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

Abstract

Provided is a biosensor for detecting glucose in a body fluid using a measuring device, which includes: a substrate; a plurality of electrodes on the substrate; and a reagent layer; the reagent layer includes: an oxidoreductase; and an electron transfer mediator containing a ruthenium compound and Neutral red.

Description

Technical Field

[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 glucose concentration in bodily fluids using the aforementioned biosensor. Background Technology

[0002] Quantitative or qualitative analysis of substances present in body fluids is important both chemically and clinically. Examples include measuring blood glucose levels in diabetic patients and measuring cholesterol, a contributing factor to many adult diseases.

[0003] In electrochemical biosensors that utilize enzyme activity (e.g., glucose sensors, uric acid sensors, protein sensors, DNA sensors, sucrose sensors, glutamate-Oxaloacetate transaminase (GOT) or glutamate-Pyruvate transaminase (GPT) sensors), it is crucial to measure enzyme activity more rapidly and reproducibly by reacting with a detection medium (e.g., a specific substance present in bodily fluids containing saliva or blood).

[0004] Therefore, a biosensor includes a reagent layer for reacting with a detection substance to cause chemical or physical changes, and an electrode structure for measuring such changes in an electric current manner.

[0005] The principle of this biosensor for measuring the detector is to calculate the concentration of the detector (e.g., glucose) in the body fluid by using the current generated in response to the reaction between the detector and the reagent layer. Therefore, current sensing technology is needed to improve the accuracy of detector measurement. Summary of the Invention

[0006] Technical issues

[0007] To achieve current sensing for improving the accuracy of detector measurements, one embodiment uses an oxidoreductase and an electron transport medium containing ruthenium compounds and neutral red to form a reagent layer.

[0008] At this point, one embodiment proposes a reagent layer that adjusts the concentration of the electron transport medium to satisfy the conditions that "the increase in current corresponding to the increase in the concentration of the detector in the body fluid must be above a perceptible rate of increase" and "the current relative to the reference concentration of the detector in the body fluid must be above a perceptible threshold".

[0009] However, the technical problem to be solved by the present invention is not limited to the above-mentioned problem, and various extensions can be made without departing from the technical concept and scope of the present invention.

[0010] Technical solutions

[0011] According to one embodiment, the reagent layer provided to the biosensor electrode structure may include: an oxidoreductase; and an electron transport medium comprising a ruthenium compound and neutral red.

[0012] According to one aspect, the concentration of the neutral red may be characterized in that it is adjusted such that the current generated in response to the redox reaction between the detector in the body fluid and the redox enzyme increases at a rate greater than a predetermined rate of increase corresponding to an increase in the concentration of the detector.

[0013] According to another aspect, the concentration of the neutral red may be characterized in that it is adjusted such that the current generated in response to the redox reaction between the detector and the oxidoreductase reaches or exceeds a predetermined threshold.

[0014] According to another aspect, the ruthenium compound may be characterized by containing hexaammineruthenium trichloride.

[0015] According to another aspect, the oxidoreductase may be characterized by the use of FAD-GDH (flavin adenine dinucleotide-dependent glucose dehydrogenase).

[0016] According to one embodiment, a biosensor may be characterized by comprising: a substrate; an electrode structure comprising a working electrode and a reference electrode disposed on the substrate; and a reagent layer disposed on the electrode structure, the reagent layer comprising: an oxidoreductase; and an electron transport medium comprising a ruthenium compound and neutral red.

[0017] According to one embodiment, a method for manufacturing a biosensor may be characterized by including: providing an electrode structure comprising a working electrode and a reference electrode to a substrate; and providing a reagent layer to the electrode structure, wherein the step of providing the reagent layer to the electrode structure is to provide the reagent layer using an oxidoreductase and an electron transport medium comprising a ruthenium compound and neutral red.

[0018] According to one aspect, the step of providing the reagent layer may be characterized by including: adjusting the concentration of the neutral red such that the current generated in response to the redox reaction between the detector in the body fluid and the redox enzyme increases at a rate corresponding to an increase in the concentration of the detector, reaching or exceeding a predetermined increase rate.

[0019] According to another aspect, the step of providing the reagent layer may be characterized by including: adjusting the concentration of the neutral red such that the current generated in response to the redox reaction between the detector and the oxidoreductase reaches or exceeds a predetermined threshold.

[0020] According to one embodiment, a biosensor for detecting glucose in body fluids using a measuring device is provided, comprising: a substrate; a plurality of electrodes located on the substrate; and a reagent layer; the reagent layer comprising: an oxidoreductase; and an electron transport medium containing a ruthenium compound and neutral red.

[0021] According to one aspect, the concentration of the neutral red can be above 0.1 mM and below 1 mM.

[0022] According to another aspect, the electrode is used to transmit a current signal generated by the redox reaction between the glucose and the reagent layer to the measuring device, wherein the concentration of the glucose is determined based on the value of the current signal transmitted to the measuring device.

[0023] According to another aspect, when the concentration of glucose varies within a range of 40 mg / dL to 600 mg / dL, the value of the current signal can vary proportionally to the concentration of glucose within a range of 357 nA to 3601 nA.

[0024] According to another aspect, the rate of increase of the current signal based on the glucose concentration can be 3 nA / (mg / dL) or more and 6 nA / (mg / dL) or less.

[0025] According to another aspect, the ruthenium compound may be selected from at least one compound from the group consisting of Hexaammineruthenium trichloride, Ruthenium(III) Hexaamine, Ruthenium(II) Tris(bipyridine) (2,2'-bipyridine), Ruthenium(II) Tris(4,4'-dicarboxy-2,2'-bipyridine) (4,4'-dicarboxy-2,2'-bipyridine) (Ruthenium(III) Chloro Di(2,2'-bipyridine) ( ...

[0026] According to another aspect, the oxidoreductase may be glucose dehydrogenase (GDH).

[0027] According to another aspect, the oxidoreductase may be 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).

[0028] According to one embodiment, a system for measuring glucose concentration in body fluids is provided, comprising: a body fluid collection device; any one of the above-mentioned biosensors; and a measuring device.

[0029] According to one embodiment, a method for manufacturing a biosensor for detecting glucose in body fluids is provided, comprising: a step of preparing a substrate; a step of forming a plurality of electrodes on the substrate; and a step of disposing a reagent layer on the electrodes; wherein the reagent layer comprises: an oxidoreductase; and an electron transport medium comprising a ruthenium compound and neutral red.

[0030] According to one aspect, the concentration of the neutral red can be above 0.1 mM and below 1 mM.

[0031] Beneficial effects of the invention

[0032] One embodiment enables current sensing for improving the accuracy of detector measurements by constructing a reagent layer using an oxidoreductase and an electron transport medium containing a ruthenium compound and neutral red.

[0033] At this point, one embodiment can propose a reagent layer that adjusts the concentration of the electron transport medium to satisfy the conditions that "the increase in current corresponding to the increase in the concentration of the detector in the body fluid must be above a perceptible rate of increase" and "the current relative to the reference concentration of the detector in the body fluid must be above a perceptible threshold".

[0034] However, the effects of the present invention are not limited to those described above, and various extensions can be made without departing from the technical concept and scope of the present invention. Attached Figure Description

[0035] Figure 1 This is a diagram illustrating an example of a biosensor system according to one embodiment.

[0036] Figure 2 This is a block diagram illustrating an example of a biosensor included in a biosensor system according to one embodiment.

[0037] Figure 3 This is a diagram illustrating the structure of neutral red, which serves as an electron transport medium, in a reagent layer included in a biosensor according to one embodiment.

[0038] Figures 4 to 5 This is a diagram illustrating the superior electron transport properties of neutral red, which serves as an electron transport medium, in a reagent layer included in a biosensor according to an embodiment.

[0039] Figure 6 This is a diagram illustrating the adjustment of the concentration of neutral red, which serves as an electron transport medium, in a reagent layer included in a biosensor according to one embodiment.

[0040] Figure 7 This is a flowchart illustrating a method for manufacturing a biosensor according to one embodiment. Detailed Implementation

[0041] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the present invention is not limited or restricted by the embodiments. Furthermore, the same reference numerals in the various drawings denote the same parts.

[0042] Furthermore, the terminology used in this specification is for the purpose of appropriately expressing embodiments of the invention and may vary depending on the intention of the observer, operator, or convention in the field to which this invention pertains. Therefore, the definitions of these terms should be based on the entire text of this specification. For example, in this specification, unless specifically mentioned in the text, the singular form also includes the plural form. Furthermore, the use of "comprising" and / or "including" in this specification does not exclude the presence or addition of one or more other constituent elements, steps, actions, and / or components besides those stated. Additionally, although terms such as "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 merely used to distinguish a specific region, direction, or shape from other regions, directions, or shapes. Therefore, a portion referred to as a first part in one embodiment may also be referred to as a second part in another embodiment.

[0043] Furthermore, it should be understood that the various embodiments of the present invention are different from each other, but are not necessarily mutually exclusive. For example, specific shapes, structures, and characteristics described herein, when referring to one embodiment, can be implemented in other embodiments without departing from the spirit and scope of the present invention. Moreover, it should be understood that within the scope of the various embodiments presented, the position, arrangement, or configuration of individual constituent elements can be changed without departing from the spirit and scope of the present invention.

[0044] The following body fluids can include saliva or blood. Specifically, the following body fluids can include, but are not limited to, various substances in the body such as blood, lymph, plasma, serum, saliva, urine, feces, sweat, mucus, tears, cerebrospinal fluid, nasal secretions, cervical or vaginal secretions, semen, pleural fluid, amniotic fluid, ascites, middle ear fluid, synovial fluid, and gastric contents extract. In addition, the following detectors can include glucose. Specifically, the following detectors, as substances to be detected by biosensors, can include substances used to measure various physiological indicators and environmental conditions such as glucose, lactate, ethanol, hemoglobin, cardiolipin, protein, hormones such as insulin and estrogen, microorganisms, DNA / RNA, calcium, sodium, pH, oxygen, and carbon dioxide, but are not limited to these. Furthermore, the following terms refer to saliva glucose (as in saliva) and blood glucose (as in blood glucose). Additionally, the following terms refer to obtaining information about saliva glucose (e.g., concentration, index, value, etc.) and predicting blood glucose (e.g., concentration, index, value, etc.).

[0045] Figure 1 This is a diagram illustrating an example of a biosensor system according to one embodiment. Figure 1 The biosensor system shown is an example comprising multiple electronic devices (110, 120, 130, 140, 150), a server (160), and a network (170). Figure 1 For the purpose of illustrating the invention, the number of electronic devices or servers is not limited to one example. Figure 1 As shown.

[0046] Multiple electronic devices (110, 120, 130, 140, 150) serve as terminals held by the test subjects. Each of the multiple electronic devices (110, 120, 130, 140, 150) serves as a measurement system for the test subject, which may include a body fluid collection device or collection device (111), a biosensor (112), and a measurement device (113).

[0047] The collection device (111), as a device for collecting bodily fluids containing saliva or blood, can be a collection tool including a sample collection section, a filter, and a compression tube, or a lancing device including a lancet. When the sample is saliva, the collection device (111) can remove interfering substances from the collected saliva within the compression tube using the filter, and provide the saliva, now free of interfering substances, to the external biosensor (112). When the sample is blood, the collection device (111) can puncture the skin, such as the fingertip edge, of the subject to be tested using the lancet to a predetermined depth, and provide the blood clotted on the skin to the biosensor (112).

[0048] The biosensor (112) is a component that performs the function of sensing a substance to be measured (e.g., glucose) when bodily fluids containing saliva or blood are injected into the biosensor (112) through a collection device (111).

[0049] When the biosensor (112) is inserted into the measuring device (113), the measuring device (113) recognizes the insertion and switches to a standby state for recognizing bodily fluid injection by providing a specific signal to the biosensor (112). When bodily fluid is injected into the biosensor (112), the measuring device (113) recognizes that bodily fluid has been injected by the response signal (e.g., a change in resistance or voltage) generated by the biosensor (112). When it is determined that bodily fluid has been injected, the measuring device (113) applies a voltage for measuring the detection medium (e.g., glucose) to the working electrode and the reference electrode. Thus, based on the redox reaction between the detection medium and the reagent layer in the bodily fluid, a response signal (e.g., a response current) is generated from the biosensor (112), and the measuring device (113) receives this signal, thereby enabling the detection of the detection medium (e.g., glucose) or the measurement or prediction of the concentration of the detection medium.

[0050] Therefore, the measuring device (113) may include a memory, a processor, an input / output interface, etc.

[0051] Furthermore, the measuring device (113) may include a communication module to enable communication with the server (160) via a network (170). However, multiple electronic devices (110, 120, 130, 140, 150), each containing the measuring device (113), may not directly connect to the network (170) to communicate with the server (160), but may instead communicate via the network (170) with a terminal held by the subject (e.g., a smartphone, mobile phone, tablet computer, navigator, computer, laptop computer, digital broadcasting terminal, personal digital assistant (PDA), and portable multimedia player (PMP), or communicate with the server (160) via the terminal accessing the network (170).

[0052] The communication method is unrestricted and can include not only communication methods utilizing communication networks (e.g., mobile communication networks, wired Internet, wireless Internet, and broadcast networks) that can be included in the network (170), but also short-range wireless communication between devices. For example, the network (170) can include any one or more of the following networks: Personal Area Network (PAN), Local Area Network (LAN), Campus Area Network (CAN), Metropolitan Area Network (MAN), Wide Area Network (WAN), Broadband Network (BBN), and the Internet. In addition, the network (170) can include, but is not limited to, any one or more of the following network topologies: bus network, star network, ring network, mesh network, star-bus network, tree network, or hierarchical network.

[0053] Although the above describes the multiple electronic devices (110, 120, 130, 140, 150) as including the acquisition device (111), the biosensor (112), and the measuring device (113), it is not limited to this. As a measuring device (113), it may also include the terminal held by the person testing the object (e.g., smartphone, mobile phone, tablet computer, navigator, computer, laptop computer, digital broadcasting terminal, personal digital assistant (PDA), and portable multimedia player (PMP).

[0054] The server (160) may be implemented by a computer device or multiple computer devices that communicate with multiple electronic devices (110, 120, 130, 140, 150) via a network (170) to receive measured bodily fluid detection information from the respective measuring devices (113) of the multiple electronic devices (110, 120, 130, 140, 150) and provide a biometric measurement service for predicting the biometrics (e.g., blood glucose) of the subject based on the received bodily fluid detection information.

[0055] That is, multiple electronic devices (110, 120, 130, 140, 150) can each access the server (160) to receive biometric measurement services provided by 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 operating system).

[0056] Figure 2 This is a block diagram illustrating an example of a biosensor included in a biosensor system according to one embodiment. Figure 3 This is a diagram illustrating the structure of neutral red, which serves as an electron transport medium, in a reagent layer included in a biosensor according to one embodiment. Figures 4 to 5 This is a diagram illustrating the superior electron transport properties of neutral red, which serves as an electron transport mediator, in the reagent layer included in a biosensor according to one embodiment. Figure 6 This is a diagram illustrating the adjustment of the concentration of neutral red, which serves as an electron transport medium, in a reagent layer included in a biosensor according to one embodiment.

[0057] Reference Figure 2 The biosensor (200) may include a substrate (210), an electrode structure (220), and a reagent layer (230).

[0058] The substrate (210), which serves as the base for providing the electrode structure (220), can be formed of an insulating material to electrically separate the working electrode and the reference electrode contained in the electrode structure (220).

[0059] If the working electrode and the reference electrode are not formed on the same plane, but are formed independently on planes of different heights, the substrate (210) can be composed of multiple layers to accommodate the working electrode and the reference electrode.

[0060] The electrode structure (220) can be used to: promote the redox reaction between the detector in the body fluid and the reagent layer (230) through interaction with the detector and the reagent layer (230) in the body fluid, and send the current signal generated by the redox reaction to the measuring device.

[0061] That is, the electrode structure (220) is a component that includes a working electrode and a reference electrode and is provided to the substrate (210). Through this structure, the current generated by the reaction between the reagent layer (230) provided on the working electrode and the reference electrode and the detection body in the body fluid can be measured.

[0062] Therefore, the configuration and shape of the working electrode and the reference electrode can be implemented in a variety of ways, as long as they meet the conditions that allow the current generated by the reaction between the reagent layer (230) provided on the working electrode and the reference electrode and the detection body in the body fluid to be measured.

[0063] A reagent layer (230) is provided on the electrode structure (220) and may contain substances that undergo redox reactions with the body fluid injected onto the reagent layer (230). More specifically, the reagent layer (230) may include oxidoreductases and electron transport mediators.

[0064] Oxidoreductases can include glucose dehydrogenases (GDH). Specifically, oxidoreductases can 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, they are not limited to this, and oxidoreductases can include a variety of enzymes that undergo redox reactions with the body fluids.

[0065] Electron transport mediators may include ruthenium compounds. Specifically, ruthenium compounds may include at least one compound selected from the group consisting of hexaammineruthenium trichloride, ruthenium(III)hexaamine, ruthenium(II)tris(bipyridine) (2,2'-bipyridine), ruthenium(II)tris(4,4'-dicarboxy-2,2'-bipyridine) (4,4'-dicarboxy-2,2'-bipyridine) (4,4'-dicarboxy-2,2'-bipyridine) (ruthenium(III)ChloroDi(2,2'-bipyridine)). However, it is not limited to this, and the electron transfer medium may include various compounds used to mediate the rapid transfer of electrons generated by the reaction of the body fluid in the test body injected into the reagent layer (230) with the reagent layer (230) containing oxidoreductase to the electrode structure (220).

[0066] Electron transport mediators may further include neutral red. For example... Figure 3 As shown, neutral red can have 3-amino-7-dimethylamino-2-methylphenazine hydrochloride (C 15 H 17 The structure of ClN4).

[0067] Figures 4 to 6 The charts shown are based on the data described in Tables 1 through 3 below.

[0068] [Table 1]

[0069]

[0070] [Table 2]

[0071]

[0072] [Table 3]

[0073]

[0074] Thus, according to one embodiment, the biosensor (200) uses neutral red as an electron transport medium within the reagent layer (230), therefore... Figure 4As shown, unlike the case where neutral red is not used as an electron transport medium, the accuracy of the detector measurement can be improved by satisfying the condition that "the increase in current corresponding to the increase in detector concentration in the body fluid must be above the sensed rate of increase" based on the characteristic that the sensing current increases sharply according to the detector concentration.

[0075] Furthermore, according to one embodiment of the biosensor (200), instead of using electron transport mediators 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), neutral red is used as the electron transport mediator. Therefore, as Figure 5 As shown, compared to the use of electron transfer media such as DCIP, toluidine blue, NBT, and AQDS, this biosensor (200) exhibits the characteristic of a sharp increase in sensing current corresponding to the concentration of the detectable substance. Therefore, based on the characteristic of a sharp increase in sensing current corresponding to the concentration of the detectable substance, the biosensor (200) can improve the accuracy of detectable substance measurement by satisfying the condition that "the increase in current corresponding to an increase in the concentration of the detectable substance in the body fluid must be above a perceptible rate of increase".

[0076] Therefore, the concentration of neutral red used as an electron transport medium can be adjusted such that the current generated in response to the redox reaction between the detector and the oxidoreductase increases by a rate corresponding to the increase in the concentration of the detector in the body fluid, exceeding a predetermined increase rate.

[0077] More specifically, the current generated in response to the redox reaction between the detector and the oxidoreductase increases proportionally to the increase in the concentration of the detector in the body fluid, and also has the characteristic of increasing proportionally to the increase in the concentration of neutral red. For example, the current generated in response to the redox reaction between the detector and the oxidoreductase, such as... Figure 6As shown, the current increases proportionally with the increase in the concentration of the detector in the body fluid, and also proportionally with the increase in the concentration of neutral red. Specifically, the current generated in response to the redox reaction between the detector and the oxidoreductase is larger at a neutral red concentration of 1 mM than at a concentration of 0.1 mM.

[0078] Therefore, the current generated in response to the redox reaction between the detector and the oxidoreductase increases proportionally to the increase in detector concentration in the body fluid. That is, the increase in current is greater at a concentration of 1 mM than at a neutral red concentration of 0.1 mM.

[0079] A greater rate of increase in current implies greater ease of current sensing. Therefore, the concentration of neutral red can be adjusted such that the current generated in response to the redox reaction between the detector and the oxidoreductase increases at a rate corresponding to an increase in the concentration of the detector in the body fluid that reaches or exceeds a predetermined rate of increase (a critical rate of increase for accurate current sensing). For example, the concentration of neutral red can be adjusted to 0.1 mM such that the rate of increase in current reaches or exceeds a predetermined rate of increase (a critical rate of increase for accurate current sensing). More preferably, the concentration of neutral red can be adjusted to 1 mM such that the rate of increase in current reaches or exceeds a predetermined rate of increase (a critical rate of increase for accurate current sensing).

[0080] However, the concentration of neutral red is not limited to this and can also be adjusted such that the current generated in response to the redox reaction between the detector and the oxidoreductase reaches or exceeds a pre-set threshold (the minimum current value required for current sensing) relative to the reference concentration of the detector in the body fluid (the average concentration of the detector within the body fluid). This corresponds to the minimum conditions for the biosensor (200) to sense the target substance within the detector. For example, the concentration of neutral red can be adjusted to be above 0.1 mM and below 1 mM.

[0081] like Figure 6 As shown in the diagram and described in [Table 3], when the reagent layer contains 0.1 mM to 1 mM of neutral red, the current signal generated in response to the redox reaction between the detector and the oxidoreductase varies proportionally to the glucose concentration within a range of 357 nA to 3601 nA when the glucose concentration varies within a range of 40 mg / dL to 600 mg / dL. In this case, the rate of increase of the current signal based on the glucose concentration can be 3 nA / (mg / dL) to 6 nA / (mg / dL).

[0082] Figure 7This is a flowchart illustrating a method for manufacturing a biosensor according to one embodiment.

[0083] The manufacturing method described below refers to the manufacturing reference. Figures 1 to 6 The described biosensor process is based on the premise that it is executed by an automated and mechanized manufacturing system.

[0084] Reference Figure 7 In step 1 (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 that the electrode structure including the working electrode and the reference electrode is disposed and formed on the substrate.

[0085] In step 2 (S720), the manufacturing system may provide a reagent layer to the electrode structure. Providing a reagent layer to the electrode structure means configuring and forming a reagent layer on the electrode structure.

[0086] More specifically, in step 2 (S720), the manufacturing system can provide a reagent layer consisting of an oxidoreductase and a ruthenium compound as an electron transport medium, and neutral red.

[0087] Oxidoreductases can include glucose dehydrogenases (GDH). Specifically, oxidoreductases can 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, they are not limited to this, and oxidoreductases can include a variety of enzymes that undergo redox reactions with the body fluids.

[0088] Electron transport mediators may include ruthenium compounds. Specifically, a ruthenium compound may be at least one compound selected from the group consisting of hexaammineruthenium trichloride, ruthenium(III)hexaamine, ruthenium(II)tris(bipyridine) (2,2'-bipyridine), ruthenium(II)tris(4,4'-dicarboxy-2,2'-bipyridine) (4,4'-dicarboxy-2,2'-bipyridine) (4,4'-dicarboxy-2,2'-bipyridine) (ruthenium(III)ChloroDi(2,2'-bipyridine)). However, this is not the only possibility; the electron transport medium may include various compounds that mediate the rapid transfer of electrons generated by the reaction of a body fluid-containing detector injected onto the reagent layer with the reagent layer containing an oxidoreductase to the electrode structure.

[0089] Electron transport mediators may further include neutral red. For example... Figure 3 As shown, neutral red can have the structure of 3-amino-7-dimethylamino-2-methylphenazine hydrochloride (C15H17ClN4).

[0090] Specifically, in step 2 (S720), the manufacturing system adjusts the concentration of neutral red so that the current generated in response to the redox reaction between the detector and the oxidoreductase increases at a rate greater than a preset rate corresponding to the increase in the concentration of the detector in the body fluid, thereby enabling precise current sensing.

[0091] Furthermore, not limited to this, in step 2 (S720), the manufacturing system adjusts the concentration of neutral red to such that the current generated in response to the redox reaction between the detector and the oxidoreductase reaches a preset threshold above the reference concentration of the detector in the body fluid, thereby enabling current sensing of the detector at the reference concentration.

[0092] The apparatus including the measuring device (113) described above can be implemented by hardware components, software components, and / or combinations of hardware and software components. For example, the apparatus and components described in the embodiments can be implemented using one or more general-purpose or special-purpose computers, such as processors, controllers, arithmetic logic units (ALUs), digital signal processors (DSPs), microcomputers, field-programmable arrays (FPAs), programmable logic units (PLUs), microprocessors, or any other device capable of executing and responding to instructions. The processing device can execute an operating system and one or more software applications running on said operating system. Furthermore, the processing device can also access, store, manipulate, process, and generate data in response to the execution of software. For ease of understanding, although sometimes described as using a single processing device, those skilled in the art will understand that a processing device can include multiple processing elements and / or various types of processing elements. For example, a processing device can include multiple processors or a processor and a controller. In addition, other processing configurations, such as parallel processors, may also be included.

[0093] Software may include computer programs, code, instructions, or a combination thereof, and may configure a processing device to operate in a desired manner, or to issue instructions to the processing device independently or collectively. Software and / or data may be embodied in any type of machine, component, physical device, virtual equipment, computer storage medium, or device for interpretation by the processing device or for providing instructions or data to the processing device. Software may also be distributed across computer systems connected via a network, thereby being stored or executed in a distributed manner. Software and data may be stored on more than one computer-readable recording medium.

[0094] The method according to the embodiments can be executed by various computer means and implemented in the form of program instructions that can be recorded in a computer-readable medium. The computer-readable medium may contain program instructions, data files, and data structures, either individually or in combination. The program instructions recorded on the medium may be specifically designed and constructed for the embodiments, or may be those known and usable by those skilled in the art of computer software. Examples of computer-readable recording media 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 floppy disks; and hardware devices specifically constructed for storing and executing program instructions, such as ROMs (read-only memory), RAMs (random access memory), and flash memory. Examples of program instructions include not only machine language code generated by a compiler but also high-level language code that can be executed by a computer using an interpreter or the like.

[0095] While the embodiments have been described above with reference to limited examples and accompanying drawings, those skilled in the art can make various modifications and variations based on the above description. For example, even if the described techniques are performed in a different order than the described methods, or if the constituent elements of the described systems, structures, devices, and circuits are combined or integrated in a different manner than the described methods, or are replaced or substituted by other constituent elements or equivalents, appropriate results can still be achieved.

[0096] Therefore, other implementations, other embodiments, and contents equivalent to the claims are also within the scope of the following claims.

Claims

1. A biosensor for detecting glucose in body fluids using a measuring device, characterized in that, include: substrate; Multiple electrodes located on the substrate; as well as Reagent layer; and, The reagent layer includes: Oxidoreductases; and Electron transport mediators containing ruthenium compounds and neutral red; The biosensor is a biosensor that uses a measuring device to detect glucose in body fluids.

2. The biosensor for detecting glucose in body fluids using a measuring device according to claim 1, characterized in that, The concentration of the neutral red is above 0.1 mM and below 1 mM.

3. The biosensor for detecting glucose in body fluids using a measuring device according to claim 1, characterized in that, The electrode is used to transmit the current signal generated by the redox reaction between the glucose and the reagent layer to the measuring device. The concentration of glucose is determined based on the value of the current signal sent to the measuring device.

4. The biosensor for detecting glucose in body fluids using a measuring device according to claim 3, characterized in that, When the concentration of glucose varies within a range of 40 mg / dL to 600 mg / dL, the value of the current signal varies proportionally to the concentration of glucose within a range of 357 nA to 3601 nA.

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

6. The biosensor for detecting glucose in body fluids using a measuring device according to claim 1, characterized in that, The ruthenium compound is selected from at least one compound in the group consisting of hexaammineruthenium trichloride, ruthenium(III) hexaamine, ruthenium(II)tris(bipyridine) (2,2'-bipyridine), ruthenium(II)tris(4,4'-dicarboxy-2,2'-bipyridine) (4,4'-dicarboxy-2,2'-bipyridine) (4,4'-dicarboxy-2,2'-bipyridine) (ruthenium(III)ChloroDi(2,2'-bipyridine)).

7. The biosensor for detecting glucose in body fluids using a measuring device according to claim 1, characterized in that, The oxidoreductase is glucose dehydrogenase (GDH).

8. The biosensor for detecting glucose in body fluids using a measuring device according to claim 7, characterized in that, The oxidoreductase is selected from at least one compound 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).

9. A system for measuring glucose concentration in body fluids, characterized in that, include: Body fluid collection device; The biosensor according to any one of claims 1 to 8; and Measuring device; The system is used to measure the concentration of glucose in body fluids.

10. A method for manufacturing a biosensor for detecting glucose in body fluids, characterized in that, include: Steps for preparing the substrate; The step of forming a plurality of electrodes on the substrate; as well as The step of depositing a reagent layer on the electrode; and, The reagent layer includes: Oxidoreductases; and Electron transport mediators containing ruthenium compounds and neutral red; The manufacturing method is a method for manufacturing a biosensor for detecting glucose in body fluids.

11. The method for manufacturing a biosensor for detecting glucose in body fluids according to claim 10, characterized in that, The concentration of the neutral red is above 0.1 mM and below 1 mM.

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