KR-20260062736-A - Reactive oxygen species measuring structure, its manufacturing method, reactive oxygen species measuring device, and its manufacturing method

KR20260062736AKR 20260062736 AKR20260062736 AKR 20260062736AKR-20260062736-A

Abstract

A method for manufacturing an active oxygen measuring structure is provided, comprising the steps of: manufacturing a target recognition structure that selectively recognizes a target by combining a single-walled carbon nanotube (SWCNT) and a single-stranded DNA (ssDNA); and combining the target recognition structure with a cell-friendly polymer, wherein the cell-friendly polymer provides a cell-friendly interface to the target recognition structure.

Inventors

조수연
조영욱
김진웅
백휘라

Assignees

성균관대학교산학협력단

Dates

Publication Date: 20260507
Application Date: 20241029

Claims (14)

A step of manufacturing a target recognition structure that selectively recognizes a target by combining a single-walled carbon nanotube (SWCNT) and a single-stranded DNA (ssDNA); and The method includes the step of combining the above-mentioned target recognition structure with a cell-philic polymer, wherein The above-mentioned cytophilic polymer is, A method for manufacturing an active oxygen measuring structure, comprising providing a cell-friendly interface to the above-mentioned target recognition structure.
In Article 1, The step of manufacturing the above target recognition structure is, A step of mixing the single-walled carbon nanotube and the single-stranded DNA to combine the single-walled carbon nanotube and the single-stranded DNA; and The method includes a step of forming a corona phase by π-π stacking interactions formed by the bonding between the single-walled carbon nanotube and the single-stranded DNA, wherein The above target recognition structure is, A method for manufacturing an active oxygen measuring structure comprising selectively recognizing a target through the above corona phase.
In Article 1, The step of providing the cell-friendly interface described above is: The method comprises the step of mixing the above-mentioned target recognition structure and the above-mentioned cytophilic polymer, wherein The above-mentioned cytophilic polymer includes a positively charged functional group, and A method for manufacturing an active oxygen measuring structure, wherein the cell-friendly interface comprises being more neutral than the target recognition structure.
In Article 1, The above target recognition structure and the above cell-philic polymer are, A method for manufacturing an active oxygen measuring structure comprising mixing in a mass ratio greater than 10:0 and less than 10:10.
In Article 1, The above target includes hydrogen peroxide ( H₂O₂ ), and A method for manufacturing an active oxygen measuring structure, wherein the above single-stranded DNA comprises a (CCCT) 7 sequence.
A target recognition structure comprising a single-walled carbon nanotube and a single-stranded DNA coupled to the single-walled carbon nanotube, wherein the single-walled carbon nanotube selectively recognizes a target; and A cell-philic polymer combined with the above-mentioned target recognition structure, comprising The above-mentioned cytophilic polymer is, A reactive oxygen species measuring structure comprising providing a cell-friendly interface to the above-mentioned target recognition structure.
In Article 6, The above target recognition structure is, Selectively recognizing the target through a corona phase formed by π-π stacking interactions formed by the binding between the single-walled carbon nanotube and the single-stranded DNA, wherein The above target is an active oxygen measuring structure comprising hydrogen peroxide.
In Article 6, The above target recognition structure includes a negative charge, and The above-mentioned cytophilic polymer is an active oxygen measuring structure comprising a positively charged functional group.
In Article 8, The above-mentioned cytophilic polymer is, An active oxygen measuring structure comprising providing affinity between the target recognition structure and the cell through the positively charged functional group.
In Article 6, An active oxygen measuring structure comprising, when reacting with the above-mentioned target, a decrease in near-infrared fluorescence due to electron transfer being used as a target detection signal.
In Article 6, The above-mentioned cytophilic polymer is, An active oxygen measuring structure comprising at least one selected from the group including poly-l-lysine (PLL) and poly-d-lysine (PDL).
Step of preparing the substrate; A step of forming a functional layer by coating a functional material on the substrate to ensure uniformity and stability of the material formed on the substrate; and The method comprises the step of forming a sensing layer by coating a target recognition structure comprising a single-strand DNA coupled to a single-walled carbon nanotube, and an active oxygen measurement structure comprising a cytophilic polymer coupled to the target recognition structure, on the above functional layer. A method for manufacturing an active oxygen measuring device, comprising forming a corona phase that selectively recognizes a target by means of π-π stacking interactions formed by the bonding between the single-walled carbon nanotube and the single-stranded DNA.
Substrate; A sensing layer formed on the substrate, comprising a target recognition structure including a single-strand DNA coupled to a single-walled carbon nanotube, and an active oxygen measuring structure including a cytophilic polymer coupled to the target recognition structure; and A functional layer formed between the substrate and the sensing layer, providing uniformity and stability to the sensing layer on the substrate, comprising: The above-mentioned cell-friendly polymer provides a cell-friendly interface to the above-mentioned target recognition structure, and An active oxygen measuring device comprising the above target recognition structure selectively recognizing a target through the cell-friendly interface.
In Article 13, The above target includes hydrogen peroxide, and An active oxygen measuring device comprising a reduction in near-infrared fluorescence due to electron transfer when reacting with the above-mentioned target, wherein such reduction is used as a target detection signal.

Description

Reactive oxygen species measuring structure, its manufacturing method, reactive oxygen species measuring device, and its manufacturing method The present application relates to a reactive oxygen species measuring structure, a method for manufacturing the same, a reactive oxygen species measuring device, and a method for manufacturing the same. More specifically, it relates to a reactive oxygen species measuring structure, a method for manufacturing the same, a reactive oxygen species measuring device, and a method for manufacturing the same, which has excellent affinity for cells and can stably and selectively detect a target in said cells while minimizing damage to said cells in a non-labeled manner. Photoaging refers to the phenomenon in which the skin ages due to ultraviolet rays irradiated onto the skin. Due to photoaging, the skin becomes rough, loses elasticity, becomes dry, and thickens; pigmentation such as freckles, melasma, and blemishes occurs; and the skin's capillaries expand, causing redness. Accordingly, various methods for measuring photoaging have been developed in the past. For example, Korean Registered Patent Publication No. 10-1756478 discloses a kit for determining a subject's skin condition and genetic predisposition to ultraviolet radiation (UVR) damage, comprising a material for collecting a skin sample, a primer or probe for performing detection of mitochondrial DNA (mtDNA) aberration and one or more melanocortin 1 receptor (MC1R) variants, and a reagent for performing detection of mtDNA aberration and one or more MC1R variants, wherein the mtDNA aberration is a 3895 bp mtDNA deletion between nucleotides 547 and 4443 of the mtDNA genome of SEQ ID NO. 1, and the one or more MC1R variants are selected from the group consisting of D84E, R142H, R151C, R160H, D294H, V60L, and V92M. As another example, Korean Registered Patent Publication No. 10-2573314 discloses a method for manufacturing carbon quantum dots that exhibit color change characteristics based on the cumulative amount of exposure to ultraviolet light, comprising solvothermal synthesis of a blue inkjet printer dye, urea, and an organic solvent in a high-pressure reactor at a temperature of 180°C or higher and 250°C or lower for 3 hours or more, wherein the organic solvent comprises one or more selected from DMF (Dimethylformamide), ethanol, cyclohexane, toluene, THF (Tetrahydrofuran), and benzene. However, conventional photoaging monitoring technology may rely primarily on fluorescence analysis using organic dyes and methods tracking changes in reactive oxygen species using microscopes. Conventional methods using organic dyes have been used to accurately track reactive oxygen species release in spacetime in 2D and/or 3D skin models, but they may have limitations in analyzing the dynamic characteristics of photoaging induced by routine low-intensity ultraviolet radiation. In addition, conventional photoaging monitoring technology may rely on low-resolution static analysis at the minute level, and may have the disadvantage that real-time precise measurement of reactive oxygen species at the millisecond level is difficult due to photobleaching of fluorescent dyes, and immediate monitoring of intercellular interactions is difficult. In addition, conventional methods may have difficulty determining accurate reactive oxygen species concentrations and spatial dynamics due to signal variability resulting from the intracellular absorption process of dyes, and since stress can be applied to cells by dye labeling alone, it may be difficult to observe pure stress caused by ultraviolet rays. Accordingly, a new method for measuring photoaging is required. FIG. 1 is a drawing for explaining a method for manufacturing an active oxygen measuring structure according to an embodiment of the present application. FIG. 2 is a drawing for explaining the step of manufacturing a target recognition structure according to an embodiment of the present application. FIG. 3 is a drawing for explaining a method for selectively recognizing a target according to an embodiment of the present application. FIG. 4 is a diagram illustrating the step of combining a target recognition structure and a cell-philic polymer according to an embodiment of the present application. FIG. 5 is a drawing for explaining an active oxygen measuring structure according to an embodiment of the present application. FIG. 6 is a diagram illustrating the cell affinity of an active oxygen measuring structure according to an embodiment of the present application. FIG. 7 is a drawing for explaining a method for manufacturing an active oxygen measuring device according to an embodiment of the present application. FIG. 8 is a drawing for explaining the step of forming a functional layer according to an embodiment of the present application. FIG. 9 is a drawing for explaining the step of forming a sensing layer according to an embodiment of the present application. FIG. 10 is a drawing for e