KR100998648B1 - biosensor - Google Patents
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- KR100998648B1 KR100998648B1 KR20080039332A KR20080039332A KR100998648B1 KR 100998648 B1 KR100998648 B1 KR 100998648B1 KR 20080039332 A KR20080039332 A KR 20080039332A KR 20080039332 A KR20080039332 A KR 20080039332A KR 100998648 B1 KR100998648 B1 KR 100998648B1
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- 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
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- 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
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Abstract
The present invention relates to a biosensor in which a conductive material is formed on a bonding layer for bonding an upper substrate or a lower substrate provided with a working electrode and used as an auxiliary electrode or a sample recognition electrode.
At least one working electrode formed on at least one of an upper substrate and an insulating substrate, and configured to measure a signal formed by the analyte; At least one conductive bonding layer coupling the two substrates and having electrical conductivity; The combined two substrates and the conductive bonding layer form a reaction space of at least one analyte, and one side of the conductive bonding layer exposed to the reaction space assists in measuring a signal formed by the analyte. Characterized in that formed into an electrode.
Biosensor, Conductive Bonding Layer
Description
The present invention relates to a biosensor, and more particularly, to a biosensor used as an auxiliary electrode or a sample recognition electrode by forming a conductive material on a bonding layer for bonding an upper substrate or a lower substrate having a working electrode.
Background arts related to biosensors include: The following numbers are all US registered patent numbers.
5,437,999; After the electrode is patterned on the insulator substrate using a photolithography method with a precise area of a working eletrode or a counter electrode, the substrate on which the electrodes are formed is bonded to each other using an intermediate bonding layer. To form an electrode. On one side of the intermediate bonding layer is formed an empty space for injection of the measurement sample, the empty space is formed with a reagent reacting with the analyte, the air discharge port for the air discharge on the substrate on which the working electrode or auxiliary electrode is formed Formed.
5,582,697; A biosensor for quantifying an enzyme substrate present in a sample by an electrochemical method, a working electrode, an auxiliary electrode, and a third electrode on an insulating substrate, and a reaction layer including an oxidoreductase Biosensor formed on the working electrode and the auxiliary electrode. The third electrode is used as an electrode for sensing the sample injection, and is located at least far from the working electrode and the auxiliary electrode from the sample inlet.
6,071,391 and 6,156,173; The upper substrate and the lower substrate are joined by an intermediate bonding layer to form a space for sample injection, and a working electrode or auxiliary electrode is formed on the two substrates to face each other, wherein the two substrates have air The discharge port is not formed, and the substrate provided with the working electrode and the auxiliary electrode at the portion injecting the sample has a sharp tip. In addition, in order to connect the electrode formed on the upper substrate to the conductive wire formed on the lower substrate, a hole is formed in a portion of the bonding layer to connect the conductive material.
6,576,101; In the biosensor measuring the sample electrochemically using a sample of 1uL or less, a method of manufacturing the electrode in the form of a coating like an enzyme participating in the reaction on the working electrode by binding a redox material that can be oxidized in the air to the polymer In addition, the oxidation / reduction material bound to the polymer serves as an electron transfer material (mediator) when reacted with a working electrode or an enzyme, and has a property of not diffusing into a sample solution.
6,618,934; In a method of manufacturing a plurality of electrochemical sensors, a plurality of working electrodes and auxiliary electrodes are formed on the substrate in the first electrode region and the second electrode region, a spacer layer is formed in one of the electrode regions, and then the sample After removing a portion of the spacer to form a reaction chamber, and folding the substrate to form a layer of the first electrode zone and the second electrode zone to produce an electrochemical sensor, each sensor is separated into It consists of at least one working electrode, auxiliary electrode and sample reaction chamber.
6,863,800; In a biosensor configuration in which an analyte is positioned between an electrode formed on an electrode substrate and an electrode formed on a substrate to cover the electrode, the working electrode includes an ink composed of a reagent, a conductive material, and an electron mediator reacting with the analyte. The reference electrode is formed by coating a material reacting with an analyte and an electron transporting material on a conductive material, and the auxiliary electrode is positioned to face an electrode substrate made of a conductive material and to cover each other.
6,885,196; In the biosensor structure in which the first insulating substrate on which the working electrode is formed and the second insulating substrate on which the auxiliary electrode is formed face each other, a sample inflow space is formed between the two substrates, and the working electrode and the auxiliary electrode are formed in the sample inflow space. The reaction layer including the electrode and the oxidoredutase is exposed, and the distance between the working electrode and the auxiliary electrode is 150 micrometers or less, and the area of the auxiliary electrode exposed to the sample inflow space is larger than that of the exposed working electrode. It is a small biosensor.
6,942,769B2; In the biosensor assembled by combining an electrode, an insulating layer, an adhesive layer, a layer having a porosity of 10 to 40%, an adhesive layer, and a cover plate in this order after forming an electrode on the substrate for electrode formation, the insulating film Holes are formed in the cover plate and the porous layer controls the flow and volume of the analyte by holes formed in the insulating film and the cover plate.
7,022,218; In the biosensor for the analysis of the trace amount, the working electrode having a plurality of branches and the auxiliary electrode having a plurality of branches are alternately arranged alternately on the first insulating substrate, and another auxiliary electrode branch is formed on the second insulating substrate. The two substrates are assembled to face each other, and a working electrode and an auxiliary electrode are alternately arranged with a reaction reagent composed of a oxidoreductase in a sample inflow space formed between the two substrates.
7,050,843; An insulating first substrate having a conductive surface and a second substrate having a conductive surface are bonded to each other by an intermediate layer, and the capillary channel formed by the two substrates and the intermediate layer has a biosensor in which a reagent reacting with a sample is dried. And forming an insulating pattern line having a “V” shape on the conductive surface to adjust the flow of the sample, and dividing the conductive surface into two sections.
2006/0008581 A1; In the method of manufacturing an electrochemical sensor, having a working electrode on the first insulator and forming a plurality of insulating layers on the working electrode, the hole is formed through the working electrode to form a container, the cross section of the cut working electrode The container is exposed to the wall of the container, and additionally, a reference electrode is formed on the uppermost layer of the plurality of insulators, and the bottom plate is additionally bonded.
The biosensor measuring the analytical material electrochemically described in the prior art, basically consists of an upper insulator substrate, a lower insulator substrate and an intermediate layer that combines the two substrates, the sample injection or sample by the two substrates and the intermediate layer Comprising a space for the reaction, the two substrates are provided with a working electrode and an auxiliary electrode for measuring the electrochemical oxidation or reduction.
In particular, in order to measure the electrochemical oxidation / reduction current, the area where the working electrode and the auxiliary electrode formed on the substrate are in contact with the sample is important, and in general, the electrochemical signal increases in proportion to the electrode area. In order to obtain a larger electrochemical signal using the same volume of the sample, the patents include a working electrode on one of the two substrates, an auxiliary electrode on the other substrate, and then use the intermediate layer to connect the two electrodes to each other. Form facing each other. The arrangement of the electrodes has an advantage of maximizing the electrode area more efficiently than forming the two electrodes on the same substrate.
However, since the conductive material used in most of the electrodes is opaque, when the electrode is formed on both substrates, there is a disadvantage that it is difficult to visually check when the sample is injected.
To solve this problem, a method of forming an electrode other than the working electrode and the auxiliary electrode to confirm the sample injection is used, but the area of the working electrode or the auxiliary electrode is reduced or measured due to the formation of a third electrode in a limited area. There is a disadvantage that the amount of sample for the increase.
In order to solve this problem, an object of the present invention is to provide a biosensor to form a conductive material in the bonding layer for coupling the upper substrate or lower substrate provided with a working electrode to use as an auxiliary electrode or a sample recognition electrode. .
Biosensor according to the present invention for solving the above problems,
In the biosensor for measuring the analyte in the sample,
At least one working electrode formed on at least one of an upper substrate and an insulating substrate, and configured to measure a signal formed by the analyte; At least one conductive bonding layer combining the two substrates and having electrical conductivity; The combined two substrates and the conductive bonding layer form a reaction space of at least one analyte, and one side of the conductive bonding layer exposed to the reaction space assists in measuring a signal formed by the analyte. Characterized in that formed into an electrode.
In addition, the working electrode is characterized in that it comprises at least any one of a conductive polymer, gold, palladium, graphite, carbon, ITO particles, metal particles, carbon nanotubes (carbon nanotube, CNT).
In addition, irregularities or bends are formed on one side of the conductive bonding layer to increase the contact area with the sample.
In addition, the conductive bonding layer is formed by printing or coating a conductive adhesive material on both sides of the insulating layer.
In addition, the conductive bonding layer is characterized in that the conductive adhesive material is formed on one surface of the insulating layer, the non-conductive adhesive material is formed on the other surface of the insulating layer.
In addition, the conductive bonding layer is characterized in that formed only with a conductive adhesive material.
In addition, the conductive bonding layer is characterized in that it comprises at least one of a conductive polymer, gold, silver, silver chloride, palladium, graphite, carbon, ITO particles, metal particles, carbon nanotubes (carbon nanotube, CNT).
In addition, the coupling between the upper substrate and the insulating substrate, and the conductive bonding layer is characterized in that the coupling using pressure, heat, or light.
In addition, the reaction space is characterized in that a reaction layer comprising at least one enzyme reacting with the analyte, and the electron transfer material capable of oxidation or reduction by reacting with the enzyme. In this case, the reaction layer is preferably formed under the reaction space and the conductive bonding layer. In this case, the reaction layer preferably contains a water-soluble polymer. In this case, the enzyme contained in the reaction layer is preferably at least one enzyme of oxidoreductase, transfer enzyme, hydrolase, degrading enzyme, isomerase, synthetase. In this case, the reaction layer preferably further includes a buffer material of
In addition, the thickness of the conductive bonding layer is characterized in that 1μm to 1000μm.
In addition, the sheet resistance of the conductive bonding layer is characterized in that 10Ω to 500kΩ.
In addition, it is formed on the working electrode, it characterized in that it further comprises a porous layer bonded by the conductive bonding layer. At this time, the porous layer is preferably made of microporous of less than 10 ㎛ capable of removing red blood cells in the sample. In this case, the porous layer preferably includes an enzyme that reacts with the analyte in the sample.
In addition, the upper substrate, the insulating substrate, and the conductive bonding layer formed in the reaction space formed by the conductive coupling layer, characterized in that it further comprises a sample recognition electrode for detecting a sample introduced into the reaction space; do. In this case, the sample recognition electrode is formed spaced apart from the sample injection unit for the sample injection, it is preferable to be formed at a position farther than the working electrode.
The apparatus may further include an air outlet connected to the upper substrate, the insulating substrate, and the reaction space formed by the conductive bonding layer.
According to the biosensor according to the present invention as described above, since the conductive bonding layer is used as an auxiliary electrode or a sample recognition electrode, there is no need to separately provide the electrodes on the substrate, thereby maximizing the area of the working electrode.
Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. First, it should be noted that the same components or parts in the drawings represent the same reference numerals as much as possible. In describing the present invention, detailed descriptions of related well-known functions or configurations are omitted in order not to obscure the gist of the present invention.
1 is a process diagram illustrating a process of manufacturing a biosensor according to a first embodiment of the present invention, FIG. 2 is a cross-sectional view of a biosensor according to a first embodiment of the present invention, and FIG. 3 is a first embodiment of the present invention. 4 is a cross-sectional view illustrating that a reaction layer is formed in a reaction space in a biosensor according to an example, and FIG. 4 illustrates that a reaction layer is formed below a portion of a reaction space and a conductive coupling layer in a biosensor according to a first embodiment of the present invention. 5 is a cross-sectional view showing a conductive bonding layer of the biosensor according to the present invention, FIG. 6 is a biosensor including a sample recognition electrode for detecting a sample introduced into the reaction space in the biosensor according to the present invention. 7 is a process diagram illustrating a process for manufacturing a biosensor formed on a working electrode in the biosensor according to the present invention and including a porous layer bonded by a conductive bonding layer. 8 is a process diagram illustrating a process of manufacturing a biosensor having a sample injection unit and a sample reaction space on both sides of the biosensor in the biosensor according to the present invention, and FIG. Process diagram showing a process of manufacturing a biosensor having a plurality of reaction spaces according to a second embodiment, Figure 10 is a process diagram showing a process of manufacturing a face-type biosensor according to a third embodiment of the present invention 11 is a cross-sectional view of a facing biosensor according to a third exemplary embodiment of the present invention, and FIG. 12 is a process diagram illustrating a process of manufacturing a biosensor having a plurality of reaction spaces according to a fourth exemplary embodiment of the present invention. 13 is a cross-sectional view of a biosensor having a plurality of reaction spaces according to a fourth embodiment of the present invention, and FIG. 14 is a diagram illustrating a biosensor according to the present invention mounted on a rotatable rotor.
1 is a process diagram illustrating a process of manufacturing a biosensor according to a first embodiment of the present invention.
Referring to FIG. 1A, first, the
Next, an insulating
Then, the conductive bonding material is formed in a pattern as shown in FIG. 1C to form the
The conductive bonding layer is a layer made of a material having electrical conductivity and adhesion, which will be described later with reference to FIG. 5.
Next, as shown in FIG. 1 (d), the insulating
2 is a cross-sectional view of a biosensor according to a first embodiment of the present invention.
Referring to FIG. 2, the
3 is a cross-sectional view showing a reaction layer is formed in the reaction space in the biosensor according to the first embodiment of the present invention, Figure 4 is a portion of the reaction space and the conductive bonding layer in the biosensor according to the first embodiment of the present invention It is sectional drawing which shows that the reaction layer was formed in the lower part.
3 and 4, a
In particular, as shown in FIG. 4, when
At this time, the enzyme provided in the
The description of the reaction layer is equally applicable to the second to fourth embodiments described below as well as the first embodiment of the present invention.
5 is a cross-sectional view showing a conductive bonding layer of the biosensor according to the present invention.
Referring to FIG. 5, the
In addition, the
In addition, irregularities or bends may be formed on one side of the conductive bonding layer to increase the contact area with the sample.
In addition, the thickness of the conductive bonding layer is preferably 1μm to 1000μm. Because, when the thickness of the conductive bonding layer is more than 1000μm, the area of the auxiliary electrode in contact with the sample is increased to obtain a stable electrical signal, but the volume of the formed reaction space is required a large amount of samples, 1μm or less Due to the low thickness of the bonding layer, it is difficult to bond due to low adhesion between the insulating
In addition, the sheet resistance of the conductive bonding layer is preferably 10Ω to 500kΩ. If the surface resistance of the conductive bonding layer is less than 10Ω, the ratio of the conductive material to the conductive bonding layer should be increased. As a result, the content of the adhesive material decreases, so that the adhesive strength decreases, and if the resistance is more than 500 kΩ, the electrical resistance is high and stable. This is because signal measurement is difficult.
Adhesiveness of the insulating substrate or the upper substrate of the
The description of the conductive bonding layer may be equally applied to the second to fourth embodiments described below as well as the first embodiment.
6 is a process diagram illustrating a process of manufacturing a biosensor including a sample recognition electrode for sensing a sample introduced into the reaction space in the biosensor according to the present invention.
First, the
Next, as shown in FIG. 6B, an insulating
Then, the conductive bonding layers 50A, 50B, and 50E are formed in a pattern as shown in FIG. 6 (c). The
Then, as shown in Figure 6 (d), by coupling the insulating
In addition, a material reacting with the analyte to measure the analyte of the sample may be configured in the
The sample is injected through the
When the electrode for sample recognition is formed on the insulating
In order to solve this problem and form a
In addition, the description of the sample recognition electrode can be equally applied to the second to fourth embodiments described below as well as the first embodiment of the present invention.
7 is a process diagram illustrating a process of manufacturing a biosensor formed on a working electrode in the biosensor according to the present invention and including a porous layer bonded by a conductive bonding layer.
First,
Next, the insulating
Next, as shown in FIG. 7C, the
Then, as shown in Figure 7 (d), after bonding the
For example, when a whole blood sample is applied onto the
8 is a process diagram illustrating a process of manufacturing a biosensor having a sample injection unit and a sample reaction space on both sides of the biosensor according to the present invention.
First, the
Next, the insulating
Next, as illustrated in FIG. 8C,
Then, by using the conductive bonding layer to combine the insulating
9 is a flowchart illustrating a process of manufacturing a biosensor having a plurality of reaction spaces according to a second embodiment of the present invention.
First, a plurality of
Next, an insulating
Next, as shown in FIG. 9C, conductive bonding layers 50A, 50B, and 50E are formed on the insulating
Then, the
10 is a flowchart illustrating a process of manufacturing a face-to-face biosensor according to a third embodiment of the present invention.
First,
Next, an insulating
Next, an insulating
Next, conductive bonding layers 50A, 50B, and 50E are formed on the insulating
Next, as shown in FIG. 10 (f), the insulating
FIG. 11 is a cross-sectional view of the large-area biosensor according to the third embodiment. Referring to this, the insulating
In the biosensor having such a structure, since a plurality of sample electrodes may be provided in a
12 is a flowchart illustrating a process of manufacturing a biosensor having a plurality of reaction spaces according to a fourth embodiment of the present invention, and FIG. 13 is provided with a plurality of reaction spaces according to the fourth embodiment of the present invention. A cross section of the biosensor.
First, a plurality of
Next, an insulating
Next, as shown in FIG. 12 (c), a
Then, the plurality of
In the
14 is a view showing a biosensor according to the present invention mounted on a rotatable rotor.
Referring to FIG. 14, the biosensor according to the present invention includes an insulating
The biosensor forms an insulating
In addition, the exposed portions of the
The
As described above with reference to the drawings illustrating a biosensor according to the present invention, the present invention is not limited by the embodiments and drawings disclosed herein, it is various within the technical scope of the present invention by those skilled in the art Of course, modifications can be made.
1 is a process chart showing a process of manufacturing a biosensor according to a first embodiment of the present invention;
2 is a cross-sectional view of a biosensor according to a first embodiment of the present invention;
3 is a cross-sectional view showing that the reaction layer is formed in the reaction space in the biosensor according to the first embodiment of the present invention;
4 is a cross-sectional view illustrating that a reaction layer is formed below a portion of a reaction space and a conductive coupling layer in a biosensor according to a first embodiment of the present invention;
5 is a cross-sectional view showing a conductive bonding layer of a biosensor according to the present invention;
6 is a process diagram illustrating a process of manufacturing a biosensor including a sample recognition electrode for sensing a sample introduced into the reaction space in the biosensor according to the present invention;
7 is a process diagram illustrating a process of manufacturing a biosensor formed on a working electrode in the biosensor according to the present invention and including a porous layer bonded by a conductive bonding layer;
8 is a process chart showing a process of manufacturing a biosensor having a sample injection unit and a sample reaction space on both sides of the biosensor in the biosensor according to the present invention;
9 is a process diagram illustrating a process of manufacturing a biosensor having a plurality of reaction spaces according to a second embodiment of the present invention;
10 is a process chart showing a process of manufacturing a face-to-face biosensor according to a third embodiment of the present invention;
11 is a cross-sectional view of a face type biosensor according to a third embodiment of the present invention;
12 is a flowchart illustrating a process of manufacturing a biosensor having a plurality of reaction spaces according to a fourth embodiment of the present invention;
13 is a cross-sectional view of a biosensor having a plurality of reaction spaces according to a fourth embodiment of the present invention;
14 is a view showing a biosensor according to the present invention mounted on a rotatable rotor.
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KR20080039332A KR100998648B1 (en) | 2008-04-28 | 2008-04-28 | biosensor |
PCT/KR2008/002862 WO2009133983A1 (en) | 2008-04-28 | 2008-05-22 | Biosensor |
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Cited By (3)
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KR101448243B1 (en) | 2012-09-21 | 2014-10-08 | (주) 더바이오 | Method to recognze sample and bio sensor using the same |
US9395321B2 (en) | 2012-09-21 | 2016-07-19 | The Bio Co. Ltd. | Sample recognition method and biosensor using same |
WO2021075944A1 (en) * | 2019-10-15 | 2021-04-22 | 동우 화인켐 주식회사 | Biosensor |
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US20120048746A1 (en) * | 2010-08-30 | 2012-03-01 | Cilag Gmbh International | Analyte test strip with electrically distinguishable divided electrode |
KR102136321B1 (en) * | 2013-08-19 | 2020-07-22 | 엘지전자 주식회사 | Electrochemical immunoassay cartridge and manufactruring method thereof |
CN104034764B (en) * | 2014-06-13 | 2016-03-30 | 上海师范大学 | One has target and visual bifunctional electro-chemical cells sensor and preparation method thereof |
TW201608237A (en) * | 2014-08-28 | 2016-03-01 | 立威生技實業股份有限公司 | Electrode for biosensor and method for manufacturing the same |
CN106124596A (en) * | 2016-06-30 | 2016-11-16 | 英太格电子科技(苏州)有限公司 | A kind of preparation technology of biochemical test sheet |
KR102130384B1 (en) * | 2018-04-24 | 2020-07-07 | 한국원자력연구원 | Method for manufacturing biosensor electrode, biosensor electrode, and biosensor comprising thereof |
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US5846708A (en) * | 1991-11-19 | 1998-12-08 | Massachusetts Institiute Of Technology | Optical and electrical methods and apparatus for molecule detection |
US7348183B2 (en) * | 2000-10-16 | 2008-03-25 | Board Of Trustees Of The University Of Arkansas | Self-contained microelectrochemical bioassay platforms and methods |
JP4821319B2 (en) * | 2005-12-28 | 2011-11-24 | パナソニック株式会社 | Cell electrophysiological sensor array and manufacturing method thereof |
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US7063776B2 (en) * | 2003-06-17 | 2006-06-20 | Chun-Mu Huang | Structure and manufacturing method of disposable electrochemical sensor strip |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
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KR101448243B1 (en) | 2012-09-21 | 2014-10-08 | (주) 더바이오 | Method to recognze sample and bio sensor using the same |
US9395321B2 (en) | 2012-09-21 | 2016-07-19 | The Bio Co. Ltd. | Sample recognition method and biosensor using same |
WO2021075944A1 (en) * | 2019-10-15 | 2021-04-22 | 동우 화인켐 주식회사 | Biosensor |
KR20210044569A (en) * | 2019-10-15 | 2021-04-23 | 동우 화인켐 주식회사 | Bio sensor |
KR102247002B1 (en) | 2019-10-15 | 2021-04-29 | 동우 화인켐 주식회사 | Bio sensor |
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