JP2005233808A - Manufacturing method for biosensor adjacent-bonded sheet - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a simple and convenient manufacturing method for a biosensor adjacent-bonded sheet suitable for mass production without requiring complicated positioning. <P>SOLUTION: In this method for obtaining the biosensor adjacent-bonded sheet including two or more of biosensors by bonding an electric insulating sheet-like substrate 1 and an electric insulating sheet-like cover 2 with a longitudinal direction as an X-axial direction and a short direction as a Y-axial direction in each thereof, via an adhesive layer 3, at least one set of electrodes 4 parallel or orthogonal to the X-axial direction is formed in a bonded face side of the sheet-like substrate, at least one sample conveying groove 24 pattern-formed with the adhesive layer is formed therein, two or more of at least ones out of the set of electrodes and the sample conveying grooves exist, the X-axial direction of the sheet-like substrate is conformed each other with the X-axial direction of the sheet-like cover, the Y-axial direction is positioned to make the electrodes orthogonal to the sample conveying grooves, and the sheet-like substrate is bonded to the sheet-like cover. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a biosensor connection sheet manufacturing method or a biosensor connection sheet.

Conventionally, as disposable sensors (Patent Document 1 and Patent Document 3), a three-dimensional structure is taken in order to ensure quantitativeness, and a capillary solution (Patent Document 5 and Patent Document 6) is used to obtain a sample solution. A mechanism for automatically introducing the sensor into the sensor is known (Patent Document 7). For example, as shown in FIG. 1, the sensor having such a configuration is assembled by stacking a spacer 3 and a cover 2 on an electrically insulating substrate 1. An electrode pattern 4 is formed on the substrate, and an air discharge port 7 necessary for air necessary for capillary action is opened on the cover. A sample introduction port 6 provided with an air discharge port 7 on one side and a sample transport path 8 are formed by the substrate 1, the spacer 3, and the cover 2 for introducing a predetermined amount of sample liquid 12 into the detection portion by capillary action. In addition, each of these components needs to be punched into a predetermined shape in advance, and positioning for accurate superposition of the components in the three-dimensional processing is also required. For this reason, there has been a problem that the three-dimensional machining process becomes more complicated as the number of components increases.
Furthermore, when these sensors require application of reagents such as molecular identification elements and mediators (Patent Documents 2 and 4) or formation of a film (Patent Document 8) to avoid the influence of interfering substances. There was a problem that the process became more complicated.
JP 47-500 A JP-A-48-37187 JP-A-52-142585 JP-A-54-50396 JP 56-79242 A JP-T 61-502419 Japanese Patent Laid-Open No. 1-291153 Japanese Patent Laid-Open No. 3-202864

  The conventional sensor described above requires many processes and materials for manufacturing, and has to take a complicated structure. As a result, a large capital investment was required for the production line, the product yield was not sufficient, and the cost was high. Naturally, the environmental load at the time of material procurement and manufacturing was also large. Furthermore, the coefficient of variation (CV), which is an index of variations in the manufactured sensor characteristics, is not sufficient due to complicated processes, particularly alignment during substrate lamination.

In order to solve the above-mentioned problems, the present inventors have conducted intensive research, and successively fed out and adhered the sheet-like substrate and the sheet-like cover sequentially so that the sample transport groove is orthogonal to the electrode and overlaps the reagent layer. As a result, it has been found that a biosensor connection sheet in which a plurality of biosensors are connected can be obtained simply and in large quantities without requiring complicated alignment, and the present invention has been completed.
That is, the present invention includes the following.

[1] The method for producing a biosensor connecting sheet according to the present invention includes an electrically insulating sheet-like substrate in which the longitudinal direction is the X-axis direction and the short direction is the Y-axis direction, and the longitudinal direction is the X-axis direction. By bonding an electrically insulating sheet-like cover whose short direction is the Y-axis direction via an adhesive layer,
A biosensor including an electrically insulating substrate, an electrode formed on the substrate, an adhesive layer, an electrically insulating cover coupled to the substrate through the adhesive layer, and a sample transport path; A method for obtaining a biosensor connecting sheet comprising two or more,
At least one set of positive and negative electrodes that are parallel or orthogonal to the X-axis direction is opposed to and formed on the adhesive surface side of the sheet-like substrate,
An adhesive layer is provided on the adhesive surface side of the sheet-like substrate or the sheet-like cover, and at least one sample transport groove patterned by the adhesive layer is formed in a direction perpendicular to the electrodes,
There are two or more of at least one of the set of electrodes or the sample transport groove,
The sheet-like substrate and the X-axis direction of the sheet-like substrate are aligned with the X-axis direction of the sheet-like cover, and the Y-axis direction is positioned so that the electrode and the sample transport groove intersect, It is characterized by adhering a sheet-like cover.
In the present invention, “one set of electrodes” includes not only a so-called two-pole method but also electrodes of a three-pole method or more.
[2] It is preferable that the sheet-like substrate and the sheet-like cover are continuously fed out in the X-axis direction and bonded.
[3] Two or more sets of the one set of electrodes may be formed in the Y-axis direction of the sheet-like substrate, and one sample transport groove may be formed in the X-axis direction of the sheet-like cover.
[4] The one set of electrodes is formed in one set in the X-axis direction of the sheet-like substrate,
Two or more sample transport grooves may be formed in the Y-axis direction of the sheet-like cover.
[5] Two or more sets of the one set of electrodes may be formed in the Y-axis direction of the sheet-like substrate, and two or more of the sample transport grooves may be formed in the X-axis direction of the sheet-like cover.
[6] Two or more sets of the one set of electrodes may be formed in the X-axis direction of the sheet-like substrate, and two or more of the sample transport grooves may be formed in the Y-axis direction of the sheet-like cover.
[7] Each electrode formed in a direction parallel to the X-axis direction or each sample transport groove formed in a direction parallel to the X-axis direction may be continuously formed in the X-axis direction.
[8] There is a reaction detection part further coated with a reagent layer on part or all of the surface of each set of electrodes,
The X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover are made to coincide, the electrode and the sample transport groove intersect, and the reagent layer and the sample transport groove overlap each other. The sheet-like substrate and the sheet-like cover can be bonded together.
[9] In the biosensor connection sheet manufacturing method of the present invention, the adhesive layer and the sample transport groove may be formed on the surface of the sheet-like substrate, and the substrate and the cover may be bonded. it can.
[10] In the method for manufacturing a biosensor connection sheet of the present invention, the adhesive layer and the sample transport groove may be formed on the surface of the sheet-like cover, and the substrate and the cover may be bonded. More preferred.
[11] On the same surface of the sheet-like substrate or the sheet-like cover, a sample introduction port and an air discharge port communicating with the sample transport groove are provided, and the sample introduction port and the air discharge port sandwich the electrode. You may adhere | attach so that it may oppose.
[12] On the same surface of the sheet-like substrate or the sheet-like cover, at least one sample introduction port and at least two air discharge ports communicating with the sample conveyance path are provided, the sample introduction port and the air You may adhere | attach so that a discharge port may oppose on both sides of an electrode.
[13] A sample introduction port and an air discharge port communicating with the sample transport path may be included in a cross section of the adhesive layer sandwiched between the substrate and the cover.
[14] The cross section of the adhesive layer sandwiched between the substrate and the cover may include at least one sample inlet and at least two air outlets communicating with the sample transport path.
[15] The biosensor connecting sheet according to the present invention comprises an electrically insulating substrate, an electrode formed on the substrate, an adhesive layer, and an electrically insulating material bonded to the substrate through the adhesive layer. A biosensor connecting sheet including two or more biosensors including a cover and a sample conveyance path,
An electrically insulating sheet-like substrate whose longitudinal direction is the X-axis direction and the short direction is the Y-axis direction, and an electrically insulating sheet-like cover whose longitudinal direction is the X-axis direction and whose transverse direction is the Y-axis direction And an electrode formed on the substrate, an adhesive layer, and a sample transport path,
At least one set of positive and negative electrodes parallel or orthogonal to the X-axis direction is opposed to the surface on the adhesive surface side of the sheet-like substrate,
An adhesive layer is formed on the adhesive surface side of the sheet-like substrate or the adhesive surface side of the sheet-like cover, and the sample transport groove patterned by the adhesive layer is oriented in a direction perpendicular to the electrodes. At least one formed,
There are two or more of at least one of the set of electrodes or the sample transport groove,
The X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover coincide with each other, and the electrode and the sample transport groove intersect each other.
[16] In the biosensor connecting sheet, each electrode formed in the X-axis direction and / or each sample transport groove formed in the X-axis direction may be provided continuously in the X-axis direction. preferable.
[17] A reaction detection part further coated with a reagent layer is present in the vicinity of each set of electrodes or part or all of the electrode surface,
Even if the X-axis direction of the sheet-like substrate coincides with the X-axis direction of the sheet-like cover, the electrode and the sample transport groove intersect, and the reagent layer and the sample transport groove overlap each other. Good.

  According to the biosensor connecting sheet manufacturing method of the present invention, the X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover are matched, and the electrode and the sample transport groove intersect each other. By simply positioning the Y-axis direction and adhering the sheet-like substrate and the sheet-like cover, a biosensor connection sheet in which a plurality of biosensors are connected can be easily obtained without requiring complicated alignment. be able to. This makes it possible to manufacture a biosensor that is excellent in productivity and economy and has a low environmental load.

  Hereinafter, the biosensor manufacturing method according to the present invention will be described with respect to raw materials such as a substrate, a cover, an adhesive layer, an electrode, a reagent layer, and a manufacturing method.

Substrate, cover, adhesive layer, sample transport groove, electrode, reagent layer
Substrate, cover The electrically insulating substrate that can be used in the present invention has characteristics required for use as a biosensor, such as chemical resistance, heat resistance, bending resistance, and dimensional stability. However, it only needs to be electrically insulating. The shape is not limited, but examples thereof include a plate shape and a sheet shape. In the case of the sheet shape, those that can be wound into a roll shape are preferable.

Although the thickness of the said board | substrate is not specifically limited, For example, Preferably it is 10-2000 micrometers, More preferably, it is 100-500 micrometers.
The width of the substrate varies depending on the allowable width of the biosensor to be manufactured and the apparatus used for bonding, and is not limited, but it may be generally in the range of about 0.3 cm to 2 cm.

As such a substrate, for example, plastic, biodegradable material, or paper can be preferably used.
Examples of the plastic include polyethylene terephthalate, polyethylene naphthalate, polyether nitrile, polycarbonate, polyamideimide, and phenol resin. When used in a sheet form, polyethylene terephthalate or the like can be preferably used.
The biodegradable material is preferably polylactic acid.

As the electrically insulating cover, the same material as the substrate can be used.
Although the thickness of the said cover is not specifically limited, For example, Preferably it is 10-2000 micrometers, More preferably, it is 100-500 micrometers.

Perforations and cuts for separating the biosensor may be provided on both or one of the substrate and the cover. Further, it is preferable that the perforations or cuts are formed on a substrate or sheet on which two sets (or two) or more of the one set of electrodes or sample transport grooves are formed. The position of the perforation or cut may be a position that separates a set of electrodes or sample transport grooves formed in two sets (or two) or more.
Of the substrate and the cover, it is preferable that a perforation or a cut is provided on one of the substrate and the cover. If it is provided on one side, it becomes a mark when the biosensor connecting sheet is separated into each biosensor, and the positioning in the manufacturing process is reduced, so that a simpler manufacturing method can be obtained.

On the other hand, when perforations or cuts are provided in both the substrate and the sheet, there is an advantage that alignment in the manufacturing process can be easily performed.
There are no particular limitations on the shape and dimensions of the perforations and cuts, and it is only necessary to maintain strength that does not separate during the manufacturing process.

Electrodes Examples of electrode materials that can be used in the present invention include metals such as carbon, platinum, gold, silver / silver chloride, silver, copper, palladium, iridium, lead, nickel, titanium, tin oxide, and platinum black. . These are excellent in conductivity.
Although the thickness of the electrode is not limited as long as it is smaller than the thickness of the adhesive layer, it is usually about 200 to 2000 angstrom, preferably about 500 to 1000 angstrom. When the carbon electrode is formed on the substrate, the thickness of the electrode is not limited as long as it is smaller than the thickness of the adhesive layer, but it is usually about 1 to 100 μm, preferably about 3 to 20 μm.
In such a range, the edge of the electrode formed on the substrate does not have a saw-like shape, and the electrode is highly accurate. In addition, electrode peeling and disconnection can be prevented.

  Such an electrode can be easily patterned on a substrate by vapor deposition, sputtering, plating, CVD, coating and drying, and the like.

  As the carbon, carbon nanotubes, carbon microcoils, carbon nanohorns, fullerenes, dendrimers, and derivatives thereof can also be used. Moreover, it is preferable to use these carbon powders. These are suitable for immobilization of molecular identification elements and electrode materials because of their characteristics (structure, conductivity, etc.).

  The carbon powder is slightly inferior in conductivity compared to platinum and gold, but it can be easily formed by a screen printing method or the like, similar to silver powder, by applying a paste or the like to a substrate. . Furthermore, fine particles such as platinum and gold can be pasted and processed by printing.

In the case of a protein sensor, nickel is preferably used as the electrode material. Nickel oxidizes the amino group of a protein under a predetermined condition to become a protein sensor. FIA (flow injection analysis) is also possible.
The electrodes form a pair of electrodes in which the + electrode and the − electrode face each other.
Such an electrode may be composed of two electrodes consisting of + and −, or may be two or more.
The electrodes may form an array. In the present specification, “array” means being arranged in an array.

Adhesive Layer, Sample Conveying Groove As the adhesive forming the adhesive layer that can be used in the present invention, an acrylic resin-based one can be used.
Among these, a thermosetting resin or a photocurable resin can be preferably used.
Although the thickness of the said adhesive bond layer is not specifically limited, For example, Preferably it is 5-500 micrometers, More preferably, it should just exist in the range of about 10-100 micrometers.

  The depth of the flow path conveying groove corresponds to the thickness of the adhesive layer, and is preferably in the range of about 5 to 500 μm, more preferably about 10 to 100 μm. The width of the groove is preferably 50 to 5000 μm, more preferably 300 to 2000 μm, and more preferably 500 to 1000 μm.

  Such a channel conveying groove or channel channel is patterned by an adhesive layer. The adhesive layer is preferably formed by screen printing so as to form a desired flow path conveyance path.

  It is also possible to include a reagent in the adhesive layer and simultaneously form the adhesive layer and, if necessary, the reagent layer by screen printing.

Reagent layer When a reagent layer is used in the present invention, the reagent layer may be an enzyme, an antibody, a nucleic acid, a primer, a peptide nucleic acid, a nucleic acid probe, a microorganism, an organelle, a receptor, a cell tissue, a molecular identifier such as crown ether, a mediator, Intercalating agents, coenzymes, antibody labeling substances, substrates, inorganic salts, surfactants, lipids and the like can be included. These can be used singly or in combination.
When the sample solution is blood or the like, heparin, prolyxin-S, ethylenediaminetetraacetic acid, a metal salt of citric acid, or the like may be included as an anticoagulant.

  Examples of the enzyme include enzymes such as oxidase or dehydrogenase, such as glucose oxidase, fructosylamine oxidase, lactate oxidase, urate oxidase, cholesterol oxidase, alcohol oxidase, glutamate oxidase, pyruvate oxidase, glucose dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, cholesterol Examples include esterase, protease, and DNA polymerase. These enzymes can be used alone or in combination.

  Further, the reagent layer may be contained in combination with a mediator instead of the enzyme alone. This mediator is selected from potassium ferricyanide, ferrocene, and benzoquinone. The reagent layer may contain a combination of inorganic salts such as sodium chloride and potassium chloride and quinhydrone.

  The reagent layer may contain a combination of primer, DNA polymerase, and deoxyribonucleotide triphosphate. Further, the reagent layer may contain a combination of inorganic salts such as sodium chloride and potassium chloride and quinhydrone in primer, DNA polymerase, deoxyribonucleotide triphosphate.

  When using a biosensor as a DNA chip, a nucleic acid probe can be immobilized as a reagent layer. In this case, it is preferable to arrange the electrodes in an array.

  In the present invention, the surface of the reagent layer can be coated with a compound that facilitates movement of the sample liquid such as a surfactant and lipid. If the surface of the reagent layer is coated with a surfactant or the like, deterioration due to air oxidation can be further suppressed. When the sample solution is blood or the like, heparin, prolixin-S, ethylenediaminetetraacetic acid, a metal salt of citric acid, or the like may be coated as an anticoagulant.

Although the thickness of a reagent layer is not specifically limited, For example, Preferably it is 0.05-300 micrometers, More preferably, it is 1-100 micrometers.
The reagent layer is formed on a part or all of the surface of each set of electrodes, and constitutes a reaction detection unit together with the electrodes.

Such a reagent layer can be formed by a dispenser method in which the reagent layer is dropped by a dispenser or the like and dried, a screen printing method in which the viscosity is adjusted, and the like, and among these, the dispenser method is preferable. The reagent layer can be immobilized on the electrode surface or substrate surface by an adsorption method involving drying or a covalent bonding method.
The reagent layer is not limited to one place, and two or more places can be provided. In that case, two or more kinds of different reagent layers may be provided.

When there is only one reagent layer, the number of substances to be detected is one. However, when two kinds of substances are detected at the same time, for example, as shown in FIG. It can also be formed on a substrate (for example, JP-A-1-291153).
When two or more reagent layers are provided, as shown in FIG. 10, a convex partition is provided between them in order to prevent mixing of the dissolved reagent layers in the middle of the reagent layer. You can also. The convex partition portion can be formed by a screen printing method using carbon, a resist, a water-absorbing material, or the like.

In this case, the initial thickness of the convex portion should be thinner than the adhesive layer, and when bent, the upper and left and right sides of the convex portion must be open. This is to facilitate the passage of the sample.
When the convex partition is made of a water-absorbing material, it can have a function of preventing mutual dissolution reagents from mixing due to swelling after passing through the sample. Further, when the number of electrodes is not four (FIG. 20) but three, it is not shown, but for example, the middle one can be used as a common counter electrode.

The present invention provides a method for producing a biosensor that monitors an electrical change in an electrode by bringing a sample fed from a sample transport path into contact with the electrode or a reagent layer on the electrode.
When a nickel electrode is used as the electrode, it can be used as a protein sensor (for example, US Pat. No. 5,653,864) that detects protein even without a reagent layer. When a platinum electrode is used as the electrode, this sensor can be used as an oxygen sensor by combining a conductivity sensor, a hydrogen peroxide sensor, an oxygen permeable membrane, and an electrolyte.

On the other hand, when a reagent layer is used, blood glucose sensor, urine sugar sensor, glycated hemoglobin sensor (for example, JP-A-2001-204494), lactate sensor, uric acid sensor, cholesterol sensor, alcohol sensor, glutamic acid sensor using an enzyme and a mediator , Pyruvate sensor, phosphate sensor, pyrophosphate sensor, silver / silver chloride electrode and quinhydrone, pH sensor using inorganic salt (for example, Japanese Patent Laid-Open No. 9-222414), pH sensor and primer, DNA polymerase, etc. Various biosensors such as single nucleotide polymorphism sensors and DNA chips using immobilized nucleic acid probes can be manufactured, and can be applied as sensors for detecting various chemical and physical states.

Manufacturing method of biosensor connecting sheet The manufacturing method of the biosensor connecting sheet according to the present invention includes an electrically insulating sheet-like substrate in which the hand direction is the X-axis direction and the short direction is the Y-axis direction, and the longitudinal direction is X. By adhering the electrically insulating sheet-like cover whose axial direction is the Y-axis direction via the adhesive layer,
A biosensor including an electrically insulating substrate, an electrode formed on the substrate, an adhesive layer, an electrically insulating cover coupled to the substrate through the adhesive layer, and a sample transport path; A method for obtaining a biosensor connecting sheet comprising two or more,
On the adhesive surface side of the sheet-like substrate, at least one set of positive and negative electrodes parallel or orthogonal to the X-axis direction is opposed, and is formed.
An adhesive layer is provided on the adhesive surface side of the sheet-like substrate or the sheet-like cover, and at least one sample transport groove patterned by the adhesive layer is formed in a direction perpendicular to the electrodes,
There are two or more of at least one of the set of electrodes or the sample transport groove,
The sheet-like substrate and the X-axis direction of the sheet-like substrate are aligned with the X-axis direction of the sheet-like cover, and the Y-axis direction is positioned so that the electrode and the sample transport groove intersect, It is characterized by adhering a sheet-like cover.

  In this specification, the longitudinal direction of the sheet-like substrate and the sheet-like cover is referred to as the X-axis direction, and the short direction of the sheet-like substrate and the sheet-like cover that is perpendicular to the X-axis direction is referred to as the Y-axis direction.

According to such a method, the X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover coincide with each other, and the position in the Y-axis direction is adjusted so that the electrode and the sample transport groove intersect. And it is possible to manufacture a biosensor connecting sheet including a plurality of biosensors simply by adhering the sheet substrate and the sheet cover. In particular, when either the electrode or the sample transport groove is continuously formed in the X-axis direction, alignment in the X-axis direction is not necessary.
The sample transport groove becomes a sample transport path by bonding the sheet-shaped substrate and the sheet-shaped cover together.

Usually, in biosensor manufacturing, for example,
1) Electrode formation on the substrate (normal formation conditions)
2) Bonding of a sheet-like cover on which an adhesive layer and a reagent layer are formed and an electrode substrate 3) Crimping 4) Cutting out a sensor 5) Drying 6) Packaging steps.

  In the production method of the present invention, the sheet-like substrate and the sheet-like cover may be bonded to each other. However, when the steps 1) and 2) are performed, ii) A mode in which the sheet-like substrate and the sheet-like cover are respectively wound in a roll shape, and members each stretched in a planar shape from two rolls are continuously bonded together. Among these, the mode (ii) is preferable. That is, it is preferable that the sheet-like substrate and the sheet-like cover are continuously fed out in the X-axis direction and bonded.

When forming in the form of a roll of (ii), an apparatus capable of continuously bonding the sheet-like substrate and the sheet-like cover fed out from the roll continuously at a constant speed by pressure bonding can be used.
The feeding speed of the sheet-like substrate and the sheet-like cover is preferably 1 to 50 cm / s, more preferably about 5 to 20 cm / s.
By setting it as such a speed range, a sample conveyance groove | channel is not crushed and a board | substrate and a cover can be adhere | attached with appropriate adhesiveness.
Bonding is preferably performed by pressure bonding with an adhesive layer. The pressure for crimping is not particularly limited as long as the biosensor is not damaged, and the substrate and the cover are in close contact with each other. For example, the pressure may be in the range of about 0.5 to ¹⁰ kg / cm ² .
The temperature at the time of pressurization depends on the substrate used and the material of the cover, but is preferably 5 to 70 ° C, more preferably 30 to 50 ° C (with enzyme), or preferably 50 to 200 ° C, more preferably 100. It is the range of -150 degreeC (electrode pattern formation to the board | substrate without an enzyme).

  In the case of (ii), even if (a) the roll to be used already has electrodes, an adhesive layer, and a channel conveying groove, (b) the substrate or cover is made flat from the roll. You may form an electrode, an adhesive bond layer, and a flow-path conveyance groove | channel in the extended state.

  In the case of (a), it is preferable that an electrode pattern parallel to the Y-axis direction of the roll is formed and wound into a roll shape. Moreover, it is preferable to coat a release paper, for example, on the back surface of the substrate on the electrode pattern. Thereby, the adhesion between the electrode pattern and the sheet substrate can be prevented. Moreover, when forming the electrode pattern parallel to the X-axis direction of a roll, it is preferable to form a metal thin film on a tape-shaped plastic substrate. Thereby, distortion of an electrode can be prevented.

In the case of (b), an electrode pattern can be formed on a substrate extended from a roll onto a plane by a method such as vapor deposition or screen printing.
Among the above, it is preferable that (b) the electrode and the adhesive layer be formed into a pattern when the flat surface is extended from the roll, and then bonded together.

  About the method of bonding, for example, regarding the sheet-like cover, an adhesive layer (with a reagent transport path formed as necessary) and a reagent layer are formed inside the roll-like sheet-like cover, and the above-mentioned The method of bonding with the board | substrate on a sheet | seat formed at the process of 1) is mentioned.

  In the present invention, for example, each electrode formed in a direction parallel to the X-axis direction or each sample transport groove formed in a direction parallel to the X-axis direction is continuously formed in the X-axis direction. In this case, positioning in the X-axis direction can be eliminated, and extremely efficient continuous production of sensor chips is possible.

  On the other hand, if not in the above-described manner, it goes without saying that in the present invention, the sheet-like cover and the sheet-like substrate may be positioned as necessary. In this case, for example, it is preferable that positioning holes or positioning marks are provided on one side of the sheet or on both sides of the sheet in the X-axis direction of the sheet-like cover and the sheet-like substrate. Also in this case, continuous mass production of sensor chips becomes possible.

More specific embodiments of the electrode and the sample transport groove include the following embodiments (1) to (4).
(1) A mode in which two or more sets of the one set of electrodes are formed in the Y-axis direction of the sheet-like substrate, and one sample transport groove is formed in the X-axis direction of the sheet-like cover. (For example, FIG. 2, FIG. 10, etc.)
(2) A mode in which one set of the one set of electrodes is formed in the X-axis direction of the sheet-like substrate, and two or more sample transport grooves are formed in the Y-axis direction of the sheet-like cover. (For example, FIG. 4, FIG. 6, FIG. 8, FIG. 9 etc.)
(3) A mode in which two or more sets of the one set of electrodes are formed in the Y-axis direction of the sheet-like substrate, and two or more of the sample transport grooves are formed in the X-axis direction of the sheet-like cover. (For example, FIG. 11 etc.)
(4) A mode in which two or more sets of the one set of electrodes are formed in the X-axis direction of the sheet-like substrate, and two or more of the sample transport grooves are formed in the Y-axis direction of the sheet-like cover. (For example, FIG. 13, FIG. 15, FIG. 17, etc.)

  Which aspect of the manufacturing method is used depends on the type of biosensor desired and is not limited.

Each electrode formed in a direction parallel to the X-axis direction and each sample transport groove formed in a direction parallel to the X-axis direction are preferably formed continuously in the X-axis direction.
For example, in the aspect (1), it is preferable that the sample transport groove formed in the X-axis direction of the sheet-like cover is continuously provided in the X-axis direction.
In the aspect (2), it is preferable that one set of electrodes formed in the X-axis direction of the sheet-like substrate is continuously provided in the X-axis direction.
In the aspect (3), it is preferable that two or more sample transport grooves formed in the X-axis direction of the sheet-like cover are continuously provided in the X-axis direction.
In the aspect (4), it is preferable that two or more pairs of electrodes formed in the X-axis direction of the sheet-like substrate are continuously provided in the X-axis direction.
Thus, when the electrode or the sample transport groove is continuously provided in the X-axis direction, there is no need to align in the X-axis direction.

In addition, a reaction detection unit to which a reagent layer is further applied may be present in the vicinity of each set of electrodes or part or all of the electrode surface. In this case, the X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover are made to coincide, the electrode and the sample transport groove intersect, and the reagent layer and the sample transport groove overlap. In addition, the sheet-like substrate and the sheet-like cover can be bonded by positioning in the Y-axis direction.
Therefore, also in this case, the X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover are made to coincide, the electrode and the sample transport groove intersect, and the reagent layer and the sample transport groove The biosensor connecting sheet including a plurality of biosensors can be easily manufactured simply by positioning so as to overlap each other.
Moreover, in this invention, it can obtain as a sheet | seat where the some biosensor is connected.

In this invention, the said adhesive layer and the said sample conveyance groove | channel may be formed in the said sheet-like board | substrate surface, and the said sheet-like board | substrate and the said sheet-like cover may be adhere | attached, The sample transport groove may be formed on the surface of the sheet-like cover, and the sheet-like substrate and the sheet-like cover may be bonded. Of these, the latter method is preferred. By forming the electrodes on the substrate and forming the sample transport groove on the cover, the yield can be improved and the working efficiency can be improved.
The reagent layer may be mixed with an adhesive used as a spacer material.

  In the present invention, a sample introduction port and an air discharge port communicating with the sample transport path can be provided on the same surface of the sheet substrate or the sheet cover. Alternatively, on the same surface of the sheet-like substrate or the sheet-like cover, at least one sample introduction port and at least two air discharge ports communicating with the sample transport path may be provided. Also in this case, it is only necessary to form the sample inlet and the air outlet on the substrate or cover in advance so that the sample inlet and the air outlet are opposed to each other with the sample transfer groove therebetween, and the burden of alignment is greatly reduced. To do.

Furthermore, in the present invention, a sample introduction port and an air discharge port communicating with the sample transport path are included in the cross section of the adhesive layer sandwiched between the sheet-like substrate and the sheet-like cover. When the biosensor is separated, the sample inlet and the air outlet may be exposed. Further, at least one sample introduction port and at least two air discharge ports communicating with the sample conveyance path are included in the cross section of the adhesive layer sandwiched between the sheet substrate and the sheet cover. When the biosensor is used, the sample introduction port and the air discharge port may be exposed when each biosensor is separated.
In this case, in actual use, the sample introduction port and the air discharge port are exposed by cutting each biosensor from the biosensor connection sheet, so that it is easy to maintain the sealed state of the sample transport path. is there. Moreover, since it is not necessary to align the sample introduction port, the air discharge port, and the electrode, the production efficiency is remarkably high.

  As long as such a sample introduction port is a position where a sample can be injected into the sample conveyance path, it may be one end or an intermediate point of the sample conveyance path.

  In the above invention, a surfactant or lipid may be applied to the periphery of the sample introduction port and the surface of the sample transport groove. By applying a surfactant or lipid, the sample can be moved smoothly. Moreover, the tip part of the sample introduction port can have a structure having a curved part.

  A part of the adhesive layer may be a non-adhesive region, and a part of the non-adhesive region may form an exposed portion for terminal connection. The non-adhesive region forming the terminal connection exposed portion may be a non-adhesive layer or a removable adhesive layer.

  In the present invention, a biosensor connection sheet including a plurality of biosensors can be obtained. The substrate and the cover at the boundary portion of each biosensor are provided with perforations or cuts for separating each biosensor in advance. And the cover are preferably aligned.

Biosensor connection sheet The biosensor connection sheet according to the present invention includes an electrically insulating substrate, an electrode formed on the substrate, an adhesive layer, and an electrically insulating material that is bonded to the substrate through the adhesive layer. A biosensor connecting sheet including two or more biosensors including a cover and a sample transport path,
An electrically insulating sheet-like substrate whose longitudinal direction is the X-axis direction and the short direction is the Y-axis direction, and an electrically insulating sheet-like cover whose longitudinal direction is the X-axis direction and whose transverse direction is the Y-axis direction And an electrode formed on the substrate, an adhesive layer, and a sample transport path,
At least one set of positive and negative electrodes parallel to or orthogonal to the X-axis direction is opposed to the surface on the adhesive surface side of the sheet-like substrate,
An adhesive layer is formed on the adhesive surface side of the sheet-like substrate or the adhesive surface side of the sheet-like cover, and the sample transport groove patterned by the adhesive layer is oriented in a direction perpendicular to the electrodes. At least one formed,
There are two or more of at least one of the set of electrodes or the sample transport groove,
The X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover coincide with each other, and the electrode and the sample transport groove intersect each other.

In such a biosensor connecting sheet, each electrode formed in the X-axis direction or each sample transport groove formed in the X-axis direction is preferably provided continuously in the X-axis direction.
Further, there is a reaction detection unit coated with a reagent layer in the vicinity of each set of electrodes or in part or all of the electrode surface, and the X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover May coincide, the electrode and the sample transport groove intersect, and the reagent layer and the sample transport groove may overlap.
Moreover, various aspects and constituent materials described in the method for producing the biosensor connecting sheet can be employed.

For example, in the biosensor connecting sheet of the present invention, the more specific embodiments of the electrode and the sample transport groove include the following embodiments (1) to (4) as described above.
(1) A mode in which two or more sets of the one set of electrodes are formed in the Y-axis direction of the sheet-like substrate, and one sample transport groove is formed in the X-axis direction of the sheet-like cover. (For example, FIG. 2, FIG. 10, etc.)
(2) A mode in which one set of the one set of electrodes is formed in the X-axis direction of the sheet-like substrate, and two or more sample transport grooves are formed in the Y-axis direction of the sheet-like cover. (For example, FIG. 4, FIG. 6, FIG. 8, FIG. 9 etc.)
(3) A mode in which two or more sets of the one set of electrodes are formed in the Y-axis direction of the sheet-like substrate, and two or more of the sample transport grooves are formed in the X-axis direction of the sheet-like cover. (For example, FIG. 11 etc.)
(4) A mode in which two or more sets of the one set of electrodes are formed in the X-axis direction of the sheet-like substrate, and two or more of the sample transport grooves are formed in the Y-axis direction of the sheet-like cover. (For example, FIG. 13, FIG. 15, FIG. 17, etc.)

  Furthermore, in the present invention, a sample introduction port and an air discharge port communicating with the sample transport path are included in the cross section of the adhesive layer sandwiched between the sheet-like substrate and the sheet-like cover. When the biosensor is separated, the sample inlet and the air outlet may be exposed. Further, at least one sample introduction port and at least two air discharge ports communicating with the sample conveyance path are included in the cross section of the adhesive layer sandwiched between the sheet substrate and the sheet cover. When the biosensor is used, the sample introduction port and the air discharge port may be exposed when each biosensor is separated.

The electrodes may form an array. In the biosensor forming the array in this manner, at least one sample introduction port exists on the same surface of the substrate or cover, and at least one set of electrodes is provided at the tip of the sample transport path communicating with the sample introduction port. It is preferable that they intersect.
The at least one sample introduction port may communicate with at least two sample conveyance paths branched from the sample introduction port, and may cross at least one pair of electrodes at the tip of the sample conveyance path.
In the present specification, “array” means being arranged in an array.

  When at least two sample liquid transport paths branch from one sample introduction port, the sample transport path may be coated with a surfactant so that the sample liquid can reach all the reagent layers in the array, or the sample When the liquid is blood or the like, heparin, prolyxin-S, ethylenediaminetetraacetic acid, a metal salt of citric acid or the like may be coated as an anticoagulant.

  Such biosensor connecting sheets can be used as individual biosensors by separating them. For example, the following usage method can be adopted by changing the type of the reagent layer.

  For example, in an enzyme sensor, the type of enzyme serving as a molecular identification element is changed depending on the subject to be measured. For example, glucose oxidase or glucose dehydrogenase when the measurement target is blood glucose (glucose), urine sugar, a mixture of fructosylamine oxidase and protease when the measurement target is glycated hemoglobin, lactate oxidase when the measurement target is lactic acid, measurement Mixture of cholesterol esterase and cholesterol oxidase when the target is total cholesterol, urate oxidase when the measurement target is uric acid, alcohol oxidase when the measurement target is ethanol, glutamate oxidase when the measurement target is glutamic acid, and pyruvine In the case of acid, pyruvate oxidase or the like is used.

  In the enzyme sensor, an electron carrier (mediator) is used together with the enzyme. As the mediator, potassium ferricyanide, ferrocene, ferrocene derivatives, nicotinamide derivatives, flavin derivatives, benzoquinone, quinone derivatives and the like are used.

  In the case of a pH sensor, an inorganic salt such as sodium chloride or potassium chloride and a quinhydrone reagent layer are used on a substrate provided with a silver / silver chloride electrode and another electrode. In this case, the change in the interelectrode potential is measured.

  In the case of single nucleotide polymorphism (SNPs) sensors (A. Ahmadian et al., Biotechniques, 32, 748, 2002), a mixture of primer, DNA polymerase, deoxyribonucleotide triphosphate, etc. is used as a reagent on the above pH sensor. The change in pH when the analyte DNA in the sample and the primer are complementary is measured.

  An immunosensor uses an antigen-antibody reaction, and for example, when measuring human serum albumin, anti-albumin is used as a molecular identification element. In an immunosensor, an interelectrode potential that varies with the formation of an antigen-antibody complex is measured.

  In the microorganism sensor, microorganisms such as Pseudomonas fluorescence (measuring object: glucose) and Trichosporon brassicae (measuring object: ethanol) are used as the molecular identification element. Since these microorganisms breathe oxygen (aerobic) or produce metabolites in an oxygen-free environment, they will capture oxygen respiration or metabolites electrically.

  In the organelle sensor, an organelle is used as a molecular identification element. For example, NADH can be measured by using mitochondrial electron transfer particles. As this principle, NADH is oxidized by mitochondrial electron transfer particles, and oxygen is consumed at this time. Therefore, NADH and NADPH can be measured using this oxygen as an index.

  In the receptor sensor, a receptor such as a cell membrane is used as a molecular identification element. Specimens include hormones and neurotransmission. As a measurement principle, a change in acceptance is converted into a potential and measured through an electrode.

  Tissue sensors use animal and plant tissues as molecular identification elements. As animal and plant tissues, for example, frog skin, animal liver slices, cucumbers, banana peels and the like can be used. As a measurement principle, for example, in a sodium sensor using frog skin tissue, the frog skin tissue selectively permeates sodium ions, and the potential of the skin tissue changes at that time. Find the amount.

  Another example of the application of the biosensor described here is a DNA chip (Japanese Patent Laid-Open No. 5-199898). Many kinds of single-stranded nucleic acid probes having complementarity to many kinds of target genes to be detected are immobilized on an array of electrodes (US 4225410) as shown in FIGS. One type of nucleic acid probe is immobilized on one electrode. In order to confirm the presence or absence of a large number of target genes, a single-stranded denatured gene sample and a nucleic acid probe are hybridized, and then specifically bound to a double-stranded nucleic acid and electrochemically An active double-stranded recognizer is added to After washing, the substrate is folded in the presence of buffer solution, the array electrode is the working electrode, the upper electrode is the counter electrode, and voltage is applied to each electrode sequentially. The insertion agent is oxidized and an oxidation current flows. No current due to the intercalating agent flows at the electrode where the double strand is not formed. Since the type of the nucleic acid probe is known at the position of the electrode where the current is generated, the presence or absence of the target gene and the qualitativeness are possible. In addition, as said double strand recognition body, metallointercalators, such as intercalators (inserting agent), such as acridine orange, a tris (phenanthroline) cobalt complex, etc. can be used.

When measurement is performed using the biosensor as described above, the biosensor is attached to the apparatus, and the electrical value generated in the biosensor is measured.
This apparatus includes a measurement unit that measures an electrical value of the biosensor and a display unit that displays the measured value. As a measuring method of the measuring unit, potential step chronoamperometry, coulometry, voltammetry, or the like can be used. In addition, this apparatus can be provided with a memory for storing measurement values. In addition, when managing the measurement values remotely, wireless means such as Bluetooth may be installed.

  Further, in the biosensor connection sheet of the present invention, the biosensor is housed by separating the biosensor independently while connecting a part of the substrate to the biosensor as a coupling belt without separating all the connected biosensors. be able to. In this case, it is preferable that the substrate portion serving as the coupling belt has a notch that can be bent for storage.

  EXAMPLES Hereinafter, although this invention is demonstrated using an Example, this invention is not limited to a present Example at all.

FIG. 29 shows an example in which a biosensor connection sheet according to the present invention is manufactured by continuously laminating a sheet-like substrate and a sheet-like cover.
The substrate on the sheet is sent out from the roller around which the sheet-like substrate is wound, and then the electrode pattern is printed by a screen printer. Further, the electrode pattern is formed by heating the electrode pattern. Next, with respect to the sheet-like cover, an adhesive layer (with a reagent transport path formed as necessary) and a reagent layer are formed inside the roll-like sheet-like cover, and the sheet is formed as described above. Affixed to the substrate. Finally, individual biosensors are punched out from the obtained biosensor connection sheet.

FIG. 2a shows a plan view of the electrode substrate before the biosensor connection sheet is assembled. One sheet of electrically insulating, flat and long plate member, that is, a part of the substrate 1, a plurality of patterns 4 including a set of electrodes on the surface thereof are regularly connected in the Y-axis direction. They are arranged in a state. Here, “the surface is flat” means that there are no irregularities formed by artificially molding, cutting, bonding, etching, or the like. On both sides of the pattern 4 including the connected electrodes, there are shown portions of cuts 5 for cutting off after assembling the biosensor connecting sheet.
This electrode pattern 4 is formed in the Y-axis direction and is orthogonal to the reagent transport groove 25 formed continuously in the X-axis direction shown in FIG. 2b. In addition, a reagent layer 10 can be provided on the electrode pattern 4 as necessary.

  FIG. 2b also shows a plan view of the cover 2 before assembling the biosensor connecting sheet, and is a single sheet of electrically insulating, flat and long plate member, that is, a part of the cover 2, and a bonding that bears a spacer on the surface. An agent layer 3 is formed. Between the two upper and lower adhesive layers 3, that is, a sample transport groove 24, which is a region where no adhesive layer exists, is continuously formed in the X-axis direction. The sample transport groove 24 is a groove for forming a sample transport path 8 to be described later.

  Such a substrate shown in FIG. 2a and a cover shown in FIG. 2b are bonded with the same X-axis direction. When the reagent layer 10 is formed, either the reagent layer 10 or the sample transport groove 24 is formed so that the reagent layer 10 and the sample transport groove 24 overlap with each other, and they are aligned and bonded.

  FIG. 2c shows a plan view of one biosensor separated from the biosensor connection sheet assembled according to the present invention. The surface of the substrate 1 on which the electrode 4 is formed overlaps the surface of the cover 2 on which the adhesive layer 3 is formed, thereby forming a biosensor connection sheet in a state where a plurality of biosensors are connected. The biosensor separated along the boundary line defined by the interval of the pattern 4 including the electrodes on the substrate 1, that is, the cut line 5 is shown.

  One of the features of the biosensor manufacturing method of the present invention is an assembly process in which the two substrates and the cover of the biosensor are simply bonded together, and the sample transport groove 24 is continuously formed in the X-axis direction. Therefore, the positioning of the substrate and the cover in the X-axis direction is not required, and only the positioning in the Y-axis direction is necessary. In general, since a plurality of stacking steps are required, it is not possible to assemble a biosensor only by such simple positioning.

  FIG. 2d is a diagram of FIG. 2c, and shows a configuration diagram when only the cover member is removed. A sample transport path 8 is orthogonal to the upper part of the two electrode patterns 4 formed on the substrate 1, and the orthogonal part is a reaction detector 9. After separating each biosensor, a sample introduction port 6 is formed at the left end of the sample transport path 8 and an air discharge port is formed at the right end on the cut surface of the biosensor. If necessary, the reagent layer 10 as illustrated may be formed.

  FIG. 2e shows an example of biosensor usage. In this figure, the sample solution 12 is introduced from the sample introduction port 6 and is conveyed through the sample conveyance path 8 to fill the reaction detection unit 9 where the electrodes 4 are orthogonal. At this time, the air discharged from the air discharge port 7 by the transport of the sample liquid is discharged. The electrical signal detected by the reaction detector 9 is sent to the measuring device via the terminal 11.

  FIG. 3 shows a cross-sectional view of the biosensor shown in FIG.

By applying the method of Example 1, another biosensor connecting sheet shown in FIG. 4 can be produced.
4A and 2B, two electrodes are continuously provided in parallel on the substrate in the X-axis direction (FIG. 4a (1)), and an adhesive layer patterned thereon is parallel to the Y-axis direction. A manufacturing process of the biosensor connection sheet shown in FIG. 4B is shown by attaching a long cover (FIG. 4A (2)) in the X-axis direction provided with the flow path conveying groove 25 and perforation.
In this case, the terminal part 11 is covered with the terminal protective cover 14 which is a part of the cover 2 in the manufactured articulated biosensor (FIG. 4 b). Here, the terminal protective cover 14 is not provided with an adhesive layer as shown in FIG. 4c (1). When the biosensor is used, the terminal protection cover 14 is folded along the perforation 13 (FIG. 4c (1)), folded (FIG. 4c (2)), or cut (FIG. 4c (3)). To expose the terminals.

In the manufacturing process of the biosensor connecting sheet, since the electrodes are continuously provided in the X-axis direction, positioning of the substrate and the cover in the X-axis direction is not required, and only the Y-axis direction needs to be adjusted. That is, it is only necessary to adopt a continuous processing method in which the two substrates and the cover are bonded together while being long. For this purpose, it is preferable that the substrate and the cover are both continuous and not cut. However, the terminal portion 11 is hidden inside the biosensor by the cover 2 when trying to adopt the structure. In order to overcome this problem, the cover 2 is provided with perforations 13 as shown in FIG. Thereby, the perforation 13 can perform both the role of maintaining the cover 2 with a long structure and the role of exposing the terminal 11 covered with the terminal protective cover 14 when the biosensor is used.
FIG. 5 shows a cross-sectional view of the biosensor shown in FIGS. 4c (1)-(2).

  By applying the method of Example 1, another biosensor connecting sheet shown in FIG. 6 can be produced. 6 and 7 can be manufactured by substantially the same method as FIG. 4 differs from the case shown in FIG. 4 in that two through holes 17 are formed in a space 18 without a spacer in the adhesive layer 3 of the cover 2, and as shown in FIG. 6c, these through holes are formed. Reference numeral 17 denotes a sample inlet 6 and an air outlet 7 when a biosensor is formed. That is, the sample inlet 6 and the air outlet 7 can be formed other than between the biosensor substrates.

  Furthermore, in the biosensor shown in FIG. 6c, the terminal 11 is covered with the terminal protective cover 14 as in FIG. 4c, and the terminal 11 is displayed by folding the terminal protective cover 14 according to the two perforations 13 during use. It is a structure that appears in In FIG. 7a, a minute adhesive portion 19 is provided for maintaining the shape of the terminal protection cover 14 once folded.

By applying the method of Example 1, another biosensor connecting sheet shown in FIG. 8 can be produced. FIG. 8 can be manufactured by substantially the same method as FIG. 4, the biosensors are formed in the same direction (see FIG. 4 c), as can be seen from the repeated pattern of the adhesive layer 3 of the cover 2 in FIG. 4, and the two biosensors are opposite in FIG. 8 c. It is formed to become.
Further, in FIG. 8c, only one perforation 13 for the terminal protection cover 14 is provided. Thus, the number of perforations 1 for the terminal protection cover 14 is not particularly limited.

By applying the method of Example 1, another biosensor connection sheet shown in FIG. 9 can be produced.
In FIG. 9, the difference from FIG. 4 is that the cover 2 shown in FIG. 9a (2) is regularly formed on the removable adhesive layer 15, so that the electrode shown in FIG. 9a (1) is formed. The terminal 11 is exposed to the biosensor formed by bonding with the substrate 1 provided with 4. In the manufacturing method of FIG. 9, the cover 2 is regularly formed on the surface of the adhesive layer 15 with a long tape or the like, thereby making it unnecessary to align the biosensor in the X-axis direction as in FIG. it can.

By applying the method of Example 1, another biosensor connecting sheet shown in FIG. 10 can be produced.
10 is formed by cutting the biosensor shown in FIG. 2 into two sets of electrodes. With such a structure, the reagent layer is not limited to one place but can be installed in two or more places. In that case, two or more kinds of different reagent layers may be provided. Moreover, when two or more reagent layers are provided, the convex partition part 20 can also be provided between these. The convex partition portion can be formed by a screen printing method. The partition part of this convex part can be comprised from either carbon, a resist, or a water absorbing material.
In FIG. 10, the perforation is provided on both the substrate and the cover, but either one may be provided (not shown). In either case, alignment is easier.

By applying the method of Example 1, another biosensor connecting sheet shown in FIGS.
FIGS. 11 to 18 show a case where the biosensor forming patterns provided on the substrate and the cover form a plurality of rows in the biosensor manufacturing method shown in FIGS. Specifically, FIGS. 11 and 12 are shown in FIG. 2, FIGS. 13 and 14 are shown in FIG. 4, FIGS. 15 and 16 are shown in FIG. 8, and FIGS. Each of them.

Another example of the biosensor connection sheet of the present invention that can be manufactured in the same manner as in Example 1 is shown in FIGS.
19 to 26 require positioning in the Y-axis direction and positioning in the X-axis direction, but a biosensor connecting sheet including a large amount of biosensors can be manufactured at one time.
This method shows a process of manufacturing a biosensor in which electrodes form an array, and a configuration diagram, a cross-sectional view, and a usage example of the manufactured array biosensor.

  In FIG. 19a, a plurality of patterns including 10 sets of electrodes 21 connected in series on a single long substrate are connected in parallel in a state where a plurality of patterns are connected in the left-right direction. When the cover including the adhesive layer of the cover 2 (FIG. 19b) overlaps with this, the connected biosensor shown in the plan view of FIG. 20a is manufactured. Here, in order to detach the sensor manufactured in a connected manner, the detachable portion is provided with a sufficient space for preventing damage to the electrode wiring 22 and the terminal 11. FIG. 20b shows a plan view and a cross-sectional view of the array-type biosensor manufactured in this way. In this array type biosensor, the electrode wiring 22 and the terminal 11 are located inside the substrate 1, and there is a sample introduction port 6 at the center of the biosensor sandwiched between the substrate 1 and the cover 2. The conveyance path 8 is formed orthogonal to the 10 sets of electrodes 21, and the air exhaust port 7 is provided at the center of the lower end of the cover 2. Examples of use of this array type biosensor are shown in FIGS.

  21 to 23, a plurality of sample transport paths 8 connected to one sample introduction port 6 are branched, and in each sample transport path 8, 10 sets of electrodes 21 in series are formed so as to be orthogonal to each other. The manufacturing process (FIG. 21a, b) of the array-type biosensor provided with the air discharge port 7, the development view of the array-type biosensor to be manufactured (FIG. 22a), the plan view (FIG. 22b), and the cross-sectional view ( FIG. 23) is shown.

  Similarly, FIGS. 24 to 26 show a manufacturing process of an array-type biosensor of 20 rows and 10 columns including a sample introduction port 6, a sample transport path 8, a reaction detection unit 9 including an electrode 21, and an air discharge port 7. (FIGS. 24a and 24b), and a developed view (FIG. 25a), a plan view (FIG. 25b), and a cross-sectional view (FIG. 26) of the array-type biosensor to be manufactured.

  Each of the two types of array-type biosensors shown in FIGS. 21 to 26 is manufactured by laminating a substrate 1 and a cover 2 that are regularly formed on a long tape having a desorbent layer. The This method is the same as the method shown in FIGS.

  In any of the three types of array-type biosensors shown in FIGS. 19 to 26, the reagent layer 10 can be provided on each electrode 21. For example, DNA having a different base sequence can be used as a reagent layer on each set of electrodes. Can be used to detect a plurality of DNA sequences including single nucleotide polymorphisms (SNPs). Since the biosensor of this form can take a certain amount of measurement sample liquid into the sample transport path 8, it has reproducibility of measurement, compared to the type in which the reaction is detected by dropping the sample liquid on a flat substrate, It is characterized by being less susceptible to evaporation and drying during measurement of trace sample liquids.

  27 and 28 show an application example of the manufacturing method of the biosensor according to the present invention, which is manufactured by continuously joining the long substrate 1 and the cover 2 together. 27a (1) and (2) are plan views of two long plate members. In FIG. 27a (1), a pattern 4 including electrodes is provided in a state of being connected to a plate member constituting the substrate 1, In FIG. 27a (2), an adhesive layer 3 for defining individual biosensors is provided in a patterned state as a spacer. In particular, the plate member forming the substrate 1 shown in FIG. 27a (1) includes a cut (cut portion) 5 around the substrate 1 except for a portion of the substrate 1 portion defining each biosensor. In parallel, a long plate member provided with perforations 13 at equal intervals is provided on the upper part in a state where it is connected. This portion serves as a foldable belt for articulating the biosensor, as shown in FIG. 27b. FIG. 27c (1) shows a part of the connected biosensor in a plan view, and FIG. 27c (2) shows an example of using the biosensor in a connected state. As shown in FIG. 28a, the biosensor manufactured in this way can be stored in a connected state, and can be detached at the time of use or after use (FIG. 28b).

  By making such a form of biosensor, a cassette-type biosensor can be realized by, for example, regularly connecting the biosensor in a connected state or storing it in a roll shape.

FIG. 1 shows the structure of a conventional biosensor. FIG. 2 shows an example of manufacturing a biosensor connecting sheet according to the present invention. a is a part of the bonding surface side of the substrate, b is a part of the bonding surface side of the cover, c is a view of the biosensor separated from the manufactured biosensor connecting sheet, d is a plan view when only the cover is removed, e shows an example of using a biosensor. FIG. 3 shows an AA ′ sectional enlarged view (FIG. 3a), a BB ′ sectional enlarged view (FIG. 3b), and a CC ′ sectional enlarged view (FIG. 3c) of the biosensor of FIG. 2d. FIG. 4 shows an example of manufacturing a biosensor connecting sheet according to the present invention. a is a part of a plan view of the bonding surface side of the substrate and the cover, b is a part of a plan view of the manufactured biosensor connecting sheet, and c is a usage example (1) to (3) of the separated biosensor. FIG. 5 shows an AA ′ cross-sectional enlarged view (FIG. 5a), a BB ′ cross-sectional enlarged view (FIG. 5b), and a CC ′ cross-sectional enlarged view (FIG. 5c) of the biosensor of FIG. 4c. FIG. 6 shows an example of manufacturing a biosensor connecting sheet according to the present invention. a is a part of a plan view of the bonding surface side of the substrate and the cover, b is a part of a plan view of the manufactured biosensor connecting sheet, and c is an example of use of the separated biosensor. FIG. 7: shows the AA 'cross-sectional enlarged view (FIG. 7a), BB' cross-sectional enlarged view (FIG. 7b), and CC 'cross-sectional enlarged view (FIG. 7c) of the biosensor of FIG. 6c. FIG. 8 shows an example of manufacturing a biosensor connecting sheet according to the present invention. a is a part of the plan view of the bonding surface side of the substrate and the cover, b is a part of the plan view of the manufactured biosensor connection sheet, and c is a usage example (1) to (4) of the separated biosensor. FIG. 9 shows an example of manufacturing a biosensor connecting sheet according to the present invention. a is a part of a plan view of the bonding surface side of the substrate and the cover, b is a part of a plan view of the manufactured biosensor connecting sheet, and c is an example of use of the separated biosensor. FIG. 10 shows a biosensor in which two sets of electrodes are provided with different reagent layers so that two items can be measured. a is a part of the bonding surface side of the substrate, b is a part of the bonding surface side of the cover, c is a view of the biosensor separated from the manufactured biosensor connecting sheet, d is a plan view when only the cover is removed, e shows an enlarged cross-sectional view of the biosensor of FIG. FIG. 11a shows a part of a plan view on the bonding surface side of the substrate in which the electrode patterns are connected in two rows in the Y-axis direction, and b shows a part of the plan view on the bonding surface side of the cover. FIG. 12a shows a part of a plan view of the biosensor connecting sheet manufactured by bonding the two plate members shown in FIG. 11, and b shows an example of use of the separated biosensor. FIG. 13a shows a part of the plan view of the bonding surface of the substrate in which the electrode patterns are connected in 10 rows, and b shows a part of the plan view of the bonding surface of the cover. FIG. 14a shows a part of a plan view of a biosensor connecting sheet manufactured by bonding a substrate and a cover in FIG. 13, and b shows an example of use of a separated biosensor alone. FIG. 15a shows a part of the plan view of the adhesion surface of the substrate where the electrode patterns are connected in 10 rows, and b shows a part of the plan view of the adhesion surface of the cover. FIG. 16a shows a part of a plan view of a biosensor connecting sheet manufactured by bonding a substrate and a cover in FIG. 15, and b shows a usage example of a separated biosensor alone. FIG. 17a shows a part of the plan view of the bonding surface of the substrate in which the electrode patterns are connected in five rows, and b shows a part of the plan view of the bonding surface of the cover. FIG. 18a shows a part of a plan view of a biosensor connecting sheet manufactured by bonding the substrate and cover of FIG. 17, and b shows an example of use of the separated biosensor. FIG. 19a shows a part of the plan view of the bonding surface of the substrate in which the array type electrode patterns are connected in five rows, and b shows a part of the plan view of the bonding surface of the cover. 20a is a part of a plan view of an array type biosensor connecting sheet manufactured by bonding the substrate and cover of FIG. 19, b is a plan view of a separated array type biosensor, and its AA ′ cross-sectional enlarged view , BB ′ cross-sectional enlarged view, c, d show examples of use of the biosensor. FIG. 21a is a part of a plan view of an adhesive surface in a state where a substrate having an array type electrode pattern is regularly arranged on a desorption layer, and b is a desorption layer where a cover having an adhesive layer patterned as a spacer is used as a spacer. A part of the top view of the adhesion surface in the state regularly arranged on is shown. FIG. 22a is a plan view of the substrate and cover of FIG. 21, and b is a plan view of an array type biosensor connecting sheet manufactured by bonding them. FIG. 23 is an AA ′ cross-sectional enlarged view (FIG. 23a), a BB ′ cross-sectional enlarged view (FIG. 23b), and a CC ′ cross-sectional enlarged view (FIG. 23c) of the array type biosensor connecting sheet shown in FIG. 22b. ). FIG. 24a is a part of a plan view of an adhesive surface in a state where a substrate having another array type electrode pattern is regularly arranged on a desorption layer, and b is a cover on which an adhesive layer is patterned as a spacer. A part of top view of the adhesion surface of the state regularly arranged on the desorption layer is shown. 25a is a plan view of the substrate and cover of FIG. 24, and b is a plan view of the array type biosensor connecting sheet. FIG. 26 shows an AA ′ cross-sectional enlarged view (FIG. 26a), a BB ′ cross-sectional enlarged view (FIG. 26b), and a CC ′ cross-sectional enlarged view (FIG. 26c) of the array-type biosensor shown in FIG. 25b. . FIG. 27 shows a biosensor connecting sheet that is used with the substrate connected. a is a part of a plan view of the biosensor connection sheet on the bonding surface side of the substrate and a part of a plan view of the cover on the bonding surface side, and b is a plan view of the biosensor connection sheet manufactured by bonding the substrate and the cover. C shows an example of using a separated biosensor alone. FIG. 28a shows an example of storing the biosensor connection sheet, and b shows an example of use of the biosensor connection sheet. FIG. 29 shows an example of manufacturing a biosensor connecting sheet according to the present invention by continuously bonding a sheet-like substrate and a sheet-like cover.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Board | substrate 2 Cover 3 Adhesive layer 4 Pattern 5 including an electrode Cut part 6 Sample inlet 7 Air outlet 8 Sample conveyance path 9 Reaction detection part 10 Reagent layer 11 Terminal 12 Sample liquid 13 Perforation 14 Terminal protective cover 15 Desorbent Layer 16 Biosensor body 17 Through-hole 18 Space 19 without spacer 19 Detachable adhesive 20 Protrusion 21 Electrode 22 Wiring 23 Spacer plate member 24 Sample transport groove

Claims (17)

An electrically insulating sheet-like substrate whose longitudinal direction is the X-axis direction and the short direction is the Y-axis direction, and an electrically insulating sheet-like cover whose longitudinal direction is the X-axis direction and whose transverse direction is the Y-axis direction Are bonded through an adhesive layer,
A biosensor including an electrically insulating substrate, an electrode formed on the substrate, an adhesive layer, an electrically insulating cover coupled to the substrate through the adhesive layer, and a sample transport path; A method for obtaining a biosensor connecting sheet comprising two or more,
At least one set of positive and negative electrodes parallel to or orthogonal to the X-axis direction is opposed to the surface on the adhesive surface side of the sheet-like substrate,
An adhesive layer is formed on the surface of the sheet-like substrate or the cover of the sheet-like cover, and at least one sample transport groove patterned by the adhesive layer is formed in a direction perpendicular to the electrodes. And
There are two or more of at least one of the set of electrodes or the sample transport groove,
The sheet-like substrate and the X-axis direction of the sheet-like substrate are aligned with the X-axis direction of the sheet-like cover, and the Y-axis direction is positioned so that the electrode and the sample transport groove intersect, A method for producing a biosensor connecting sheet, comprising bonding a sheet-like cover. A method for producing a biosensor connecting sheet, wherein the sheet-like substrate and the sheet-like cover are continuously fed out in the X-axis direction and bonded together. Two or more sets of the one set of electrodes are formed in the Y-axis direction of the sheet-like substrate,
The method according to claim 1, wherein one sample transport groove is formed in the X-axis direction of the sheet-like cover. The one set of electrodes is formed in one set in the X-axis direction of the sheet-like substrate,
The method according to claim 1 or 2, wherein two or more of the sample transport grooves are formed in the Y-axis direction of the sheet cover. Two or more sets of the one set of electrodes are formed in the Y-axis direction of the sheet-like substrate,
The method according to claim 1 or 2, wherein two or more of the sample transport grooves are formed in the X-axis direction of the sheet cover. Two or more sets of the one set of electrodes are formed in the X-axis direction of the sheet-like substrate,
The method according to claim 1 or 2, wherein two or more of the sample transport grooves are formed in the Y-axis direction of the sheet cover. Each electrode formed in a direction parallel to the X-axis direction or each sample transport groove formed in a direction parallel to the X-axis direction is formed continuously in the X-axis direction. Item 7. The method according to any one of Items 1 to 6. There is a reaction detection part further coated with a reagent layer on a part or all of the surface of each set of electrodes,
The X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover are made to coincide, the electrode and the sample transport groove intersect, and the reagent layer and the sample transport groove overlap each other. The method according to claim 1, wherein the sheet-like substrate and the sheet-like cover are bonded together. The method according to claim 1, wherein the adhesive layer and the sample transport groove are formed on the surface of the sheet-like substrate to bond the substrate and the cover. The method according to claim 1, wherein the adhesive layer and the sample transport groove are formed on the surface of the sheet-like cover to bond the substrate and the cover. A sample introduction port and an air discharge port communicating with the sample transport groove are provided on the same surface of the sheet-like substrate or the sheet-like cover, and the sample introduction port and the air discharge port are opposed to each other with the electrode interposed therebetween. The method according to claim 1, wherein the method adheres to the surface. On the same surface of the sheet-like substrate or the sheet-like cover, at least one sample inlet and at least two air outlets communicating with the sample transport path are provided, the sample inlet and the air outlet The method in any one of Claims 1-10 which adhere | attach so that it may oppose on both sides of an electrode. The method according to claim 1, wherein a sample introduction port and an air discharge port communicating with the sample transport path are included in a cross section of the adhesive layer sandwiched between the substrate and the cover. The cross section of the adhesive layer sandwiched between the substrate and the cover includes at least one sample inlet and at least two air outlets communicating with the sample transport path. The method of crab. A biosensor including an electrically insulating substrate, an electrode formed on the substrate, an adhesive layer, an electrically insulating cover coupled to the substrate through the adhesive layer, and a sample transport path; A biosensor connecting sheet comprising two or more,
An electrically insulating sheet-like substrate whose longitudinal direction is the X-axis direction and the short direction is the Y-axis direction, and an electrically insulating sheet-like cover whose longitudinal direction is the X-axis direction and whose transverse direction is the Y-axis direction And an electrode formed on the substrate, an adhesive layer, and a sample transport path,
At least one set of positive and negative electrodes parallel to or orthogonal to the X-axis direction is opposed to the surface on the adhesive surface side of the sheet-like substrate,
An adhesive layer is formed on the adhesive surface side of the sheet-like substrate or the adhesive surface side of the sheet-like cover, and the sample transport groove patterned by the adhesive layer is oriented in a direction perpendicular to the electrodes. At least one formed,
There are two or more of at least one of the set of electrodes or the sample transport groove,
The biosensor connecting sheet, wherein the X-axis direction of the sheet-like substrate and the X-axis direction of the sheet-like cover coincide with each other, and the electrode and the sample transport groove intersect each other. The bio according to claim 15, wherein each electrode formed in the X-axis direction and / or each sample transport groove formed in the X-axis direction are continuously formed in the X-axis direction. Sensor connection sheet. In the vicinity of each set of electrodes or a part or all of the electrode surface, there is a reaction detection part further coated with a reagent layer,
The X-axis direction of the sheet-like substrate coincides with the X-axis direction of the sheet-like cover, the electrode and the sample transport groove intersect, and the reagent layer and the sample transport groove overlap each other. The biosensor connecting sheet according to claim 15 or 16, characterized in that

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