Functional molded body and production method of the same

Abstract

To provide a functional molded body capable of capturing virus, heavy metal, or the like.SOLUTION: A production method of a functional molded body includes steps of: (A) generating a radical on a surface layer of a substrate by irradiating the substrate with electron beams; (B) reacting the radical on the surface layer of the substrate with a polymer brush monomer having an epoxy group, and subjecting a polymer brush to secondary polymerization to dispose the polymer brush on the surface layer of the substrate; and (C) introducing, to the polymer brush, a functional structure to be a host structure of a host-guest reaction using an introduction substance by reacting the epoxy group of the polymer brush with the introduction substance. The host-guest reaction is any one selected from the group consisting of antibody-antigen reaction, ligand-receptor reaction, and chelate-complex reaction.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a functional molded article and a manufacturing method thereof, and in particular to a functional molded article having a polymer brush layer and having functional groups provided on the polymer brush layer, which is suitable for capturing or concentrating various target substances by the functional groups. [Background technology]

[0002] The recent outbreak of COVID-19 has raised interest in viruses, and sewage surveillance projects to predict the spread of infectious diseases through sewage monitoring are being implemented in Japan and other countries. However, this technique requires concentration of sewage because the amount of coronavirus detected in the feces of infected individuals is low, and there are many issues, such as the time lag between collection and detection and the establishment of operating methods. Virus concentration techniques include (a) the antibody method and (b) the negative charge membrane concentration method.

[0003] (a) The antibody method includes an active adsorption method in which an antibody that specifically recognizes the target virus is adsorbed onto the surface of a substrate, and a passive adsorption method in which the antibody is directly bound to the substrate. The former active adsorption method is driven mainly by electrostatic interactions, and although preparation is easy, there are problems with peeling of the adsorbed antibody and shielding of the antigen-binding site due to structural denaturation. The latter passive adsorption method is a method in which a substrate having a carboxyl group or an amino group is brought into an intermediate activation state using a bivalent reagent to form a covalent bond with the target antibody, and does not have the problems of the active adsorption method described above. However, since it is a monolayer adsorption onto the substrate surface, there is a problem that the antigen concentration rate is low.

[0004] (b) The negative charge membrane concentration method involves preparing a membrane that has a positive charge to match the negative charge of the virus, and adsorbing the virus through electrostatic interaction. This method can also be applied to resins, and is a very simple method, but it has the problem of low concentration efficiency because it also adsorbs negatively charged substances other than viruses (e.g. proteins).

[0005] Furthermore, in recent years, rapid industrial development in developing countries has led to the emission of heavy metals through the production of non-ferrous metals and the burning of fossil fuels. Heavy metals thus emitted into the environment are polluting water, a resource essential to life. It has been reported that rapid industrial development has caused thick smog, and has seriously affected the environment, polluting the air, water, soil, and agricultural products with heavy metals. In other words, heavy metal pollution occurs not only in water, but also in soil through the movement of air and rain.

[0006] Mercury, for example, is a harmful pollutant regulated by the U.S. Environmental Protection Agency (EPA) and is known to cause brain and nerve disorders due to its toxicity based on bioaccumulation and non-biodegradability. Another threat of mercury pollution is the health damage caused by mercury methylation. Against this background, the Mercury Export Ban Act was enacted to reduce the use of mercury for commercial purposes worldwide. In other words, technology that can remove mercury contained in wastewater is needed. Chemical precipitation, ion adsorption, membrane filtration, and other wastewater treatment technologies have been applied, but they have problems such as low capacity and low selectivity.

[0007] In addition, expensive equipment is used to analyze hazardous metals such as mercury, but there is a growing need for simple on-site measurement. However, there is a problem with measuring with these simple testers, etc., in that the concentrations of the target metals are low, making on-site measurement difficult. Therefore, it is necessary to develop pretreatment materials that can recover and concentrate specific hazardous metals.

[0008] Methods used to concentrate mercury include (c) activated carbon, (d) solvent extraction, (e) ion exchange resins, and (f) chelating resins.

[0009] (c) While activated carbon has a high adsorption capacity for mercury, it has no selectivity, and there are problems with the inhibition of concentration by coexisting substances and with the separation operation after adsorption. (d) The solvent extraction method uses an extractant that has a sulfur-based functional group that has a high affinity for mercury. Although it has high selectivity, it has problems such as the need to use a large amount of organic solvent and a separation procedure. (e) The driving force for concentration using ion exchange resins is electrostatic interaction, and positively charged substances other than mercury are also adsorbed, resulting in a problem of extremely low selectivity. (f) Chelating resins have sulfur-based functional groups that have a high affinity for mercury fixed inside the resin, and have excellent mercury selectivity. However, because the functional groups exist in a single layer on the pore surface of the resin, there is a problem that a large amount of resin is required to increase the adsorption capacity.

[0010] Meanwhile, it has also been considered to provide various physical properties by providing a so-called polymer brush or the like by graft polymerization on the surface of a substrate. For example, Patent Document 1 discloses a method for producing an anion selective adsorptive porous membrane having neutral hydroxyl groups and anion exchange groups, which is characterized in that a substrate membrane made of a polyolefin or a copolymer of an olefin and a halogenated olefin is irradiated with ionizing radiation, and then glycidyl methacrylate or glycidyl acrylate is grafted in a gas phase, and then, if necessary, epoxy groups are partially ring-opened with an acidic liquid, and then ammonia or an organic amine is added.

[0011] Patent Document 2 discloses a method for producing a selective adsorptive porous membrane, in which a porous substrate membrane having a substantially three-dimensional network structure, which is made of polyolefin, a copolymer of olefin and halogenated olefin, or polyvinylidene fluoride, is irradiated with ionizing radiation, and then a vinyl monomer having an epoxy group is graft-polymerized, and then the resulting mixture is reacted with alcohols, phenols, or monoamines under alkaline conditions, and the remaining epoxy groups are converted into diols. [Prior art documents] [Patent documents]

[0012] [Patent Document 1] Japanese Patent Application Publication No. 2-132132 [Patent Document 2] Japanese Patent Application Publication No. 8-141392 Summary of the Invention [Problem to be solved by the invention]

[0013] As mentioned above, there are various situations where it is required to capture viruses, heavy metals, and the like.

[0014] An object of the present invention is to provide a functional molded article capable of capturing viruses, heavy metals, and the like. [Means for solving the problem]

[0015] The present inventors have conducted extensive research to solve the above problems, and as a result have found that the following invention meets the above object, thereby completing the present invention.

[0016] <1> A step (A) of irradiating a substrate with an electron beam to generate radicals on a surface layer of the substrate; a step (B) of reacting a monomer for polymer brushes having an epoxy group with the radicals of the surface layer of the substrate to secondary polymerize the polymer brushes, thereby providing the polymer brushes on the surface layer of the substrate; and (C) a step of reacting a substance to be introduced with the epoxy group of the polymer brush to introduce a functional structure to the polymer brush, the functional structure being a host structure for a host-guest reaction using the substance to be introduced, The method for producing a functional molded article, wherein the host-guest reaction is any one selected from the group consisting of an antibody-antigen reaction, a ligand-receptor reaction, and a chelate-complex reaction. <2> The polymer brush monomer is glycidyl methacrylate or glycidyl acrylate. <1> The manufacturing method described in <3> The substance to be introduced in the step (C) is a diamine compound, a protein crosslinking agent, and a functional substance, The step (C) of introducing includes a step (C-1) of reacting the epoxy group of the polymer brush with a diamine compound to bond an amino group derived from the diamine compound; a step (C-2) of reacting the protein crosslinking agent that reacts with the amino group; A step (C-3) of reacting the functional substance with the protein crosslinking agent is carried out. <2> The manufacturing method described in <4> The protein crosslinking agent is a bifunctional reagent and / or a dialdehyde compound, and the functional substance is a functional protein. <3> The manufacturing method described in <5> The bifunctional reagent is N-(11-maleimidoundecanoyloxy)sulfosuccinimide, sodium salt, and the dialdehyde compound is glutaraldehyde. <4> The manufacturing method described in <6> The substance to be introduced is a sulfur-containing amine compound having a structure containing an amine and a sulfur atom, and the host structure has a nitrogen atom and a sulfur atom derived from the sulfur-containing amine compound. <2> The manufacturing method described in <7> The structure containing a sulfur atom in the sulfur-containing amine compound is any one selected from the group consisting of thiol, sulfide, and dithiocarboxylic acid. <6> The manufacturing method described in <8> The sulfur-containing amine compound is one or more substances selected from the group consisting of 4-imidazoledithiocarboxylic acid, 3-methylthiopropylamine, 2-aminoethanethiol, and 2,2'-thiobisethylamine. <7> The manufacturing method described in <9> A substrate, a polymer brush layer provided on a surface layer of the substrate, and a functional structure that serves as a host structure for a host-guest reaction and is introduced into the polymer brush layer, The host-guest reaction is any one selected from the group consisting of an antibody-antigen reaction, a ligand-receptor reaction, and a chelate-complex reaction. Effect of the Invention

[0017] The functional molded article of the present invention can capture viruses, heavy metals, and the like. [Brief description of the drawings]

[0018] [Figure 1] FIG. 2 is a flow diagram of an example of a method for producing a functional molded article of the present invention. [Diagram 2] FIG. 1 is a schematic diagram showing a flow of a method for producing a functional molded article according to the present invention. [Diagram 3] FIG. 1 is a schematic diagram of a test device used in the examples. [Figure 4] 1 is an image showing the results of a luminescence test carried out in an example. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] The following is a detailed description of the embodiments of the present invention, but the following description of the constituent elements is one example (representative example) of the embodiment of the present invention, and the present invention is not limited to the following content as long as the gist of the present invention is not changed. Note that in this specification, when the expression "~" is used, it is used as an expression including the numerical values ​​before and after it.

[0020] [Method of manufacturing the functional molded article of the present invention] The method for producing a functional molded article of the present invention includes the steps of: (A) irradiating a substrate with an electron beam to generate radicals on the surface layer of the substrate; (B) reacting a monomer for polymer brush having an epoxy group with the radicals on the surface layer of the substrate to secondary polymerize the polymer brush and provide the polymer brush on the surface layer of the substrate; and (C) reacting a substance for introduction with the epoxy group of the polymer brush to introduce a functional structure to the polymer brush, the functional structure being a host structure for a host-guest reaction using the substance for introduction, the host-guest reaction being any one selected from the group consisting of an antibody-antigen reaction, a ligand-receptor reaction, and a chelate-complex reaction. In the present application, the method for producing a functional molded article of the present invention may be simply referred to as the method for production of the present invention.

[0021] [Functional Molded Article of the Present Invention] The functional molded article of the present invention has a substrate, a polymer brush layer provided on the surface layer of the substrate, and a functional structure that serves as a host structure for a host-guest reaction introduced into the polymer brush layer, wherein the host-guest reaction is any one selected from the group consisting of an antibody-antigen reaction, a ligand-receptor reaction, and a chelate-complex reaction.

[0022] In the present application, the functional molded article of the present invention can be obtained by the method for producing a functional molded article of the present invention, and the corresponding configurations in the present application can be mutually utilized.

[0023] Electron beam graft polymerization is a method in which a polymeric material (substrate) is irradiated with an electron beam to dissociate the carbon-hydrogen (CH) bond, and then vinyl monomer is brought into contact with the generated radical to cause addition polymerization. The C=C bond in the reacted vinyl monomer is broken, one side bonds with the substrate, and the other side becomes a radical and becomes a new reaction point.

[0024] In this way, polymer chains, or graft chains, grow on the substrate through addition polymerization, and new functional groups can be introduced by chemically modifying these chains. In other words, the idea was that by introducing molecules containing sulfur atoms, it would be possible to develop a filter that exhibits mercury adsorption properties.

[0025] The inventors have focused on mercury as a heavy metal and developed filters that can recover mercury efficiently and quickly by densely immobilizing molecules containing sulfur atoms, which have a high affinity for mercury, onto a hollow fiber membrane. Based on this knowledge, they have obtained a functional membrane that can capture heavy metals.

[0026] Furthermore, the inventors developed a functional material that can capture low concentrations of viruses with high sensitivity and high yield by introducing a bivalent reagent into a flat membrane and immobilizing antibodies to the target virus at a high density, and based on this knowledge, they obtained a functional membrane that can capture viruses. The present invention is based on these findings.

[0027] FIG. 1 is a flow diagram of an example of a method for producing a functional molded article of the present invention. Step S11 is a process for generating radicals on the surface layer of the substrate. Step S21 is a process of providing a polymer brush on the surface layer by utilizing radicals on the surface layer. Step S31 is a process for introducing functional groups into the polymer brush, thereby obtaining a functional molded article having functional groups on the surface layer of the molded article serving as the substrate.

[0028] Fig. 2 is a schematic diagram showing the flow of the method for producing a functional molded article of the present invention. Here, an electron beam is irradiated onto a film-like substrate made of polyethylene. This generates radicals on the surface layer. A reactive monomer is secondarily polymerized with these radicals to obtain a polymer brush. By introducing functional groups into this polymer brush, a functional film in which functional groups are densely arranged can be obtained.

[0029] [Step (A) of generating radicals] The radical generating step (A) is a step in which radicals are generated on the surface layer of the substrate. A resin having a CH structure is used as the substrate. By irradiating this CH structure with an electron beam, H atoms are separated and C· radicals are generated on the surface layer of the substrate. The degree of radical generation is appropriately set according to the shape and physical properties of the substrate, the degree to which the polymer brush is desired to be provided, etc. The degree of radicalization is set based on the irradiation dose of the electron beam, etc.

[0030] [Base material] The functional molded body of the present invention is a molded body having a functional group introduced thereinto as a substrate. The substrate can be a molded body that can generate radicals by electron beam irradiation and can maintain any shape under the conditions of use. More specifically, the substrate is a molded body of a resin having a CH structure.

[0031] The material of the substrate may be, for example, polyolefin resins such as polyethylene and polypropylene; polystyrene resins such as polystyrene, acrylonitrile-styrene copolymer (AS resin), and acrylonitrile-butadiene-styrene copolymer (ABS resin); polyvinyl chloride; polyvinylidene chloride; acrylic resins; polyamide; polycarbonate; polyallyl ester; polyimide; polyacetate; polyether ketone, polyether sulfone; polyphenylene oxide, polyphenylene sulfide; cellulose, etc.

[0032] In particular, it is preferable to use a material that is versatile, easily available, easy to process, and suitable for generating radicals and secondary polymerization of polymer brushes, etc. As such, polyethylene is particularly suitable.

[0033] The shape of the substrate may be any molded product, including planar ones such as membranes, sheets, and plates, three-dimensional ones such as hollow fiber membranes, blocks, cylinders, tubes, and containers, and fibers, suitable for the intended use of the guest substance to be captured.

[0034] Representative substrates suitable for use in the production method of the present invention include polyethylene membranes, films, hollow fiber membranes, and the like.

[0035] [Electron beam irradiation] In the manufacturing method of the present invention, the substrate is irradiated with an electron beam to generate radicals. The means for irradiating the electron beam is not particularly limited, and is appropriately selected according to the shape of the substrate and the degree to which the surface layer of the substrate is desired to be radicalized. The electron beam irradiation can be based on the absorbed dose, and can be, for example, about 10 kGy to 1,000 kGy, about 50 kGy to 500 kGy, or about 100 kGy to 300 kGy.

[0036] [Step (B) of Providing a Polymer Brush] The step (B) of providing a polymer brush is a step of providing a polymer brush by secondary polymerization of a monomer for polymer brush using radicals on the surface layer of the base material after the step (A).

[0037] [Monomers for polymer brushes] The monomer for the polymer brush is a monomer for forming a polymer brush. The monomer for the polymer brush is one that leaves an epoxy group when the polymer brush is formed by secondary polymerization.

[0038] As the monomer for the polymer brush, for example, a vinyl monomer having an epoxy group can be used. In this case, the vinyl group undergoes secondary polymerization to form a polymer brush, and this polymer brush has an epoxy group.

[0039] Specific examples of monomers for polymer brushes include glycidyl methacrylate (GMA), glycidyl acrylate, etc. In particular, glycidyl methacrylate is suitable for the production method of the present invention because it forms a polymer brush and has an epoxy group.

[0040] [Secondary Polymerization] The secondary polymerization is a reaction in which a monomer for polymer brushes is polymerized with radicals on the surface layer of a substrate to form a polymer brush. By contacting the substrate in which radicals have been generated with the monomer for polymer brushes, the radicals on the substrate react with the monomer for polymer brushes to cause secondary polymerization, and a polymer brush can be formed on the surface layer of the substrate.

[0041] The means for secondary polymerization can be appropriately set according to the shape of the substrate, the degree and state of the radicals, the type of monomer for the polymer brush, etc. In the secondary polymerization, it is possible to appropriately adjust the fluidity of the monomer for the polymer brush so as to increase the opportunities for the monomer for the polymer brush to come into contact with the radicals, adjust the peripheral conditions to prevent side reactions or inhibitory reactions, add energy to promote the reaction, and so on.

[0042] For example, from the viewpoint of fluidity, the monomer for polymer brushes can be used in the secondary polymerization as a mixed solution mixed with a solvent as appropriate. From the viewpoint of ambient conditions, the secondary polymerization can be performed under nitrogen replacement or reduced pressure to reduce oxygen or air. Ultrasonic irradiation, heating, shaking, etc. can also be performed as appropriate.

[0043] The reaction stops when the monomer for the polymer brush is used up in the secondary polymerization. The conditions until the reaction stops can be controlled by time, but the reaction can also be stopped by changing the conditions to make the reaction less likely by washing or by grasping the concentration of the monomer for the polymer brush.

[0044] [Polymer brush] The functional molded article of the present invention has a polymer brush on the surface layer of a substrate. The polymer brush is made of a polymer obtained by polymerizing a monomer for the polymer brush, and one end of the main chain of the polymer brush reacts with a radical of the substrate and bonds to the substrate. The increase in the dry weight of the polymer brush before and after secondary polymerization can be used as an indicator of graft polymerization.

[0045] [Step (C) of introducing functional groups] The step (C) of introducing a functional group is a step, which is carried out after the step (B), of introducing a functional structure, which serves as a host structure in a host-guest reaction, into the polymer brush.

[0046] [Host-guest reaction] The host-guest reaction may be any one selected from the group consisting of an antibody-antigen reaction, a ligand-receptor reaction, and a chelate-complex reaction.

[0047] [Introduce host structure] The introduction of the functional structure that will become the host structure is carried out using an introduction substance. The introduction substance is a substance having a structure that reacts with the epoxy group of the polymer brush. The introduction substance may itself have a functional structure that will become the host structure. Alternatively, as the introduction substance, a bonding compound for reacting with a substance having a functional structure that will become the host structure and a compound having a functional structure that will become the host structure may be used, and these may be reacted in sequence to introduce the functional structure.

[0048] [Introduction of host structures using protein crosslinkers] A diamine compound, a protein crosslinking agent, and a functional substance are used as the substances to be introduced in the introducing step (C), and the introducing step (C) can include a step (C-1) of reacting the epoxy group of the polymer brush with a diamine compound to bond an amino group derived from the diamine compound, a step (C-2) of reacting the protein crosslinking agent which reacts with the amino group, and a step (C-3) of reacting the functional substance which reacts with the protein crosslinking agent.

[0049] In step (C-1), the epoxy groups of the polymer brush are first reacted with a diamine compound such as ethylenediamine, and the amino groups (amine, -NH2) introduced into the polymer brush are then available for reaction with a protein crosslinker in step (C-2).

[0050] [Diamine compounds] A diamine compound is a compound having two amino groups in the molecule. Formula (1) is a generalized structural formula of an example of a diamine compound. Formula (1) has two amino groups (-NH2). R1 ​​in formula (1) is a structure that bonds each amine. R1 is, for example, an alkylene group having 1 to 12 carbon atoms (-C n H 2n R1 is preferably an alkylene group having 1 to 8 carbon atoms, and particularly preferably an alkylene group having 2 to 6 carbon atoms. As the diamine compound, for example, ethylenediamine, etc. can be used.

[0051] [ka]

[0052] [Protein crosslinking agent] Protein crosslinking agents are used to crosslink protein structures. In particular, those that bind to amine structures contained in proteins are used. In the present invention, the protein crosslinking agent is reacted with the amine structure introduced into the primer brush using a diamine compound, and one side of the protein crosslinking agent is bound to the amine structure.

[0053] [Bifunctional Reagents] Bifunctional reagents can be used as protein crosslinkers. Bifunctional reagents have two functional groups and are widely used in immunoassays and protein research. Bifunctional reagents can be compounds with "N-hydroxysuccinimide active esters" (NHS esters), "N-hydroxysuccinimide active esters containing sulfonic acid" (Sulfo-NHS esters), and "maleimide groups". NHS esters and Sulfo-NHS esters are sometimes called NHS esters because they share a common part of their structure. This bifunctional reagent reacts with the amine structure (-NH2) introduced into the polymer brush and the NHS ester or maleimide group to extend the polymer brush. The NHS ester or maleimide group on the other end binds to proteins such as primary antibodies.

[0054] Formula (2-1) and formula (2-2) are generalized structural formulas of examples of bifunctional reagents. Formula (2-1) has a maleimide group and an NHS ester. Formula (2-2) has a maleimide group and a sulfo-NHS ester. Formula (2-1) and formula (2-2) combine different structures, but both may have an NHS ester or the like. In formula (2-1) and formula (2-2), R 21 , R 22 is a structure that connects each structure.

[0055] R21 And, R 22 are each independently, for example, an alkylene group having 1 to 20 carbon atoms (-C n H 2n -) can be used. R 21 Or R 22 is preferably an alkylene group having 4 to 14 carbon atoms, and particularly preferably an alkylene group having 8 to 12 carbon atoms.

[0056] [ka]

[0057] As such a bifunctional reagent, for example, bifunctional reagents such as "Hetero-bifunctional Reagents" and "Homo-bifunctional Reagents" manufactured by Dojindo Chemical Industries, Ltd. can be used.

[0058] "Hetero-bifunctional Reagents" are a group of reagents that have an NHS ester (OSu) group that reacts with primary amino groups, and a maleimide group that reacts with thiols (SH groups). "Homo-bifunctional Reagents" are a group of reagents that have two NHS esters. These can be used to bind two proteins that are close to each other, or to make compounds that have amino groups amino-reactive. In particular, hetero-bifunctional reagents are preferred, as they are expected to react selectively, with one structure binding to the structure on the polymer brush side and the other binding to a functional protein.

[0059] Examples of bifunctional reagents that can be used include "N-(6-Maleimidocaproyloxy)succinimide", "N-(4-Maleimidobutyryloxy)succinimide", "N-(6-Maleimidocaproyloxy)sulfosuccinimide, sodium salt", "N-(4-Maleimidobutyryloxy)sulfosuccinimide, sodium salt", "N-(8-Maleimidocapryloxy)sulfosuccinimide, sodium salt", "N-(11-Maleimidoundecanoyloxy)sulfosuccinimide, sodium salt", and "N-[(4-Maleimidomethyl)cyclohexylcarbonyloxy]sulfosuccinimide, sodium salt". In particular, "N-(11-Maleimidoundecanoyloxy)sulfosuccinimide, sodium salt" (Sulfo-KMUS) is preferred, as this type of alkyl group is long and the epoxy group is located away from the polymer brush, making it less likely to cause steric hindrance and easier to attach functional substances.

[0060] In addition, examples of bifunctional reagents that can be used include "Bis(sulfosuccinimidyl)suberate, disodium salt", "Dithiobis(succinimidyl undecanoate)", "Dithiobis(succinimidyl hexanoate)", "Dithiobis(succinimidyl propionate)", and "Dithiobis(sulfosuccinimidyl propionate), disodium salt".

[0061] [Dialdehyde compounds] A dialdehyde compound can be used as the protein crosslinking agent. A dialdehyde compound is a compound having two aldehyde groups in the molecule. Formula (3) is a generalized structural formula of an example of a dialdehyde compound. The structure of formula (3) has two aldehydes. R2 in formula (3) is a structure that bonds each aldehyde. R3 is, for example, an alkylene group having 1 to 12 carbon atoms (-C n H 2n -). R3 is preferably an alkylene group having 1 to 8 carbon atoms, and particularly preferably an alkylene group having 2 to 6 carbon atoms. As the dialdehyde compound, for example, glutaraldehyde (1,5-pentanedial) and the like can be used.

[0062] [ka]

[0063] In the present invention, a functional protein can be bound to a polymer brush using a protein crosslinking agent. Examples of such functional proteins include protein G, protein A, protein L, lectin, and antibody. These functional proteins can be appropriately modified for use, for example, by adding an antibody or by capturing a sugar chain.

[0064] When a bifunctional reagent is used as the protein crosslinking agent, a functional substance that reacts with the bifunctional reagent and has a host structure is further used, such as a substance having an antibody structure or a substance having a receptor structure.

[0065] [antibody] The antibody may be, for example, an IgG antibody or an immunoglobulin such as IgM, IgA, IgD, or IgE. Furthermore, a secondary antibody or the like may be bound to the antibody.

[0066] [Receptor structure] It is also possible to introduce a receptor structure for receptor-ligand reactions. The ligand can be a low molecular weight substrate for an enzyme or a hormone. The receptor structure is a structure that corresponds to these ligands. For example, biotin, avidin, etc. can be used.

[0067] [Measurement of antibody fixation] A molecule having an amino group or a thiol group is reacted with the epoxy group, and then a bivalent reagent is immobilized. An antibody-immobilized membrane can be produced by directly binding an antibody to this bivalent reagent. In the present invention, the antibody is directly bound to a high-concentration polymer brush extending three-dimensionally from a substrate via a bivalent reagent, and therefore the amount of antibody per unit area is high, resulting in a higher amount of virus capture than conventional materials with a monolayer of antibody immobilized thereon.

[0068] Since the antibody can be immobilized on the substrate in a three-dimensional layer at a high concentration, the antigen concentration rate per unit area is higher than when the antibody is immobilized in a monolayer. In addition, since the antibody is directly bonded to the polymer brush via a bivalent reagent (crosslinker), denaturation due to electric charge and peeling of the antibody, which occur in active adsorption, are less likely to occur. The functional molded article of the present invention can be used, for example, in immunochromatography, PCR, sewer surveillance, etc., to improve the efficiency of virus concentration.

[0069] Since it can achieve a concentration efficiency far higher than that of existing virus concentration materials, it is expected to improve the sensitivity of immunochromatography and PCR methods. When applied to sewerage surveillance, it may become a new concentration material for COVID-19, which has not yet been established. In addition, since multiple antibodies can be immobilized at once on one type of substrate, it is expected to be applied to multiplex analysis of viruses. The technology of the present invention can be applied not only to antibodies and chelating reagents, but also to enzymes, ionic liquids, etc.

[0070] By introducing a bivalent reagent into the polymer brush and then binding the antibody of the virus to be concentrated to the bivalent reagent, it is possible to take advantage of the high antigen recognition ability of the antibody to produce a material that can efficiently concentrate only specific viruses.

[0071] [Introduction of host structures using sulfur-containing amine compounds] The functional molded article of the present invention can have a substance to be introduced that is a sulfur-containing amine-based compound having a structure containing an amine and a sulfur atom, and a host structure that has a nitrogen atom and a sulfur atom derived from the sulfur-containing amine-based compound.

[0072] The structure containing a sulfur atom in the sulfur-containing amine compound may be any one selected from the group consisting of thiol, sulfide, and dithioate, and the structure of the amine compound is a primary amine or a secondary amine.

[0073] [Thiol] Thiol is a structure in which the oxygen atom of a hydroxyl group is replaced with a sulfur atom. Formula (4) is a generalized structural formula of a sulfur-containing amine compound having a thiol structure. Formula (4) has a thiol structure. R4 in formula (4) is a structure in which an amino group and SH are bonded together. R4 is, for example, an alkylene group having 1 to 12 carbon atoms (-C n H 2n -). R4 is preferably an alkylene group having 1 to 8 carbon atoms, and particularly preferably an alkylene group having 2 to 6 carbon atoms. As this compound, for example, 2-aminoethanethiol can be used.

[0074] [ka]

[0075] [Sulfide] A sulfide is a structure in which divalent sulfur is substituted with two organic groups. Formula (5) is a generalized structural formula of a sulfur-containing amine compound having a sulfide structure. The compound of formula (5) has a sulfide structure. R in formula (5)51 is one of the organic groups. 51 For example, an alkyl group having 1 to 12 carbon atoms (-C n H 2n+1 ) and alkylene groups (-C n H 2n One end of R - can be an amino group. 51 R in formula (5) preferably has an alkyl group or alkylene group having 1 to 8 carbon atoms, and particularly preferably has an alkyl group or alkylene group having 2 to 6 carbon atoms. 52 is a structure that bonds an amino group and SH. R 52 For example, an alkylene group having 1 to 12 carbon atoms (-C n H 2n -) can be used. R 52 is preferably an alkylene group having 1 to 8 carbon atoms, and particularly preferably an alkylene group having 2 to 6 carbon atoms. As this compound, for example, 3-methylthiopropylamine (MTPA), 2,2'-thiobisethylamine (TBEA), etc. can be used.

[0076] [ka]

[0077] The dithiocarboxylic acid may be, for example, 4-imidazoledithiocarboxylic acid (IDTCA).

[0078] [Sulfur-containing amine compounds] As such a sulfur-containing amine compound, one or more substances selected from the group consisting of 4-imidazoledithiocarboxylic acid (IDTCA), 3-methylthiopropylamine (MTPA), 2-aminoethanethiol (AET), and 2,2'-thiobisethylamine (TBEA) can be used.

[0079] As an example of a method using a chelate-complex reaction, the following can be used: A chelate molecule having a sulfur atom in the molecule is fixed between the epoxies of a GMA polymer that becomes a polymer brush. Compared to conventional chelate resins in which chelate molecules are arranged in a single layer, the functional molded article of the present invention has chelate molecules fixed in a high density state to a three-dimensionally extended high-concentration polymer brush, resulting in a high adsorption capacity for metals such as mercury per unit area.

[0080] In other words, because sulfur-based molecules, which have a high affinity for mercury, can be immobilized in a three-dimensional layer at high concentration, rather than in a monolayer on the substrate, the adsorption capacity is higher than that of existing adsorbents, and the concentration efficiency is higher. In addition, the heavy metals that form complexes are captured by the chelate, so it is less susceptible to the effects of coexisting substances.

[0081] The functional molded article of the present invention can recover metals such as mercury at high efficiency and high speed by immobilizing molecules having sulfur atoms, which have a high affinity for metals such as mercury, on a substrate at high density. This is suitable as a pretreatment material for recovering or concentrating specific hazardous metals.

[0082] In this way, the functional molded article of the present invention is produced by irradiating a substrate such as a general-purpose resin with an electron beam to generate radicals, and then contacting the substrate with a reactive monomer to produce a polymer brush. By introducing an appropriate functional group into the polymer brush, new functionality can be imparted, and the molded article can be used as a molded article that captures various guest substances by utilizing a guest-host reaction.

[0083] [Applications of functional molded bodies] The functional molded article of the present invention can be used to capture guest substances in accordance with the host structure. The capture of guest substances can be performed by any method that is suitable for the shape of the substrate and the conditions in which the guest substances are present. For example, a fluid such as a liquid containing a guest substance is brought into contact with the functional molded body, and the guest substance in the fluid is captured by the host structure of the functional molded body. This contact may be continuous by flowing the fluid, or the functional molded body may be immersed in the fluid. The guest substance captured by the host structure may be appropriately desorbed and collected. EXAMPLES

[0084] The present invention will be described in more detail below with reference to examples. However, the present invention is not limited to the following examples unless the gist of the present invention is changed.

[0085] [Example 1] Preparation of functional membrane for mercury capture A functional molded article having a chelate structure was produced using a sulfur-containing amine compound.

[0086] Hollow fiber membrane: As a porous hollow fiber membrane made of polyethylene (PE), a polymer substrate, Asahi Kasei Chemical Corporation's Microza MF (outer diameter 3 mm, inner diameter 2 mm, pore size 0.3 μm) was used.

[0087] Mercury standard solution (Hg 1000) was from Wako Pure Chemical Industries, Ltd. N,N-Dimethylformamide (DMF): Wako Pure Chemical Industries, Ltd. Glycidyl methacrylate (GMA): Wako Pure Chemical Industries, Ltd. Ethylenediamine (EDA): Wako Pure Chemical Industries, Ltd. Ethanol (EtOH): Wako Pure Chemical Industries, Ltd. Methanol (MeOH): Wako Pure Chemical Industries, Ltd. Ammonia water (NH3): Wako Pure Chemical Industries, Ltd. Hydrochloric acid (HCl): Wako Pure Chemical Industries, Ltd. Nitric acid (HNO3): Wako Pure Chemical Industries, Ltd. 2-Aminoethanethiol (AET): First-class or special-grade reagent from Tokyo Chemical Industry Co., Ltd. 2,2'-Thiobisethylamine (TBEA): First-class or special-grade reagent from Tokyo Chemical Industry Co., Ltd. 3-Methylthiopropylamine (MTPA): First-class or special-grade reagent from Tokyo Chemical Industry Co., Ltd. 4-Imidazoledithiocarboxylic acid (IDTCA): Sigma-Aldrich Co. LLC, Santa Cruz Biotechnology, Inc.

[0088] [Electron beam graft polymerization] 1) At the Kyushu EB Center of HV Corporation, a porous hollow fiber membrane made of PE was irradiated with a total of 200 kGy of electron beams to generate radicals. 2) Nitrogen was bubbled through a 10 vol% GMA / MeOH mixed solution for more than 1 hour to replace the oxygen in the solution with nitrogen. 3) The membrane in which radicals were generated was placed in a vacuumed reaction tube, and then a mixed solution of 10 vol% GMA / MeOH was poured in to immerse the membrane. Next, the membrane was irradiated with 40 kHz ultrasonic waves for 1 minute in an ultrasonic cleaner (MCS-6, AS ONE Corporation) (US CLEANER, AS ONE Corporation) to remove the air from inside the membrane, and then the graft polymerization reaction was carried out while shaking for 10 minutes in a 40°C constant temperature shaking water bath (T-N225, Thomas Scientific Instruments Co., Ltd.). 4) The membrane was then washed twice alternately with DMF and MeOH, rinsed twice with pure water, placed in an ultrasonic cleaner while immersed in pure water, and dried for 24 hours in a dryer (DNS-600D, Tokyo Rikakikai Co., Ltd.) at 80°C.

[0089] The graft ratio was calculated as the amount of introduced GMA using formula (1). dg[%] = (W1-W0) / W0 × 100 (1) Here, W0 and W1 are the dry weights of the membrane before and after graft polymerization. In this study, a GMA membrane with a graft ratio of about 70% was prepared.

[0090] [Introduction of functional groups] The structures of the sulfur-containing molecule and the nitrogen-containing molecule for comparison that were introduced into the membrane as functional groups are shown in the following formulas: EDA was used as a comparative example that does not contain sulfur atoms.

[0091] [ka]

[0092] AET, TBEA, and MTPA were adjusted to 1.5 mol / L using pure water. About 5 g of IDTCA was dissolved in methanol, filtered, and then concentrated to 25 mL using a rotary evaporator (N-1000V-W, EYELA Tokyo Rikakikai Co., Ltd.). The introduction of functional groups was carried out by measuring the weight of the dried GMA membrane, immersing the GMA membrane once in EtOH and three times in pure water, and cleaning it in an ultrasonic cleaner at 25° C. for 15 minutes. The washed membrane was immersed in the prepared functional group solution and reacted in a windowed thermostatic water bath (TBN302DA, Advantech Co., Ltd.) set at 80°C to introduce functional groups into the epoxy groups of the GMA. The IDTCA solution was reacted at 60° C. because the solvent was MeOH. After the functional groups were introduced, the membrane was immersed in pure water, EtOH, and pure water in that order, washed in an ultrasonic cleaner, and then dried in a dryer. The IDTCA membrane was immersed in MeOH twice and in pure water three times, washed in an ultrasonic cleaner, and then dried in a dryer. The functional group introduction rate was calculated from the change in membrane weight after drying before and after the introduction of functional groups using the following formula.

[0093] Introduction rate (%) = "difference in membrane weight before and after functional group introduction (g) / GMA membrane weight" x 100

[0094] [Mercury adsorption / desorption test] An arbitrary amount of ethanol was passed through each of the prepared functional group-introduced membranes using a permeation apparatus as outlined in Figure 3, and then approximately 10 mL of 0.1 mol / L hydrochloric acid was passed through the membranes to open the unreacted epoxy groups. Pure water was then passed through the membrane to wash the inside, and then a mercury solution of pH 1.0 adjusted to an arbitrary concentration was passed through the membrane at 2.5 mL / min to adsorb mercury to the functional group-introduced membrane. After mercury adsorption was saturated, 20 mL of pure water was passed through the membrane, and 25 mL of ammonia water adjusted to pH 10 was passed through the membrane to desorb mercury from the membrane. The mercury concentration was calculated by measuring the collected solution with a microwave plasma atomic emission spectrometer (4100MP-AES, Agilent Technologies, Inc.: MP-AES).

[0095] The amount of mercury adsorption was calculated using the following formula. Mercury adsorption amount (mg / g) = "Amount of adsorbed mercury (mg) / membrane weight (g)"

[0096] The mercury desorption rate was calculated using the following formula. Mercury desorption rate (%) = "amount of desorbed mercury (mg) / amount of adsorbed mercury (mg)" x 100

[0097] [Results and Discussion] Mercury adsorption / desorption test Adsorption breakthrough curves were obtained when 5 ppm and 10 ppm mercury solutions were passed through the PE membrane and EDA membrane (77% introduction rate). The horizontal axis of the adsorption breakthrough curve is the amount of feed solution relative to the membrane volume, and the vertical axis is the effluent concentration relative to the initial concentration. The amount of mercury adsorbed was 0.31 mg / g for the PE membrane and 0.40 mg / g for the EDA membrane, meaning that almost no mercury was adsorbed by either membrane.

[0098] Next, the results were obtained when a 10 ppm mercury solution was passed through an AET membrane with a loading rate of 63%. The amount of mercury adsorbed was 11.45 mg / g, and the desorption rate was 16.9%. Mercury was not adsorbed by the PE membrane and EDA membrane, which do not contain sulfur atoms, but was adsorbed by the AET membrane, which does contain sulfur atoms, confirming that there is an affinity between sulfur atoms and mercury even within the membrane.

[0099] Furthermore, to compare the difference between thiol groups and sulfides among molecules containing sulfur atoms, a comparison was made between TBEA membranes and MTPA membranes. Results obtained when a mercury solution was passed through an MTPA membrane with a loading rate of 68% were obtained when a 100 ppm mercury solution was passed through the membrane.

[0100] The mercury adsorption amount of the TBEA film was 6.69mg / g, and the desorption rate was 18.8%, while the mercury adsorption amount of the MTPA film was 86.17mg / g, and the desorption rate was 69.8%, so the MTPA film showed about 13 times the mercury adsorption amount of the TBEA film. This is because the TBEA film has amino groups at both ends, which bridge the polymer brushes, and the frequency factor with mercury may have decreased due to the reduced degree of freedom of the polymer brushes, and the polymer brushes may not be able to take a conformation to form a complex with mercury due to the reduced degree of freedom. In addition, the TBEA film was about half the introduction rate and the amount of adsorption of the AET film, so if the introduction rates of the AET and TBEA films are the same, the amount of mercury adsorption is thought to be about the same.

[0101] It was expected that the amount of mercury adsorbed would be high when thiol groups with high affinity for mercury are present in the membrane, as in the case of the AET membrane, but in fact the result was the same as that of TBEA. Therefore, it was considered that the thiol groups may have reacted nucleophilically with the epoxy groups, so the bonding mode with the epoxy groups was confirmed using a maleimide solution that reacts specifically with thiol groups. When the AET membrane was immersed in the maleimide solution, no maleimide was introduced, suggesting that in the AET membrane, both the thiol groups and amino groups reacted with the epoxy groups and are likely to crosslink the polymer brushes. Therefore, it is considered that the adsorption amounts of the AET membrane and the TBEA membrane were the same.

[0102] Next, we obtained the results when a 200 ppm mercury solution was passed through an IDTCA membrane with a loading rate of 51%. The amount of mercury adsorbed was 66.46 mg / g, and the desorption rate was 28.6%. In addition, when the loading rate was 66%, the amount of mercury adsorbed was 134.1 mg / g, but no breakthrough occurred.

[0103] The reason for the high adsorption amount of the IDTCA film is that the nucleophilicity of dithioates is reduced due to the delocalization of electrons, and the thiol groups do not bond with the epoxy groups, and only the imidazole groups react nucleophilically with the epoxy groups. Therefore, the degree of freedom of the polymer brush does not decrease, and the frequency factor with mercury does not decrease, which is thought to be the reason for the high adsorption amount. In addition, since it has two sulfur atoms, it is thought to have a high ability to form complexes with mercury. However, while the adsorption amount is high, the desorption rate is less than half that of MTPA. This is thought to be because the bond between the sulfur atoms and mercury is strong due to the presence of two sulfur atoms.

[0104] Next, the results were obtained when a 100 ppm mercury solution was passed through an IDTCA membrane with an introduction rate of 33%. The amount of mercury adsorbed was 10.72 mg / g, and the desorption rate was 44.9%, which was 1.6 times higher than that of a membrane with an introduction rate of 51%. This is thought to be because the lower the introduction rate of IDTCA, the lower the local density of IDTCA in the membrane, making it impossible to take a conformation to form a complex with mercury, resulting in a decrease in the amount of adsorption and an improvement in the amount of desorption due to the difficulty in forming a stable complex.

[0105] In comparing thiol groups and sulfides, it was found that sulfides showed higher adsorption and desorption rates than the AET and MTPA membranes. However, when the functional groups bridged the polymer brushes, both the adsorption amount and desorption rate became similar to those of thiols. The results obtained so far are shown in Table 1, which shows the adsorption / desorption test results for various functional membranes.

[0106] [Table 1]

[0107] This paper summarizes the main results of developing a filter capable of concentrating mercury with high efficiency by introducing molecules containing sulfur atoms into the membrane and selecting functional groups suitable for concentrating mercury. (1) To concentrate mercury, a functional group that has a sulfur atom in the molecule and reacts with an epoxy ring only at one end is suitable. (2) When mercury is adsorbed, it is present in large amounts on the inside of the membrane, and when it is desorbed, it moves to the outside and is desorbed after repeated adsorption and desorption.

[0108] [Example 2] Production of functional membrane for capturing viruses Using a bivalent reagent and an IgG antibody, we produced a membrane-like functional molded product that can capture viruses, etc.

[0109] A polyethylene flat membrane was used instead of the hollow fiber membrane in accordance with Example 1. Glycidyl methacrylate (GMA) was graft-polymerized onto the polyethylene flat membrane irradiated with an electron beam. The shaking time for this graft polymerization was 20 minutes, and the graft ratio of GMA was 200%. Then, ethylenediamine (EDA) was introduced to obtain an EDA membrane. Moreover, 2-aminoethanethiol (AET) was introduced instead of EDA to prepare an AET membrane.

[0110] Next, the EDA membrane was immersed in a 2 mmol / L aqueous solution of N-(11-Maleimidoundecanoyloxy) sulfosuccinimide (Sulfo-KMUS) in an ice bath to attempt the introduction of a bifunctional reagent.

[0111] In addition, we attempted to introduce a bifunctional reagent into the AET membrane by immersing it in a 2 mmol / L DMSO solution of Sulfo-KMUS.

[0112] [Results / Discussion] The introduction of functional groups into the GMA membrane was calculated to be 56% for the EDA membrane and 46% for the AET membrane. The introduction rate of the bifunctional reagent into each membrane was 39% for the EDA membrane relative to the weight of the added bifunctional reagent, showing a high introduction rate of the bifunctional reagent. This is expected to lead to the realization of a virus capture material. On the other hand, although the introduction of functional groups into the AET membrane was confirmed, the introduction of the bifunctional reagent did not progress to the point of being introduced.

[0113] [Luminescence test] Based on this test, we verified the immobilization of IgG using HRP-anti-IgG, which specifically recognizes the immobilized IgG. Figure 4 shows the results of an exposure photograph.

[0114] The top of Figure 4 shows the state at the time of exposure photography, and identifies the location of the membrane. The bottom of Figure 4 shows photographs of luminescence. (a) is an EDA membrane (membrane up to the point where EDA was introduced). (b) is a bivalent reagent membrane (membrane up to the point where bivalent reagent was introduced). (c) to (e) are membranes in which the amount of IgG liquid was changed and IgG was introduced after the bivalent reagent. In (c) to (e), luminescence was confirmed, and it was confirmed that the secondary antibody could be fixed, as the antibody was successfully introduced.

[0115] [Example 3] Preparation of Protein G Immobilized Membrane A protein G immobilized membrane was prepared. [reagent] Glutaraldehyde: Fujifilm Wako Pure Chemical Industries, Ltd. "25% Glutaraldehyde Solution Wako Grade 1" Protein G: Fujifilm Wako Pure Chemical Industries, Ltd. "Protein G, recombinant" · IgG antibody: Fujifilm Wako Pure Chemical Industries, Ltd. "Normal rabbit IgG, whole molecule, purified product"

[0116] [Production method] 1) Graft polymerization of GMA Electron beam graft polymerization was carried out in the same manner as in Example 2 to graft polymerize glycidyl methacrylate (GMA).

[0117] 2) Ethylenediamine introduction Similarly, in a manner similar to that of Example 2, ethylenediamine was introduced into a graft polymer of glycidyl methacrylate (GMA) to obtain an EDA membrane.

[0118] 3) Glutaraldehyde introduction Glutaraldehyde was used as a cross-linking agent to bind protein G to amino groups. The EDA membrane was immersed in a 0.5% by volume aqueous solution of glutaraldehyde (GA) at 25°C (room temperature) for about one day, and then washed with pure water several times.

[0119] 4) Protein G adsorption Protein G specifically recognizes and captures IgG antibodies. A 0.1g / L protein G solution was passed through a cylindrical porous hollow fiber membrane using a batch-type permeation device. The flowing protein G solution was sampled and the amount of protein G adsorbed to the membrane was calculated by spectrophotometry. This was continued until the absorbance no longer changed, that is, until the adsorption equilibrium was reached. As a result, the amount of protein G immobilized was 28 (g-protein G / kg-fiber).

[0120] 5) IgG antibody adsorption A 0.2g / L IgG antibody solution was run through a batch-type permeation device. The IgG solution running through was sampled at regular intervals, and the IgG adsorption behavior to the membrane was evaluated by spectrophotometry. The absorbance decreased over time, confirming that adsorption was progressing, and adsorption reached saturation in about 60 minutes, capturing the IgG.

[0121] [Example 4] Preparation of lectin-immobilized membrane [reagent] Lectin: MP Biomedicals, Inc. "Concanavalin A" Glycoprotein: Fujifilm Wako Pure Chemical Industries, Ltd. "Albumin, egg-derived"

[0122] A lectin-immobilized membrane was prepared in accordance with Example 3. Instead of protein G in Example 3, a lectin (concanavalin A) was immobilized.

[0123] Lectins recognize and capture glycans bound to proteins. The flowing lectin solution was sampled at regular intervals, and the lectin adsorption behavior to the membrane was evaluated by spectrophotometry. In the membrane prepared in this example, the lectin (concanavalin A) captured the glycoprotein (albumin).

[0124] It was found that the protein G fixed membrane recognized and adsorbed IgG antibodies, while the lectin (concanavalin A was used in this case) fixed membrane recognized and adsorbed glycoproteins (ovalbumin was used in this case). [Industrial Applicability]

[0125] The functional molded article of the present invention can be utilized for adsorbing or capturing viruses, mercury, etc., and is therefore industrially useful.

Claims

1. A step (A) of irradiating a substrate with an electron beam to generate radicals on a surface layer of the substrate; a step (B) of reacting a monomer for polymer brushes having an epoxy group with the radicals of the surface layer of the substrate to secondary polymerize the polymer brushes, thereby providing the polymer brushes on the surface layer of the substrate; and (C) a step of reacting a substance to be introduced with the epoxy group of the polymer brush to introduce a functional structure to the polymer brush, the functional structure being a host structure for a host-guest reaction using the substance to be introduced, The method for producing a functional molded article, wherein the host-guest reaction is any one selected from the group consisting of an antibody-antigen reaction, a ligand-receptor reaction, and a chelate-complex reaction.

2. The method according to claim 1 , wherein the monomer for the polymer brush is glycidyl methacrylate or glycidyl acrylate.

3. The substance to be introduced in the introducing step (C) is a diamine compound, a protein crosslinking agent, and a functional substance, The step (C) of introducing includes a step (C-1) of reacting the epoxy group of the polymer brush with a diamine compound to bond an amino group derived from the diamine compound; a step (C-2) of reacting the protein crosslinking agent that reacts with the amino group; The method according to claim 2, further comprising the step (C-3) of reacting the functional substance that reacts with the protein crosslinking agent.

4. The method according to claim 3 , wherein the protein crosslinking agent is a bifunctional reagent and / or a dialdehyde compound, and the functional substance is a functional protein.

5. 5. The method according to claim 4, wherein the bifunctional reagent is N-(11-Maleimidoundecanoyloxy)sulfosuccinimide, sodium salt, and the dialdehyde compound is glutaraldehyde.

6. the substance to be introduced is a sulfur-containing amine compound having a structure containing an amine and a sulfur atom, The method according to claim 2 , wherein the host structure has a nitrogen atom and a sulfur atom derived from the sulfur-containing amine compound.

7. The method according to claim 6, wherein the structure containing a sulfur atom in the sulfur-containing amine compound is any one selected from the group consisting of thiol, sulfide, and dithiocarboxylic acid.

8. The method according to claim 7, wherein the sulfur-containing amine compound is one or more substances selected from the group consisting of 4-imidazoledithiocarboxylic acid, 3-methylthiopropylamine, 2-aminoethanethiol, and 2,2'-thiobisethylamine.

9. A substrate, a polymer brush layer provided on a surface layer of the substrate, and a functional structure that serves as a host structure for a host-guest reaction and is introduced into the polymer brush layer, The host-guest reaction is any one selected from the group consisting of an antibody-antigen reaction, a ligand-receptor reaction, and a chelate-complex reaction.

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