Structure for surface-enhanced raman scattering spectroscopy, manufacturing method therefor, and diagnosis method using same

Abstract

The present invention relates to a structure for surface-enhanced Raman scattering spectroscopy, a manufacturing method therefor, and a diagnosis method using same. With respect to the structure for surface-enhanced Raman scattering spectroscopy, the manufacturing method therefor, and the diagnosis method using same, according to the present invention, a structure for surface-enhanced Raman scattering spectroscopy and a manufacturing method therefor can be provided, the structure being capable of being used even if there is no biomarker having excellent diagnosis performance from among blood indexes, as with intractable cancers such as pancreatic cancers and intractable diseases. In addition, the diagnosis method and the diagnosis system, which use surface-enhanced Raman scattering spectroscopy as a structure for surface-enhanced Raman scattering spectroscopy, can be provided.

Description

Structure for surface-enhanced Raman scattering spectroscopy, method for manufacturing the same, and diagnostic method using the same

[0001] The present invention relates to a structure for surface-enhanced Raman scattering spectroscopy, a method for manufacturing the same, and a diagnostic method using the same.

[0002] Surface-enhanced Raman scattering (SERS) spectroscopy is a spectroscopic technique designed to complement Raman scattering spectroscopy, which has weak signals and low reproducibility. It is a spectroscopic technique in which the Raman scattering intensity of molecules adsorbed on the surface of metal nanostructures such as gold and silver increases rapidly by 10 6 ~ 10 8 It is a spectroscopy method that utilizes the phenomenon of increasing by more than a factor of two.

[0003] Surface-enhanced Raman scattering (SERS) spectroscopy is a technique that can obtain a large amount of information in a single measurement, is ultrasensitive enough to directly measure a single molecule, and can directly measure information about the vibrational state or molecular structure of a molecule, so it is recognized as a powerful analytical method for chemical / biological / biochemical analysis.

[0004] Recently, surface-enhanced Raman scattering spectroscopy has been attracting attention as a method for precisely obtaining comprehensive information on biomolecules from bioanalytical samples. However, the detection method for biomolecules using surface-enhanced Raman scattering spectroscopy proposed in the past requires specific binding, and there was a problem that screening and early diagnosis were difficult because there was no biomarker with excellent diagnostic performance among blood indicators for intractable cancers and intractable diseases such as pancreatic cancer.

[0005] Accordingly, there is a need for a diagnostic method and diagnostic system using surface-enhanced Raman scattering spectroscopy that can be used even when there is no biomarker with excellent diagnostic performance among biomarkers, such as intractable cancers and intractable diseases such as pancreatic cancer.

[0006] Meanwhile, the matters described as background technology are only intended to enhance understanding of the background of the present invention, and should not be taken as an acknowledgment that they correspond to prior art already known to those of ordinary skill in the art.

[0007] The technical problem to be achieved by the present invention is to provide a surface-enhanced Raman scattering spectroscopy structure and a method for manufacturing the same that can be used even when there is no biomarker with excellent diagnostic performance among blood indicators, such as intractable cancers and intractable diseases such as pancreatic cancer.

[0008] The technical problem to be achieved by the present invention is to provide a disease diagnosis method and diagnosis system using surface-enhanced Raman scattering spectroscopy that can be used even when a biomarker with excellent diagnostic performance among biomarkers does not exist.

[0009] In addition, the technical task to be achieved by the present invention is to provide a diagnostic method and diagnostic system using surface-enhanced Raman scattering spectroscopy having high repeatability and high convenience in distribution and storage.

[0010] The technical problems to be solved by the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0011] In order to achieve the above technical task, one embodiment of the present invention provides a structure for surface-enhanced Raman scattering spectroscopy, comprising: a silica shell layer having an accommodation space therein; and nanoparticles having a plasmonic metal, wherein the nanoparticles are arranged in one area of ​​the accommodation space within the silica shell layer.

[0012] In order to achieve the above technical task, another embodiment of the present invention provides a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, comprising the steps of: preparing a solution in which nanoparticles are dispersed; and forming a silica shell layer surrounding the nanoparticles dispersed in the solution.

[0013] In order to achieve the above technical task, one embodiment of the present invention provides a disease diagnosis method using surface-enhanced Raman scattering spectroscopy, including the steps of: obtaining a first surface-enhanced Raman scattering spectroscopy signal from a biological solution extracted from a patient with a disease; irradiating light on a diagnostic solution in which a metal nanostructure having a biomolecule material dispersed in a region therein is dispersed to obtain a second surface-enhanced Raman scattering spectroscopy signal in a region of the metal nanostructure; and comparing the first surface-enhanced Raman scattering spectroscopy signal and the second surface-enhanced Raman scattering spectroscopy signal to identify a disease of the patient with the disease.

[0014] In order to solve the above technical problem, another embodiment of the present invention provides a disease diagnosis system using surface-enhanced Raman scattering spectroscopy, including: a container in which a diagnostic solution is placed; an irradiation unit that irradiates light to the diagnostic solution placed in the container and generates a surface-enhanced Raman scattering spectroscopy signal in an area of ​​a metal nanostructure included in the diagnostic solution; and a diagnosis unit that analyzes the generated surface-enhanced Raman scattering spectroscopy signal.

[0015] According to one embodiment of the present invention, a surface-enhanced Raman scattering spectroscopy structure capable of non-specifically providing a surface-enhanced Raman scattering spectroscopy signal to a biomolecular material and a method for manufacturing the same can be provided.

[0016] In addition, according to one embodiment of the present invention, a surface-enhanced Raman scattering spectroscopy structure and a method for manufacturing the same can be provided that can be used even when there is no biomarker with excellent diagnostic performance among blood indicators, such as intractable cancers and intractable diseases such as pancreatic cancer.

[0017] According to one embodiment of the present invention, a diagnostic method and diagnostic system using surface-enhanced Raman scattering spectroscopy can be provided even if a biomarker with excellent diagnostic performance among biomarkers does not exist.

[0018] In addition, according to one embodiment of the present invention, a diagnostic method and diagnostic system using surface-enhanced Raman scattering spectroscopy having high repeatability and high convenience of distribution and storage can be provided.

[0019] 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 detailed description of the present invention or the composition of the invention described in the claims.

[0020] FIGS. 1A to 1E are conceptual diagrams showing a structure for surface-enhanced Raman scattering spectroscopy provided by one embodiment of the present invention.

[0021] FIG. 2a and FIG. 2b are conceptual diagrams showing a diagnostic solution for surface-enhanced Raman scattering spectroscopy provided by one embodiment.

[0022] FIG. 3a and FIG. 3b are flowcharts showing a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy provided by one embodiment.

[0023] FIG. 4a and FIG. 4b are flowcharts of a diagnostic method using surface-enhanced Raman scattering spectroscopy provided by one embodiment.

[0024] FIGS. 5A to 5C are block diagrams showing a diagnostic system provided by one embodiment.

[0025] FIGS. 6A and 6B are TEM images of a structure for surface-enhanced Raman scattering spectroscopy provided by one embodiment.

[0026] FIG. 7 is a diagram showing the results of a Raman scattering experiment when a biomaterial is present and when it is not present inside a structure according to one embodiment.

[0027] Figure 8 is a diagram showing the experimental results of differences in Raman spectra depending on whether ethanol is pretreated or not according to one embodiment.

[0028] Fig. 9 is a drawing confirming the surface-enhanced Raman scattering spectroscopy signal of a diagnostic solution provided by one embodiment.

[0029] <Explanation of symbols>

[0030] 1: Diagnostic solution

[0031] 10: Structure

[0032] 11: Nanoparticles

[0033] 12: Silica shell layer

[0034] 20: Dispersion

[0035] 100: Diagnostic System

[0036] 110: Courage Department

[0037] 111: Core

[0038] 112: Second plasmonic metal shell layer

[0039] 113: Polymer coating layer

[0040] 120: Investigation Department

[0041] 130: Diagnostic Department

[0042] 140: Data storage

[0043] One embodiment of the present invention provides a structure for surface-enhanced Raman scattering spectroscopy, comprising: a silica shell layer having an accommodation space therein; and nanoparticles having a plasmonic metal, wherein the nanoparticles are arranged in one area of ​​the accommodation space within the silica shell layer.

[0044] One embodiment of the present invention provides a surface-enhanced Raman scattering spectroscopy structure, wherein the nanoparticle includes a core including a first plasmonic metal and a second plasmonic metal shell layer disposed to surround the core and including a second plasmonic metal, and the second plasmonic metal shell layer has a symmetrical structure with respect to the center of the second plasmonic metal shell layer.

[0045] In addition, in one embodiment of the present invention, the structure may be a surface-enhanced Raman scattering spectroscopy structure including a biomolecular material disposed in the receiving space.

[0046] In addition, one embodiment of the present invention may be a surface-enhanced Raman scattering spectroscopy structure in which the first plasmonic metal and the second plasmonic metal each independently include at least one selected from the group consisting of Au, Ag, Cu, Al, W, Pt, Ni, and Pd.

[0047] In addition, one embodiment of the present invention may be a surface-enhanced Raman scattering spectroscopy structure in which the width of the plasmon resonance peak on the surface of the second plasmonic metal is narrower than the width of the plasmon resonance peak on the surface of the first plasmonic metal.

[0048] In addition, in one embodiment of the present invention, the second plasmonic metal shell layer may be a surface-enhanced Raman scattering spectroscopy structure having a regular polyhedral structure.

[0049] Another embodiment of the present invention can provide a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, comprising the steps of: preparing a solution in which nanoparticles are dispersed; and forming a silica shell layer surrounding the nanoparticles dispersed in the solution.

[0050] One embodiment of the present invention provides a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, wherein the step of preparing a solution in which the nanoparticles are dispersed includes a step of forming a mixed solution by mixing a first solution containing a first precursor compound containing a first plasmonic metal and a second solution containing a second precursor compound containing a second plasmonic metal, wherein the nanoparticles include a core containing the first plasmonic metal and a second plasmonic metal shell layer arranged to surround the core and having a second plasmonic metal, and wherein the second plasmonic metal shell layer has a symmetrical structure with respect to the center of the second plasmonic metal shell layer.

[0051] In addition, one embodiment of the present invention may be a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, further comprising a step of mixing a coating solution containing a water-soluble polymer compound into the mixed solution.

[0052] In addition, one embodiment of the present invention can provide a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, wherein the step of forming the silica shell layer includes the step of mixing the mixed solution, a third solution containing a biomolecular material, and a fourth solution containing a silica precursor compound, and the silica shell layer is formed so that at least a portion of the nanoparticles and the biomolecular material are arranged together inside.

[0053] Another embodiment of the present invention provides a disease diagnosis method using surface-enhanced Raman scattering spectroscopy, comprising: obtaining a first surface-enhanced Raman scattering spectroscopy signal from a biological solution extracted from a patient with a disease; irradiating light onto a diagnostic solution in which a surface-enhanced Raman scattering spectroscopy structure having a biomolecular material dispersed in a region therein is dispersed to obtain a second surface-enhanced Raman scattering spectroscopy signal in a region of the surface-enhanced Raman scattering spectroscopy structure; and comparing the first surface-enhanced Raman scattering spectroscopy signal and the second surface-enhanced Raman scattering spectroscopy signal to identify a disease of the patient with the disease.

[0054] In addition, one embodiment of the present invention may be a disease diagnosis method using surface-enhanced Raman scattering spectroscopy, wherein the surface-enhanced Raman scattering spectroscopy structure further includes a silica shell layer having an accommodation space inside; and nanoparticles having a plasmonic metal, wherein the nanoparticles and the biomolecule material are arranged in the accommodation space inside the silica shell layer.

[0055] In addition, one embodiment of the present invention may be a disease diagnosis method using surface-enhanced Raman scattering spectroscopy, wherein the biomolecular material includes two or more types of biomolecular materials.

[0056] In addition, one embodiment of the present invention may be a disease diagnosis method using surface-enhanced Raman scattering spectroscopy, wherein the diagnostic method further includes a step of creating a database of the first surface-enhanced Raman scattering spectroscopy signal obtained from a biological solution extracted from the patient with the disease.

[0057] In addition, one embodiment of the present invention may be a disease diagnosis method using surface-enhanced Raman scattering spectroscopy, wherein the diagnostic method further includes a step of obtaining a surface-enhanced Raman scattering spectroscopy signal for the biomolecular material and creating a database of information on the surface-enhanced Raman scattering spectroscopy signal for the biomolecular material.

[0058] Another embodiment of the present invention provides a disease diagnosis system using surface-enhanced Raman scattering spectroscopy, comprising: a container in which a diagnostic solution is placed; an irradiation unit that irradiates light to the diagnostic solution placed in the container and generates a surface-enhanced Raman scattering spectroscopy signal in an area of ​​a metal nanostructure included in the diagnostic solution; and a diagnosis unit that analyzes the generated surface-enhanced Raman scattering spectroscopy signal.

[0059] In one embodiment, the surface-enhanced Raman scattering spectroscopy structure may be a disease diagnosis system using surface-enhanced Raman scattering spectroscopy, wherein the structure further includes a silica shell layer having an accommodation space therein; and nanoparticles having a plasmonic metal, wherein the nanoparticles and the biomolecule material are arranged in the accommodation space within the silica shell layer.

[0060] Additionally, in one embodiment, the biomolecular material may be a disease diagnosis system using surface-enhanced Raman scattering spectroscopy, which includes two or more types of biomolecular materials.

[0061] In addition, in one embodiment, the diagnostic system may be a disease diagnostic system using surface-enhanced Raman scattering spectroscopy, further including a data storage unit that acquires the surface-enhanced Raman scattering spectroscopy signal and stores spectral information about the surface-enhanced Raman scattering spectroscopy signal.

[0062] Hereinafter, the present invention will be described with reference to the attached drawings. However, the present invention can be implemented in various different forms and is therefore not limited to the embodiments described herein. In the drawings, irrelevant parts have been omitted for clarity of description, and similar parts have been designated with similar reference numerals throughout the specification.

[0063] For convenience of explanation, the sizes of components in the drawings may be exaggerated or reduced. For example, the size and thickness of each component shown in the drawings may be arbitrarily indicated for convenience of explanation, and the following embodiments are not necessarily limited to those shown.

[0064] Terms such as first, second, A, and B may be used to describe various components, but these components should not be limited by these terms. These terms are used solely to distinguish one component from another. For example, without departing from the scope of the present invention, a first component may be referred to as a second component, and similarly, a second component may also be referred to as a first component. The term "and / or" includes a combination of multiple related items described herein or any of multiple related items described herein.

[0065] Throughout the specification, when a part is said to be "connected (connected, contacted, coupled)" to another part, this includes not only cases where it is "directly connected" but also cases where it is "indirectly connected" with another part in between.

[0066] Additionally, when a part is said to "include" a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise specifically stated.

[0067] The terminology used herein is for the purpose of describing specific embodiments only and is not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly dictates otherwise.

[0068] In this specification, terms such as “include” or “have” are intended to specify the presence of a feature, number, step, operation, component, part or combination thereof described in the specification, but should be understood not to exclude in advance the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts or combinations thereof.

[0069] When something is referred to as "on" or "on" another object, it includes not only the object directly above the other object, but also the case where there is an intervening object.

[0070]

[0071] Previously proposed methods for detecting biomolecules using surface-enhanced Raman scattering spectroscopy employed methods in which specific binding occurred between biomolecules and metal nanoparticles, resulting in the formation of surface-enhanced Raman scattering spectral signals at those locations. Consequently, even with surface-enhanced Raman scattering spectroscopy, it was difficult to use for diseases such as pancreatic cancer, where there were no biomarkers with high diagnostic performance.

[0072] FIGS. 1A to 1E are conceptual diagrams showing a structure for surface-enhanced Raman scattering spectroscopy provided by one embodiment of the present invention.

[0073] Referring to FIG. 1a, in order to solve the above-mentioned technical problem, one embodiment of the present invention provides a surface-enhanced Raman scattering spectroscopy structure (10) (hereinafter referred to as “structure”) including nanoparticles (11) and a silica shell layer (12), a surface-enhanced Raman scattering spectroscopy composition in which the structure (10) is dispersed, and a method for manufacturing the same.

[0074] In addition, as an optional embodiment, a biomolecular material (B) to be described later may be included inside the silica shell layer (12), and through the structure, a surface-enhanced Raman scattering spectroscopy signal can be obtained for a significant number of biomolecular materials (B) non-specifically arranged inside the structure (10) without excluding specific biomolecular materials (B).

[0075] Furthermore, by using a structure (10) like this and a composition for surface-enhanced Raman scattering spectroscopy in which these structures (10) are dispersed, by creating a database of surface-enhanced Raman scattering signals, it is possible to provide a diagnostic kit having excellent diagnostic performance even for diseases without a biomarker having excellent spectral performance.

[0076] Below, the structure (10) is described.

[0077] In one embodiment, the nanoparticle (11) may include a plasmonic metal. The surface-enhanced Raman scattering spectroscopy signal can be obtained by increasing the Raman scattering signal through a plasmon phenomenon at the local surface when molecules or particles exist near the surface of the plasmonic metal.

[0078] In this specification, “plasmonic metal” may mean a metal that is excited in a magnetic field, and examples thereof include Au, Ag, Cu, Al, W, Pt, Ni, and Pd.

[0079] In one embodiment, the nanoparticle (11) may include a core (111) including a first plasmonic metal, and a second plasmonic metal shell layer (112) arranged to surround the core (111) and including a second plasmonic metal. By making the nanoparticle (11) have a core-shell structure as described above, it is possible to provide a nanoparticle (11) having both properties of the first plasmonic metal and the second plasmonic metal.

[0080] In one embodiment, the second plasmonic metal shell layer (112) may have a symmetrical structure based on the center (M) of the second plasmonic metal shell layer (112). By having the second plasmonic metal shell layer (112) surround the core (111) and have a symmetrical structure as described above, the surface area between the nanoparticle (11) and the biomolecular material (B) can be increased, thereby maximizing the surface-enhanced Raman scattering spectral signal.

[0081] In one embodiment, the second plasmonic metal shell layer (112) may have a regular polyhedron structure, and the regular polyhedron may include, for example, at least one of a regular cube, a regular octahedron, a regular dodecahedron, and a regular icosahedron.

[0082] In one embodiment, the core (111) includes a first plasmonic metal, and the first plasmonic metal is any metal that can provide a surface-enhanced Raman scattering spectroscopy signal through a plasmon phenomenon on the surface, and should be interpreted as falling within the scope of the present invention, for example, Au, Ag, Cu, Al, W, Pt, Ni, and Pd, but is not limited to the above examples.

[0083] In one embodiment, the core (111) has an average particle diameter (D 50 ) can be made less than 50 nm. As another example, 0 to 50, 5 to 50, 10 to 50, 15 to 50, 20 to 50, 25 to 50, 30 to 50, 35 to 50, 40 to 50, 45 to 50, 0 to 45, 5 to 45, 10 to 45, 15 to 45, 20 to 45, 25 to 45, 30 to 45, 35 to 45, 40 to 45, 0 to 40, 5 to 40, 10 to 40, 15 to 40, 20 to 40, 25 to 40, 30 to 40, 35 to 40, 0 to 35, 5 to 35, 10 to 35, 15 to It can be 35, 20 to 35, 25 to 35, 30 to 35, 0 to 30, 5 to 30, 10 to 30, 15 to 30, 20 to 30, 25 to 30, 0 to 25, 5 to 25, 10 to 25, 15 to 25, or 20 to 25.

[0084] When the structures (10) are arranged in the dispersion (20) described later, having the above average particle size, an excellent Raman scattering effect can be provided.

[0085] In one embodiment, the core (111) may have a diameter of 10 nm to 50 nm in at least one direction. By having the above structure, when the structures (10) are arranged in the dispersion (20) described below, an excellent Raman scattering effect can be provided.

[0086] In one embodiment, the second plasmonic metal shell layer (112) includes a second plasmonic metal, and the width of the plasmon resonance peak on the surface of the second plasmonic metal can be narrower than the width of the plasmon resonance peak on the surface of the first plasmonic metal. As in the above structure, by making the width of the plasmon resonance peak on the surface of the second plasmonic metal narrower, when the structure (10) is dispersed in the dispersion (20) described below, better sensitivity can be provided when acquiring a Raman scattering signal.

[0087] At this time, as an optional example, the width of the plasmon resonance peak on the surface of the first plasmonic metal and the width of the plasmon resonance peak on the surface of the second plasmonic metal can be measured in the same environment.

[0088] In one embodiment, the second plasmonic metal shell layer (112) includes a second plasmonic metal, and the second plasmonic metal includes, but is not limited to, Au, Ag, Cu, Al, W, Pt, Ni, and Pd, for example.

[0089] In one embodiment, the core (111) may include gold (Au), and the second plasmonic metal shell layer (112) may include silver (Ag). Silver (Ag) has a light-gathering effect that is at least five times stronger than gold (Au), and a structure (10) including nanoparticles (11) having the above structure may provide an excellent surface-enhanced Raman scattering spectroscopy signal.

[0090] In one embodiment, the second plasmonic metal shell layer (112) may have a thickness of 10 nm to 70 nm in at least one direction. In another example, the thickness of the second plasmonic metal shell layer (112) may be 30 to 70 nm, 40 to 70 nm, 50 to 70 nm, 60 to 70 nm, 30 to 60 nm, 40 to 60 nm, or 50 to 60 nm. When the thickness of the second plasmonic metal shell layer (112) exceeds 70 nm, the size of the nanoparticles (11) arranged inside the structure (10) may become excessively large, and the reproducibility of surface-enhanced Raman scattering may deteriorate. When the thickness is less than 30 nm, the gap between the shell layer and the core (111) may become excessively narrow, and the reproducibility of surface-enhanced Raman scattering may deteriorate.

[0091] Referring to FIG. 1b, in one embodiment, the nanoparticles (11) may be formed on the outside of the second plasmonic metal shell layer (112) and may include a polymer coating layer (113) surrounding the second plasmonic metal shell layer (112). The polymer coating layer (113) imparts a negative charge to the surface of the nanoparticles (11) and may prevent the nanoparticles (11) from clumping together.

[0092] By having the above structure, in the manufacturing process of the structure (10), the nanoparticles (11) can be prevented from clumping together at a stage before forming the silica shell layer (12), and further, the silica shell layer (12) can be formed more easily.

[0093] In one embodiment, a type of surfactant, cetyltrimethylammonium chloride (CTAC), and a type of polymer, polyvinylpyrrolidone (PVP), may be applied to the surface of a metallic nanoparticle (11). CTAC can fix charged molecules to the surface of the nanoparticle (11) through electrostatic attraction, and PVP can fix various biomolecules to the surface through van der Waals attraction and chelation. This allows the CTAC and PVP applied to the surface of the metallic nanoparticle (11) to fix biomolecules in the blood to the surface of the nanoparticle during the process of forming the silica shell layer, thereby allowing the biomolecules in the blood to be easily located in the accommodation space within the silica shell layer.

[0094] In one embodiment, the polymer coating layer (113) is not particularly limited, and any polymer coating layer (113) that can be easily selected by a technician with ordinary knowledge in the art should be interpreted as falling within the scope of the present invention.

[0095] Meanwhile, the polymer coating layer (113) may include a water-soluble polymer compound. Examples of the water-soluble polymer compound include, but are not limited to, polyvinylpyrrolidone, polyvinyl alcohol, polyfluorosulfonate, hydroxyethyl cellulose, hydroxypropyl cellulose, cellulose acetate, and polyamide.

[0096] Referring to FIGS. 1c and 1d, in one embodiment, the structure (10) may further include a biomolecule material (B) disposed in the receiving space (H). By disposing the biomolecule material (B) together inside the silica shell layer (12) as in the above structure, the biomolecule material (B) disposed inside the receiving space (H) and the nanoparticles (11) can come into contact, and as described below, a surface-enhanced Raman scattering spectroscopy signal can be obtained from the surface of the nanoparticles (11).

[0097] In this specification, “biomolecular material” means a molecule and / or ion existing in a living organism, and may mean a molecule and / or ion synthesized and / or produced during a general biological process such as cell division, morphogenesis, or development.

[0098] Biomolecular substances (B) may include not only proteins, carbohydrates, lipids, nucleic acids, and minerals, but also small molecules such as primary metabolites, secondary metabolites, and natural products, and may include both endogenous and exogenous substances. Examples of biomolecular substances (B) include, but are not limited to, CA 19-9, amyloid beta40, and amyloid beta42 in plasma.

[0099] Meanwhile, as in the above structure, the structure (10) according to one embodiment may be provided with nanoparticles (11) that non-specifically cause a plasmon resonance phenomenon on the surface with biomolecular substances (B) to form a peak. In an optional embodiment, referring to FIG. 1e, different biomolecular substances (B) may be provided inside the structure (10). Through the above structure, multiple surface-enhanced Raman scattering spectroscopy signals can be obtained from even one structure (10).

[0100] Meanwhile, the acquired multiple surface-enhanced Raman scattering spectroscopy signals can be made into a database (DB) through multiple measurements, and based on the databased signals, specific peaks of diseases can be measured, and further, when used in diagnostic kits, etc., it can provide the effect of accurately diagnosing diseases in a short period of time.

[0101] In one embodiment, the structure (10) may include two or more types of biomolecular substances (B). By separating and extracting the upper or lower layer region of the biosolution treated with the pretreatment solution, the dimension of the total surface-enhanced Raman scattering spectroscopy signal obtained can be diversified, thereby enabling a more detailed database for a plurality of different biomolecular substances (B). Ultimately, a structure (10) capable of obtaining a surface-enhanced Raman scattering spectroscopy signal more accurately can be provided.

[0102] As described above, referring again to FIGS. 1A to 1E, in one embodiment, the structure (10) may include a silica shell layer (12). The silica shell layer (12) has an accommodation space (H) therein, and nanoparticles (11) and biomolecule materials (B) may be placed in an area within the accommodation space (H) inside the silica shell layer (12).

[0103] When nanoscale particles are dispersed in a liquid, etc., they may clump together due to interactions (e.g., attractive forces) between the particles within the dispersion (20). However, when a silica shell layer (12) surrounding individual nanoparticles (11) is provided as in the above structure, the silica shell layer (12) can prevent interactions between nanoparticles (11), thereby preventing agglomeration of the particles, and further, can provide an effect of stabilizing the measurement results by increasing the surface-enhanced Raman scattering spectroscopy signal or increasing the reproducibility of the surface-enhanced Raman scattering spectroscopy signal.

[0104] In addition, the structure (10) according to one embodiment does not necessarily require a substrate, location, or probe that specifically binds to biomolecular substances (B), and thus, when the biomolecular substances (B) are positioned on or come into contact with the surface of the nanoparticles (11), the surface-enhanced Raman scattering spectroscopy signals of a significant number of biomolecular substances (B) disposed inside the structure (10) can be measured.

[0105] As described above, the measured surface-enhanced Raman scattering spectroscopy signals can be databased, ultimately providing a diagnostic kit for diagnosing diseases even without a biomarker.

[0106] In one embodiment, the silica shell layer (12) may refer to a shell layer formed mainly with silicon (Si) and oxygen (O), and the material thereof is not particularly limited, and any silica shell layer (12) that can be easily selected by a technician with ordinary knowledge in the art should be interpreted as falling within the scope of the present invention. For example, there is a silica shell layer (12) formed by a polymerization reaction of TEOS, but the present invention is not limited to the above example.

[0107] In one embodiment, the silica shell layer (12) may have a thickness of 20 to 30 nm, and as another optional embodiment, 20 to 29, 20 to 28, 20 to 27, 20 to 26, 20 to 25, 20 to 24, 20.5 to 30, 20.5 to 29, 20.5 to 28, 20.5 to 27, 20.5 to 26, 20.5 to 25, 20.5 to 24, 21 to 30, 21 to 29, 21 to 28, 21 to 27, 21 to 26, 21 to 25, 21 to 24, 21.5 to 30, 21.5 to 29, 21.5 to 28, 21.5 to 27, 21.5 to 26, 21.5 to 25, 21.5 to 24, 22 to 30, 22 to 29, 22 to 28, 22 to 27, 22 to 26, 22 to 25, 22 to 24, 22.5 to 30, 22.5 to 29, 22.5 to 28, 22.5 to 27, 22.5 to 26, 22.5 to 25, 22.5 to 24, 23 to 30, 23 to 29, 23 to 28, 23 to 27, 23 to 26, 23 to 25, 23 to 24, 23.5 to 30, 23.5 to 29, It may have 23.5 to 28, 23.5 to 27, 23.5 to 26, 23.5 to 25, or 23.5 to 24.

[0108] Additionally, in an optional embodiment, the thickness of the silica shell layer (12) may be 23.28 nm or 21.85 nm, and this thickness may vary depending on whether or not ethanol pretreatment is performed.

[0109] Below, a composition for surface-enhanced Raman scattering spectroscopy is described.

[0110] FIG. 2a is a drawing showing a diagnostic solution used in a diagnostic method provided by one embodiment, and FIG. 2b is an enlarged drawing of area A of FIG. 2a.

[0111] Referring to FIG. 2a, another embodiment of the present invention provides a composition for surface-enhanced Raman scattering spectroscopy (hereinafter referred to as a “diagnostic solution”), which includes a dispersion (20) and a structure (10) for surface-enhanced Raman scattering spectroscopy (hereinafter referred to as a “structure”) dispersed within the dispersion (20). The composition can be used as a diagnostic solution (1) for surface-enhanced Raman scattering spectroscopy.

[0112] In one embodiment, a structure (10) may be dispersed in a dispersion (20) of a diagnostic solution (1), and a more specific and detailed description of the structure (10) is replaced with the description in the preceding embodiments.

[0113] In one embodiment, the dispersion (20) may refer to a liquid substance in which nanoscale particles can be dispersed. The dispersion (20) is not particularly limited, and any liquid substance capable of dispersing nanoparticles (11) that can be easily selected by a person skilled in the art is construed as falling within the scope of the present invention. Meanwhile, the dispersion (20) may include, for example, water (H2O), ethanol (EtOH), etc., but is not limited to the above examples.

[0114] In one embodiment, the diagnostic solution (1) may further include an additive. The additive may be, for example, a surfactant. The surfactant may enable the nanoscale particles and / or the aforementioned structures (10) to be more uniformly dispersed within the dispersion (20).

[0115] In one embodiment, the surfactant is not particularly limited, and cationic surfactants and / or anionic surfactants may be used. In addition, it should be understood that any surfactant that can be easily selected by a person skilled in the art is within the scope of the present invention. Meanwhile, surfactants include, for example, CTAB, CTAC, etc., and are not limited to the above examples.

[0116]

[0117] Below, a method for manufacturing a surface-enhanced Raman scattering spectroscopy structure (10) and a surface-enhanced Raman scattering spectroscopy diagnostic solution (1) is described.

[0118] FIG. 3a and FIG. 3b are flowcharts showing a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy provided by one embodiment.

[0119] Referring to FIG. 3a, in order to solve the above technical problem, one embodiment of the present invention provides a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy (hereinafter referred to as “structure manufacturing method”), which includes a step (S10) of forming a solution in which nanoparticles (11) are dispersed, and a step (S20) of forming a silica shell layer (12) surrounding the nanoparticles (11).

[0120] In addition, in an optional embodiment, the method for manufacturing a structure can perform the process of growing nanoparticles (11) in a solution and forming a silica shell layer (12) surrounding the nanoparticles (11), thereby providing a method for manufacturing a diagnostic solution (1) for surface-enhanced Raman scattering spectroscopy.

[0121] In one embodiment, the step (S10) of forming a solution in which nanoparticles (11) are dispersed may include a step of forming a mixed solution by mixing a first solution containing a first precursor compound including a first plasmonic metal and a second solution containing a second precursor compound including a second plasmonic metal.

[0122] By mixing the first precursor compound and the second precursor compound, the first plasmonic metal of the first precursor compound can grow into a core (111), and the second plasmonic metal of the second precursor compound can grow into a second plasmonic metal shell layer (112) along the outer surface of the core (111). Through this, core-shell nanoparticles (11) can be formed.

[0123] A more specific and detailed description of the first plasmonic metal and the second plasmonic metal is replaced by the description in the preceding examples.

[0124] Meanwhile, the step of forming core-shell nanoparticles (11) is not particularly limited, and any method that can be easily selected by a technician with ordinary knowledge in the art should be interpreted as falling within the scope of the present invention.

[0125] In one embodiment, the first precursor compound may refer to a compound that includes a first plasmonic metal and can form nanoscale nanoparticles including the first plasmonic metal. This is not particularly limited, and it should be interpreted that any precursor compound of nanoparticles that can be easily selected by a person skilled in the art is within the scope of the present invention. The first precursor compound may be, for example, HAuCl. 4· 3H2O exists, but is not limited to the above examples.

[0126] In one embodiment, the second precursor compound may refer to a compound that includes a second plasmonic metal and can form nanoscale nanoparticles or shells of nanoparticles including the second plasmonic metal. This is not particularly limited, and any precursor compound of a nanoparticle that can be easily selected by a person skilled in the art should be interpreted as falling within the scope of the present invention. Examples of the second precursor compound include AgNO3, but are not limited to the above examples.

[0127] In one embodiment, when mixing the first solution and the second solution, at least one of a reducing agent and a surfactant may be mixed together. When the mixed solution is formed, the first precursor compound forms a core (111) of a nanoparticle (11) through nanogrowth, and when at least one of a reducing agent and a surfactant is mixed together, the growth of the nanoparticle (11) can be further activated.

[0128] The reducing agent and surfactant are not particularly limited, and it should be interpreted that all reducing agents and surfactants that can be easily selected by a person skilled in the art are within the scope of the present invention, and examples thereof include NaBH4, CTAB, and CTAC, respectively.

[0129] In one embodiment, the method for manufacturing a structure may further include a step of mixing a mixed solution and a coating solution containing a water-soluble polymer compound. When the water-soluble polymer compound is mixed into the mixed solution containing nanoparticles (11), the water-soluble polymer compounds form a polymer coating layer (113) on at least one area on the surface of the nanoparticles (11), thereby preventing the nanoparticles (11) from agglomerating with each other.

[0130] In one embodiment, the water-soluble polymer compounds include, but are not limited to, polyvinylpyrrolidone, polyvinyl alcohol, polyfluorosulfonate, hydroxyethyl cellulose, hydroxypropyl cellulose, cellulose acetate, and polyamide.

[0131] The polymer coating layer (113) also enables biomolecules to be easily positioned on the metal surface, thereby providing superior sensitivity when acquiring Raman signals. For example, blood contains various biomolecules such as nucleic acids, proteins, organic substances, and inorganic substances, and the polymer coating layer (113) can be applied to the surface of the nanoparticles and act as an intermediate substance that can form molecular interactions between the nanoparticles and biomolecules so that these biomolecules can be positioned between the metallic nanoparticles and the silica shell layer.

[0132] Biomolecules in the blood can be attached to the intermediate substance applied to the surface of the metallic nanoparticles through intermolecular interactions, and the process of forming a silica shell layer on the surface where no biomolecules are attached begins, and in the process of wrapping the rapid nanoparticles, the biomolecules are fixed without being detached, and as a result, various biomolecules in the blood can exist in the accommodation space between the metallic nanoparticles and the silica shell layer.

[0133] Meanwhile, in order to form a polymer coating layer (113) on the surface of the nanoparticle (11), as an optional example, a mixed solution may be formed and the coating solution may be mixed after a predetermined period of time.

[0134] In one embodiment, the method for manufacturing a structure may further include a step of mixing a coating solution comprising a polymer compound comprising a unit represented by the following chemical formula 1 into the mixed solution, and as an optional embodiment, may further include a step of mixing a coating solution comprising a water-soluble polymer compound comprising a unit represented by the following chemical formula 1:

[0135] [Chemical Formula 1]

[0136]

[0137] In one embodiment, the method for manufacturing a structure may include a step of mixing a solution or mixed solution in which nanoparticles (11) are dispersed, a third solution containing a biomolecule material (B), and a fourth solution containing a silica precursor compound. By mixing and reacting the biomolecule material (B) and the silica precursor compound in the solution containing nanoparticles (11), the nanoparticles (11) and the biomolecule material (B) can be arranged together inside the silica shell layer (12).

[0138] In one embodiment, the biomolecule material (B) and the silica precursor compound can be mixed and reacted in a solution containing nanoparticles (11) using a stirrer, and as an optional example, the mixture can be mixed and reacted in an orbital shaker or a seesaw shaker.

[0139] Other more specific and detailed descriptions of the biomolecular material (B) are replaced with the descriptions in the preceding examples.

[0140] In one embodiment, the silica precursor compound may refer to compounds capable of forming a silica shell layer (12) after reaction, and this is not particularly limited, and any silica precursor compound that can be easily selected by a person skilled in the art should be interpreted as falling within the scope of the present invention. Examples of these include, but are not limited to, TEOS.

[0141] Referring to FIG. 3B, in one embodiment, the third solution containing the biomolecule material (B) may be a solution treated with a pretreatment solution (S15). The pretreatment solution may contain substances with low polarity, and as an optional example, may contain substances with a lower polarity than water.

[0142] The third solution can be prepared by treating and / or processing a solution directly extracted from a living organism (hereinafter referred to as a "biological solution"). Examples of biological solutions include plasma, tissue fluid, and urine.

[0143] These biosolutions can use water (H2O) as a solvent, and when a pretreatment solution containing a substance with low polarity is mixed with the biosolution, biomolecule substances (B) with different molecular weights in the biosolution can be separated according to molecular weight.

[0144] In one embodiment, the substance having a lower polarity than water includes, but is not limited to, aprotic substances, methanol, ethanol, acetone, DMSO, etc., but any substance having a lower polarity than water that can be easily selected by a person skilled in the art should be interpreted as falling within the scope of the present invention.

[0145] In one embodiment, the third solution may include two or more types of biomolecule substances (B).

[0146] By separating and extracting the upper or lower region of a biological solution treated with a pretreatment solution, the dimension of the total surface-enhanced Raman scattering spectroscopy signal obtained can be diversified, enabling a more detailed database of biomolecular substances (B). Ultimately, a structure (10) capable of diagnosing diseases more accurately can be provided.

[0147] It goes without saying that the embodiments of the present invention described above can be applied in combination with each other.

[0148]

[0149] In another aspect, the present invention relates to a diagnostic method and a diagnostic system using surface-enhanced Raman scattering spectroscopy.

[0150] FIG. 4a and FIG. 4b are flowcharts of a diagnostic method using surface-enhanced Raman scattering spectroscopy provided by one embodiment.

[0151] Referring to FIG. 4a, in order to solve the above technical problem, one embodiment of the present invention can provide a diagnostic method using surface-enhanced Raman scattering spectroscopy (hereinafter referred to as “the present diagnostic method”), which includes an irradiation step (S100) of irradiating light to a diagnostic solution (1) containing a biomolecule material (B) to obtain a surface-enhanced Raman scattering spectroscopy signal, and a step (S200) of analyzing the obtained surface-enhanced Raman scattering spectroscopy signal to diagnose a target disease.

[0152] Within a living organism with a specific disease, biomolecular substances that appear specifically for that specific disease may exist. A diagnostic method for diagnosing a target disease can be provided using specific surface-enhanced Raman scattering spectral signals (hereinafter referred to as "spectral signals") obtained from such biomolecular substances.

[0153] As described below, a diagnostic method according to one embodiment may include a step (not shown) of acquiring a first spectral signal from a biological solution extracted from a patient suffering from a disease or a solution containing the same. In addition, by acquiring first spectral signals from patients suffering from various diseases, spectral information on specific spectral signals according to the type of disease may be stored in a data storage unit (140) described below.

[0154] Target diseases include, but are not limited to, pancreatic cancer, Alzheimer's, rheumatism, corona, influenza, Alzheimer's dementia, etc.

[0155] In one embodiment, the biological solution refers to a liquid existing within a human body, such as blood, plasma, tissue fluid, lymph, urine, etc. In addition, the present invention is not limited to the above examples, and any liquid within the human body that can be easily extracted by a general public, a technician with ordinary knowledge in the art, a medical professional, or a medical assistant should be interpreted as falling within the scope of the present invention.

[0156] A diagnostic method according to one embodiment may include an irradiation step (S100) of irradiating a diagnostic solution (1) containing a biomolecular material (B) with light to obtain a spectral signal. As described above, such a diagnostic solution may have structures for surface-enhanced Raman scattering spectroscopy dispersed therein, and may use a composition for surface-enhanced Raman scattering spectroscopy in which the structure (10) for surface-enhanced Raman scattering spectroscopy (hereinafter referred to as “structure”) is dispersed. In addition, such diagnostic solutions may be transported in a produced state, or may be manufactured at a location where a disease is diagnosed. A more specific and detailed description of the diagnostic solution will be replaced with the description in the preceding embodiments in relation to the structure (10) and the composition in which the structure (10) is dispersed.

[0157] In one embodiment, a second spectral signal can be obtained by irradiating light on a diagnostic solution (1) containing a biomolecular substance (B). At this time, as will be described later, by comparing and analyzing the second spectral signal with spectral information stored in a data storage unit (140) or the like, it is possible to diagnose whether the diagnostic solution is for a patient with a certain disease. In addition, as an optional embodiment, during the comparative analysis process as described above, the biomolecular substances (B) dissolved or dispersed in the diagnostic solution can be identified.

[0158] As described above, the present diagnostic method provided by one embodiment of the invention obtains a spectral signal by directly irradiating light on a liquid diagnostic solution, and since a separate drying process is not required, the time required for diagnosis can be drastically reduced.

[0159] In addition, the diagnostic method provided by one embodiment diagnoses diseases, etc. by directly irradiating light (L) onto a liquid diagnostic solution (1), so that the problem of contamination of the experimental environment may not occur significantly compared to existing diagnostic methods. Furthermore, the structures (10) dispersed in the liquid, which will be described later, can maintain a similar environment even during repeated experiments, thereby increasing reproducibility.

[0160] In addition, since the diagnostic solution (1) used in the present diagnostic method provided by one embodiment can be stored and transported in a liquid state, convenience of storage and transportation of the diagnostic solution (1) can be secured. In addition, since it is manufactured in a liquid state, mass production of the diagnostic solution (1) is also possible.

[0161] In one embodiment, the light (L) irradiated to the diagnostic solution (1) may be light having a wavelength and intensity used for surface-enhanced Raman scattering or Raman scattering, and examples thereof include laser light. However, the present invention is not limited to the above examples.

[0162] Referring to FIG. 2a, in one embodiment, the diagnostic solution (1) may include a dispersion (20) and a structure (10) dispersed within the dispersion (20). As in the above structure, in the present invention, a spectral signal can be obtained by irradiating light (L) to the structure (10) dispersed in a liquid state without drying the diagnostic solution (1), thereby increasing the reproducibility and accuracy of the signal.

[0163] In addition, the diagnostic solution (1) can be manufactured by mixing with a biological solution extracted from a patient suffering from a disease, thereby allowing the biomolecular substances (B) contained in the structure (10) to be included in the diagnostic solution (1).

[0164] Referring to FIG. 2b, the structure (10) may include a biomolecule material (B) in one area inside, and when light is irradiated between the biomolecule material (B) arranged inside the structure (10) and one area of ​​the structure (10), a surface plasmon phenomenon occurs, and a specific second spectral signal (S) according to the type of biomolecule material (B) may be generated in one area of ​​the structure (10). These second spectral signals (S) can be compared and analyzed with the above-described spectral information to diagnose a target disease.

[0165] In addition, referring again to FIG. 2b, different biomolecular substances (B1 to B4) may be included in a region within the structure (10). In this way, different biomolecular substances (B1 to B4) generate different spectral signals (S1 to S4). In addition, the spectral signals (S1 to S4) generated at this time are spectral signals that are specifically generated from the biomolecular substances (B1 to B4).

[0166] By comparing and analyzing spectral signals with the aforementioned spectral information, target diseases can be diagnosed. Furthermore, as an optional example, biomolecular substances (B1 to B4) can be identified by analyzing these spectral signals (S1 to S4).

[0167] By providing a silica shell layer (12) as in the structure of the structure (10) of FIGS. 1a to 1e, the structures (10) can be well dispersed within the dispersion (20) without clumping together. In addition, by arranging nanoparticles (11) having a plasmonic metal and a biomolecule material (B) together within the silica shell layer (12), a surface-enhanced Raman scattering spectroscopy signal can be generated more efficiently between the nanoparticles (11) and the biomolecule material (B).

[0168] Referring again to FIGS. 1E and 2B, in one embodiment, the biomolecule material (B) may include two or more types of biomolecule materials (B1 to B4), and when different types of biomolecule materials (B) are included, specific surface-enhanced Raman scattering spectroscopy signals (S1 to S4) corresponding to the biomolecule materials (B) may be generated.

[0169] In one embodiment, the drawing illustrates the second plasmonic metal shell layer (112) as having a cubic shape, but the second plasmonic metal shell layer (112) may have a regular polyhedron structure in addition to a regular cubic shape, and the regular polyhedron may include, for example, at least one of a regular octahedron, a regular dodecahedron, and a regular icosahedron.

[0170] In one embodiment, the second plasmonic metal shell layer (112) includes a second plasmonic metal, and the width of the plasmon resonance peak on the surface of the second plasmonic metal can be narrower than the width of the plasmon resonance peak on the surface of the first plasmonic metal.

[0171] As in the above structure, by making the width of the peak of the plasmon resonance on the surface of the second plasmonic metal narrower, when the structure (10) is dispersed in the dispersion (20), it is possible to provide better sensitivity when obtaining a surface-enhanced Raman scattering spectroscopy signal.

[0172] At this time, as an optional example, the width of the plasmon resonance peak on the surface of the first plasmonic metal and the width of the plasmon resonance peak on the surface of the second plasmonic metal can be measured in the same environment.

[0173] Referring to FIGS. 1A to 1E, in one embodiment, the nanoparticle (11) may include a core (111), a second plasmonic metal shell layer (112), and a polymer coating layer (113) surrounding the second plasmonic metal shell layer (112). The polymer coating layer (113) imparts a negative charge to the surface of the nanoparticles (11) and may prevent the nanoparticles (11) from clumping together.

[0174] By having the above structure, in the manufacturing process of the structure (10), the nanoparticles (11) can be prevented from clumping together at a stage before forming the silica shell layer (12), and further, the silica shell layer (12) can be formed more easily. A more detailed description of the structure (10) is replaced with the description in the related preceding examples.

[0175] In one embodiment, the structure (10) can be formed by mixing and reacting a solution in which nanoparticles (11) are dispersed, a solution containing a biomolecule material (B), and a solution containing a silica precursor compound. A more detailed description of the method for manufacturing the structure (10) is replaced with the description in the related preceding embodiments.

[0176] In one embodiment, the solution containing the biomolecule substance (B) may be, for example, a biological solution extracted from a living organism, or may be a solution in which the biomolecule substance (B) is mixed with a solvent, but is not limited to these examples. Meanwhile, the solution containing the biomolecule substance (B) may include a pretreated solution, and the solution used in such a pretreatment process may be a solution having a lower polarity than water.

[0177] By performing a preprocessing process like this, the solution containing the biomolecular substance (B) can have various chemical or physical environments, and the dimension of the total surface-enhanced Raman scattering spectroscopy signal obtained can be diversified, thereby enabling a more detailed database of the biomolecular substance (B). Ultimately, a structure (10) capable of obtaining a surface-enhanced Raman scattering spectroscopy signal more accurately can be provided.

[0178] In one embodiment, the substance having a lower polarity than water includes, but is not limited to, aprotic substances, methanol, ethanol, acetone, DMSO, etc., but any substance having a lower polarity than water that can be easily selected by a person skilled in the art should be interpreted as falling within the scope of the present invention.

[0179] In one embodiment, the method for preparing the structure (10) is not particularly limited, and it should be interpreted that all methods for preparing the structure (10) that can be easily selected by a technician having ordinary knowledge in the art are within the scope of the present invention. For example, the structure (10) may be manufactured using the principle of nano-growth of metal particles in a solution, and may be manufactured by stirring a solution containing core-shell structured nanoparticles (11) and a solution containing TEOS and a biomolecule material. Preferably, the structure (10) may follow the method for manufacturing the structure and diagnostic method for surface-enhanced Raman scattering spectroscopy described above.

[0180] Referring again to FIG. 4a, the present diagnostic method may include a step (S200) of diagnosing a target disease by analyzing a second spectral signal acquired through the diagnostic solution (1) in the step (S100). At this time, as an optional embodiment, a step (S220) of diagnosing a target disease by comparing and analyzing the second spectral signal acquired from the diagnostic solution and spectral information (S210) may be included (see FIG. 4b).

[0181] As described above, biomolecular substances that appear specifically according to the specific disease may be distributed within the body of a patient suffering from a specific disease. According to one embodiment, a specific spectral signal can be acquired according to the biomolecular substance (B) within the structure (10), and by analyzing the signal, the target disease can be diagnosed. In one optional embodiment, the biomolecular substance (B) can be identified and the target disease can be diagnosed through this.

[0182] A diagnostic method according to one embodiment may include a step (not shown) of acquiring a first spectral signal from a biological solution extracted from a patient suffering from a specific disease or a solution containing the same. Additionally, as an optional example, the first spectral signal may be stored in a database, such as a data storage unit (140), to store spectral information.

[0183] At this time, the method for obtaining the first spectral signal is not particularly limited, and all surface-enhanced Raman scattering methods in the form of nanoparticles, substrates, liquids, or solids can be used. In addition, all methods for obtaining surface-enhanced Raman scattering spectral signals that can be easily selected by a technician with ordinary knowledge in the art should be interpreted as falling within the scope of the present invention.

[0184] In addition, as an optional embodiment, a step of acquiring a spectral signal for a biomolecular material (B) and storing the spectral information by storing the spectral signal for the biomolecular material (B) in a database (DB) may be included. At this time, the databased spectral information may be stored in a data storage unit (140) in an embodiment described later.

[0185] In another optional embodiment, the step of database-izing biomolecular substances (B) may include a step of database-izing biomolecular substances (B) that cause any disease by acquiring spectral signals using target bacteria and / or viruses that cause any disease.

[0186] Depending on the type of disease, specific biomolecules (B) may exist within the body. By analyzing these biomolecules from various perspectives, a method for diagnosing the target disease can be developed. As mentioned above, these biomolecules (B) could include not only proteins and sugars, but also bacteria and viruses.

[0187] Meanwhile, in one embodiment, referring again to FIG. 2B, the biomolecule material (B) may include two or more different types of biomolecule materials (B). Depending on the type of disease, biomolecules that specifically occur within the body may also include two or more types of biomolecule materials (B), and by specifying the biomolecule materials (B) in this way, the accuracy and reproducibility of the diagnosis of the target disease may be further improved.

[0188] In one embodiment, a method of comparing and analyzing the first spectral signal and the second spectral signal may use a method of comparing and confirming peak intensity, consistency, specific wave number, etc.

[0189] As an example, when comparing the first and second spectral signals, in the case of a pancreatic cancer patient, when obtaining the surface-enhanced Raman scattering signal, 1453 cm -1 , 878 cm -1 , 1517 cm -1 , 1385 cm -1 , 1303 cm -1 , 1153 cm -1 , 1184 cm -1 , 1043 cm -1 , 669 cm -1 , 435 cm -1 It may have a spectral signal with a peak at 1453 cm in the diagnostic solution as described above. -1 , 878 cm -1 , 1517 cm -1 , 1385 cm -1 , 1303 cm-1 , 1153 cm -1 , 1184 cm -1 , 1043 cm -1 , 669 cm -1 , 435 cm -1 If a spectral signal appears in the above-mentioned diagnostic solution, it can be diagnosed that the biomolecule substances contained in the diagnostic solution belong to a patient with pancreatic cancer.

[0190] In one embodiment, the diagnostic method may further include a step of comparing with a comparative experimental group. It can be confirmed whether the spectral signal obtained using the diagnostic solution (1) was obtained through the normal operation of the diagnostic system (100), and such comparative experimental group may be formed using the same solvent as the diagnostic solution (1).

[0191] Meanwhile, the steps for comparing with the comparative experimental group are not particularly limited, and any method that can be selected by a technician with common knowledge in the technical field should be interpreted as falling within the scope of the present invention.

[0192] FIGS. 5A to 5C are block diagrams showing a diagnostic system (100) provided by one embodiment.

[0193] In order to solve the above technical problem, another embodiment of the present invention provides a disease diagnosis system using surface-enhanced Raman scattering spectroscopy (hereinafter referred to as “the diagnosis system”) (100).

[0194] Referring to FIG. 5a, the diagnostic system (100) provided by one embodiment may include a container portion (110) in which a diagnostic solution (1) is placed, an irradiation portion (120) that irradiates light to the diagnostic solution (1) placed in the container portion (110) to obtain a spectral signal from one area of ​​the structure (10), and a diagnostic portion (130) that analyzes the obtained spectral signal.

[0195] In one embodiment, the diagnostic system (100) may further include a manufacturing unit for manufacturing a structure (10). The manufacturing unit may manufacture a structure (10) containing a biomolecule material (B) in one area of ​​the interior.

[0196] At this time, in an optional embodiment, the manufacturing unit can manufacture a diagnostic solution (1) in which the structure (10) is dispersed. In this way, the manufactured diagnostic solution (1) can be used in the container unit (110), the irradiation unit (120), etc., which will be described later.

[0197] Meanwhile, the method of manufacturing the structure (10) in the manufacturing department is not particularly limited, and a more specific and detailed description of the method of manufacturing the structure (10) is replaced with the description in the preceding embodiments.

[0198] In one embodiment, the diagnostic solution (1) may include a structure (10). The diagnostic system (100) can directly irradiate light to the diagnostic solution (1) through the irradiation unit (120) to obtain a spectral signal from an area of ​​the structure (10) placed in the diagnostic solution (1), and can diagnose a target disease by analyzing the spectral signal in the diagnostic unit (130).

[0199] Meanwhile, the structure (10) may include a biomolecule material (B) as described above, and a more specific and detailed description thereof will be replaced with the description in the preceding embodiments.

[0200] In one embodiment, the container part (110) may be configured to place a diagnostic solution (1), and light may be irradiated onto the diagnostic solution (1) placed in the container part (110) to generate a spectral signal in one area of ​​the structure (10). It is preferable that the diagnostic solution (1) be placed sufficiently, and as an optional example, the container part (110) in which the diagnostic solution (1) is placed may be configured to have a size of at least 200 μL, but it is to be understood that the present invention is not limited to this size.

[0201] Meanwhile, the container part (110) is not particularly limited, and can be easily selected by a technician with general knowledge in the art, and it should be interpreted that all container surfaces that allow liquid to be contained and generate a spectral signal fall within the scope of the present invention.

[0202] In one embodiment, the irradiation unit (120) is configured to irradiate light to the diagnostic solution (1). When light is irradiated to the diagnostic solution (1), a spectral signal is generated in one area within the structure (10) placed within the diagnostic solution (1).

[0203] As an optional example, the structure (10) may include nanoparticles (11), a biomolecular material (B), and a silica shell surrounding them. In addition, the spectral signal may be generated near the surface where the biomolecular material (B) and the nanoparticles (11) meet. Meanwhile, a more specific and detailed description of the structure (10) will be replaced with the description in the preceding embodiments.

[0204] Referring again to FIG. 5A, in one embodiment, the irradiation unit (120) may include an irradiation unit that emits light. The irradiation units are not particularly limited, and in one optional embodiment, may include a laser irradiation unit.

[0205] Referring to FIG. 5b, in one embodiment, the diagnostic unit (130) can diagnose a target disease by comparing and analyzing the spectral information from the aforementioned embodiments with the second spectral signals. As an optional embodiment, the biomolecule material (B) contained in the diagnostic solution (1) can be identified during this process.

[0206] Meanwhile, when using information on a spectral signal in the diagnostic unit (130), spectral information on the first spectral signal stored in the data storage unit (140) described later can be used.

[0207] In one embodiment, the diagnostic unit (130) may use an algorithm that compares and / or analyzes the peak frequency coincidence, peak intensity, coincidence, etc. between the spectral information and the second spectral signal, and through the above process, the first spectral signal can be searched for, and a target disease that specifically corresponds to the searched first spectral signal can be diagnosed.

[0208] Referring to FIG. 3c, in one embodiment, the diagnostic unit (130) may include a first diagnostic unit (130a) that searches for a first spectral signal corresponding to a second spectral signal among spectral information, and a second diagnostic unit (130b) that diagnoses a disease using the searched first spectral signal.

[0209] Referring again to FIG. 5b, in one embodiment, the diagnostic system (100) may include a data storage unit (140) that acquires a spectral signal and stores spectral information.

[0210] In one embodiment, the data storage unit (140) can store spectral information for the first spectral signal described above. The diagnosis unit (130) can use the spectral information stored in the data storage unit (140) to compare it with a second spectral signal, thereby searching for a first spectral signal corresponding to the second spectral signal among the spectral information. In addition, a specific disease can be searched for through the first spectral signal, and through this, the diagnosis unit (130) can diagnose the target disease.

[0211] Referring again to FIGS. 5b and 5c, in one embodiment, the data storage unit (140) may include a first data storage unit (140a) that stores spectral information for a first spectral signal. The spectral information may include, for example, a spectrum chart of a surface-enhanced Raman scattering spectral signal for a specific disease, spectral intensity, and irradiance setting values, but is not limited to the above examples.

[0212] In one embodiment, the data storage unit (140) may include a second data storage unit (140b) that stores information about a disease and the first spectral signal. The diagnosis unit (130) may diagnose the target disease using the information recorded in the second data storage unit (140b).

[0213] As an optional embodiment, the data storage unit (140) may store information on biomolecular substances (B) in a database. Such information on the spectral signal of the biomolecular substances (B) may be, for example, a specific Raman scattering spectral signal generated depending on the type of substance, such as a protein, bacteria, virus, or mineral. The description of the method for obtaining such information will be replaced with the description in the preceding embodiment.

[0214] Meanwhile, in one embodiment, the data storage unit (140) and the diagnostic unit (130) may communicate using a wired / wireless communication method, and a diagnostic system (100) for diagnosing a disease through near / far distance methods may be provided through such a method. Meanwhile, such wired / wireless communication methods are not particularly limited, and any communication method that can be easily selected by a technician with ordinary knowledge in the art should be interpreted as falling within the scope of the present invention.

[0215] It goes without saying that the embodiments of the present invention described above can be applied in combination with each other.

[0216]

[0217] Hereinafter, manufacturing examples, examples, and experimental examples of the present invention will be described. However, it should be noted that these manufacturing examples, examples, and experimental examples are intended to more specifically explain the composition and effects of the present invention and are not intended to limit the scope of the present invention.

[0218]

[0219] Example

[0220] Example 1 - Structure comprising nanoparticles having a core-shell structure

[0221] In this Example 1, a surface-enhanced Raman scattering spectroscopy structure and a surface-enhanced Raman scattering spectroscopy composition in which the structure is dispersed were manufactured. The specific manufacturing process is as follows.

[0222] In order to manufacture a surface-enhanced Raman scattering spectroscopy structure and a surface-enhanced Raman scattering spectroscopy composition in which the structure is dispersed, nanoparticles were formed, and then a biomolecular material located near the formed nanoparticles and a silica shell layer surrounding them were formed together.

[0223] (1) Formation of core-shell nanoparticles

[0224] To form 2-3 nm sized gold nanoparticles capped with CTAB, 9.75 ml of 100 mM CTAB solution was prepared, followed by 10 mM HAuCl 4· 250 μl of 3H2O solution and 600 μl of 10 mM NaBH4 solution were added, stirred for 2 minutes, and the reaction was allowed to proceed for 3 hours.

[0225] Afterwards, in order to grow 30 nm gold nanoparticles using the 2 to 3 nm sized gold nanoparticles capped with the above-formed CTAB, 16 mL of 200 mM CTAC solution was prepared, followed by 0.5 mM HAuCl 4· 16 mL of 3H2O solution and 16 mL of 0.8 mM ascorbic acid solution were added to 40 μL of previously prepared CTAB-capped AuNP seeds, and the reaction was carried out for 7 days.

[0226] Thereafter, to form a second plasmonic metal shell layer of silver on the surface of the prepared 30 nm gold nanoparticles to form core-shell nanoparticles, 10 mL of a 1.0 mM AgNO3 solution and 10 mL of a 25 mM ascorbic acid solution were added to 10 mL of the formed 30 nm AuNP solution, and the reaction was allowed to proceed for 40 minutes. Thereafter, after centrifugation, the solution was redispersed in triple-distilled water to form gold-silver core-shell nanoparticles.

[0227] (2) PVP coating on the surface of core-shell structure nanoparticles

[0228] After preparing 10 mL of the previously formed Au-Ag core-shell nanoparticle solution, 0.5 mL of 1 v / v % SDS solution and 5 mL of 2.5 % (w / v) PVP solution were added, reacted for 10 minutes, centrifuged, and redispersed in distilled water, thereby forming core-shell nanoparticles with a PVP coating layer formed on the surface.

[0229] (3) Formation of biomolecule material and silica shell layer

[0230] In order to form core-shell nanoparticles having a previously formed PVP coating layer, biomolecule materials located near the nanoparticles, and a silica shell layer surrounding them, the process was carried out in the following order. In this case, in Example 1.1, a surface-enhanced Raman scattering spectroscopy structure was manufactured after pretreating plasma with ethanol, and in Example 1.2 below, a surface-enhanced Raman scattering spectroscopy structure was manufactured using plasma that had not undergone a pretreatment process.

[0231]

[0232] Example 1.1 - Preprocessing in progress

[0233] After preparing 10 μL of plasma, a pretreatment process was performed on the plasma by adding 20 μL of deionized water and 70 μL of ethanol (EtOH) and allowing the reaction to proceed for 10 minutes.

[0234] 17.5 μL of core-shell nanoparticles having a PVP coating layer formed thereon, 20 μL of ethanol-pretreated plasma supernatant, 44.5 μL of deionized water, and 20 μL of 0.05 v / v % TEOS solution were added, and the reaction was allowed to proceed for 30 minutes. Then, 20 μL of 10 v / v % DMA solution was added thereto, to prepare a composition for surface-enhanced Raman scattering spectroscopy in which a structure for surface-enhanced Raman scattering spectroscopy was dispersed.

[0235] Example 1.2 - No preprocessing

[0236] 17.5 μL of core-shell nanoparticles having a PVP coating layer formed thereon, 2 μL of plasma not pretreated with ethanol, 60.5 μL of deionized water, and 20 μL of a 0.05 v / v % TEOS solution were added, and the reaction was allowed to proceed for 30 minutes. Then, 20 μL of a 10 v / v % DMA solution was added thereto to prepare a composition for surface-enhanced Raman scattering spectroscopy in which a structure for surface-enhanced Raman scattering spectroscopy was dispersed.

[0237] Experimental Example 1 - Structure Verification

[0238] In this Experimental Example 1, an experiment was conducted to confirm the state of the surface-enhanced Raman scattering spectroscopy structure manufactured through the above Examples 1.1 and 1.2.

[0239]

[0240] Experimental Example 1.1 - Determining the Size of Nanoparticles

[0241] In this Experimental Example 1.1, an experiment was conducted to determine the size of nanoparticles. The specific experimental method involved measuring the size of nanoparticles using dynamic light scattering and TEM imaging.

[0242] Nanoparticle size measurements using dynamic light scattering revealed that the spherical gold nanocore had a diameter of 31 nm and a PI of 0.16. The cubic gold core-shell nanoparticles had a diameter of 47.5 nm and a PI of 0.25.

[0243] Referring to the TEM image of Fig. 6a, the width of the gold core-shell nanoparticles was measured to be approximately 30 nm, and it was confirmed that a spherical gold core existed inside the cubic silver metal shell layer.

[0244]

[0245] Experimental Example 1.2 - Confirmation of the silica shell layer

[0246] In this Experimental Example 1.2, an experiment was conducted to determine whether a silica shell layer was formed on the surface of the structure. The specific experimental method involved measuring nanoparticle size using dynamic light scattering and TEM imaging.

[0247] The diameter of the nanoparticles formed with a silica shell layer using dynamic light scattering is 90.25 nm and the PI is 0.35 when pretreated with ethanol, and 87.4 nm and the PI is 0.31 when not pretreated with ethanol. The thickness of the silica shell is 23.28 nm when pretreated with ethanol, and 21.85 nm when not pretreated.

[0248] Referring to the TEM image of Fig. 6b, it was confirmed that a silica shell layer was formed outside the gold-silver core-shell structure, which consists of a gold core located in the center and a cubic silver metal shell layer surrounding the gold core.

[0249]

[0250] Experimental Example 1.3 - Identification of Biomolecular Substances

[0251] In this experimental example 1.3, the Raman scattering spectra were compared when a biomaterial was present and when it was not present inside the structure.

[0252] The specific experimental method confirmed the difference in Raman spectra between the silica shell formed by adding a plasma sample to PVP-coated silver (Ag) nanocube particles (red) and the silica shell formed without adding a plasma sample (blue).

[0253] Figure 7 is a drawing showing the experimental results of Experimental Example 1.3.

[0254] Referring to Figure 7, when examining the difference in Raman spectra when a plasma sample, i.e. a biomaterial, is present and absent within the nanostructure, it can be seen that a number of peaks are newly created or disappear in the spectrum. For example, 1170 cm -1 , 1520 cm -1 The peak of the representative is newly created as new plasma exists, 930 cm -1 , 1130cm -1 The peak of the representative disappears.

[0255]

[0256] Experimental Example 2 - Raman Spectrum Verification Experiment with and without Pretreatment

[0257] In this experimental example 2, an experiment was conducted to confirm what difference occurs in the Raman spectrum depending on whether ethanol was pretreated or not, through the above examples 1.1 and 1.2.

[0258] As a specific experimental method, the process of stacking a silica shell with plasma and nanoparticles was carried out, 70% ethanol and plasma were mixed in a ratio of 9:1 (v / v) and left for 10 minutes, and then the process of stacking a silica shell with nanoparticles was carried out, and the Raman spectra were obtained and compared.

[0259] Figure 8 is a Raman spectrum showing the experimental results of Experimental Example 2.

[0260] Referring to Figure 8, comparing the case without pretreatment (blue) and the case with ethanol pretreatment (orange), the case with pretreatment is 650 cm -1 , 880cm -1 A peak exists at 1280 cm when no pretreatment is performed. -1 , 1390 cm -1 It can be seen that there is a peak.

[0261]

[0262] Experimental Example 3 - Confirmation of Surface-Enhanced Raman Scattering Spectroscopic Signals for Diagnosis of Target Diseases

[0263] In this experimental example 3, a surface-enhanced Raman scattering spectroscopy signal was obtained using the diagnostic solution manufactured through the above example 1.

[0264] The diagnostic solution was placed in a 96-well plate, irradiated with a 532 nm laser, and the scattered light was acquired using a Raman spectrometer. The results were as shown in Figure 9.

[0265]

[0266] Experimental Example 4 - Confirming the Diagnosis Error Rate of the Target Disease

[0267] In this experimental example 4, the target disease was diagnosed using the spectral signal obtained through the experimental example 3 above. 1453 cm -1 , 878 cm -1 , 1517 cm -1 , 1385 cm -1 , 1303 cm -1 , 1153 cm -1 , 1184 cm -1 , 1043 cm -1 , 669 cm -1 , 435 cm -1 As a result of confirming the peaks that appeared in , normal people and cancer patients were classified with a precision of 0.931, a sensitivity of 0.800, and a specificity of 0.800.

[0268] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics 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 entity may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined manner.

[0269] The scope of the present invention is indicated by the claims described below, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. A silica shell layer having an internal storage space; and Nanoparticles comprising a plasmonic metal; A surface-enhanced Raman scattering spectroscopy structure in which the above nanoparticles are arranged in one area of ​​the receiving space inside the silica shell layer.

2. In paragraph 1, The nanoparticle comprises a core comprising a first plasmonic metal and a second plasmonic metal shell layer arranged to surround the core and comprising a second plasmonic metal, A surface-enhanced Raman scattering spectroscopy structure in which the second plasmonic metal shell layer has a symmetrical structure based on the center of the second plasmonic metal shell layer.

3. In paragraph 1, The above structure is a surface-enhanced Raman scattering spectroscopy structure including a biomolecular material arranged in the receiving space.

4. In paragraph 2, A surface-enhanced Raman scattering spectroscopy structure, wherein the first plasmonic metal and the second plasmonic metal each independently include at least one selected from the group consisting of Au, Ag, Cu, Al, W, Pt, Ni, and Pd.

5. In paragraph 2, A surface-enhanced Raman scattering spectroscopy structure, wherein the width of the plasmon resonance peak on the surface of the second plasmonic metal is narrower than the width of the plasmon resonance peak on the surface of the first plasmonic metal.

6. In paragraph 2, The above second plasmonic metal shell layer is a surface-enhanced Raman scattering spectroscopy structure having a regular polyhedral structure.

7. A step of preparing a solution in which nanoparticles are dispersed; and A step of forming a silica shell layer surrounding nanoparticles dispersed in the above solution; A method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, comprising:

8. In paragraph 7, The step of preparing a solution in which the above nanoparticles are dispersed includes a step of forming a mixed solution by mixing a first solution containing a first precursor compound including a first plasmonic metal and a second solution containing a second precursor compound including a second plasmonic metal, The nanoparticle comprises a core comprising a first plasmonic metal and a second plasmonic metal shell layer arranged to surround the core and comprising a second plasmonic metal, A method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, wherein the second plasmonic metal shell layer has a symmetrical structure based on the center of the second plasmonic metal shell layer.

9. In paragraph 8, A method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, further comprising a step of mixing a coating solution containing a water-soluble polymer compound into the above mixed solution.

10. In paragraph 8, The step of forming the above silica shell layer includes a step of mixing the mixed solution, a third solution containing a biomolecule material, and a fourth solution containing a silica precursor compound; A method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, wherein the silica shell layer is formed so that at least some of the nanoparticles and the biomolecule material are arranged together inside.

11. A step of obtaining a first surface-enhanced Raman scattering spectroscopy signal from a biological solution extracted from a patient with a disease; A step of irradiating light on a diagnostic solution in which a surface-enhanced Raman scattering spectroscopy structure according to any one of claims 1 to 6 is dispersed to obtain a second surface-enhanced Raman scattering spectroscopy signal in one region of the surface-enhanced Raman scattering spectroscopy structure; and A step of identifying the disease of the patient by comparing the first surface-enhanced Raman scattering spectroscopy signal and the second surface-enhanced Raman scattering spectroscopy signal; A disease diagnosis method using surface-enhanced Raman scattering spectroscopy, including:

12. In paragraph 11, The above surface-enhanced Raman scattering spectroscopy structure is a disease diagnosis method using surface-enhanced Raman scattering spectroscopy, which includes two or more types of biomolecular substances.

13. In paragraph 11, The above diagnostic method comprises the steps of: creating a database of the first surface-enhanced Raman scattering spectroscopy signal obtained from a biological solution extracted from the patient; A disease diagnosis method using surface-enhanced Raman scattering spectroscopy, further comprising:

14. In paragraph 11, The above diagnostic method comprises the steps of: obtaining a surface-enhanced Raman scattering spectroscopy signal for a biomolecular substance within the surface-enhanced Raman scattering spectroscopy structure, and creating a database of information on the surface-enhanced Raman scattering spectroscopy signal for the biomolecular substance; A disease diagnosis method using surface-enhanced Raman scattering spectroscopy, further comprising:

15. Container section where the diagnostic solution is placed; An irradiation unit that irradiates light to a diagnostic solution placed in the above-mentioned container and generates a surface-enhanced Raman scattering spectroscopy signal in one area of ​​a surface-enhanced Raman scattering spectroscopy structure according to any one of claims 1 to 6 included in the diagnostic solution; and A diagnostic unit that analyzes the surface-enhanced Raman scattering spectroscopy signal generated above; A disease diagnosis system using surface-enhanced Raman scattering spectroscopy, including:

16. In paragraph 15, A disease diagnosis system using surface-enhanced Raman scattering spectroscopy, wherein the surface-enhanced Raman scattering spectroscopy structure comprises two or more types of biomolecular substances.

17. In paragraph 15, The above diagnostic system comprises a data storage unit that acquires the surface-enhanced Raman scattering spectroscopy signal and stores spectral information for the surface-enhanced Raman scattering spectroscopy signal; A disease diagnosis system using surface-enhanced Raman scattering spectroscopy, which further includes:

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