WO2017082253A1 - Sensor - Google Patents

Abstract

The present invention addresses the problem of providing a compact sensor with which it is possible to perform simultaneous measurements of glucose and biologically relevant substances other than glucose. The present invention pertains to a sensor characterized by having at least: a first detection element for detecting glucose in a biological liquid; and a second detection element for detecting biologically relevant substances other than glucose, the second detection element including a semiconductor element.

Description

Sensor

The present invention relates to a sensor capable of simultaneously measuring a plurality of biological substances.

In order to conduct screening tests and treatments for diabetes, glycated hemoglobin (hereinafter also referred to as HbA1c), glycated albumin (hereinafter referred to as glycoalbumin) and the like, which are important together with glucose in blood, are measured.

In diabetes, the production of glycated protein is enhanced, and the concentration of HbA1c contained in erythrocytes and glycoalbumin in serum reflect the average blood glucose level over a certain period in the past. Glucose shows the current blood glucose state, whereas glycoalbumin reflects the blood glucose state about 1 to 2 weeks ago, and HbA1c reflects the blood glucose state about 1 to 2 months ago. Therefore, measurement of these glycated proteins is important for the time-dependent diagnosis or symptom management of diabetes symptoms in that the blood glucose concentration can be determined over time. Therefore, as an index for diabetes management, there is a demand for a rapid and accurate quantitative method for glucose concentration and HbA1c or glycoalbumin.

As a method for measuring glucose in a biological fluid, for example, there is an electrochemical detection method called an enzyme electrode method. In this method, information correlated with the glucose concentration in the biological sample is output to an electrode in contact with the biological sample, and the glucose concentration is calculated based on this output. Since the enzyme electrode method can be applied to a small blood glucose level sensor, it is mainly used by diabetic patients all over the world.

On the other hand, as a method for measuring HbA1c, for example, a chromatography method such as HPLC, an immunoassay method using an antibody such as latex immunoagglutination, and an enzyme method using an enzyme that acts on a glycated protein are known. Both are widely used in clinical settings as detection methods using light.

In clinical practice, some devices for measuring both glucose and glycohemoglobin (glycated hemoglobin containing HbA1c) have been introduced. Examples of such a device include a device in which devices for measuring glucose and glycated hemoglobin are connected, and a device in which both devices can be measured (see, for example, Patent Document 1).

Japanese Laid-Open Patent Publication No. 2014-95715

In the technique described in Patent Document 1, an increase in the size of the apparatus is suppressed by sharing various elements for measuring glucose and glycohemoglobin. However, in each measurement mechanism, for example, it is difficult to further reduce the size of the apparatus because a light source is necessary.

In view of the above problems, an object of the present invention is to provide a small sensor capable of simultaneously measuring glucose and biologically related substances other than glucose such as HbA1c and glycoalbumin.

In order to solve the above problems, the present invention has the following configuration.
1. It has at least a first detection element that detects glucose in a biological fluid and a second detection element that detects a biological substance other than glucose, and the second detection element includes a semiconductor element. Sensor.
2. 2. The sensor according to 1 above, wherein the biological substance is glycated hemoglobin or glycated albumin.
3. 3. The sensor according to 1 or 2, further comprising a third detection element, wherein the third detection element detects hemoglobin.
4). 4. The sensor according to 3 above, wherein the third detection element includes a semiconductor element.
5). 5. The sensor according to any one of 1 to 4, wherein a detection part area of the second detection element is twice or more a detection part area of the third detection element.
6). 6. The sensor according to any one of 1 to 5, wherein the semiconductor element contains a carbon nanotube.
7). The semiconductor element includes at least a substrate, a first electrode, a second electrode, and a semiconductor layer, and the first electrode and the second electrode each include a metal type of 80 wt% or more, a double-walled carbon nanotube, 7. The sensor according to any one of 1 to 6, containing at least one selected from multi-walled carbon nanotubes.
8). 8. The sensor according to any one of 1 to 7, wherein the semiconductor layer contains carbon nanotubes.
9. 9. The sensor according to any one of 6 to 8, wherein a conjugated polymer is attached to at least a part of the surface of the carbon nanotube.
10. 10. The sensor according to any one of 7 to 9, wherein another biological substance that selectively interacts with the biological substance is immobilized on the semiconductor layer.
11. 11. The sensor according to any one of 1 to 10, wherein the sensor has a biological fluid inlet, and a path connecting the inlet to the first detection element and the second detection element.
12 12. The sensor according to any one of 1 to 11, wherein the biological fluid is blood.
13. The sensor according to any one of 1 to 12, wherein the path has a space for mixing the blood and a hemolytic agent between the blood inlet and the second detection element.

According to the present invention, a small sensor capable of simultaneously measuring a plurality of biological substances can be provided.

FIG. 1 is a perspective view showing an embodiment of the sensor of the present invention. FIG. 2 is a schematic cross-sectional view showing an example of a semiconductor element. FIG. 3 is a schematic cross-sectional view showing an example of a semiconductor element. FIG. 4 is a schematic cross-sectional view showing an example of a semiconductor element. FIG. 5A is a schematic cross-sectional view illustrating an example of a semiconductor element. FIG. 5B is a cross-sectional view taken along line CC ′ of FIG. 5A. FIG. 6A is a schematic cross-sectional view illustrating an example of a semiconductor element. 6B is a cross-sectional view taken along line DD ′ of FIG. 6A. FIG. 7 is a schematic cross-sectional view showing an example of a semiconductor element. FIG. 8 is a perspective view showing an embodiment of the sensor of the present invention. FIG. 9 is a perspective view showing an embodiment of the sensor of the present invention.

The sensor of the present invention has at least a first detection element for detecting glucose in a biological fluid and a second detection element for detecting a biological substance other than glucose, and the second detection element is a semiconductor. Including elements.

The sensor of the present invention will be described with reference to FIG. 1 showing an embodiment thereof. FIG. 1 is a schematic perspective view of a sensor having a first detection element 401 and a second detection element 402. The two detection elements are both formed on the substrate 10 and are bonded to the upper substrate 20 to form a chip.

(First detection element)
The first detection element 401 detects glucose in the biological fluid. The first detection element 401 includes at least a pair of electrodes 101 on the substrate 10 and a reaction layer 110 between the pair of electrodes.
The pair of electrodes are detection electrodes and are electrically connected to the connection portion 102 and the wiring 107. Using this connection, a voltage can be applied between the electrodes from a power source connected to the connection part, or an electric signal generated in the reaction layer can be taken out from the connection part.

The reaction layer contains an enzyme that reacts with glucose. When glucose in the biological fluid reacts with the enzyme in the reaction layer, the glucose is decomposed and further oxidized to produce hydrogen peroxide. Since the current when hydrogen peroxide is decomposed depends on the blood glucose concentration, the amount of current is detected as the glucose concentration.

The enzyme that reacts with glucose is not particularly limited, and examples thereof include glucose oxidase (GOD) and glucose dehydrogenase (GDH).

In addition, the reaction layer may contain a mediator in addition to the above enzyme. The mediator temporarily receives electrons generated when glucose is decomposed, and then emits electrons when a voltage is applied between the pair of electrodes. Since the amount of current generated thereby depends on the blood glucose concentration, the amount of current is detected as the glucose concentration.

Examples of the mediator include various ions, and specific examples include ions generated from ferricyanide salts, ferrocene and derivatives thereof, methylene blue, benzoquinone and derivatives thereof, naphthoquinone, phenazine methosulfate, and thionine. Particularly preferred is ferricyan ion derived from potassium ferricyanide. When the ferricyan ion receives the electron, it is converted into ferrocyanide ion, and then returns to ferricyan ion again by applying a voltage.

The method using this detection element is generally called an electrode method using electrochemistry and is the mainstream of blood glucose level measuring instruments in recent years. Depending on the enzyme to be reacted, there are a glucose oxidase method (GOD method), a glucose dehydrogenase method (GDH method) and the like, and any enzyme may be used as long as it is an electrode method. Advantages of the electrode method include a short time until measurement, a small amount of blood to be collected, and a simple and easy-to-understand apparatus.

Examples of the material used for the substrate include inorganic materials such as silicon wafer, glass, and alumina sintered body, polyimide, polyester, polycarbonate, polysulfone, polyethersulfone, polyethylene, polyphenylene sulfide, polyparaxylene, aliphatic polyester, Examples include, but are not limited to, organic materials such as polyethylene terephthalate, polybutylene terephthalate, polypropylene, polyvinyl alcohol, polyvinyl chloride, polyvinylidene fluoride, polysiloxane, polyvinylphenol, and polyaramide, or a mixture of inorganic material powder and organic material. Is not to be done. These materials may be used alone, or a plurality of materials may be laminated or mixed.

Examples of the material used for the electrode include conductive metal oxides such as tin oxide, indium oxide, and indium tin oxide (ITO), or platinum, gold, silver, copper, iron, tin, zinc, aluminum, indium, Metals such as chromium, lithium, sodium, potassium, cesium, calcium, magnesium, palladium, molybdenum, amorphous silicon, and polysilicon and their alloys, inorganic conductive materials such as copper iodide and copper sulfide, polythiophene, polypyrrole, Polyaniline, organic conductive materials such as polyethylenedioxythiophene and polystyrenesulfonic acid complexes, glassy carbon, amorphous carbon, graphite, carbon fiber, and other carbon materials such as diamond, carbon nanotube, and graphene Nano-carbon material, such as, and the like conductive carbon black, but is not limited thereto.

These electrode materials may be used alone, or a plurality of materials may be laminated or mixed. When used as a sensor, the electrode is preferably selected from gold, platinum, palladium, an organic conductive material, and a nanocarbon material from the viewpoint of stability to an aqueous solution in contact with the sensor.

The electrode may be in direct contact with the substrate, or may have an adhesive layer between the electrode and the substrate. In the case where the adhesive layer also serves as an insulating layer that can be electrically insulated, a metallic or semiconducting substance can be used as the substrate.

The material used for the connecting portion may be any conductive material that can generally be used as an electrode. Specific examples include, but are not limited to, carbon materials, metals, alloys, various compounds of metals and alloys (eg, oxides, hydroxides, halides, sulfides, nitrides, and carbides). It is not something.

Preferably, a carbon material, platinum, palladium, gold, silver, aluminum or the like is used. Examples of the carbon material include CNT, graphite, pyrolytic carbon, glassy carbon, acetylene black, and carbon black.

The material used for the wiring may be any conductive material that can be generally used as an electrode. Specific examples include, but are not limited to, carbon materials, metals, alloys, various compounds of metals and alloys (eg, oxides, hydroxides, halides, sulfides, nitrides, and carbides). It is not something.

Preferably, a carbon material, platinum, palladium, gold, silver, aluminum or the like is used. Examples of the carbon material include CNT, graphite, pyrolytic carbon, glassy carbon, acetylene black, and carbon black.

(Second detection element)
The second detection element 402 detects a biological substance other than glucose. The second detection element 402 includes a semiconductor element including at least a first electrode 103 and a second electrode 104 on the substrate 10 and a semiconductor layer 111 between the electrodes. The first electrode and the second electrode are detection electrodes, and are electrically connected to the connection portion 102 and the wiring 107. Although not shown, a power source and a detection unit are connected to the connection unit 102. Using this connection, a voltage can be applied between the electrodes from the power source, and an electrical signal generated in the semiconductor layer can be taken out from the connection portion.

When a biological substance other than glucose is disposed in the vicinity of the semiconductor layer, the current value or the electric resistance value flowing between the first electrode and the second electrode changes. By measuring the change, the biological substance can be detected. By changing the substance that selectively interacts with the biological substance contained in the semiconductor layer, various biological substances can be detected.

Advantages of a detection element using a semiconductor element include a short time until measurement, a small amount of blood to be collected, and a simple and easy-to-understand apparatus. In particular, the FET type detection element is preferable in that labeling with a phosphor or the like is unnecessary, electrical signal conversion is fast, and connection with an integrated circuit is easy.

The first detection element has the same characteristics and size as the sensor chip of the blood glucose sensor, and is therefore equivalent to the current chip size and small. Further, the second detection element has the same size, but has a channel and is slightly larger.

The size of the sensor having these two is at least larger than the current blood glucose level sensor chip size, but is not as large as the device described in Patent Document 1. As described above, the measurement of glucose is an electrochemical method mainly using an electrode method, and the measurement of biological materials other than glucose is an electrical method using a semiconductor element. Both of them are preferable in that they do not use light, so that electrical signal conversion is fast and integration is easy.

In FIG. 1, the first detection element and the second detection element are provided with a common substrate 10, but may be provided with different substrates. The same applies to the upper substrate 20.
In addition, as the material used for the upper substrate 20, the same material as that used for the substrate 10 can be used. In one sensor, the material of the substrate 10 and the upper substrate 20 may be the same or different.

(Third detection element)
In the present invention, as shown in FIGS. 8 and 9, a third detection element 403 may be provided in addition to the first and second detection elements. The third detection element 403 detects hemoglobin. As shown in FIG. 8, the third detection element 403 includes at least (I) a pair of electrodes 121 on the substrate 10 and a reaction layer 112 between the pair of electrodes, or as shown in FIG. (II) An element having the first electrode 122 and the second electrode 123 and the semiconductor element 202 including the semiconductor layer 201 between these electrodes.

Any method may be used in the present invention. Here, the ratio of HbA1c or the ratio of glycohemoglobin to the total hemoglobin can be calculated from the total hemoglobin concentration in the sample.

Concentrations of total hemoglobin include known methods such as the methemoglobin method, cyan methemoglobin method, azide methemoglobin method, sodium dodecyl sulfonate method, alkali hematin method, green chromophore formation method, and oxyhemoglobin method. . Among them, for the measurement of hemoglobin, an electrochemical method in which a reagent such as potassium ferricyanide is reacted and a method using light by an antigen-antibody reaction using an anti-hemoglobin antibody as an antibody have been established.

However, a specific method of hemoglobin detection using the semiconductor element described in the second detection element has not been reported so far, and it has been detected for the first time by the present invention. In the second detection element and the third detection element, from the viewpoint of hemolysis of blood, which will be described later, and from the viewpoint of manufacturing the sensor chip, as shown in FIG. 9, (II) the first electrode 122 and the second detection element An element 202 having a semiconductor element including the electrode 123 and the semiconductor layer 201 between these electrodes is preferable.

Further, it is preferable that the detection area of the second detection element is twice or more than the detection area of the third detection element. This is because the hemoglobin detected by the third detection element is present in a large amount in the blood, and the glycated hemoglobin or glycated albumin, which is a glycated product thereof, is at most 50% or less. Therefore, if the detection area of the second detection element is twice or more larger than the third detection area, the detection reliability of hemoglobin and its saccharified product is improved. More preferably, they are 2 times or more and 10 times or less, More preferably, they are 2 times or more and 5 times or less, More preferably, they are 2 times or more and 3 times or less, Especially preferably, they are 2 times or more and 2.5 times or less.

(Inlet and path)
The sensor of the present invention preferably has a biological fluid inlet 301 and a path 302 that connects the inlet to the first detection element 401, the second detection element 402, and the third detection element 403, respectively. . With this route, it is possible to deliver the biological fluid to each of the first detection element, the second detection element, and the third detection element by one injection of the biological fluid, and the operability is excellent. Further, since it is only necessary to collect the biological fluid once, the amount of biological fluid collected can be reduced. Furthermore, it is possible to perform measurement using biological fluid in the same state, and it is possible to reduce as much as possible time variation of biological fluid and errors between measurements due to collection methods.

The injection port is provided in any part of the sensor body. Although it is provided on the upper surface of the upper substrate 20 in FIG. 1, the present invention is not limited thereto, and may be the side surface of the upper substrate 20 or the side surface of the substrate 10.

As shown in FIG. 1, in the substrate 10, a depression 304 may be provided at a location corresponding to the injection port 301 provided in the upper substrate 20.

The path is connected from the inlet to the first detection element and the second detection element. In FIG. 1, the path is provided in a groove shape on the substrate 10, and the biological fluid injected from the injection port 301 accumulates in the depression 304 and flows from there to the path 302.

At this time, a groove having substantially the same shape as the path 302 may be provided at a position corresponding to the path 302 on the bonding surface of the upper substrate 20 to the substrate 10. Thus, the size of the groove can be increased when the upper substrate 20 and the substrate 10 are bonded together.

Also, the positional relationship between the inlet and the route is not limited to these. For example, a path may be provided in a tubular shape inside the upper substrate 20 so as to be digged from the inlet toward the first detection element and the second detection element. A tubular route and a groove-like route may be used in combination. If the position of the injection port changes, the position where the path is provided also changes.

The injection port and path are made by processing the substrate. The processing method greatly depends on the material. For example, if silicon or glass is used, fine processing using photolithography, laser processing, and the like are possible. In addition to these, if it is plastic, injection molding, imprinting, hot embossing, drilling and the like are also possible. However, in any case, the processing method is not particularly limited to these.

The shape of the channel is typically a groove shape or a tubular shape, but is not limited to these as long as it can flow a biological fluid.
After processing the substrate, the processed surface may be processed so that the biological fluid can be easily flowed.
The width of the path is not particularly limited, but is preferably about 1 μm to 1 mm. The depth of the path is not particularly limited, but is preferably about 1 μm to 1 mm.

Examples of a method for bonding the substrate 10 and the upper substrate 20 include a method using an adhesive. Any adhesive may be used as long as it can form a liquid layer. For example, an ultraviolet curing type, a thermosetting type, and a two-component mixed type adhesive may be used. In consideration of the affinity with the substrate, an adhesive that can be applied uniformly with a thickness of about several micrometers is preferable. For example, if the substrate is a hydrophilic glass substrate, the adhesive is also preferably hydrophilic.

When the substrate or the upper substrate is a glass substrate, as another method of bonding them together, a method of welding the inner surfaces of them using a laser and integrating them is also mentioned.
Various methods, such as capillary action, pumping, pressurization, and centrifugation, can be used to cause the biological fluid to flow through the path.

(Biological fluid)
A biological fluid is a fluid that an organism has in the body in some form. Specific examples include blood, lymph, tissue fluid, body cavity fluid, digestive fluid, sweat, tears, runny nose, urine, semen, vaginal fluid, amniotic fluid, spinal fluid, synovial fluid, cell suspension and milk as they are. Can be used. Moreover, the sample which crushed or removed the cell component etc. previously from the biological sample may be sufficient. Examples of the digestive juice include saliva, gastric juice, bile, pancreatic juice, and intestinal juice.

As a sample to be used for the sensor including the semiconductor element of the present invention, among biological fluids, blood, saliva, sweat, tears, urine and the like are preferable because of easy availability, and among them blood is more preferable because it contains a lot of biological information.

(Biological substances)
The biological substance other than glucose is not particularly limited as long as it is detected by the second detection element, and any substance can be used. Specifically, enzyme, antigen, antibody, hapten, hapten antibody, peptide, oligopeptide, polypeptide (protein), hormone, nucleic acid, oligonucleotide, biotin, biotinylated protein, avidin, streptavidin, lipid, steroid, sugar Saccharides such as oligosaccharides and polysaccharides (excluding glucose), low molecular compounds, high molecular compounds, inorganic substances and complexes thereof, viruses, bacteria, cells, living tissues, and substances constituting these.

These are either hydroxyl groups, carboxy groups, amino groups, mercapto groups, sulfo groups, phosphonic acid groups, organic or inorganic salts thereof, functional groups such as formyl groups, maleimide groups and succinimide groups, or biologically related substances. This reaction or interaction causes a change in the electrical characteristics of the semiconductor layer in the sensor of the present invention.

The low molecular weight compound is not particularly limited, and examples thereof include a gaseous compound at normal temperature and normal pressure such as ammonia and methane emitted from a living body and a solid compound such as uric acid. Preferably, solid compounds such as uric acid are used.

Among these, proteins, viruses, and bacteria are preferable as the sensing target substance. Examples of proteins include PSA, hCG, IgE, BNP, NT-proBNP, AFP, CK-MB, PIVKA II, CA15-3, CYFRA, anti-p53, troponin T, procalcitonin, hemoglobin, HbA1c, glycoalbumin, Examples include apolipoprotein and C-reactive protein (CRP).

Examples of viruses include HIV, influenza virus, hepatitis B virus, and hepatitis C virus. Examples of bacteria include Chlamydia, Staphylococcus aureus, and enterohemorrhagic Escherichia coli.

Among the above, hemoglobin, HbA1c, and glycoalbumin, which are types of polypeptides, are preferable. This is because it can be a disease marker for diabetes, and it is significant to combine with a first detection element capable of detecting glucose.

(Hemolysis)
When the biological fluid used for the measurement according to the present invention is blood, it is necessary to perform hemolysis of the blood. In 1 μL of blood, there are millions of red blood cells and several thousand to 10,000 white blood cells, so that hemoglobin in red blood cells cannot be detected as it is. Therefore, it is necessary to treat the cell membrane of red blood cells by damaging or lysing them by various factors such as physical, chemical, biological, etc. and causing hemoglobin or the like to leak out of the cells. This is called hemolysis.

Physiological factors include various mechanical stresses such as pressure and centrifugal force. Typical hemolysis methods include the method of excessively negative pressure in the syringe during blood collection, the method of exposing to excessive centrifugal force during the centrifugation, and the method of agitating or foaming erythrocyte fluid roughly. is there. In addition, a method of mixing red blood cells with a solution having a low osmotic pressure (hypotonic solution) can also be used. Due to the difference in osmotic pressure, extracellular water continues to flow into the cell through the cell membrane, which is a semipermeable membrane, and finally erythrocytes are ruptured. In this case, water, purified water, buffer solution, and the like can also be hemolytic agents.

Chemical factors include lipid dissolution or damage that constitutes the cell membrane due to various solvents and surfactants. Examples of such solvents and surfactants include alcohols such as methanol and ethanol, various organic solvents other than acetone, and soap.

As a biological factor, hemolysis caused by antibodies and complement is known. Complement activation signal transduction begins when antibodies against erythrocytes bind or by another activation mechanism, and each component of complement is activated sequentially (cascade reaction) and finally penetrates the cell membrane. A channel-like protein complex is formed, and the cell membrane is perforated, causing hemolysis.

The hemolysis method applied to the present invention is not particularly limited, but an osmotic pressure method is preferable from the viewpoint of availability and cost. For example, hemolysis can be achieved by a method such as dilution by adding 2 to 100 times the volume of purified water to the blood volume.

The space for performing the hemolysis treatment is not particularly limited. For example, the hemolysis treatment may be performed before the injection port for injecting blood into the sensor, or may be performed between the injection port and the detection element. , Either or not.

In order to perform hemolysis between the injection port and the detection element, it is preferable that the above-described path has a space for mixing blood and a hemolytic agent between the blood inlet and the second or third detection element. .
For example, in FIG. 1, the blood injected from the injection port and the hemolyzing agent are mixed in the space by allowing the hemolytic agent such as the surfactant, purified water, and buffer solution to exist in the space 303 in advance. The mixed liquid flows into the semiconductor layer.

(Semiconductor element)
The semiconductor element included in the second or third detection element will be described in more detail. One aspect of the semiconductor element used in the present invention includes a substrate, a first electrode, a second electrode, and a semiconductor layer, and the semiconductor layer contains carbon nanotubes (hereinafter referred to as CNT). As another aspect, the semiconductor element further includes a third electrode and an insulating layer, and the third electrode is electrically connected to the first electrode, the second electrode, and the semiconductor layer by the insulating layer. Insulated and arranged.

2 and 3 are schematic cross-sectional views showing examples of semiconductor elements.
In the semiconductor element of FIG. 2, a first electrode 103 and a second electrode 104 are formed on a substrate 10, and a semiconductor layer 111 is disposed between the first electrode 103 and the second electrode 104.

In the semiconductor element of FIG. 3, the third electrode 105 and the insulating layer 106 are formed on the substrate 10, and the first electrode 103 and the second electrode 104 are formed. A semiconductor layer 111 containing CNT is disposed on the substrate. In the semiconductor element of FIG. 3, the first electrode 103, the second electrode 104, and the third electrode 105 correspond to the source electrode, the drain electrode, and the gate electrode, respectively, and the insulating layer 106 corresponds to the gate insulating layer, and functions as an FET. .

Examples of the material used for the substrate 10 include inorganic materials such as silicon wafer, glass, and alumina sintered body, polyimide, polyester, polycarbonate, polysulfone, polyethersulfone, polyethylene, polyphenylene sulfide, polyparaxylene, aliphatic polyester, Examples include, but are not limited to, organic materials such as polyethylene terephthalate, polybutylene terephthalate, polypropylene, polyvinyl alcohol, polyvinyl chloride, polyvinylidene fluoride, polysiloxane, polyvinylphenol, and polyaramide, or a mixture of inorganic material powder and organic material. Is not to be done. These materials may be used alone, or a plurality of materials may be laminated or mixed.

In order to suppress non-specific adsorption of proteins in the measurement sample solution, the surface of the substrate 10 may be processed. For example, hydrophilic groups having no charge such as oligoethylene glycol chain and oligo (3,4-dihydroxyphenylalanine) and hydrophilic groups having both positive and negative charges such as phosphorylcholine group are effective.

When a biological substance other than glucose is detected by selective interaction with another biological substance, which will be described later, in order to manufacture the sensor, a solution in which the biological substance is dissolved in the semiconductor layer In some cases, another sensitive substance is immobilized on the sensitive part. In this case, the other biological substance is selectively immobilized on the sensitive part by suppressing the other biological substance from adhering to other than the sensitive part. Thereby, it is suppressed that biological related substances other than glucose are captured by another biological related substance at a place other than the sensitive part, selective detection in the sensitive part becomes dominant, and detection sensitivity is improved.

Examples of materials used for the first electrode 103, the second electrode 104, and the third electrode 105 include conductive metal oxides such as tin oxide, indium oxide, and indium tin oxide (ITO), platinum, gold, silver, Metals such as copper, iron, tin, zinc, aluminum, indium, chromium, titanium, lithium, sodium, potassium, cesium, calcium, magnesium, palladium, molybdenum, amorphous silicon, and polysilicon, and alloys thereof, copper iodide, And inorganic conductive materials such as copper sulfide, organic conductive materials such as polythiophene, polypyrrole, polyaniline, and polyethylenedioxythiophene and polystyrenesulfonic acid complexes, carbon nanotubes, nanocarbon materials such as graphene, and conductive carbon black Etc. That, without being limited thereto.

The nanocarbon material may be any of single-walled carbon nanotubes, double-walled carbon nanotubes, and multi-walled carbon nanotubes containing a metal type of 80% by weight or more. Among them, it is preferable to use double-walled carbon nanotubes in order to obtain high conductive properties. These electrode materials may be used alone, or a plurality of materials may be laminated or mixed.

When used as a sensor, the first electrode 103 and the second electrode 104 are made of gold, platinum, palladium, organic, from the viewpoints of electrical resistance value, ease of film formation, film stability, and stability to aqueous solutions in contact with the sensor. It is preferably selected from conductive materials and nanocarbon materials. In particular, gold or carbon nanotubes are more preferable because the difference in work function with a semiconductor element containing carbon nanotubes is small, so that low power consumption driving is possible.

The width, thickness, interval, and arrangement of the first electrode 103 and the second electrode 104 are arbitrary. The width is preferably 1 μm to 1 mm, the thickness is preferably 1 nm to 1 μm, and the electrode spacing is preferably 1 μm to 10 mm. Further, the width and thickness of the first electrode 103 and the second electrode 104 may not be the same. The shape of the electrode need not be a rectangular parallelepiped, and may be bent or comb-shaped.

The width, thickness, distance from the semiconductor layer, and arrangement of the third electrode 105 are arbitrary. The width is preferably 1 μm to 1 mm, the thickness is preferably 1 nm to 1 μm, and the distance from the semiconductor layer is preferably 1 μm to 10 cm. For example, an electrode having a width of 100 μm and a thickness of 500 nm is disposed at a distance of 2 mm from the semiconductor layer, but is not limited thereto.

In FIG. 3, the third electrode 105 is arranged in parallel with the second electrode 104, but may be arranged vertically or at any other angle. The shape of the third electrode 105 is not limited to a straight line, and may be a curved line or a curved surface. The third electrode 105 is not limited to being disposed immediately above the substrate, but may be disposed on another member disposed on the substrate.

Examples of the material used for the insulating layer 106 include inorganic materials such as silicon oxide and alumina, and organic polymers such as polyimide, polyvinyl alcohol, polyvinyl chloride, polyethylene terephthalate, polyvinylidene fluoride, polysiloxane, and polyvinylphenol (PVP). A material or a mixture of inorganic material powder and organic polymer material can be used.

The film thickness of the insulating layer 106 is preferably 10 nm or more and 5 μm or less. More preferably, they are 50 nm or more and 3 micrometers or less, More preferably, they are 100 nm or more and 1 micrometer or less. The film thickness can be measured by an atomic force microscope or an ellipsometry method.

The semiconductor layer 111 preferably contains CNT. The semiconductor layer 111 may further include an organic semiconductor or an insulating material as long as the electrical characteristics of the CNT are not impaired.
The thickness of the semiconductor layer 111 is preferably 1 nm to 100 nm. By being within this range, it is possible to sufficiently extract changes in electrical characteristics due to interaction with the sensing target substance as electrical signals. More preferably, they are 1 nm or more and 50 nm or less, More preferably, they are 1 nm or more and 20 nm or less.

As a method for forming the semiconductor layer 111, dry methods such as resistance heating vapor deposition, electron beam, sputtering, and CVD can be used. However, a coating method is used from the viewpoint of manufacturing cost and adaptability to a large area. Is preferred.

The coating method includes a step of forming a semiconductor layer by coating a semiconductor component. Specifically, a spin coating method, a blade coating method, a slit die coating method, a screen printing method, a bar coater method, a mold method, a printing transfer method, a dip pulling method, an ink jet method, and the like can be preferably used. The coating method can be selected according to the properties of the coating film to be obtained, such as control and orientation control. In addition, the formed coating film may be annealed in the air, under reduced pressure, or in an inert gas atmosphere (in a nitrogen or argon atmosphere).

A general FET theory will be described. The current flowing between the source electrode and the drain electrode can be controlled by changing the applied gate voltage. The mobility of the FET can be calculated using the following equation (a).
μ = (δId / δVg) L · D / (W · ε r · ε · Vsd) (a)
However, Id is the current between the source and the drain, Vsd is the voltage between the source and the drain, Vg is the gate voltage, D is the thickness of the insulating layer, L is the channel length, W is the channel width, epsilon _r is the ratio of the gate insulating layer The dielectric constant and ε are the dielectric constant of vacuum (8.85 × 10 ⁻¹² F / m).
Further, the on / off ratio can be obtained from the ratio between the maximum value of Id and the minimum value of Id.

(CNT)
As the CNT, a single-layer CNT in which one carbon film (graphene sheet) is wound in a cylindrical shape, a two-layer CNT in which two graphene sheets are wound in a concentric circle, and a plurality of graphene sheets are concentric Any of multi-walled CNTs wound in a shape may be used, but single-walled CNTs are preferably used in order to obtain high semiconductor characteristics. CNT can be obtained by an arc discharge method, a chemical vapor deposition method (CVD method), a laser ablation method, or the like.

Further, it is more preferable that the CNT contains 80% by weight or more of the semiconductor CNT. More preferably, it contains 95% by weight or more of semiconducting CNTs. A known method can be used as a method for obtaining a semiconductor-type 80% by weight or more CNT. For example, a method of ultracentrifugation in the presence of a density gradient agent, a method of selectively attaching a specific compound to the surface of a semiconductor-type or metal-type CNT, and separating using a difference in solubility, a difference in electrical properties And a method of separation by electrophoresis or the like. Examples of the method for measuring the content of the semiconductor CNT include a method of calculating from the absorption area ratio of the visible-near infrared absorption spectrum and a method of calculating from the intensity ratio of the Raman spectrum.

In the present invention, the length of the CNT is preferably shorter than the distance between the first electrode and the second electrode in the applied semiconductor element or sensor. Specifically, although the average length of CNT depends on the channel length, it is preferably 2 μm or less, more preferably 1 μm or less. The average length of CNT refers to the average length of 20 CNTs picked up randomly. As a method for measuring the average CNT length, 20 CNTs are randomly picked up from images obtained by an atomic force microscope, a scanning electron microscope, a transmission electron microscope, or the like, and the average of their lengths is obtained. A method for obtaining the value is mentioned.

In general, commercially available CNTs are distributed in length and may contain CNTs that are longer than between the electrodes. Therefore, it is preferable to add a step of making the CNTs shorter than the distance between the electrodes. For example, a method of cutting into short fibers by acid treatment with nitric acid, sulfuric acid or the like, ultrasonic treatment, or freeze pulverization is effective. Further, it is more preferable to use separation by a filter in view of improving purity.

Further, the diameter of the CNT is not particularly limited, but is preferably 1 nm or more and 100 nm or less, and more preferably 50 nm or less.

In the present invention, it is preferable to provide a step of uniformly dispersing CNT in a solvent and filtering the dispersion with a filter. By obtaining CNT smaller than the filter pore diameter from the filtrate, CNT shorter than between the electrodes can be obtained efficiently. In this case, a membrane filter is preferably used as the filter. The pore diameter of the filter used for the filtration may be smaller than the channel length, and is preferably 0.5 μm or more and 10 μm or less.
Other methods for shortening CNT include acid treatment, freeze pulverization treatment, and the like.
The semiconductor layer of the present invention preferably contains carbon nanotubes having a high mobility and a large specific surface area.

(Conjugated polymer)
In the present invention, it is preferable that a conjugated polymer is attached to at least a part of the CNT surface. The conjugated polymer prevents an unexpected change in electrical characteristics from directly contacting the sample solution with the semiconductor component, and also plays a role of assisting electron transfer of the semiconductor component by the conjugated system.

Examples of the conjugated polymer include a polythiophene polymer, a polypyrrole polymer, a polyaniline polymer, a polyacetylene polymer, a poly-p-phenylene polymer, and a poly-p-phenylene vinylene polymer. Although it is mentioned, it is not specifically limited. As the polymer, those in which single monomer units are arranged are preferably used, but those obtained by block copolymerization or random copolymerization of different monomer units are also used. Further, graft-polymerized products can also be used.

In the present invention, among the above polymers, a polythiophene polymer that is easy to adhere to CNT and easily forms a complex with CNT is particularly preferable. The preferred molecular weight of the conjugated polymer is 800 or more and 100,000 or less in terms of number average molecular weight. Further, the polymer need not necessarily have a high molecular weight, and may be an oligomer composed of a linear conjugated system.

The conjugated polymer contains a side chain, and at least a part of the side chain is a hydroxyl group, a carboxy group, an amino group, a mercapto group, a sulfo group, a phosphonic acid group, an organic salt or an inorganic salt thereof, a formyl group. , Preferably containing at least one functional group selected from the group consisting of a maleimide group and a succinimide group, and particularly preferably containing at least one functional group selected from the group consisting of an amino group, a maleimide group and a succinimide group. . Thereby, it becomes easy to fix the biological substance which selectively interacts with the sensing target substance.

Of the above functional groups, the amino group, maleimide group, and succinimide group may or may not have a substituent. Examples of the substituent include an alkyl group, and this substituent may be further substituted. These functional groups may be bonded to form a ring. Furthermore, the other compound having the functional group may adhere to at least a part of the CNT surface.

The side chain in the present invention refers to a chain containing at least one carbon atom connected by substitution with an atom constituting the main chain of the conjugated polymer. The term “containing a functional group in a side chain” means that the functional group is included at the end of the side chain, or that the functional group is branched from the side chain and includes the functional group. A chain is a chain in which two or more atoms are connected in series. At this time, one of the atoms contained in the functional group can be included in the atoms constituting the molecular chain. Thus, for example, when a group represented by CH ₂ —COOH is linked to the main chain, this is a side chain containing a carboxy group.

This side chain preferably contains an alkylene group in at least a part of the chain. The alkylene group may be directly bonded to the atoms constituting the conjugated polymer that is the main chain, or may be bonded via an ether bond, an ester bond, or the like.

Here, the alkylene group includes, for example, a methylene group, an ethylene group, an n-propylene group, an isopropylene group, an n-butylene group, a sec-butylene group, a tert-butylene group, a cyclopropylene group, a cyclohexylene group, and a norbornylene group. A divalent saturated aliphatic hydrocarbon group such as, may or may not have a substituent. In the case of having a substituent, the additional substituent is not particularly limited, and examples thereof include an alkyl group, an alkoxy group such as a methoxy group and an ethoxy group, and these further have a substituent. May be. Moreover, although carbon number of an alkylene group is not specifically limited, 1 or more and 20 or less are preferable from the point of availability or cost, More preferably, it is 1 or more and 8 or less.

Specific examples of the conjugated polymer having the functional group in the side chain include the following structures. In addition, n in each structure shows the number of repetitions, and is the range of 2 or more and 1000 or less. Further, the conjugated polymer may be a single polymer of each structure or a copolymer. Moreover, the copolymer of each structure and the structure which does not have a side chain may be sufficient.

The conjugated polymer used in the present invention can be synthesized by a known method. To synthesize monomers, for example, as a method of linking thiophene and a thiophene derivative in which an alkyl group having a carboxy group at the end is introduced into the side chain, a halogenated thiophene derivative and thiophene boronic acid or thiophene boronic acid ester are used as palladium catalysts. And a method of coupling a halogenated thiophene derivative and a thiophene Grignard reagent under a nickel or palladium catalyst. In addition, when thiophene is linked to another unit having a functional group introduced, it can be coupled in the same manner using a halogenated unit. In addition, a conjugated polymer can be obtained by introducing a polymerizable substituent at the terminal of the monomer thus obtained and allowing the polymerization to proceed under a palladium catalyst or a nickel catalyst.

The conjugated polymer used in the present invention preferably removes impurities such as raw materials and by-products used in the synthesis process. For example, silica gel columnography, Soxhlet extraction, filtration, ion exchange, chelation, and the like can be used. Two or more of these methods may be combined.

When a CNT composite is used as a semiconductor component in the semiconductor layer of the present invention, by attaching an organic substance to at least a part of the CNT surface, the CNT can be put into a solution without impairing the high electrical properties possessed by the CNT. It becomes possible to disperse uniformly. Further, a uniformly dispersed CNT film can be formed from a solution in which CNTs are uniformly dispersed by a coating method. Thereby, a high semiconductor characteristic is realizable.

For example, (I) a method of adding and mixing CNT in a molten organic material, and (II) a method of dissolving an organic material in a solvent and adding and mixing CNT therein. , (III) CNTs are predispersed in advance with ultrasonic waves, etc., and organic substances are added and mixed therewith, and (IV) organic substances and CNTs are placed in a solvent, and this mixed system is irradiated with ultrasonic waves. The method of mixing etc. is mentioned. In the present invention, any method may be used, and any method may be combined.

The organic substance is not particularly limited, but specifically, polyvinyl alcohol, celluloses such as carboxymethyl cellulose, polyalkylene glycols such as polyethylene glycol, and acrylic resins such as polyhydroxymethyl methacrylate, poly Examples include conjugated polymers such as -3-hexylthiophene, polycyclic aromatic compounds such as anthracene derivatives and pyrene derivatives, and long-chain alkyl organic salts such as sodium dodecyl sulfate and sodium cholate.

From the viewpoint of interaction with CNT, those having a hydrophobic group such as an alkyl group and an aromatic hydrocarbon group or those having a conjugated structure are preferred, and a conjugated polymer is particularly preferred. If it is a conjugated polymer, the effect of uniformly dispersing CNT in the solution and the effect of high semiconductor properties are further improved without impairing the high electrical properties possessed by the CNTs.

(Protective agent)
When the sensor containing the semiconductor element of the present invention is used, the semiconductor layer may be treated with a reagent (referred to as “protective agent”) for preventing the approach and adsorption of substances other than the detection target substance. As a result, the detection target substance can be selectively detected. The protective agent may be physically adsorbed on the semiconductor layer, or may be introduced somewhere in the semiconductor layer through a bond.

Examples of the method for attaching the protective agent to the semiconductor layer include: (I) a method in which a semiconductor component is preliminarily dispersed with ultrasonic waves or the like, and a protective agent is added thereto and mixed; (II) a protective agent in a solvent. And a semiconductor component, and a method of mixing the mixed system by irradiating ultrasonic waves to this mixed system, (III) a method of immersing the semiconductor component coated on the substrate in a molten protective agent, and (IV) a protective agent in a solvent And a method of immersing the semiconductor component coated on the substrate in the solution. In the present invention, any method may be used, and any method may be combined. From the viewpoint of detection sensitivity, a method of attaching a protective agent to a semiconductor component using a solid-liquid reaction such as (III) or (IV) is preferable.

The conjugated polymer and the protective agent may be the same compound or different compounds. From the viewpoint of detection sensitivity, different compounds are preferable. Examples of the protective agent include proteins such as bovine serum albumin, casein, and skim milk, celluloses such as carboxymethyl cellulose, polyalkylene glycols such as polyethylene glycol, ethanolamine, and polyvinyl alcohol.
The order of attaching the conjugated polymer and the protective agent to the semiconductor component is not particularly limited, but it is preferable to attach the protective agent after attaching the conjugated polymer.

(Another biological substance)
In the sensor of the present invention, another biological substance that selectively interacts with the biological substance that is a sensing target substance is preferably fixed to the semiconductor layer. Hereinafter, another biological substance is referred to as “receptor”.

The receptor is not particularly limited as long as it can selectively interact with the sensing target substance, and any substance can be used. Specifically, enzyme, antigen, antibody, hapten, hapten antibody, peptide, oligopeptide, polypeptide (protein), hormone, nucleic acid, oligonucleotide, biotin, biotinylated protein, avidin, streptavidin, sugar, oligosaccharide, And saccharides such as polysaccharides, low-molecular compounds, high-molecular compounds, inorganic substances and complexes thereof, viruses, bacteria, cells, biological tissues, and substances constituting them.

Of these, antibodies, aptamers, enzymes, low molecular compounds, proteins, and oligonucleotides are preferable, low molecular compounds, antibodies, aptamers, and enzymes are more preferable, and biotin, antibodies, and aptamers are particularly preferable.

Examples of the low molecular weight compound include compounds having a molecular weight of about 100 to 1000, and biotin, pyrenebutanoic acid succinimide ester, pyrenebutanoic acid maleimide ester, and the like.

Examples of antibodies include anti-PSA, anti-hCG, anti-IgE, anti-BNP, anti-NT-proBNP, anti-AFP, anti-CK-MB, anti-PIVKA II, anti-CA15-3, anti-CA15-3, -CYFRA, anti-HIV, anti-troponin T, anti-procalcitonin, anti-HbA1c, anti-apolipoprotein, and anti-C reactive protein (CRP). Among them, the IgG type is preferable, and an antibody having only a variable site (Fab) fragment is particularly preferable.

Examples of aptamers include oligonucleotide aptamers and peptide aptamers. Specific examples include IgE aptamer, PSA aptamer, and thrombin aptamer. Examples of the enzyme include glucose oxidase and peroxidase. Of these, biotin, anti-IgE, anti-PSA, and IgE aptamer are more preferable.

The method for immobilizing the receptor to the semiconductor layer is not particularly limited, but biologically related substances other than glucose and functional groups contained in the semiconductor layer, that is, hydroxyl group, carboxy group, amino group, mercapto group, sulfo group. It is preferable to utilize a reaction or interaction with at least one functional group selected from the group consisting of a group, a phosphonic acid group, an organic salt or an inorganic salt thereof, a formyl group, a maleimide group and a succinimide group.

From the viewpoint of immobilization strength, it is preferable to use a reaction or interaction between a biological substance other than glucose and a functional group contained in the semiconductor layer. For example, when a bio-related substance other than glucose contains an amino group, examples thereof include a carboxy group, an aldehyde group, and a succinimide group. In the case of a thiol group, a maleimide group and the like can be mentioned.

Among the above, the carboxy group and the amino group can easily utilize the reaction or interaction with the receptor, making it easy to fix the receptor to the semiconductor layer. Therefore, it is preferable that the functional group contained in at least a part of the CNT is a carboxy group and an amino group.

Specific examples of the reaction or interaction include chemical bond, hydrogen bond, ionic bond, coordination bond, electrostatic force, van der Waals force, etc., but are not particularly limited. The type of functional group and the chemical structure of the receptor Appropriate selection may be made according to the situation. Further, if necessary, a part of the functional group and / or receptor may be converted into another suitable functional group and then fixed. Further, a linker such as terephthalic acid may be used between the functional group and the receptor.

The length of the functional group contained in the CNT is preferably short so that the receptor can be held close to the semiconductor component. This is because the sensing target substance is captured by the receptor closer to the semiconductor layer, and the detection signal becomes larger. More specifically, the length of the functional group contained in the CNT is preferably from 0.1 nm to 5 nm, more preferably from 0.15 nm to 3.1 nm, and particularly preferably from 0.3 nm to 1.6 nm.

In order to ensure the mobility of the functional group contained in the CNT in the sample solution, it is preferable that the functional group contained in the CNT contains a bond having a high affinity with water. Examples of such a bond include an ether bond, a thioether bond, an ester bond, an amide bond, a thioester bond, a dithioester bond, an acid anhydride bond, and an imide bond. Among these, an ether bond, an ester bond, an amide bond, and an imide bond are particularly preferable from the viewpoints of stability and affinity with water. Moreover, the ring structure may exist in the functional group which CNT contains.

The fixing process is not particularly limited, but a solution containing the receptor is dropped on the semiconductor layer containing CNTs, and after fixing the receptor while applying heating, cooling, vibration, etc. as necessary, excess components are removed. The process etc. which are removed by washing | cleaning or drying are mentioned.

In the second detection element of the sensor of the present invention, the combination of the functional group / receptor / sensing target substance contained in CNT is, for example, carboxy group / T-PSA-mAb (monoclonal antibody for prostate specific antigen) / PSA. (Prostate specific antigen), carboxy group / anti-hCG-mAb (anti-human chorionic gonadotropin antibody) / hCG (human chorionic gonadotropin), carboxy group / artificial oligonucleotide / IgE (immunoglobulin E), carboxy group / diisopropylcarbodiimide / IgE, carboxy group / amino group terminal RNA / HIV-1 (human immunodeficiency virus), carboxy group / anti-natriuretic peptide antibody / BNP (brain natriuretic peptide), carboxy group / anti-AFP polyclonal antibody (human tissue immunostaining for Body) / α-fetoprotein, carboxy group / anti-troponin T (anti-troponin T antibody) / troponin T, carboxy group / anti-CK-MB (anti-creatinine kinase MB antibody) / CK-MB (creatinine kinase MB), carboxy group / Anti-PIVKA-II (anti-protein induced by vitamin K absence or antagonist (PIVKA) -II antibody) / PIVKA-II, carboxy group / anti-CA15-3 (breast cancer C tumor marker) antibody / CA15-3, carboxy group / Anti-CEA (anti-carcinoembryonic antigen antibody) / CEA (carcinoembryonic antigen), anti-CYFRA (anti-cytokeratin 19 fragment antibody) / CYFRA (cytokeratin 19 fragment), Ruboxy group / anti-p53 (anti-p53 protein antibody) / p53 (p53 protein), carboxy group / anti-human hemoglobin monoclonal antibody / hemoglobin, carboxy group / anti-mouse hemoglobin A1c monoclonal antibody / HbA1c, carboxy group / anti-human albumin antibody / Albumin, carboxy group / anti-glycalbumin antibody / glycoalbumin, amino group / RNA / HIV-1, amino group / biotin / avidin, mercapto group / T-PSA-mAb / PSA, mercapto group / hCG-mAb / hCG, sulfo Group / T-PSA-mAb / PSA, sulfo group / hCG-mAb / hCG, phosphonic acid group / T-PSA-mAb / PSA, phosphonic acid group / hCG-mAb / hCG, aldehyde group / oligonucleotide / nucleic acid, aldehyde Group / anti-AFP polyclonal antibody (human tissue immunostaining antibody) / alpha-fetoprotein, a maleimide group / cysteine /, and ester / streptavidin / biotin and the like.

In addition, when the receptor contains a functional group, a combination of a compound containing the functional group (= another biological substance) / sensing target substance can be preferably used. Specifically, IgE aptamer / IgE, Combinations of biotin / avidin, stop preavidin / biotin, natriuretic peptide receptor / BNP (brain natriuretic peptide) and the like can be mentioned. Among these, in the sensor of the present invention, target substances having great significance to be detected together with glucose are hemoglobin, HbA1c, and glycoalbumin, and it is particularly preferable to use a sensor for detecting these.

(Sensor)
As described above, the sensor of the present invention includes at least a first detection element that detects glucose in a biological fluid and a second detection element that detects a biological substance other than glucose, and the second detection element. It is a small sensor that can be measured on the same substrate including a semiconductor element. Furthermore, even when the third detection element includes a semiconductor element, it is a small sensor that can be measured on the same substrate. In particular, a sensor part containing a semiconductor element will be described in detail. The sensor part includes a substrate, a first electrode, a second electrode, and a semiconductor layer, and the first electrode, the second electrode, and the semiconductor layer are formed on the substrate. The semiconductor layer contains a semiconductor element disposed between the first electrode and the second electrode. Furthermore, it is preferable that the semiconductor layer has another biological substance that selectively interacts with a biological substance other than glucose.

A sensor including a semiconductor element formed as shown in FIG. 2 includes a first electrode 103 and a second electrode 104 when a substance to be detected or a solution, gas or solid containing the substance to be detected is disposed in the vicinity of the semiconductor layer 111. The current value or electric resistance value flowing between the two changes. By measuring the change, the detection target substance can be detected.

In the sensor containing the semiconductor element of FIG. 3, the value of the current flowing through the semiconductor layer 111 can be controlled by the voltage of the third electrode 105. Accordingly, when the value of the current flowing between the first electrode 103 and the second electrode 104 when the voltage of the third electrode 105 is changed, a two-dimensional graph (IV graph) is obtained.

The detection target substance may be detected using part or all of the characteristic values, or the detection target substance may be detected using the ratio between the maximum current and the minimum current, that is, the on / off ratio. Furthermore, known electrical characteristics obtained from a semiconductor element such as resistance value, threshold voltage change, impedance, mutual conductance, and capacitance may be used.

The detection target substance may be used alone, or may be mixed with other substances or solvents. A substance to be detected or a solution, gas or solid containing the substance to be detected is disposed in the vicinity of the semiconductor layer 111. As described above, the electrical characteristics of the semiconductor layer 111 change due to the interaction between the semiconductor layer 111 and the substance to be detected, and this is detected as a change in any one of the electrical signals described above.

It is preferable that the sensor of the present invention further includes a covering member that covers at least a part of the substrate on the substrate. For example, as a modification of the configuration shown in FIG. 4, it is preferable to include an upper substrate 20 that forms an internal space between the substrate 10 and the substrate 10 as shown in FIGS. 5A and 5B. . A broken line in the upper substrate 20 in FIG. 5A indicates a boundary between the upper substrate 20 and the internal space. FIG. 5B is a cross-sectional view taken along line CC ′ of FIG. 5A and shows an internal space 108 between the substrate 10 and the upper substrate 20.

As another modification of the configuration shown in FIG. 4, it is preferable to include a covering member 21 that forms a space 108 surrounding the semiconductor layer 111 on the substrate 10 as shown in FIGS. 6A and 6B. . FIG. 6B is a cross-sectional view taken along line DD ′ of FIG. 6A. Thereby, it becomes possible to efficiently contact the semiconductor layer 111 and the liquid containing the detection target substance.

As another embodiment of the sensor of the present invention, it is preferable that the above-mentioned upper substrate or covering member is provided on a substrate, and a third electrode is provided on the surface of the upper substrate or covering member facing the semiconductor layer. That is, the substrate includes a substrate, a first electrode, a second electrode, and a semiconductor layer formed between the first electrode and the second electrode, and further includes an upper substrate or a covering member on the substrate, It is preferable that a third electrode is provided on a surface of the substrate or covering member that faces the semiconductor layer, and the semiconductor layer has another biological substance that selectively interacts with a biological substance other than glucose. In the space between the first electrode, the second electrode, and the semiconductor layer and the third electrode, either a gas layer, a liquid layer, a solid layer, or a combination thereof may exist, Good.

FIG. 7 is a schematic cross-sectional view showing an example of a sensor portion containing the semiconductor element of the present invention. In the sensor of FIG. 7, the first electrode 103 and the second electrode 104 are formed on the substrate 10, the semiconductor layer 111 is disposed between the first electrode 103 and the second electrode 104, and the upper substrate 20 is on the substrate 10. Are arranged on the same side as the first electrode 103, the second electrode 104, and the semiconductor layer 111, and the third electrode 105 is arranged on the upper substrate 20. The arrangement of the third electrode 105 on the upper substrate 20 is not limited to the position immediately above the semiconductor layer, but may be an oblique upper side. Further, the upper substrate 20 is not limited to the upper surface portion when viewed from the semiconductor layer, and may be disposed on the side surface. The third electrode 105 is not limited to being disposed on the upper substrate 20, and may be disposed on the substrate 10.

Examples of the material used for the upper substrate 20 or the covering member include inorganic materials such as silicon wafer, glass, and alumina sintered body, polyimide, polyester, polycarbonate, polysulfone, polyethersulfone, polyethylene, polyphenylene sulfide, and polyparaxylene. However, it is not limited to these. These materials may be used alone, or a plurality of materials may be laminated or mixed.

(Sensor manufacturing method)
The manufacturing method of the sensor containing the 1st detection element 401 shown in FIG. 1 and the 2nd detection element 402 is shown. The sensor manufacturing method includes a step of forming a semiconductor layer by applying and drying a semiconductor component on a substrate. The manufacturing method is not limited to the following.

In particular, when describing details of a sensor portion containing a semiconductor element, the first electrode 103 and the second electrode 104 are formed on the substrate 10. Examples of the forming method include known methods such as metal deposition, spin coating method, blade coating method, slit die coating method, screen printing method, bar coater method, mold method, printing transfer method, immersion pulling method, and ink jet method. . It is also possible to directly form a pattern using a mask or the like, or to apply a resist on a substrate, expose and develop the resist film in a desired pattern, and then pattern the gate electrode by etching. . Here, the connection portion 102 and the wiring 107 may be formed by the same formation method as the formation method of the first electrode and the second electrode, or each may be formed in a lump.

The connection part 102, the wiring 107 and the first electrode 103 are electrically connected by the second electrode 104, the wiring 107 and the connection part 102 through the semiconductor layer 111. Although not shown, a power source and a detection unit are connected to the connection unit 102. Using this connection, a voltage can be applied between the electrodes from the power source, and an electrical signal generated in the semiconductor layer can be taken out from the connection portion.

Next, the semiconductor layer 111 is formed. A manufacturing method, comprising: attaching a semiconductor layer to a compound containing a linking group on the semiconductor layer; and forming a bond between the linking group and another bio-related substance that selectively interacts with the bio-related substance. Is preferred.

As a method for attaching a compound containing a linking group to the semiconductor layer, for example, a method of vapor-depositing a compound containing a linking group in a vacuum, or by immersing the semiconductor layer in a solution in which the compound containing the linking group is dissolved. Examples thereof include a method, a method of applying a compound containing a linking group to the semiconductor layer, and a method of applying a solution in which the compound containing the linking group is dissolved in the semiconductor layer.

Examples of the step of forming a bond between another biological substance that selectively interacts with the biological substance and the linking group include, for example, a method of causing another biological substance to collide with the semiconductor layer in a vacuum to cause a reaction. And a method in which the semiconductor layer is immersed in a solution in which another biological substance is dissolved, and a method in which a solution in which another biological substance is dissolved is applied to the semiconductor layer.

The formation of the semiconductor layer and the fixation of another biological substance may be performed separately or collectively. For example, a method of forming a semiconductor layer using a semiconductor component in which another biological substance is bonded or attached in advance through a linking group can be used.

(Sensor shape)
As the shape of the sensor, the shape shown in FIG. 1 of the present invention is preferable. Specific shapes include a shape like a sensor chip used in a blood glucose level sensor and a portable cassette type. These shapes are preferable because the size of the main body having the power source to be connected and the analysis algorithm can be reduced.

In a device using conventional light, a light source is required, and the device is enlarged because a unit for diffusing heat generated by the light source is required, but according to the present invention, a light source is not used. The size can be reduced as compared with the conventional apparatus.

(Sensor that detects hemoglobin, glycated hemoglobin, glycated albumin, etc.)
Another embodiment of the present invention includes a sensor having a detection element that detects at least one selected from the group consisting of hemoglobin, glycated hemoglobin, and glycated albumin, and the detection element includes a semiconductor element. This is an application of the above-described second detection element portion as a single sensor. Therefore, the structure of the sensor, the material used for the sensor, the action of the sensor, and the like are the same as described above.

In the present embodiment, the functional group / receptor / sensing target substance combination contained in the CNT complex in the semiconductor element is, for example, carboxy group / anti-human hemoglobin monoclonal antibody / hemoglobin, carboxy group, carboxy group / anti-antibody. Examples include mouse hemoglobin A1c monoclonal antibody / glycohemoglobin, carboxy group / anti-human albumin antibody / albumin, and carboxy group / anti-glycoalbumin antibody / glycoalbumin.

Hereinafter, specific examples in the case of detecting glucose, glycated hemoglobin or glycated albumin, and hemoglobin in blood using the sensor of the present invention will be described. The following examples are examples of the present invention, and the types of materials and hemolytic agents used for the semiconductor layer can be appropriately changed as described in this specification. In addition, various modifications can be made as described in the present specification depending on the types of biological substances detected by the second detection element and the third detection element.

First, blood as a measurement sample is injected into the sensor from the injection port 301. The injected blood flows through the path 302 and branches in the middle of the first detection element direction and the second detection element direction.
When the blood flowing toward the first detection element reaches the reaction layer 110, glucose in the blood reacts with glucose degrading enzyme in the reaction layer 110. Utilizing this, the glucose concentration is measured.

The blood flowing toward the second detection element is mixed with a hemolytic agent in a space 303 in the middle. Thereby, HbA1c is taken out from the red blood cells.
In the second detection element, the semiconductor layer 111 contains a CNT complex, and an anti-mouse hemoglobin A1c monoclonal antibody that selectively interacts with HbA1c is immobilized on the CNT complex. When HbA1c extracted from the red blood cells in the space 303 reaches the semiconductor layer 111, the value of the current flowing through the semiconductor layer 111 changes due to the interaction with the anti-mouse hemoglobin A1c monoclonal antibody. Using this, the HbA1c concentration is measured.

The blood flowing in the direction of the third detection element is mixed with the hemolytic agent in the space 303 in the middle. Thereby, hemoglobin is taken out from the red blood cells.
In the third detection element, the semiconductor layer 201 contains a CNT complex, and an anti-hemoglobin antibody that selectively interacts with hemoglobin is immobilized on the CNT complex. When hemoglobin extracted from the red blood cells in the space 303 reaches the semiconductor layer 201, the value of current flowing through the semiconductor layer 201 changes due to the interaction with the anti-hemoglobin antibody. Using this, the hemoglobin concentration is measured.

Hereinafter, the present invention will be described more specifically based on examples. In addition, this invention is not limited to the following Example. The used CNTs are as follows.
SWCNT: Meijo Nano Carbon Co., Ltd., single-walled CNT, semiconducting CNT purity> 95%
DWCNT: Two-layer CNT manufactured by Toray Industries, Inc.
MWCNT: Multi-layer CNT, manufactured by Meijo Nanocarbon

Moreover, it shows below about what used the abbreviation among the compounds used for the Example and the comparative example.
P3HT: poly-3-hexylthiophene PBS: phosphate buffered saline BSA: bovine serum albumin IgE: immunoglobulin E
FBS: fetal bovine serum NT-proBNP: human brain natriuretic peptide precursor N-terminal fragment PBSE: 1-pyrenebutanoic acid-N-hydroxysuccinimide ester PET: polyethylene terephthalate PEN: polyethylene naphthalate o-DCB: o-dichlorobenzene In Examples 1 to 13, the sensor shown in FIG. 1 was produced, and in Examples 14 to 27, the sensor shown in FIG. 9 was produced and used for evaluation. In Examples 1 to 27, the first detection element was produced in the same manner as the production method of the conventional blood glucose level sensor, and it was confirmed that it could be detected.

Example 1
(1) Preparation of semiconductor solution Add 1.5 mg of CNT and 1.5 mg of P3HT into 15 mL of chloroform, and use an ultrasonic homogenizer (VCX-500, manufactured by Tokyo Rika Kikai Co., Ltd.) while cooling with ice at an output of 250 W. The mixture was ultrasonically stirred for 30 minutes to obtain CNT dispersion A (CNT complex concentration of 0.1 g / l with respect to the solvent).
Next, a semiconductor solution for forming a semiconductor layer was prepared. The CNT dispersion A was filtered using a membrane filter (pore size: 10 μm, diameter: 25 mm, Omnipore membrane manufactured by Millipore), and then a membrane filter (pore size: 3 μm, diameter: 25 mm, Omnipore membrane manufactured by Millipore) was used. And filtered. 45 mL of o-DCB was added to 5 mL of the obtained filtrate to obtain a semiconductor solution A (CNT complex concentration of 0.01 g / l with respect to the solvent).

(2) Preparation of polymer solution for insulating layer 61.29 g (0.45 mol) of methyltrimethoxysilane, 12.31 g (0.05 mol) of β- (3,4-epoxycyclohexyl) ethyltrimethoxysilane, and phenyl 99.15 g (0.5 mol) of trimethoxysilane was dissolved in 203.36 g of propylene glycol monobutyl ether (boiling point 170 ° C.), and 54.90 g of water and 0.864 g of phosphoric acid were added thereto with stirring. The obtained solution was heated at a bath temperature of 105 ° C. for 2 hours, the internal temperature was raised to 90 ° C., and a component mainly composed of methanol produced as a by-product was distilled off. Next, the bath was heated at 130 ° C. for 2.0 hours, the internal temperature was raised to 118 ° C., and a component mainly composed of water and propylene glycol monobutyl ether was distilled off, and then cooled to room temperature, and the solid content concentration was 26.0. A weight percent polymer solution A was obtained.
50 g of the obtained polymer solution A was weighed, mixed with 16.6 g of propylene glycol monobutyl ether (boiling point 170 ° C.), stirred at room temperature for 2 hours, and polymer solution B (solid content concentration 19.5 wt%) was obtained. Obtained.

(3) Fabrication of Semiconductor Element The semiconductor element shown in FIG. 3 was fabricated. On the glass substrate 10 (film thickness 0.7 mm), gold was vacuum-deposited by 50 nm through a mask by a resistance heating method, and the first electrode 103 and the second electrode 104 were formed. The width (channel width) of the first electrode and the second electrode was 400 μm, and the distance (channel length) between the first electrode and the second electrode was 60 μm.
A semiconductor layer 111 is formed by dropping 400 pl of the semiconductor solution A produced by the method described in (1) above on the organic film on which the electrode is formed using an inkjet apparatus (manufactured by Cluster Technology Co., Ltd.), and a hot plate The semiconductor device A was obtained by performing a heat treatment at 120 ° C. for 30 minutes under a nitrogen stream.
Next, the semiconductor layer was immersed overnight at 4 ° C. in a solution of Anti-HbA1c (manufactured by Funakoshi) at 0.01 ug / mL with 0.01 M PBS. Thereafter, the semiconductor layer was thoroughly rinsed with 0.01M PBS. Next, BSA (manufactured by Wako Pure Chemical Industries, Ltd.) 5.0 mg in 0.01 M PBS 5.0 mL was immersed for 2 hours. Thereafter, the semiconductor layer was sufficiently rinsed with 0.01 M PBS to obtain a semiconductor element in which the semiconductor layer was modified with Anti-HbA1c, which is a biological substance that selectively interacts with the sensing target substance, and BSA, which is a protective agent.

(4) Evaluation as sensor The semiconductor layer of the produced semiconductor element was immersed in 100 μl of 0.01M PBS, and the value of the current flowing between the first electrode and the second electrode was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The measurement was performed at a voltage between the first electrode and the second electrode (Vsd) = − 0.2V and a voltage between the first electrode and the third electrode (Vg) = − 0.7V. 60 μm after the start of measurement, 20 μl of 5 μg / mL BSA-0.01M PBS solution, 75 μm after 5 μg / mL Avidin (manufactured by Wako Pure Chemical Industries, Ltd.) — 0.01 μM PBS solution 20 μl, 90 μm after 5 μg / mL HbA1c (Funakoshi) —0.01 μM PBS solution 20 μl was added to 0.01 M PBS soaked in a semiconductor layer. The results are shown in Table 1. Only when HbA1c was added, the current value increased by 5.0% from the current value before the addition.

Example 2
(1) Production of Semiconductor Element Semiconductor element A was produced in the same manner as in Example 1.
Next, the semiconductor layer was immersed in 1.0 mL of 1-pyrenebutanoic acid-N-hydroxysuccinimide ester (anaspec Co., Ltd., PBSE) in 6.0 mL of acetonitrile (Wako Pure Chemical Industries, Ltd.) for 1 hour. . Thereafter, the semiconductor layer was sufficiently rinsed with acetonitrile and methanol (manufactured by Wako Pure Chemical Industries, Ltd.). The semiconductor layer was immersed overnight at 4 ° C. in a solution of Anti-HbA1c (manufactured by Funakoshi) made 100 ug / mL with 0.01 M PBS. Thereafter, the semiconductor layer was thoroughly rinsed with 0.01M PBS. Next, BSA (manufactured by Wako Pure Chemical Industries, Ltd.) 5.0 mg in 0.01 M PBS 5.0 mL was immersed for 30 minutes. Thereafter, the semiconductor layer was sufficiently rinsed with 0.01 M PBS to obtain a semiconductor element in which the semiconductor layer was modified with Anti-HbA1c, which is a biological substance that selectively interacts with the sensing target substance, and BSA, which is a protective agent.

(2) Evaluation as a sensor In order to evaluate the semiconductor element produced above as a sensor, the measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, a current value increase of 4.8% was observed from the current value before the addition.

Example 3
(1) Fabrication of Semiconductor Element The semiconductor element shown in FIG. 3 was fabricated. A glass substrate 10 (film thickness 0.7 mm) was subjected to ultraviolet ozone treatment (photo surface processor, PL30-200, manufactured by SEN LIGHTTS CORP.) For 30 minutes, and a polyethylene glycol chain-containing silane coupling agent (SIH6188, manufactured by Gelest) It was immersed in a 10 wt% ethanol solution for 1 hour. After washing with ethanol for 30 seconds, the organic film 106 was formed by drying at 120 ° C. for 30 minutes. The first electrode 103 and the second electrode 104 were formed on the organic film by mask vapor deposition so that the film thickness was 50 nm. The width (channel width) of the first electrode and the electric two electrode was 400 μm, and the distance (channel length) between the first electrode and the electric two electrode was 60 μm.
A semiconductor layer 4 is formed by dropping 400 pl of the semiconductor solution A produced by the method described in (1) above on the organic film on which the electrode is formed using an inkjet apparatus (manufactured by Cluster Technology Co., Ltd.), and a hot plate A heat treatment was performed at 150 ° C. for 30 minutes under a nitrogen stream to obtain a semiconductor element B.
Next, the semiconductor layer was immersed in a 1.0 mL solution of 6.0 mg of 1-pyrenebutanoic acid-N-hydroxysuccinimide ester (manufactured by Anaspec Co., Ltd.) in acetonitrile (manufactured by Wako Pure Chemical Industries, Ltd.) for 1 hour. Thereafter, the semiconductor layer was sufficiently rinsed with acetonitrile and methanol (manufactured by Wako Pure Chemical Industries, Ltd.). The semiconductor layer was immersed overnight at 4 ° C. in a solution of Anti-HbA1c (manufactured by Funakoshi) made 100 ug / mL with 0.01 M PBS. Thereafter, the semiconductor layer was thoroughly rinsed with 0.01M PBS. Next, BSA (manufactured by Wako Pure Chemical Industries, Ltd.) 5.0 mg in 0.01 M PBS 5.0 mL was immersed for 30 minutes. Thereafter, the semiconductor layer was sufficiently rinsed with 0.01 M PBS to obtain a semiconductor element in which the semiconductor layer was modified with Anti-HbA1c, which is a biological substance that selectively interacts with the sensing target substance, and BSA, which is a protective agent.

(2) Evaluation as a sensor In order to evaluate the semiconductor element produced above as a sensor, the measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, the current value increased by 6.0% from the current value before the addition.

Example 4
(1) Fabrication of semiconductor device A semiconductor layer was formed in the same manner as in Example 3 except that Anti-HbA1c fragment antibody (Fab) was used instead of Anti-HbA1c.
(2) Evaluation as a sensor In order to evaluate the semiconductor element produced above as a sensor, the measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, a current value increase of 6.5% was observed from the current value before the addition.

Example 5
(1) Fabrication of semiconductor element A semiconductor element was obtained by forming a semiconductor layer in the same manner as in Example 4 except that single-walled carbon nanotubes (SWCNT) containing 90% by weight of a metal mold were used instead of gold. .
(2) Evaluation as a sensor In order to evaluate the semiconductor element produced above as a sensor, the measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, a current value increase of 5.8% was observed from the current value before the addition.

Example 6
(1) Production of Semiconductor Element A semiconductor element was obtained by forming a semiconductor layer in the same manner as in Example 4 except that double-walled carbon nanotubes (DWCNT) were used instead of gold.
(2) Evaluation as a sensor In order to evaluate the semiconductor element produced above as a sensor, the measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, the current value increased by 6.0% from the current value before the addition.

Example 7
(1) Production of Semiconductor Element A semiconductor layer was formed in the same manner as in Example 4 except that multi-walled carbon nanotubes (MWCNT) were used instead of gold to obtain a semiconductor element.
(2) Evaluation as a sensor In order to evaluate the semiconductor element produced above as a sensor, the measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, a current value increase of 5.7% was observed from the current value before the addition.

Example 8
(1) Preparation of semiconductor solution Semiconductor solution B (CNT complex concentration 0.01 g / l with respect to the solvent) was carried out in the same manner as in Example 1 except that the compound represented by formula (69) was used instead of P3HT. It was.
(2) Fabrication of Semiconductor Element A semiconductor layer was formed in the same manner as in Example 4 except that the semiconductor solution B was used instead of the semiconductor solution A to obtain a semiconductor element.
(3) Evaluation as sensor In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, a current value increase of 6.6% was observed from the current value before the addition.

Example 9
(1) Preparation of semiconductor solution Semiconductor solution C (CNT complex concentration 0.01 g / l with respect to the solvent) was carried out in the same manner as in Example 1 except that the compound represented by formula (71) was used instead of P3HT. It was.
(2) Production of Semiconductor Element A semiconductor layer was formed in the same manner as in Example 4 except that the semiconductor solution C was used in place of the semiconductor solution A to obtain a semiconductor element.
(3) Evaluation as sensor In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 1. The results are shown in Table 1. Only when HbA1c was added, a current value increase of 6.9% was observed from the current value before the addition.

Example 10
(1) Fabrication of semiconductor device A semiconductor layer was formed in the same manner as in Example 9 except that anti-glycoalbumin was used instead of Anti-HbA1c.
(2) Evaluation as a sensor In order to evaluate the semiconductor element produced above as a sensor, the measurement was performed in the same manner as in Example 10. The results are shown in Table 1. Only when glycoalbumin was added, the current value increased by 8.1% from the current value before the addition.

Example 11
(1) Production of Semiconductor Element A semiconductor layer was formed in the same manner as in Example 10 except that PBSE was not used to obtain a semiconductor element.
(2) Evaluation as sensor The semiconductor layer of the manufactured semiconductor element was immersed in 100 μl of 0.01M PBS, and the value of the current flowing between the first electrode and the second electrode was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The measurement was performed at a voltage between the first electrode and the second electrode (Vsd) = − 0.2V and a voltage between the first electrode and the third electrode (Vg) = − 0.7V. 60 μm after the start of measurement, 20 μl of 5 μg / mL BSA-0.01M PBS solution, 75 μm after 5 μg / mL HbA1c (manufactured by Funakoshi) —0.01 μM PBS solution 20 μl, 90 min after 5 μg / mL glycoalbumin (manufactured by Funakoshi) — 20 μl of 0.01 M PBS solution was added to 0.01 M PBS soaked in the semiconductor layer. The results are shown in Table 1. Only when glycoalbumin was added, the current value increased by 9.8% from the current value before the addition.

Example 12
(1) Production of Semiconductor Device A semiconductor layer was formed in the same manner as in Example 11 except that anti-NT-proBNP was used instead of anti-glycoalbumin.
(2) Evaluation as sensor The semiconductor layer of the manufactured semiconductor element was immersed in 100 μl of 0.01M PBS, and the value of the current flowing between the first electrode and the second electrode was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The measurement was performed at a voltage between the first electrode and the second electrode (Vsd) = − 0.2V and a voltage between the first electrode and the third electrode (Vg) = − 0.7V. 60 μm after the start of measurement, 20 μl of 5 μg / mL BSA-0.01M PBS solution, 75 μm after 5 μg / mL HbA1c (manufactured by Funakoshi) -0.01 M PBS solution 20 μl, 90 min after 5 μg / mL NT-proBNP (manufactured by Funakoshi) -20 μl of 0.01 M PBS solution was added to 0.01 M PBS soaked in the semiconductor layer. The results are shown in Table 1. Only when NT-proBNP was added, a current value increase of 7.7% was observed from the current value before the addition.

Example 13
(1) Production of Semiconductor Device A semiconductor layer was formed in the same manner as in Example 10 except that anti-NT-proBNP was used instead of anti-glycoalbumin to obtain a semiconductor device.
(2) Evaluation as sensor In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 12. The results are shown in Table 1. Only when NT-proBNP was added, the current value increased by 7.5% from the current value before the addition.

Example 14
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
In addition, a semiconductor layer was formed in the same manner as in Example 10 except that anti-hemoglobin was used instead of anti-glycoalbumin, thereby producing a third detection element.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
The semiconductor layer of the manufactured semiconductor element was immersed in 100 μl of 0.01M PBS, and the value of current flowing between the first electrode and the second electrode was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The measurement was performed at a voltage between the first electrode and the second electrode (Vsd) = − 0.2V and a voltage between the first electrode and the third electrode (Vg) = − 0.7V. 60 μm after the start of measurement, 20 μl of 5 μg / mL BSA-0.01M PBS solution, 75 μm after 5 μg / mL HbA1c (manufactured by Funakoshi) -0.01 M PBS solution 20 μl, 90 μm after 5 μg / mL hemoglobin (manufactured by Funakoshi) -0 20 μl of 0.01 M PBS solution was added to 0.01 M PBS soaked in the semiconductor layer. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 3.2% was observed from the current value before the addition.

Example 15
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, a semiconductor layer was formed in the same manner as in Example 14 except that the width (channel width) of the first electrode and the electric two electrode was 400 μm and the interval (channel length) between the first electrode and the electric two electrode was 40 μm. A semiconductor element was manufactured to obtain a third detection element.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 14. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 4.0% was observed from the current value before the addition.

Example 16
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, a semiconductor layer was formed in the same manner as in Example 14 except that the width (channel width) of the first electrode and the electrode 2 was 400 μm and the interval (channel length) between the first electrode and the electrode 2 was 30 μm. A semiconductor element was manufactured to obtain a third detection element.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 14. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 4.8% was observed from the current value before the addition.

Example 17
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, a semiconductor layer was formed in the same manner as in Example 14 except that the width (channel width) of the first electrode and the electrode 2 was 400 μm and the interval (channel length) between the first electrode and the electrode 2 was 25 μm. A semiconductor element was manufactured to obtain a third detection element.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 14. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 5.1% was observed from the current value before the addition.

Example 18
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, a semiconductor layer was formed in the same manner as in Example 16 except that the substrate was made of glass, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 14. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 4.5% was observed from the current value before the addition.

Example 19
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
A semiconductor layer was formed in the same manner as in Example 16 except that the substrate was a PET film, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 14. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 4.7% was observed from the current value before the addition.

Example 20
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, a semiconductor layer was formed in the same manner as in Example 16 except that the substrate was a PEN film, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 14. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 4.6% was observed from the current value before the addition.

Example 21
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, in the same manner as in Example 16, a semiconductor layer was formed, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
The semiconductor layer of the produced semiconductor element was immersed in 100 μl of FBS (manufactured by BioWest), and the value of current flowing between the first electrode and the second electrode was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The measurement was performed at a voltage between the first electrode and the second electrode (Vsd) = − 0.2V and a voltage between the first electrode and the third electrode (Vg) = − 0.7V. 60 μm after the start of measurement, 20 μl of 5 μg / mL BSA-FBS solution, 75 μm after 5 μg / mL HbA1c (Funakoshi) -FBS solution 20 μl, 90 minutes later, 20 μl of healthy human blood were added to the FBS soaked in the semiconductor layer. The results are shown in Table 1. Only when blood was added to healthy people, a current value increase of 5.2% was observed from the current value before the addition, and blood hemoglobin was selectively detected.

Example 22
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, in the same manner as in Example 16, a semiconductor layer was formed, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as a sensor (second detection element)
The semiconductor layer of the produced semiconductor element was immersed in 100 μl of FBS (manufactured by BioWest), and the value of current flowing between the first electrode and the second electrode was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The measurement was performed at a voltage between the first electrode and the second electrode (Vsd) = − 0.2V and a voltage between the first electrode and the third electrode (Vg) = − 0.7V. 60 minutes after the start of the measurement, 20 μl of a 5 μg / mL BSA-FBS solution was added to FBS soaked with 20 μl of healthy human blood in a semiconductor layer 75 minutes later. The results are shown in Table 1. Only when a healthy person added blood, an increase in current value of 8.0% was observed from the current value before addition, and blood HbA1c was selectively detected.
(Third detection element)
The semiconductor layer of the produced semiconductor element was immersed in 100 μl of FBS (manufactured by BioWest), and the value of current flowing between the first electrode and the second electrode was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The measurement was performed at a voltage between the first electrode and the second electrode (Vsd) = − 0.2V and a voltage between the first electrode and the third electrode (Vg) = − 0.7V. 60 minutes after the start of the measurement, 20 μl of a 5 μg / mL BSA-FBS solution was added to FBS soaked with 20 μl of healthy human blood in a semiconductor layer 75 minutes later. The results are shown in Table 1. Only when blood was added to a healthy person, a current value increase of 5.3% was observed from the current value before the addition, and blood hemoglobin was selectively detected.

Example 23
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Also, implementation was performed except that anti-hemoglobin was used instead of Anti-HbA1c, the width of the first electrode and the electric two electrode (channel width) was 400 μm, and the distance between the first electrode and the electric two electrode (channel length) was 120 μm. In the same manner as in Example 14, a semiconductor layer was formed, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element)
In order to evaluate the semiconductor element produced above as a sensor, measurement was performed in the same manner as in Example 14. The results are shown in Table 1. Only when hemoglobin was added, a current value increase of 1.7% was observed from the current value before the addition.

Example 24
(1) Production of Semiconductor Element The second detection element was produced in the same manner as in Example 9.
Further, a semiconductor layer was formed in the same manner as in Example 14 except that the width (channel width) of the first electrode and the electrode 2 was 400 μm and the interval (channel length) between the first electrode and the electrode 2 was 600 μm. A semiconductor element was manufactured to obtain a third detection element.
(2) Evaluation as a sensor (second detection element)
The same results as in Example 9 were obtained.
(Third detection element) Measurement was carried out in the same manner as in Example 14 in order to evaluate the semiconductor element produced above as a sensor. With each addition of BSA, HbA1c, and hemoglobin, a current value increase of 0.3% was observed from the current value before the addition. The results are shown in Table 1.

Example 25
(1) Production of semiconductor element The first detection element was produced by the same production method as that of an existing blood glucose level sensor.
The second detection element was produced in the same manner as in Example 9.
Further, in the same manner as in Example 16, a semiconductor layer was formed, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as sensor The sensor shown in FIG. 9 was prepared, and 200 μl of FBS (manufactured by BioWest) was filled in the flow path, and the current value flowing between the electrodes of the second detection element and the third detection element Was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The voltage between the first and second electrodes (Vsd) of each of the second and third sensing elements (Vsd) = − 0.2V, and the voltage between the first and third electrodes (Vg) = − 0.7V. It was measured. Sixty minutes after the start of measurement, 20 μl of healthy human blood was added to FBS. The blood reached each detection element after hemolysis in the space in the flow path. The results are shown in Table 1. A change in current value derived from blood glucose level is obtained from the first detection element, and a current value increase of 8.4% is seen from the current value before addition from the second detection element, and blood HbA1c is selected. Detected. Furthermore, the current value increased by 5.5% from the current value before the third detection element, and blood hemoglobin was selectively detected.

Example 26
(1) Production of semiconductor element The first detection element was produced by the same production method as that of an existing blood glucose level sensor.
The second detection element was produced in the same manner as in Example 9.
Further, in the same manner as in Example 17, a semiconductor layer was formed, a semiconductor element was produced, and a third detection element was obtained.
(2) Evaluation as sensor The sensor shown in FIG. 9 was prepared, and 200 μl of FBS (manufactured by BioWest) was filled in the flow path, and the current value flowing between the electrodes of the second detection element and the third detection element Was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The voltage between the first and second electrodes (Vsd) of each of the second and third sensing elements (Vsd) = − 0.2V, and the voltage between the first and third electrodes (Vg) = − 0.7V. It was measured. Sixty minutes after the start of measurement, 20 μl of healthy human blood was added to FBS. The results are shown in Table 1.
A change in current value derived from blood sugar level is obtained from the first detection element, and a current value increase of 8.5% from the current value before addition is seen from the second detection element, and blood HbA1c is selected. Detected. Furthermore, the current value increased by 5.8% from the current value before the third detection element, and blood hemoglobin was selectively detected.

Example 27
(1) Production of semiconductor element The first detection element was produced by the same production method as that of an existing blood glucose level sensor.
The second detection element was produced in the same manner as in Example 9.
Further, a semiconductor layer was formed in the same manner as in Example 14 to produce a semiconductor element, thereby obtaining a third detection element.
(2) Evaluation as sensor The sensor shown in FIG. 9 was prepared, and 200 μl of FBS (manufactured by BioWest) was filled in the flow path, and the current value flowing between the electrodes of the second detection element and the third detection element Was measured. For the measurement, a semiconductor characteristic evaluation system 4200-SCS type (manufactured by Keithley Instruments Co., Ltd.) was used. The voltage between the first and second electrodes (Vsd) of each of the second and third sensing elements (Vsd) = − 0.2V, and the voltage between the first and third electrodes (Vg) = − 0.7V. It was measured. Sixty minutes after the start of measurement, 20 μl of healthy human blood was added to FBS. The results are shown in Table 1.
A change in current value derived from blood sugar level is obtained from the first detection element, and a current value increase of 8.5% from the current value before addition is seen from the second detection element, and blood HbA1c is selected. Detected. Further, the third detection element showed a 3.7% increase in the current value from the current value before the addition, and blood hemoglobin was selectively detected.

Comparative Example 1
HbA1c and hemoglobin were measured using a 7180 Hitachi automatic analyzer described in the examples of Japanese Patent Application Laid-Open No. 2015-165827 without using semiconductor elements for the second and third detection elements. As a result, since the analysis device is an optical instrument and the measures are large, it is difficult to perform electrical measurement using the same chip as glucose measurement.

Although the present invention has been described in detail using specific embodiments, it will be apparent to those skilled in the art that various modifications and variations can be made without departing from the spirit and scope of the invention. This application is based on a Japanese patent application filed on November 9, 2015 (Japanese Patent Application No. 2015-219207), which is incorporated by reference in its entirety.

The sensor using the present invention can be applied to various types of sensing such as chemical analysis, physical analysis, and biological analysis, and is particularly suitably used as a medical sensor or biosensor.

10 substrate 20 upper substrate 21 covering member 101 electrode 102 connecting portion 103 first electrode 104 second electrode 105 third electrode 106 insulating layer 107 wiring 108 internal space 110 reaction layer 111 semiconductor layer 112 reaction layer 121 electrode 122 first electrode 123 first 2 electrode 200 semiconductor element 201 semiconductor layer 202 semiconductor element 301 inlet 302 path 303 space 304 recess 401 first detection element 402 second detection element 403 third detection element

Claims (13)

1. It has at least a first detection element that detects glucose in a biological fluid and a second detection element that detects a biological substance other than glucose, and the second detection element includes a semiconductor element. Sensor.
2. The sensor according to claim 1, wherein the biological substance is glycated hemoglobin or glycated albumin.
3. 3. The sensor according to claim 1, further comprising a third detection element, wherein the third detection element detects hemoglobin.
4. 4. The sensor according to claim 3, wherein the third detection element includes a semiconductor element.
5. The sensor according to any one of claims 1 to 4, wherein a detection part area of the second detection element is twice or more a detection part area of the third detection element.
6. The sensor according to any one of claims 1 to 5, wherein the semiconductor element contains carbon nanotubes.
7. The semiconductor element includes at least a substrate, a first electrode, a second electrode, and a semiconductor layer, and the first electrode and the second electrode each include a metal type of 80 wt% or more, a double-walled carbon nanotube, The sensor according to any one of claims 1 to 6, comprising at least one selected from multi-walled carbon nanotubes.
8. The sensor according to any one of claims 1 to 7, wherein the semiconductor layer contains carbon nanotubes.
9. The sensor according to any one of claims 6 to 8, wherein a conjugated polymer is attached to at least a part of the surface of the carbon nanotube.
10. 10. The sensor according to claim 7, wherein another biological substance that selectively interacts with the biological substance is immobilized on the semiconductor layer.
11. The sensor according to any one of claims 1 to 10, wherein the sensor has a biological fluid inlet, and a path connecting the inlet to the first detection element and the second detection element. Sensor.
12. The sensor according to any one of claims 1 to 11, wherein the biological fluid is blood.
13. The sensor according to any one of claims 1 to 12, wherein the path has a space for mixing the blood and a hemolytic agent between the blood inlet and the second detection element.

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