Biosensor for detecting glucose in body fluid, manufacturing method thereof, and system for measuring concentration of glucose in body fluid using same

Abstract

Provided is a biosensor for detecting glucose in body fluid by using a measuring device, the biosensor comprising: a substrate; a plurality of electrodes positioned on the substrate; and a reagent layer, wherein the reagent layer comprises: an oxidation-reduction enzyme; and an electron transfer medium including a ruthenium compound and neutral red.

Description

A biosensor for detecting glucose in body fluids, a method for manufacturing the same, and a system for measuring the concentration of glucose in body fluids using the same

[0001] One embodiment relates to a biosensor for detecting glucose in bodily fluids and a method for manufacturing the same. Another embodiment relates to a system for measuring the concentration of glucose in bodily fluids using the biosensor.

[0002] Quantitatively or qualitatively analyzing analytes present in body fluids is a crucial task both chemically and clinically. Examples include measuring blood sugar levels in diabetic patients and cholesterol, a risk factor for various adult diseases.

[0003] In electrochemical biosensors utilizing enzyme activity (e.g., glucose sensor, uric acid sensor, protein sensor, DNA sensor, sucrose sensor, glutamate-oxaloacetate transaminase (GOT) or glutamate-pyruvate transaminase (GPT) sensor, etc.), it is very important to measure enzyme activity more quickly and reproducibly by reacting with a sample (e.g., a specific substance present in a body fluid including saliva or blood).

[0004] Accordingly, the biosensor includes a reagent layer for reacting with a sample to cause a chemical or physical change and an electrode structure for measuring the change as an electric current.

[0005] The principle of sample measurement of these biosensors is to estimate the concentration of a sample (e.g., glucose) in a body fluid by measuring the current generated in response to the sample reacting with the reagent layer. Therefore, current sensing technology is required to improve the accuracy of sample measurement.

[0006] One embodiment comprises a reagent layer comprising an electron transfer mediator including an oxidoreductase, a ruthenium compound, and neutral red to implement current sensing for improving the accuracy of sample measurement.

[0007] At this time, one embodiment proposes a reagent layer that controls the concentration of an electron transfer mediator so as to satisfy the conditions that the increase in current corresponding to an increase in the concentration of a sample in a body fluid must be greater than or equal to a detectable increase rate and that the current must be greater than or equal to a detectable threshold value for a reference concentration of the sample in the body fluid.

[0008] However, the technical problems to be solved by the present invention are not limited to the above problems, and can be expanded in various ways without departing from the technical spirit and scope of the present invention.

[0009] According to one embodiment, the reagent layer provided in the electrode structure of the biosensor may include an electron transfer mediator including an oxidoreductase; a ruthenium compound, and neutral red.

[0010] According to one aspect, the concentration of the neutral red may be adjusted so that the rate at which the current generated in response to the redox reaction of the sample in the body fluid with the redox enzyme increases in response to the increase in the concentration of the sample is greater than a preset rate of increase.

[0011] According to another aspect, the concentration of the neutral red may be adjusted so that the current generated in response to the redox reaction of the sample with the redox enzyme is above a preset threshold value.

[0012] According to another aspect, the ruthenium compound may be characterized by including hexaammineruthenium trichloride.

[0013] According to another aspect, it may be characterized in that FAD-GDH (Flavin Adenine Dinucleotide-dependent Glucose Dehydrogenase) is used as the above oxidoreductase.

[0014] According to one embodiment, a biosensor may be characterized by comprising: a substrate; an electrode structure provided on the substrate, including a working electrode and a reference electrode; and a reagent layer provided on the electrode structure, wherein the reagent layer includes an oxidoreductase; and an electron transfer mediator including a ruthenium compound and neutral red.

[0015] According to one embodiment, a method for manufacturing a biosensor comprises the steps of: providing an electrode structure including a working electrode and a reference electrode on a substrate; and providing a reagent layer on the electrode structure, wherein the step of providing the reagent layer on the electrode structure is characterized by providing the reagent layer with an electron transfer mediator including an oxidoreductase, a ruthenium compound, and neutral red.

[0016] According to one aspect, the step of providing the reagent layer may be characterized by including a step of adjusting the concentration of the neutral red so that the rate at which the current generated in response to the redox reaction of the sample in the body fluid with the redox enzyme increases in response to the increase in the concentration of the sample is greater than a preset increase rate.

[0017] According to another aspect, the step of providing the reagent layer may be characterized by including a step of adjusting the concentration of the neutral red so that the current generated in response to the redox reaction of the sample with the redox enzyme is greater than a preset threshold value.

[0018] According to one embodiment, a biosensor for detecting glucose in a body fluid using a measuring device is provided, comprising: a substrate; a plurality of electrodes positioned on the substrate; and a reagent layer; wherein the reagent layer includes: an oxidoreductase; and an electron transfer mediator including a ruthenium compound and neutral red.

[0019] According to one side, the concentration of the neutral red may be 0.1 mM or more and 1 mM or less.

[0020] According to another aspect, the electrode is for transmitting a current signal generated by a redox reaction between the glucose and the reagent layer to the measuring device, and the concentration of the glucose may be determined based on the value of the current signal transmitted to the measuring device.

[0021] According to another aspect, the value of the current signal may change in proportion to the concentration of the glucose within a numerical range of 357 nA to 3601 nA when the concentration of the glucose changes within a numerical range of 40 mg / dL to 600 mg / dL.

[0022] According to another aspect, the increase rate of the current signal according to the concentration of the glucose may be 3 nA / (mg / dL) or more and 6 nA / (mg / dL) or less.

[0023] According to another aspect, the ruthenium compound may be at least one compound selected from the group consisting of Hexaammineruthenium trichloride, Ruthenium(III) Hexaamine, Ruthenium(II) Tris(bipyridine), Ruthenium(II) Tris(4,4'-dicarboxy-2,2'-bipyridine), and Ruthenium(III) Chloro Di(2,2'-bipyridine).

[0024] According to another aspect, the oxidoreductase may be glucose dehydrogenase (GDH).

[0025] According to another aspect, the oxidoreductase may be at least one compound selected from the group including FAD-GDH (Flavin Adenine Dinucleotide-dependent Glucose Dehydrogenase), NAD-GDH (Nicotinamide Adenine Dinucleotide-dependent Glucose Dehydrogenase), and PQQ-GDH (Pyrroloquinoline Quinone-dependent Glucose Dehydrogenase).

[0026] According to one embodiment, a system for measuring the concentration of glucose in a body fluid is provided, comprising: a body fluid collection device; a biosensor of any one of the biosensors described above; and a measuring device.

[0027] According to one embodiment, a method for manufacturing a biosensor for detecting glucose in a body fluid is provided, comprising: preparing a substrate; forming a plurality of electrodes on the substrate; and arranging a reagent layer on the electrodes, wherein the reagent layer comprises: an oxidoreductase; and an electron transfer mediator including a ruthenium compound and neutral red.

[0028] According to one side, the concentration of the neutral red may be 0.1 mM or more and 1 mM or less.

[0029] One embodiment can implement current sensing for improved accuracy of sample measurement by forming a reagent layer with an electron transfer mediator including an oxidoreductase and a ruthenium compound and neutral red.

[0030] At this time, one embodiment may propose a reagent layer that adjusts the concentration of an electron transfer mediator so as to satisfy the condition that the increase in current corresponding to the increase in the concentration of the sample in the body fluid must be greater than or equal to a detectable increase rate and the condition that the current must be greater than or equal to a detectable threshold value for the reference concentration of the sample in the body fluid.

[0031] However, the effects of the present invention are not limited to the above effects, and can be expanded in various ways without departing from the technical spirit and scope of the present invention.

[0032] FIG. 1 is a diagram illustrating an example of a biosensor system according to one embodiment.

[0033] FIG. 2 is a block diagram illustrating an example of a biosensor included in a biosensor system according to one embodiment.

[0034] FIG. 3 is a diagram illustrating the structure of neutral red used as an electron transfer medium in a reagent layer included in a biosensor according to one embodiment.

[0035] Figures 4 and 5 are drawings for explaining the electron transfer excellence of neutral red used as an electron transfer medium in a reagent layer included in a biosensor according to one embodiment.

[0036] FIG. 6 is a drawing for explaining how the concentration of neutral red used as an electron transfer medium in a reagent layer included in a biosensor according to one embodiment is controlled.

[0037] Figure 7 is a flow chart illustrating a method for manufacturing a biosensor according to one embodiment.

[0038] Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings. However, the present invention is not limited or restricted by these embodiments. In addition, the same reference numerals in each drawing represent the same components.

[0039] In addition, the terminology used in this specification is a term used to appropriately express embodiments of the present invention, and this may vary depending on the intention of the viewer or operator, or the customs of the field to which the present invention belongs. Therefore, the definition of these terms should be determined based on the contents throughout this specification. For example, in this specification, the singular also includes the plural unless specifically stated in the phrase. In addition, the terms "comprises" and / or "comprising" as used herein do not exclude the presence or addition of one or more other components, steps, operations, and / or elements to the mentioned components, steps, operations, and / or elements. In addition, although the terms first, second, etc. are used in this specification to describe various regions, directions, shapes, etc., these regions, directions, and shapes should not be limited by these terms. These terms are only used to distinguish a certain region, direction, or shape from another region, direction, or shape. Therefore, a part referred to as a first part in one embodiment may be referred to as a second part in another embodiment.

[0040] It should also be understood that the various embodiments of the present invention, while different, are not necessarily mutually exclusive. For example, specific shapes, structures, and characteristics described herein may be implemented in other embodiments without departing from the spirit and scope of the present invention. Furthermore, it should be understood that the location, arrangement, or configuration of individual components within each of the disclosed embodiments may be modified without departing from the spirit and scope of the present invention.

[0041] Hereinafter, the body fluid may include saliva or blood. Specifically, the body fluid may include various body substances such as blood, lymph, plasma, serum, saliva, urine, feces, sweat, mucus, tears, cerebrospinal fluid, nasal secretions, cervical or vaginal secretions, semen, pleural effusion, amniotic fluid, ascites, middle ear fluid, joint fluid, and gastric contents extracts, but is not limited thereto. In addition, the sample below may include glucose. Specifically, the sample below is a target substance to be detected through a biosensor, and may include various physiological indicators and substances used to measure environmental conditions such as glucose, lactate, alcohol, hemoglobin, cardiolipin, protein, insulin, estrogen, and other hormones, microorganisms, DNA / RNA, calcium, sodium, pH, oxygen, and carbon dioxide, but is not limited thereto. Furthermore, the term "saliva sugar" hereinafter refers to glucose contained in saliva, and "blood sugar" refers to glucose contained in blood. Furthermore, "acquiring saliva sugar" hereinafter refers to obtaining information about saliva sugar (e.g., saliva sugar concentration, index, value, etc.), and "predicting blood sugar" refers to predicting information about blood sugar (e.g., blood sugar concentration, index, value, etc.).

[0042] FIG. 1 is a diagram illustrating an example of a biosensor system according to one embodiment. The biosensor system of FIG. 1 represents an example including a plurality of electronic devices (110, 120, 130, 140, 150), a server (160), and a network (170). FIG. 1 is an example for explaining the invention, and the number of electronic devices or servers is not limited to that of FIG. 1.

[0043] A plurality of electronic devices (110, 120, 130, 140, 150) are terminals carried by each of the test subjects, and each of the plurality of electronic devices (110, 120, 130, 140, 150) may include a body fluid collection device or collection device (111), a biosensor (112), and a measurement device (113) as a specimen measurement system.

[0044] The collection device (111) is a device for collecting bodily fluids including saliva or blood. It may be a collection tool including a specimen collection unit, a filter, and a compression tube, or a lancing device including a lancet. If the specimen is saliva, the collection device (111) may remove interfering substances from the collected saliva through a filter in the compression tube, and provide the saliva from which the interfering substances have been removed to the outside, i.e., to a biosensor (112). If the specimen is blood, the collection device (111) may make a hole of a predetermined depth in the skin, such as the edge of the fingertip of the test subject, through a lancet, and provide the blood collected on the skin to the biosensor (112).

[0045] The biosensor (112) is a component that performs the function of sensing a sample to be measured (e.g., glucose) when a body fluid including saliva or blood is injected into the biosensor (112) through a collection device (111).

[0046] When a biosensor (112) is inserted into the measuring device (113), the measuring device (113) recognizes this and switches to a standby state to recognize the injection of body fluid by providing a specific signal to the biosensor (112). When body fluid is injected into the biosensor (112), the measuring device (113) recognizes that the body fluid has been injected through a response signal (e.g., change in resistance or voltage) generated from the biosensor (112). When the measuring device (113) determines that body fluid has been injected, it applies a voltage to the working electrode and the reference electrode for measuring a sample (e.g., glucose). As a result, a response signal (e.g., response current) is generated from the biosensor (112) according to the redox reaction between the sample and the reagent layer in the body fluid, and the measuring device (113) receives this to detect the sample (e.g., glucose) or measure or predict the concentration of the sample.

[0047] For this purpose, the measuring device (113) may include a memory, a processor, an input / output interface, etc.

[0048] And, the measuring device (113) may include a communication module to communicate with the server (160) via the network (170). However, instead of directly connecting to the network (170) and communicating with the server (160), a plurality of electronic devices (110, 120, 130, 140, 150) each including the measuring device (113) may be configured to communicate with a terminal (e.g., a smart phone, a mobile phone, a tablet PC, a navigation device, a computer, a laptop, a digital broadcasting terminal, a PDA (Personal Digital Assistant), a PMP (Portable Multimedia Player), etc.) possessed by the subject via the network (170), or may be configured to connect to the network (170) via the terminal and communicate with the server (160).

[0049] The communication method is not limited, and may include not only a communication method that utilizes a communication network (e.g., a mobile communication network, a wired Internet, a wireless Internet, and a broadcasting network) that the network (170) may include, but also short-range wireless communication between devices. For example, the network (170) may include any one or more of a network such as a Personal Area Network (PAN), a Local Area Network (LAN), a Campus Area Network (CAN), a Metropolitan Area Network (MAN), a Wide Area Network (WAN), a Broadband Network (BBN), and the Internet. In addition, the network (170) may include any one or more of a network topology including, but not limited to, a bus network, a star network, a ring network, a mesh network, a star-bus network, a tree, or a hierarchical network.

[0050] Although each of the above multiple electronic devices (110, 120, 130, 140, 150) has been described as including a collection device (111), a biosensor (112), and a measuring device (113), the present invention is not limited thereto, and the measuring device (113) may include a terminal (e.g., a smart phone, a mobile phone, a tablet PC, a navigation device, a computer, a laptop, a digital broadcasting terminal, a PDA (Personal Digital Assistant), a PMP (Portable Multimedia Player), etc.) carried by the test subject.

[0051] The server (160) may be implemented as a computer device or a plurality of computer devices that communicate with a plurality of electronic devices (110, 120, 130, 140, 150) through a network (170) to receive measurement information of a sample in a body fluid measured from a measurement device (113) of each of the plurality of electronic devices (110, 120, 130, 140, 150), and provide a biomarker measurement service that predicts a biomarker (e.g., blood sugar) of a subject based on the received measurement information.

[0052] That is, each of the plurality of electronic devices (110, 120, 130, 140, 150) can access the server (160) under the control of a pre-installed operating system (OS) or at least one program (e.g., firmware, a browser or application installed under the OS) and receive a biomarker measurement service provided by the server (160).

[0053] FIG. 2 is a block diagram showing an example of a biosensor included in a biosensor system according to one embodiment, FIG. 3 is a diagram showing the structure of neutral red used as an electron transfer mediator of a reagent layer included in a biosensor according to one embodiment, FIGS. 4 and 5 are diagrams for explaining the electron transfer excellence of neutral red used as an electron transfer mediator of a reagent layer included in a biosensor according to one embodiment, and FIG. 6 is a diagram for explaining that the concentration of neutral red used as an electron transfer mediator of a reagent layer included in a biosensor according to one embodiment is controlled.

[0054] Referring to FIG. 2, the biosensor (200) may include a substrate (210), an electrode structure (220), and a reagent layer (230).

[0055] The substrate (210) is a base on which an electrode structure (220) is provided, and can be formed of an insulating material so that a working electrode and a reference electrode included in the electrode structure (220) are electrically separated from each other.

[0056] If the working electrode and the reference electrode are formed independently on planes of different heights instead of being formed on the same plane, the substrate (210) may be composed of a plurality of layers so that the working electrode and the reference electrode are arranged.

[0057] The electrode structure (220) may be configured to promote a redox reaction between a sample and a reagent layer (230) in a body fluid through interaction with the sample and reagent layer (230) in the body fluid and to transmit a current signal generated by the redox reaction to a measuring device.

[0058] That is, the electrode structure (220) is a component provided on the substrate (210) including a working electrode and a reference electrode, through which the current generated by the reagent layer (230) provided on the working electrode and the reference electrode reacting with a specimen in a body fluid can be measured.

[0059] Accordingly, the arrangement structure and shape of the working electrode and the reference electrode can be implemented in various ways as long as they satisfy the condition that enables measurement of the current generated by the reagent layer (230) provided on the working electrode and the reference electrode reacting with the specimen in the body fluid.

[0060] The reagent layer (230) is provided on the electrode structure (220) and may include substances that cause a redox reaction with a sample in a body fluid injected onto the reagent layer (230). More specifically, the reagent layer (230) may include an oxidoreductase and an electron transfer mediator.

[0061] The oxidoreductase may include glucose dehydrogenase (GDH). Specifically, the oxidoreductase may include at least one compound selected from the group consisting of FAD-GDH (Flavin Adenine Dinucleotide-dependent Glucose Dehydrogenase), NAD-GDH (Nicotinamide Adenine Dinucleotide-dependent Glucose Dehydrogenase), and PQQ-GDH (Pyrroloquinoline Quinone-dependent Glucose Dehydrogenase). However, the present invention is not limited thereto, and the oxidoreductase may include various enzymes that cause a redox reaction with a sample in a body fluid.

[0062] The electron transport mediator may include a ruthenium compound. Specifically, the ruthenium compound may include at least one compound selected from the group consisting of Hexaammineruthenium trichloride, Ruthenium(III) Hexaamine, Ruthenium(II) Tris(bipyridine), Ruthenium(II) Tris(4,4'-dicarboxy-2,2'-bipyridine), and Ruthenium(III) Chloro Di(2,2'-bipyridine). However, the present invention is not limited thereto, and the electron transfer medium may include various compounds that mediate the rapid transfer of electrons generated by the reaction of the reagent layer (230) containing the redox enzyme and the specimen in the body fluid injected onto the reagent layer (230) to the electrode structure (220).

[0063] The electron transfer mediator may further include neutral red. Neutral red is 3-Amino-7-dimethylamino-2-methylphenazine hydrochloride (C) as shown in FIG. 3 15 H 17 It can have the structure of ClN4).

[0064] The graphs shown in Figures 4 to 6 are based on the data described in [Table 1] to [Table 3], respectively.

[0065]

[0066]

[0067]

[0068] In this way, the biosensor (200) according to one embodiment uses neutral red as an electron transfer medium in the reagent layer (230), and unlike the case where neutral red is not used as an electron transfer medium as shown in FIG. 4, based on the characteristic that the sensing current rapidly increases in response to the concentration of the sample, the condition that the increase in current corresponding to the increase in the concentration of the sample in the body fluid must be greater than the detectable increase rate is satisfied, thereby improving the accuracy of sample measurement.

[0069] In addition, a biosensor (200) according to one embodiment uses neutral red as an electron transfer mediator instead of using an electron transfer mediator such as DCIP (2,6-Dichloro-4-[(4-hydroxyphenyl)imino]cyclohexa-2,5-dien-1-one), Toluidine blue (7-amino-8-methylphenothiazin-3-ylidene)-dimethylammonium chloride), NBT (2,2'-bis(4-Nitrophenyl)-5,5'-diphenyl-3,3'-(3,3'-dimethoxy-4,4'-diphenylene)ditetrazolium chloride), and AQDS (Disodium Anthraquinone-2,6-disulfonate), and thus can have a characteristic of rapidly increasing the sensing current in response to the concentration of the sample, as shown in FIG. 5, compared to the case where an electron transfer mediator such as DCIP, Toluidine blue, NBT, and AQDS is used. Accordingly, the biosensor (200) can improve the accuracy of sample measurement by satisfying the condition that the increase in current corresponding to the increase in the concentration of the sample in the body fluid must be greater than the detectable increase rate based on the characteristic that the sensing current rapidly increases in response to the concentration of the sample.

[0070] To this end, the concentration of neutral red used as an electron transfer medium can be adjusted so that the rate at which the current generated in response to the redox reaction of the sample with the oxidoreductase increases in response to the increase in the concentration of the sample in the body fluid is greater than a preset rate of increase.

[0071] More specifically, the current generated in response to the redox reaction of the sample with the oxidoreductase increases in proportion to the increase in the concentration of the sample in the body fluid and also increases in proportion to the increase in the concentration of neutral red. For example, the current generated in response to the redox reaction of the sample with the oxidoreductase increases in proportion to the increase in the concentration of the sample in the body fluid and also increases in proportion to the increase in the concentration of neutral red, as shown in Fig. 6. Specifically, the current generated in response to the redox reaction of the sample with the oxidoreductase has a larger value when the concentration of neutral red is 1 mM than when it is 0.1 mM.

[0072] Accordingly, the rate at which the current generated in response to the redox reaction of the sample with the oxidoreductase increases in response to the increase in the concentration of the sample in the body fluid increases in proportion to the increase in the concentration of neutral red. That is, the rate of increase in current is greater when the concentration of neutral red is 1 mM than when it is 0.1 mM.

[0073] A larger current increase rate like this means ease of current sensing, and the concentration of neutral red can be adjusted so that the current generated in response to the redox reaction of the sample with the oxidoreductase increases at a rate greater than or equal to a preset increase rate (a critical increase rate that allows accurate current sensing) in response to an increase in the concentration of the sample in the body fluid. For example, the concentration of neutral red can be adjusted to 0.1 mM at which the current increase rate is greater than or equal to the preset increase rate (a critical increase rate that allows accurate current sensing). More preferably, the concentration of neutral red can be adjusted to 1 mM at which the current increase rate is greater than or equal to the preset increase rate (a critical increase rate that allows accurate current sensing).

[0074] However, the concentration of neutral red is not limited or restricted thereto, and may be adjusted so that the current generated in response to the redox reaction of the sample with the oxidoreductase is higher than a preset threshold value (minimum current value for current sensing) for the reference concentration of the sample in the body fluid (concentration within the average range including the sample in the body fluid). This corresponds to the minimum condition for the biosensor (200) to detect the sensing of the target substance in the sample. For example, the concentration of neutral red may be adjusted to be 0.1 mM or more and 1 mM or less.

[0075] As illustrated in FIG. 6 and described in [Table 3], when the reagent layer contains 0.1 mM or more and 1 mM or less of neutral red, the value of the current signal generated in response to the redox reaction of the sample with the oxidoreductase may vary in proportion to the concentration of glucose within a numerical range of 357 nA or more and 3601 nA or less when the concentration of glucose varies within a numerical range of 40 mg / dL or more and 600 mg / dL or less. At this time, the increase rate of the current signal according to the concentration of glucose may be 3 nA / (mg / dL) or more and 6 nA / (mg / dL) or less.

[0076] Figure 7 is a flow chart illustrating a method for manufacturing a biosensor according to one embodiment.

[0077] The manufacturing method described below refers to a process for manufacturing a biosensor described with reference to FIGS. 1 to 6, and is assumed to be performed by an automated and mechanized manufacturing system.

[0078] Referring to FIG. 7, in the first step (S710), the manufacturing system can provide an electrode structure including a working electrode and a reference electrode to a substrate. Providing an electrode structure including a working electrode and a reference electrode to a substrate means placing and forming an electrode structure including a working electrode and a reference electrode on the substrate.

[0079] In the second step (S720), the manufacturing system can provide a reagent layer to the electrode structure. Providing a reagent layer to the electrode structure means arranging and forming a reagent layer on the electrode structure.

[0080] More specifically, in the second step (S720), the manufacturing system can provide a reagent layer comprising a ruthenium compound and neutral red as an oxidoreductase and an electron transfer mediator.

[0081] The oxidoreductase may include glucose dehydrogenase (GDH). Specifically, the oxidoreductase may include at least one compound selected from the group consisting of FAD-GDH (Flavin Adenine Dinucleotide-dependent Glucose Dehydrogenase), NAD-GDH (Nicotinamide Adenine Dinucleotide-dependent Glucose Dehydrogenase), and PQQ-GDH (Pyrroloquinoline Quinone-dependent Glucose Dehydrogenase). However, the present invention is not limited thereto, and the oxidoreductase may include various enzymes that cause a redox reaction with a sample in a body fluid.

[0082] The electron transport mediator may include a ruthenium compound. Specifically, the ruthenium compound may be at least one compound selected from the group consisting of Hexaammineruthenium trichloride, Ruthenium(III) Hexaamine, Ruthenium(II) Tris(bipyridine), Ruthenium(II) Tris(4,4'-dicarboxy-2,2'-bipyridine), and Ruthenium(III) Chloro Di(2,2'-bipyridine). However, the electron transfer mediator is not limited thereto, and may include various compounds that mediate the rapid transfer of electrons generated by the reaction between the sample in the body fluid injected onto the reagent layer and the reagent layer containing the redox enzyme to the electrode structure.

[0083] The electron transfer mediator may further include neutral red. Neutral red is 3-Amino-7-dimethylamino-2-methylphenazine hydrochloride (C) as shown in FIG. 3 15 H 17 It can have the structure of ClN4).

[0084] In particular, the manufacturing system can enable accurate current sensing by controlling the concentration of neutral red so that the rate at which the current generated in response to the redox reaction of the sample with the redox enzyme in the second step (S720) increases in response to the increase in the concentration of the sample in the body fluid is greater than a preset rate of increase.

[0085] Additionally, without being limited or restricted thereto, the manufacturing system may enable current sensing for a sample having a reference concentration by adjusting the concentration of neutral red so that the current generated in response to the sample undergoing a redox reaction with an oxidoreductase in the second step (S720) is greater than a preset threshold value for the reference concentration of the sample in the body fluid.

[0086] The device including the measuring device (113) described above may be implemented as hardware components, software components, and / or a combination of hardware components and software components. For example, the devices and components described in the embodiments may be implemented using one or more general-purpose computers or special-purpose computers, such as a processor, a controller, an Arithmetic Logic Unit (ALU), a Digital Signal Processor, a microcomputer, a Field Programmable Array (FPA), a Programmable Logic Unit (PLU), a microprocessor, or any other device capable of executing and responding to instructions. The processing device may execute an operating system (OS) and one or more software applications running on the OS. In addition, the processing device may access, store, manipulate, process, and generate data in response to the execution of the software. For ease of understanding, the processing device is sometimes described as being used alone, but one of ordinary skill in the art will recognize that the processing device may include multiple processing elements and / or multiple types of processing elements. For example, a processing unit may include multiple processors, or a processor and a controller. Other processing configurations, such as parallel processors, are also possible.

[0087] Software may include a computer program, code, instructions, or a combination of one or more of these, and may configure a processing device to perform a desired operation or, independently or collectively, command a processing device. The software and / or data may be embodied in any type of machine, component, physical device, virtual equipment, computer storage medium, or device to be interpreted by the processing device or to provide instructions or data to the processing device. The software may be distributed over networked computer systems and stored or executed in a distributed manner. The software and data may be stored on one or more computer-readable recording media.

[0088] The method according to the embodiment may be implemented in the form of program commands that can be executed through various computer means and recorded on a computer-readable medium. The computer-readable medium may include program commands, data files, and data structures, either singly or in combination. The program commands recorded on the medium may be those specially designed and configured for the embodiment or may be those known and available to those skilled in the art of computer software. Examples of the computer-readable recording medium include magnetic media such as hard disks, floppy disks, and magnetic tapes, optical media such as CD-ROMs and DVDs, magneto-optical media such as floptical disks, and hardware devices specially configured to store and execute program commands such as ROMs, RAMs, and flash memories. Examples of program commands include not only machine language codes generated by a compiler, but also high-level language codes that can be executed by a computer using an interpreter, etc.

[0089] Although the embodiments described above have been described by way of limited examples and drawings, those skilled in the art will appreciate that various modifications and variations can be made based on the above description. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, or if the components of the described systems, structures, devices, and circuits are combined or combined in a different manner than described, or if they are replaced or substituted with other components or equivalents.

[0090] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

1. A biosensor for detecting glucose in body fluid using a measuring device, substrate; a plurality of electrodes positioned on the substrate; and reagent layer; Including, The above reagent layer is, Oxidoreductase; and Electron transfer mediators including ruthenium compounds and neutral red; Including, A biosensor for detecting glucose in body fluids using a measuring device.

2. In paragraph 1, The concentration of the above neutral red is 0.1 mM or more and 1 mM or less. A biosensor for detecting glucose in body fluids using a measuring device.

3. In paragraph 1, The above electrode is for transmitting a current signal generated by a redox reaction between the glucose and the reagent layer to the measuring device, The concentration of the above glucose is determined based on the value of the current signal transmitted to the measuring device. A biosensor for detecting glucose in body fluids using a measuring device.

4. In paragraph 3, The value of the above current signal changes in proportion to the concentration of the glucose within a numerical range of 357 nA to 3601 nA when the concentration of the glucose changes within a numerical range of 40 mg / dL to 600 mg / dL. A biosensor for detecting glucose in body fluids using a measuring device.

5. In paragraph 3, The increase rate of the current signal according to the concentration of the glucose is 3 nA / (mg / dL) or more and 6 nA / (mg / dL) or less. A biosensor for detecting glucose in body fluids using a measuring device.

6. In paragraph 1, The above ruthenium compound is at least one compound selected from the group consisting of Hexaammineruthenium trichloride, Ruthenium(III) Hexaamine, Ruthenium(II) Tris(bipyridine), Ruthenium(II) Tris(4,4'-dicarboxy-2,2'-bipyridine) and Ruthenium(III) Chloro Di(2,2'-bipyridine). A biosensor for detecting glucose in body fluids using a measuring device.

7. In paragraph 1, The above oxidoreductase is glucose dehydrogenase (GDH). A biosensor for detecting glucose in body fluids using a measuring device.

8. In paragraph 7, The above oxidoreductase is at least one compound selected from the group consisting of FAD-GDH (Flavin Adenine Dinucleotide-dependent Glucose Dehydrogenase), NAD-GDH (Nicotinamide Adenine Dinucleotide-dependent Glucose Dehydrogenase), and PQQ-GDH (Pyrroloquinoline Quinone-dependent Glucose Dehydrogenase). A biosensor for detecting glucose in body fluids using a measuring device.

9. In a system for measuring the concentration of glucose in body fluid, fluid collection device; A biosensor as described in any one of claims 1 to 8; and measuring device; Including, A system for measuring the concentration of glucose in body fluids.

10. A method for manufacturing a biosensor for detecting glucose in body fluid, Steps to prepare the substrate; a step of forming a plurality of electrodes on the substrate; and A step of placing a reagent layer on the above electrode; Including, The above reagent layer is, Oxidoreductase; and Electron transfer mediators including ruthenium compounds and neutral red; Including, A method for manufacturing a biosensor for detecting glucose in body fluid.

11. In Article 10, The concentration of the above neutral red is 0.1 mM or more and 1 mM or less. A method for manufacturing a biosensor for detecting glucose in body fluid.

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