プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ 電気伝導性防汚コーティング組成物

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004177237A (ja) * 2002-11-26 2004-06-24 Matsushita Electric Works Ltd 半導体イオンセンサ
JP2006308463A (ja) * 2005-04-28 2006-11-09 National Institute Of Advanced Industrial & Technology ナノカーボンセンサー
JP2006317360A (ja) * 2005-05-16 2006-11-24 National Institute Of Advanced Industrial & Technology 竹型カーボンナノチューブを用いたバイオセンサ
WO2008066123A1 (fr) * 2006-11-30 2008-06-05 National Institute Of Advanced Industrial Science And Technology Biocapteur utilisant un nanotube de carbone

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004177237A (ja) * 2002-11-26 2004-06-24 Matsushita Electric Works Ltd 半導体イオンセンサ
JP2006308463A (ja) * 2005-04-28 2006-11-09 National Institute Of Advanced Industrial & Technology ナノカーボンセンサー
JP2006317360A (ja) * 2005-05-16 2006-11-24 National Institute Of Advanced Industrial & Technology 竹型カーボンナノチューブを用いたバイオセンサ
WO2008066123A1 (fr) * 2006-11-30 2008-06-05 National Institute Of Advanced Industrial Science And Technology Biocapteur utilisant un nanotube de carbone

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014002999A1 (fr) * 2012-06-25 2014-01-03 合同会社バイオエンジニアリング研究所 Électrode enzymatique
CN104520700A (zh) * 2012-06-25 2015-04-15 日本生物工程研究所有限责任公司 酶电极
JPWO2014002999A1 (ja) * 2012-06-25 2016-06-02 合同会社バイオエンジニアリング研究所 酵素電極
JP2020528950A (ja) * 2017-07-27 2020-10-01 プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ 電気伝導性防汚コーティング組成物
JP7365330B2 (ja) 2017-07-27 2023-10-19 プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ 電気伝導性防汚コーティング組成物
JPWO2020175526A1 (fr) * 2019-02-27 2020-09-03
WO2020175526A1 (fr) * 2019-02-27 2020-09-03 国立研究開発法人産業技術総合研究所 Microstructure, son procédé de fabrication et procédé de détection de molécule l'utilisant
JP7291422B2 (ja) 2019-02-27 2023-06-15 国立研究開発法人産業技術総合研究所 微小構造体、その作製方法およびそれを利用した分子検出方法

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