WO2009133983A1 - Biodétecteur - Google Patents

Biodétecteur Download PDF

Info

Publication number
WO2009133983A1
WO2009133983A1 PCT/KR2008/002862 KR2008002862W WO2009133983A1 WO 2009133983 A1 WO2009133983 A1 WO 2009133983A1 KR 2008002862 W KR2008002862 W KR 2008002862W WO 2009133983 A1 WO2009133983 A1 WO 2009133983A1
Authority
WO
WIPO (PCT)
Prior art keywords
layer
sample
conductive
conductive coupling
coupling layer
Prior art date
Application number
PCT/KR2008/002862
Other languages
English (en)
Inventor
Keum Pil Lee
Original Assignee
Keum Pil Lee
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Keum Pil Lee filed Critical Keum Pil Lee
Publication of WO2009133983A1 publication Critical patent/WO2009133983A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
    • G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels

Definitions

  • the present invention relates to a biosensor, and more particularly, to a biosensor, in which a coupling layer for coupling an upper substrate to a lower substrate, either of which is provided with a working electrode, includes a conductive material and is thus used as a counter electrode or a sample recognition electrode .
  • US Patent No. 5,437,999 discloses a sensor formed by patterning a working electrode or a counter electrode on an insulating substrate using photolithography to precisely define an electrode area and then coupling the substrate having the above electrode to another substrate to face each other by an intermediate coupling layer disposed therebetween.
  • One side of the intermediate coupling layer is formed with an empty space, into which a measurement sample is injected and which includes a reagent reactive with an analyte in the sample, and further, the substrate having the working electrode or counter electrode is provided with an air vent for discharging air.
  • US Patent No. 5,582,697 discloses a biosensor for quantifying an enzyme substrate present in a sample through an electrochemical method.
  • the biosensor is composed of a working electrode, a counter electrode, and a third electrode, which are formed on an insulating substrate, and further, a reaction layer containing oxidoreductase is formed over the working electrode and the counter electrode.
  • the third electrode is used as an electrode for sensing the injection of a sample, and is located farther away than the working electrode and counter electrode from a sample injection part.
  • US Patent Nos. 6,071,391 and 6,156,173 disclose a biosensor, in which an upper substrate is coupled to a lower substrate by means of an intermediate coupling layer disposed therebetween, thus forming a sample injection space, and a working electrode and a counter electrode are formed on the two substrates, respectively, to face each other. These two substrates lack an air vent, and the sample injection part of each of the substrates provided with the working electrode and counter electrode is tapered. Further, in order to connect the electrode formed on the upper substrate to a lead wire of the lower substrate, a portion of the coupling layer is bored, and is then filled with a conductive material.
  • US Patent No. 6,576,101 discloses a biosensor for electrochemically measuring a sample of 1 ⁇ l or less, in which a redox material that is air-oxidizable and is bound to a polymer is applied, together with a reactive enzyme, onto a working electrode.
  • the redox material bound to the polymer functions as an electron mediator upon reaction with the enzyme on the working electrode and is characterized in that it does not diffuse in the sample solution.
  • US Patent No. 6,618,934 discloses a method of manufacturing a plurality of electrochemical sensors, including forming pluralities of working electrodes and counter electrodes in a first electrode zone and a second electrode zone on a substrate, forming a spacer layer in either of the electrode zones, removing a portion of the spacer to form a sample reaction chamber, folding the substrate to layer the first electrode zone and the second electrode zone, thus manufacturing the electrochemical sensors, which are then cut into each sensor.
  • Each sensor is composed of one or more working electrodes, one or more counter electrodes, and the sample reaction chamber.
  • US Patent No. 6,863,800 discloses a biosensor for measuring an analyte, in which a spacer is disposed between an electrode formed on an electrode support and another electrode formed on a cover layer.
  • the working electrode is formed of working ink including a reagent reactive with the analyte, a conductive material, and an electron mediator, and the reference electrode is formed by applying a material reactive with the analyte and an electron mediator onto the conductive material.
  • the counter electrode is formed of a conductive material, and the electrode support and the cover layer are disposed to face each other.
  • US Patent No. 6,885,196 discloses a biosensor composed of a first insulating substrate having a working electrode and a second insulating substrate having a counter electrode, which are disposed to face each other, in which a sample supply pathway is formed between the two substrates, and the working electrode, the counter electrode, and a reaction layer containing oxidoreductase are exposed to the sample supply pathway. Further, the distance between the working electrode and the counter electrode is 150 ⁇ m or less, and the area of the counter electrode exposed to the sample supply pathway is smaller than the area of the working electrode exposed thereto.
  • US Patent No. ⁇ ,942,769B2 discloses a biosensor, in which an electrode is formed on an electrode support, and an insulating layer, an adhesive layer, a screen having porosity of 10-40%, another adhesive layer, and a cover layer are sequentially formed and assembled on the electrode support and the electrode.
  • the insulating layer and the cover layer have an open slot, and the screen functions to control the flow and volume of the analyte into the slot formed in the insulating layer and the cover layer.
  • US Patent No. 7,022,218 discloses a biosensor for analyzing a very small amount of sample, in which a working electrode having a plurality of branches and a first counter electrode having a plurality of branches are alternately arranged on a first insulating substrate, and a second counter electrode having branches are formed on a second insulating substrate.
  • the two substrates are disposed to face each other, and a sample supply pathway, which is defined between the two substrates, includes a reactive reagent containing oxidoreductase and the working electrode and the counter electrode which are alternately arranged.
  • 7,050,843 discloses a biosensor, in which a first insulating substrate having a conductive surface and a second substrate having a conductive surface are coupled to face each other by means of an intermediate layer disposed therebetween, and a capillary channel defined by the two substrates and the intermediate layer includes a reagent in a dry form that reacts with a sample.
  • an insulating pattern line is provided on the conductive surface, so that the flow of the sample is controlled and the conductive surface is divided into two sections.
  • US Patent Application Publication No. 2006/0008581 Al discloses a method of manufacturing an electrochemical sensor, including forming a working electrode on a first insulating material, forming multiple insulating layers on the working electrode, creating a hole through the working electrode, thus forming a receptacle, the cut surface of the working electrode being exposed to the wall of the receptacle, forming a reference electrode on the uppermost insulating layer among the multiple insulating layers, and coupling a lower substrate.
  • the biosensor for electrochemically measuring the analyte is composed basically of an upper insulating substrate, a lower insulating substrate, and an intermediate layer for coupling the above two substrates, and a predetermined space for sample injection or sample reaction is defined by the above two substrates and the intermediate layer.
  • the two substrates are provided with the working electrode and the counter electrode for measuring electrochemical oxidation or reduction current.
  • the area of the working electrode and counter electrode formed on the substrate, coming into contact with the sample is regarded as important.
  • electrochemical signals are increased in proportion to the area of the electrode.
  • the above patents are characterized in that one of the two substrates is provided with the working electrode, and the other substrate is formed with the counter electrode, and these two electrodes are disposed to face each other using an intermediate layer.
  • Such an electrode arrangement is advantageous because the electrode area may be more efficiently maximized than when the two electrodes are formed on the same substrate.
  • the conductive material used for almost all of the electrodes is opaque.
  • the sample injection is difficult to observe with the naked eye.
  • the present invention has been made keeping in mind the above problems occurring in the prior art, and the present invention provides a biosensor, in which a coupling layer for coupling an upper substrate and a lower substrate, either of which is provided with a working electrode, includes a conductive material and is thus used as a counter electrode or a sample recognition electrode .
  • a biosensor for measuring an analyte in a sample may comprise at least one working electrode, which is formed on either or both of an upper substrate and an insulating substrate and is used to measure signals generated by the analyte; and at least one conductive coupling layer, which is used to couple said two substrates to each other and has electrical conductivity; wherein at least one reaction chamber for reaction of the analyte is defined by the coupled two substrates and the conductive coupling layer and wherein one side surface of the conductive coupling layer exposed to the reaction chamber is used as a counter electrode for measuring signals generated by the analyte .
  • the working electrode may comprise at least any one selected from among a conductive polymer, gold, palladium, graphite, carbon, ITO particles, metal particles, and carbon nanotubes (CNTs) . Further, one side surface of the conductive coupling layer may be irregularly formed to increase a contact area with the sample.
  • the conductive coupling layer may be formed by printing or coating both surfaces of an insulating layer with a conductive adhesive material.
  • the conductive coupling layer may be formed by applying a conductive adhesive material onto one surface of an insulating layer and applying a non-conductive adhesive material onto the other surface of the insulating layer.
  • the conductive coupling layer may be formed exclusively of a conductive adhesive material.
  • the conductive coupling layer may comprise at least any one selected from among a conductive polymer, gold, silver, silver chloride, palladium, graphite, carbon, ITO particles, metal particles, and CNTs.
  • the coupling of the upper substrate, the insulating substrate and the conductive coupling layer may be realized using pressure, heat or light.
  • the reaction chamber may be provided with a reaction layer comprising at least one enzyme, which reacts with the analyte, and an electron mediator, which reacts with the enzyme to thus be oxidizable or reducible.
  • the reaction layer is preferably formed in the reaction chamber and underneath the conductive coupling layer. Further, the reaction layer preferably comprises a water- soluble polymer.
  • the enzyme contained in the reaction layer is preferably at least any one selected from among oxidoreductase, transferase, hydrolase, lyase, isomerase, and synthetase.
  • the reaction layer preferably further comprises a buffer having a pH ranging from 2 to 9.
  • the conductive coupling layer may have a thickness ranging from 1 /M to 1000 ⁇ m.
  • the conductive coupling layer may have a sheet resistance ranging from 10 ⁇ to 500 k ⁇ .
  • the biosensor may further comprise a porous layer, which is adhered over the working electrode by the conductive coupling layer.
  • the porous layer preferably comprises micropores having a size of 10 /zm or less to remove erythrocytes in a sample.
  • the porous layer preferably contains an enzyme which reacts with the analyte in the sample.
  • the biosensor may further comprise a sample recognition electrode, which is formed with the conductive coupling layer in the reaction chamber defined by the upper substrate, the insulating substrate, and the conductive coupling layer and which functions to sense the supply of the sample into the reaction chamber.
  • the sample recognition electrode may be spaced apart from a sample injection part for injection of the sample so that it is located farther away than the working electrode.
  • the biosensor may further comprise an air vent which is connected to the reaction chamber defined by the upper substrate, the insulating substrate, and the conductive coupling layer.
  • a conductive coupling layer is used as a counter electrode or a sample recognition electrode, and thus, the need to additionally provide the above electrodes on the substrate is eliminated, thereby maximizing the area of a working electrode.
  • FIG. 1 is a view sequentially showing a process of manufacturing a biosensor according to a first embodiment of the present invention
  • FIG. 2 is a cross-sectional view showing the biosensor according to the first embodiment of the present invention.
  • FIG. 3 is a cross-sectional view showing the biosensor according to the first embodiment of the present invention, in which a reaction layer is formed in a reaction chamber;
  • FIG. 4 is a cross-sectional view showing the biosensor according to the first embodiment of the present invention, in which a reaction layer is formed in a reaction chamber and underneath portions of conductive coupling layers;
  • FIG. 5 is a cross-sectional view showing the conductive coupling layer of the biosensor according to the present invention.
  • FIG. 6 is a view sequentially showing a process of manufacturing a biosensor including a sample recognition electrode for sensing the supply of a sample into a reaction chamber, according to the present invention
  • FIG. 7 is a view sequentially showing a process of manufacturing a biosensor including a porous layer, which is adhered over a working electrode by a conductive coupling layer, according to the present invention
  • FIG. 8 is a view sequentially showing a process of manufacturing a biosensor including a sample injection part and a sample reaction chamber on both sides thereof, according to the present invention
  • FIG. 9 is a view sequentially showing a process of manufacturing a biosensor having a plurality of reaction chambers according to a second embodiment of the present invention.
  • FIG. 10 is a view sequentially showing a process of manufacturing a biosensor having a facing structure, according to a third embodiment of the present invention.
  • FIG. 11 is a cross-sectional view showing the biosensor having a facing structure, according to the third embodiment of the present invention.
  • FIG. 12 is a view sequentially showing a process of manufacturing a biosensor having a plurality of reaction chambers according to a fourth embodiment of the present invention ;
  • FIG. 13 is a cross-sectional view showing the biosensor having a plurality of reaction chambers according to the fourth embodiment of the present invention.
  • FIG. 14 is a view showing the biosensor according to the present invention, which is mounted in a rotatable rotator .
  • FIG. 1 sequentially shows a process of manufacturing a biosensor according to a first embodiment of the present invention.
  • conductive layers 20, 30 are formed on an insulating substrate 10.
  • the conductive layers may be formed through various methods, including printing, coating, deposition, etc.
  • an insulating layer 40 is formed in a pattern as shown in FIG. 1 (b) on the conductive layers 20, 30 and the insulating substrate 10, thus covering a portion of the conductive layer 20 which comes into contact with a conductive coupling layer 5OA which is to be formed in a subsequent process.
  • a working electrode 3OA and a connector 3OB for connecting the working electrode to a measurement system are formed, and a connector 2OA for electrical connection of the conductive layer 20 to a conductive coupling layer 5OB and a connector 2OB to a measurement system are formed.
  • FIG. 1 (c) illustrates a case where the conductive coupling layers 5OA, 5OB are used as a counter electrode, together with an exposure portion 2OC which is a portion of the conductive material coming into contact with the sample
  • FIG. l(c') illustrates a case where the conductive coupling layers 5OA, 5OB are used as a counter electrode in an electrochemical sensor.
  • the conductive coupling layer is formed of a material having electrical conductivity and adhesiveness, and is described later with reference to FIG. 5.
  • the insulating substrate 10 is coupled to an upper substrate 80 using adhesiveness of the conductive coupling layer disposed therebetween.
  • a sample injection part 90 for injection of a measurement sample a sample reaction chamber 70 for reaction of a measurement analyte, and an air vent 60 for discharge of air from the sample reaction chamber 70 into which the sample is injected from the sample injection part 90 are formed.
  • the upper substrate, the insulating substrate, and the conductive coupling layers are preferably coupled using pressure, heat or light.
  • the conductive coupling layers 5OA, 5OB are formed of a conductive material and are electrically connected with the conductive layer 20 via the connector 2OA.
  • FIG. 2 shows the cross-section of the biosensor according to the first embodiment of the present invention .
  • conductive coupling layers 5OC, 5OD which are exposed to the sample reaction chamber 70, are brought into contact with an analyte, and are thus used as a counter electrode or a reference electrode in the electrochemical sensor or as a sample recognition electrode for sensing the supply of the sample being inserted into the sample reaction chamber 70.
  • the use of the conductive coupling layers 5OA, 5OB, 5OC, 5OD as such electrodes is advantageous because a minimum space occupation is ensured in the limited sample reaction chamber 70, thus maintaining the maximum possible area of the working electrode 3OA.
  • the working electrode includes at least any one selected from among a conductive polymer, gold, palladium, graphite, carbon, ITO particles, metal particles, and CNTs.
  • FIG. 3 shows a cross-section of the biosensor according to the first embodiment of the present invention, in which a reaction layer is formed in the reaction chamber
  • FIG. 4 shows the cross-section of the biosensor according to the first embodiment of the present invention, in which a reaction layer is formed in the reaction chamber and underneath portions of the conductive coupling layers.
  • a reaction layer 500 for measurement of the analyte of the sample is formed in the sample reaction chamber 70 of the biosensor 100.
  • the reaction layer 500 may be formed on the working electrode 3OA, and portions 500A, 500B of the reaction layer 500 may be formed underneath the conductive coupling layers 5OA, 5OB.
  • the reaction layer 500 includes an enzyme, which reacts with the analyte to thus generate analytic signals able to be measured by the working electrode 3OA and the type of which varies depending on the analyte. Further, such a reaction layer 500 includes a polymer.
  • the reaction layer is dissolved or wet, or absorbs the sample, due to the supply of the sample, and is thus connected with the conductive coupling layers in contact therewith.
  • the contact area between the sample and the conductive coupling layer may be relatively enlarged via the reaction layer.
  • the polymer for the reaction layer 500, 500A, 500B includes a water-soluble polymer or a water-insoluble polymer, and preferably includes a water- soluble polymer.
  • the enzyme provided in the reaction layer 500 includes at least any one selected from among oxidoreductase, transferase, hydrolase, lyase, isomerase, and synthetase.
  • a buffer the pH of which ranges from 2 to 9, may be additionally used.
  • the reaction layer 500 includes an electron mediator, which reacts with the above enzyme and is thus oxidizable or reducible. The electron mediator is oxidized or reduced upon application of predetermined voltage to the working electrode, thus producing oxidation current or reduction current .
  • reaction layer according to the first embodiment of the present invention is applied equivalently to the following second to fourth embodiments .
  • FIG. 5 illustrates the cross-sections of the conductive coupling layer of the biosensor according to the present invention.
  • the conductive coupling layer 50 is used to couple the insulating substrate to the upper substrate so as to define the sample injection part and the sample reaction chamber. Further, the conductive coupling layer exposed to the reaction chamber is used as the counter electrode and the sample recognition electrode in the biosensor.
  • the conductive coupling layer 50 may be provided in the form of a single conductive layer 400 (FIG. 5 (a)), and exhibits both electrical conductivity and adhesiveness for coupling the insulating substrate and the upper substrate.
  • the conductive coupling layer 50 may be provided in the form of a multilayer structure.
  • a conductive coupling layer (FIG. 5 (b) ) in which a conductive layer 400 is formed on both surfaces of an insulating layer 450, or a conductive coupling layer (FIG. 5(c)) in which on one surface of an insulating layer 500 is formed a conductive layer 400 and on the other surface of the insulating layer is formed a non-conductive adhesion layer 500.
  • the above two layers 400, 500 are adhesive to the upper substrate or insulating substrate.
  • the conductive coupling layer may be used in the form in which the insulating layer 450 formed between the non-conductive adhesion layer 500 and the conductive layer 400 is removed (FIG. 5 (d) ) .
  • one side surface of the conductive coupling layer may be irregularly formed, in order to increase the contact area with the sample.
  • the conductive coupling layer preferably has a thickness ranging from 1 p to 1000 /zm. If the thickness of the conductive coupling layer is greater than 1000 ⁇ m, the area of the counter electrode in contact with the sample is increased, thus attaining stable electrical signals, but the volume of the formed reaction chamber is large, undesirably requiring a large amount of sample. Conversely, if the thickness is less than 1 ⁇ , the conductive coupling layer is too thin, and thus the force of adhesion between the insulating substrate 10 and the upper substrate 80 is low, undesirably making it difficult to couple the substrates. Moreover, attributable to a large error in the measuring of the thickness, the volume of the resultant reaction chamber is not uniform, and also, the area of the counter electrode coming into with the sample is reduced, and the conductive coupling layer has difficulty functioning as the counter electrode.
  • the sheet resistance of the conductive coupling layer preferably ranges from 10 ⁇ to 500 k ⁇ . If the sheet resistance of the conductive coupling layer is lower than 10 ⁇ , the ratio of the conductive material in the conductive coupling layer should be increased, and thereby the amount of adhesive material becomes proportionately decreased, undesirably resulting in low adhesiveness. Conversely, if the sheet resistance is greater than 500 k ⁇ , it is difficult to stably measure electrical signals attributable to the high resistance.
  • the adhesiveness of the conductive coupling layer 50 to the insulating substrate or the upper substrate is realized by applying heat, pressure, or UV to the conductive coupling layer.
  • the conductive layer 400 may be prepared in a combination of a conductive material such as a conductive polymer, gold, silver, silver chloride, palladium graphite, carbon, ITO particles, metal particles, or CNTs with an adhesive material, and may further include another available material.
  • FIG. 6 sequentially shows the process of manufacturing a biosensor including a sample recognition electrode for sensing the supply of a sample into a reaction chamber, according to the present invention.
  • a conductive material 30 for use as a working electrode in an electrochemical sensor On the surface of an insulating substrate 10, a conductive material 30 for use as a working electrode in an electrochemical sensor, a conductive material 20 which is electrically connected to conductive coupling layers 5OA, 5OB which are to be formed in a subsequent process to thus use the conductive coupling layers as a counter electrode, and a conductive material 110 which is electrically connected to a conductive coupling layer 5OE which is to be formed in a subsequent process to thus use the conductive coupling layer 5OE as a sample recognition electrode for sensing the supply of a sample into a sample reaction chamber 70 are formed in a pattern as shown in FIG. 6 (a) .
  • an insulating layer 40 is formed on portions of the conductive materials 20, 30, 110, thus forming a connector 2OA for electrical connection to conductive coupling layers 5OA, 5OB acting as a counter electrode, a connector IIOA for electrical connection to a conductive coupling layer 50E acting as a sample recognition electrode, and a working electrode 3OA.
  • the conductive coupling layers 5OA, 5OB, 5OE are formed in a pattern as shown in FIG. 6(c) .
  • the conductive coupling layers 5OA, 5OB are used as the counter electrode, and the conductive coupling layer 5OE is used as the sample recognition electrode for recognizing the supply of the sample.
  • the insulating substrate 10 is coupled to an upper substrate 80 by means of the conductive coupling layers 5OA, 5OB, 5OE disposed therebetween, thereby forming a sample reaction chamber
  • connection lines 2OB, 3OB, HOB for electrically connecting the conductive materials 20,
  • a material that reacts with the analyte is provided in the sample reaction chamber 70.
  • a reagent reacting with the analyte of the sample may be provided on the working electrode 3OA exposed to the sample reaction chamber 70 or the exposed upper substrate 80.
  • the sample is injected via the sample injection part 90 and is then supplied into the sample reaction chamber 70.
  • air in the reaction chamber is discharged to the outside through the air vent 60.
  • the complete supply of the sample is electrochemically sensed by the conductive coupling layer 5OE formed in the portion of the reaction chamber 70 to recognize the sample.
  • the sample is allowed to react in the reaction chamber for a predetermined period of time, and is then measured through an electrochemical method using the working electrode 3OA and the conductive coupling layers 5OA, 5OB for the counter electrode.
  • the sample recognition electrode is formed on the insulating substrate 10 or the upper substrate 80, a surplus area is utilized, ultimately increasing the volume of the sample reaction chamber 70.
  • the sample should be used in a relatively large amount.
  • the conductive coupling layer 5OE for recognizing the sample is formed to measure the analyte using a relatively small amount of the sample, there is no need to separately form the sample recognition electrode on the insulating substrate 10 or the upper substrate 80.
  • sample recognition electrode according to the first embodiment is applied equivalently to the following second to fourth embodiments.
  • FIG. 7 sequentially shows the process of manufacturing the biosensor including a porous layer which is adhered over the working electrode by the conductive coupling layer, according to the present invention .
  • conductive materials 20, 30 are formed in a pattern as shown in FIG. 7 (a) .
  • an insulating layer 40 is formed in a pattern as illustrated in FIG. 7 (b) on portions of the conductive materials, thus forming a working electrode 3OA, a connector 2OA for electrical connection to a conductive coupling layer 50 acting as a counter electrode, and connection lines 2OB, 3OB for electrically connecting the conductive materials to an electrochemical measurement system.
  • the conductive coupling layer 50 is formed on the insulating layer 40. As such, the conductive coupling layer 50 should be applied so that the working electrode 3OA is not covered therewith.
  • a porous layer 200 is adhered over the working electrode 3OA by the conductive coupling layer 50, after which an upper substrate is coupled, thus manufacturing the biosensor 100.
  • the porous layer 200 is microporous, and plays a role in removing fine particles present in the sample.
  • the porous layer is preferably formed of micropores having a size less than 10 ⁇ m to remove erythrocytes from the sample.
  • the porous layer preferably includes an enzyme for reacting with the analyte .
  • a serum component in which erythrocytes present in the whole blood sample are removed, is passed through the porous layer 200 to flow toward the working electrode 3OA, and also, the portion of the separated serum sample is supplied into the conductive coupling layer 50 acting as the counter electrode, thus electrically connecting the working electrode 3OA with the conductive coupling layer 50 acting as the counter electrode.
  • FIG. 8 sequentially shows the process of manufacturing the biosensor provided with a sample injection part and a sample reaction chamber at both sides thereof, according to the present invention.
  • conductive materials 20, 30 are formed in a pattern as shown in FIG. 8 (a) on an insulating substrate 10.
  • an insulating layer 40 is formed in a pattern as shown in FIG. 8 (b) on the conductive materials 20, 30, thus forming a working electrode 3OA, connectors 2OA for electrical connection to conductive coupling layers 5OA, 5OB acting as a counter electrode, and respective connection lines 2OB, 3OB for electrical connection of the conductive materials 20, 30 to a measurement system.
  • FIG. 9 sequentially shows a process of manufacturing a biosensor including a plurality of reaction chambers according to a second embodiment of the present invention .
  • a plurality of conductive materials 20, 30, 110 is formed in a pattern as shown in FIG. 9 (a) on an insulating substrate.
  • an insulating layer 40 is formed in a pattern as shown in FIG. 9(b) on the plurality of conductive materials, thus forming a plurality of working electrodes
  • 3OA a plurality of connectors 2OA, 3OA, IIOA for electrically connecting the plurality of conductive materials to a plurality of conductive coupling layers
  • FIG. 9 (c) the conductive coupling layers 5OA, 5OB, 5OE are formed on the insulating layer 40.
  • the insulating substrate 10 is coupled to an upper substrate 80 by means of the conductive coupling layers disposed therebetween, and thereby, the conductive coupling layer 5OE is formed as a sample recognition electrode for sensing the supply of the sample into the reaction chamber 70, and the conductive coupling layers 5OA, 5OB are formed as a counter electrode. Further, a plurality of air vents is formed to discharge air from the reaction chamber into which the sample is injected.
  • FIG. 10 sequentially shows a process of manufacturing a biosensor having a facing structure, according to a third embodiment of the present invention.
  • conductive materials 20, 30, 120 are formed in a pattern as shown in FIG. 10 (a) on an insulating substrate 10, and a conductive material 130 is formed in a pattern as shown in FIG. 10 (b) on an upper substrate
  • an insulating layer 40 is formed in a pattern as shown in FIG. 10 (c) on portions of the conductive materials 20, 30, 120, which are applied on the insulating substrate 10, thus forming a working electrode 3OA, connectors 2OA, 120A for electrical connection of the conductive materials 20, 120 to conductive coupling layers 5OA, 5OB, and connection lines 2OB, 3OB, 120B for electrical connection of the conductive materials 20, 30, 120 to a measurement system.
  • the conductive coupling layers 5OA, 5OB, 5OE are formed on the insulating layer 40 and the connectors 2OA, 120A, and the conductive coupling layer 5OE is not brought into direct contact with an analytic sample and is electrically connected with a connector 130B formed on the upper substrate 80.
  • an insulating layer 40 is formed in a pattern as shown in FIG. 10 (d) on the upper substrate 80, thus forming a working electrode 130A and a connector 130B for making an electrical connection to the conductive coupling layer 5OE.
  • the conductive coupling layers 5OA, 5OB, 5OE are formed in a pattern as shown in FIG. 10 (e) on the insulating substrate 10.
  • the insulating substrate 10 is coupled to the upper substrate 80 by means of the conductive coupling layers disposed therebetween, thereby manufacturing a biosensor including a sample injection part 90 for sample injection, a sample reaction chamber 70, and an air vent 60.
  • FIG. 11 shows the cross-section of the biosensor having a facing structure, according to the third embodiment of the present invention.
  • the insulating substrate 10 and the upper substrate 80 are coupled to each other by means of the conductive coupling layers 5OB, 5OA disposed therebetween, thereby defining the sample reaction chamber 70.
  • the conductive materials 130A, 3OA exposed to the reaction chamber 70 are used as the working electrode in the electrochemical sensor, and the portions 50C, 50D of the conductive coupling layers 5OA, 5OB, which are exposed toward the reaction chamber, are used as the counter electrode.
  • a plurality of working electrodes for analysis of the sample may be provided in the limited reaction chamber 70. Accordingly, this biosensor is advantageous because a plurality of analytes can be measured using a small amount of sample, and the portions 5OC, 5OD of the conductive coupling layers exposed to the reaction chamber are used as the counter electrode, thus obviating a need to manufacture the counter electrodes corresponding to the respective working electrodes. Furthermore, a reactive material that reacts with the analyte of the sample to generate electrically detectable signals may be provided on the working electrodes 3OA, 130A in the reaction chamber 70.
  • FIG. 12 sequentially shows a process of manufacturing a biosensor including a plurality of reaction chambers, according to a fourth embodiment of the present invention
  • FIG. 13 shows the cross- section of the biosensor including a plurality of reaction chambers, according to the fourth embodiment.
  • a plurality of conductive materials 20, 30 is formed in a pattern as shown in FIG. 12 (a) on an insulating substrate 10.
  • an insulating layer 40 is formed in a pattern as illustrated in FIG. 12 (b) on the plurality of conductive materials 20, 30, thus forming a plurality of working electrodes 3OA, connectors 2OA to a conductive coupling layer 50, and a plurality of connection lines 2OB, 3OB for electrical connection of the conductive materials 20, 30 to a measurement system.
  • the conductive coupling layer 50 is formed on the insulating layer 40.
  • the insulating substrate 10 is coupled to an upper substrate 80 by means of the conductive coupling layer disposed therebetween, thereby forming a plurality of sample reaction chambers 70.
  • portions 5OF (FIG. 13) of the conductive coupling layer 50, which are exposed to the plurality of reaction chambers 70, are used as a counter electrode.
  • This biosensor is advantageous in that a plurality of samples can be measured at once .
  • FIG. 14 illustrates the biosensor according to the present invention, which is mounted in a rotatable rotator .
  • the biosensor according to the present invention includes the insulating substrate 10, the insulating layer 40, the conductive coupling layer
  • the conductive material for use as the working electrode may be formed on either or both of the insulating substrate and the upper substrate.
  • the insulating layer 40 is formed on the insulating substrate 10 or the upper substrate 80, thus defining the conductive material for a plurality of working electrodes and the plurality of conductive materials for electrical connection to the conductive coupling layer 50, and forming the plurality of connection lines 2OB, 3OB for electrical connection to a measurement system. Thereafter, a plurality of flow pathways 310, 310A for transferring the analytic sample or fluid, the plurality of sample reaction chambers, and the air vents 60 for discharge of air from the plurality of reaction chambers are formed using the conductive coupling layer 50.
  • the portion of the conductive coupling layer 50 which is exposed to the flow pathways 310, 310A and the reaction chambers 70, plays a role as the counter electrode in the electrochemical biosensor for measurement of the analyte in the sample, and the flow pathways 310, 310A, the reaction chambers 70, and the air vents 60 are defined by coupling the insulating substrate 10 to the upper substrate 80 by means of the conductive coupling layer 50 disposed therebetween.
  • transfer, mixing, reaction, and pretreatment of the sample may be realized by force caused by the rotation of the sensor, and further, the rotation of the sensor results in multiple electrical connections to the measurement system.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Electrochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Hematology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un biodétecteur, dans lequel une couche de raccordement permettant de réunir un substrat supérieur et un substrat inférieur, dont l'un ou l'autre est doté d'une électrode de travail, comprend un matériau conducteur et est donc utilisée en tant que contre-électrode ou en tant qu'électrode de reconnaissance d'un échantillon. Le biodétecteur comprend au moins une électrode de travail qui est constituée soit sur un substrat supérieur soit sur un substrat isolant soit sur les deux et qui est utilisée en vue de la mesure de signaux générés par un analyte ; et au moins une couche de raccordement conductrice, qui est utilisée pour réunir les deux substrats et qui est caractérisée par une conductivité électrique. Dans le biodétecteur, au moins une enceinte réactionnelle destinée à la réaction de l'analyte est délimitée par les deux substrats réunis et la couche de raccordement conductrice. L'une des surfaces latérales de la couche de raccordement conductrice faisant face à l'enceinte réactionnelle est utilisée en tant que contre-électrode en vue de la mesure des signaux générés par l'analyte.
PCT/KR2008/002862 2008-04-28 2008-05-22 Biodétecteur WO2009133983A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR20080039332A KR100998648B1 (ko) 2008-04-28 2008-04-28 바이오센서
KR10-2008-0039332 2008-04-28

Publications (1)

Publication Number Publication Date
WO2009133983A1 true WO2009133983A1 (fr) 2009-11-05

Family

ID=41255186

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2008/002862 WO2009133983A1 (fr) 2008-04-28 2008-05-22 Biodétecteur

Country Status (2)

Country Link
KR (1) KR100998648B1 (fr)
WO (1) WO2009133983A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012028841A1 (fr) * 2010-08-30 2012-03-08 Cilag Gmbh International Bandelette de détection d'analyte équipée d'une électrode à deux sections électriques distinctes
CN104034764A (zh) * 2014-06-13 2014-09-10 上海师范大学 一种具有靶向和可视化双功能的电化学细胞传感器及其制备方法
CN105466985A (zh) * 2014-08-28 2016-04-06 立威生技实业股份有限公司 用于生物检测试片的电极与其制造方法
CN106124596A (zh) * 2016-06-30 2016-11-16 英太格电子科技（苏州）有限公司 一种生化测试片的制备工艺

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014046318A1 (fr) 2012-09-21 2014-03-27 ㈜ 더바이오 Procédé de reconnaissance d'échantillon et biocapteur l'utilisant
KR101448243B1 (ko) 2012-09-21 2014-10-08 (주) 더바이오 시료를 인식하는 방법 및 이를 이용한 바이오센서
KR102136321B1 (ko) * 2013-08-19 2020-07-22 엘지전자 주식회사 전기화학적 면역분석 카트리지 및 이것의 제조방법
KR102130384B1 (ko) * 2018-04-24 2020-07-07 한국원자력연구원 바이오센서 전극의 제조방법, 바이오센서 전극, 및 이를 포함하는 바이오 센서
KR102247002B1 (ko) * 2019-10-15 2021-04-29 동우 화인켐 주식회사 바이오 센서

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0643131A (ja) * 1992-07-22 1994-02-18 Fujitsu Ltd バイオセンサ、バイオセンサ集合体、及び、その製造方法
US5846708A (en) * 1991-11-19 1998-12-08 Massachusetts Institiute Of Technology Optical and electrical methods and apparatus for molecule detection
US7063776B2 (en) * 2003-06-17 2006-06-20 Chun-Mu Huang Structure and manufacturing method of disposable electrochemical sensor strip
JP2007174990A (ja) * 2005-12-28 2007-07-12 Matsushita Electric Ind Co Ltd 細胞電気生理センサアレイおよびその製造方法
US7348183B2 (en) * 2000-10-16 2008-03-25 Board Of Trustees Of The University Of Arkansas Self-contained microelectrochemical bioassay platforms and methods

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5846708A (en) * 1991-11-19 1998-12-08 Massachusetts Institiute Of Technology Optical and electrical methods and apparatus for molecule detection
JPH0643131A (ja) * 1992-07-22 1994-02-18 Fujitsu Ltd バイオセンサ、バイオセンサ集合体、及び、その製造方法
US7348183B2 (en) * 2000-10-16 2008-03-25 Board Of Trustees Of The University Of Arkansas Self-contained microelectrochemical bioassay platforms and methods
US7063776B2 (en) * 2003-06-17 2006-06-20 Chun-Mu Huang Structure and manufacturing method of disposable electrochemical sensor strip
JP2007174990A (ja) * 2005-12-28 2007-07-12 Matsushita Electric Ind Co Ltd 細胞電気生理センサアレイおよびその製造方法

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012028841A1 (fr) * 2010-08-30 2012-03-08 Cilag Gmbh International Bandelette de détection d'analyte équipée d'une électrode à deux sections électriques distinctes
CN104034764A (zh) * 2014-06-13 2014-09-10 上海师范大学 一种具有靶向和可视化双功能的电化学细胞传感器及其制备方法
CN105466985A (zh) * 2014-08-28 2016-04-06 立威生技实业股份有限公司 用于生物检测试片的电极与其制造方法
CN106124596A (zh) * 2016-06-30 2016-11-16 英太格电子科技（苏州）有限公司 一种生化测试片的制备工艺

Also Published As

Publication number Publication date
KR20090113550A (ko) 2009-11-02
KR100998648B1 (ko) 2010-12-07

Similar Documents

Publication Publication Date Title
WO2009133983A1 (fr) Biodétecteur
US6863801B2 (en) Electrochemical cell
USRE42567E1 (en) Electrochemical cell
US8354012B2 (en) Electrochemical cell
US6174420B1 (en) Electrochemical cell
KR100340174B1 (ko) 전기화학적 바이오센서 테스트 스트립, 그 제조방법 및 전기화학적 바이오센서
AU2002367214B2 (en) Micro-band electrode
EP2414534B1 (fr) Dispositif de test électrochimique
AU738128B2 (en) Electrochemical cell

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08753655

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08753655

Country of ref document: EP

Kind code of ref document: A1