Structure for surface-enhanced raman scattering spectrometry, method for manufacturing same, and

Abstract

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. According to the structure for surface-enhanced Raman scattering spectroscopy, the method for manufacturing the same, and the diagnostic method using the same of the present invention, it is possible to provide a structure for surface-enhanced Raman scattering spectroscopy, which can be used as a surface-enhanced Raman scattering spectroscopy, even if there is no biomarker having excellent diagnostic performance in blood indexes. A structure for surface-enhanced Raman scattering spectroscopy, which can be used even for intractable cancers such as pancreatic cancer and intractable diseases, and a method for producing the same. Further, a diagnostic method and a diagnostic system using a surface-enhanced Raman scattering spectrum can be provided by the structure for the surface-enhanced Raman scattering spectrum.

Description

Technical Field

[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. Background Art

[0002] Surface - enhanced Raman scattering (SERS) spectroscopy is a spectroscopic method developed to compensate for the weak signal and poor reproducibility of Raman scattering spectroscopy. It is a spectroscopic method that utilizes the phenomenon that the Raman scattering intensity of molecules adsorbed on the surface of metal nanostructures such as gold and silver increases rapidly by more than 10^6 to 10^8 times.

[0003] Surface - enhanced Raman scattering (SERS) spectroscopy is a technique that can obtain a large amount of information through a single measurement, with ultra - sensitivity sufficient to directly measure single molecules, and can directly measure information on the vibrational state or molecular structure of molecules. Therefore, it is recognized as a powerful analytical method for chemical / biological / biochemical analysis.

[0004] In recent years, surface - enhanced Raman scattering spectroscopy has attracted much attention as a method for accurately obtaining comprehensive information on biomolecules from biological analysis samples. However, the previously proposed methods for detecting biomolecules using surface - enhanced Raman scattering spectroscopy require specific binding, and there are no biomarkers with excellent diagnostic performance in the blood indicators of intractable cancers such as pancreatic cancer and intractable diseases. Therefore, there are problems in screening and early diagnosis.

[0005] Therefore, there is a need for a diagnostic method and diagnostic system using surface - enhanced Raman scattering spectroscopy, which can be used even in the case where there are no biomarkers with excellent diagnostic performance in biological indicators, such as intractable cancers like pancreatic cancer and intractable diseases.

[0006] On the other hand, the content described as background art is only used to enhance the understanding of the background of the present invention and should not be regarded as an admission that they correspond to the prior art known to those of ordinary skill in the art. Summary of the Invention

[0007] Technical Problem

[0008] The technical problem to be solved by the present invention is to provide a structure for surface - enhanced Raman scattering spectroscopy and a method for manufacturing the same, which can be used even in the case where there are no biomarkers with excellent diagnostic performance in blood indicators, such as intractable cancers like pancreatic cancer and intractable diseases.

[0009] The technical problem to be solved by the present invention is to provide a disease diagnostic method and diagnostic system using surface - enhanced Raman scattering spectroscopy, which can be used even in the case where there are no biomarkers with excellent diagnostic performance in biological indicators.

[0010] In addition, the technical problem to be solved by the present invention is to provide a diagnostic method and a diagnostic system using surface enhanced Raman scattering spectroscopy with high reproducibility and convenient for circulation and storage.

[0011] The technical problems to be solved by the present invention are not limited to the above technical problems. For those of ordinary skill in the technical field to which the present invention pertains, other technical problems not mentioned can be clearly understood from the following description.

[0012] Technical solution

[0013] To solve the above technical problems, an embodiment of the present invention provides a structure for surface enhanced Raman scattering spectroscopy, which includes: a silica shell having an accommodation space inside; and nanoparticles having a plasmonic metal, wherein the nanoparticles are distributed in a region of the accommodation space inside the silica shell.

[0014] To solve the above technical problems, another embodiment of the present invention provides a method for manufacturing a structure for surface enhanced Raman scattering spectroscopy, which includes the following steps: preparing a solution in which nanoparticles are dispersed; and forming a silica shell surrounding the nanoparticles dispersed in the solution.

[0015] To solve the above technical problems, an embodiment of the present invention provides a disease diagnosis method using surface enhanced Raman scattering spectroscopy, which includes the following steps: obtaining a first surface enhanced Raman scattering spectrum signal from a biological solution extracted from a patient; irradiating light onto a diagnostic solution in which metal nanostructures are dispersed to obtain a second surface enhanced Raman scattering spectrum signal in a region of the metal nanostructures, wherein a region inside the metal nanostructure has a biomolecular material; and comparing the first surface enhanced Raman scattering spectrum signal with the second surface enhanced Raman scattering spectrum signal to identify the disease of the patient.

[0016] To solve the above technical problems, another embodiment of the present invention provides a disease diagnosis system using surface enhanced Raman scattering spectroscopy, which includes: a container part for placing a diagnostic solution; an irradiation part for irradiating light onto the diagnostic solution placed in the container part and generating a surface enhanced Raman scattering spectrum signal in a region of the metal nanostructures contained in the diagnostic solution; and a diagnosis part for analyzing the generated surface enhanced Raman scattering spectrum signal.

[0017] Beneficial effects

[0018] According to an embodiment of the present invention, a structure for surface-enhanced Raman scattering spectroscopy and a manufacturing method thereof can be provided, which can non-specifically provide surface-enhanced Raman scattering spectral signals with biomolecular materials.

[0019] In addition, according to an embodiment of the present invention, a structure for surface-enhanced Raman scattering spectroscopy and a manufacturing method thereof can be provided, which can be used even in cases where there are no biomarkers with excellent diagnostic performance in blood indicators, such as refractory cancers and refractory diseases like pancreatic cancer.

[0020] According to an embodiment of the present invention, a diagnostic method and a diagnostic system using surface-enhanced Raman scattering spectroscopy can be provided, which can be used even in cases where there are no biomarkers with excellent diagnostic performance in biological indicators.

[0021] In addition, according to an embodiment of the present invention, a diagnostic method and a diagnostic system using surface-enhanced Raman scattering spectroscopy with high repeatability and convenient for circulation and preservation can be provided.

[0022] The effects of the present invention are not limited to the above effects, but should be understood to include all effects that can be inferred from the composition of the present invention described in the detailed description or claims of the present invention.

[0023] Brief Description of the Drawings

[0024] Figures 1a to 1e It is a conceptual diagram of a structure for surface-enhanced Raman scattering spectroscopy provided according to an embodiment of the present invention.

[0025] Figure 2a and Figure 2b It is a conceptual diagram of a diagnostic solution for surface-enhanced Raman scattering spectroscopy provided according to an embodiment.

[0026] Figure 3a and Figure 3b It is a flowchart of a manufacturing method of a structure for surface-enhanced Raman scattering spectroscopy provided according to an embodiment.

[0027] Figure 4a and Figure 4b It is a flowchart of a diagnostic method using surface-enhanced Raman scattering spectroscopy provided according to an embodiment.

[0028] Figures 5a to 5c It is a block diagram showing a diagnostic system provided according to an embodiment.

[0029] Figure 6a and Figure 6b It is a transmission electron microscope (TEM) image of a structure for surface-enhanced Raman scattering spectroscopy provided according to an embodiment.

[0030] Figure 7 A graph showing the results of Raman scattering experiments when a biological material is present and when it is absent inside a structure according to an embodiment.

[0031] Figure 8 A graph showing the results of experiments on Raman spectral differences according to whether ethanol pretreatment is performed or not according to an embodiment.

[0032] Figure 9 A graph for verifying the surface-enhanced Raman scattering spectral signal of a diagnostic solution provided according to an embodiment.

[0033] <Symbol Explanation>

[0034] 1: Diagnostic solution

[0035] 10: Structure

[0036] 11: Nanoparticle

[0037] 12: Silica shell

[0038] 20: Dispersion

[0039] 100: Diagnostic system

[0040] 110: Container part

[0041] 111: Core

[0042] 112: Second plasmonic metal shell

[0043] 113: Polymer coating

[0044] 120: Irradiation part

[0045] 130: Diagnostic part

[0046] 140: Data storage part

[0047] Best Mode

[0048] An embodiment of the present invention can provide a structure for surface-enhanced Raman scattering spectroscopy, which includes: a silica shell having an accommodation space inside; and nanoparticles having plasmonic metal, wherein the nanoparticles are distributed in a region of the accommodation space inside the silica shell.

[0049] An embodiment of the present invention can provide a structure for surface-enhanced Raman scattering spectroscopy, wherein the nanoparticle includes: a core including a first plasmonic metal; and a second plasmonic metal shell surrounding and distributed around the core and having a second plasmonic metal, and the second plasmonic metal shell has a symmetric structure with respect to the center of the second plasmonic metal shell.

[0050] In addition, in an embodiment of the present invention, the structure can be a structure for surface-enhanced Raman scattering spectroscopy, which includes a biomolecular material placed in the accommodation space.

[0051] In addition, an embodiment of the present invention can be a structure for surface-enhanced Raman scattering spectroscopy, wherein the first plasmonic metal and the second plasmonic metal each independently include any one or more selected from the group consisting of Au, Ag, Cu, Al, W, Pt, Ni, and Pd.

[0052] In addition, an embodiment of the present invention can be a structure for surface-enhanced Raman scattering spectroscopy, 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.

[0053] In addition, an embodiment of the present invention can be a structure for surface-enhanced Raman scattering spectroscopy, wherein the second plasmonic metal shell can have a regular polyhedron structure.

[0054] Another embodiment of the present invention provides a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, which includes the following steps: preparing a solution in which nanoparticles are dispersed; and forming a silica shell surrounding the nanoparticles dispersed in the solution.

[0055] An 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 nanoparticles are dispersed includes: mixing a first solution containing a first precursor compound with a second solution containing a second precursor compound to form a mixed solution, the first precursor compound contains a first plasmonic metal, the second precursor compound contains a second plasmonic metal, wherein the nanoparticle includes a core and a second plasmonic metal shell, the core contains a first plasmonic metal, the second plasmonic metal shell surrounds and is distributed around the core and has a second plasmonic metal, and the second plasmonic metal shell has a symmetric structure with respect to the center of the second plasmonic metal shell.

[0056] In addition, an embodiment of the present invention may be a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, which further includes a step of mixing a coating solution containing a water-soluble polymer compound into the mixed solution.

[0057] In addition, an embodiment of the present invention may provide a method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, wherein the step of forming the silica shell layer includes: mixing the mixed solution with a third solution containing a biomolecular material and a fourth solution containing a silica precursor compound, and the silica shell layer allows at least a part of the nanoparticles and the biomolecular material to be distributed together inside it.

[0058] Another embodiment of the present invention may provide a method for diagnosing a disease using surface-enhanced Raman scattering spectroscopy, which includes the following steps: obtaining a first surface-enhanced Raman scattering spectrum signal from a biological solution extracted from a patient; irradiating light onto a diagnostic solution in which a structure for surface-enhanced Raman scattering spectroscopy is dispersed to obtain a second surface-enhanced Raman scattering spectrum signal in a region of the structure for surface-enhanced Raman scattering spectroscopy, wherein a region inside the structure for surface-enhanced Raman scattering spectroscopy has a biomolecular material; and comparing the first surface-enhanced Raman scattering spectrum signal with the second surface-enhanced Raman scattering spectrum signal to identify the disease of the patient.

[0059] In addition, an embodiment of the present invention may be a method for diagnosing a disease using surface-enhanced Raman scattering spectroscopy, wherein the structure for surface-enhanced Raman scattering spectroscopy further includes: a silica shell layer having an accommodation space inside; and nanoparticles having a plasmonic metal, wherein the nanoparticles and the biomolecular material are distributed in the accommodation space inside the silica shell layer.

[0060] In addition, an embodiment of the present invention may be a method for diagnosing a disease using surface-enhanced Raman scattering spectroscopy, wherein the biomolecular material includes two or more types of biomolecular materials.

[0061] In addition, an embodiment of the present invention may be a method for diagnosing a disease using surface-enhanced Raman scattering spectroscopy, wherein the diagnostic method further includes the following step: establishing a database of the first surface-enhanced Raman scattering spectrum signals obtained from the biological solution extracted from the patient.

[0062] In addition, an embodiment of the present invention may be a disease diagnosis method using surface-enhanced Raman scattering spectroscopy. In this method, the diagnosis method further includes the following steps: obtaining the surface-enhanced Raman scattering spectrum signal of the biomolecular material to establish a database of the surface-enhanced Raman scattering spectrum signal information of the biomolecular material.

[0063] Another embodiment of the present invention may provide a disease diagnosis system using surface-enhanced Raman scattering spectroscopy, which includes: a container part for placing a diagnostic solution; an irradiation part for irradiating light on the diagnostic solution placed in the container part and generating a surface-enhanced Raman scattering spectrum signal in a region of the metal nanostructure contained in the diagnostic solution; and a diagnosis part for analyzing the generated surface-enhanced Raman scattering spectrum signal.

[0064] An embodiment may be a disease diagnosis system using surface-enhanced Raman scattering spectroscopy. In this system, the structure for surface-enhanced Raman scattering spectroscopy further includes: a silica shell layer having an accommodation space inside; and nanoparticles having a plasmonic metal, wherein the nanoparticles and the biomolecular material are distributed in the accommodation space inside the silica shell layer.

[0065] In addition, an embodiment may be a disease diagnosis system using surface-enhanced Raman scattering spectroscopy, wherein the biomolecular material includes two or more types of biomolecular materials.

[0066] In addition, an embodiment may be a disease diagnosis system using surface-enhanced Raman scattering spectroscopy. The diagnosis system may further include a data storage part for obtaining the surface-enhanced Raman scattering spectrum signal and storing the spectral information for the surface-enhanced Raman scattering spectrum signal. Detailed Embodiments

[0067] The present invention will be described below with reference to the accompanying drawings. However, the present invention can be implemented in various different forms and is not limited to the embodiments described herein. And, in order to clearly describe the present invention in the drawings, parts not relevant to the description are omitted, and similar parts are denoted by similar reference numerals throughout the specification.

[0068] In the drawings, for convenience of description, the dimensions of the constituent elements may be exaggerated or reduced. For example, the dimensions and thicknesses of the respective constituent elements shown in the drawings may be arbitrarily shown for convenience of description, and the following examples are not necessarily limited to what is shown in the drawings.

[0069] Terms such as first, second, A, B, etc. may be used to describe various components, but the components should not be limited by such terms. The terms are only used to distinguish one component from another. For example, without departing from the scope of the claims of the present invention, the first component may be named the second component, and similarly, the second component may also be named the first component. The term "and / or" includes any combination of a plurality of related described items or any item among a plurality of related described items.

[0070] Throughout the specification, when it is said that a part is "connected (joined, contacted, coupled)" to another part, this includes not only the case of "direct connection" but also the case of "indirect connection" with another part in between.

[0071] In addition, when a part is said to "include / contain / have" a certain component, this does not mean that it excludes other components, but it may also include other components, unless otherwise clearly stated.

[0072] The terms used herein are only for describing specific embodiments and are not intended to limit the present invention. Unless the context clearly indicates otherwise, singular expressions include plural expressions.

[0073] In this specification, it should be understood that terms such as "include / contain / have" are intended to specify the existence of the features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, but do not preclude the possibility of the existence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0074] When something is said to be "on" or "on another object", it includes not only the case where the object is directly above another object but also the case where the object is located in between.

[0075] Previously proposed methods for detecting biomolecules using surface-enhanced Raman scattering spectroscopy utilize the specific binding that occurs between biomolecules and metal nanoparticles and form surface-enhanced Raman scattering spectral signals at that position. Therefore, even when using surface-enhanced Raman scattering spectroscopy, there are problems such as difficulty in applying surface-enhanced Raman scattering spectroscopy to diseases such as pancreatic cancer that do not have excellent diagnostic biomarkers.

[0076] Figures 1a to 1e It is a conceptual diagram of a structure for surface-enhanced Raman scattering spectroscopy provided according to an embodiment of the present invention.

[0077] See Figure 1a, To solve the above technical problems, an embodiment of the present invention provides a structure 10 for surface-enhanced Raman scattering spectroscopy (hereinafter referred to as "structure") and a composition for surface-enhanced Raman scattering spectroscopy in which the structure 10 is dispersed, and a manufacturing method thereof. Among them, the structure 10 for surface-enhanced Raman scattering spectroscopy includes nanoparticles 11 and a silica shell layer 12.

[0078] In addition, as an optional embodiment, the interior of the silica shell layer 12 may include a biomolecular material B described later. Through this structure, without excluding a specific biomolecular material B, surface-enhanced Raman scattering spectral signals of a large amount of biomolecular material B non-specifically distributed inside the structure 10 can be obtained.

[0079] In addition, by using the structure 10 as described above and a composition for surface-enhanced Raman scattering spectroscopy in which the structure 10 is dispersed, a database of surface-enhanced Raman scattering signals is established, and a diagnostic kit with excellent diagnostic performance can be provided even for diseases without biomarkers with excellent spectral properties.

[0080] Hereinafter, the structure 10 will be described.

[0081] In one embodiment, the nanoparticles 11 may include a plasmonic metal. When a molecule or particle is present near the surface of the plasmonic metal, the Raman scattering signal can be increased through the local surface plasmon phenomenon, thereby obtaining a surface-enhanced Raman scattering spectral signal.

[0082] In this specification, the term "plasmonic metal" may refer to a metal excited in a magnetic field, and examples thereof include Au, Ag, Cu, Al, W, Pt, Ni, and Pd, etc.

[0083] In one embodiment, the nanoparticles 11 may include a core 111 and a second plasmonic metal shell layer 112. The core 111 includes a first plasmonic metal, and the second plasmonic metal shell layer 112 is distributed to surround the core 111 and includes a second plasmonic metal. By making the nanoparticles 11 have a core-shell structure as described above, nanoparticles 11 having the properties of both the first plasmonic metal and the second plasmonic metal can be provided.

[0084] In one embodiment, the second plasmonic metal shell layer 112 may have a symmetric structure based on the center M of the second plasmonic metal shell layer 112. By making the second plasmonic metal shell layer 112 surround the core 111 and have a symmetric structure as described above, the surface area between the nanoparticles 11 and the biomolecular material B can be increased, thereby maximizing the surface-enhanced Raman scattering spectral signal.

[0085] In one embodiment, the second plasma metal shell layer 112 may have a regular polyhedron structure, and the regular polyhedron may include at least one of, for example, a regular hexahedron, a regular octahedron, a regular dodecahedron, and a regular icosahedron.

[0086] In one embodiment, the core 111 includes a first plasma metal, and any metal that can provide a surface-enhanced Raman scattering spectrum signal through plasma phenomena on the surface should be understood to fall within the scope of the claims of the present invention. For example, Au, Ag, Cu, Al, W, Pt, Ni, and Pd, but not limited to the above examples.

[0087] In one embodiment, the core 111 may have an average particle size D of 50 nm or less. 50 . As another example, it may be 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 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.

[0088] When having the average particle size and the structure 10 is distributed in the dispersion liquid 20 described later, excellent Raman scattering effects can be provided.

[0089] In one embodiment, the core 111 may have a diameter of 10 nm to 50 nm in at least one direction. By having such a structure, when the structure 10 is distributed in the dispersion liquid 20 described later, excellent Raman scattering effects can be provided.

[0090] In one embodiment, the second plasma metal shell layer 112 includes a second plasma metal, and the width of the plasma resonance peak on the surface of the second plasma metal may be narrower than the width of the plasma resonance peak on the surface of the first plasma metal. With such a structure, by making the width of the plasma resonance peak on the surface of the second plasma metal narrower, better sensitivity can be provided when obtaining Raman scattering signals when the structure 10 is dispersed in the dispersion liquid 20 described later.

[0091] At this time, as an optional embodiment, the widths of the plasma resonance peaks of the first plasma metal surface and the second plasma metal surface can be measured in the same environment.

[0092] In one embodiment, the second plasma metal shell 112 includes a second plasma metal, which may include, for example, Au, Ag, Cu, Al, W, Pt, Ni, and Pd, but is not limited to the above examples.

[0093] In one embodiment, the core 111 may include gold (Au), and the second plasma metal shell 112 may include silver (Ag). The light-concentrating effect of silver (Ag) is more than five times stronger than that of gold (Au), and the structure 10 including the nanoparticles 11 having the said structure can provide an excellent surface-enhanced Raman scattering spectral signal.

[0094] In one embodiment, the thickness of the second plasma metal shell 112 in at least one direction may be 10 nm to 70 nm. As another example, the thickness of the second plasma metal shell 112 may be 30 nm to 70 nm, 40 nm to 70 nm, 50 nm to 70 nm, 60 nm to 70 nm, 30 nm to 60 nm, 40 nm to 60 nm, or 50 nm to 60 nm. When the thickness of the second plasma metal shell 112 exceeds 70 nm, the size of the nanoparticles 11 distributed inside the structure 10 becomes too large, and the reproducibility of surface-enhanced Raman scattering may be reduced. When it is less than 30 nm, the gap between the shell and the core 111 becomes too narrow, which may reduce the reproducibility of surface-enhanced Raman scattering.

[0095] See Figure 1b , in one embodiment, the nanoparticles 11 may include a polymer coating 113, which is formed on the outer side of the second plasma metal shell 112 and surrounds the second plasma metal shell 112. The polymer coating 113 imparts a negative charge to the surface of the nanoparticles 11 and can prevent the nanoparticles 11 from aggregating together.

[0096] By having the said structure, during the manufacturing process of the structure 10, it is possible to prevent the nanoparticles 11 from aggregating together in the stage before the silica shell 12 is formed, and further, it is easier to form the silica shell 12.

[0097] In one embodiment, cetyltrimethylammonium chloride (CTAC), which is a surfactant, and polyvinylpyrrolidone (PVP), which is a polymer, can be coated on the surface of the metal nanoparticles 11. Cetyltrimethylammonium chloride (CTAC) can fix charged molecules on the surface of the nanoparticles 11 through electrostatic attraction, while polyvinylpyrrolidone (PVP) can fix various biomolecules on the surface through van der Waals forces and chelation. This is because during the formation of the silica shell layer, cetyltrimethylammonium chloride (CTAC) and polyvinylpyrrolidone (PVP) coated on the surface of the metal nanoparticles 11 fix the biomolecules in the blood on the nanoparticle surface, so that the biomolecules in the blood can be easily located in the accommodation space within the silica shell layer.

[0098] In one embodiment, the polymer coating 113 is not particularly limited, and any polymer coating 113 that can be easily selected by those skilled in the art with ordinary knowledge in this technical field should be understood to fall within the scope of the claims of the present invention.

[0099] On the other hand, the polymer coating 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.

[0100] See Figure 1c and Figure 1d , in one embodiment, the structure 10 may further include a biomolecular material B placed in the accommodation space H. As described above, by placing the biomolecular material B together inside the silica shell layer 12, the biomolecular material B placed inside the accommodation space H can contact the nanoparticles 11, and as described below, surface-enhanced Raman scattering spectral signals can be obtained from the surface of the nanoparticles 11.

[0101] The "biomolecular material" in this specification refers to molecules and / or ions present in organisms, and can also refer to molecules and / or ions synthesized and / or produced during general biological processes such as cell division, morphogenesis, or development.

[0102] Biomolecular material B includes not only proteins, carbohydrates, lipids, nucleic acids, and inorganic substances, but also low-molecular-weight substances such as primary metabolites, secondary metabolites, and natural products, including both endogenous and exogenous substances. Biomolecular material B includes, for example, CA 19-9, amyloid beta40, and amyloid beta42 in plasma, but is not limited to the above examples.

[0103] On the other hand, as described in the structure, the structure 10 according to an embodiment may have nanoparticles 11 that non-specifically cause a surface plasmon resonance phenomenon with the biomolecular material B on the surface to form a peak. At this time, as an alternative embodiment, refer to Figure 1e , different biomolecular materials B may be present inside the structure 10. With this structure, it is also possible to obtain a plurality of surface-enhanced Raman scattering spectral signals from one structure 10.

[0104] On the other hand, through multiple measurements, a database (DB) can be established for the plurality of surface-enhanced Raman scattering spectral signals obtained, and specific peaks of diseases can be measured based on the signals in the established database. Further, if applied to diagnostic kits, etc., it can achieve the effect of accurately diagnosing diseases in a short time.

[0105] In one embodiment, the structure 10 may include two or more types of biomolecular materials B. By separating and extracting the upper or lower region of the biological solution treated with the pretreatment solution, the dimension of the total surface-enhanced Raman scattering spectral signals obtained can be diversified, enabling the establishment of a more detailed database for multiple different biomolecular materials B. Ultimately, a structure 10 that can obtain surface-enhanced Raman scattering spectral signals more accurately can be provided.

[0106] As described above, referring again to Figures 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 inside, and the nanoparticles 11 and the biomolecular material B may be placed in a region inside the accommodation space H of the silica shell layer 12.

[0107] When nano-sized particles are dispersed in a liquid or the like, they may aggregate due to the interaction (e.g., attractive force) between the particles in the dispersion liquid 20. However, in the case of having the silica shell layer 12 surrounding each nanoparticle 11 as described in the structure, the silica shell layer 12 can prevent the interaction between the nanoparticles 11, thereby preventing the occurrence of inter-particle aggregation. Furthermore, it can also provide the effect of stable measurement results by enhancing the surface-enhanced Raman scattering spectral signal or improving the reproducibility of the surface-enhanced Raman scattering spectral signal.

[0108] In addition, the structure 10 according to an embodiment does not necessarily require a substrate, position, or probe that specifically binds to the biomolecular material B. Therefore, when the biomolecular material B is located on the surface of the nanoparticle 11 or in contact with the surface of the nanoparticle 11, the surface-enhanced Raman scattering spectral signal of a large amount of biomolecular material B distributed inside the structure 10 can be measured.

[0109] As described above, the measured surface-enhanced Raman scattering spectral signal can be used to establish a database, and ultimately provide a diagnostic kit that can diagnose diseases even without biomarkers.

[0110] In one embodiment, the silica shell layer 12 may refer to a shell layer mainly formed of silicon (Si) and oxygen (O). The material thereof is not particularly limited, and as long as it is a silica shell layer 12 that can be easily selected by those skilled in the art, it should be understood to fall within the scope of the claims of the present invention. For example, there is a silica shell layer 12 formed by the polymerization reaction of TEOS, but it is not limited to the above example.

[0111] In one embodiment, the thickness of the silica shell layer 12 can be 20 to 30 nm, and as another alternative embodiment, it can be 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, 23.5 to 28, 23.5 to 27, 23.5 to 26, 23.5 to 25, or 23.5 to 24.

[0112] Additionally, in an alternative embodiment, the thickness of the silica shell layer 12 can be 23.28 nm or 21.85 nm, and the thickness as described above can vary depending on whether ethanol pretreatment is performed.

[0113] The composition for surface - enhanced Raman scattering spectroscopy is described below.

[0114] Figure 2a Schematic diagram of the diagnostic solution used in the diagnostic method provided according to one embodiment, Figure 2b For Figure 2a An enlarged view of region A in

[0115] See Figure 2a According to another embodiment of the present invention, a composition for surface - enhanced Raman scattering spectroscopy (hereinafter referred to as "diagnostic solution") is provided, which includes a dispersion liquid 20 and a structure body 10 for surface - enhanced Raman scattering spectroscopy (hereinafter referred to as "structure body") dispersed in the dispersion liquid 20. The composition can be used as the diagnostic solution 1 for surface - enhanced Raman scattering spectroscopy.

[0116] In one embodiment, the structure body 10 can be dispersed in the dispersion liquid 20 of the diagnostic solution 1, and a more specific and detailed description of the structure body 10 is replaced by the description in the foregoing embodiment.

[0117] In one embodiment, the dispersion liquid 20 may refer to a liquid-type substance in which nano-scale particles can be dispersed. The dispersion liquid 20 is not particularly limited, and any liquid-type substance capable of dispersing the nano-particles 11 that can be easily selected by those skilled in the art of this technology should be understood to fall within the scope of the claims of the present invention. On the other hand, the dispersion liquid 20 may include, for example, water (H2O), ethanol (EtOH), etc., but is not limited to the above examples.

[0118] In one embodiment, the diagnostic solution 1 may further include an additive. The additive includes, for example, a surfactant. The surfactant can make the nano-scale particles and / or the aforementioned structure 10 more uniformly dispersed in the dispersion liquid 20.

[0119] In one embodiment, the surfactant is not particularly limited, and a cationic surfactant and / or an anionic surfactant can be used. In addition, it should be understood that any surfactant that can be easily selected by those skilled in the art of this technology belongs to the scope of the claims of the present invention. On the other hand, the surfactant includes CTAB, CTAC, etc., but is not limited to the above examples.

[0120] Next, a manufacturing method of a structure 10 for surface-enhanced Raman scattering spectroscopy and a diagnostic solution 1 for surface-enhanced Raman scattering spectroscopy will be introduced.

[0121] Figure 3a and Figure 3b is a flowchart of a manufacturing method of a structure for surface-enhanced Raman scattering spectroscopy according to an embodiment.

[0122] See Figure 3a , to solve the above technical problems, an embodiment of the present invention provides a manufacturing method of a structure for surface-enhanced Raman scattering spectroscopy (hereinafter simply referred to as "manufacturing method of the structure"), which includes: a step S10 of forming a solution in which nano-particles 11 are dispersed and a step S20 of forming a silica shell layer 12 surrounding the nano-particles 11.

[0123] In addition, in an optional embodiment, the manufacturing method of the structure can perform the process of growing nano-particles 11 in a solution and forming a silica shell layer 12 surrounding the nano-particles 11, so as to provide a manufacturing method of a diagnostic solution 1 for surface-enhanced Raman scattering spectroscopy together.

[0124] In one embodiment, step S10 of forming a solution in which nanoparticles 11 are dispersed may include: a step of mixing a first solution containing a first precursor compound and a second solution containing a second precursor compound to form a mixed solution, wherein the first precursor compound includes a first plasmonic metal and the second precursor compound includes a second plasmonic metal.

[0125] 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 along the outer surface of the core 111 into a second plasmonic metal shell layer 112. By the manner described above, core-shell nanoparticles 11 can be formed.

[0126] A more specific and detailed description of the first plasmonic metal and the second plasmonic metal is replaced by the description in the foregoing embodiment.

[0127] On the other hand, the step of forming the core-shell nanoparticles 11 is not particularly limited, and any method that can be easily selected by those skilled in the art with ordinary knowledge in this technical field should be understood to fall within the scope of the claims of the present invention.

[0128] In one embodiment, the first precursor compound contains a first plasmonic metal and may refer to a compound capable of forming nanoscale nanoparticles containing the first plasmonic metal. This is not particularly limited, and it should be understood that any precursor compound of nanoparticles that can be easily selected by those skilled in the art belongs to the scope of the claims of the present invention. The first precursor compound includes, for example, HAuCl4·3H2O, but is not limited to the above example.

[0129] In one embodiment, the second precursor compound contains a second plasmonic metal and may refer to a compound capable of forming nanoscale nanoparticles containing the second plasmonic metal or the shell of nanoparticles. This is not particularly limited, and it should be understood that any precursor compound of nanoparticles that can be easily selected by those skilled in the art belongs to the scope of the claims of the present invention. The second precursor compound includes, for example, AgNO3, but is not limited to the above example.

[0130] In one embodiment, when mixing the first solution and the second solution, any one or more of a reducing agent and a surfactant may be mixed together. When forming the mixed solution, the first precursor compound forms the core 111 of the nanoparticles 11 through nano growth, and when mixed with one or more reducing agents and surfactants, the growth of the nanoparticles 11 can be further activated.

[0131] There are no particular limitations on the reducing agent and the surfactant, and it should be understood that as long as the reducing agent and the surfactant can be easily selected by those skilled in the art, they fall within the scope of the claims of the present invention. Examples thereof include NaBH4, CTAB, and CTAC, respectively.

[0132] In one embodiment, the method for manufacturing the structure may further include a step of mixing the mixed solution with a coating solution containing a water-soluble polymer compound. When the water-soluble polymer compound is mixed into the mixed solution containing the nanoparticles 11, the water-soluble polymer compound forms a polymer coating 113 in at least one region on the surface of the nanoparticles 11, which can prevent the phenomenon that the nanoparticles 11 aggregate together.

[0133] In one embodiment, examples of the water-soluble polymer compound include polyvinylpyrrolidone, polyvinyl alcohol, polysulfonate, hydroxyethyl cellulose, hydroxypropyl cellulose, cellulose acetate, and polyamide, etc., but are not limited to the above examples.

[0134] The polymer coating 113 can also enable biomolecular materials to be easily positioned on the metal surface, thereby providing better sensitivity when obtaining Raman signals. For example, blood contains various biomolecular materials such as nucleic acid, protein, organic matter, and inorganic matter. The polymer coating 113 can be coated on the surface of the nanoparticles as an intermediate substance for forming molecular interaction between the nanoparticles and the biomolecules, so that the above-mentioned biomolecular materials are located between the metal nanoparticles and the silica shell layer.

[0135] The biomolecular materials in the blood can be attached to the intermediate substance coated on the surface of the metal nanoparticles through intermolecular interaction, and the silica shell layer starts to form on the surface where the biomolecules are not attached. During the process of wrapping the metal nanoparticles, the biomolecules are fixed and do not fall off, and finally various biomolecular materials in the blood can exist in the accommodation space between the metal nanoparticles and the silica shell layer.

[0136] On the other hand, in order to form the polymer coating 113 on the surface of the nanoparticles 11, as an alternative embodiment, a mixed solution can be formed, and the coating solution can be mixed after a predetermined time.

[0137] In one embodiment, the method for manufacturing the structure may further include a step of mixing a coating solution containing a polymer compound containing the unit represented by the following Chemical Formula 1 into the mixed solution, and as an alternative embodiment, may further include a step of mixing a coating solution containing a water-soluble polymer compound containing the unit represented by the following Chemical Formula 1 into the mixed solution:

[0138] [Chemical Formula 1]

[0139]

[0140] In one embodiment, a method for manufacturing a structure may include the following steps: mixing a solution or a mixed solution in which nanoparticles 11 are dispersed with a third solution containing a biomolecular material B and a fourth solution containing a silica precursor compound. By mixing the biomolecular material B and the silica precursor compound in the solution containing the nanoparticles 11 so that they react, the nanoparticles 11 and the biomolecular material B can be distributed together inside the silica shell 12.

[0141] In one embodiment, a stirrer may be used to mix the biomolecular material B and the silica precursor compound in the solution containing the nanoparticles 11 and cause them to react, and as an alternative embodiment, a gyratory shaker or a seesaw shaker may be used for mixing to cause them to react.

[0142] Other more specific and detailed descriptions of the biomolecular material B can be replaced by the descriptions of the foregoing embodiments.

[0143] In one embodiment, the silica precursor compound may refer to a compound that can form the silica shell 12 after reaction, and there is no particular limitation as long as it is a silica precursor compound that can be easily selected by those skilled in the art of the present technology and should be understood to fall within the scope of the claims of the present invention. In this regard, the silica precursor compound includes, for example, TEOS, but is not limited to the examples.

[0144] See Figure 3b , in one embodiment, the third solution containing the biomolecular material B may be a solution treated with a pretreatment solution S15. The pretreatment solution may have a low-polarity substance, and as an alternative embodiment, it may include a substance with a polarity lower than that of water.

[0145] The third solution can be prepared by treating and / or processing a solution directly extracted from a living body (hereinafter referred to as a "biological solution"). Biological solutions include, for example, plasma, tissue fluid, and urine.

[0146] These biological solutions may use water (H2O) as a solvent. When the pretreatment solution with a low-polarity substance is mixed with the biological solution, biomolecular materials B with different molecular weights in the biological solution can be separated according to their molecular weights.

[0147] In one embodiment, substances with a polarity lower than that of water may include, but are not limited to, for example, aprotic substances, methanol, ethanol, acetone, DMSO, etc. As long as they are substances with a polarity lower than that of water that can be easily selected by those skilled in the art, they should be understood to fall within the scope of the claims of the present invention.

[0148] In one embodiment, the third solution may include two or more types of biomolecular materials B.

[0149] By separating and extracting the upper or lower region of the biological solution treated with the pretreated solution, the dimension of the obtained total surface-enhanced Raman scattering spectral signal can be diversified, so that a more detailed database can be established for the biomolecular material B. Finally, a structure 10 capable of diagnosing diseases more accurately can be provided.

[0150] Of course, the above embodiments of the present invention can be combined and applied to each other.

[0151] On the other hand, the present invention relates to a diagnostic method and a diagnostic system using surface-enhanced Raman scattering spectroscopy.

[0152] Figure 4a and Figure 4b is a flowchart of a diagnostic method using surface-enhanced Raman scattering spectroscopy provided according to an embodiment.

[0153] See Figure 4a , to solve the above technical problems, an embodiment of the present invention can provide a diagnostic method using surface-enhanced Raman scattering spectroscopy (hereinafter simply referred to as "the present diagnostic method"), which includes the following steps: an irradiation step S100 of irradiating light onto a diagnostic solution 1 containing a biomolecular material B to obtain a surface-enhanced Raman scattering spectral signal; and a step S200 of analyzing the obtained surface-enhanced Raman scattering spectral signal to diagnose a target disease.

[0154] In a living body suffering from a specific disease, biomolecular materials that specifically appear according to the specific disease may appear. As described above, a diagnostic method for diagnosing a target disease can be provided by using the specific surface-enhanced Raman scattering spectral signal (hereinafter referred to as "spectral signal") obtained from the biomolecular material.

[0155] As described below, the diagnostic method according to an embodiment may include a step (not shown) of obtaining a first spectral signal from a biological solution extracted from a patient suffering from a disease or a solution containing the biological solution. In addition, by obtaining the first spectral signals of patients suffering from various diseases, in the data storage unit 140 described later, spectral information related to the specific spectral signals according to the disease type can be established in a database.

[0156] Target diseases include pancreatic cancer, Alzheimer's disease, rheumatism, COVID-19 infection, influenza, Alzheimer's dementia, etc., but are not limited to the above examples.

[0157] In one embodiment, the biological solution refers to the liquids present in the human body, such as blood, plasma, tissue fluid, lymph fluid, urine, etc. In addition, the present invention is not limited to the above examples, but should be understood that any liquid in the human body that can be easily extracted by a person with ordinary knowledge in the technical field, a medical professional, or a medical assistant belongs to the scope of the claims of the present invention.

[0158] The diagnostic method according to one embodiment may include an irradiation step S100 of irradiating light onto the diagnostic solution 1 containing the biomolecular material B to obtain a spectral signal. Such a diagnostic solution is as described above, in which a structure for surface-enhanced Raman scattering spectroscopy may be dispersed, and a composition for surface-enhanced Raman scattering spectroscopy in which the above-mentioned structure 10 for surface-enhanced Raman scattering spectroscopy (hereinafter referred to as "structure") is dispersed may be used. In addition, these diagnostic solutions may be transported in the form in which they are generated, or may also be prepared at the location of diagnosing the disease. A more specific and detailed description of the diagnostic solution is replaced by the description of the structure 10 and the composition in which the structure 10 is dispersed in the foregoing embodiments.

[0159] In one embodiment, a second spectral signal may be obtained by irradiating light onto the diagnostic solution 1 containing the biomolecular material B. At this time, as described later, by comparing and analyzing the second spectral signal with the spectral information stored in the data storage unit 140, it is possible to diagnose which disease the diagnostic solution is for. In addition, as an alternative embodiment, during the comparison and analysis process, it is possible to identify which biomolecular material B is dissolved or dispersed in the diagnostic solution.

[0160] As described above, the diagnostic method provided according to one embodiment of the present invention obtains a spectral signal by directly irradiating light onto the liquid diagnostic solution, and since no separate drying process is required, the time required for diagnosis can be greatly reduced.

[0161] In addition, since the diagnostic method provided according to one embodiment diagnoses diseases, etc. by directly irradiating light L onto the liquid diagnostic solution 1, compared with the existing diagnostic methods, the problem of contamination of the experimental environment may not occur significantly. Further, the structure 10 described later dispersed in the liquid can maintain a similar environment even in repeated experiments, thereby improving reproducibility.

[0162] In addition, since the diagnostic solution 1 used in the present diagnostic method provided according to an embodiment can be stored and transported in a liquid state, the convenience of storing and transporting the diagnostic solution 1 can also be ensured. In addition, since it is manufactured in a liquid state, the diagnostic solution 1 can also be mass-produced.

[0163] In one embodiment, the light L irradiated onto the diagnostic solution 1 can be light having a wavelength band and intensity for surface-enhanced Raman scattering or Raman scattering, such as a laser. However, it is not limited to the above examples.

[0164] See Figure 2a , in one embodiment, the diagnostic solution 1 may include a dispersion liquid 20 and a structure body 10 dispersed in the dispersion liquid 20. With the above structure, in the present invention, without drying the diagnostic solution 1, spectral signals can be obtained by irradiating the light L onto the structure body 10 dispersed in a liquid state, thereby improving the reproducibility and accuracy of the signals.

[0165] In addition, the diagnostic solution 1 can be manufactured by mixing with a biological solution extracted from a diseased patient, so that the biological molecular material B contained in the structure body 10 included in the diagnostic solution 1 is included therein.

[0166] See Figure 2b , the structure body 10 may include a biological molecular material B in a region inside it, and when light is irradiated between the biological molecular material B distributed inside the structure body 10 and a region of the structure body 10, a surface plasmon phenomenon occurs, and a specific second spectral signal S according to the type of the biological molecular material B can be generated in a region of the structure body 10. By comparing and analyzing these second spectral signals S with the aforementioned spectral information, the target disease can be diagnosed.

[0167] In addition, referring again to Figure 2b , a region inside the structure body 10 may include different biological molecular materials B1 to B4. As described above, different biological molecular materials B1 to B4 will generate different spectral signals S1 to S4. In addition, the spectral signals S1 to S4 generated at this time are spectral signals specifically generated in the biological molecular materials B1 to B4.

[0168] By comparing and analyzing the spectral signals with the aforementioned spectral information, the target disease can be diagnosed. In addition, as an alternative embodiment, these spectral signals S1 to S4 can be analyzed to identify the biological molecular materials B1 to B4.

[0169] Just as Figures 1a to 1eThe structure of the structure body 10 is such that it has a silica shell layer 12, and the structure body 10 can be well dispersed in the dispersion liquid 20 without aggregating with each other. In addition, by distributing the nanoparticles 11 with plasmonic metal and the biomolecular material B together inside the silica shell layer 12, the surface-enhanced Raman scattering spectral signal can be more effectively generated between the nanoparticles 11 and the biomolecular material B.

[0170] Refer again to Figure 1e and Figure 2b , in one embodiment, the biomolecular material B may include two or more types of biomolecular materials B1 to B4, and as described above, when different types of biomolecular materials B are included, the surface-enhanced Raman scattering spectral signals S1 to S4 specific to the relevant biomolecular materials B can be generated.

[0171] In one embodiment, in the figure, the second plasmonic metal shell layer 112 is shown to have a regular hexahedron shape. However, in addition to having a regular hexahedron structure, the second plasmonic metal shell layer 112 may also have a regular polyhedron structure, and the regular polyhedron may include at least one of, for example, a regular octahedron, a regular dodecahedron, and a regular icosahedron.

[0172] 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 may be narrower than the width of the plasmon resonance peak on the surface of the first plasmonic metal.

[0173] As described in the structure, by making the width of the plasmon resonance peak on the surface of the second plasmonic metal narrower, better sensitivity can be provided when obtaining the surface-enhanced Raman scattering spectral signal when the structure body 10 is dispersed in the dispersion liquid 20.

[0174] At this time, as an alternative embodiment, 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.

[0175] Refer to Figures 1a to 1e , in one embodiment, the nanoparticles 11 may include a core 111, a second plasmonic metal shell layer 112, and a polymer coating 113 surrounding the second plasmonic metal shell layer 112. The polymer coating 113 imparts a negative charge to the surface of the nanoparticles 11 and can prevent the nanoparticles 11 from aggregating with each other.

[0176] With the described structure, during the manufacturing process of the structure 10, it is possible to prevent the nanoparticles 11 from aggregating with each other before the formation of the silica shell layer 12. Further, it is possible to more easily form the silica shell layer 12. For a more detailed description of the structure 10, the description of the foregoing related embodiments is used instead.

[0177] In one embodiment, the structure 10 can be formed by mixing and reacting a solution in which nanoparticles 11 are dispersed, a solution containing the biomolecular material B, and a solution containing a silica precursor compound. For a more detailed description of the manufacturing method of the structure 10, the description of the foregoing related embodiments is used instead.

[0178] In one embodiment, the solution containing the biomolecular material B can be, for example, a biological solution extracted from an organism, or a solution formed by mixing the biomolecular material B with a solvent, but is not limited to these examples. On the other hand, the solution containing the biomolecular material B can include a pretreatment solution, and the solution used in such a pretreatment process can be a solution with a lower polarity than water.

[0179] By means of the pretreatment process as described above, the solution containing the biomolecular material B can have various chemical or physical environments, and the dimensions of the total surface-enhanced Raman scattering spectral signals obtained can also be diversified, so that a more detailed database can be established for the biomolecular material B. Ultimately, a structure 10 capable of more accurately obtaining surface-enhanced Raman scattering spectral signals can be provided.

[0180] In one embodiment, substances with a lower polarity than water can include, but are not limited to, for example, aprotic substances, methanol, ethanol, acetone, DMSO, etc. As long as they are substances with a lower polarity than water that can be easily selected by those skilled in the art, they should be understood to fall within the scope of the claims of the present invention.

[0181] In one embodiment, the manufacturing method of the structure 10 is not particularly limited, and as long as it is a manufacturing method of the structure 10 that can be easily selected by those skilled in the art of the present technology field, it should be understood to fall within the scope of the claims of the present invention. For example, it can be manufactured using the principle of nanogrowth of metal particles in a solution, or by stirring a solution containing core-shell structured nanoparticles 11 and a solution containing TEOS and a biomolecular material. Preferably, the structure 10 can follow the manufacturing method of the structure and diagnostic method for surface-enhanced Raman scattering spectroscopy as described above.

[0182] See again Figure 4a, this diagnostic method may include step S200, which diagnoses the target disease by analyzing the second spectral signal obtained by the diagnostic solution 1 in step S100. At this time, as an optional embodiment, it may include step S220 of diagnosing the target disease by comparing and analyzing the second spectral signal obtained from the diagnostic solution with the spectral information S210 (see Figure 4b ).

[0183] As described above, in the body of a patient suffering from a specific disease, biomolecular materials that specifically appear according to the specific disease may be distributed. According to one embodiment, a specific spectral signal may be obtained based on the biomolecular material B inside the structure 10, and the target disease may be diagnosed by analyzing this signal. As an optional embodiment at this time, the biomolecular material B may be identified and used to diagnose the target disease.

[0184] The diagnostic method according to one embodiment may include a step (not shown) of obtaining a first spectral signal from a biological solution extracted from a patient suffering from a specific disease or a solution containing the biological solution. In addition, as an optional embodiment, a database may be established for the first spectral signal, so that the spectral information is stored in the data storage unit 140 or the like.

[0185] At this time, there is no particular limitation on the method of obtaining the first spectral signal, as long as it is a surface-enhanced Raman scattering method in the form of nanoparticles, substrates, liquid or solid, etc. In addition, any method of obtaining a surface-enhanced Raman scattering spectral signal that can be easily selected by those skilled in the art should be understood to fall within the scope of the claims of the present invention.

[0186] In addition, as an optional embodiment, the method may include the following steps: obtaining the spectral signal of the biomolecular material B, and thus establishing a database (DB) for the spectral signal of the biomolecular material B to store the spectral information. At this time, the spectral information for which the database is established may be stored in the data storage unit 140 in the embodiments described later.

[0187] In another optional embodiment, the step of establishing a database for the biomolecular material B may include: obtaining a spectral signal by using the target bacteria and / or viruses that cause any disease, and establishing a database for the biomolecular material B that causes any disease.

[0188] According to the type of disease, biomolecular materials B that specifically exist may be present in the body. By analyzing the types of biomolecular materials B from various angles, the target disease can be diagnosed. As described above, such biomolecular materials B include not only proteins and sugars, but also bacteria and viruses, etc.

[0189] On the other hand, in one embodiment, referring again toFigure 2b The biomolecular material B may include two or more different types of biomolecular materials B. Depending on the type of disease, the biomolecules specifically produced in vivo may also include two or more types of biomolecular materials B. By identifying the biomolecular material B in this way, the accuracy and reproducibility of target disease diagnosis can be further improved.

[0190] In one embodiment, the method for comparing and analyzing the first spectral signal and the second spectral signal may adopt methods such as comparing the intensity, consistency, and specific wave number of the verification peaks.

[0191] For example, when comparing the first spectral signal and the second spectral signal, in the case of pancreatic cancer patients, when obtaining the surface-enhanced Raman scattering signal, spectral signals with peaks generated at 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 and 435 cm -1 can be obtained. While in the diagnostic solution, when spectral signals are generated at 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 described above, the biomolecular material contained in the diagnostic solution as described above can be diagnosed as the biomolecular material belonging to pancreatic cancer patients.

[0192] In one embodiment, the present diagnostic method may further include a step of comparing with a control experimental group. It can be verified whether the spectral signal obtained using Diagnostic Solution 1 is obtained through the normal operation of the diagnostic system 100, and a control experimental group as described above can be formed using the same solvent as Diagnostic Solution 1.

[0193] On the other hand, the step of comparing with the control experimental group as described above is not particularly limited, and any method that can be selected by those skilled in the art with ordinary knowledge in this technical field should be understood to fall within the scope of the claims of the present invention.

[0194] Figures 5a to 5c is a block diagram showing a diagnostic system 100 provided according to an embodiment.

[0195] To solve the above technical problems, another embodiment of the present invention provides a disease diagnostic system (hereinafter referred to as "the present diagnostic system") 100 using surface enhanced Raman scattering spectroscopy.

[0196] See Figure 5a Figure 5a , the diagnostic system 100 provided according to an embodiment may include: a container unit 110 for placing a diagnostic solution 1; an irradiation unit 120 for irradiating light to the diagnostic solution 1 placed in the container unit 110 to obtain a spectral signal from a region in the structure 10; and a diagnostic unit 130 for analyzing the obtained spectral signal.

[0197] In one embodiment, the present diagnostic system 100 may further include a manufacturing unit for manufacturing the structure 10. The manufacturing unit may manufacture the structure 10 containing the biomolecular material B in an internal region.

[0198] At this time, in an alternative embodiment, the manufacturing unit may manufacture the diagnostic solution 1 in which the structure 10 is dispersed. As described above, the manufactured diagnostic solution 1 may be used for the container unit 110, the irradiation unit 120, etc. described later.

[0199] On the other hand, the method for manufacturing the structure 10 in the manufacturing unit as described above is not particularly limited, and a more specific and detailed description of the manufacturing method of the structure 10 is replaced by the description in the foregoing embodiment.

[0200] In one embodiment, the diagnostic solution 1 may include the structure 10. The present diagnostic system 100 may directly irradiate light to the diagnostic solution 1 through the irradiation unit 120 to obtain a spectral signal from a region of the structure 10 distributed in the diagnostic solution 1, and may diagnose a target disease by analyzing the spectral signal in the diagnostic unit 130.

[0201] On the other hand, as described above, the structure 10 may include the biomolecular material B, and a more specific and detailed description thereof is replaced by the description in the foregoing embodiment.

[0202] In one embodiment, the container unit 110 may be configured to place the diagnostic solution 1 therein, and may irradiate light to the diagnostic solution 1 placed in the container unit 110 to generate a spectral signal in a region of the structure 10. Ensuring the sufficient placement of the diagnostic solution 1 is a preferred method. As an alternative embodiment, the container unit 110 for placing the diagnostic solution 1 may be at least 200 μL or more, but of course it is not limited to this size.

[0203] On the other hand, the container part 110 described above is not particularly limited and can be easily selected by those skilled in the art with ordinary knowledge in this technical field, and it should be understood that any container that can hold a liquid to generate a spectral signal falls within the scope of the claims of the present invention.

[0204] In one embodiment, the irradiation part 120 is configured to irradiate light onto the diagnostic solution 1. When light is irradiated onto the diagnostic solution 1, a spectral signal is generated in a region within the structure 10 distributed in the diagnostic solution 1.

[0205] At this time, as an alternative embodiment, the structure 10 may include nanoparticles 11, a biomolecular material B, and a silica shell surrounding them. In addition, a spectral signal may appear near the surface where the biomolecular material B and the nanoparticles 11 meet. On the other hand, more specific and detailed descriptions of the structure 10 can be replaced by the descriptions of the foregoing embodiments.

[0206] Refer again to Figure 5a In one embodiment, the irradiation part 120 may include an irradiation unit that emits light. The irradiation unit is not particularly limited, and as an alternative embodiment, it may include a laser irradiation unit.

[0207] Refer to Figure 5b In one embodiment, the diagnostic part 130 can diagnose a target disease by comparing and analyzing the spectral information and the second spectral signal in the above embodiments. As an alternative embodiment, the biomolecular material B contained in the diagnostic solution 1 can be identified during this process.

[0208] On the other hand, when using the information of the spectral signal in the diagnostic part 130, the spectral information of the first spectral signal stored in the data storage part 140 described later can be used.

[0209] In one embodiment, the diagnostic part 130 can use an algorithm to compare and / or analyze the peak wave number consistency, peak intensity, consistency, etc. between the spectral information and the second spectral signal. Through this process, the first spectral signal can be searched, and the target disease specifically corresponding to the searched first spectral signal can be diagnosed.

[0210] Refer to FIG. 3c. In one embodiment, the diagnostic part 130 may include: a first diagnostic part 130a for searching for the first spectral signal corresponding to the second spectral signal in the spectral information; and a second diagnostic part 130b for diagnosing a disease using the searched first spectral signal.

[0211] Refer again to Figure 5b, in one embodiment, the diagnostic system 100 may include a data storage unit 140 that acquires spectral signals to store spectral information.

[0212] In one embodiment, the data storage unit 140 may store the spectral information of the above-mentioned first spectral signal. The diagnostic unit 130 may search for the first spectral signal corresponding to the second spectral signal in the spectral information by comparing the spectral information stored in the data storage unit 140 with the second spectral signal. In addition, a specific disease can be searched out through the first spectral signal. Thus, the diagnostic unit 130 can diagnose the target disease.

[0213] Referring again Figure 5b and Figure 5c , in one embodiment, the data storage unit 140 may include a first data storage unit 140a that stores the spectral information of the first spectral signal. The spectral information includes, for example, a spectrogram of a surface-enhanced Raman scattering spectral signal of a specific disease, spectral intensity, a set value of the irradiation unit, etc., but is not limited to the above examples.

[0214] In one embodiment, the data storage unit 140 may include a second data storage unit 140b that stores information about diseases and the first spectral signal. The diagnostic unit 130 may use the information recorded in the second data storage unit 140b to diagnose the target disease.

[0215] As an alternative embodiment, the data storage unit 140 may establish a database for the information of the biomolecular material B. The information of the spectral signal of the biomolecular material B as described above may be a specific Raman scattering spectral signal generated according to, for example, the types of proteins, bacteria, viruses, or inorganic substances. The description of how to obtain such information can be replaced by the description in the foregoing embodiments.

[0216] On the other hand, in one embodiment, communication may be performed between the data storage unit 140 and the diagnostic unit 130 using a wired / wireless communication method, and a diagnostic system 100 for diagnosing diseases by a near / long-distance method can be provided through the method described above. On the other hand, there are no particular limitations on the wired / wireless communication method as described above, and any communication method that can be easily selected by those skilled in the art with ordinary knowledge in this technical field should be understood to fall within the scope of the claims of the present invention.

[0217] Of course, the above embodiments of the present invention can be combined and applied to each other.

[0218] Hereinafter, manufacturing examples, embodiments, and experimental examples of the present invention will be described. However, it should be understood that these manufacturing examples, embodiments, and experimental examples are for more specifically illustrating the composition and effects of the present invention, and the scope of the present invention is not limited thereto.

[0219] Example

[0220] Example 1 - Structure Containing Nanoparticles with Core - Shell Structure

[0221] In this Example 1, a structure for surface - enhanced Raman scattering spectroscopy and a composition for surface - enhanced Raman scattering spectroscopy in which the structure is dispersed were fabricated. The specific fabrication process is in the order described below.

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

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

[0224] To form CTAB - coated gold nanoparticles with a size of 2 nm to 3 nm, 9.75 ml of a 100 mM CTAB solution was prepared, 250 μl of a 10 mM HAuCl4·3H2O solution and 600 μl of a 10 mM NaBH4 solution were added, and after stirring for 2 minutes, the reaction was carried out for 3 hours.

[0225] Subsequently, to grow the formed CTAB - coated 2 nm to 3 nm gold nanoparticles into 30 nm gold nanoparticles, 16 mL of a 200 mM CTAC solution was prepared, and then 16 mL of a 0.5 mM HAuCl4·3H2O solution and 16 mL of a 0.8 mM ascorbic acid solution were added to 40 μL of the previously prepared CTAB - capped AuNP seed, and the reaction was carried out for 7 days.

[0226] Subsequently, to form a second plasmonic metal shell 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 carried out for 40 minutes. After that, after centrifugation, the solution was redispersed in triple - distilled water to form gold - silver core - shell nanoparticles.

[0227] (2) Coating the surface of core-shell structured nanoparticles with PVP

[0228] After preparing 10 mL of a previously formed gold-silver core-shell nanoparticle solution, 0.5 mL of a 1 v / v% SDS solution and 5 mL of a 2.5% (w / v) PVP solution were added, and the reaction was carried out for 10 minutes. Then, after centrifugation, it was redispersed in distilled water to form core-shell nanoparticles with a PVP coating layer on the surface.

[0229] (3) Formation of biomolecular materials and silica shell layers

[0230] In order to form core-shell nanoparticles with a previously formed PVP coating layer, biomolecular materials near the nanoparticles, and a silica shell layer surrounding them, the following procedures were carried out in sequence. At this time, in Example 1.1, a structure for surface-enhanced Raman scattering spectroscopy was fabricated by pre-treating plasma with ethanol, while in Example 1.2 below, a structure for surface-enhanced Raman scattering spectroscopy was fabricated using plasma without a pre-treatment process.

[0231] Example 1.1 - Pretreatment

[0232] After preparing 10 μL of plasma, 20 μL of deionized water and 70 μL of ethanol were added, and the reaction was carried out for 10 minutes to pre-treat the plasma.

[0233] 17.5 μL of core-shell nanoparticles with a PVP coating layer, 20 μL of the plasma supernatant pre-treated with ethanol, 44.5 μL of deionized water, and 20 μL of a 0.05 v / v% TEOS solution were added, and the reaction was carried out for 30 minutes. Then, 20 μL of a 10 v / v% DMA solution was added to produce a composition for surface-enhanced Raman scattering spectroscopy in which a structure for surface-enhanced Raman scattering spectroscopy was dispersed.

[0234] Example 1.2 - Without Pretreatment

[0235] 17.5 μL of core-shell nanoparticles with a PVP coating layer, 2 μL of plasma not pre-treated 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 carried out for 30 minutes. Then, 20 μL of a 10 v / v% DMA solution was added to produce a composition for surface-enhanced Raman scattering spectroscopy in which a structure for surface-enhanced Raman scattering spectroscopy was dispersed.

[0236] Experimental Example 1 - Verification of the Structure

[0237] Experimental Example 1 is an experiment to verify the state of the structure for surface-enhanced Raman scattering spectroscopy fabricated through the aforementioned Example 1.1 and Example 1.2.

[0238] Experimental Example 1.1 - Verification of the Size of Nanoparticles

[0239] In this Experimental Example 1.1, an experiment was conducted to verify the size of nanoparticles. The specific experimental method was to measure the size of nanoparticles using dynamic light scattering and transmission electron microscopy (TEM) imaging.

[0240] Measuring the size of nanoparticles using dynamic light scattering showed that the diameter of spherical gold nanocores was 31 nm and the PI was 0.16. The diameter of hexahedral gold-silver core-shell nanoparticles was 47.5 nm and the PI was 0.25.

[0241] Refer to Figure 6a the TEM image of

[0242] Experimental Example 1.2 - Verification of the Silica Shell

[0243] In this Experimental Example 1.2, an experiment was conducted to verify whether a silica shell was formed on the surface of the structure. The specific experimental method was to measure the size of nanoparticles using dynamic light scattering and TEM imaging.

[0244] The diameter of nanoparticles with a silica shell formed after ethanol pretreatment measured using dynamic light scattering was 90.25 nm and the PI was 0.35, while the diameter of nanoparticles with a silica shell formed without ethanol pretreatment was 87.4 nm and the PI was 0.31. The thickness of the silica shell after ethanol pretreatment was 23.28 nm, and the thickness of the silica shell without pre-ethanol treatment was 21.85 nm.

[0245] Refer to Figure 6b the TEM image of

[0246] Experimental Example 1.3 - Verification of Biomolecular Materials

[0247] In this Experimental Example 1.3, the Raman scattering spectra were compared when biomaterials were present and absent inside the structure.

[0248] Specific experimental methods confirmed the differences in Raman spectra between the silica shells (red) formed by adding plasma samples to PVP-coated silver (Ag) nanocube particles and the silica shells (blue) formed without adding plasma samples.

[0249] Figure 7 To show the experimental result graph of Experimental Example 1.3.

[0250] See Figure 7 , when verifying the differences in Raman spectra in the nanostructure with and without plasma samples, i.e., in the presence and absence of biomaterials, it can be confirmed that many new peaks appear or some peaks disappear in the spectra. For example, when plasma is present, peaks at 1170 cm -1 and 1520 cm -1 are newly generated, while peaks at 930 cm -1 and 1130 cm -1 disappear.

[0251] Experimental Example 2 - Verification experiment of Raman spectra with or without pretreatment

[0252] In this Experimental Example 2, through the above-mentioned Example 1.1 and Example 1.2, experiments were conducted to verify the differences in Raman spectra according to whether ethanol pretreatment was carried out.

[0253] The specific experimental method was to stack the silica shell with plasma and nanoparticles together, mix 70% ethanol and plasma at a ratio of 9:1 (v / v), let it stand for 10 minutes, and then stack the silica shell with nanoparticles together to obtain their respective Raman spectra for comparison.

[0254] Figure 8 To show the Raman spectra of the experimental results of this Experimental Example 2.

[0255] See Figure 8 , comparing the case without pretreatment (blue) and the case with ethanol pretreatment (orange), it can be seen that in the case of pretreatment, peaks are located at 650 cm -1 and 880 cm -1 , while in the case without pretreatment, peaks are located at 1280 cm -1 and 1390 cm -1 .

[0256] Experimental Example 3 - Verification of surface-enhanced Raman scattering spectral signals for diagnosing target diseases

[0257] In this Experimental Example 3, using the diagnostic solution manufactured in the above-mentioned Example 1, surface-enhanced Raman scattering spectral signals were obtained.

[0258] Place the diagnostic solution in a 96-well plate, irradiate it with a 532 nm laser, and use a Raman spectrometer to obtain the scattered light. The results of this verification are as Figure 9 shown.

[0259] Experimental Example 4 - Verification of the Diagnostic Error Rate of the Target Disease

[0260] In this Experimental Example 4, the spectral signals obtained through the above Experimental Example 3 are used to diagnose the target disease. By verifying the peaks appearing at 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 and 435 cm -1 , normal people and cancer patients are distinguished with a precision of 0.931, a sensitivity of 0.800, and a specificity of 0.800.

[0261] The above description of the present invention is only for descriptive purposes, and those skilled in the art with ordinary knowledge in the field can understand that, without changing the technical idea or essential features of the present invention, the present invention can be easily modified into other specific forms. Therefore, it should be understood that the above embodiments are exemplary in all aspects and not restrictive. For example, each component described as a single entity can be implemented in a distributed manner, and similarly, the components described as distributed can be implemented in a combined manner.

[0262] The scope of the present invention is determined by the following claims, and all changes or modifications derived from the meaning and scope of the claims and equivalent concepts should be construed as being included within the scope of the present invention.

Claims

1. A structure for surface-enhanced Raman scattering spectroscopy, comprising: A silica shell having an accommodation space inside; And nanoparticles having a plasmonic metal, the nanoparticles being distributed in a region of the accommodation space inside the silica shell.

2. The structure for surface-enhanced Raman scattering spectroscopy according to claim 1, wherein The nanoparticles include: a core containing a first plasmonic metal; And a second plasmonic metal shell surrounding the core and containing a second plasmonic metal, The second plasmonic metal shell has a symmetric structure with respect to the center of the second plasmonic metal shell.

3. The structure for surface-enhanced Raman scattering spectroscopy according to claim 1, wherein The structure includes a biomolecular material placed in the accommodation space.

4. The structure for surface-enhanced Raman scattering spectroscopy according to claim 2, wherein The first plasmonic metal and the second plasmonic metal each independently include any one or more selected from the group consisting of Au, Ag, Cu, Al, W, Pt, Ni, and Pd.

5. The structure for surface-enhanced Raman scattering spectroscopy according to claim 2, 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. The structure for surface-enhanced Raman scattering spectroscopy according to claim 2, wherein The second plasmonic metal shell has a regular polyhedron structure.

7. A method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy, comprising the following steps: Preparing a solution in which nanoparticles are dispersed; And forming a silica shell surrounding the nanoparticles dispersed in the solution.

8. The method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy according to claim 7, wherein The step of preparing a solution in which nanoparticles are dispersed includes: mixing a first solution containing a first precursor compound with a second solution containing a second precursor compound to form a mixed solution, wherein the first precursor compound contains a first plasmonic metal and the second precursor compound contains a second plasmonic metal, Wherein the nanoparticles include: a core and a second plasmonic metal shell, the core contains a first plasmonic metal, the second plasmonic metal shell surrounds the core and has a second plasmonic metal, The second plasmonic metal shell has a symmetric structure with respect to the center of the second plasmonic metal shell.

9. The method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy according to claim 8, It further includes the step of mixing a coating solution containing a water-soluble polymer compound into the mixed solution.

10. The method for manufacturing a structure for surface-enhanced Raman scattering spectroscopy according to claim 8, wherein The step of forming the silica shell layer includes the following steps: mixing the mixed solution, a third solution containing a biomolecular material, and a fourth solution containing a silica precursor compound. The silica shell layer enables at least a part of the nanoparticles and the biomolecular material to be distributed together inside it.

11. A disease diagnosis method using surface-enhanced Raman scattering spectroscopy, which includes the following steps: Obtaining a first surface-enhanced Raman scattering spectral signal from a biological solution extracted from a patient; Irradiating light onto a diagnostic solution in which the structure for surface-enhanced Raman scattering spectroscopy according to any one of claims 1 to 6 is dispersed, to obtain a second surface-enhanced Raman scattering spectral signal in a region of the structure for surface-enhanced Raman scattering spectroscopy; And comparing the first surface-enhanced Raman scattering spectral signal with the second surface-enhanced Raman scattering spectral signal to identify the disease of the patient.

12. The disease diagnosis method using surface-enhanced Raman scattering spectroscopy according to claim 11, wherein The structure for surface-enhanced Raman scattering spectroscopy includes two or more types of biomolecular materials.

13. The disease diagnosis method using surface-enhanced Raman scattering spectroscopy according to claim 11, wherein The diagnosis method further includes the following steps: Establishing a database of the first surface-enhanced Raman scattering spectral signal obtained from the biological solution extracted from the patient.

14. The disease diagnosis method using surface-enhanced Raman scattering spectroscopy according to claim 11, wherein The diagnosis method further includes the following steps: Obtaining the surface-enhanced Raman scattering spectral signal of the biomolecular material in the structure for surface-enhanced Raman scattering spectroscopy to establish a database of the information of the surface-enhanced Raman scattering spectral signal of the biomolecular material.

15. A disease diagnosis system using surface-enhanced Raman scattering spectroscopy, comprising: A container part for placing the diagnostic solution; An irradiation part that irradiates light onto the diagnostic solution placed in the container part and generates a surface-enhanced Raman scattering spectral signal in a region of the structure for surface-enhanced Raman scattering spectroscopy according to any one of claims 1 to 6 contained in the diagnostic solution; And A diagnosis part that analyzes the generated surface-enhanced Raman scattering spectral signal.

16. The disease diagnosis system using surface-enhanced Raman scattering spectroscopy according to claim 15, wherein The structure for surface-enhanced Raman scattering spectroscopy includes two or more types of biomolecular materials.

17. The disease diagnosis system using surface-enhanced Raman scattering spectroscopy according to claim 15, wherein The diagnosis system further includes: a data storage part for obtaining the surface-enhanced Raman scattering spectral signal to store the spectral information of the surface-enhanced Raman scattering spectral signal.

Images