Methods for manufacturing each of DNA / swcnt nanosensor, hydrogel composite sensor material comprising same, and three-dimensional nanosensor adhesive patch comprising hydrogel composite sensor material

Abstract

An embodiment of the present invention provides a method for manufacturing a DNA / SWCNT nanosensor, the method comprising the steps of: forming a mixed solution in which a single-walled carbon nanotube (SWCNT) and a single-stranded DNA (ssDNA) sequence are mixed; sonicating the formed mixed solution to adsorb the single-stranded DNA (ssDNA) sequence onto the surface of the single-walled carbon nanotube (SWCNT); and removing impurities and aggregates through centrifugation.

Description

A method for manufacturing each of a DNA / SWCNT nanosensor, a hydrogel composite sensor material including the same, and a three-dimensional nanosensor adhesive patch including the said hydrogel composite sensor material.

[0001] The present invention relates to a DNA / SWCNT nanosensor, a hydrogel composite sensor material containing the same, and a method for manufacturing each of a three-dimensional nanosensor adhesive patch containing said hydrogel composite sensor material. More specifically, the invention relates to a DNA / SWCNT nanosensor containing the same, a hydrogel composite sensor material containing the same, and a method for manufacturing each of a three-dimensional nanosensor adhesive patch containing said hydrogel composite sensor material, which includes a novel design strategy based on a biomimetic interface to enhance biocompatibility and stability beyond simple detection.

[0002] With the current advancement of nanosensor technology, the tracking of ions, small molecules, nucleic acids, lipids, and proteins has become essential for the non-invasive assessment and real-time monitoring of health status; effectively tracking these elements requires the management of microfluids containing the subjects of analysis and sensitive sensors to detect them.

[0003] Conventional mass spectrometry and chromatography are accurate, but they have limitations in the real-time analysis of small sample volumes.

[0004] To address these issues, nanosensors equipped with molecular recognition capabilities are emerging as a promising alternative, capable of providing real-time signals by utilizing optical, electrical, and mechanical properties.

[0005] However, when these are integrated into bulky devices, there is a problem in that analytical capabilities are limited across various sample volumes and fluid velocities.

[0006] To solve this, it is necessary to integrate a customized nano-probe into a 3D interface to enable efficient sample collection. To achieve this, (1) a new component capable of effectively absorbing and analyzing small amounts of fluid, (2) a technology to maintain stable contact with biological surfaces, and (3) a system capable of collecting irregular small amounts of fluid are required, and at the same time, there is a need to improve the efficiency and portability of the nanosensor.

[0007]

[0008] <Prior Art 1> Republic of Korea Registered Patent No. 1026468 B1

[0009]

[0010] The technical problem that the present invention aims to solve is to provide a DNA / SWCNT nanosensor including a novel design strategy that enhances biocompatibility and stability beyond simple sensing based on a biomimetic interface, a hydrogel composite sensor material including the same, and a method for manufacturing each of a 3D nanosensor adhesive patch including the said hydrogel composite sensor material.

[0011]

[0012] The technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned technical problems will be clearly understood by those skilled in the art to which the present invention belongs from the description below.

[0013]

[0014] To achieve the above technical objectives, one embodiment of the present invention provides a method for manufacturing a DNA / SWCNT nanosensor.

[0015] The method for manufacturing the DNA / SWCNT nanosensor according to one embodiment of the present invention is,

[0016] A method for manufacturing a DNA / SWCNT nanosensor may be characterized by comprising: a step of forming a mixed solution in which a single-walled carbon nanotube (SWCNT) and a single-stranded DNA (ssDNA) sequence are mixed; a step of sonicating the formed mixed solution to adsorb the single-stranded DNA (ssDNA) sequence onto the surface of the single-walled carbon nanotube (SWCNT); and a step of removing impurities and aggregates through centrifugation.

[0017]

[0018] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a DNA / SWCNT nanosensor characterized by performing a step of synthesizing single-walled carbon nanotubes (SWCNT) through a HiPCO (High-Pressure Carbon Monoxide) process prior to the step of forming the mixed solution.

[0019]

[0020] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a DNA / SWCNT nanosensor characterized in that the single-walled carbon nanotube (SWCNT) is a near-infrared (nIR) optical single-walled carbon nanotube (SWCNT).

[0021] In addition, according to one embodiment of the present invention, the step of forming the mixed solution is,

[0022] There may be a method for manufacturing a DNA / SWCNT nanosensor characterized by including single-walled carbon nanotubes (SWCNT) and single-stranded DNA (ssDNA) sequences in a mass ratio of 1:2 to 2:1 within the above-mentioned mixed solution.

[0023] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a DNA / SWCNT nanosensor, wherein the step of adsorbing the single-stranded DNA (ssDNA) sequence onto the surface of the single-walled carbon nanotube (SWCNT) is characterized by forming a corona phase of various structures on the surface of the single-walled carbon nanotube (SWCNT) during the process in which the single-stranded DNA (ssDNA) sequence is adsorbed onto the surface of the single-walled carbon nanotube (SWCNT), and enabling the detection of a specific substance by selectively binding to the corona phase using the molecular recognition principle of the corona phase.

[0024] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a DNA / SWCNT nanosensor characterized in that the single-stranded DNA (ssDNA) sequence is selected from the group consisting of (AC)15, (ACA)10, (ACCA)7, (ACG)10, (AG)15, (AGCA)7, (AGGA)7, (CACG)7, (CAGC)7, (CCCA)7, (CGCA)7, (GAAC)7, (GACG)7, (GAGC)7, (GCGA)7, (GGGC)7, (GT)15, (GTTG)7, and (TTTG)7.

[0025] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a DNA / SWCNT nanosensor characterized in that the specific substance is selected from the group consisting of Ascorbic acid, Thiamine, Riboflavin, Nicotinic acid, Pantothenic acid, Pyridoxine, Folic acid, Cortisol, Glucose, Ammonia, Urea, β-HB, L-Arginine, Leucine, and Creatine.

[0026]

[0027] To achieve the above technical problem, another embodiment of the present invention provides a method for manufacturing a hydrogel composite sensor material.

[0028] The method for manufacturing the hydrogel composite sensor material according to one embodiment of the present invention is,

[0029] A method for manufacturing a hydrogel composite sensor material may be characterized by comprising the steps of: providing a DNA / SWCNT nanosensor solution through the above-described DNA / SWCNT nanosensor manufacturing method; providing a polyacrylamide (PAAm) hydrogel; and mixing the DNA / SWCNT nanosensor solution with the polyacrylamide (PAAm) hydrogel.

[0030]

[0031] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a hydrogel composite sensor material, wherein the step of mixing the DNA / SWCNT nanosensor solution with a polyacrylamide (PAAm) hydrogel is characterized by mixing the DNA / SWCNT nanosensor solution and the polyacrylamide (PAAm) hydrogel in a volume ratio of 1:2 to 2:1.

[0032] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a hydrogel composite sensor material characterized in that the mixing step is carried out in a temperature range of 50°C to 70°C.

[0033]

[0034] To achieve the above technical problem, another embodiment of the present invention provides a method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid.

[0035] A method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid according to one embodiment of the present invention is,

[0036] A method for manufacturing a 3D nanosensor adhesive patch capable of absorbing and detecting microfluid may be characterized by comprising: a step of providing a hydrogel composite sensor material through the above method for manufacturing a hydrogel composite sensor material; a step of providing a 3D patch patterned so that a frog-mimicking hexagonal microstructure protrudes; a step of plasma surface treating the 3D patch; a step of filling the pattern of the 3D patch with the hydrogel composite sensor material; and a step of curing the 3D patch filled with the hydrogel composite sensor material.

[0037]

[0038] In addition, according to one embodiment of the present invention, the step of providing the three-dimensional patch comprises: filling a PDMS solution into a mold with a hexagonal pattern; heat-treating the mold filled with the PDMS solution to cure the PDMS solution; removing the mold to obtain a patterned PDMS with a protruding hexagonal microstructure; and forming an s-PDMS layer composed of PDMS and PEIE (Polyethyleneimine) on the surface of the protruding hexagonal microstructure of the PDMS; thereby providing a method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluids.

[0039]

[0040] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid, wherein, in the step of filling a PDMS solution into a mold of the hexagonal pattern, the PDMS is characterized by being a mixture of PDMS A and PDMS B in a volume ratio of 20:1.

[0041] In addition, according to one embodiment of the present invention, there may be a method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid, wherein the step of curing the PDMS solution is performed by heat-treating a mold filled with the PDMS solution at 50°C to 70°C.

[0042]

[0043] In addition, according to one embodiment of the present invention, the step of forming an s-PDMS layer composed of PDMS and PEIE (Polyethyleneimine) on the surface of a protruding hexagonal microstructure of the PDMS comprises: a step of depositing the s-PDMS composed of PDMS and PEIE (Polyethyleneimine) on a substrate; and a step of adhering the surface of the protruding hexagonal microstructure of the PDMS to the deposited s-PDMS and transferring it. There may be a method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid.

[0044] To achieve the above technical problem, another embodiment of the present invention provides a 3D nanosensor adhesive patch manufactured through a method for manufacturing a 3D nanosensor adhesive patch capable of absorbing and detecting microfluid.

[0045] In addition, a 3D multi-sensor adhesive patch capable of absorbing and detecting microfluid can be provided, characterized by arranging multiple 3D nanosensor adhesive patches, wherein the 3D patches include DNA / SWCNT nanosensors with different single-stranded DNA (ssDNA) sequences adsorbed thereon.

[0046]

[0047] According to one embodiment of the present invention, a DNA / SWCNT nanosensor including a novel design strategy that enhances biocompatibility and stability beyond simple detection based on a biomimetic interface, a hydrogel composite sensor material including the same, and a method for manufacturing each of a three-dimensional nanosensor adhesive patch including the said hydrogel composite sensor material can be provided.

[0048] According to one embodiment of the present invention, a nanosensor interface technology capable of detecting and analyzing minute fluid flows occurring on various biological surfaces can be provided.

[0049] According to one embodiment of the present invention, spatiotemporal data of molecules in an irregular fluid with ultra-low volume and low speed can be tracked and effectively analyzed in real-time and remotely through a three-dimensional nanosensor adhesive patch.

[0050]

[0051] The effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description of the invention or the claims.

[0052]

[0053] FIG. 1 is a schematic diagram briefly illustrating the molecular sensing mechanism of a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid according to one embodiment of the present invention.

[0054] Figure 2 is a schematic diagram showing the manufacturing process of a three-dimensional patch having a frog-mimicking hexagonal microstructure.

[0055] FIG. 3 is a schematic diagram of a DNA / SWCNT nanosensor in which DNA is adsorbed on the surface of a SWCNT according to one embodiment of the present invention.

[0056] FIG. 4 is a schematic diagram of (a) an absorbent polymer hydrogel composite sensor material combined with SWCNT according to one embodiment of the present invention, and (b) a flowchart showing the manufacturing process of a three-dimensional microstructure patch combined with the composite sensor material.

[0057] FIG. 5 is a schematic diagram of a three-dimensional multi-sensor adhesive patch in which a three-dimensional nanosensor adhesive patch according to one embodiment of the present invention is arranged in four multi-arrays, and an image of detecting an optical signal emitted by the multi-sensor adhesive patch in real time by absorbing a small amount of sweat from the surface of human skin.

[0058] FIG. 6 is an SEM image of a 3D nanosensor adhesive patch comprising (a) a biomimetic 3D microstructure patch and (b) a hydrogel composite sensor material combined with fluorescent SWCNTs according to one embodiment of the present invention, and (c) an actual photograph.

[0059] Figure 7 shows the fluid movement pattern at a representative interface height according to drainage modeling and the confocal fluorescence microscope at the corresponding stage, (a) is a schematic diagram of Stage I, where the fluid at the interface moves parallel, and Stage II, where it subsequently moves into a hexagonal channel within the microstructure, and (b) is data showing the fluid movement process observed through the confocal fluorescence microscope.

[0060] FIG. 8 is a schematic diagram and actual experimental image showing the process of absorbing a small amount (1.0 μL) of fluid over time of a 3D nanosensor adhesive patch according to one embodiment of the present invention and a comparative example.

[0061] FIG. 9 is data showing experimental results confirming the reactivity of each analyte to the sensor to evaluate the performance of a DNA / SWCNT nanosensor library according to one embodiment of the present invention.

[0062] FIG. 10 is the result data of a comparative measurement experiment of adhesive strength conducted by fabricating adhesive patches coated with various materials as a control group to verify the adhesive strength of a 3D nanosensor adhesive patch according to one embodiment of the present invention.

[0063] FIG. 11 is an image of the results of an in vitro experiment using an artificial skin model to evaluate the performance of a three-dimensional nanosensor adhesive patch according to one embodiment of the present invention.

[0064] FIG. 12 is data showing the experimental process and results conducted to test the reaction of a 3D nanosensor adhesive patch according to one embodiment of the present invention by simulating the sweat secretion rate in daily life at very slow flow rates of 0.5, 0.25, and 0.1 μL / min·cm².

[0065] FIG. 13 is the result data obtained by attaching the 3D nanosensor adhesive patch according to one embodiment of the present invention to the foreheads of three subjects and analyzing the sweat after consuming Vitamin C to verify the performance of the 3D nanosensor adhesive patch in an actual human body.

[0066]

[0067] The present invention will be described below with reference to the attached drawings. However, the present invention can be implemented in various different forms and is not limited to the embodiments described herein, and should be understood to include all modifications, equivalents, and substitutions that fall within the spirit and scope of the invention.

[0068] In addition, to clearly explain the invention in the drawings, parts unrelated to the explanation have been omitted, and similar parts throughout the specification have been given similar reference numerals.

[0069] Throughout the specification, when it is stated that a part is "connected (connected, in contact, joined)" to another part, this includes not only cases where they are "directly connected," but also cases where they are "indirectly connected" with other members in between.

[0070] Furthermore, when a part such as a layer, film, region, or plate is described as being “on” another part, this includes not only cases where it is “immediately above” the other part, but also cases where there is another part in between. Additionally, in this specification, when a part such as a layer, film, region, or plate is described as being formed “on” another part, the direction in which it is formed is not limited to the upward direction only, but includes cases where it is formed in the lateral or downward direction. Conversely, when a part such as a layer, film, region, or plate is described as being “below” another part, this includes not only cases where it is “immediately below” the other part, but also cases where there is another part in between.

[0071] In this specification, "upper surface" and "lower surface" are used as relative concepts to facilitate understanding of the technical concept of the present invention. Accordingly, "upper surface" and "lower surface" do not refer to specific directions, locations, or components, but are interchangeable.

[0072] For example, 'upper surface' may be interpreted as 'lower surface,' and 'lower surface' may be interpreted as 'upper surface.' Therefore, 'upper surface' may be expressed as 'No. 1' and 'lower surface' as 'No. 2,' or 'lower surface' may be expressed as 'No. 1' and 'upper surface' as 'No. 2.' However, within a single embodiment, 'upper surface' and 'lower surface' are not used interchangeably.

[0073] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by those skilled in the art to which the present invention pertains. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and should not be interpreted in an ideal or overly formal sense unless explicitly defined in this application.

[0074] Furthermore, when it is stated that a part "includes" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but rather allows for the inclusion of additional components.

[0075] The terms used herein are merely for describing specific embodiments and are not intended to limit the invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this specification, terms such as “comprising” or “having” are intended to indicate the presence of the features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, and should be understood as not precluding the existence or addition of one or more other features, numbers, steps, actions, components, parts, or combinations thereof.

[0076]

[0077] Advancements in nanosensor technology enable sophisticated sensing and are driving innovative changes in the fields of bioanalysis and diagnostics.

[0078] As the importance of rapid and accurate diagnosis grows in the fields of medicine and biotechnology, the need for technologies to extract useful information from minute amounts of fluid is becoming increasingly prominent; however, existing analytical techniques often make real-time diagnosis or on-site testing difficult due to the requirement of sufficient sample volumes and complex preprocessing steps.

[0079] Accordingly, the present invention presents a nanosensor interface technology capable of detecting and analyzing minute fluid flows occurring on various biological surfaces.

[0080] The above technology is based on a biomimetic interface and includes a new design strategy that goes beyond simple sensing to enhance biocompatibility and stability.

[0081] Through this, we propose a nanosensor-based interface and a method for fabricating it, which enable real-time diagnosis without complex electronic components and can make a significant contribution to the advancement of personalized medicine and precision medicine.

[0082]

[0083] The first objective of the present invention is to provide an adhesive drainage interface structure that mimics the hexagonal structural pattern of a frog's foot surface, capable of strongly adhering to rough and wet skin in dry and underwater environments while discharging surface fluid through channels.

[0084] To this end, the goal is to introduce a soft PDMS (Polydimethylsiloxane)-based microcoating layer on a hexagonal structure to exhibit strong adhesion through close contact with rough surfaces while maintaining stability at the biointerface.

[0085] Based on this, the present invention can provide a creative design and manufacturing method for a three-dimensional adhesive patch capable of controlling surface microfluidics optimized for dry or wet biological environments.

[0086]

[0087] The second objective of the present invention is to develop a method for manufacturing a nanosensor interface capable of stably bonding an absorbent polymer PAAm hydrogel composite sensing material combined with near-infrared fluorescent single-walled carbon nanotubes to the PDMS channel structure.

[0088] It provides a new interface that enables the capture and sensing of low-volume fluids on rough, dynamic biological surfaces, which were limited in conventional nanosensor interfaces, through simple surface treatment.

[0089] Based on sophisticated sensing capabilities, excellent adhesion, and surface fluid drainage performance, this can provide efficient molecular recognition and real-time monitoring of nanosensors for low-volume fluids.

[0090]

[0091] FIG. 1 is a schematic diagram briefly illustrating the molecular sensing mechanism of a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid according to one embodiment of the present invention.

[0092] Figure 2 is a schematic diagram showing the manufacturing process of a three-dimensional patch having a frog-mimicking hexagonal microstructure.

[0093] FIG. 3 is a schematic diagram of a DNA / SWCNT nanosensor in which DNA is adsorbed on the surface of a SWCNT according to one embodiment of the present invention.

[0094] FIG. 4 is a schematic diagram of (a) an absorbent polymer hydrogel composite sensor material combined with SWCNT according to one embodiment of the present invention, and (b) a flowchart showing the manufacturing process of a three-dimensional microstructure patch combined with the composite sensor material.

[0095] FIG. 5 is a schematic diagram of a three-dimensional multi-sensor adhesive patch in which a three-dimensional nanosensor adhesive patch according to one embodiment of the present invention is arranged in four multi-arrays, and an image of detecting an optical signal emitted by the multi-sensor adhesive patch in real time by absorbing a small amount of sweat from the surface of human skin.

[0096]

[0097] Embodiments of the present invention will be described in detail below with reference to the attached drawings.

[0098]

[0099] A method for manufacturing a DNA / SWCNT nanosensor according to the first embodiment of the present invention is described.

[0100]

[0101] As an example of the above embodiment, there may be a method for manufacturing a DNA / SWCNT nanosensor characterized by comprising: a step of forming a mixed solution in which a single-walled carbon nanotube (SWCNT) and a single-stranded DNA (ssDNA) sequence are mixed; a step of sonicating the formed mixed solution to adsorb the single-stranded DNA (ssDNA) sequence onto the surface of the single-walled carbon nanotube (SWCNT); and a step of removing impurities and aggregates through centrifugation.

[0102]

[0103] At this time, prior to the step of forming the above-mentioned mixed solution, the step of synthesizing single-walled carbon nanotubes (SWCNT) through a HiPCO (High-Pressure Carbon Monoxide) process may be performed first.

[0104]

[0105] In addition, the above single-walled carbon nanotube (SWCNT) may be a near-infrared (nIR) optical single-walled carbon nanotube (SWCNT).

[0106] During the process in which the above nanosensor binds to a specific analyte, the near-infrared (nIR) fluorescence intensity of the single-walled carbon nanotube (SWCNT) changes, and the target substance can be detected by measuring this.

[0107] Furthermore, since the degree of change varies depending on the concentration of the target substance, the amount of the target substance can also be verified.

[0108]

[0109] Referring to FIG. 3, it can be seen that single-stranded DNA (ssDNA) is adsorbed on the surface of the single-walled carbon nanotube (SWCNT).

[0110] The above single-stranded DNA (ssDNA) sequence is adsorbed onto the surface of the single-walled carbon nanotube (SWCNT) through ultrasonic treatment, and in this process, corona phases of various structures are formed on the surface of the single-walled carbon nanotube (SWCNT).

[0111] It can be confirmed that the substance can be detected by selectively binding to a specific substance using the molecular recognition principle of the corona described above.

[0112]

[0113] As an example of the above embodiment, the step of forming the mixed solution is,

[0114] There may be a method for manufacturing a DNA / SWCNT nanosensor characterized by including single-walled carbon nanotubes (SWCNT) and single-stranded DNA (ssDNA) sequences in a mass ratio of 1:2 to 2:1 within the above-mentioned mixed solution.

[0115] The above mass ratio is a factor related to detection performance, and most preferably, the mixed solution contains single-walled carbon nanotubes (SWCNT) and single-stranded DNA (ssDNA) sequences in a mass ratio of 1:1.

[0116]

[0117] The above DNA / SWCNT nanosensor targets human sweat, which has a very slow secretion rate and more than 100 constituent components.

[0118] Among the components of human sweat mentioned above, 15 challenging substances such as vitamins B and C, amino acids, and metabolic products can be detected.

[0119]

[0120] In the above DNA / SWCNT nanosensor, the single-stranded DNA (ssDNA) sequence adsorbed on the surface of the single-walled carbon nanotube (SWCNT) may be characterized as being any one selected from the group consisting of (AC)15, (ACA)10, (ACCA)7, (ACG)10, (AG)15, (AGCA)7, (AGGA)7, (CACG)7, (CAGC)7, (CCCA)7, (CGCA)7, (GAAC)7, (GACG)7, (GAGC)7, (GCGA)7, (GGGC)7, (GT)15, (GTTG)7, (TTTG)7.

[0121]

[0122] The above nucleotide sequence is written with the letters A, G, C, and T and numbers,

[0123] The above number indicates the number of times the alphabet inside the parentheses is repeated.

[0124] For example, (AC)15 means that AC is repeated 15 times, which is (ACACACACACACACACACACACACACACACAC).

[0125]

[0126] When the above-described single-stranded DNA (ssDNA) sequence is adsorbed onto the surface of the single-walled carbon nanotube (SWCNT), various corona phases of different structures are formed on the surface of the single-walled carbon nanotube (SWCNT) during the process, and

[0127] By utilizing the molecular recognition principle of the corona above, it selectively binds to any one selected from the group consisting of Ascorbic acid, Thiamine, Riboflavin, Nicotinic acid, Pantothenic acid, Pyridoxine, Folic acid, Cortisol, Glucose, Ammonia, Urea, β-HB, L-Arginine, Leucine, and Creatine, thereby finally enabling the detection of the corresponding substance.

[0128]

[0129] More preferably, in the DNA / SWCNT nanosensor, the single-stranded DNA (ssDNA) sequence adsorbed on the surface of the single-walled carbon nanotube (SWCNT) is preferably selected from the group consisting of (AC)15, (AGCA)7, (ACCA)7, and (ACA)10.

[0130] In this case, by utilizing the molecular recognition principle of the corona as described above, it becomes possible to finally detect any one selected from the group consisting of vitamin B2 (Riboflavin), vitamin B6 (Pyridoxine), vitamin B9 (Folic acid), and cortisol.

[0131] More specifically, (AC)15 / SWCNT selectively detects vitamin B2, (AGCA)7 / SWCNT selectively detects vitamin B6, (ACCA)7 / SWCNT selectively detects vitamin B9, and (ACA)10 / SWCNT selectively detects cortisol.

[0132] Through the combination of these various nanosensors, multiple biomarkers can be tracked simultaneously, which forms the basis of the multivariate molecular tracking capability of the 3D nanosensor adhesive patch described later.

[0133]

[0134] A method for manufacturing a hydrogel composite sensor material according to a second embodiment of the present invention is described.

[0135]

[0136] As an example of the above embodiment, there may be a method for manufacturing a hydrogel composite sensor material characterized by comprising: a step of providing a DNA / SWCNT nanosensor solution through the DNA / SWCNT nanosensor manufacturing method described above; a step of providing a polyacrylamide (PAAm) hydrogel; and a step of mixing the DNA / SWCNT nanosensor solution with the polyacrylamide (PAAm) hydrogel.

[0137]

[0138] That is, the above method for manufacturing the hydrogel composite sensor material contains the same technical concept as the above method for manufacturing the DNA / SWCNT nanosensor, so the content described above can be applied as is while explaining the above method for manufacturing the DNA / SWCNT nanosensor.

[0139]

[0140] The above PAAm hydrogel is a polymer material that absorbs a large amount of water to form a soft and flexible three-dimensional hydrophilic matrix,

[0141] Referring to FIG. 4a, the step of providing the PAAm hydrogel is,

[0142] It can be characterized by polymerizing acrylamide monomers using potassium persulfate as an initiator.

[0143] In Fig. 4a, the above KPS corresponds to potassium persulfate.

[0144] The above potassium persulfate is an initiator used in radical polymerization reactions, which decomposes easily in an aqueous solution to form radicals and thereby initiates a polymerization reaction that forms polymer chains.

[0145] When using the above potassium persulfate, there is an advantage in that the reaction rate and the length of the polymer chain can be controlled depending on the reaction conditions.

[0146]

[0147] As an example of the above embodiment, there may be a method for manufacturing a hydrogel composite sensor material in which the step of mixing the DNA / SWCNT nanosensor solution with a polyacrylamide (PAAm) hydrogel is characterized by mixing the DNA / SWCNT nanosensor solution and the polyacrylamide (PAAm) hydrogel in a volume ratio of 1:2 to 2:1.

[0148] More preferably, as an example of the above embodiment, there may be a method for manufacturing a hydrogel composite sensor material in which the step of mixing the DNA / SWCNT nanosensor solution with a polyacrylamide (PAAm) hydrogel is characterized by mixing the DNA / SWCNT nanosensor solution and the polyacrylamide (PAAm) hydrogel in a volume ratio of 1:1.

[0149] At this time, the mixing step may be carried out in a temperature range of 50℃ to 70℃ and for 15 to 30 minutes.

[0150]

[0151] A method for manufacturing a three-dimensional nanosensor adhesive patch (3D MIN) capable of absorbing and detecting microfluid according to the third embodiment of the present invention is described.

[0152]

[0153] As an example of the above embodiment, there may be a method for manufacturing a 3D nanosensor adhesive patch capable of absorbing and detecting microfluid, characterized by comprising: a step of providing a hydrogel composite sensor material through the above-described method for manufacturing a hydrogel composite sensor material; a step of providing a 3D patch patterned so that a frog-mimicking hexagonal microstructure protrudes; a step of plasma surface treating the 3D patch; a step of filling the pattern of the 3D patch with the hydrogel composite sensor material; and a step of curing the 3D patch filled with the hydrogel composite sensor material.

[0154] Referring to Fig. 4b, the above process can be seen as a schematic diagram.

[0155]

[0156] At this time, the three-dimensional patch patterned to protrude the frog-mimicking hexagonal microstructure consists of a protruding hexagonal microstructure and a channel which is a non-protruding part.

[0157] This can be confirmed through Fig. 6,

[0158] The width (w) of the channel, which is the non-protruding part, may be 180 μm to 220 μm, the height (h) of the channel may be 280 μm to 320 μm, and the spacing (s) between each channel may be 550 μm to 650 μm.

[0159]

[0160] In addition, the step of filling the pattern of the three-dimensional patch with the hydrogel composite sensor material may be carried out through the steps of: pouring the hydrogel composite sensor material onto the three-dimensional patch; and removing the overflowing portion.

[0161]

[0162] Referring to FIG. 2, as an example of the above embodiment, the step of providing the three-dimensional patch comprises: filling a PDMS solution into a mold with a hexagonal pattern; heat-treating the mold filled with the PDMS solution to cure the PDMS solution; removing the mold to obtain a patterned PDMS with a protruding hexagonal microstructure; and forming an s-PDMS layer composed of PDMS and PEIE (Polyethyleneimine) on the surface of the protruding hexagonal microstructure of the PDMS; thereby providing a method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid.

[0163]

[0164] At this time, in the step of filling the PDMS solution into the mold of the hexagonal pattern, the PDMS may be characterized as being a mixture of PDMS A and PDMS B in a volume ratio of 20:1.

[0165] In addition, the step of curing the PDMS solution can be carried out by heat-treating the mold filled with the PDMS solution at 50°C to 70°C, and the heat treatment can be carried out for 1.5 to 3 hours.

[0166]

[0167] As an example of the above embodiment, the step of forming an s-PDMS layer composed of PDMS and PEIE (Polyethyleneimine) on the surface of the protruding hexagonal microstructure of the PDMS comprises:

[0168] There may be a method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid, characterized by comprising: a step of depositing s-PDMS composed of PDMS and PEIE (Polyethyleneimine) on a substrate; and a step of transferring the protruding hexagonal microstructure surface of the PDMS to the deposited s-PDMS.

[0169]

[0170] At this time, the step of depositing s-PDMS composed of the above PDMS and PEIE (Polyethyleneimine) on a substrate can be carried out by spin coating, and the spin coating can be carried out at 500 rpm to 1000 rpm for 30 seconds to 90 seconds.

[0171]

[0172] In addition, the step of transferring the protruding hexagonal microstructure surface of the PDMS to the deposited s-PDMS can be carried out as a process of transferring for 5 seconds without external pressure.

[0173]

[0174] A three-dimensional nanosensor adhesive patch can be manufactured through the manufacturing method described above, and

[0175] Arrange a plurality of the above-mentioned 3D nanosensor adhesive patches,

[0176] The above 3D patch can also be manufactured to include DNA / SWCNT nanosensors with different single-stranded DNA (ssDNA) sequences adsorbed thereon, thereby enabling the 3D multisensor adhesive patch capable of detecting multiple target substances simultaneously.

[0177] Referring to Fig. 5, the structure of the three-dimensional multi-sensor adhesive patch can be seen in a schematic diagram.

[0178] Referring to Fig. 5, a 3D multisensor adhesive patch can be seen in which a total of four 3D nanosensor adhesive patches composed of (AC)15 / SWCNT, (AGCA)7 / SWCNT, (ACCA)7 / SWCNT, and (ACA)10 / SWCNT are arranged.

[0179]

[0180] Through the above 3D multisensor adhesive patch, various types of analytes can be measured simultaneously.

[0181] Although four arrays were demonstrated in Fig. 5 above, more three-dimensional nanosensor adhesive patch (3D MIN) units can be configured as a multi-array by adjusting the size of the substrate and the shape of each array unit.

[0182] Each 3D nanosensor adhesive patch (3D MIN) effectively captures the fluid into the hydrogel / sensor portion upon contact with the fluid thanks to a specially designed interface pattern, and the SWCNT sensor reacting with the captured fluid emits an optical signal in the near-infrared wavelength range.

[0183] The optical signal emitted by the above 3D nanosensor adhesive patch (3D MIN) is detected in real time through a near-infrared camera, and the intensity and spatial distribution of the signal can be determined simultaneously.

[0184]

[0185] The above 3D nanosensor adhesive patch mimics the structure of a frog's foreleg to guide fluid at the interface into the patch's channels, and by inserting a hydrogel composite sensor material combined with fluorescent SWCNTs having selective molecular recognition capabilities into the channels, it can efficiently absorb and precisely detect trace amounts of fluid.

[0186] In addition, it mimics the adhesive structure of a frog's forelegs, allowing for stable attachment even in humid environments or on rough surfaces, and has excellent effects in that it is biocompatible.

[0187]

[0188] Through the above 3D nanosensor adhesive patch, spatiotemporal data of molecules in an ultra-low volume, low-speed, irregular fluid can be tracked and analyzed effectively in real-time and remotely.

[0189] In addition, while tracking small amounts of analytes from the nanoscale to the microliter scale is very useful for precision medicine, early diagnosis, and the production of trace chemicals, traditional methods such as mass spectrometry or chromatography have made it difficult to analyze small amounts of rapidly evaporating samples in real time, whereas the 3D nanosensor adhesive patch (3D MIN) enables such analysis.

[0190]

[0191] In addition, the nanosensor equipped with the molecular recognition capability of the above-mentioned 3D nanosensor adhesive patch (3D MIN) can obtain real-time signals using optical, electrical, or mechanical properties, and can be a promising alternative to existing sensors, and

[0192] Through experiments described below, it was proven that reversible adhesion is possible without irritating human skin, thereby confirming the potential for actual application.

[0193]

[0194] Experimental Example 1. Structural analysis of a 3D nanosensor adhesive patch.

[0195]

[0196] Experimental Example 1 above is explained with reference to FIGS. 6 to 8.

[0197]

[0198] FIG. 6 is an SEM image of a 3D nanosensor adhesive patch comprising (a) a biomimetic 3D microstructure patch and (b) a hydrogel composite sensor material combined with fluorescent SWCNTs according to one embodiment of the present invention, and (c) an actual photograph.

[0199] Referring to Figures 6a and 6b, it can be seen that the surface of the protruding hexagonal structure of Figure 6a is coated with smooth s-PDMS.

[0200] Through the above configuration, stable bonding with the surface and high biocompatibility can be achieved.

[0201] The non-protruding channel portion drains microfluid present at the biological interface into the channel, and the hydrogel composite sensor material inserted into the channel absorbs the drained fluid and reacts with the captured fluid.

[0202]

[0203] Figure 7 shows the fluid movement pattern at a representative interface height according to drainage modeling and a confocal fluorescence microscope at the corresponding stage. (a) is a schematic diagram of Stage I, where the fluid at the interface moves parallel, and Stage II, where it subsequently moves into a hexagonal channel within the microstructure. (b) shows the fluid movement process observed through a confocal fluorescence microscope, allowing visual confirmation of the fluid height change and drainage through the microchannel at each stage.

[0204] At this time, the confocal fluorescence microscope was calibrated by setting the filter and laser to an absorption wavelength range of approximately 540-560 nm, and the lowest separation height in the fluid cross-section measured by the confocal microscope was confirmed using ImageJ software.

[0205] Referring to Fig. 7a, the fluid movement steps can be seen schematically.

[0206] Stage I is an initial stage in which the fluid is filled without hydrogel in a 3D MIN structure and moves parallel along the interface, and the separation height from the interface decreases from h0 to h1 and the fluid spreads flat.

[0207]

[0208] Stage II is a stage where the fluid is drained into the microstructure of the 3D MIN, and as the fluid is absorbed and drained into the hexagonal microchannel structure, the separation height is further reduced from h1 to h2. As a result, the fluid moves deeper through the channels and is absorbed.

[0209] Referring to Fig. 7b, one can see how the fluid separation height changes over time.

[0210] The curve (Modeling) is a theoretical modeling curve that predicts how the fluid separation height changes over time, and

[0211] The two points on the left (Radial squeeze) represent the translation phase in Stage I, which is the process in which the fluid reaches the separation height h1 through initial translation.

[0212] The two dots on the right (Drainage) represent the drainage phase in Stage II, showing the process of fluid draining along the hexagonal channel to reach the final separation height h2.

[0213]

[0214] FIG. 8 is a schematic diagram and actual experimental image showing the process of absorbing a small amount (1.0 μL) of fluid over time of a 3D nanosensor adhesive patch according to one embodiment of the present invention and a comparative example.

[0215] Referring to Fig. 8, the absorbent hydrogel composite sensor material inserted into the channel absorbs a small amount of fluid within about 3 seconds, whereas the 3D microstructure without hydrogel does not absorb a small amount of fluid even after 40 seconds.

[0216]

[0217] Experimental Example 2. Performance analysis of DNA / SWCNT nanosensor.

[0218]

[0219] Experimental Example 2 above is explained with reference to Fig. 9.

[0220]

[0221] FIG. 9 is data showing experimental results confirming the reactivity of each analyte to the sensor to evaluate the performance of a DNA / SWCNT nanosensor library according to one embodiment of the present invention.

[0222] Referring to Fig. 9a, the results of high-efficiency screening can be confirmed by fabricating SWCNT nanosensors functionalized with 20 different DNA sequences and reacting them with 15 target analytes.

[0223] The above experiment was performed using automated near-infrared spectroscopy, and a mixture of each nanosensor and analyte was reacted in a 96-well plate for 30 minutes and then excited with a 721 nm laser to collect fluorescence emission spectra in the 900-1400 nm range.

[0224] As a result of screening, each nanosensor exhibited a unique turn-on or turn-off response to a specific analyte, with response intensities ranging from -74% to 3,500%.

[0225]

[0226] In Fig. 9b, four representative nanosensors ((AC)15 / SWCNT, (AGCA)7 / SWCNT, (ACCA)7 / SWCNT, (ACA)10 / SWCNT) among the above nanosensors were selected and further characterization for vitamins B2, B6, B9, and cortisol was performed.

[0227] Referring to Fig. 9b, in order to evaluate the performance of the selected nanosensors in more detail, the response to analytes at various concentrations ranging from 1 nM to 1 μM was measured, and it can be confirmed that each nanosensor shows a dynamic increase or decrease in the near-infrared signal in proportion to the concentration of the target analyte.

[0228]

[0229] In Fig. 9c, a calibration curve was constructed and the limit of detection (LOD) was calculated based on the experimental results of Fig. 9b.

[0230] Referring to Fig. 9c, LOD values ​​of 1.74, 323, 991, and 2,270 nM were obtained for vitamins B2, B6, B9, and cortisol, respectively.

[0231]

[0232] The above experimental results demonstrate that the developed DNA / SWCNT nanosensor can detect various biomolecules with high selectivity and sensitivity.

[0233] In particular, it demonstrates that it operates effectively within the concentration range present in human sweat and possesses the ability to track multiple analytes simultaneously.

[0234] This is a core component of the 3D MIN system and serves as the foundation for enabling real-time multivariate molecular tracking.

[0235]

[0236] Experimental Example 3. Analysis of adhesion strength of a 3D nanosensor adhesive patch.

[0237]

[0238] Experimental Example 3 above is explained with reference to FIGS. 10 and FIGS. 11.

[0239]

[0240] FIG. 10 is the result data of a comparative measurement experiment of adhesive strength conducted by fabricating adhesive patches coated with various materials as a control group to verify the adhesive strength of a 3D nanosensor adhesive patch according to one embodiment of the present invention.

[0241]

[0242] In Fig. 10a, as comparison groups, a frog-mimicking adhesive patch (MIN-s) coated with s-PDMS on the surface of a hexagonal pad, a frog-mimicking adhesive patch (MIN-m) coated with a solution of PDMS A and B mixed in a 20:1 ratio on the surface of a hexagonal pad, a frog-mimicking adhesive patch (MIN-h) coated with a solution of PDMS A and B mixed in a 10:1 ratio on the surface of a hexagonal pad, and an adhesive patch having no microstructure were set.

[0243] In addition, as experimental conditions, three surfaces were prepared using flexible polyurethane acrylate (s-PUA) in a dry environment and a humid environment with sweat, with a flat surface and roughness (Ra) values ​​of 80 μm and 160 μm, and the adhesion strength was measured on these surfaces.

[0244] Referring to Figure 10a, the experimental results showed that the adhesive strength decreased as the surface roughness increased. MIN-s exhibited the highest adhesive strength and the lowest rate of decrease in adhesive strength on three surfaces in dry and wet environments. It was confirmed that the smooth coating layer applied to the surface increased the adhesive strength by closely contacting and filling the voids created on the rough surface.

[0245]

[0246] In Fig. 10b, the previous experimental group was attached to pig skin to measure the adhesion strength in different directions, and

[0247] Referring to Fig. 10b, experimental results show that MIN-s according to one embodiment of the present invention exhibits the highest shear stress (maximum ~13.3 kPa in dry environment, maximum ~11.3 kPa in wet environment) and high peel strength energy (maximum ~8.7 N / m in dry environment, maximum ~6.6 kPa in wet environment).

[0248]

[0249] In FIG. 10c, it can be seen that MIN-s according to one embodiment of the present invention is adhered to human skin and withstands a weight of 0.2 kg.

[0250]

[0251] FIG. 11 is an image of the results of an in vitro experiment using an artificial skin model to evaluate the performance of a three-dimensional nanosensor adhesive patch according to one embodiment of the present invention.

[0252] Referring to Fig. 11, it can be observed that skin irritation is minimized compared to other chemical adhesive-based adhesives when applied to the biological interface due to the biocompatible material properties of the 3D MIN's biomimetic 3D adhesive structure and the nanosensor / hydrogel.

[0253] Through this, it can be confirmed that the 3D nanosensor adhesive patch according to one embodiment of the present invention has a biomimetic structure designed to mimic the biological structure and adhesive function of nature, so it can be safely applied even for long-term use and has excellent biocompatibility that does not cause skin irritation.

[0254]

[0255] Experimental Example 4. Performance analysis of a 3D nanosensor adhesive patch.

[0256]

[0257] Experimental Example 4 above is explained with reference to FIGS. 12 and FIGS. 13.

[0258]

[0259] FIG. 12 is data showing the experimental process and results conducted to test the reaction of a 3D nanosensor adhesive patch according to one embodiment of the present invention by simulating the sweat secretion rate in daily life at very slow flow rates of 0.5, 0.25, and 0.1 μL / min·cm².

[0260] As can be seen in Fig. 12a, a test solution was supplied through a silicone tube with an inner diameter of 0.5 mm at a flow rate controlled by a syringe pump, and after attaching the 3D MIN to the artificial skin connected to the silicone tube, it was monitored in real time using a near-infrared microscope.

[0261]

[0262] In Fig. 12b, the response of 3D MIN according to the concentration of ascorbic acid (vitamin C) was measured.

[0263] Referring to Fig. 12b, when the ascorbic acid concentration was varied from 10 μM to 160 μM at a flow rate of 1.5 μL / min·cm², it can be seen that 3D MIN showed a continuous near-infrared signal increase response from 7% to 930%.

[0264] This means that 3D MIN operates stably over a wide concentration range.

[0265]

[0266] In Fig. 12c, the response of 3D MIN was tested at very slow flow rates of 0.5, 0.25, and 0.1 μL / min·cm² to simulate the sweat secretion rate in daily life.

[0267] Referring to Fig. 12c, it can be seen that 3D MIN can effectively capture fluid and detect a signal in 45.6 seconds even at an extremely slow flow rate of 0.1 μL / min·cm².

[0268] More specifically, while existing wearable sweat sensors require a sampling time of at least 100 seconds to 20 minutes and an average sampling volume of 1 μL, the 3D MIN according to one embodiment of the present invention analyzed a volume of only 75 nL in 45 seconds.

[0269]

[0270] FIG. 13 is the result data obtained by attaching the 3D multi-sensor adhesive patch according to FIG. 5 to the foreheads of three subjects and analyzing the sweat after consuming Vitamin C to verify the performance of the 3D nanosensor adhesive patch according to one embodiment of the present invention in an actual human body.

[0271] Figure 5 shows the responsiveness to various analogs by creating a multi-array of 3D MIN.

[0272] Referring to Fig. 5, it can be confirmed that the 3D MIN multi-array made of four arrays shows different unique signals for vitamins B2, B6, B9, and cortisol, thereby allowing the signal to be detected when reacting with a specific analyte.

[0273] In addition, in Fig. 13, to verify performance in an actual human body, a 3D multi-sensor adhesive patch according to Fig. 5 was attached to the foreheads of three subjects, and sweat analysis was performed after consuming Vitamin C.

[0274] At this time, the subjects consumed 3g of vitamin C and performed daily activities while taking measurements over 0, 50, 100, and 150 minutes.

[0275] Referring to FIG. 13b, 3D MIN according to one embodiment of the present invention successfully captured and analyzed minute amounts of sweat secreted from the forehead even when subjects were not exercising or using ion electrophoresis, and clearly showed changes in vitamin C concentration over time.

[0276] In particular, it demonstrates high spatial-temporal resolution by showing distinct near-infrared response pixels in different local regions even within a diameter of 1 cm.

[0277]

[0278] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single unit may be implemented in a distributed manner, and components described as distributed may likewise be implemented in a combined form.

[0279] The scope of the present invention is defined by the claims set forth below, and all modifications or variations derived from the meaning and scope of the claims and equivalent concepts thereof should be interpreted as being included within the scope of the present invention.

Claims

1. A step of forming a mixed solution in which single-walled carbon nanotube (SWCNT) and single-stranded DNA (ssDNA) sequences are mixed; A step of sonicating the above-formed mixed solution to adsorb the above-formed single-stranded DNA (ssDNA) sequence onto the surface of the above-formed single-walled carbon nanotube (SWCNT); and A method for manufacturing a DNA / SWCNT nanosensor characterized by including a step of removing impurities and aggregates through centrifugation.

2. In claim 1, prior to the step of forming the mixed solution, A method for manufacturing a DNA / SWCNT nanosensor characterized by performing a step of synthesizing single-walled carbon nanotubes (SWCNT) through a HiPCO (High-Pressure Carbon Monoxide) process prior to the process.

3. In claim 1, prior to the step of forming the mixed solution, A method for manufacturing a DNA / SWCNT nanosensor characterized by performing a step of synthesizing single-walled carbon nanotubes (SWCNT) through a HiPCO (High-Pressure Carbon Monoxide) process prior to the process.

4. In claim 1, the step of forming the mixed solution is, A method for manufacturing a DNA / SWCNT nanosensor characterized by including single-walled carbon nanotube (SWCNT) and single-stranded DNA (ssDNA) sequences in a mass ratio of 1:2 to 2:1 within the above-mentioned mixed solution.

5. In claim 1, the step of adsorbing the single-stranded DNA (ssDNA) sequence onto the surface of the single-walled carbon nanotube (SWCNT) comprises: During the process in which the above single-stranded DNA (ssDNA) sequence is adsorbed onto the surface of the above single-walled carbon nanotube (SWCNT), corona phases of various structures are formed on the surface of the above single-walled carbon nanotube (SWCNT), and A method for manufacturing a DNA / SWCNT nanosensor characterized by enabling the detection of a specific substance by selectively binding to it using the molecular recognition principle of the corona described above.

6. In paragraph 5, the single-stranded DNA (ssDNA) sequence is, A method for manufacturing a DNA / SWCNT nanosensor characterized by being selected from the group consisting of (AC)15, (ACA)10, (ACCA)7, (ACG)10, (AG)15, (AGCA)7, (AGGA)7, (CACG)7, (CAGC)7, (CCCA)7, (CGCA)7, (GAAC)7, (GACG)7, (GAGC)7, (GCGA)7, (GGGC)7, (GT)15, (GTTG)7, and (TTTG)7.

7. In paragraph 6, the specific substance mentioned above is, A method for manufacturing a DNA / SWCNT nanosensor characterized by using any one selected from the group consisting of ascorbic acid, thiamine, riboflavin, nicotinic acid, pantothenic acid, pyridoxine, folic acid, cortisol, glucose, ammonia, urea, β-HB, L-arginine, leucine, and creatine.

8. A step of preparing a DNA / SWCNT nanosensor solution through claim 1; A step of providing a polyacrylamide (PAAm) hydrogel; and A method for manufacturing a hydrogel composite sensor material characterized by including the step of mixing the above DNA / SWCNT nanosensor solution with the above Polyacrylamide (PAAm) hydrogel.

9. In claim 8, the step of mixing the DNA / SWCNT nanosensor solution with a polyacrylamide (PAAm) hydrogel is, A method for manufacturing a hydrogel composite sensor material characterized by mixing the above DNA / SWCNT nanosensor solution and the above Polyacrylamide (PAAm) hydrogel in a volume ratio of 1:2 to 2:

1.

10. A method for manufacturing a hydrogel composite sensor material according to claim 9, characterized in that the mixing step is carried out in a temperature range of 50℃ to 70℃.

11. A step of providing a hydrogel composite sensor material through claim 8; A step of providing a three-dimensional patch patterned so that a frog-mimicking hexagonal microstructure protrudes; A step of plasma surface treating the above three-dimensional patch; A step of filling the pattern of the above-described three-dimensional patch with the above-described hydrogel composite sensor material; and A method for manufacturing a 3D nanosensor adhesive patch capable of absorbing and detecting microfluid, characterized by including the step of curing a 3D patch filled with the above-mentioned hydrogel composite sensor material.

12. In claim 11, the step of providing the three-dimensional patch is, Step of filling a hexagonal pattern mold with PDMS solution; A step of curing the PDMS solution by heat-treating a mold filled with the above PDMS solution; A step of obtaining PDMS patterned such that a hexagonal microstructure protrudes by removing the above mold; and A method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid, characterized by including the step of forming an s-PDMS layer composed of PDMS and PEIE (Polyethyleneimine) on the surface of the protruding hexagonal microstructure of the PDMS.

13. A method for manufacturing a three-dimensional nanosensor adhesive patch capable of absorbing and detecting microfluid, wherein, in the step of filling a PDMS solution into a mold of the hexagonal pattern according to claim 12, the PDMS is characterized by being a mixture of PDMS A and PDMS B in a volume ratio of 20:

1.

14. In Clause 12, the step of curing the PDMS solution is, A method for manufacturing a 3D nanosensor adhesive patch capable of absorbing and detecting microfluid, characterized by heat-treating a mold filled with the above PDMS solution at 50°C to 70°C.

15. In claim 12, the step of forming an s-PDMS layer composed of PDMS and PEIE (Polyethyleneimine) on the surface of the protruding hexagonal microstructure of the PDMS comprises: A step of depositing s-PDMS composed of PDMS and PEIE (Polyethyleneimine) on a substrate; and A method for manufacturing a 3D nanosensor adhesive patch capable of absorbing and detecting microfluid, characterized by including the step of transferring the protruding hexagonal microstructure surface of the PDMS to the deposited s-PDMS.

16. A three-dimensional nanosensor adhesive patch manufactured according to Paragraph 11.

17. Arrange multiple 3D nanosensor adhesive patches according to Paragraph 16, wherein The above 3D patch is a 3D multisensor adhesive patch capable of absorbing and detecting microfluid, characterized by including DNA / SWCNT nanosensors with different single-stranded DNA (ssDNA) sequences adsorbed thereon.

Images