JP2008175697A - Method of manufacturing sensor, sensor, and detector - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method of manufacturing a sensor that has an electrode having a conductive layer showing sufficient wettability (adhesiveness) to a receptor for recognizing a liquid sample and a detecting object in the liquid sample, and can detect the detecting object accurately at high sensitivity, a sensor manufactured by the manufacturing method of the sensor, and a detector of high performance having the sensor. <P>SOLUTION: The method of manufacturing sensor 1 has a first step of forming the conductive layer by sticking metal powder to an organic binder component on a substrate 2, and a second step of performing at least one of treatment of decomposing the organic binder component in the conductive layer and treatment of introducing a hydrophilic group on the conductive layer. Counter electrodes 61-63 and reference electrodes 71-73 are constituted by the conductive layer having undergone this treatment. The method also has a third step of forming reaction layers 41-43 reacting with the detecting object on the conductive layer having undergone this treatment. Working electrodes 21-23 are constituted by the reaction layers and the conductive layer. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a method for manufacturing a sensor for detecting a biomolecule, a sensor, and a detection apparatus.

Biosensors using biological materials such as enzymes, DNA, and antibodies have been widely studied as means for selectively detecting many biomolecules such as blood glucose, cholesterol, urea, and vitamins.
In particular, biosensors that electrochemically detect products generated or consumed by the reaction of a detection target and an oxidase are actively researched, such as blood glucose level sensors and urine sugar level sensors. Has been developed as.

  Biosensors using such an oxidase include (1) a sensor that detects a decrease in oxygen concentration associated with an enzyme reaction with an oxygen sensor (oxygen electrode), and (2) hydrogen peroxide that is produced along with the enzyme reaction. That is electrolyzed on a hydrogen peroxide electrode, and the current generated at that time is detected. (3) The reaction is performed by reacting the complex of the redox molecule and enzyme and the measurement object. A reduction type mediator generated along with this is converted into an oxidation type mediator on an electrode, and a current generated at that time is detected. In addition, as an intermediate between (2) and (3), (4) the hydrogen peroxide produced by the enzyme reaction is reduced by horseradish peroxidase (HRP) electrically coupled to the mediator, Some have been reported to detect the current generated at that time.

By the way, specifically, such a biosensor is provided in contact with a reaction layer containing a biological substance and a reaction layer, and a measurement electrode that detects a movement of electric charges caused by a biological reaction as an electric current ( Working electrode), and a counter electrode and a compensation electrode (reference electrode) juxtaposed with the working electrode. And the measurement target contained in a liquid sample is detected by making a liquid sample contact a reaction layer, a counter electrode, and a reference electrode (for example, refer patent document 1).
Each electrode of such a biosensor is composed of a conductive layer formed in a predetermined pattern on a substrate. The working electrode is manufactured by supplying a liquid material containing a biological substance on the conductive layer to form a reaction layer.

The conductive layer is formed by various film forming methods, and is preferably formed by applying a conductive paste containing conductive material particles and an organic binder component onto a substrate and then drying. By using such a liquid phase process, the manufacturing process can be greatly simplified as compared to using a vapor phase process such as vapor deposition.
Here, in the biosensor based on the electrochemical detection principle, the detection target is detected based on the movement of electrons generated between the detection target and each electrode. For this reason, the surface of each electrode needs to have sufficient affinity (wetability) with respect to a sample.

However, an electrode containing an organic binder component is generally hydrophobic on the surface. On the other hand, many detection objects have hydrophilicity. For this reason, if the electrode surface is hydrophobic, the sample (detection target) cannot sufficiently adhere to the electrode, and it becomes difficult to detect the detection target. For this reason, there exists a problem that the detection accuracy and detection sensitivity of a detection target object are low.
In particular, when preparing a working electrode, a liquid material containing a biological substance is supplied onto the conductive layer to form a reaction layer. If the adhesion between the conductive layer and the liquid material is low, a reaction is performed. A layer cannot be formed.

JP-A-5-203608

  An object of the present invention is to provide an electrode having a conductive layer exhibiting sufficient wettability (adhesion) for a liquid sample and a receptor for recognizing the detection object in the liquid sample. Another object of the present invention is to provide a sensor manufacturing method capable of easily manufacturing a sensor capable of being detected with high sensitivity, a sensor manufactured by such a sensor manufacturing method, and a high-performance detection device including such a sensor.

Such an object is achieved by the present invention described below.
The method for producing a sensor of the present invention includes a first step of forming a conductive layer containing conductive material particles and an organic binder component on a substrate,
A second step of hydrophilizing the surface of the conductive layer by performing at least one of a process of decomposing the organic binder component in the conductive layer and a process of introducing a hydrophilic group onto the conductive layer. It is characterized by having.
Thereby, it is possible to easily manufacture a sensor that includes an electrode having a conductive layer exhibiting sufficient wettability (adhesion) with respect to a liquid sample and can detect a detection target with high accuracy and high sensitivity.

The sensor manufacturing method of the present invention preferably further includes a third step of forming a reaction layer that reacts with the detection target in the sample on the conductive layer subjected to the second step.
This facilitates a sensor that has an electrode having a conductive layer exhibiting sufficient wettability (adhesion) for a receptor that recognizes a detection target and can detect the detection target with high accuracy and high sensitivity. Can be manufactured.
In the sensor manufacturing method of the present invention, it is preferable that the reaction layer includes an oxidoreductase that performs an electrochemical reaction with the detection target.
In this case, the action and effect of the present invention are more remarkably exhibited.

In the sensor manufacturing method of the present invention, in the first step, a liquid material containing particles of the conductive material and the organic binder component in a solvent is supplied onto the base material to form a liquid film. It is preferable to form the conductive layer by removing the solvent from the liquid film.
This makes it possible to easily form electrodes and wiring in a short time without using a costly and laborious process such as photolithography.

In the sensor manufacturing method of the present invention, the liquid material is preferably supplied by a droplet discharge method.
Thereby, the electrode and wiring of a fine pattern can be formed more easily. In addition, the thickness and width of the electrode and the wiring can be easily controlled by controlling the number of droplets to be discharged.
In the sensor manufacturing method of the present invention, it is preferable that the treatment for decomposing the organic binder component irradiates the conductive layer with ultraviolet light.
Thereby, despite the simple dry process, the surface of the conductive layer can be hydrophilized in a short time.

In the sensor manufacturing method of the present invention, the wavelength of the ultraviolet light is preferably 200 to 300 nm.
Thereby, the organic binder component existing near the surface of the conductive layer can be efficiently decomposed while preventing excessive deterioration of the conductive layer and the mask used for selective irradiation with ultraviolet light. As a result, the surface of the conductive layer can be efficiently hydrophilized.

In the sensor manufacturing method of the present invention, the organic binder component preferably contains a polyacrylate resin or an epoxy resin.
These organic binder components are efficiently oxidized by ozone generated when irradiated with ultraviolet light in an oxygen-existing atmosphere to generate hydroxyl groups. Therefore, the conductive layer containing such an organic binder component is efficiently hydrophilized by the action of this hydroxyl group.
In the method for producing a sensor of the present invention, it is preferable that the treatment for decomposing the organic binder component is to bring an enzyme that decomposes fat into contact with the conductive layer.
Thereby, the surface of a conductive layer can be easily hydrophilized.

In the method for producing a sensor of the present invention, the enzyme that degrades fat is preferably lipase.
Thereby, lipase is contained in a large amount in the living body and is easily available. In addition, since lipase has high resistance to various organic solvents, when the treatment liquid containing the enzyme and the organic solvent is applied to the conductive layer, the enzyme is deactivated, and the decomposition action of the organic binder component by the enzyme is reduced. Can be prevented.

In the method for producing a sensor of the present invention, the organic binder component preferably includes a resin containing an ester bond.
The enzyme that degrades fat mainly contributes to the degradation of the ester bond, and therefore can efficiently decompose the resin containing the ester bond. For this reason, the surface of the conductive layer can be hydrophilized in a relatively short time.

In the method for producing a sensor of the present invention, a surface layer containing a reaction amplifying agent having a property that the decomposability by ultraviolet light irradiation is larger than that of the organic binder component is formed on the conductive layer, and then the surface layer It is preferable to introduce a hydrophilic group onto the surface layer by irradiating with UV light.
Accordingly, the surface of the conductive layer can be reliably hydrophilized in a short time even with ultraviolet light having a relatively long wavelength and relatively small light energy. In addition, by using ultraviolet light having a long wavelength, generation of ozone accompanying irradiation of ultraviolet light in an oxygen-existing atmosphere can be suppressed, so that an environmental load due to ozone can be reduced.
In the sensor manufacturing method of the present invention, the reaction amplification agent preferably contains a nitrobenzyl derivative.
Since the reaction amplifying agent containing a nitrobenzyl derivative has high photosensitivity, even if the intensity (illuminance) of ultraviolet light is small, it can be reliably processed in a short time.

In the sensor manufacturing method of the present invention, the wavelength of the ultraviolet light is preferably 300 to 400 nm.
Thereby, the photodegradable protecting group in the reaction amplifying agent can be reliably removed while preventing excessive deterioration of the conductive layer and the mask used for selectively irradiating with ultraviolet light. As a result, the surface of the conductive layer can be efficiently hydrophilized.

In the method for producing a sensor of the present invention, in the second step, a hydrophilic layer is introduced onto the conductive layer by forming a surface layer containing a polypeptide chain or a modified product thereof on the conductive layer. It is preferable.
Since the polypeptide chain or a modified product thereof has a high affinity for a biological substance, it is possible to prevent a decrease in activity of a detection target or a receptor in an adjacent reaction layer. For this reason, a sensor excellent in detection sensitivity can be obtained.

In the method for producing a sensor of the present invention, the polypeptide chain preferably contains bovine serum albumin or a modified product thereof.
Since bovine serum albumin and a modified product thereof are particularly firmly adhered to the conductive layer, the surface of the conductive layer can be reliably hydrophilized. In addition, when bovine serum albumin or a modified product thereof is contained in a reaction layer that is formed on a conductive layer and contains a receptor that interacts with a detection target, the surface layer is a conductive layer. Excellent adhesion to both reaction layers. Thereby, the electron mobility between the reaction layer and the conductive layer can be increased, and the sensitivity of the sensor can be increased.

In the sensor manufacturing method of the present invention, in the second step, it is preferable to introduce a hydrophilic group onto the conductive layer by forming a surface layer containing a surfactant on the conductive layer.
Thereby, the surface of the conductive layer can be reliably hydrophilized regardless of the state of the conductive layer. Therefore, the range of selection of the type of organic binder component can be expanded.

The sensor of the present invention is manufactured by the sensor manufacturing method of the present invention.
Thereby, the sensor which can detect a detection target object with high precision and high sensitivity is obtained.
The sensor of the present invention comprises a base material,
A counter electrode provided with a conductive layer provided on the substrate and containing particles of a conductive material and an organic binder component;
A working electrode comprising a conductive layer containing conductive material particles and an organic binder component provided on the substrate, and a reaction layer provided on the conductive layer and reacting with an object to be detected in a sample. And
At least one of the conductive layers is subjected to at least one of a treatment for decomposing the organic binder component contained and a treatment for introducing a hydrophilic group on the surface.
As a result, an electrode having a conductive layer exhibiting sufficient wettability (adhesiveness) for a liquid sample and a receptor that recognizes the detection object in the liquid sample is provided, and the detection object is accurately and highly sensitive. A sensor that can be detected is obtained.
The detection device of the present invention includes the sensor of the present invention.
Thereby, a high-performance detection device is obtained.

Hereinafter, a sensor manufacturing method, a sensor, and a detection device of the present invention will be described in detail based on preferred embodiments shown in the accompanying drawings.
FIG. 1 is a schematic diagram (perspective view) showing a configuration of a detection device of the present invention provided with the sensor of the present invention, FIG. 2 is a plan view schematically showing the sensor of the present invention, and FIG. FIG. 4 is a sectional view taken along line AA of the sensor shown in FIG. 2, FIG. 5 is a partially enlarged view of the sectional view shown in FIG. , 7 are longitudinal sectional views for explaining a method of manufacturing the sensor shown in FIG. In the following description, the front side of the paper in FIGS. 2 and 3 is referred to as “up”, and the back side of the paper is referred to as “down”. Moreover, the upper side in FIGS. 4-7 is called "upper", and the lower side is called "lower".

The sensor of the present invention measures the amount of a target (detection target) contained in a liquid sample based on a current value taken out from an electrode.
A measurement apparatus (detection apparatus) 100 shown in FIG. 1 includes a sensor 1, an arithmetic unit 102 that includes a processing circuit 101 that analyzes a current value obtained by the sensor 1, a connector 103 that mounts the sensor 1, and a processing circuit 101. And a wiring 104 for connecting the connector 103 to the connector 103.

The sensor 1 shown in FIGS. 2 and 3 includes a detection unit 11 configured on a substrate 2 by working electrodes (electrodes) 21, 22, 23, counter electrodes 61, 62, 63, and reference electrodes 71, 72, 73, respectively. , 12 and 13 are provided.
Each of these electrodes is independently electrically connected to the processing circuit 101 via the wiring 3, the connector 103, and the wiring 104. The sensor 1 is detachable at the connector 103.
Moreover, the detection parts 11, 12, and 13 are each isolated via the partition 6, as shown in FIG.

As shown in FIG. 4, such a sensor 1 supplies a liquid sample 50 to a sample supply space 5 defined by a substrate 2 and a partition wall 6, whereby each working electrode 21, 22, 23 described later is provided. The reaction layer provided above contacts the liquid sample 50. Then, when the target in the liquid sample 50 reacts with the reaction layer, current can be taken out from the working electrodes 21, 22, and 23. Thereby, the amount of the target in the liquid sample 50 can be measured (detected) based on the change in the current value.
Here, examples of the liquid sample include body fluids such as blood, urine, sweat, lymph, cerebrospinal fluid, bile, and saliva, and processed fluids obtained by performing various treatments on these body fluids.

In addition, the target contained in the liquid sample emits electrons (e ⁻ ) by reacting with a receptor described later.
Examples of such targets include sugars such as glucose, simple proteins, proteins such as glycoproteins, alcohols, cholesterol, steroid hormones, bile acids, steroids such as bile alcohol, vitamins and hormones. , Lactic acid, bilirubin, uric acid, creatinine and the like.

Hereinafter, the configuration of each part of the sensor 1 will be described in detail. Although the sensor 1 shown in FIG. 1 has a large number of detection units, the structure of the sensor 1 will be described below with the detection unit 11, the detection unit 12, and the detection unit 13 as representatives.
The substrate 2 supports each part constituting the sensor 1 and insulates the above-described electrodes and wiring 3 from each other.

  Examples of the constituent material of the substrate 2 include various resin materials such as polyethylene, polypropylene, polystyrene, polyethylene terephthalate (PET), polyethylene naphthalate (PES), polyimide (PI), various glass materials such as quartz glass, alumina, Various ceramic materials such as zirconia can be mentioned, and one or more of these can be used in combination.

As shown in FIG. 4, a plurality (three in the following description) of working electrodes 21, 22, and 23 are provided on the substrate 2.
The working electrodes 21, 22, and 23 include reaction layers 41, 42, and 43, which will be described later, on the conductive layers 201, 202, and 203, respectively.
Among these, the conductive layers 201, 202, and 203 are each configured by binding particles of conductive material with an organic binder component.

  Examples of conductive material particles include particles composed of gold, silver, copper, platinum or alloys containing these, metal oxide particles such as ITO, carbon-based particles such as graphite, and conductive phthalocyanine pigments. Organic pigment particles such as Of these, the conductive material preferably contains gold. Since gold is particularly excellent in oxidation resistance and chemical resistance, by using particles of a conductive material containing gold, a working electrode showing excellent durability can be obtained for most samples. In addition, since gold can be easily bonded to a thiol group in particular, the surface of the conductive layer containing gold can be easily chemically modified using this property.

  On the other hand, as an organic binder component, phenol resin, epoxy resin, acrylic resin, polyester resin, vinyl ester resin, diallyl phthalate resin, oligoester acrylate resin, xylene resin, bismaleimide triazine resin, Examples thereof include various thermosetting resins such as furan resins, urea resins, polyurethane resins, melamine resins, and silicone resins, polyether resins, and the above resin structures containing a fluorocarbon chain.

  The content of the conductive material particles in the conductive layers 201, 202, and 203 is preferably 30 to 99 wt%, and more preferably 50 to 98 wt%. Thereby, the particles of the conductive material and the particles and the substrate 2 can be reliably bonded while securing the conductivity in the conductive layers 201, 202, and 203 to some extent. As a result, conductive layers 201, 202, and 203 that are securely fixed to the substrate 2 and have higher conductivity are obtained.

As shown in FIG. 4, each of the reaction layers 41, 42, and 43 is provided so that at least a part (the upper surface in the present embodiment) is exposed to the sample supply space 5. The liquid sample 50 can be brought into contact with the reaction layers 41, 42, and 43 by supplying the liquid sample 50 to the sample supply space 5.
As shown in FIG. 5, the reaction layers 41, 42, and 43 contain receptors 411, 421, and 431. Here, when the receptors 411, 421, and 431 contain an enzyme, these receptors 411, 421, and 431 react with the targets 51, 52, and 53 in the liquid sample 50, respectively, as described above ( Electrons are emitted by an oxidation reaction.

  Any of these receptors 411, 421, and 431 may react with the same type of target, but in this embodiment, they react with different types of targets 51, 52, and 53. With such a configuration, the amount of each target 51, 52, 53 can be collectively measured with a single measurement operation for the liquid sample 50 including different types of targets 51, 52, 53 simultaneously. A multi-sensor is obtained.

Examples of such receptors 411, 421, and 431 include enzymes, polypeptides such as antibodies, oligopeptides, DNA, nucleic acids such as oligonucleotides, and the like.
For example, when the receptors 411, 421, and 431 contain antibodies, the receptors 411, 421, and 431 adsorb target antigens, and specifically recognize the antigens, such as enzymes. By adsorbing a secondary antibody having a marker that promotes electron generation, electrons can be released as in the case of the aforementioned enzyme.
Among these, it is preferable that each of the receptors 411, 421, and 431 contains an enzyme. The enzyme has a high discrimination ability of the targets 51, 52, and 53, and can realize a sensor with high detection sensitivity. In addition, a sensor using an enzyme as a receptor is easy to realize simple handling.

  As the enzyme, for example, glucose oxidase (GOx), ascorbate oxidase (ASOx), lactate oxidase (LOx), uricase (uric acid oxidase) (UOx), depending on the type of target 51, 52, 53 to be measured. Galactose oxidase, pyruvate oxidase, D- or L-amino acid oxidase, amine oxidase, cholesterol oxidase, glucose oxidase, glycerol oxidase, xanthase, choline oxidase, glutamate oxidase, oxidases such as horseradish peroxidase, alcohol dehydrogenase, glutamic acid Dehydrogenase, cholesterol rudehydrogenase, aldehyde dehydrogenase, glucose Hydrogenase, fructose dehydrogenase, sorbitol dehydrogenase, it is possible to use various oxidoreductase such as dehydrogenases, such as glycerol dehydrogenase. When the receptors 411, 421, and 431 contain an oxidoreductase, the action and effect of the present invention are more remarkably exhibited.

  In the present embodiment, as shown in FIG. 5, each of the reaction layers 41, 42, 43 includes receptors 411, 421, 431 in a matrix (base material) composed of polymers 413, 423, 433. It is contained (impregnated). By configuring the reaction layers 41, 42, and 43 as described above, the receptors 411, 421, and 431 are securely held, and in each reaction layer 41, 42, and 43, the targets 51, 52, and 53 and the receptors 411, 431, Reaction with 421 and 431 can be caused more reliably.

  The polymers 413, 423, and 433 constituting the matrix are not particularly limited. For example, natural polymers (natural resins) such as bio-related polymers (animal-derived polymers) and plant-derived polymers, and synthetic polymers are used. Polymers (synthetic resins) or modified products thereof can be used, and one or more of these can be used in combination. Among them, as the polymers 413, 423, and 433 constituting the matrix, those having a bio-derived polymer or a modified product thereof as a main component are preferable. By using these, it is possible to suitably prevent the receptors 411, 421, and 431 from being easily denatured and deactivated.

Examples of such biologically relevant polymers include carbohydrates, proteins (especially albumin, globulin, myoglobin), nucleic acids (DNA, RNA) and the like.
Examples of the modified product include those subjected to treatment for breaking the hydrophobic bond, hydrogen bond, and ionic bond of the biological polymer. Examples of such treatment include heat treatment, pressure treatment, pH adjustment treatment, treatment with a denaturing agent, and the like.
The matrix preferably has a cross-linked structure. Thereby, the receivers 411, 421, and 431 can be firmly held (supported) on the matrix. Moreover, it contributes to the improvement of the mechanical strength of each reaction layer 41,42,43.

As the cross-linking agent that forms a cross-linked structure in the matrix, for example, when a polymer having a peptide as a main component is used, for example, glutaraldehyde, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, trinitromethane Water-soluble titanium oxide (titanium lactate; [(OH) ₂ Ti (C ₃ H ₅ O ₂ ) ₂ ], titanium triethanolamate; [(C ₃ H ₇ O) ₂ Ti (C ₆ H ₁₄ O ₃ N ) ₂ ]), water-soluble zirconium oxide (zirconyl acetate; [ZrO (OCOCH ₃ ) ₂ ]) and the like, and one or more of these can be used in combination.

  The content of the receptors 411, 421, 431 in each reaction layer 41, 42, 43 is preferably 5 to 50 wt%, more preferably about 40 to 50 wt% with respect to the matrix serving as the base material. preferable. Thereby, when the targets 51, 52, and 53 are included in the liquid sample 50, a sufficient reaction can be generated between the receptors 411, 421, and 431 and the targets 51, 52, and 53, and the electrons Can be efficiently released.

  Each reaction layer 41, 42, 43 preferably includes mediators 412, 422, 432 that mediate the emitted electrons to the conductive layers 201, 202, 203. The mediators 412, 422, and 432 generally generate an oxidation reaction at a lower potential than the targets 51, 52, and 53. For this reason, by using the mediators 412, 422, and 432 as an electron transfer medium, the electrons from the reaction layers 41, 42, and 43 are efficiently transferred, and the current is detected from the detection units 11, 12, and 13 with higher sensitivity. It can be taken out.

  Examples of the mediators 412, 422, and 432 include potassium ferricyanide, ferrocene or ferrocene derivatives, nickelocene or nickelocene derivatives, pyridine or pyridine derivatives, quinone or quinone derivatives (eg, p-benzoquinone, pyrroloquinoline quinone, etc.), flavin adenine dinucleotide ( Flavin derivatives such as FAD), nicotinamide adenine dinucleotide (NAD), nicotinamide derivatives such as nicotinamide adenine dinucleotide phosphate (NADP), phenazine methosulfate, 2,6-dichlorophenolindophenol, hexacyanoiron (III) Acid salts, octacyanotungsten ions, porphyrin derivatives, phthalocyanine derivatives, etc., and one or more of these In combination it can be used.

  When the reaction layers 41, 42, and 43 include the mediators 412, 422, and 432, the content thereof is preferably about 1 to 50 wt%, and preferably about 10 to 30 wt% with respect to the base material matrix. More preferred. By setting the content of the mediators 412, 422, and 432 in the above range, the electrons emitted from the reaction layers 41, 42, and 43 can be reliably and rapidly moved.

On the substrate 2, counter electrodes 61, 62, 63 are provided so as to be exposed to the sample supply space 5.
The counter electrodes 61, 62, 63 are electrodes for applying a voltage between the working electrodes 21, 22, 23, respectively. With the liquid sample 50 supplied to the sample supply space 5, a voltage is applied between the working electrodes 21, 22, 23 and the counter electrodes 61, 62, 63 so that the working electrodes 21, 22, 23 side is at a high potential. When applied, the electrons released by the reaction between the targets 51, 52, 53 and the receptors 411, 421, 431 can be reliably moved to the working electrodes 21, 22, 23 side.
Examples of the constituent material of the counter electrodes 61, 62, and 63 include the same materials as those of the conductive layers 201, 202, and 203 described above.

  The voltage applied between the working electrodes 21, 22, 23 and the counter electrodes 61, 62, 63 is preferably 0.6V or less, and more preferably about 0.4 to 0.5V. . By setting the value of the applied voltage within the above range, the electrons emitted by the reaction between the targets 51, 52, 53 and the receptors 411, 421, 431 can be moved more reliably. Moreover, when the biological sample is contained in the liquid sample 50 and the reaction layers 41, 42, and 43, the denaturation and deactivation can be reliably prevented (blocked).

Reference electrodes 71, 72, and 73 are provided on the substrate 2 so as to be exposed to the sample supply space 5.
The reference electrodes 71, 72, and 73 are electrodes that apply voltages to the counter electrodes 61, 62, and 63, respectively. In a state where the liquid sample 50 is supplied to the sample supply space 5, a voltage is applied between the reference electrodes 71, 72, 73 and the counter electrodes 61, 62, 63. Then, by comparing the current value flowing between these electrodes with the current value flowing between the aforementioned working electrodes 21, 22, 23 and the counter electrodes 61, 62, 63, the targets 51, 52, 53 and The current value generated by the reaction with the receptors 411, 421, and 431 can be measured with higher accuracy.

Each of the reference electrodes 71, 72, 73 is also configured by binding particles of conductive material with an organic binder component.
Examples of the conductive material particles constituting the reference electrodes 71, 72, and 73 include metal particles made of silver-silver chloride, mercury-mercury sulfate, or the like.
On the other hand, examples of the organic binder component include the same organic binder components used in the conductive layers 201, 202, and 203 described above.
Note that the voltages applied between the reference electrodes 71, 72, 73 and the counter electrodes 61, 62, 63 are also applied between the working electrodes 21, 22, 23 and the counter electrodes 61, 62, 63, respectively. It is preferable that the voltage is approximately the same as the voltage to be applied.
As described above, the conductive layers 201, 202, 203, the counter electrodes 61, 62, 63, the reference electrodes 71, 72, 73, and the wiring 3 are each formed by a liquid phase process. Thereby, these electrodes and wiring can be formed easily. As a result, the manufacturing process of the sensor can be greatly simplified, and the cost of the sensor can be reduced.

  By the way, as described above, the conductive layers 201, 202, 203, the counter electrodes 61, 62, 63, and the reference electrodes 71, 72, 73 are each formed by binding particles of a conductive material with an organic binder component. Therefore, as they are, the surfaces of these electrodes also show hydrophobicity due to the influence of the hydrophobicity of the organic binder component. In this case, the adhesion between the surfaces of the conductive layers 201, 202, and 203 and the reaction layers 41, 42, and 43 is lowered, and the liquid sample is applied to the surfaces of the counter electrodes 61, 62, and 63 and the reference electrodes 71, 72, and 73. 50 wettability (adhesion) decreases.

  In contrast, in the present embodiment, since the surfaces of these electrodes are made hydrophilic, the reaction layers 41, 42, and 43 are formed on the surfaces of the conductive layers 201, 202, and 203 with good adhesion, respectively. ing. Further, the liquid sample 50 exhibits good wettability (adhesion) with respect to the surfaces of the counter electrodes 61, 62, 63 and the reference electrodes 71, 72, 73, respectively. Therefore, the electrons generated by the reaction between the targets 51, 52, 53 and the acceptors 411, 421, 431 can be reliably transferred from the reaction layers 41, 42, 43 to the conductive layers 201, 202, 203. it can. Further, the functions of the counter electrodes 61, 62, 63 and the reference electrodes 71, 72, 73 can be reliably obtained, and the current values generated in the reaction layers 41, 42, 43 can be measured with high accuracy and high sensitivity. .

In addition, although each size in planar view of the detection parts 11, 12, and 13 is not specifically limited, respectively, it is preferable that it is about 0.1-5 mm, and it is more preferable that it is about 0.2-3 mm.
Moreover, each size in the planar view of the working electrodes 21, 22, and 23 is not particularly limited, but is preferably about 0.02 to 2 mm, and more preferably about 0.05 to 1 mm.

The partition wall 6 is provided so as to isolate the detection units 11, 12 and 13 from each other. The partition wall 6 forms a sample supply space 5 for storing the liquid sample 50 in the vicinity of each of the detection units 11, 12, and 13.
The constituent material of the partition 6 is not specifically limited, For example, the material similar to the constituent material of the above-mentioned board | substrate 2 can be used.

Moreover, it is preferable that the upper surface of the partition wall 6 has liquid repellency. As a result, the liquid sample 50 supplied to the upper surface of the partition wall 6 can easily move to the sample supply space 5, and the liquid sample 50 can be easily introduced into the sample supply space 5.
Furthermore, contamination (mutual contamination) of the liquid sample 50 can be prevented between the adjacent sample supply spaces 5. Thereby, the amount of the target can be measured with higher accuracy.

Next, the operation (usage method) of the sensor 1 will be described.
In the following, as the receptors 411, 421, and 431 included in the reaction layers 41, 42, and 43, ascorbate oxidase (receptor 411), lactate oxidase (receptor 421), and glucose oxidase (receptor 431), respectively. And ascorbic acid (target 51), lactate (target 52), and glucose (target 53) are used as examples of targets 51, 52, and 53 contained in the liquid sample 50, respectively.
First, the liquid sample 50 is supplied into the sample supply space 5. Thereby, at least a part of the components contained in the liquid sample 50 diffuses into the reaction layers 41, 42, 43.
The targets 51, 52, and 53 in the liquid sample 50 react with the receptors 411, 421, and 431 to emit electrons.

Hereinafter, a specific example of the reaction between such a receptor and a target will be described.
Ascorbate oxidase of receptor 411 selectively oxidizes ascorbate of target 51. As a result of this oxidation reaction, electrons are released and ascorbic acid is changed to dehydroascorbic acid.

The lactate oxidase of the receptor 421 selectively oxidizes the lactate of the target 52. By this oxidation reaction, electrons are released and lactate is changed to pyruvic acid.
Furthermore, the glucose oxidase of the receptor 431 selectively oxidizes the glucose of the target 53. By this oxidation reaction, electrons are released and glucose is changed to gluconic acid.

  The electrons emitted from each reaction layer 41, 42, 43 reach the conductive layers 201, 202, 203. At this time, in this sensor 1, the reaction layers 41, 42, 43 are formed on the surfaces of the conductive layers 201, 202, 203 with good adhesion, so that electrons emitted from the reaction layers 41, 42, 43 are emitted. However, it efficiently reaches the conductive layers 201, 202, and 203 side. And in the working electrodes 21, 22, and 23, the current value according to the number of electrons reaching per unit time is measured with high accuracy and high sensitivity.

Further, by analyzing the measured current value by the processing circuit 101, the amount of the targets 51, 52, and 53 in the liquid sample 50 can be calculated.
At this time, if a voltage is applied between these electrodes so that the working electrodes 21, 22, and 23 are at a positive potential and the counter electrodes 61, 62, and 63 are at a negative potential, It is made smoothly.

  Prior to the measurement of the current value, a voltage is applied between the reference electrodes 71, 72, 73 and the counter electrodes 61, 62, 63, respectively. As described above, the surfaces of the counter electrodes 61, 62, 63 and the reference electrodes 71, 72, 73 are hydrophilized, and the liquid sample 50 exhibits good wettability with respect to the surfaces of these electrodes. The function of the electrode can be reliably obtained, and the current value can be measured with higher accuracy and higher sensitivity.

Here, the amount of the targets 51, 52, 53 in the liquid sample 50, the number of electrons emitted by the reaction of the targets 51, 52, 53 and the receptors 411, 421, 431, the working electrodes 21, 22, The number of electrons that reach 23 per unit time and the current value measured from the working electrodes 21, 22, and 23 have a correlation with each other.
Therefore, by measuring the current value taken out from the working electrodes 21, 22, 23, the amount of the targets 51, 52, 53 contained in the liquid sample 50 can be indirectly measured.

  When the media sample 412, 422, 432 is included in the liquid sample 50, electrons emitted from the reaction layers 41, 42, 43 move through the mediators 412, 422, 432, respectively. For example, if the mediators 412, 422, 432 are potassium ferricyanide, they are reduced to potassium ferrocyanide by receiving electrons. Then, when this potassium ferrocyanide comes into contact with the conductive layers 201, 202, 203, it is oxidized again to potassium ferricyanide, and electrons move to the conductive layers 201, 202, 203. The mediators 412, 422, 432 can mediate electrons in this way. Thereby, the current value can be measured with higher accuracy.

Next, a manufacturing method of the sensor 1 (a manufacturing method of the sensor of the present invention) will be described by taking the case of manufacturing the sensor shown in FIG. 4 as an example.
[1] First, as shown in FIG. 6A, a substrate 2 is prepared.
Then, as shown in FIG. 6B, conductive layers 201, 202, 203, conductive layers 601, 602, 603, conductive layers 701, 702, 703, and wiring (not shown) are formed on the substrate 2. (First step).
These electrodes and wirings are formed by applying (supplying) a liquid material containing conductive material particles and an organic binder component onto the substrate 2 to form a liquid film, and then applying the liquid film to the liquid film as necessary. It forms by performing post-processing (for example, heating, irradiation of infrared rays, provision of ultrasonic waves, etc.).

Here, the coating method includes, for example, a spin coating method, a casting method, a micro gravure coating method, a gravure coating method, a bar coating method, a roll coating method, a wire bar coating method, a dip coating method, a spray coating method, and a screen printing method. , A flexographic printing method, an offset printing method, an ink jet method, a micro dispenser method, a micro contact printing method, and the like can be used alone or in combination. Of these, the printing method is preferably used. Accordingly, these electrodes and wirings can be easily formed without using a pattern forming method such as a photolithography method that requires a large number of steps.
Examples of the printing method include a screen printing method and a droplet discharge method (inkjet method). The droplet discharge method is preferable. According to the droplet discharge method, it is possible to more easily form electrodes and wirings with fine patterns. In addition, the thickness and width of the electrode and the wiring can be easily controlled by controlling the number of droplets to be discharged.

Hereinafter, a method of forming these electrodes and wirings by a droplet discharge method will be described.
First, particles of an electrically conductive material and an organic binder component constituting electrodes and wirings and a dispersion medium in which these are dispersed are mixed to prepare a dispersion. In addition to these, the dispersion may contain additives such as a dispersant as required. Further, part or all of the organic binder component may be dissolved in the dispersion medium.
The average particle diameter of the conductive material particles is preferably about 0.01 to 20 μm, and more preferably about 0.1 to 10 μm. Thereby, the particle | grains of an electroconductive material become what can form a fine pattern while being excellent in the dispersibility to a solvent.

Next, a dispersion liquid is supplied on the substrate 2 by a droplet discharge method to form a liquid film having the shape of each electrode and wiring.
Next, the dispersion medium is removed from the liquid film. Thereby, the electrode and wiring comprised with electroconductive material powder and the organic binder component can be formed. According to such a method, electrodes and wirings can be easily formed in a short time without using a costly and laborious process such as photolithography.
In addition, natural drying and forced drying are mentioned as a method of removing a dispersion medium from a liquid film. Moreover, as forced drying, the method of spraying gas on a liquid film, the method of decompressing the circumference | surroundings of a liquid film, the method by heat processing, etc. are mentioned.

[2] Next, as shown in FIG. 6C, the partition wall 6 is formed so as to surround each of the detection units 11, 12, and 13.
The partition wall 6 is formed, for example, by sticking a plate-like member having an opening corresponding to the region of the detection portions 11, 12, and 13 onto the substrate 2 using a bonding member such as an adhesive. Can do. Thereby, the sample supply space 5 can be formed in the vicinity of the detection units 11, 12, and 13.

Note that the joining member may be omitted, and the plate-like member and the substrate 2 may be fused to join them.
In addition, a mask is provided so as to cover the detection units 11, 12, and 13, and a film is formed on the substrate 2 in the gap between the detection units 11, 12, and 13 using various film forming methods. Can also be used.

[3] Next, surface treatment is performed on the conductive layers 201, 202, 203, the conductive layers 601, 602, 603 and the conductive layers 701, 702, 703 to make the surfaces hydrophilic (second step).
In the method for producing a sensor of the present invention, when hydrophilizing the surfaces of these conductive layers, (A) a treatment for decomposing the organic binder component in the conductive layer, and (B) a hydrophilic group on the conductive layer. At least one of the introduction processes is performed, whereby each electrode is formed. Hereinafter, each process will be described sequentially.

(A) The process which decomposes | disassembles the organic binder component in a conductive layer As a process which decomposes | disassembles the organic binder component in a conductive layer, (A-1) The process which irradiates an ultraviolet light to a conductive layer, (A-2) For example, the conductive layer may be contacted with an enzyme that decomposes fat.

(A-1) Treatment of irradiating the conductive layer with ultraviolet light First, a mask is provided so that the conductive layers 201, 202, and 203, the conductive layers 601, 602, and 603, and the conductive layers 701, 702, and 703 are exposed.
These conductive layers are irradiated with ultraviolet rays from above the mask in an oxygen-existing atmosphere. When ultraviolet rays are irradiated in an oxygen-existing atmosphere, oxygen in the atmosphere changes to ozone. Then, this ozone oxidizes the organic binder component existing near the surface of the conductive layer to generate a hydroxyl group. The generated hydroxyl groups hydrophilize the surfaces of these conductive layers in a short time despite a simple dry process.

The wavelength of ultraviolet rays used for this surface treatment is preferably about 200 to 300 nm, and more preferably about 220 to 280 nm. Thereby, the organic binder component existing in the vicinity of the surface of the conductive layer can be efficiently decomposed while preventing excessive deterioration of the conductive layer and the mask. As a result, the surface of the conductive layer can be efficiently hydrophilized.
Moreover, it is preferable that the irradiation time of an ultraviolet-ray is about 30 seconds-10 minutes.

  Such treatment with ultraviolet light is preferably performed when the organic binder component contained in the conductive layer contains at least one of polyacrylates and epoxies. These organic binder components are efficiently oxidized by ozone to generate hydroxyl groups. Therefore, the conductive layer containing such an organic binder component is efficiently hydrophilized by the action of this hydroxyl group.

(A-2) The process which contacts the enzyme which decomposes | disassembles fat in a conductive layer The enzyme which decomposes | disassembles a fat is mentioned to the enzyme used for this process. By the action of such an enzyme, the organic binder component in the conductive layer is decomposed, and thereby the surface of the conductive layer is hydrophilized. According to this method, the surface of the conductive layer can be easily hydrophilized.
This enzyme is brought into contact with the conductive layer by, for example, dissolving it in a solvent and coating this solution (treatment liquid) on the conductive layer.

As an enzyme that degrades fat, for example, an enzyme derived from the pancreas is preferable, and a lipase is more preferable. Lipase is contained in a large amount in the living body and is easily available. In addition, since lipase has high resistance to various organic solvents, when the treatment liquid containing the enzyme and the organic solvent is applied to the conductive layer, the enzyme is deactivated, and the decomposition action of the organic binder component by the enzyme is reduced. Can be prevented.
In addition, it is preferable to perform the process using such an enzyme, when the organic binder component contained in a conductive layer contains resin containing an ester bond. The enzyme that degrades fat mainly contributes to the degradation of the ester bond, and therefore can efficiently decompose the resin containing the ester bond. For this reason, the surface of the conductive layer can be hydrophilized in a relatively short time.

Examples of the resin containing an ester bond include polyacrylate resin, glycidyl ester epoxy resin, and polyester resin.
Moreover, it is preferable to use a buffer solution as the solvent for dissolving the enzyme. Thereby, the fluctuation | variation of pH of a dispersion medium can be suppressed and it can prevent reliably that a enzyme will denature | denaturate and deactivate.

  Examples of such a buffer include triethanolamine hydrochloric acid-sodium hydroxide buffer, veronal (sodium 5,5-diethylbarbiturate) -hydrochloric acid buffer, tris-hydrochloric acid buffer, glycylglycine- Sodium hydroxide buffer, 2-amino-2-methyl-1,3-propanediol-hydrochloric acid buffer, diethanolamine-hydrochloric acid buffer, boric acid buffer, sodium borate-hydrochloric acid buffer, glycine-sodium hydroxide buffer Solution, sodium carbonate-sodium bicarbonate buffer, sodium borate-sodium hydroxide buffer, sodium bicarbonate-sodium hydroxide buffer, potassium phosphate-disodium phosphate buffer, disodium phosphate-sodium hydroxide Buffer, potassium chloride-sodium hydroxide buffer, briton-robinson buffer, G Various buffer A buffer solution (liquid containing a buffering agent) and the like.

The enzyme is preferably dissolved in a liquid (treatment liquid) obtained by adding an organic solvent to the buffer solution or other aqueous solvent, and the treatment is performed using this treatment liquid. Thereby, the wettability (adhesion) between the surface of each conductive layer and the treatment liquid is improved by the action of the organic solvent. As a result, the treatment liquid can be uniformly brought into contact with the surface of each conductive layer, and the organic binder component contained in the conductive layer can be reliably decomposed.
In this case, examples of the organic solvent include dimethylformamide, dimethyl sulfoxide, 1,4-dioxane, dimethoxyethane, and the like, and one or more of them can be used in combination.

  The mixing ratio of the organic solvent in this treatment liquid is about 10 to 25 vol% when the organic solvent is dimethylformamide, 20 to 45 vol% when it is dimethyl sulfoxide, and 10 to 25 vol when it is 1,4-dioxane. %, And in the case of dimethoxyethane, it is preferably 7.5 to 25 vol%. Thereby, the wettability of the treatment liquid with respect to the surface of the electrode can be improved while inhibiting the enzyme from being denatured and deactivated by the organic solvent.

Further, the concentration of the enzyme in the treatment liquid is preferably about 0.1 to 10 wt%, and more preferably about 1 to 5 wt%. By setting the enzyme concentration in the treatment liquid within the above range, the surface of the conductive layer can be uniformly hydrophilicized without unevenness. Even if the enzyme concentration is increased beyond the above range, an effect commensurate with it cannot be expected.
The method for applying the treatment liquid is not particularly limited, and the various application methods described above can be used.

Next, the treatment liquid is supplied to the sample supply space 5 in which the conductive layers 201, 202, 203, conductive layers 601, 602, 603, and conductive layers 701, 702, 703 are formed, and left for a certain period of time. Wash and dry.
As a result, the organic binder component present in the vicinity of the surface of the electrode is decomposed by the enzyme, and a hydrophilic group (for example, a carboxyl group) is generated at the terminal. The surface of these electrodes is hydrophilized by this hydrophilic group.

The temperature of the treatment liquid while it is left here varies depending on the type of enzyme, but is preferably about 15 to 45 ° C, more preferably about 20 to 40 ° C. By setting the temperature of the treatment liquid within the above range, the enzyme is particularly activated and the organic binder component can be decomposed more efficiently.
In addition, the time for which the treatment liquid is left is preferably about 0.5 to 24 hours, and more preferably about 2 to 8 hours.

  In this treatment method, since the treatment is performed using an enzyme that is a biological substance, the treatment liquid is formed on the surfaces of the conductive layers 201, 202, and 203 even when the treatment liquid remains on the surface of the electrode. Biological substances contained in the reaction layers 41, 42, 43, counter electrodes 61, 62, 63 and reference electrodes 71, 72, 73 formed from the conductive layers 601, 602, 603 and the conductive layers 701, 702, 703 It is possible to reliably prevent the biological material contained in the liquid sample 50 in contact with the surface of the material from being denatured and deactivated.

(B) Treatment for introducing a hydrophilic group onto the conductive layer As a treatment for introducing a hydrophilic group onto the conductive layer, for example, (B-1) forming a surface layer containing a reaction amplifying agent on the conductive layer, Treatment for irradiating this surface layer with ultraviolet light, (B-2) Treatment for forming a surface layer containing a polypeptide chain or a modified product thereof on the conductive layer, (B-3) Containing a surfactant on the conductive layer Examples include a treatment for forming a surface layer.

(B-1) Forming a surface layer containing a reaction amplifying agent on the conductive layer and irradiating the surface layer with ultraviolet light First, the conductive layers 201, 202, 203, conductive layers 601, 602, 603, conductive layers On the surfaces 701, 702, and 703, a surface layer containing a reaction amplifying agent having a property that the decomposability by irradiation with ultraviolet light is larger than that of the organic binder component contained in the conductive layer is formed.

  This reaction amplifying agent is a compound having a photodegradable protecting group, and when irradiated with ultraviolet light, the photodegradable protecting group is rapidly detached as shown in FIG. A hydrophilic group is generated at the end of the portion to which the degradable protecting group was bonded. The reaction amplification agent is supported by bringing the reaction amplification agent into contact with the conductive layer. In this case, since the reaction amplifying agent is localized on the surface of the conductive layer, the ultraviolet rays irradiated to the conductive layer can be effectively contributed to the hydrophilization of the surface of the conductive layer.

The surface layer containing the reaction amplifying agent is preferably composed of a so-called self-assembled film formed of a compound having a binding functional group capable of binding to the conductive layer. Thereby, the surface layer containing the reaction amplification agent can be formed by a simple process.
When forming a self-assembled film, a reaction amplifying agent having a photodegradable protective group and a binding functional group capable of binding to a conductive layer is used.

  Examples of the photodegradable protecting group include those having a nitrobenzyl derivative skeleton, dimethoxybenzoin group, 2-nitropiperonyloxycarbonyl (NPOC) group, 2-nitroveratryloxycarbonyl (NVOC) group, α-methyl-2 -Nitropiperonyloxycarbonyl (MeNPOC) group, α-methyl-2-nitroveratryloxycarbonyl (MeNVOC) group, 2,6-dinitrobenzyloxycarbonyl (DNBOC) group, α-methyl-2,6-dinitro Benzyloxycarbonyl (MeDNBOC) group, 1- (2-nitrophenyl) ethyloxycarbonyl (NPEOC) group, 1-methyl-1- (2-nitrophenyl) ethyloxycarbonyl (MeNPEOC) group, 9-anthracenylmethyl An oxycarbonyl (ANMOC) group, 1- Renylmethyloxycarbonyl (PYMOC) group, 3'-methoxybenzoinyloxycarbonyl (MBOC) group, 3 ', 5'-dimethoxybenzoyloxycarbonyl (DMBOC) group, 7-nitroindolinyloxycarbonyl (NIOC) group 5,7-dinitroindolinyloxycarbonyl (DNIOC) group, 2-anthraquinonylmethyloxycarbonyl (AQMOC) group, α, α-dimethyl-3,5-dimethoxybenzyloxycarbonyl group, 5-bromo-7- A nitroindolinyloxycarbonyl (BNIOC) group and the like can be mentioned, and those having a nitrobenzyl derivative skeleton, specifically, those represented by the following chemical formula (1) are preferably used. Since the reaction amplifying agent containing a nitrobenzyl derivative has high photosensitivity, even if the intensity (illuminance) of ultraviolet light is small, it can be reliably processed in a short time.

［式中、Ｒ^１、Ｒ^２は、それぞれ、独立して水素原子、アルキル基またはフッ化アルキル基を表す。］

[Wherein, R ¹ and R ² each independently represents a hydrogen atom, an alkyl group, or a fluorinated alkyl group. ]

On the other hand, the binding functional group bonded to the conductive layer is appropriately selected according to the constituent material of the conductive layer. Specifically, for example, when the constituent material of the conductive layer is gold (Au), silver (Ag), copper (Cu), platinum (Pt), etc., a thiol group, a sulfide group, a disulfide group, a thiophene group, Examples include thiosulfonate groups, carboxyl groups, pyridinium groups, and the like. When the constituent material of the conductive layer is a metal oxide (such as ITO), thiol groups, halogen groups, alkoxysilyl groups, silyl halide groups, etc. Can be mentioned.
Examples of the reaction amplifying agent having these photodegradable protective groups and binding functional groups include those represented by the following chemical formulas (2) to (4).

［式中、Ｒ^３、Ｒ^４は、それぞれ、独立して水素原子、アルキル基またはフッ化アルキル基を表す。］

[Wherein, R ³ and R ⁴ each independently represents a hydrogen atom, an alkyl group or a fluorinated alkyl group. ]

［式中、Ｒ^５、Ｒ^６は、それぞれ、独立して水素原子、アルキル基またはフッ化アルキル基を表す。］

[Wherein, R ⁵ and R ⁶ each independently represents a hydrogen atom, an alkyl group, or a fluorinated alkyl group. ]

［式中、Ｒ^７、Ｒ^８は、それぞれ、独立して水素原子、アルキル基またはフッ化アルキル基を表す。］

[Wherein, R ⁷ and R ⁸ each independently represents a hydrogen atom, an alkyl group, or a fluorinated alkyl group. ]

Formation of such a self-assembled film containing a reaction amplification agent is performed as follows.
First, a reaction liquid is dissolved in a solvent to prepare a treatment liquid.
Examples of the solvent include benzene, toluene, dimethylnaphthalene, hexane, ethyl acetate, ethyl acetate, methanol, chloroform, dichloromethane, carbon tetrachloride, tetrahydrofuran, dimethyl sulfoxide, ionic solvents, etc., and these can be used alone or in a mixed solution. Can be used as
The concentration of the reaction amplification agent in the treatment liquid is preferably about 0.1 to 50 wt%, more preferably about 0.5 to 10 wt%.

Next, these treatment liquids are respectively supplied to the sample supply space 5 in which the conductive layers 201, 202, 203, the conductive layers 601, 602, 603, and the conductive layers 701, 702, 703 are formed. Wash and dry. Thereby, a binding functional group is bonded to the upper surface of each conductive layer, and a surface layer containing a reaction amplifying agent is obtained.
In this case, the temperature of the treatment liquid is preferably about 3 to 40 ° C., more preferably about 10 to 25 ° C.
Furthermore, the time for which the treatment liquid is left is preferably about 0.5 to 24 hours, and more preferably about 2 to 8 hours.

In the above description, the case where the photodegradable protective group and the binding functional group are simultaneously introduced onto the conductive layer has been described. However, these may be introduced individually.
For example, as shown in FIG. 7B, a self-assembled film containing a binding functional group is first formed on the conductive layer, and then a photodegradable protective group is introduced into the self-assembled film. It may be.
In such a method, first, as shown in FIG. 7B, a self-assembled film containing a binding functional group is formed on the conductive layer. This self-assembled film can be formed, for example, by supplying a compound such as an alkanethiol having a functional group such as an amino group on the conductive layer when the constituent material of the conductive layer is gold. it can. The compound can be supplied by, for example, applying a treatment liquid obtained by dissolving the compound in the above-described solvent on the conductive layer.
Next, as shown in FIG. 7B, a compound containing a photodegradable protective group (reaction amplification agent) is supplied onto the self-assembled film, and the photodegradable protective group is introduced into the self-assembled film. To do. Examples of the compound (reaction amplification agent) containing such a photodegradable protecting group include those represented by the following chemical formula (5).

［式中、Ｒ^９、Ｒ^１０は、それぞれ、独立して水素原子、アルキル基またはフッ化アルキル基を表す。］

[Wherein, R ⁹ and R ¹⁰ each independently represents a hydrogen atom, an alkyl group or a fluorinated alkyl group. ]

In this way, a self-assembled film before irradiation with ultraviolet light as shown in FIG. 7A can be formed.
The compound containing the photodegradable protecting group can be supplied by applying a treatment liquid obtained by dissolving this compound in the above-described solvent onto the self-assembled film. In this case, dimethyl sulfoxide is particularly preferably used as the solvent, and it is preferable that trimethylamine is contained in the solvent.

Next, a mask is provided so as to cover a region excluding a region where the conductive layers 201, 202, and 203, the conductive layers 601, 602, and 603, and the conductive layers 701, 702, and 703 are formed.
These conductive layers are irradiated with ultraviolet light from above the mask. As a result, the photodegradable protective group in the reaction amplifying agent supported on the surface of each conductive layer is eliminated (see FIG. 7A), and a hydrophilic group is generated at the terminal after the elimination. By this hydrophilic group, the surface of each conductive layer is hydrophilized.

The wavelength of ultraviolet rays used for this surface treatment is preferably about 300 to 400 nm, and more preferably about 320 to 380 nm. Thereby, the photodegradable protecting group in the reaction amplifying agent can be surely removed while preventing excessive deterioration of the conductive layer and the mask. As a result, the surface of the conductive layer can be efficiently hydrophilized.
In addition, it is preferable that the irradiation time of an ultraviolet-ray is about 30 seconds-10 minutes.

  In addition, since the reaction amplifying agent is more decomposable by ultraviolet light irradiation than the organic binder component, the surface of the conductive layer can be reliably secured in a short time even with ultraviolet light having a relatively long wavelength and relatively low light energy. Can be hydrophilized. Furthermore, by using ultraviolet light having a long wavelength, generation of ozone accompanying ultraviolet light irradiation in an oxygen-existing atmosphere can be suppressed, so that the burden on the environment due to ozone can be reduced.

  Further, in the process of decomposing the organic binder component contained in the conductive layer with ultraviolet light, the decomposability is slightly different depending on the type of the organic binder component, so that materials that can be suitably used as the organic binder component are limited to some extent. By using the reaction amplifying agent, the surface of the conductive layer can be reliably made hydrophilic regardless of the type of the organic binder component. Thereby, the range of selection of the kind of organic binder component can be expanded.

(B-2) Treatment for forming a surface layer containing a polypeptide chain or a modified product thereof on a conductive layer Polypeptide chains or modified products thereof used in this treatment include proteins, modified products thereof, oligopeptides, etc. Can be mentioned.
Here, examples of proteins include albumin such as bovine serum albumin (BSA), globulin, myoglobin, and the like, and two or more of these may be used in combination. Of these, bovine serum albumin or a modified product thereof is particularly preferably used. Since bovine serum albumin and a modified product thereof are particularly firmly adhered to the conductive layer, the surface of the conductive layer can be reliably hydrophilized. When bovine serum albumin or a modified product thereof is contained in the reaction layers 41, 42, 43, the surface layer corresponds to both the conductive layers 201, 202, 203 and the reaction layers 41, 42, 43. Excellent adhesion. Thereby, the electron mobility between the reaction layers 41, 42, and 43 and the conductive layers 201, 202, and 203 can be increased, and the sensitivity of the sensor 1 can be increased.

Examples of the denatured product of proteins include those subjected to a treatment that breaks hydrophobic bonds, hydrogen bonds, or ionic bonds in the proteins. Examples of such treatment include heat treatment, pressure treatment, pH adjustment treatment, treatment with a denaturing agent, and the like.
Such a polypeptide chain or a modified product thereof is dissolved in a solvent and applied as a treatment liquid on the conductive layer. Thereby, a surface layer containing a polypeptide chain or a modified product thereof is formed on the conductive layer.

  Here, the polypeptide chain in the surface layer or a modified product thereof spontaneously changes its polymer structure in accordance with the surface state of the conductive layer. The surface of each conductive layer described above exhibits hydrophobicity based on the organic binder component. Depending on this property, the polymer structure of the polypeptide chain or a modified product thereof may increase the interaction with the surface of each conductive layer. To change. Therefore, the formed interface between the surface layer and the conductive layer exhibits excellent adhesion.

On the other hand, a hydrophilic group (for example, an amino acid site) is exposed on the surface of the surface layer (surface opposite to the conductive layer). Thereby, the surface of the surface layer is hydrophilized.
In addition, since the polypeptide chain or a modified product thereof has a high affinity for a biological substance, it is possible to prevent the activity of the detection target and the receptor in the adjacent reaction layer from decreasing. For this reason, the sensor 1 excellent in detection sensitivity can be obtained.

A buffer solution is preferably used as a solvent for dissolving the polypeptide chain or a modified product thereof. Thereby, the fluctuation | variation of pH is suppressed, the three-dimensional structure of polypeptide changes, and it can suppress that the hydrophobicity and hydrophilicity of the surface layer containing a polypeptide chain | strand change.
As the buffer solution, the various buffer solutions described above can be used.
Moreover, it is preferable that the content rate of the polypeptide chain or its modified | denatured substance in a process liquid is about 1-5 wt%. Thereby, a surface layer containing a polypeptide chain or a modified product thereof at a necessary and sufficient density can be formed on the conductive layer. As a result, a highly hydrophilic electrode can be obtained.

Moreover, the various coating methods mentioned above are used for the method of apply | coating a process liquid.
In this case, the temperature of the treatment liquid is preferably about 15 to 45 ° C, more preferably about 20 to 40 ° C. As a result, unintended denaturation of the polypeptide chain or a modified product thereof can be prevented, and a hydrophilic group can be activated on the conductive layer to form a more hydrophilic surface layer.

In addition, after obtaining the surface layer by applying the treatment liquid on the conductive layer, a substance that promotes the polymerization of the polypeptide chain may be added before the surface layer is dried. Thereby, the polypeptide chain or a modified product thereof is networked. This networking improves the mechanical properties of the surface layer, thereby providing a surface layer that can reliably prevent peeling and the like.
Examples of the substance that accelerates the polymerization of the polypeptide chain include glutaraldehyde-containing phosphate buffer (glutaraldehyde content 2 wt%).
The surface layer thus formed on the conductive layer preferably has an average thickness of about 0.01 to 1 μm, and more preferably about 0.05 to 0.5 μm. By setting the thickness of the surface layer within the above range, the hydrophilicity of the surface layer is surely expressed regardless of the surface state of the conductive layer.

(B-3) Treatment for forming a surface layer containing a surfactant on the conductive layer The surfactant used in this treatment is dissolved in a solvent and applied as a treatment liquid on the conductive layer. Thereby, a surface layer containing a surfactant is formed on the conductive layer.
As for the surfactant in the surface layer, the hydrophobic part is adsorbed to the conductive layer by the intermolecular force (Van der Waals force). On the other hand, the hydrophilic part of the surfactant is exposed on the surface of the surface layer (the surface on the side opposite to the conductive layer), whereby the surface of the surface layer is hydrophilized. In this process, a surface layer containing a surfactant is formed on the conductive layer, thereby making the surface of the conductive layer hydrophilic. Therefore, the state of the conductive layer, for example, the organic contained in the conductive layer Regardless of the type of binder component, the surface of the conductive layer can be reliably hydrophilized. Therefore, the range of selection of the type of organic binder component can be expanded.

  The surfactant is not particularly limited, but is an anionic surfactant such as fatty acid salt, α-sulfo fatty acid ester salt, alkylbenzene sulfonate, alkyl sulfate, alkyl ether sulfate, alkyltrimethylammonium salt, dialkyldimethyl. Examples include cationic surfactants such as ammonium chloride and alkylpyridinium chloride, amphoteric surfactants such as alkylcarboxybetaine, and nonionic surfactants such as fatty acid diethanolamide and polyoxyethylene glycols. 1 type (s) or 2 or more types are used in combination. When two or more types are used in combination, it is preferable to use a combination of an anionic surfactant and a nonionic surfactant, or a combination of a cationic surfactant and a nonionic surfactant.

Moreover, when using combining an ionic surfactant and a nonionic surfactant, it is preferable that the mixing ratio is about 1: 1 to 1: 5 by weight ratio, and is 1: 1 to 1: 4. More preferred is the degree.
Moreover, the solvent which melt | dissolves surfactant is not specifically limited, Various organic solvents can be used.

Moreover, it is preferable that the content rate of surfactant in a process liquid is about 0.1-0.5 wt%, and it is more preferable that it is about 0.2-0.4 wt%. As a result, a surface layer containing a surfactant at a necessary and sufficient density can be formed on the conductive layer. As a result, a highly hydrophilic electrode can be obtained.
In addition, the various coating methods mentioned above are used for the method of apply | coating a process liquid.
The surface layer thus formed on the conductive layer preferably has an average thickness of about 0.01 to 1 μm, and more preferably about 0.05 to 0.5 μm. By setting the thickness of the surface layer within the above range, the hydrophilicity of the surface layer is surely expressed regardless of the surface state of the conductive layer.

By subjecting the conductive layers 201, 202, 203, the conductive layers 601, 602, 603, and the conductive layers 701, 702, 703 to the surface treatment as described above, the respective surfaces become hydrophilic.
The conductive layers 601, 602, and 603 that have been subjected to the surface treatment become the counter electrodes 61, 62, and 63.
In addition, the conductive layers 701, 702, and 703 that have undergone surface treatment become reference electrodes 71, 72, and 73.

[4] Next, as illustrated in FIG. 6D, reaction layers 41, 42, and 43 are formed on the conductive layers 201, 202, and 203 that have been subjected to the surface treatment.
Hereinafter, the case where each reaction layer 41, 42, 43 is configured to include receptors 411, 421, 431 in a matrix composed of polymers 413, 423, 433 will be described as an example.
First, each of the receptors 411, 421, 431 and the polymers 413, 423, 433 are dissolved in a solvent to prepare a liquid material.
Moreover, you may add a crosslinking agent and the mediators 412, 422, and 432 in a liquid material as needed.
The content of each component in the liquid material is set to a ratio such that the content of each component in the reaction layers 41, 42, and 43 falls within the above range.

Next, this liquid material is supplied to the sample supply space 5.
At this time, in step [3], the surfaces of the conductive layers 201, 202, and 203 are hydrophilized, so that the liquid material is uniformly spread on the surfaces.
Thereafter, the reaction layers 41, 42, and 43 are obtained by leaving them to stand for a certain period of time, followed by washing and drying.
Here, the temperature of the liquid material and the temperature of the atmosphere are each preferably about 3 to 40 ° C., more preferably about 10 to 25 ° C.
Further, the standing time is preferably about 10 to 180 minutes, and more preferably about 20 to 60 minutes.

The reaction layers 41, 42, 43 obtained in this way obtain a uniform and homogeneous surface layer when the liquid material is uniformly spread on the surfaces of the conductive layers 201, 202, 203 when supplying the liquid material. be able to. The surface layer has high adhesion to the surfaces of the conductive layers 201, 202, and 203, respectively.
Then, the working electrodes 21, 22, and 23 are obtained by securely fixing the reaction layers 41, 42, and 43 on the conductive layers 201, 202, and 203.
The sensor 1 is obtained by the above process.

  In the sensor 1 thus obtained, the reaction layers 41, 42, and 43 are formed on the conductive layers 201, 202, and 203 with good adhesion. Electrons generated by the reaction between 52 and 53 and the receptors 411, 421, and 431 can be reliably moved to the conductive layers 201, 202, and 203 side. That is, the working electrodes 21, 22, and 23 having excellent electron mobility are obtained.

  Further, since the counter electrodes 61, 62, 63 and the reference electrodes 71, 72, 73 are each composed of a conductive layer having a hydrophilic surface, the liquid sample 50 has good wettability with respect to these electrodes ( Adhesion). Therefore, it is possible to obtain the sensor 1 which can reliably perform the functions of these electrodes and can measure the current values taken out by the working electrodes 21, 22, and 23 with high accuracy and high sensitivity.

Further, when the sensor 1 includes a plurality of detection units 11, 12, and 13, there is an advantage that variation in the detected current value between the detection units is reduced due to the above-described improvement in adhesion. Thereby, it is possible to obtain a multisensor with a small measurement error of current values obtained between a plurality of detection units.
Further, by providing such a sensor 1, a detection apparatus capable of detecting a detection target with high accuracy and high sensitivity can be obtained.

As mentioned above, although the manufacturing method and sensor of the sensor of this invention were demonstrated based on embodiment of illustration, this invention is not limited to these.
For example, in the method for manufacturing a sensor of the present invention, one or more optional steps may be added as necessary.
Further, for example, one or more layers for any purpose (improvement of adhesion) may be provided between the layers of the sensor 1 within a range that does not cause deterioration of the sensor characteristics.

Next, specific examples of the present invention will be described.
1. Production of sensor (Example 1)
<1A> First, Au powder (average particle diameter: 2 μm), polyacrylate resin, and ethanol (dispersion medium) were mixed to prepare a dispersion.
<2A> Next, a glass substrate was prepared, and a dispersion was supplied onto the substrate by a droplet discharge method to form 16 sets of liquid films having the shape of a working electrode and a counter electrode. In addition, a liquid film having the shape of a wiring for electrically connecting these electrodes and an external processing circuit was also formed.

<3A> Next, Ag-AgCl powder (average particle diameter 2 μm), polyacrylate resin, and ethanol (dispersion medium) were mixed to prepare a dispersion.
<4A> Next, on the glass substrate on which the liquid film is formed in the step <2>, the liquid dispersion obtained in the step <3> is supplied to form 16 liquid films that form the shape of the reference electrode. Formed.
<5A> Next, by leaving these liquid coatings to stand, the ethanol is volatilized and removed, and a conductive layer corresponding to 16 working electrodes, a conductive layer corresponding to 16 counter electrodes, and 16 references. Conductive layers and wirings corresponding to the electrodes were formed. The average thickness of these electrodes and wirings was 200 μm.

<6A> Next, a resin substrate having a circular opening opened in the vicinity of each electrode was prepared, and this resin substrate was bonded onto a glass substrate with an adhesive. Thereby, a partition was formed around each conductive layer, and a sample supply space was formed.
<7A> Next, a mask was provided on the glass substrate so as to cover a region other than the formation region of each conductive layer. Each conductive layer was irradiated with ultraviolet light having a wavelength of 254 nm for 3 minutes in an oxygen-existing atmosphere. Thereby, a counter electrode and a reference electrode were obtained.

<8A> Next, glucose oxidase (receptor) was dissolved in pure water to prepare an enzyme-containing solution. The concentration of the enzyme-containing solution was 1 mM.
<9A> Next, the enzyme-containing solution was supplied onto the conductive layer corresponding to the working electrode and allowed to stand at 23 ° C. for 1 hour. Thereafter, it was washed with pure water and dried by nitrogen blowing. Thereby, a working electrode was obtained and a sensor was produced.
<10A> Next, each wiring of the sensor was connected to a computer equipped with a processing circuit.

(Example 2)
<1B> First, in the same manner as in Steps <1A> to <4A> of Example 1, a conductive layer corresponding to 16 working electrodes, a conductive layer corresponding to 16 counter electrodes on a glass substrate, 16 Conductive layers and wirings corresponding to the reference electrodes were formed.
<2B> Next, the reaction amplifying agent represented by the chemical formula (2) was dissolved in dimethyl sulfoxide containing triethylamine to prepare a treatment solution. Note that the concentration of the reaction amplifying agent in the treatment liquid was 0.5 wt%, and the temperature of the treatment liquid was 23 ° C. In the chemical formula (2), R ³ represents (CF ₂ ) ₅ CF ₃ and R ⁴ represents CH ₃ . And this processing liquid was supplied on each conductive layer by the inkjet method.

<3B> Next, in this state, it was left at a temperature of 23 ° C. for 2 hours. Thereafter, the substrate was washed with pure water and dried by nitrogen blowing. Thereby, a self-assembled film containing a reaction amplification agent on each conductive layer was obtained.
<4B> Next, a mask was provided on the glass substrate so as to cover a region other than the region where each conductive layer was formed. Each conductive layer was irradiated with ultraviolet light having a wavelength of 365 nm for 1 minute. Thereby, a counter electrode and a reference electrode were obtained.
<5B> Next, in the same manner as steps <8A> to <9A> in Example 1, a reaction layer was formed on the conductive layer corresponding to the working electrode, thereby obtaining the working electrode and producing a sensor. . And the sensor was connected to the computer like process <10A>.

(Example 3)
<1C> First, in the same manner as in steps <1A> to <4A> of Example 1, a conductive layer corresponding to 16 working electrodes, a conductive layer corresponding to 16 counter electrodes on a glass substrate, 16 Conductive layers and wirings corresponding to the reference electrodes were formed.
<2C> Next, alkylbenzene sulfonate (anionic surfactant) and polyoxyethylene glycol (nonionic surfactant) were dissolved in pure water to prepare a treatment solution. The mixing ratio of alkylbenzene sulfonate and polyoxyethylene glycol in this treatment liquid was 1: 3 by weight. And this processing liquid was supplied on each conductive layer by the inkjet method.

<3C> Next, in this state, it was left at a temperature of 23 ° C. for 2 hours. Thereafter, the substrate was washed with pure water and dried by nitrogen blowing. Thereby, a surface layer containing a surfactant was obtained on each conductive layer.
<4C> Next, in the same manner as steps <8A> to <9A> in Example 1, a reaction layer was formed on the conductive layer corresponding to the working electrode, thereby obtaining the working electrode and producing a sensor. . And the sensor was connected to the computer like process <10A>.

Example 4
<1D> First, in the same manner as in steps <1A> to <4A> of Example 1, a conductive layer corresponding to 16 working electrodes, a conductive layer corresponding to 16 counter electrodes on a glass substrate, 16 Conductive layers and wirings corresponding to the reference electrodes were formed.
<2D> Next, denatured bovine serum albumin (denatured BSA) was dissolved in a phosphate buffer solution (PBS buffer solution) having a pH of 7.3 to prepare a treatment solution. The content of denatured bovine serum albumin in this treatment liquid was 3 wt%, and the temperature of the liquid material was 23 ° C. And this processing liquid was supplied on each conductive layer by the inkjet method.

<3D> Next, in this state, it was left at a temperature of 23 ° C. for 2 hours. Thereafter, the substrate was washed with pure water and dried by nitrogen blowing. Thereby, a surface layer containing denatured bovine serum albumin was obtained on each conductive layer.
<4D> Next, a reaction layer was formed on the conductive layer corresponding to the working electrode in the same manner as steps <8A> to <9A> in Example 1, thereby obtaining a working electrode and producing a sensor. . And the sensor was connected to the computer like process <10A>.

(Example 5)
<1E> First, in the same manner as in Steps <1A> to <4A> of Example 1, a conductive layer corresponding to 16 working electrodes, a conductive layer corresponding to 16 counter electrodes on a glass substrate, 16 Conductive layers and wirings corresponding to the reference electrodes were formed.
<2E> Next, lipase (lipolytic enzyme) was dissolved in a mixed solvent of a phosphate buffer solution (PBS buffer solution) having a pH of 7.3 and dimethylformamide to prepare a treatment solution. In addition, the content rate of lipase in this process liquid was 3 wt%, and the content rate of dimethylformamide was 15 vol%. The temperature of the treatment liquid was 37 ° C. And this processing liquid was supplied on each conductive layer by the inkjet method.
<3E> Next, in this state, it was left at a temperature of 37 ° C. for 2 hours. Thereafter, the substrate was washed with pure water and dried by nitrogen blowing. Thereby, a counter electrode and a reference electrode were obtained.
<4E> Next, a reaction layer was formed on the conductive layer corresponding to the working electrode in the same manner as in Steps <8A> to <9A> of Example 1, thereby obtaining a working electrode and producing a sensor. . And the sensor was connected to the computer like process <10A>.
(Comparative example)
A sensor was produced in the same manner as in Example 1 except that the step <7A> in Example 1 was omitted, and each wiring of the sensor was connected to a computer.

2. Evaluation First, a liquid sample containing a glucose substrate was prepared. The concentration of the substrate in the liquid sample was 5 mM.
Next, while supplying this liquid sample to 16 detection parts of the sensor obtained in each example and comparative example, between the working electrode and the counter electrode, and between the reference electrode and the counter electrode A voltage of 0.5 V was applied.
Next, the current value taken out from each working electrode was measured.

As a result, the current value measured by the sensor obtained in each example was larger than the current value measured by the sensor obtained in the comparative example.
Further, when variations in current values taken out from a plurality of working electrodes were compared, in the sensors obtained in the respective examples, the variations in current values were smaller than in the sensors obtained in the comparative examples.

It is a schematic diagram (perspective view) which shows the structure of the detection apparatus of this invention provided with the sensor of this invention. It is a top view which shows typically the sensor of this invention. It is a top view which expands and shows one detection part with which the sensor shown in FIG. 1 is provided. FIG. 3 is a cross-sectional view of the sensor shown in FIG. It is the elements on larger scale of sectional drawing shown in FIG. It is a longitudinal cross-sectional view for demonstrating the manufacturing method of the sensor shown in FIG. It is a longitudinal cross-sectional view for demonstrating the manufacturing method of the sensor shown in FIG.

Explanation of symbols

  DESCRIPTION OF SYMBOLS 1 ... Sensor 2 ... Board | substrate 3 ... Wiring 5 ... Sample supply space 6 ... Bulkhead 11, 12, 13 ... Detection part 201,202,203,601,602,603,701,702,703 ... Conductive layer 21, 22, 23 ... Working electrode 41, 42, 43 ... Reaction layer 411, 421, 431 ... Receptor 412, 422, 432 ... Mediator 413, 423, 433 ... Polymer 50 ... Liquid Sample 51, 52, 53 ... Target 61, 62, 63 ... Counter electrode 71, 72, 73 ... Reference electrode 100 ... Measuring device (detection device) 101 ... Processing circuit 102 ... Arithmetic device 103 ... Connector 104 Wiring

Claims (20)

A first step of forming a conductive layer containing conductive material particles and an organic binder component on a substrate;
A second step of hydrophilizing the surface of the conductive layer by performing at least one of a process of decomposing the organic binder component in the conductive layer and a process of introducing a hydrophilic group onto the conductive layer. A method for producing a sensor, comprising:   Furthermore, the manufacturing method of the sensor of Claim 1 which has a 3rd process of forming the reaction layer which reacts with the detection target object in a sample on the said conductive layer which performed the said 2nd process.   The sensor manufacturing method according to claim 2, wherein the reaction layer includes an oxidoreductase that performs an electrochemical reaction with the detection target.   In the first step, a liquid material containing the conductive material particles and the organic binder component in a solvent is supplied onto the substrate to form a liquid film, and the liquid film is formed from the liquid film. The method for manufacturing a sensor according to claim 1, wherein the conductive layer is formed by removing the solvent.   The sensor manufacturing method according to claim 4, wherein the liquid material is supplied by a droplet discharge method.   The method for manufacturing a sensor according to claim 1, wherein the treatment for decomposing the organic binder component is to irradiate the conductive layer with ultraviolet light.   The method of manufacturing a sensor according to claim 6, wherein the wavelength of the ultraviolet light is 200 to 300 nm.   The sensor manufacturing method according to claim 6, wherein the organic binder component includes a polyacrylate resin or an epoxy resin.   The sensor manufacturing method according to any one of claims 1 to 5, wherein the treatment for decomposing the organic binder component comprises bringing the conductive layer into contact with an enzyme that decomposes fat.   The method for producing a sensor according to claim 9, wherein the enzyme that degrades fat is lipase.   The sensor manufacturing method according to claim 9 or 10, wherein the organic binder component includes a resin containing an ester bond.   On the conductive layer, by forming a surface layer containing a reaction amplifying agent having a property that the decomposability by irradiation with ultraviolet light is greater than that of the organic binder component, and then irradiating the surface layer with ultraviolet light, The method for producing a sensor according to claim 1, wherein a hydrophilic group is introduced on the surface layer.   The method for producing a sensor according to claim 12, wherein the reaction amplification agent contains a nitrobenzyl derivative.   The method of manufacturing a sensor according to claim 12 or 13, wherein the wavelength of the ultraviolet light is 300 to 400 nm.   In the second step, a hydrophilic group is introduced onto the conductive layer by forming a surface layer containing a polypeptide chain or a modified product thereof on the conductive layer. A manufacturing method of the sensor.   The method for producing a sensor according to claim 15, wherein the polypeptide chain contains bovine serum albumin or a modified product thereof.   The sensor according to any one of claims 1 to 5, wherein in the second step, a hydrophilic group is introduced onto the conductive layer by forming a surface layer containing a surfactant on the conductive layer. Production method.   A sensor manufactured by the method for manufacturing a sensor according to claim 1. A substrate;
A counter electrode provided with a conductive layer provided on the substrate and containing particles of a conductive material and an organic binder component;
A working electrode comprising a conductive layer containing conductive material particles and an organic binder component provided on the substrate, and a reaction layer provided on the conductive layer and reacting with an object to be detected in a sample. And
At least one of the conductive layers is subjected to at least one of a treatment for decomposing the organic binder component to be contained and a treatment for introducing a hydrophilic group on the surface.   A detection apparatus comprising the sensor according to claim 18.

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