WO2013073073A1

WO2013073073A1 - Biosensor and biosensor production method

Info

Publication number: WO2013073073A1
Application number: PCT/JP2012/004441
Authority: WO
Inventors: 秀明大江; 高木　純; 憲二横山; 淳典平塚; 吉田　信行; 典子佐々木
Original assignee: 株式会社村田製作所
Priority date: 2011-11-18
Filing date: 2012-07-10
Publication date: 2013-05-23
Also published as: JPWO2013073073A1

Abstract

Provided is a technology that prevents a working electrode and a counter electrode from peeling off an insulating substrate, upon which an electrode layer is formed. In an area exposed by a cavity (104) in a conductive layer (112), notches (113a) are formed in the conductive layer (112) in such a manner as to cross the cavity (104) between one end of a working electrode (101), a counter electrode (102) and a sensing electrode (103), which are exposed by the cavity (104), and a sample introduction port (104a). Consequently, even if the conductive layer (112) in the area of a slit (105) where a spacer layer (120) is not laminated peels off the edge on the sample introduction port (104a) side of an electrode layer (110) (insulating substrate (111)) inside the cavity (104), there is no risk of the conductive layer (112) peeling beyond the notches (113a) formed in such a manner as to cross the cavity (104); thus, one end of the working electrode (101), the counter electrode (102) and the sensing electrode (103), which are exposed by the cavity (104), can be prevented from peeling off the electrode layer (110).

Description

Biosensor and biosensor manufacturing method

The present invention includes an electrode layer in which an electrode system including a working electrode and a counter electrode is provided on an insulating substrate, a reaction layer provided on the working electrode and the counter electrode, and a slit for forming a cavity to which a sample is supplied. The present invention relates to a biosensor including a formed spacer layer and a cover layer in which air holes communicating with a cavity are formed, and a method for manufacturing the biosensor.

As shown in an example of the conventional biosensor in FIG. 7, a biosensor having an electrode system including a working electrode 501 and a counter electrode 502 and a detection electrode 503, and a reaction layer 504 including an enzyme that specifically reacts with a measurement target substance. Oxidation obtained by applying a voltage between the working electrode 501 and the counter electrode 502 to oxidize the reducing substance produced by the reaction between the measurement target substance contained in the sample and the reaction layer 504 using 500. A method for measuring a substance that quantifies a substance to be measured by measuring an electric current is known (see, for example, Patent Document 1).

A biosensor 500 shown in FIG. 7 is a sensor for quantifying glucose contained in a sample, and includes an electrode layer 505 formed by providing an electrode on an insulating substrate such as polyethylene terephthalate or polyimide, and a cover layer. 507 and a spacer layer 506 disposed between the electrode layer 505 and the cover layer 507 are stacked. In addition, the spacer layer 506 is provided with a slit 506a for forming a cavity 508 to which a sample is supplied, and a cover layer 507 is laminated and bonded to the electrode layer 505 via the spacer layer 506. A cavity 508 to which a sample is supplied is formed by the electrode layer 505, the slit 506a portion of the spacer layer 506, and the cover layer 507. Then, a sample is supplied to the cavity 508 from the sample inlet formed on the side surface of the biosensor 500 by the opening portion of the slit 506a. The cover layer 507 is formed with an air hole 507a communicating with the terminal end of the cavity 508 in order to smoothly supply the sample to the cavity 508 by capillary action.

The electrode layer 505 is provided with a working electrode 501, a counter electrode 502, and a detection electrode 503, and an electrode pattern electrically connected to each of the electrodes 501 to 503 is provided, whereby an electrode system is provided in the electrode layer 505. Is formed. A reaction layer 504 is provided on the working electrode 501 and the counter electrode 502, and the working electrode 501, the counter electrode 502, and the detection electrode 503 are each exposed to a cavity 508 formed in the biosensor 500. Layer 505 is provided.

Therefore, when a sample made of liquid is supplied to the cavity 508 from the sample introduction port, the electrodes 501 to 503 exposed to the cavity 508 and the reaction layer 504 come into contact with the sample, and the reaction layer 504 is dissolved in the sample. Further, when the sample contacts the detection electrode 503, it is detected that the sample is supplied to the cavity 508.

The reaction layer 504 provided on the working electrode 501 and the counter electrode 502 includes glucose oxidase that specifically reacts with glucose contained in the sample and potassium ferricyanide as a mediator (electron acceptor). . The ferricyanide ions produced by the dissolution of potassium ferricyanide in the sample are reduced to ferrocyanide ions, which are reduced, by electrons released when glucose is oxidized to gluconolactone by reacting with glucose oxidase. The Accordingly, when a sample containing glucose is supplied to the cavity 508 formed in the biosensor 500 from the sample introduction port, ferricyanide ions are reduced by electrons released by the oxidation of glucose. Ferrocyanide ions, which are reduced forms of ferricyanide ions, are generated in an amount corresponding to the concentration of glucose contained and oxidized by the enzymatic reaction.

In the biosensor 500 configured as described above, the oxidation current obtained by oxidizing the reduced form of the mediator resulting from the enzyme reaction on the working electrode 501 has a magnitude depending on the glucose concentration in the sample. By measuring the current, glucose contained in the sample can be quantified.

Japanese Unexamined Patent Publication No. 2001-305093 (paragraphs 0016 to 0025, FIG. 1, etc.)

Incidentally, the electrode layer 505 is formed by providing an electrode system including a working electrode 501, a counter electrode 502, and a detection electrode 503 on an insulating substrate such as polyethylene terephthalate, polycarbonate, or polyimide. That is, the electrode layer 505 has a line-shaped cut 505b formed by laser processing on a conductive layer 505a formed of a noble metal such as gold, platinum, palladium, or a conductive material such as carbon over the entire surface of the insulating substrate. It is formed by providing an electrode system.

Therefore, the conductive layer 505a is provided up to the edge of the electrode layer 505 (biosensor 500). Therefore, in the conductive layer 505a provided on the electrode layer 505, the portion of the slit 506a in which the spacer layer 506 is not stacked and the conductive layer 505a at the rear end portion of the biosensor 500 inserted into the measuring instrument are When the sensor 500 is gripped or when the biosensor 500 is taken out from the wrapping paper, there is a possibility that the contact frictional force generated between the biosensor 500 and the contact object may peel off from the edge of the electrode layer 500.

In addition, the biosensor 500 generally includes a spacer layer 506 and a cover layer 507 formed on a large-area insulating substrate in which a plurality of electrode systems are provided by forming a line-shaped cut 505b in the conductive layer 505a. Are formed by stacking the biosensors 500 into individual pieces with a cutter. Therefore, when the assembly of biosensors 500 is separated into pieces, it is inserted into a slit 506a portion where the spacer layer 506 is not laminated or a measuring instrument due to stress generated when the insulating substrate is cut with a cutter. In the rear end portion of the biosensor 500, the conductive layer 500a may be peeled off from the edge of the electrode layer 500 cut by the cutter.

The present invention has been made in view of the above problems, and an object of the present invention is to provide a technique capable of preventing the working electrode and the counter electrode from being peeled off from the insulating substrate forming the electrode layer.

In order to achieve the above-described object, the biosensor of the present invention includes an electrode layer provided with an electrode system including a working electrode and a counter electrode on one surface of an insulating substrate, a slit, and the slit serving as the function. A spacer layer disposed on one end of the electrode and the counter electrode and stacked on the one surface of the electrode layer; a cavity formed by the electrode layer and the slit and supplied with a sample; and air communicating with the cavity A cover layer that is formed with a hole to cover the cavity and is laminated on the spacer layer, a sample introduction port that is formed by an opening of the slit and communicates with the cavity, the working electrode that is exposed to the cavity, and In a biosensor comprising a reaction layer provided on one end of the counter electrode, the electrode system is provided over the entire surface of one surface of the electrode layer. The cavity is formed between the one end side of the working electrode and the counter electrode and the sample introduction port at a portion of the conductive layer exposed to the cavity by forming a line-shaped cut in the conductive layer. The notch is formed so as to cross the line (Claim 1).

The spacer layer is laminated so as to partially cover the one surface of the electrode layer, and the other end side of each of the working electrode and the counter electrode is the electrode layer opposite to the sample introduction port. Of the edge portion of the electrode layer that is formed to extend to the edge of the electrode layer on which the spacer layer is not laminated, and on which the other end side of the working electrode and the counter electrode is formed. In the conductive layer, the cut is further formed so as to cross the other end side of each of the working electrode and the counter electrode (Claim 2).

Further, the biosensor manufacturing method of the present invention includes an electrode layer provided with an electrode system including a working electrode and a counter electrode on one surface of an insulating substrate, a slit, and the slit serving as the working electrode and the counter electrode. A spacer layer disposed on one end of the electrode layer and stacked on the one surface of the electrode layer, a cavity formed by the electrode layer and the slit and supplied with a sample, and an air hole communicating with the cavity. A cover layer that covers the cavity and is laminated on the spacer layer, a sample introduction port that is formed by the opening of the slit and communicates with the cavity, and one end of the working electrode and the counter electrode that are exposed to the cavity In the manufacturing method of a biosensor comprising a reaction layer provided on the side, the electrode layer is a conductive layer over the entire surface of one side of the insulating substrate. When the electrode system is formed by forming a line-shaped notch in the conductive layer, a portion of the conductive layer exposed to the cavity is exposed to one end side of the front working electrode and the counter electrode, and the sample introduction The notch is prepared so as to cross the cavity between the mouth and the mouth (claim 3).

The biosensor manufacturing method according to claim 4, wherein the spacer layer is laminated so as to partially cover one surface of the electrode layer, and the electrode layer includes the working electrode and the counter electrode, respectively. An end side is formed to extend to an end edge of the electrode layer on the side opposite to the sample introduction port and the spacer layer is not laminated, and the working electrode and the counter electrode The conductive layer at the edge portion of the electrode layer on which the other end side is formed is prepared by further forming the cut so as to cross the other end side of each of the working electrode and the counter electrode. (Claim 4).

Further, the cut is formed in the conductive layer by laser processing (Claim 5).

According to the first and third aspects of the invention, the electrode system including the working electrode and the counter electrode provided on one surface of the insulating substrate forming the electrode layer is provided over the entire surface of the one surface of the insulating substrate. It is formed by forming a line-shaped cut in the conductive layer. In the portion of the conductive layer exposed to the cavity, a cut is formed in the conductive layer so as to cross the cavity between one end side of the working electrode and the counter electrode exposed to the cavity and the sample introduction port.

Therefore, even if the conductive layer in the slit portion where the spacer layer is not stacked is peeled off from the edge of the insulating substrate on the sample inlet side in the cavity, the conductive layer is formed so as to cross the cavity. Therefore, the working electrode and the one end of the counter electrode exposed to the cavity can be prevented from being peeled off from the insulating substrate.

According to the second and fourth aspects of the present invention, the spacer layer is laminated so as to partially cover one surface of the electrode layer, and the other end side of each of the working electrode and the counter electrode is opposite to the sample introduction port. The electrode layer is formed to extend to the edge of the electrode layer on which the spacer layer is not stacked. A cut is formed in the conductive layer at the edge portion of the electrode layer where the other end side of the working electrode and the counter electrode is formed so as to cross the other end side of each of the working electrode and the counter electrode.

Therefore, even if the conductive layer forming the other end of each of the working electrode and the counter electrode on which the spacer layer is not laminated is peeled off from the edge of the insulating substrate, the conductive layer is not connected to the other end of each of the working electrode and the counter electrode. Since there is no possibility of peeling beyond the cut formed so as to cross, it is possible to prevent the working electrode on which the spacer layer is not stacked and the other end of the counter electrode from being peeled off from the insulating substrate.

According to the invention of claim 5, it is possible to easily and accurately form the cut in the conductive layer by laser processing.

It is a top view of the insulating substrate in a certain manufacturing process of the biosensor concerning one Embodiment of this invention. It is a top view which shows the state in which the conductive layer was provided over the whole one surface of the insulating board | substrate in the manufacturing process from which the biosensor concerning one Embodiment of this invention differs. It is a figure which shows the further different manufacturing process of the biosensor concerning one Embodiment of this invention, Comprising: It is a principal part enlarged view which shows the state by which the cut was formed in the conductive layer. It is a figure which shows the further different manufacturing process of the biosensor concerning one Embodiment of this invention, Comprising: It is a principal part enlarged view which shows the state by which the spacer layer was laminated | stacked on the electrode layer. It is a figure which shows the further different manufacturing process of the biosensor concerning one Embodiment of this invention, Comprising: It is a principal part enlarged view which shows the state by which the reaction layer was provided in the cavity. It is a top view which shows the biosensor concerning one Embodiment of this invention. It is a figure which shows the conventional biosensor.

A configuration of a biosensor according to an embodiment of the present invention and a manufacturing method thereof will be described with reference to FIGS.

FIGS. 1 to 5 are diagrams showing a method of manufacturing a biosensor according to an embodiment of the present invention, in which FIG. 1 is a plan view of an insulating substrate and FIG. 2 is an entire surface of one surface of the insulating substrate. 3 is a plan view showing a state where a conductive layer is provided, FIG. 3 is an enlarged view of a main part showing a state where a cut is formed in the conductive layer, and FIG. 4 is a main part showing a state where a spacer layer is laminated on the electrode layer. FIG. 5 is an enlarged view of a main part showing a state in which a reaction layer is provided in the cavity. FIG. 6 is a plan view of the biosensor. 1 and FIG. 2 indicates a cutting line CL when the biosensor 100 is separated. FIGS. 3 to 5 are enlarged views of essential parts showing regions surrounded by broken lines in FIGS.

The biosensor 100 of the present invention has substantially the same configuration as the conventional biosensor 500 described with reference to FIG. 7, and includes an electrode system including a working electrode 101, a counter electrode 102, and a detection electrode 103, and measurement. It has a reaction layer 107 containing an enzyme that reacts with a target substance, and is used by being attached to a measuring instrument (not shown). That is, a substance to be measured such as glucose contained in a sample such as blood supplied to a cavity 104 provided on the distal end side of the biosensor 100 attached to the measuring instrument, and a reaction layer 107 provided on the biosensor 100 By measuring the oxidation current obtained by applying a voltage between the working electrode 101 and the counter electrode 102 to oxidize the reducing substance produced by the reaction of the reaction, the quantitative determination of the measurement target substance contained in the sample Is done.

That is, as shown in FIGS. 3 to 6, the biosensor 100 includes a working electrode 101 and a counter electrode 102 formed of an insulating material such as ceramic, glass, plastic, paper, biodegradable material, and polyethylene terephthalate, respectively. In addition, an electrode layer 110 provided with an electrode system including the detection electrode 103, a cover layer 130 in which an air hole 106 communicating with the cavity 104 is formed, and a slit 105 for forming the cavity 104 are formed. 110 and the spacer layer 120 disposed between the cover layers 130 are laminated and bonded together with the tip side where the sample inlet 104a is provided aligned. Further, as shown in FIG. 5, a reaction layer 107 including an enzyme that reacts with a measurement target substance such as glucose contained in a sample is provided on the working electrode 101 and the counter electrode 102. Then, the biosensor 100 is attached to the measuring device 2 by being inserted and attached to a predetermined insertion port of the measuring device 2 from the rear end side.

In this embodiment, the electrode layer 110 is prepared as follows.

That is, first, as shown in FIGS. 1 and 2, on one surface of an insulating substrate 111 made of an insulating material such as polyethylene terephthalate, noble metals such as platinum, gold, and palladium are formed by screen printing or sputtering deposition. A conductive layer 112 made of a conductive material such as carbon, copper, aluminum, titanium, ITO, or ZnO is formed. Next, as shown in FIG. 3, the conductive layer 112 is patterned by laser processing or photolithography to form a cut 113, thereby including the working electrode 101, the counter electrode 102, and the detection electrode 103. An electrode system is formed.

4 and FIG. 5, the working electrode 101, the counter electrode 102, and the detection electrode 103 are arranged so that one end sides thereof are exposed to the cavity 104. Further, the other end side of each of the working electrode 101, the counter electrode 102, and the detection electrode 103 is an edge of the electrode layer 110 on the side opposite to the sample introduction port 104a, and is an end of the electrode layer 110 on which the spacer layer 120 is not laminated. It is stretched to the edge.

At this time, as shown in FIGS. 3 to 5, the conductive layer 112 includes a working electrode 101, a counter electrode 102, and a detection electrode in a portion exposed to the cavity 104 when the spacer layer 120 is laminated on the electrode layer 110. A notch 113a is formed between one end side of 103 and the sample introduction port 104a so as to cross the cavity 104. In addition, the conductive layer 112 at the edge of the electrode layer 110 on which the other end side of the working electrode 101 and the counter electrode 102 and the detection electrode 103 is formed has the other end of each of the working electrode 101, the counter electrode 102 and the detection electrode 103. A cut 113b is formed so as to cross the side.

As shown in FIGS. 1 and 2, a plurality of electrode systems for forming the biosensor 100 are formed on one surface of the insulating substrate 111 and cut after the cover layer 130 is laminated as will be described later. By being cut along the line CL, the individual biosensors 110 are separated. Therefore, the distance from the front end of the electrode layer 110 to the notch 113a and the distance from the rear end of the electrode layer 110 to the notch 113b are the cutting position accuracy when the assembly of the biosensor 100 is cut along the cutting line CL. It is better to make it larger.

Next, the spacer layer 120 is laminated on the electrode layer 110 prepared as described above. As shown in FIGS. 4 and 5, the spacer layer 120 is formed of a substrate made of an insulating material such as polyethylene terephthalate, and a slit 105 for forming the cavity 104 is formed in the approximate center of the front edge of the substrate. ing. The slit 105 is disposed on one end side of the working electrode 101, the counter electrode 102, and the detection electrode 103, and the spacer layer 120 is partially covered and laminated on one surface of the electrode layer 110, whereby the electrode layer 110 is stacked. A cavity 104 to which a sample is supplied is formed by the slit 105.

Subsequently, the reaction layer 107 is formed after the cavity 104 formed by laminating the spacer layer 120 on the electrode layer 110 is cleaned with plasma. As the plasma used in the plasma cleaning process, various plasmas used in metal activation treatment by plasma such as oxygen plasma, nitrogen plasma, and argon plasma can be used. Plasma may be used.

As shown in FIG. 5, before the cover layer 130 is laminated on the spacer layer 120, the reaction layer 107 is formed on one end side of the working electrode 101 and the counter electrode 102 exposed to the cavity 104 and the detection electrode 103 with carboxymethylcellulose or the like. It is formed by dropping a reagent containing a thickener such as gelatin, an enzyme, a mediator, an additive such as an amino acid or an organic acid. In addition, a hydrophilic agent such as a surfactant or phospholipid is applied to the inner wall of the cavity 104 in order to smoothly supply a sample such as blood to the cavity 104.

Enzymes include glucose oxidase, lactate oxidase, cholesterol oxidase, alcohol oxidase, sarcosine oxidase, fructosylamine oxidase, pyruvate oxidase, glucose dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, hydroxybutyrate dehydrogenase, cholesterol esterase, creatininase, creatinase DNA polymerase or the like can be used, and various sensors can be formed by selecting these enzymes according to the substance to be measured.

For example, glucose oxidase or glucose dehydrogenase can be used to form a glucose sensor that detects glucose in a blood sample, and alcohol oxidase or alcohol dehydrogenase can be used to form an alcohol sensor that detects ethanol in a blood sample. For example, a lactic acid sensor for detecting lactic acid in a blood sample can be formed, and a total cholesterol sensor can be formed by using a mixture of cholesterol esterase and cholesterol oxidase.

As the mediator, potassium ferricyanide, ferrocene, ferrocene derivatives, benzoquinone, quinone derivatives, osmium complexes, ruthenium complexes and the like can be used.

As the thickener, carboxymethyl cellulose, carboxyethyl cellulose, polyethyleneimine DEAE cellulose dimethylaminoethyl dextran carrageenan sodium alginate dextran and the like can be used. As the hydrophilizing agent, surfactants such as Triton X100 (manufactured by Sigma Aldrich), Tween 20 (manufactured by Tokyo Chemical Industry Co., Ltd.), sodium bis (2-ethylhexyl) sulfosuccinate, and phospholipids such as lecithin can be used.

Also, a buffer such as phosphoric acid may be provided in order to reduce the variation in the concentration of ions contained in the sample.

Next, after the reaction layer 107 is formed in the cavity 104, a cover layer 130 formed of a substrate made of an insulating material such as polyethylene terephthalate is laminated on the spacer layer 120. As shown in FIG. 6, the cover layer 130 has an air hole 106 communicating with the cavity 104 when laminated on the spacer layer 120. The cover layer 130 covers the cavity 104 and covers the spacer layer 120. Is laminated.

Then, the assembly of biosensors 110 formed by laminating the cover layer 130 is cut along the cutting line CL, and the biosensor 100 is separated into individual pieces, thereby forming the openings of the slits 105. Thus, the biosensor 100 having the sample introduction port 104 a communicating with the cavity 104 at the tip is formed.

In this embodiment, the biosensor 100 is formed for the purpose of quantifying glucose in blood, and FAD (flavin adenine dinucleotide) is used as an enzyme that specifically reacts with glucose as a measurement target substance. A mediator that contains GDH (glucose dehydrogenase) (hereinafter referred to as FAD-GDH), which is reduced by electrons generated by the reaction of glucose as a measurement object and FAD-GDH, and becomes a reducing substance The reaction layer 107 containing potassium ferricyanide is provided on one end side of the working electrode 101 and the counter electrode 102 exposed to the cavity 104.

In the biosensor 100 configured as described above, by bringing a sample made of blood into contact with the sample introduction port 104a at the tip, the sample is sucked toward the air hole 106 by capillary action, and the sample is supplied to the cavity 104. . Then, when the reaction layer 107 is dissolved in the sample supplied to the cavity 104, electrons are released by an enzymatic reaction between glucose and FAD-GDH as a measurement target substance in the sample, and ferricyanization is performed by the emitted electrons. The ions are reduced to produce ferrocyanide ions, which are reducing substances. Then, by applying a voltage between the working electrode 101 and the counter electrode 102 of the biosensor 100 to electrochemically oxidize the reducing substance generated by the redox reaction caused by the reaction layer 107 dissolving in the sample. By measuring the oxidation current flowing between the working electrode 101 and the counter electrode 102, glucose in the sample is quantified in the measuring device.

As described above, according to this embodiment, the electrode system including the working electrode 101, the counter electrode 102, and the detection electrode 103 provided on one surface of the insulating substrate 111 that forms the electrode layer 110 is the insulating substrate 111. This is formed by forming a line-shaped cut 113 in the conductive layer 112 provided over the entire surface of one side. Then, in the portion exposed to the cavity 104 of the conductive layer 112, the conductive electrode 101 and the counter electrode 102 exposed to the cavity 104 and the one end side of the detection electrode 103 and the sample introduction port 104 a are traversed across the cavity 104. A cut 103 a is formed in the layer 112.

The spacer layer 120 is laminated so as to partially cover one surface of the electrode layer 110, and the other end side of each of the working electrode 101, the counter electrode 102, and the detection electrode 103 is opposite to the sample introduction port 104a. The electrode layer 110 is extended to the edge of the electrode layer 110 on which the spacer layer 120 is not stacked. The conductive layer 112 at the edge of the electrode layer 110 on which the other end side of the working electrode 101 and the counter electrode 102 and the detection electrode 103 is formed has the other end of each of the working electrode 101, the counter electrode 102 and the detection electrode 103. A cut 103b is formed so as to cross the side.

Therefore, when the aggregate of the biosensors 100 is separated into pieces, when the insulating substrate 111 is cut with a cutter along the cutting line CL, or when the biosensor 500 is gripped, the biosensor 500 is packaged. When taken out from paper, the conductive layer 112 in the portion of the slit 105 where the spacer layer 120 is not stacked is formed in the cavity 104 from the edge of the electrode layer 110 (insulating substrate 111) on the sample inlet 104a side. Even if the conductive layer 112 is peeled off, the conductive layer 112 is not likely to peel off beyond the notch 113 a formed so as to cross the cavity 104, so that one end side of the working electrode 101 and the counter electrode 102 and the detection electrode 103 exposed to the cavity 104 is The peeling from the electrode layer 110 can be prevented.

Similarly, the conductive layer 112 that forms the other end of each of the working electrode 101 and the counter electrode 102 and the detection electrode 103 on which the spacer layer 120 is not laminated is the edge of the electrode layer 110 (insulating substrate 111). The conductive layer 112 is not likely to be peeled beyond the notch 103b formed so as to cross the other end of each of the working electrode 101, the counter electrode 102, and the detection electrode 103 even if the spacer layer 120 is peeled off. It is possible to prevent the other end side of the working electrode 101 and the counter electrode 102 and the detection electrode 103 that are not stacked from being peeled off from the electrode layer 110.

Further, the

cuts

113, 113a, 113b can be formed in the conductive layer 112 by laser processing or photolithography as described above. In particular, the

cuts

113, 113a, 113b are formed in the conductive layer 112 by laser processing. In this case, the

cuts

113, 113a, 113b can be formed in the conductive layer 112 at low cost and with high accuracy.

Note that the present invention is not limited to the above-described embodiment, and various modifications other than those described above can be made without departing from the spirit thereof. For example, the reaction of the above-described biosensor 100 is possible. An ethanol sensor, a lactic acid sensor, or the like may be formed by changing the combination of the enzyme and the mediator included in the layer 107. In addition, the reaction layer 107 does not necessarily include a mediator. In this case, an oxidation current generated by oxidation of a reducing substance such as hydrogen peroxide or a reduced form of the enzyme generated by an enzyme reaction of a measurement target substance such as glucose is generated. Just measure.

In the above-described embodiment, the biosensor 100 is formed in a bipolar electrode structure having the working electrode 101 and the counter electrode 102. However, the biosensor 100 is formed in a tripolar electrode structure by further providing a reference electrode. May be. In this case, a predetermined potential based on the counter electrode 102 may be applied to the working electrode 101 in a state where the counter electrode 102 is grounded and a reference potential is applied to the reference electrode by the voltage output unit.

Further, in the above-described embodiment, by applying a predetermined voltage between the counter electrode 102 and the detection electrode 103, the current flowing between the counter electrode 102 and the detection electrode 103 is monitored, thereby allowing blood to enter the cavity 104. Although it is configured to detect that the sample has been supplied, the detection electrode 103 is not necessarily provided. In this case, by applying a predetermined voltage between the working electrode 101 and the counter electrode 102, the current flowing between the working electrode 101 and the counter electrode 102 is monitored to confirm that the blood sample has been supplied to the cavity 104. What is necessary is just to detect.

In addition, of the electrode layer 110, the spacer layer 120, and the cover layer 130 that form the biosensor 100, at least the cover layer 130 is formed of a transparent member so that it can be visually recognized that the blood sample is supplied to the cavity 104. Is desirable.

The present invention can be applied to various biosensors and methods for manufacturing the biosensors.

DESCRIPTION OF SYMBOLS 100 Biosensor 101 Working electrode 102 Counter electrode 104 Cavity 104a Sample inlet 105 Slit 106 Air hole 107 Reaction layer 110 Electrode layer 111 Insulating substrate 112

Conductive layer

113a, 113b Cut 120 Spacer layer 130 Cover layer

Claims

An electrode layer provided with an electrode system including a working electrode and a counter electrode on one surface of an insulating substrate;
A spacer layer formed on the one surface of the electrode layer, the slit being disposed on one end side of the working electrode and the counter electrode;
A cavity formed by the electrode layer and the slit and supplied with a sample;
An air hole communicating with the cavity to cover the cavity and be laminated on the spacer layer;
A sample introduction port formed by the opening portion of the slit and communicating with the cavity;
In a biosensor comprising the working electrode exposed in the cavity and a reaction layer provided on one end side of the counter electrode,
The electrode system is formed by forming a line-shaped cut in a conductive layer provided over the entire surface of one side of the electrode layer,
In the portion of the conductive layer exposed to the cavity, the cut is formed so as to cross the cavity between one end side of the working electrode and the counter electrode and the sample introduction port. Biosensor.
The spacer layer is laminated so as to partially cover the one surface of the electrode layer,
The other end side of each of the working electrode and the counter electrode is an edge of the electrode layer opposite to the sample introduction port, and is extended to the edge of the electrode layer on which the spacer layer is not stacked. In the conductive layer at the edge portion of the electrode layer on which the other end side of the working electrode and the counter electrode is formed, the cut is made so as to cross the other end side of each of the working electrode and the counter electrode. The biosensor according to claim 1, further formed.
An electrode layer provided with an electrode system including a working electrode and a counter electrode on one surface of an insulating substrate;
A spacer layer formed on the one surface of the electrode layer, the slit being disposed on one end side of the working electrode and the counter electrode;
A cavity formed by the electrode layer and the slit and supplied with a sample;
An air hole communicating with the cavity to cover the cavity and be laminated on the spacer layer;
A sample introduction port formed by the opening portion of the slit and communicating with the cavity;
In a manufacturing method of a biosensor comprising the working electrode exposed in the cavity and a reaction layer provided on one end side of the counter electrode,
The electrode layer is
A portion of the conductive layer exposed to the cavity when forming the electrode system by forming a conductive layer over the entire surface of one side of the insulating substrate and forming a line-shaped cut in the conductive layer. A method of manufacturing a biosensor, comprising: preparing a notch so as to cross the cavity between one end side of the front working electrode and the counter electrode and the sample introduction port.
The biosensor manufacturing method according to claim 3, wherein the spacer layer is partially covered and laminated on one surface of the electrode layer.
The electrode layer is
The other end side of each of the working electrode and the counter electrode is formed by extending to the edge of the electrode layer on the side opposite to the sample introduction port, where the spacer layer is not stacked. And further cutting the cut into the conductive layer at the edge portion of the electrode layer where the other end of the working electrode and the counter electrode is formed so as to cross the other end of each of the working electrode and the counter electrode. A method for manufacturing a biosensor, characterized by being prepared.
The biosensor manufacturing method according to claim 4, wherein the cut is formed in the conductive layer by laser processing.