Adsorption material and adsorption column

Abstract

[Problem] The purpose of the present invention is to provide an adsorption material and an adsorption column that are capable of reducing adhesion of anything other than a target substance while having high antithrombogenicity. [Solution] The adsorption material of the present invention is characterized by having a water-insoluble carrier having a hydrophilic polymer on the surface thereof, and a ligand fixed to the water-insoluble carrier having a hydrophilic polymer on the surface thereof, capable of interacting with the target substance, and having a weight average molecular weight of 350 to 500,000.

Description

Adsorption materials and adsorption columns

[0001] The present invention relates to an adsorption column that has high antithrombotic properties and reduces adhesion of substances other than target substances.

[0002] Blood purification therapy, in which blood is taken out of the body, disease-causing substances are removed using a blood purifier, and the blood is then returned to the body, is becoming increasingly common. Examples of blood purifiers include artificial kidneys and adsorption columns.

[0003] Artificial kidneys use the principles of dialysis and filtration to remove uremic toxins from patients with renal failure. Adsorption columns use the principle of adsorption to remove specific target substances from patients with drug addiction, hypercholesterolemia, endotoxin-related sepsis, etc. In both blood purifiers, materials that come into contact with blood must have a property known as antithrombogenicity, which makes it difficult for blood components such as platelets to adhere to them.

[0004] For example, it has been reported that patients with chronic renal failure have a high risk of death if their platelet count is low, so there is a need for low platelet adhesion to artificial kidneys. Furthermore, if platelet adhesion to the blood purifier progresses during treatment and leads to blood clots (thrombus), the blood purifier will need to be replaced, which will interrupt treatment and place a heavy burden on patients and medical staff.

[0005] Against this background, many efforts have been made to improve antithrombotic properties. For example, the hollow fiber membranes built into artificial kidneys used in hemodialysis treatment have approximately 10,000 ultrafine channels with inner diameters of approximately 200 μm, making them prone to adhesion of blood components such as platelets and blood coagulation. Attempts have been made to solve these problems by making the hollow fiber membranes hydrophilic.

[0006] For example, in the case of polysulfone hollow fiber membranes, a method has been disclosed in which polyvinylpyrrolidone, a hydrophilic polymer, is mixed into the polysulfone membrane dope at the stage of membrane formation and then molded to impart hydrophilicity to the membrane and inhibit fouling (Patent Document 1).

[0007] However, simply adding a hydrophilic component to a membrane-forming solution does not provide a sufficient adhesion inhibitory effect, and therefore various improvements have been attempted. For example, a method has been disclosed in which the surface of a hollow fiber membrane is modified with a polymer containing an ester group to reduce adhesion of blood components and enhance antithrombotic properties (Patent Documents 2 and 3).

[0008] Furthermore, as a study focusing on the material of the adsorption carrier, a method has been disclosed in which porous particles made of ethylene-vinyl alcohol copolymer and polyvinylpyrrolidone (hereinafter referred to as "PVP"), which are said to be highly biocompatible, are used to suppress adhesion of substances other than the target substance (non-specific adhesion) by utilizing the effect of PVP (Patent Document 4).

[0009] Furthermore, a method has been disclosed in which, in order to suppress nonspecific adhesion to a water-insoluble carrier, a copolymer of a hydrophilic monomer and a hydrophobic monomer is bound to the water-insoluble carrier, and then an antibody is bound to the polymer (Patent Document 5).

[0010] Japanese Patent Publication No. 2-18695 International Publication No. 2009 / 123088 International Publication No. 2018 / 061916 Japanese Patent Application Laid-Open No. 2011-224360 Japanese Patent Application Laid-Open No. 2013-71074

[0011] Patent Documents 1 to 3 are technologies for improving the antithrombogenicity of hollow fiber membranes and the like, but are difficult to apply directly to adsorption materials. That is, adsorption materials require material design that allows them to adsorb target substances, and then they must suppress the adhesion of platelets and the like. Therefore, there is a concern that simply applying antithrombogenic technology to the surface of an adsorption material may reduce the adsorption performance of target substances.

[0012] Patent Document 4 describes that polyvinylpyrrolidone extends some of its molecular chains from the surface of the carrier like whiskers, making the surface hydrophilic, and that a nonspecific adhesion inhibitory effect due to hydrophobic interactions can be expected. However, as described in Patent Documents 2 and 3, the nonspecific adhesion inhibitory effect of PVP is not considered to be high to begin with. In fact, even after extensive investigations by the present inventors, the nonspecific adhesion inhibitory effect of PVP was not sufficient.

[0013] Patent Document 5 is limited to cases where the ligand is an antibody, and describes that control of the orientation of the antibody is important, and is therefore not thought to be applicable to ligands other than antibodies.

[0014] Therefore, an object of the present invention is to provide an adsorption material and an adsorption column that can reduce adhesion of substances other than the target substance, in order to inhibit adhesion of platelets and the like and provide antithrombotic properties.

[0015] As a result of intensive research by the present inventors, the present invention has the following features (1) to (16). (1) An adsorption material comprising a water-insoluble carrier having a hydrophilic polymer on its surface, and a ligand having a weight-average molecular weight of 350 to 500,000, immobilized on the water-insoluble carrier having a hydrophilic polymer on its surface and capable of interacting with a target substance. (2) The adsorption material according to (1), wherein the hydrophilic polymer contains a hydrophilic monomer unit and a hydrophobic monomer unit, and the hydrophobic monomer unit has an ester group. (3) The adsorption material according to (1) or (2), wherein the area percentage of carbon peaks derived from ester groups when the surface is measured by X-ray photoelectron spectroscopy (XPS) is 1 atomic % to 5 atomic %. (4) The adsorption material according to any of (1) to (3), wherein the hydrophilic polymer is electrically neutral. (5) The adsorption material according to any of (1) to (4), wherein the surface has a flexible layer having a thickness of 5 nm to less than 50 nm. (6) The adsorption material according to any one of (1) to (5), wherein the root mean square roughness of the surface in a wet state is 1.0 nm or more and 3.0 nm or less, and the root mean square roughness of the surface in a dry state is 1.5 nm or more and 2.0 nm or less. (7) The adsorption material according to any one of (1) to (6), wherein the weight average molecular weight of the ligand is 500 or more and less than 100,000. (8) The adsorption material according to any one of (1) to (6), wherein the immobilization density of the ligand is 1 nmol / cm ２ 20nmol / cm or more ２The adsorption material according to any one of (1) to (7), which is as follows: (9) The adsorption material according to any one of (1) to (8), which has a shape selected from the group consisting of fibers, beads, films, and hollow fibers. (10) The adsorption material according to any one of (1) to (9), wherein, when the surface is measured by time-of-flight secondary ion mass spectrometry (TOF-SIMS), the ratio of the peak of vinyl propionate to the peak of the functional group derived from the ligand is 0.25 or more and 4.00 or less. (11) The adsorption material according to any one of claims 1 to 10, wherein the ligand contains at least one functional group selected from the group consisting of a carboxyl group, a sulfate group, a sulfonic acid group, a phosphate group, and an amino group. (12) An adsorption column comprising the adsorption material according to claims 1 to 11, and a case having an inlet port for introducing a liquid therein and an outlet port for discharging the liquid therefrom. (13) The adsorption column according to (12), wherein the liquid is at least one liquid selected from the group consisting of blood, plasma, serum, cell culture medium, cell lysate, and purified solutions thereof. (14) A liquid purified using the adsorption column according to (13). (15) A target detection device having the adsorption material according to any one of (1) to (11). (16) The detection device according to (15), wherein the target is at least one target selected from the group consisting of proteins, nucleic acids, peptides, and cells.

[0016] Furthermore, as a result of intensive research by the present inventors, the present invention has the following features (17) to (23). (17) An adsorption material comprising a water-insoluble carrier having ester groups on its surface, and a ligand having a weight-average molecular weight of 500 to 100,000, immobilized on the water-insoluble carrier having ester groups on its surface and capable of interacting with a target substance, wherein the area percentage of carbon peaks derived from the ester groups when the surface is measured by X-ray photoelectron spectroscopy (XPS) is 1 to 5 (atomic %). (18) The adsorption material according to (17), wherein the water-insoluble carrier having ester groups on its surface comprises a charge-neutral, hydrophilic ester group-containing polymer. (19) The adsorption material according to (17) or (18), wherein the surface has a flexible layer having a surface thickness of 5 nm or more and less than 50 nm. (20) The adsorption material according to any one of (17) to (19), wherein the surface has an arithmetic mean roughness of 1.0 μm or less. (21) The adsorption material according to any one of (17) to (20), which has a shape selected from the group consisting of fibers, beads, films, and hollow fibers. (22) An adsorption column comprising the adsorption material according to (17) to (21), and a case having an inlet port for introducing a liquid and an outlet port for discharging the liquid. (23) The adsorption column according to (22), wherein the liquid is selected from the group consisting of blood, plasma, serum, cell culture medium, cell lysate, and purified solutions thereof.

[0017] The adsorption material of the present invention has high antithrombogenicity and can reduce adhesion of substances other than the target substance, and is therefore suitable for use in medical applications requiring antithrombogenicity.

[0018] 1 is a cross-sectional view showing an example of a radial flow type adsorption column, and FIG. 2 is a schematic diagram of a force curve of an atomic force microscope.

[0019] The adsorption material of the present invention is characterized by having a water-insoluble carrier having a hydrophilic polymer on its surface, and a ligand having a weight-average molecular weight of 350 or more and 500,000 or less that is fixed to the water-insoluble carrier having a hydrophilic polymer on its surface and is capable of interacting with a target substance.

[0020] Here, the term "hydrophilic polymer" refers to a polymer that has a high affinity for water, and in the present invention, a polymer that dissolves in an amount of 1 g or more in 100 g of water at 20°C is defined as a hydrophilic polymer.

[0021] The term "ligand" refers to a compound that is capable of adsorbing a target substance by selectively interacting with the target substance.

[0022] Furthermore, "immobilized on a water-insoluble carrier" refers to a state in which the substance remains on the water-insoluble carrier even after immersion in water for 1 hour and washing, and examples thereof include a state in which the substance is coated on the water-insoluble carrier by physical adsorption, a state in which the substance is mixed with the water-insoluble carrier like a polymer alloy, or a state in which the substance is immobilized by chemical bonding.

[0023] The above invention uses a ligand having a weight-average molecular weight of 350 to 500,000 that can interact with a specific target substance, and by immobilizing the ligand on a water-insoluble carrier having a hydrophilic polymer in a specific range on its surface, it is possible to effectively reduce nonspecific adhesion of biological components such as platelets and proteins without impairing the function of the ligand.

[0024] The suppression of adhesion of proteins other than the target substance to the adsorption material, which is not intended for removal, in order to achieve the present invention will be described below.

[0025] Here, "adsorption" is used as a term referring to the active removal of target substances, and "adhesion" refers to the phenomenon in which substances other than the target substances are removed by the adsorption material.

[0026] It is believed that the adhesion of proteins to adsorption materials occurs when the higher-order structure of the protein changes, exposing the internal hydrophobic regions, leading to irreversible adhesion. When proteins adhere or denature, platelet adhesion is induced. Here, there is water around the protein that interacts with the protein and restricts its mobility, known as bound water. It is known that this bound water contributes to the stabilization of the higher-order structure of the protein. In other words, it is thought that when a protein approaches the material and the state of the bound water around the protein is disturbed, the higher-order structure of the protein changes, leading to adhesion.

[0027] As described above, it is known that the prevention of adhesion of proteins other than the target substance can be achieved by incorporating a hydrophilic polymer, which is not too hydrophilic and does not easily disrupt the state of bound water around the protein, into the surface of the water-insoluble carrier.

[0028] Therefore, in the adsorptive material of the present invention, the hydrophilic polymer preferably contains a hydrophilic monomer unit and a hydrophobic monomer unit, and the hydrophobic monomer unit preferably has an ester group.

[0029] Here, the term "monomer unit" refers to a repeating unit in a homopolymer or copolymer obtained by polymerizing a monomer.

[0030] A "hydrophobic monomer unit" is defined as a repeating unit that is poorly soluble or insoluble in water when it is a polymer of itself. Here, "poorly soluble or insoluble in water" means that the solubility in 100 g of pure water at 20°C is 1 g or less. Examples of the hydrophobic monomer unit include a vinyl carboxylate unit, an acrylic ester unit, a methacrylic ester unit, and a styrene derivative unit.

[0031] The carboxylic acid vinyl ester unit is a carboxylic acid vinyl ester structure (—CH(OCO—R)—CH ２ -) (R is a hydrocarbon group in which any hydrogen atom may be substituted with another atom), and examples thereof include vinyl acetate units and vinyl propanoate units.

[0032] The acrylate unit is an acrylate structure (-CH ２ The acrylate ester unit is a monomer unit having the formula —CH(CO—O—R)— (R is a hydrocarbon group, and any hydrogen atom may be substituted with another atom), and examples of the acrylate ester unit include a methyl acrylate unit, an ethyl acrylate unit, and a tert-butyl acrylate unit.

[0033] The methacrylate ester unit is a methacrylate ester structure (-CH ２ -C(CH ３)(CO-O-R)-) (R is a hydrocarbon group, and any hydrogen atom may be substituted with another atom), and examples of the methacrylate ester unit include a methyl methacrylate unit and an isopropyl methacrylate unit.

[0034] The hydrophobic monomer unit preferably has an ester group because it is not too hydrophobic, and more preferably a carboxylic acid ester unit, an acrylic acid ester unit, or a methacrylic acid ester unit. Of these, a carboxylic acid ester unit is preferred because it has little irritation or activation effect on biological components such as blood cells.

[0035] Particularly preferred carboxylic acid ester units in the hydrophobic monomer units are vinyl propanoate units, vinyl butyrate units, vinyl pivalate units, and vinyl pentanoate units, in which the number of carbon atoms in the alkyl group at the side chain terminal is 2 to 7. Particularly preferred acrylic acid ester units are ethyl acrylate units, propyl acrylate units, butyl acrylate units, isobutyl acrylate units, and tert-butyl acrylate units, in which the number of carbon atoms in the alkyl group at the side chain terminal is 2 to 4.

[0036] The methacrylate ester unit in the hydrophobic monomer unit is preferably an ethyl methacrylate unit, a propyl methacrylate unit, a butyl methacrylate unit, an isobutyl methacrylate unit, or a tert-butyl methacrylate unit, in which the number of carbon atoms in the alkyl group at the side chain terminal corresponds to 2 to 7.

[0037] Specific examples of hydrophilic polymers containing an ester group include polyesters, vinyl carboxylic acid esters such as vinyl acetate, acrylic acid esters such as methyl acrylate and methoxyethyl acrylate, and methacrylic acid esters such as methyl methacrylate, ethyl methacrylate, and hydroxyethyl methacrylate.

[0038] On the other hand, a "hydrophilic monomer unit" is defined as a repeating unit that is readily soluble in water in the form of a polymer of itself. Here, "easily soluble in water" means that the solubility in 100 g of pure water at 20°C exceeds 1 g. The hydrophilic monomer unit preferably has a solubility of 10 g or more. Examples of the hydrophilic monomer unit include a vinylpyrrolidone unit, an acrylamide derivative unit, a methacrylamide derivative unit, and a vinylacetamide derivative unit.

[0039] The N-vinylacetamide derivative unit is a vinylacetamide structure (-CH ２ -CH(NR-CO-CH ３ )-) (R is a hydrocarbon group in which any hydrogen atom may be substituted with another atom), and examples of the N-vinylacetamide derivative unit include an N-vinylacetamide unit and an N-methyl-N-vinylacetamide unit.

[0040] The acrylamide derivative unit is an acrylamide structure (-CH ２ —CH(CO—NR １ R ２ )-)(R １ , R ２ are each independently a hydrogen atom or a hydrocarbon group, and any hydrogen atom in the hydrocarbon group may be substituted with another atom), and examples of the acrylamide derivative unit include an acrylamide unit, an N-methylacrylamide unit, an N-isopropylacrylamide unit, and an N-tert-butylacrylamide unit.

[0041] The methacrylamide derivative unit is a compound having a methacrylamide structure (-CH ２ -C(CH ３ ) (CO-NR １ R ２ )-)(R １ , R ２are each independently a hydrogen atom or a hydrocarbon group, and any hydrogen atom in the hydrocarbon group may be substituted with another atom), and examples of the methacrylamide derivative unit include a methacrylamide unit and an N-isopropylmethacrylamide unit.

[0042] Furthermore, the inventors have discovered that in an adsorption material in which a ligand is immobilized on a water-insoluble carrier having a hydrophilic polymer on its surface, the area percentage of the carbon peak derived from ester groups on the surface is set to 1 atomic % or more and 5 atomic % or less, more preferably 2 atomic % or more and 4 atomic % or less, and the use of a ligand with a weight-average molecular weight of 350 to 500,000, more preferably 500 to 100,000, can suppress nonspecific adhesion of substances other than the target substance without affecting the adsorption performance of the ligand and without disrupting the state of bound water around the protein. That is, if the percentage of the carbon peak derived from ester groups is less than 1 atomic %, the effect of suppressing nonspecific adhesion of substances other than the target substance is low, while if it is more than 5 atomic %, the adsorption performance of the ligand is reduced. It has also been found that within this range, the adsorption performance of the ligand is improved. It is speculated that the reason for this is that when nonspecific adhesion occurs on the surface of the water-insoluble carrier, the ligand is physically buried, making it difficult for the target substance to access it.

[0043] The carbon peak derived from the ester group on the surface of the adsorbent material can be obtained by X-ray photoelectron spectroscopy (XPS) measurement, which will be described later.

[0044] Here, we will explain why adhesion can be suppressed without affecting the adsorption performance of the ligand and without disrupting the state of bound water around the protein, even if the ligand has a weight-average molecular weight of 350 or more and 500,000 or less.

[0045] When PVP or polyethylene glycol (hereinafter referred to as "PEG"), which is generally known to have the effect of inhibiting adhesion of biological components such as proteins, is placed on the surface of a water-insoluble carrier, when a ligand is immobilized, a decrease in the adsorption performance of the ligand is observed.

[0046] This is thought to be due to the excluded volume effect caused by the molecular movement of the polymer chains of PVP and PEG in water, in other words, the polymer chains moving like a wiper, which physically prevents the protein from approaching the water-insoluble support, thereby exerting an adhesion-inhibiting effect.However, this wiper effect is also exerted on the target substance adsorbed to the ligand, thereby reducing the adsorption function of the ligand.

[0047] On the other hand, when the amount of ester groups on the surface of the water-insoluble carrier is controlled so that the area percentage of the carbon peak derived from the ester groups is 1 atomic % or more and 5 atomic % or less, adhesion does not occur even upon contact, but it does not mean that physical proximity is not possible. Therefore, the approach of the target substance to the ligand having a weight-average molecular weight of 350 or more and 500,000 or less is not hindered, and it is thought that the adsorption performance of the ligand does not decrease.

[0048] Methods for preparing a water-insoluble carrier having a hydrophilic polymer on its surface include blending a hydrophilic polymer when molding the insoluble carrier, coating the surface of the water-insoluble carrier with a hydrophilic polymer, chemically bonding the surface of the water-insoluble carrier to the hydrophilic polymer, etc. Among these methods, the method of chemically bonding a hydrophilic polymer to the surface of the water-insoluble carrier is particularly preferred from the viewpoint of suppressing elution upon contact with a liquid such as blood.

[0049] Examples of methods for chemical bonding include a method in which a hydrophilic polymer is in contact with a water-insoluble carrier and then irradiated with radiation to generate radicals and form chemical bonds, and a method in which reactive functional groups are introduced into both the water-insoluble carrier and the hydrophilic polymer to form chemical bonds.

[0050] As a method for introducing reactive functional groups into both the water-insoluble carrier and the hydrophilic polymer and chemically bonding them, a method for using a copolymer having a reactive functional group corresponding to the water-insoluble carrier as the hydrophilic polymer can be given.

[0051] Furthermore, in the case of a water-insoluble carrier, when the water-insoluble carrier contains a polyester polymer, examples of the method include a method in which the ester bonds on the surface of the water-insoluble carrier are hydrolyzed by acid or alkali treatment, and the carboxyl groups generated on the surface are chemically bonded to the hydrophilic polymer as reactive groups; a method in which hydroxyl groups, amino groups, etc. introduced by surface treatment with plasma or corona are chemically bonded to the hydrophilic polymer as reactive groups; and a method in which active halogen groups such as halogenated alkyl groups are introduced into the water-insoluble carrier and chemically bonded to the hydrophilic polymer as reactive groups.

[0052] Furthermore, from the viewpoint of the effect of inhibiting non-specific adhesion of proteins and the like, among hydrophilic polymers, electrically neutral hydrophilic polymers and hydrophilic ester group-containing polymers are preferred, and electrically neutral hydrophilic ester group-containing polymers are more preferred.

[0053] In the present invention, the term "charge-neutral polymer" refers collectively to nonionic polymers that do not have functional groups such as amino groups, carboxyl groups, or sulfate groups that dissociate into ions when dissolved in water, and zwitterionic polymers that have functional groups such as carboxybetaine groups, phosphobetaine groups, and sulfobetaine groups, in which cationic groups and anionic groups are present in equal amounts within the molecular skeleton. In the present invention, nonionic polymers are preferred to zwitterionic polymers from the viewpoint of the nonspecific adhesion inhibitory effect.

[0054] In the present invention, as will be described later, polymers that contain 20% or less of monomer units having a charge, such as an amino group or a carboxyl group, relative to the total monomer units, because they are immobilized on a water-insoluble carrier by chemical bonding are also defined as charge-neutral polymers.

[0055] In the present invention, from the viewpoint of adhesion suppression effect, a copolymer containing, as constituent components, a monomer unit A and a monomer unit B represented by the following general formula (I) is preferably used as the hydrophilic polymer. The copolymer may contain one type of monomer unit A and one type of monomer unit B, or two or more types of monomer unit A and monomer unit B. (In the formula, R Ａ represents an alkyl or alkenyl group having 2 to 20 carbon atoms, and RＢ represents a functional group having an amide bond, and X １ , X ２ , X ３ represents a hydrogen atom or a methyl group.)

[0056] R Ａ may be a linear, branched, or cyclic hydrocarbon group present alone, or may contain a hydrocarbon as a constituent element and a functional group such as the above-mentioned carboxybetaine group, phosphobetaine group, or sulfobetaine group.

[0057] Among them, R is the most popular due to its adhesion suppression effect and availability. Ａ is preferably a methyl group, an ethyl group, an n-propyl group, an n-butyl group, an n-pentyl group, an n-hexyl group, an isopropyl group or a tertiary butyl group.

[0058] The monomer unit B is for imparting hydrophilicity to the copolymer, and specifically, N-vinylacetamide, N-vinylpropylamide, N-methylvinylacetamide, vinylpyrrolidone, vinylcaprolactam, acrylamide, N-methylacrylamide, N,N-dimethylacrylamide, amidoalkyl(meth)acrylate, etc. are preferably used. Among them, vinylpyrrolidone or N-vinylacetamide is particularly preferably used from the viewpoint of safety, etc.

[0059] Furthermore, it is preferable to use a copolymer represented by the following general formula (II), in which a monomer unit C having a reactive functional group is added to the above-mentioned monomer unit A and monomer unit B, as the hydrophilic polymer, because the water-insoluble carrier and the hydrophilic polymer can be easily chemically bonded to each other. (In the formula, R Ａ represents an alkyl or alkenyl group having 2 to 20 carbon atoms, and R Ｂrepresents a functional group having an amide bond, Rc represents a functional group selected from the group consisting of an amino group, an azide group, an imino group, a carboxy group, an acid chloride group, an acid anhydride group, an aldehyde group, a hydroxyl group, a phosphate group, a thiol group, an isocyanate group, a thioisocyanate group, an epoxy group, a halogenated alkyl group, a cyano group, a vinyl group, an ethynyl group, a nitro group, a nitroso group, and ionized functional groups thereof, and X １ , X ２ , X ３ represents a hydrogen atom or a methyl group.)

[0060] The functional group possessed by Rc in the monomer unit C is a functional group selected from the group consisting of an amino group, an azide group, an imino group, a carboxy group, an acid chloride group, an acid anhydride group, an aldehyde group, a hydroxyl group, a phosphate group, a thiol group, an isocyanate group, a thioisocyanate group, an epoxy group, a halogenated alkyl group, a cyano group, a vinyl group, an ethynyl group, a nitro group, a nitroso group, and ionized functional groups thereof. From the viewpoint of stability during synthesis of the copolymer, etc., the functional group possessed by Rc in the monomer unit C is preferably an amino group, a carboxy group, an isocyanate group, or an epoxy group.

[0061] Furthermore, when the functional group possessed by Rc is an amino group, the amino group may be either an aliphatic amino group or an aromatic amino group, but an aliphatic amino group is preferred due to its high reactivity. Examples of monomer units having an aliphatic amino group in the side chain include vinylamine, allylamine, alkyleneamine, p-aminoalkylstyrene, and aminoalkyl(meth)acrylate.

[0062] When the functional group possessed by Rc is a carboxy group, the carboxy group may be either an aliphatic carboxy group or an aromatic carboxy group, but an aromatic carboxy group is preferred due to its high reactivity. Examples of the monomer unit having a carboxy group in the side chain include (meth)acrylic acid, 3-butenoic acid, and p-carboxystyrene, and among these, p-carboxystyrene having an aromatic carboxy group is preferred.

[0063] The copolymer is preferably a random copolymer in which the monomer sequence is not biased, rather than a block copolymer in which the monomer sequence is biased. Furthermore, a monomer unit other than the monomer unit A, the monomer unit B, and the monomer unit C may be copolymerized as long as the molar fraction is 30% or less.

[0064] The molar fraction of the monomer unit A is preferably 10 to 90%, more preferably 20 to 75%, and even more preferably 25 to 60%, based on the total constituent monomer units.

[0065] Furthermore, if the molar fraction of the monomer unit B is within an appropriate range, the hydrophilicity of the hydrophilic polymer will be within an appropriate range, thereby enhancing the adhesion suppression effect. Therefore, the molar fraction of the monomer unit B is preferably 10 to 90%, more preferably 30 to 80%, and even more preferably 45 to 70%.

[0066] If the molar fraction of the monomer unit C is within a suitable range, a sufficient amount of copolymer can be immobilized on the surface of the water-insoluble carrier, thereby improving biocompatibility. Therefore, the molar fraction of the monomer unit C is preferably 0.5 to 20%, more preferably 1 to 10%, and even more preferably 1.5 to 5%.

[0067] The molar fraction can be calculated, for example, by performing nuclear magnetic resonance (NMR) measurement and calculating from the ratio of the peak area of ​​a monomer unit to the peak area of ​​all monomer units constituting the copolymer. If the molar fraction cannot be calculated by NMR measurement due to overlapping of peaks or other reasons, the molar fraction may be calculated by elemental analysis.

[0068] The weight-average molecular weight of the hydrophilic polymer is a parameter that affects the thickness of the flexible layer described below, and the weight-average molecular weight is preferably 1,000 to 300,000, more preferably 5,000 to 150,000, and even more preferably 10,000 to 100,000. The weight-average molecular weight of the hydrophilic polymer can be measured by gel permeation chromatography (GPC).

[0069] When a hydrophilic polymer is present on the surface of a water-insoluble carrier, a flexible layer is formed. When a ligand is further immobilized on a water-insoluble carrier having a hydrophilic polymer on its surface, if the flexible layer is within a suitable range, the specific adsorption performance of the ligand can be improved while the non-specific adsorption of the water-insoluble carrier having a hydrophilic polymer on its surface can be further suppressed.

[0070] Therefore, the thickness of the flexible layer is preferably 5 nm or more, more preferably 7 nm or more, and even more preferably 10 nm or more, and is preferably less than 50 nm, more preferably less than 25 nm, and even more preferably less than 15 nm.

[0071] The "thickness of the flexible layer" can be calculated by performing force curve measurements on a water-insoluble carrier in a wet state immersed in water using an atomic force microscope (AFM), as will be described in detail later. The force curve measurement results in a force curve plotted as shown in FIG. 2, with the horizontal axis representing the displacement of the cantilever and the vertical axis representing the force applied to the cantilever. The force curve appears as a line 12 moving parallel to the X-axis until the cantilever probe contacts the surface of the water-insoluble carrier. After the cantilever contacts the surface of the water-insoluble carrier, if a flexible layer is present, the force curve appears as a nonlinear curved portion 13. After passing the curved portion 13, a linear correlation (hereinafter referred to as "straight line 14") is obtained between the displacement of the cantilever and the force applied to the cantilever. The thickness 15 of the flexible layer is defined as the distance from the intersection of the extension of line 12, which runs parallel to the X-axis, and the extension of line 14 on the force curve to the point where the probe tip of the cantilever contacts the surface of the water-insoluble carrier, i.e., the point where bending begins.

[0072] Furthermore, the ligands having a weight-average molecular weight of 350 to 500,000 can be selected from compounds that are optimal for the target substance, and ligands having a weight-average molecular weight of 500 to 100,000 are more preferred.

[0073] Furthermore, the ligand preferably contains at least one functional group selected from the group consisting of a carboxyl group, a sulfate group, a sulfonate group, a phosphate group, and an amino group. Specifically, as long as the weight-average molecular weight is within the above range, it is preferable to use, as the ligand, cationic polymers such as polyethyleneimine, diethylaminoethyldextran, and polyallylamine, anionic polymers such as dextran sulfate, polyacrylic acid, and polyvinyl sulfate, peptides, proteins, nucleic acid aptamers, and polysaccharides.

[0074] For example, when the target substance is endotoxin, polymyxin B can be used. It is also possible to target multiple target substances with one ligand. For example, when polyethyleneimine is used as the ligand, HMGB1 and oxidized low-density apolipoprotein can be adsorbed. Conversely, when selective adsorption of a target substance is desired, it is preferable to use a peptide or a nucleic acid aptamer as the ligand.

[0075] In a water-insoluble carrier having a hydrophilic polymer on its surface, if the weight-average molecular weight of the ligand is too small, it will be easily affected by the ester groups on the surface, resulting in a low specific adsorption ability. Therefore, the weight-average molecular weight of the ligand is preferably 350 or more, more preferably 500 or more, more preferably 1,000 or more, and even more preferably 5,000 or more.

[0076] Furthermore, in a water-insoluble carrier having a hydrophilic polymer on its surface, if the weight-average molecular weight of the ligand is too large, it will have a large effect on the surface, and will cancel out the nonspecific adhesion-inhibiting effect of the hydrophilic polymer, thereby increasing nonspecific adhesion. Therefore, the weight-average molecular weight of the ligand is preferably 500,000 or less, more preferably 100,000 or less, more preferably 50,000 or less, and even more preferably 20,000 or less.

[0077] Furthermore, from the viewpoint of storage stability, when the adsorption material is immersed in water and stored at room temperature (25°C) for one month, the retention rate of the adsorption performance of the ligand for the target substance is preferably 80% or more, and even more preferably 90% or more.

[0078] When the ligand is a peptide, protein, nucleic acid aptamer, etc. with a specific structure and a hydrophilic polymer having an ester group is immobilized on the surface of a water-insoluble carrier, the ester group present on the surface does not disrupt the state of bound water and is unlikely to destabilize the structure of the ligand, resulting in good storage stability.

[0079] From the viewpoint of preventing nonspecific adhesion to the ligand, a strong interaction between the ligand and the target substance is preferable. A method for measuring the interaction between a target substance and a ligand is the isothermal titration calorimetry (ITC) method. In the ITC method, the dissociation constant (Kd), which represents the strength of the interaction, can be calculated from the change in heat quantity when the target substance and the ligand are mixed. The smaller the Kd value, the stronger the binding.

[0080] In the present invention, the Kd of the ligand for the target substance is 10 －５ M or less is preferable, and 10 －７ M or less is more preferable, and 10 －９ It is more preferable that the Kd is small, that is, the interaction between the target substance and the ligand is not too strong to cause a problem, but it is 10 －１４ M or more is preferred.

[0081] The ligand is preferably immobilized by chemical bonding with a water-insoluble carrier having a hydrophilic polymer on its surface. In this case, the ligand and the water-insoluble carrier having a hydrophilic polymer on its surface may be chemically bonded directly or via a spacer. Examples of chemical bonding methods include a method in which an active halogen group contained in the water-insoluble carrier or spacer is reacted with an amino group contained in the ligand to form an amide bond, and a method in which an isocyanate group contained in the water-insoluble carrier or spacer is reacted with a hydroxyl group contained in the ligand to form a urethane bond. However, these bonding methods may be selected appropriately according to the types of ligand, spacer, and water-insoluble carrier.

[0082] Here, the term "spacer" refers to the portion connecting the ligand and the water-insoluble carrier. A short spacer limits the mobility of the ligand, reducing the chance of contact with the target substance and resulting in poor adsorption performance. Therefore, the lower limit of the weight-average molecular weight of the spacer is preferably 50 or more, more preferably 100 or more, and even more preferably 1,000 or more. Furthermore, a long spacer reduces the surface density of the ligand due to the excluded volume effect of the spacer. Therefore, the upper limit of the weight-average molecular weight of the spacer is preferably 50,000 or less, and even more preferably 10,000 or less. Furthermore, to suppress nonspecific adhesion to the spacer, the spacer is preferably a hydrophilic polymer, and more preferably a charge-neutral hydrophilic ester group-containing polymer. In other words, in this case, the spacer is a charge-neutral hydrophilic ester group-containing polymer.

[0083] The water-insoluble carrier refers to a substance that is insoluble in water, and the constituent components may be insoluble in water, and examples thereof include polyaromatic vinyls such as polystyrene, polysulfone-based polymers such as polyethersulfone, polysulfone, and polyarylethersulfone, polyesters such as polyethylene terephthalate and polybutylene terephthalate, polyolefins such as polyethylene, polypropylene, and polymethylpentene, polyacrylonitrile, polymethyl methacrylate, polyurethane, polyetherimide, polyimide, polyamide, copolymers such as styrene / divinylbenzene and ethylene / vinyl alcohol, and naturally occurring polymers such as cellulose. Furthermore, a plurality of these polymers may be used in the water-insoluble carrier.

[0084] The adsorption material, in which a ligand is further fixed to a water-insoluble carrier having an ester group, may be in the form of a fiber, hollow fiber, film, bead, or the like, and can be appropriately used depending on the application, and the water-insoluble carrier may have a porous structure.

[0085] Furthermore, when the liquid to be treated is a liquid containing cells, such as blood or cultured cells, in order to suppress adhesion of blood components such as proteins and cells, it is important that the hydrophilic polymer swells appropriately in water and uniformly coats the surface of the water-insoluble carrier.

[0086] In order to coat the surface of the water-insoluble carrier more uniformly, the lower limit of the root mean square roughness of the surface of the water-insoluble carrier in a wet state is preferably 1.0 nm or more, more preferably 1.1 nm or more, and even more preferably 1.2 nm or more, and the upper limit of the root mean square roughness of the surface of the water-insoluble carrier in a wet state is preferably 3.0 nm or less, more preferably 2.8 nm or less, and even more preferably 2.6 nm or less.

[0087] Here, the term "wet state of the surface of the water-insoluble carrier" is defined as a state in which the water content of the water-insoluble carrier is 65% by mass or more.

[0088] Furthermore, when the surface of the water-insoluble carrier is in a dry state, the coating with the hydrophilic polymer is likely to be non-uniform compared to when it is in a wet state, and the root mean square roughness of the surface of the water-insoluble carrier is likely to be large. Therefore, the upper limit of the root mean square roughness of the surface of the water-insoluble carrier in a dry state is preferably 2.0 nm or less, more preferably 1.9 nm or less, and even more preferably 1.8 nm or less.

[0089] Here, the "dry state of the surface of the water-insoluble carrier" is defined as a state in which the water content of the water-insoluble carrier is 10% by mass or less.

[0090] Here, "root-mean-square roughness" refers to the degree of irregularity measured using an AFM. It is the square root of the average value of the squares of the deviations from the mean line to the roughness curve, and is calculated using the following formula (A). It refers to the root-mean-square roughness (Rq) in accordance with JIS B 0601-2001. In the case of an anisotropic support such as a fiber, the roughness curve is measured in the longitudinal direction of the fiber in a certain direction.

[0091] Furthermore, when the processing liquid is a liquid containing cells, such as blood or cultured cells, the adhesion of blood components such as proteins and cells can be reduced by reducing the surface roughness of the adsorbent material. It is believed that large roughness can cause proteins and cells to accumulate in the recesses and adhere to the material, or that protrusions can physically stimulate cells such as platelets, making them more likely to adhere to the material surface. For these reasons, the upper limit of the arithmetic mean roughness of the surface of the adsorbent material is preferably 1.0 μm or less, more preferably 0.8 μm or less, and even more preferably 0.6 μm or less. There is no particular restriction on the lower limit of the arithmetic mean roughness, and it is sufficient as long as it is 0 μm or more. However, it may be set to 0.1 μm or more in order to increase the surface area and increase the amount of ligand per volume of the adsorbent material.

[0092] Here, the term "arithmetic mean roughness" refers to the value calculated from the following formula (1) when only a reference length L is extracted from a roughness curve in a fixed direction in the direction of the mean line, the x axis is taken in the direction of the mean line of this extracted portion, and the y axis is taken in the direction of the longitudinal magnification, and the roughness curve is expressed as y = f(x), and means the arithmetic mean roughness (Ra) in accordance with JIS B 0601-2001. In the case of an anisotropic support such as a fiber, the roughness curve is measured in the longitudinal direction of the fiber.

[0093] The immobilization density of the ligand in the adsorbent material of the present invention is 1 nmol / cm ２ 20nmol / cm or more ２ The lower limit of the ligand immobilization density is preferably 1 nmol / cm ２ Preferably, 5 nmol / cm or more ２ More preferably, 10 nmol / cm or more ２ On the other hand, the upper limit of the ligand immobilization density is 20 nmol / cm ２ Preferably, 18 nmol / cm or less ２ More preferably, 15 nmol / cm or less ２ The following is even more preferred:

[0094] The "ligand immobilization density" can be calculated by analyzing, by high performance liquid chromatography (HPLC), the ligand immobilization reaction solution to which no adsorbent material has been added, the reaction solution after the ligand immobilization reaction with the addition of the adsorbent material, or the ligand solution extracted from the adsorbent after the ligand immobilization reaction. Specifically, it can be calculated by the method described below in "Calculation of Ligand Immobilization Density."

[0095] Furthermore, when the surface of the adsorbent material of the present invention is measured by time-of-flight secondary ion mass spectrometry (TOF-SIMS), the ratio of the vinyl propionate peak to the peak of the functional group derived from the ligand is 0.25 or more and 4.00 or less. The lower limit of the ratio of the vinyl propionate peak to the peak of the functional group derived from the ligand is preferably 0.25 or more, more preferably 1.00 or more, and even more preferably 2.00 or more. On the other hand, the upper limit of the ratio of the vinyl propionate peak to the peak of the functional group derived from the ligand is preferably 4.00 or less, more preferably 3.50 or less, and even more preferably 3.00 or less.

[0096] The "ligand-derived functional group" refers to a functional group contained only in the ligand immobilized on the water-insoluble support. Details of the measurement by TOF-SIMS will be described later, but since vinyl propionate is released as negative secondary ions when the surface of the water-insoluble support is irradiated with primary ions, the "ligand-derived functional group" is preferably a functional group that is released as a negative secondary ion.

[0097] Furthermore, by incorporating an adsorption material into a case having at least an inlet port for introducing a liquid and an outlet port for discharging the liquid, an adsorption column for selectively removing a target substance can be created.

[0098] The adsorption material to be incorporated into the adsorption column is preferably in the form of fibers, hollow fibers, or beads. From the viewpoints of contact efficiency with the liquid, pressure loss in the adsorption column, ease of manufacture, etc., the diameter of the fibers is preferably 1 μm or more and 100 μm or less, more preferably 2 μm or more and 50 μm or less, and even more preferably 5 μm or more and 30 μm or less.

[0099] The "fiber diameter" can be measured by taking a photograph of a cross section perpendicular to the elongation direction of the fiber at a magnification of 1000 to 3000 times using a scanning electron microscope or the like, and analyzing the obtained image of the fiber cross section. Specifically, it can be measured by the method described below in "Measurement of fiber diameter."

[0100] Furthermore, when the adsorption material is fibrous, it is preferable to use irregular cross-section yarns from the viewpoint of increasing the contact area per fiber volume, and examples of the cross-sectional shapes of irregular cross-section yarns include Y-shaped, I-shaped, and W-shaped cross-sections, polygonal cross-sections such as triangular and square cross-sections, and flower, star, and cloud shapes.

[0101] In the case of a modified cross-section yarn, the diameter of the modified cross-section yarn is the diameter when converted into the area of ​​a circular cross-section. The fiber may be in the form of a knitted fabric, a woven fabric, a nonwoven fabric, or a cut fiber, but a knitted fabric is preferably used because the mesh portion serves as a liquid flow path and the pressure loss in the adsorption column is easily reduced.

[0102] Furthermore, when the adsorption material is beads, the pressure loss in the adsorption column tends to be larger than that of fibers, so the diameter is preferably 100 μm or more and 2 mm or less, more preferably 200 μm or more and 1 mm or less.

[0103] When the adsorption material is a hollow fiber, the inner diameter of the hollow fiber is preferably 100 μm to 2 mm, and more preferably 150 μm to 500 μm. Furthermore, the membrane thickness of the hollow fiber is preferably 20 μm to 500 μm, and more preferably 30 μm to 100 μm.

[0104] The shape of the adsorption column may be either a radial flow type or a tangential flow type, with radial flow being preferred. A radial flow type refers to an adsorption column in which a perpendicular flow exists within the adsorption column relative to the inlet and outlet through which the liquid passes. A tangential flow type refers to an adsorption column in which only a parallel flow exists within the adsorption column relative to the inlet and outlet through which the liquid passes.

[0105] The liquid passed through the adsorption column is preferably selected from blood, plasma, serum, cell culture medium, cell lysate, and purified solutions thereof. Blood is particularly preferred because it effectively suppresses nonspecific adhesion and selectively adsorbs target substances. The preferred use is for blood purification therapy.

[0106] When the adsorption column is used for blood purification treatment, the adsorption material is preferably in the form of fibers, and preferably in the form of a knitted fabric.

[0107] From the viewpoint of maintaining the strength of the material, composite fibers are preferred, and sea-island composite solid fibers are more preferred. Of these, it is preferred to form sea-island composite solid fibers in which the island portions are made of a component that functions as a reinforcing material and the sea portion is made of a component that can fix a ligand or an ester group-containing polymer.

[0108] Examples of island components include polyamide, polyacrylonitrile, polyethylene, polypropylene, nylon, polymethyl methacrylate, and polytetrafluoroethylene. These polymers may be used alone or in combination. Among them, polypropylene is preferred because of its high chemical resistance and excellent thermoplasticity. The sea portion is preferably a polymer that is easy to chemically bond with a ligand or an ester group-containing polymer and is easy to melt-spin. Specifically, polystyrene or a polystyrene derivative is preferably used. Note that polymers other than polystyrene may be alloyed from the viewpoint of fiber strength, etc.

[0109] An example of the internal configuration of an adsorption column of the present invention containing fibers will be described with reference to Figure 1. In Figure 1, reference numeral 1 denotes a container body having an inlet 2 and an outlet 3 at the front and rear ends in the longitudinal direction. A filter 4 and a disk-shaped partition plate 5 are provided inside the inlet 2, and a filter 6 and a disk-shaped partition plate 7 are provided inside the outlet 3.

[0110] Of the two partition plates 5, 7, the front (inlet side) partition plate 5 has an opening 5a in its center, and the rear partition plate 7 has a support protrusion 7a in its center. A number of through-holes 7b are also provided intermittently in the circumferential direction on the outer periphery of the partition plate 7. Furthermore, a pipe 8 is stretched between the opening 5a of the partition plate 5 and the support protrusion 7a of the partition plate 7.

[0111] The pipe 8 has a flow path 9 formed therein for guiding blood, and has a number of through holes 10 in its peripheral wall. The front end of the pipe 8 communicates with the opening 5a of the partition plate 5, and the rear end of the pipe 8 is closed by the support protrusion 7a of the partition plate 7.

[0112] A knitted fabric made of a plurality of fibers 11 is wound around the outer periphery of the pipe 8, forming a plurality of layers. When an adsorption column incorporating these fibers is used for blood purification treatment, a tube forming a circulation circuit between the inlet 2 and the outlet 3 is connected to the blood pool, and blood taken out of the blood pool is supplied to the inlet 2, and harmful substances such as disease-causing proteins, i.e., target adsorbable substances, are adsorbed and removed by the internal fibers 11, and the blood flows out of the outlet 3 and is circulated back into the blood pool.

[0113] Within the adsorption column, blood enters flow path 9 from inlet 2 through filter 4 and then sequentially penetrates fibers 11 through through-holes 10 as it moves through flow path 9. Target adsorbable substances in the blood moving in either radial direction are adsorbed. The blood from which target adsorbable substances have been adsorbed flows out through numerous through-holes 7b on the outer periphery of partition plate 7, passes through filter 6, and then flows out through outlet 3. In the above example, blood flows from opening 5a through flow path 9 in pipe 8 and then flows out through through-hole 10. However, the direction of blood movement in an adsorption column incorporating fibers may be reversed, with blood supplied through outlet 3 and flowing out through inlet 2.

[0114] It is also possible to create target detection devices filled with adsorbent materials.

[0115] The target is preferably at least one selected from the group consisting of a protein, a nucleic acid, a peptide, and a cell.

[0116] The liquid passed through the target detection device is preferably selected from blood, plasma, serum, cell culture medium, cell lysate, and purified solutions thereof. The target can be detected by contacting the target-containing liquid with the adsorbent material, which causes a detectable or measurable color change.

[0117] The present invention will be described below with reference to examples, but the present invention is not limited to these examples.

[0118] An ester group-containing polymer, vinylpyrrolidone / vinyl propanoate / allylamine random copolymer, was chemically bonded to the surface of a sea-island composite fiber made of polystyrene and polypropylene, and polyethyleneimine was chemically bonded as a ligand, to create an adsorptive material in which a ligand was further bonded to a water-insoluble carrier having ester groups on its surface. The nonspecific adhesion inhibitory ability was evaluated using a platelet adhesion test, and the ligand adsorption performance was evaluated using the adsorption performance of HMGB1 or endotoxin. The specific method is described below.

[0119] (1) Preparation of Adsorbent Material (a) Preparation of Knitted Fabric A knitted fabric was prepared from sea-island composite fiber as a water-insoluble carrier. Specifically, a sea-island composite fiber (fiber diameter: 20 μm) was spun. The fiber had 264 island components made of polypropylene J105WT (manufactured by Prime Polymer Co., Ltd.) and a sea component made of 90% by mass of polystyrene (Mw: 181,000) and 10% by mass of polypropylene J105WT (manufactured by Prime Polymer Co., Ltd.), with an island-to-sea ratio (mass ratio) of 50:50. 36 of the resulting fibers were doubled and tubular knitted to obtain a knitted fabric (hereinafter referred to as the "raw knitted fabric").

[0120] Next, reactive functional groups were introduced into the raw knitted fabric in order to chemically bond the hydrophilic polymer using the following procedure. First, 2 g of paraformaldehyde (hereinafter "PFA") was dissolved in a mixed solution of 20 mL of nitrobenzene and 13 mL of sulfuric acid at 10°C (hereinafter "PFA solution"). Furthermore, 47 g of NMCA was dissolved in a mixed solution of 259 mL of nitrobenzene and 169 mL of sulfuric acid at 10°C (hereinafter "NMCA solution"). 10 g of raw knitted fabric was immersed in the PFA solution, and the NMCA solution was quickly added and stirred. After immersion and stirring for 1 hour, the knitted fabric was removed, washed with an excess amount of nitrobenzene, substituted and washed with methanol, and further washed with ion-exchanged water to obtain an α-chloroacetamidomethylated knitted fabric (hereinafter "Intermediate 1"). The entire process from preparing the PFA solution to washing the knitted fabric with methanol was carried out at 15°C or below.

[0121] (b) Synthesis of Copolymer A vinylpyrrolidone / vinyl propanoate / allylamine random copolymer was synthesized by the following procedure as a hydrophilic polymer to be chemically bonded to Intermediate 1. Note that vinyl propanoate is a hydrophobic monomer unit containing an ester group.

[0122] The monomers, polymerization solvent, and polymerization initiator were all manufactured by Tokyo Chemical Industry Co., Ltd. 19.5 g of vinylpyrrolidone, 17.5 g of vinyl propanoate, 1.0 g of allylamine hydrochloride, 56 g of t-amyl alcohol as a polymerization solvent, and 0.175 g of 2,2'-azobis(2,4-dimethylvaleronitrile) as a polymerization initiator were mixed and stirred at 80°C for 6 hours under a nitrogen atmosphere. The reaction solution was cooled to room temperature to terminate the reaction, and then poured into hexane.

[0123] The resulting white precipitate was collected and dried under reduced pressure for 12 hours at 20° C. Thereafter, the copolymer was dissolved in an aqueous solution of sodium bicarbonate (manufactured by Wako Pure Chemical Industries, Ltd.) adjusted to a pH of 8 to remove hydrochloric acid, and water was removed using an evaporator, thereby obtaining the target hydrophilic polymer (copolymer 1).

[0124] １ ​The results of H-NMR measurement showed that the molar fraction of vinylpyrrolidone, vinyl propanoate, and allylamine relative to all constituent monomer units was 72.0%, 25.0%, and 3.0%, respectively.

[0125] The Mw of copolymer 1 calculated from the GPC measurement results shown below was 30,000. First, a 0.1 N lithium nitrate solution containing water and methanol at a volume ratio of 50 / 50 was prepared as a GPC developing solution. 2 mg of copolymer 1 was dissolved in 2 ml of this solution. 100 μL of this solution was injected into a Prominence GPC system (manufactured by Shimadzu Corporation) and measured. The instrument configuration was as follows:

[0126] Pump: LC-20AD, Autosampler: SIL-20AHT, Column oven: CTO-20A, Column: GMPWXL (inner diameter 7.8 mm x 30 cm, particle size 13 μm; manufactured by Tosoh Corporation).

[0127] The flow rate was 0.5 mL / min, and the measurement time was 30 minutes. Detection was performed using a differential refractive index detector RID-10A (Shimadzu Corporation), and the Mw of copolymer 1 was calculated from the peak derived from copolymer 1 that appeared around 15 minutes into the elution time. The molecular weight of copolymer 1 was calculated by rounding off to the nearest hundred. A calibration curve was created using polyethylene oxide standard samples (0.1 kD to 1258 kD; Agilent Technologies).

[0128] A vinylpyrrolidone / allylamine random copolymer was synthesized by the following procedure: A vinylpyrrolidone / allylamine random copolymer (copolymer 2) having a weight average molecular weight of 45,000 was obtained in the same manner as above, except that vinyl propanoate was not added.

[0129] (2b) Modification and Purification of Aptamer In order to immobilize one of the ligands, Anti-HMGB1 aptamer (Aptamer Sciences), to Intermediate 1, the aptamer was modified and purified according to the following procedure.

[0130] Sodium tetraborate was dissolved in a 42.5% aqueous solution of dimethyl sulfoxide to a concentration of 25%. To this solution, the anti-HMGB1 aptamer was added to a concentration of 100 μM, and then dibenzocyclooctyne (DBCO)-amine (Sigma-Aldrich) was mixed to a concentration of 70 μM. This was left to stand at room temperature for 1 hour to obtain the DBCO-bound anti-HMGB1 aptamer (hereinafter referred to as the "modified aptamer").

[0131] To the resulting modified aptamer, 5 M aqueous sodium perchlorate solution in an amount equal to 1 / 10 of the volume of the modified aptamer and acetone in an amount three times the volume of the modified aptamer were added, and the mixture was allowed to stand at -80°C for 1 hour. After standing, the mixture was centrifuged at 4°C and 14,000 rpm for 15 minutes to produce a precipitate, and the supernatant was removed. Next, acetone was added and mixed, and the supernatant was removed. 75% ethanol was added and mixed, and the supernatant was removed. The remaining precipitate was dried at 37°C for 10 minutes, then dissolved in distilled water to obtain a purified aptamer.

[0132] (c) Immobilization of Ligand and Copolymer Polyethyleneimine and anti-HMGB1 aptamer were selected as ligands for adsorbing inflammation-inducing HMGB1, and the cyclic peptide polymyxin B was selected as a ligand for adsorbing endotoxin, a bacterial toxin. These were immobilized to Intermediate 1 together with the copolymer synthesized in (b) above under various conditions to obtain a predetermined adsorption material. Details of the immobilization conditions will be described later.

[0133] (2) X-ray photoelectron spectroscopy (XPS) measurement The surface of each sample (knitted fiber fabric) of the adsorbent material was measured at three points. The measurement samples were rinsed with ultrapure water, dried at room temperature and 0.5 Torr for 10 hours, and then subjected to measurement. The measurement equipment and conditions were as follows:

[0134] Measurement equipment: ESCALAB220iXL Excitation X-ray: monochromatic Al Kα1,2 ray (1486.6 eV) X-ray diameter: 0.15 mm Photoelectron escape angle: 90° (inclination of the detector relative to the sample surface)

[0135] The amount of carbon derived from ester groups was determined by dividing the peaks appearing at +4.0 to +4.2 eV from the main peak (near 285 eV) of C1s CH or C-C, calculating the ratio of the peak area to the total elements, and then determining the amount of carbon derived from ester groups (atomic %). The calculation was rounded to one decimal place.

[0136] (3) Measurement of the flexible layer on the surface of the adsorbent material The surface of the adsorbent material (knitted fiber fabric) was measured using an atomic force microscope (AFM). The measured sample was rinsed with ultrapure water and then dried at room temperature and 0.5 Torr for 10 hours before being used for measurement.

[0137] After attaching the fiber to the sample stage, a water droplet was dropped onto the film to wet it to a moisture content of 65% by mass or more. In this state, a force curve was measured in contact mode (Figure 2). Care was taken to prevent the fiber surface from drying out during the measurement. If a flexible layer was present on the surface when the cantilever approached the fiber, a curved portion 13 was observed between line 12 and line 14 in the force curve obtained by the above measurement. The distance from the intersection of the extension of line 12 and the extension of line 14 to the point where the curvature began was defined as the thickness 15 of the flexible layer. Measurements were performed at 20 randomly selected locations, and the average value was used. The average value was rounded to one decimal place.

[0138] The AFM observation conditions were as follows: scanning probe microscope SPM 9500-J3 (Shimadzu Corporation), observation mode was contact mode, probe was NP-S (120 mm, wide) (Veco Japan), scan range was 5 μm × 5 μm, and scan speed was 1 Hz.

[0139] (4) Measurement of root-mean-square roughness (Rq) The surface of the adsorbent material (knitted fiber fabric) was measured using an atomic force microscope (AFM). The fiber was cut into 5 mm squares, attached to a sample stage, and dried at room temperature and 0.5 Torr for 24 hours to obtain a dried sample. After observing the dried sample under the conditions described below, the sample stage was fixed with transparent nail polish in a glass cell attached to the device and immersed in RO water for 24 hours to obtain a wet sample. The wet sample was also observed in the same manner as the dry sample. By not changing the observation mode, probe, or other scanning conditions, the change between the dry and wet states can be quantitatively evaluated. Measurements were performed at five locations, and the average value was used.

[0140] The AFM observation conditions, including the equipment, observation mode, and probe, were the same as those described in "Measurement of the flexible layer on the surface of the adsorbent material." The scanning range was 2 mm x 2 mm (the z-scale of the image was standardized to 10 nm), and the scanning speed was 0.2 Hz. Image processing was performed by calculating the average value for each x and y line from the obtained raw image, and subtracting this average value from the data to correct the slope. Noise lines were eliminated by eliminating line noise in the image data caused by cantilever needle jumps and instantaneous noise during measurement, based on the inversion of the brightness of the preceding and following lines. In addition, if the image contained regular noise in terms of spatial frequency, image processing was performed using a frequency filter.

[0141] To calculate Rq, the offline function "surface roughness analysis" was used. The analysis range was 1 to 4 μm so as not to pick up specific shapes. ２ (standard is 4 μm ２ The one-dimensional Rq is the square root of the average value of the squares of the deviations from the mean line to the roughness curve in the longitudinal direction of the fiber, and is calculated by the following formula (A).

[0142] (5) Calculation of Ligand Immobilization Density The ligand immobilization density was calculated by analyzing the ligand immobilization reaction solution (hereinafter referred to as the "pre-reaction solution") to which no adsorbent material (knitted fiber fabric) had been added, and the reaction solution after the addition of the knitted fabric and the ligand immobilization reaction (hereinafter referred to as the "post-reaction solution") by HPLC. The post-reaction solution was recovered as follows. The knitted fabric was added to the ligand immobilization reaction solution, and the ligand immobilization reaction was carried out. The entire amount of the ligand immobilization reaction solution was then extracted (hereinafter referred to as "reaction solution A"). Next, the knitted fabric after the ligand immobilization reaction was immersed in the same amount of distilled water as the ligand immobilization reaction solution and washed. The entire amount of the liquid after washing (hereinafter referred to as "reaction solution B") was recovered and mixed with reaction solution A. The knitted fabric was washed again using the same procedure, and the entire amount of the liquid after washing (hereinafter referred to as "reaction solution C") was recovered and mixed with the mixture of reaction solutions A and B. 1 mL was extracted from the mixture of reaction solutions A, B, and C, and filtered through a PTFE membrane filter (pore size 0.45 μm, manufactured by ADVANTEC) to obtain a post-reaction solution for analysis by HPLC. The pre-reaction solution was recovered using the same procedure as the post-reaction solution, except that no knitted fabric was added to the ligand immobilization reaction solution. The pre-reaction solution and the post-reaction solution were each analyzed by HPLC (analysis conditions will be described later), and the ligand immobilization density was calculated from the obtained ligand peak area using the following formula (B): Ligand immobilization density (nmol / cm ２ ) = {(amount of ligand (nmol)) / (knitted fabric area (cm ２ ))} × {1 - (ligand peak area of ​​post-reaction solution) / (ligand peak area of ​​pre-reaction solution)} ... formula (B)

[0143] The HPLC analysis was carried out under the following conditions.

[0144] Measurement device: High-performance liquid chromatograph Prominence (Shimadzu Corporation) Column: X-bridge C18 3.5 μm, 4.6 mm × 150 mm (Waters Corporation) Eluent A: 95 mM triethylammonium acetate / 5% acetonitrile mixture Eluent B: acetonitrile Gradient: 5 to 40% eluent B (20 minutes) Flow rate: 1 mL / min Detection: UV (254 nm)

[0145] (6) Calculation of the ratio of the vinyl propionate peak to the peak of the ligand-derived functional group The surface of the adsorbent material (knitted fiber fabric) was analyzed by TOF-SIMS. The surface of the knitted fabric was dried at room temperature and 0.5 Torr for 24 hours, and then irradiated with primary ions under the conditions described below. ２５ C ２ H － The mass spectrum of ７９ P.O. ３ － The mass spectrum of ７３ C ３ H ５ O ２ － The mass spectrum of the compound was measured. The ratio of the vinyl propionate peak to the peak of the functional group derived from the ligand (hereinafter referred to as "phosphoric acid / vinyl propionate ratio") was calculated by the following formula (C): (phosphoric acid / vinyl propionate ratio) = {( ７９ P.O. ３ － Spectral intensity) / ( ２５ C ２ H － Spectral intensity)} / {( ７３ C ３ H ５ O ２ － Spectral intensity) / ( ２５ C ２ H － (spectral intensity)} ...Equation (C)

[0146] The TOF-SIMS analysis was carried out under the following conditions.

[0147] Measurement device: TOF. SIMS 5 (manufactured by ION-TOF) Primary ion: Bi ３ ＋＋ Primary ion acceleration voltage: 30 kV Pulse width: 21.5 ns Secondary ion polarity: negative Number of scans: 32 scans Measurement range: 300 x 300 μm ２ Mass range (m / z): 0-1500

[0148] (7) Measurement of arithmetic mean roughness The surface of an adsorbent material sample (fiber knitted fabric) dried in a vacuum dryer for 3 hours or more was photographed under the following conditions, and the roughness curve in the longitudinal direction of the fiber was measured.

[0149] Measurement device: Color 3D laser microscope VK-9700 (manufactured by Keyence Corporation) Objective lens magnification: 100x Z-axis direction measurement pitch: 0.1 μm Brightness: 1360 Camera gain: 12 dB

[0150] From the obtained roughness curve, only a portion of a reference length L was cut out in the direction of the mean line, and the x-axis was set in the direction of the mean line of this cut-out portion, and the y-axis was set in the direction of the longitudinal magnification. The roughness curve was expressed as y = f(x), and the arithmetic mean roughness was calculated by the following formula (1) (in accordance with JIS B 0601-2001):

[0151] The reference length L was set to 10 μm, and five locations per sample, i.e., per fiber, were selected as the measurement range. Furthermore, measurement ranges were similarly selected for 10 other samples (10 fibers), and the average value of the values ​​measured in a total of 50 measurement ranges was taken as the arithmetic mean roughness of the surface of the sample. The average values ​​were rounded to the first decimal place.

[0152] (8) Measurement of Fiber Diameter The "fiber diameter" was determined by the following method. 100 fibers were randomly sampled, and a photograph of the cross section (a cross section perpendicular to the elongation direction of the fiber) was taken for each fiber at a magnification of 3000 using a scanning electron microscope. Next, the diameter of each fiber cross section was measured using image analysis software ImageJ. The average value of these values ​​(average diameter of the cross sections of a total of 100 fibers) was calculated, and the value rounded to one decimal place was used as the fiber diameter.

[0153] (9) Platelet Adhesion Test In order to confirm the platelet adhesion inhibitory ability of the surface of the adsorbent material (knitted fiber fabric), the adsorbent material having ester groups on its surface was impregnated into the blood of healthy human volunteers for a predetermined period of time, and the material was then removed and the amount of platelet reduction in the blood before and after impregnation was measured. The measurement method for the knitted fabric is as follows:

[0154] The knitted fabric was cut into a 10 mm diameter disk, and one disk was placed in a polypropylene container. 0.79 mL of healthy human volunteer blood, supplemented with 50 U / mL heparin or 0.13 mg / mL nafamostat mesylate, was added to the container containing the cut knitted fabric disk, and the container was rotated at 6 rpm in a 37°C incubator for 1 hour. The knitted fabric was then removed from the blood, and the platelet concentration in the blood was measured using a hematology analyzer XN-1000V (Sysmex Corporation). The platelet concentration of a blank sample was measured using the same procedure except without the knitted fabric, and the platelet adhesion rate to the knitted fabric was calculated using the following formula (2): Platelet adhesion rate (%) = {(Platelet concentration of blank sample) - (Platelet concentration after contact with knitted fabric)} / (Platelet concentration of blank sample) × 100 (Formula (2))

[0155] (10) HMGB1 Adsorption Test In order to confirm the HMGB1 adsorption ability of polyethyleneimine immobilized on the adsorption material (knitted fiber fabric), an HMGB1 adsorption test was carried out by the following method.

[0156] The knitted fabric was cut into disks with a diameter of 8 mm, and two disks were placed in a polypropylene container and washed with physiological saline. After washing, the knitted fabric was dehydrated and transferred to a new polypropylene container (hereinafter referred to as the "container containing the knitted fabric").

[0157] Next, 0.53 g of bovine serum albumin (hereinafter referred to as "BSA") was added to 15.0 mL of phosphate buffered saline to prepare a 3.5% BSA solution. A 1 mg / mL HMGB1 solution (Shinotest) was serially diluted with this 3.5% BSA solution to obtain a 100 ng / mL HMGB1 solution. 500 μL of the 100 ng / mL HMGB1 solution was added to the knitted fabric container and shaken at 37°C and 500 rpm using a Thermomixer (Eppendorf) for 2 hours.

[0158] After shaking, the HMGB1 solution remaining in the container containing the knitted fabric (hereinafter referred to as the "post-shaking solution") was transferred to a new polypropylene container. HMGB1 in the post-shaking solution was detected using an HMGB1 ELISA Kit Exp (Shinotest Co., Ltd.), and the HMGB1 concentration was quantified by measuring the absorbance at 450 nm using a microplate reader SpectraMax M5 (Molecular Devices Japan Co., Ltd.).

[0159] Furthermore, 500 μL of a 100 ng / mL HMGB1 solution was added to a polypropylene container containing no knitted fabric, and the container was shaken in the same manner as for the container containing the knitted fabric to obtain a blank solution. The HMGB1 concentration of the blank solution was quantified in the same manner as for the solution after shaking, and the HMGB1 adsorption rate of the knitted fabric was calculated using equation (3). HMGB1 adsorption rate (%) = {(HMGB1 concentration of blank solution) - (HMGB1 concentration of solution after shaking)} / (HMGB1 concentration of blank solution) × 100 ... equation (3)

[0160] (11) Endotoxin Adsorption Test In order to confirm the endotoxin adsorption ability of polymyxin B immobilized on a water-insoluble carrier (knitted fiber fabric), an endotoxin adsorption test was carried out by the following method.

[0161] All glassware used in this test was sterilized by dry heat at 250° C. for 2 hours before the test. 1 mg of endotoxin (lipopolysaccharides from Escherichia coli O111:B4; Sigma-Aldrich) was dissolved in 10 mL of distilled water to prepare a 100 μg / mL endotoxin aqueous solution.

[0162] This 100 μg / mL endotoxin aqueous solution was serially diluted with fetal bovine serum (Biosera) to 10 ng / mL, and shaken at 37° C. for 1 hour to obtain an endotoxin adsorption test solution.

[0163] Next, 0.5 g of the knitted fabric was placed in an Erlenmeyer flask and washed with physiological saline, after which fresh physiological saline was added and the mixture was steam sterilized for 30 minutes at 115° C. After steam sterilization, the knitted fabric was washed with distilled water, removed, and transferred to a new Erlenmeyer flask (hereinafter referred to as the "flask containing the knitted fabric").

[0164] 15.0 mL of the endotoxin adsorption test liquid was placed in each of the flasks containing the knitted fabric and the Erlenmeyer flasks not containing the knitted fabric, and the flasks were shaken at 37°C for 2 hours.

[0165] After shaking, 0.1 mL of the LPS adsorption test solution was removed from each of the flasks containing the knitted fabric and the Erlenmeyer flask containing no knitted fabric, transferred to a new polypropylene container, and heated at 70°C for 10 minutes (hereinafter, the endotoxin adsorption test solution removed from the flask containing the knitted fabric will be referred to as the "post-shaking solution," and the endotoxin adsorption test solution from the Erlenmeyer flask containing no knitted fabric will be referred to as the "blank solution").

[0166] After cooling each solution to 15°C, 0.2 mL of each solution was added to a test tube of Limulus HS-J Single Test Wako (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.; hereinafter referred to as "Limulus reagent"). The test tube containing the Limulus reagent to which each solution had been added was placed in a Toxinometer ET-5000 (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) for 50 minutes, and the endotoxin concentration was measured. The endotoxin adsorption rate of the knitted fabric was calculated using formula (4). Endotoxin adsorption rate (%) = {(endotoxin concentration of blank solution) - (endotoxin concentration of solution after shaking)} / (endotoxin concentration of blank solution) x 100 ... formula (4)

[0167] Example 1 10.7 g of polyethyleneimine (hereinafter "PEI", Mw: 10,000) was dissolved in 500 mL of ion-exchanged water, and then aqueous sodium hydroxide solution was added to adjust the pH to 9.5 to prepare a 2.1 mmol / L PEI aqueous solution. 10.0 g of intermediate 1 was immersed in this PEI aqueous solution and stirred for 3 hours. It was then immersed in ion-exchanged water and thoroughly washed to obtain a knitted fabric (hereinafter "intermediate 2") to which PEI was chemically bonded. The entire process from preparing the PEI aqueous solution to washing the knitted fabric with ion-exchanged water was carried out at 40°C. Furthermore, copolymer 1 was dissolved in aqueous sodium hydroxide solution at pH 9.5 to a concentration of 10 μM, and intermediate 2 was immersed and stirred for 3 hours. It was then immersed in ion-exchanged water and thoroughly washed to obtain Example 1, an adsorbent material to which copolymer 1 was also chemically bonded.

[0168] The arithmetic mean roughness of Example 1 was 0.5 μm. Furthermore, as shown in the results in Tables 1 and 3, the ability to adsorb HMGB1, the target of polyethyleneimine, was achieved while suppressing platelet adhesion. Furthermore, after storage in water at room temperature (25° C.) for one month, the retention rate of the platelet adhesion inhibitory effect and the retention rate of HMGB1 adsorption performance were both 90% or higher.

[0169] [Example 2] The same procedure as in Example 1 was carried out, except that the concentration of copolymer 1 was changed to 100 μM, to obtain an adsorption material of Example 2. As shown in the results in Table 1, platelet adhesion was suppressed while the ability to adsorb HMGB1, the target of polyethyleneimine, was exhibited.

[0170] [Example 3] The same procedure as in Example 1 was carried out, except that the concentration of copolymer 1 was changed to 1000 μM, to obtain an adsorption material of Example 3. As shown in the results in Tables 1 and 3, the adsorption ability of HMGB1, the target of polyethyleneimine, was exhibited while suppressing platelet adhesion.

[0171] Example 4 Instead of PEI in Example 2, polymyxin B (Mw: 1,203), a peptide with a cyclic structure, was immobilized on Intermediate 1. That is, 0.65 g of polymyxin B sulfate was dissolved in 1 L of purified water, the pH of which was adjusted to 9.5 with sodium hydroxide, and Intermediate 1 was immersed and stirred for 1 hour. After thorough washing with 0.1 N hydrochloric acid and 10.0 g of purified water, Copolymer 1 was immobilized in the same manner as in Example 2, to obtain the adsorption material of Example 4.

[0172] As shown in the results in Table 1, platelet adhesion was suppressed while the ability to adsorb endotoxin, the target of polymyxin B, was exhibited.

[0173] [Example 5] The same procedure as in Example 1 was carried out, except that the weight-average molecular weight of PEI was changed to 25,000 Mw, to obtain the adsorption material of Example 5. As shown in the results in Table 2, the adsorption ability of HMGB1, the target of PEI, was exhibited while suppressing platelet adhesion.

[0174] [Example 6] N ３PEG-SH was dissolved in a 2 mM aqueous sodium hydroxide solution to a concentration of 20 μM, and the intermediate 1 was immersed and stirred at 60° C. for 1 hour. Thereafter, it was immersed in ion-exchanged water and thoroughly washed. ３ A knitted fabric (hereinafter referred to as "Intermediate 3") was obtained in which copolymer 1 was chemically bonded to PEG-SH. Next, copolymer 1 was dissolved in a sodium hydroxide solution at pH 9.5 to a concentration of 1000 μM, and Intermediate 3 was immersed in the solution and stirred at 40°C for 3 hours. Thereafter, it was immersed in ion-exchanged water and thoroughly washed, and then immersed in N ３ A knitted fabric (hereinafter referred to as "Intermediate 4") was obtained in which PEG-SH and copolymer 1 were chemically bonded. Furthermore, the purified aptamer was dissolved in a 38.5% aqueous acetone solution to a concentration of 25 μM, and Intermediate 4 was immersed and stirred at 25°C for 17 hours. It was then immersed in ion-exchanged water and thoroughly washed, yielding Example 6, an adsorption material also chemically bonded to the purified aptamer.

[0175] As shown in the results in Tables 2 to 4, platelet adhesion was suppressed while the ability to adsorb HMGB1, the target of the anti-HMGB-1 aptamer, was exhibited.

[0176] Example 7 The same procedure as in Example 6 was carried out, except that the concentration of the purified aptamer was changed to 100 μM, to obtain the adsorption material of Example 7. As shown in the results in Table 4, the adsorption ability of HMGB1, the target of the anti-HMGB-1 aptamer, was exhibited while suppressing platelet adhesion.

[0177] Comparative Example 1 The same procedure as in Example 1 was carried out, except that PEI and copolymer 1 were not added to the solution in which intermediate 1 was immersed, to obtain an adsorbent material of Comparative Example 1.

[0178] As shown in the results in Table 1, there was almost no platelet adhesion or adsorption of HMGB1 or endotoxin. The slight adsorption is thought to be due to non-specific interactions.

[0179] Comparative Example 2 The same procedure as in Example 1 was carried out, except that copolymer 1 was not added to the solution in which intermediate 1 was immersed, to obtain an adsorbent material of Comparative Example 2.

[0180] As shown in the results in Tables 1 and 3, although HMGB1 adsorption ability was present, platelets also adhered.

[0181] Comparative Example 3 The same procedure as in Example 2 was carried out, except that PEI was not added to the solution in which the intermediate 1 was immersed, to obtain an adsorbent material of Comparative Example 3.

[0182] As shown in the results in Table 1, platelet adhesion was inhibited, but HMGB1 was hardly adsorbed.

[0183] Comparative Example 4 The same procedure as in Example 2 was carried out, except that the weight average molecular weight of the PEI in which the intermediate 1 was immersed was changed to 750,000 Mw (manufactured by BASF), to obtain an adsorption material of Comparative Example 4.

[0184] As shown in the results in Table 1, HMGB1 was adsorbed, but platelets also adhered.

[0185] Comparative Example 5 The same procedure as in Example 2 was carried out, except that the weight average molecular weight of the PEI in which the intermediate 1 was immersed was changed to 300 Mw (manufactured by Nippon Shokubai Co., Ltd.), to obtain an adsorbent material of Comparative Example 5.

[0186] As shown in the results in Table 1, platelet adhesion was inhibited, but the adsorption performance of HMGB1 was low.

[0187] Comparative Example 6 A PET mesh (single filament diameter: 27 μm, mesh size: 100 μm) used as a water-insoluble carrier was immersed in an aqueous solution of 3.0 wt % potassium permanganate (manufactured by Wako Pure Chemical Industries, Ltd.) and 0.6 mol / L sulfuric acid (manufactured by Wako Pure Chemical Industries, Ltd.), and the PET mesh was hydrolyzed and oxidized by reacting for 3 hours at 60° C. After the reaction, the aqueous solution was removed, and the PET mesh was washed with 6N hydrochloric acid (manufactured by Wako Pure Chemical Industries, Ltd.) and distilled water.

[0188] Next, the PET mesh was immersed in an aqueous solution of 0.5 wt % DMT-MM and 5.0 wt % PEI, and the mixture was allowed to react at 50° C. for 2 hours to chemically bond the PEI to the PET mesh. After the reaction, the aqueous solution was removed, and the mesh was washed with distilled water to obtain the adsorbent material of Comparative Example 6.

[0189] The arithmetic mean roughness of Comparative Example 6 was 0.3 μm. As shown in the results in Table 1, the adsorption rate of HMGB1 was low and the inhibition of platelet adhesion was insufficient.

[0190] Comparative Example 7 The same procedure as in Example 4 was carried out, except that copolymer 1 was changed to copolymer 2, to obtain an adsorbent material of Comparative Example 7.

[0191] As shown in the results in Table 1, the adsorption of endotoxin and the inhibition of platelet adhesion were also insufficient.

[0192] Comparative Example 8 The same procedure as in Example 4 was carried out, except that copolymer 1 was not added, to obtain an adsorbent material of Comparative Example 7.

[0193] As shown in the results in Table 1, endotoxin was adsorbed, but platelets also adhered.

[0194] Comparative Example 9 An adsorption material of Comparative Example 9 was obtained in the same manner as in Example 1, except that the concentration of copolymer 1 was changed to 10,000 μM.

[0195] As shown in the results in Table 3, platelet adhesion was inhibited, but the adsorption performance of HMGB1 was low.

[0196] Comparative Example 10 The same procedure as in Example 6 was carried out, except that the concentration of the purified aptamer was changed to 10 μM, to obtain an adsorption material of Comparative Example 10.

[0197] As shown in the results in Table 4, platelet adhesion was inhibited, but the adsorption performance of HMGB1 was low.

[0198] Comparative Example 11 The same procedure as in Example 6 was carried out, except that the concentration of the purified aptamer was changed to 300 μM, to obtain an adsorption material of Comparative Example 11.

[0199] As shown in the results in Table 4, the adsorption performance of HMGB1 was low, and the inhibition of platelet adhesion was insufficient.

[0200] Comparative Example 12 The same procedure as in Example 6 was carried out, except that the concentration of copolymer 1 was changed to 5 μM, to obtain an adsorbent material of Comparative Example 12.

[0201] As shown in the results in Table 4, HMGB1 was adsorbed, but the inhibition of platelet adhesion was insufficient.

[0202] Comparative Example 13 An adsorption material of Comparative Example 13 was obtained in the same manner as in Example 6, except that the concentration of copolymer 1 was changed to 10,000 μM.

[0203] As shown in the results in Table 4, platelet adhesion was inhibited, but the adsorption performance of HMGB1 was low.

[0204] For Examples 1 to 4 and Comparative Examples 1 to 8, the area percentage of the carbon peak derived from the ester group, the platelet adhesion rate, and the target substance adsorption rate were measured by the above-mentioned methods and are summarized in Table 1.

[0205] For the adsorption materials of Examples 5 and 6, the area percentage of the carbon peak derived from the ester group, the platelet adhesion rate, and the target substance adsorption rate were measured by the above-mentioned methods, and the results are summarized in Table 2.

[0206] For the adsorption materials of Examples 1, 3, and 6 and Comparative Examples 2 and 6, the thickness of the flexible layer, the root mean square roughness in the wet state, the root mean square roughness in the dry state, the platelet adhesion rate, and the target substance adsorption rate were measured using the methods described above, and the results are summarized in Table 3.

[0207] For the adsorption materials of Examples 6 and 7 and Comparative Examples 8 to 12, the ligand immobilization density, phosphate / vinyl propionate ratio, platelet adhesion rate, and target substance adsorption rate were measured using the methods described above, and the results are summarized in Table 4.

[0208]

[0209]

[0210]

[0211]

[0212] By changing the ligand with a weight-average molecular weight of 350 to 500,000 to match the corresponding target substance, the adsorbent material of the present invention and the adsorption column incorporating it can be used to treat patients with acute diseases such as sepsis, diseases such as cancer and autoimmune diseases, and patients in whom changes in blood cell dynamics in the body should be prevented as much as possible, such as the elderly, children, and pregnant women.Furthermore, by changing the corresponding ligand in the same way, the material can also be used in the production of biopharmaceuticals, where only specific substances are selectively removed and the adhesion of cultured cells is suppressed.

[0213] DESCRIPTION OF SYMBOLS 1...container body, 2...inlet, 3...outlet, 4...filter, 5...partition plate, 5a...opening of partition plate, 6...filter, 7...partition plate, 7a...support protrusion of partition plate, 7b...through hole of partition plate, 8...pipe, 9...flow path, 10...through hole, 11...fiber, Q...blood flow, 12...line moving parallel to the X-axis, 13...curved portion, 14...straight line, 15...thickness of flexible layer

Claims

1. An adsorbent material comprising a water-insoluble carrier having a hydrophilic polymer on its surface, and a ligand having a weight average molecular weight of 350 or more and 500,000 or less, which is fixed to the water-insoluble carrier having a hydrophilic polymer on its surface and is capable of interacting with a target substance.

2. The adsorbent material according to claim 1, wherein the hydrophilic polymer contains a hydrophilic monomer unit and a hydrophobic monomer unit, and the hydrophobic monomer unit has an ester group.

3. The adsorbent material according to claim 1 or 2, wherein the area percentage of the carbon peak derived from the ester group when the surface is measured by X-ray photoelectron spectroscopy (XPS) is 1 atomic % or more and 5 atomic % or less.

4. The adsorbent material according to any one of claims 1 to 3, wherein the hydrophilic polymer is electrically neutral.

5. The adsorbent material according to any one of claims 1 to 4, having a flexible layer of 5 nm or more and less than 50 nm on its surface.

6. The adsorbent material according to any one of claims 1 to 5, wherein the root mean square roughness of the surface in the wet state is 1.0 nm or more and 3.0 nm or less, and the root mean square roughness of the surface in the dry state is 1.5 nm or more and 2.0 nm or less.

7. The adsorbent material according to any one of claims 1 to 6, wherein the weight average molecular weight of the ligand is 500 or more and less than 100,000.

8. The immobilization density of the ligand is 1 nmol / cm ２ or more and 20 nmol / cm ２ or less. The adsorbent material according to any one of claims 1 to 7.

9. The adsorbent material according to any one of claims 1 to 8, which is in a shape selected from the group consisting of fibers, beads, films, and hollow fibers.

10. The adsorbent material according to any one of claims 1 to 9, wherein the ratio of the peak of vinyl propionate to the peak of the functional group derived from the ligand when the surface is measured by time-of-flight secondary ion mass spectrometry (TOF-SIMS) is 0.25 or more and 4.00 or less.

11. The adsorbent material according to any one of claims 1 to 10, wherein the ligand contains at least one functional group selected from the group consisting of a carboxyl group, a sulfate group, a sulfonic acid group, a phosphate group, and an amino group.

12. An adsorption column comprising the adsorbent material according to claims 1 to 11, and a case having an inlet port for flowing in a liquid and an outlet port for flowing out the liquid.

13. The adsorption column according to claim 12, wherein the liquid is at least one liquid selected from the group consisting of blood, plasma, serum, cell culture fluid, cell lysate, and purified liquids thereof.

14. A liquid purified using the adsorption column according to claim 13.

15. A target detection device having the adsorbent material according to any one of claims 1 to 11.

16. The detection device according to claim 15, wherein the target is at least one target selected from the group consisting of proteins, nucleic acids, peptides, and cells.

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