Biologically derived component adsorbing material

Abstract

The objective of the present invention is to provide a biologically derived component adsorbing material that exhibits the capability of adsorbing a target substance while non-specific adsorption of proteins is inhibited. The present invention provides a biologically derived component adsorbing material including a water-insoluble carrier having a ligand with an amino group or a negatively charged functional group, a chemical structure containing a halogen atom, and a base material, wherein the ligand and the chemical structure are bonded to the base material, and the halogen atom content is 0.05 mmol to 2.00 mmol per 1 g of the dry weight of the water-insoluble carrier.

Description

Biocomponent adsorption materials

[0001] The present invention relates to a material for adsorbing biological components.

[0002] Blood purification therapy, in which blood is withdrawn from the body, pathogens are removed using a blood purifier, and the blood is then returned to the body, is becoming widespread. Examples of blood purifiers include artificial kidneys and adsorption columns.

[0003] An example of an adsorption column used in blood purification therapy is a blood purification column packed with an adsorption material having a ligand that adsorbs specific target substances in diseases in which an increase in pathogenic substances is observed in the blood, such as sepsis, preeclampsia, and hypercholesterolemia.

[0004] Furthermore, an example of an adsorption column used to purify a target substance from a culture solution of cells, bacteria, viruses, or the like is an affinity separation column capable of specifically adsorbing a target substance.

[0005] Generally, liquids containing biological components, which are the target of treatment using an adsorption column, contain many proteins other than the target substance. Therefore, if the adsorption column adsorbs proteins other than the target substance, there is a concern that the adsorption performance of the target substance will be reduced due to a decrease in the number of adsorption sites for the target substance. Therefore, an adsorption column is required to exhibit the adsorption performance of the target substance while suppressing nonspecific adsorption of proteins.

[0006] Known examples of materials that adsorb target substances include a material in which a functional group having an amino group is introduced onto the surface of a water-insoluble carrier, and which adsorbs high mobility group proteins (Patent Document 1), and a material in which a functional group having an acidic functional group selected from the group consisting of a sulfate group, a sulfite group, and a sulfonic acid group, and an amino group, is introduced, and which simultaneously adsorbs leukocytes and cytokines (Patent Document 2).

[0007] As a material that inhibits nonspecific adsorption of proteins, a protein-adsorption inhibiting material in which a cationic substance is immobilized on a substrate is known (Patent Document 3).

[0008] As a material that can simultaneously exhibit the ability to adsorb target substances and suppress nonspecific adsorption of proteins, an adsorbent for adsorbing IgM antibodies is known, which comprises a water-insoluble carrier to which is attached a ligand having a carboxyl group and a monovalent group consisting of the atomic group remaining after removing one hydrogen atom bonded to a carbon atom constituting a heterocycle from thiophene or a derivative thereof. It has been suggested that the carboxyl group in this adsorbent suppresses adsorption of the protein fibrinogen (Patent Document 4).

[0009] Known biological component adsorption materials containing a water-insoluble carrier have a chemical structure containing halogen atoms and a substrate, and include a water-insoluble carrier to which a ligand is bound, and which has a surface arithmetic mean roughness of 0.01 μm or more and 1.0 μm or less and a surface maximum height roughness of 0.025 μm or more and 10.0 μm or less. These water-insoluble carriers are known to inhibit the adhesion of blood cell components such as platelets (Patent Document 5).

[0010] JP 2012-005827 A International Publication No. 2012 / 057185 JP 2004-201752 A JP 2013-053079 A JP 2024-103469 A

[0011] However, although the materials described in Patent Documents 1 and 2 adsorb target substances by the introduced functional groups, there is no disclosure or suggestion regarding non-specific adsorption to proteins other than the target substances.

[0012] Patent Document 3 shows that a material in which a cationic substance is immobilized on a substrate does not adsorb antibodies, which are proteins, but cationic substances are thought to adsorb negatively charged proteins.

[0013] In the material disclosed in Patent Document 4, the adsorption of fibrinogen is suppressed by the carboxyl group of the ligand, but since carboxyl groups generally interact electrostatically with the amino groups of proteins, it is unclear whether the adsorption of proteins other than fibrinogen is also suppressed.

[0014] The material disclosed in Patent Document 5 is a material in which the arithmetic mean roughness and maximum height roughness of the surface are within a certain range, thereby suppressing adhesion of blood cell components such as platelets, but there is no disclosure or suggestion regarding non-specific adsorption to proteins other than the target substance.

[0015] Therefore, an object of the present invention is to provide a biological component-adsorbing material that exhibits the ability to adsorb target substances while suppressing non-specific adsorption of proteins.

[0016] The present inventors have discovered that in a biological component adsorption material comprising a water-insoluble carrier having a ligand having an amino group or a negatively charged functional group, a chemical structure containing halogen atoms, and a substrate, nonspecific adsorption of proteins can be suppressed by setting the halogen atom content within a predetermined range. That is, the present invention, which solves the above-mentioned problems, includes the following (1) to (7): (1) A biological component adsorption material comprising a water-insoluble carrier having a ligand having an amino group or a negatively charged functional group, a chemical structure containing halogen atoms, and a substrate, wherein the ligand and the chemical structure are bound to the substrate, and the halogen atom content is 0.05 to 2.00 mmol per 1 g of dry weight of the water-insoluble carrier. (2) The biological component adsorption material according to (1), wherein the halogen atom content is 0.06 to 1.50 mmol per 1 g of dry weight of the water-insoluble carrier. (3) The biological component adsorption material according to (1) or (2), wherein the negatively charged functional group is a sulfonic acid group or a carboxyl group. (4) The biological component adsorption material according to any one of (1) to (3), wherein the halogen atom is a chlorine atom. (5) The biological component adsorption material according to any one of (1) to (4), wherein the content of the amino group or negatively charged functional group is 0.04 to 0.50 mmol per 1 g of the dry weight of the water-insoluble carrier. (6) The biological component adsorption material according to any one of (1) to (5), wherein the ligand is a ligand having an ethyleneimine polymer. (7) The biological component adsorption material according to any one of (1) to (5), wherein the negatively charged functional group is a thioalkylsulfonic acid group or a thioalkylcarboxyl group.

[0017] The biological component adsorption material of the present invention exhibits the ability to adsorb target substances while suppressing nonspecific adsorption of proteins, and therefore can be used in blood purification columns for treating diseases in which an increase in pathogenic substances is observed in the blood, affinity separation columns for purifying target substances from liquids containing biological components, and the like.

[0018] The biological component-adsorbing material of the present invention comprises a water-insoluble carrier having a ligand having an amino group or a negatively charged functional group, a chemical structure containing a halogen atom, and a substrate.

[0019] "Adsorption" refers to a state in which a specific substance adheres to a material and does not easily peel off. The principle of adsorption is not particularly limited, but examples include adhesion due to intermolecular forces such as electrostatic interaction, hydrophobic interaction, hydrogen bonding, or van der Waals forces, and physical adhesion such as cell adhesion or phagocytosis by leukocytes.

[0020] "Biologically derived components" refers to substances derived from living organisms (including cultured cells, bacteria, and viruses), and examples thereof include cells, proteins, nucleic acids, sugars, and lipids.

[0021] The term "biological component adsorption material" refers to a material that can adsorb a target substance in a liquid containing a biological component (hereinafter referred to as a biological fluid) by contacting the liquid with the target substance, thereby reducing the concentration of the target substance in the biological fluid. The target substance to be adsorbed by the biological component adsorption material of the present invention is not particularly limited as long as it is a biological component, but cells or proteins are preferred.

[0022] The shape of the biocomponent adsorption material is not particularly limited, but is preferably a film, particle, hollow fiber, or fiber shape, with hollow fiber or fiber shapes being more preferred. When the biocomponent adsorption material is in fiber shape, an islands-in-the-sea fiber shape is more preferred, as it has a large specific surface area, is flexibly deformable, and is easy to handle. Furthermore, as shapes obtained by processing the above-mentioned fiber shapes, fiber bundles, knitted fabrics, woven fabrics, or nonwoven fabrics are preferred, with knitted fabrics, woven fabrics, or nonwoven fabrics being more preferred, as they have a large specific surface area and low flow path resistance, and knitted fabrics or woven fabrics are even more preferred from the viewpoints of ease of filling into an adsorption column and uniformity of the flow path.

[0023] When the biological component adsorption material is in a fibrous form, from the viewpoint of maintaining mechanical strength, the water-insoluble carrier contained in the biological component adsorption material is preferably a composite fiber, more preferably an islands-in-sea type composite solid fiber, and even more preferably an islands-in-sea type composite solid fiber in which the island component is a water-insoluble polymer and the sea component is a water-insoluble polymer whose main component is different from that of the water-insoluble polymer of the island component.

[0024] Examples of the water-insoluble polymer for the island components of the islands-in-sea type composite solid fiber include polymeric compounds such as polyamide, polyacrylonitrile, polyethylene, polypropylene, nylon, polymethyl methacrylic acid, and polytetrafluoroethylene. These polymeric compounds may be used alone or in combination. Among these, polypropylene is preferred because of its high chemical resistance and excellent thermoplasticity. That is, the islands-in-sea type composite solid fiber is preferably one in which the island components are polypropylene and the sea component is an alloy of polypropylene and a different water-insoluble polymer. When the island components are polypropylene, from the viewpoints of compatibility with polypropylene and ease of ligand bonding, the water-insoluble polymer in the sea component that is different from the water-insoluble polymer for the island components is preferably polystyrene or an alloy of polystyrene and polypropylene (mainly polystyrene).

[0025] "Water-insoluble carrier" refers to a substance that is insoluble in water, including a substrate. Here, "water-insoluble" means that the change in dry weight of the water-insoluble carrier before and after placing it in water is 1% or less. This change in dry weight is the ratio of the dry weight of the solids remaining after immersing the water-insoluble carrier in 37 ° C water in an amount 9 times the dry weight for 1 hour, then removing it with tweezers or the like, and vacuum-drying the remaining water at 50 ° C or less to the dry weight of the water-insoluble carrier before immersion. In the case of a carrier that is not insoluble in water, there is a risk of a large amount of elution when it is brought into contact with a biologically derived fluid, which is undesirable.

[0026] The term "dry weight" refers to the weight of a solid in a dry state. Here, a water-insoluble carrier in a dry state refers to a state in which the amount of liquid components contained in the water-insoluble carrier is 1% by weight or less.

[0027] The term "substrate" refers to a material capable of immobilizing a ligand and that is water-insoluble after the ligand is immobilized. From the viewpoint of stability in biologically derived fluids, the substrate is preferably a polymeric compound. The polymeric compound used as the substrate is not particularly limited; however, since the substrate will be in contact with biologically derived fluids, a sterilization-resistant polymeric compound is preferred. Examples of sterilization-resistant polymeric compounds include vinyl polymers, olefin polymers, sulfone polymers, ester polymers, urethane polymers, imide polymers, amide polymers, polysaccharide polymers, and copolymer polymers. Examples of vinyl polymers include polystyrene, polyvinyl chloride, polymethyl methacrylic acid, polyacrylonitrile, etc. Examples of olefin polymers include polyethylene, polypropylene, polymethylpentene, etc. Examples of sulfone polymers include polysulfone, polyethersulfone, polyarylethersulfone, etc. Examples of ester polymers include polyethylene terephthalate, polybutylene terephthalate, polyarylate, etc. Examples of urethane polymers include polyurethane, etc. Examples of imide polymers include polyimide, polyetherimide, etc. Examples of the amide polymer include nylon 6 and nylon 66. Examples of the polysaccharide polymer include cellulose. Examples of the copolymer include styrene / divinylbenzene copolymer and ethylene / vinyl alcohol copolymer.

[0028] The term "ligand" refers to a chemical structure contained in a water-insoluble carrier in order to exhibit the ability to adsorb a target substance.

[0029] "Ligand having an amino group or a negatively charged functional group" refers to a chemical structure having an amino group or a negatively charged functional group.

[0030] Examples of the ligand having an amino group include a ligand having a polymer containing an amino group such as an ethyleneimine polymer, a ligand having polymyxin B, a ligand having protein A, and a ligand having protein G. The ligand having an amino group is preferably a ligand having an ethyleneimine polymer.

[0031] "Ethylenimine polymer" refers to a polymer of ethyleneimine (CH ２ -CH ２ Examples of the ethyleneimine polymer include ethylenediamine, diethylenetriamine, triethylenetetramine, tetraethylenepentamine, polyethyleneimine (hereinafter referred to as PEI), N-acetylethylenediamine, and N-methyl-2,2'-diaminodiethylamine. The ethyleneimine polymer is preferably diethylenetriamine or PEI.

[0032] "Negatively charged functional group" refers to a functional group having a negative charge. Examples of negatively charged functional groups include sulfonic acid groups, sulfate groups, carboxyl groups, and phosphate groups.

[0033] The term "ligand having a negatively charged functional group" refers to a chemical structure having at least one negatively charged functional group. The negatively charged functional group in the ligand having a negatively charged functional group is not particularly limited, but is preferably a sulfonic acid group or a carboxyl group, and more preferably a thioalkylsulfonic acid group or a thioalkylcarboxyl group.

[0034] The term "thioalkylsulfonic acid group" refers to a chemical structure in which at least one thioether group and one sulfonic acid group are bonded to a chain aliphatic hydrocarbon. Examples of thioalkylsulfonic acid groups include 2-thioethanesulfonic acid and 3-thiopropanesulfonic acid.

[0035] The term "thioalkylcarboxyl group" refers to a chemical structure in which at least one thioether group and one carboxyl group are bonded to a chain aliphatic hydrocarbon. Examples of the thioalkylcarboxyl group include a 2-thiopropionic acid group, a 3-thiopropionic acid group, a 6-thiohexanoic acid group, and a thiosuccinic acid group.

[0036] In view of the above, the ligand having an amino group or a negatively charged functional group is preferably a ligand having an amino group, a sulfonic acid group, or a carboxyl group, and more preferably an ethyleneimine polymer, a ligand having a thioalkylsulfonic acid group, or a thioalkylcarboxyl group.

[0037] The ligand having an amino group or a negatively charged functional group can have an optimal chemical structure depending on the target substance. For example, when the target substance is endotoxin, a ligand having polymyxin B can be used as the ligand having an amino group or a negatively charged functional group. When the target substance is a cytokine such as high-mobility group box 1 (hereinafter referred to as HMGB1) or oxidized low-density lipoprotein, a ligand having PEI can be used. Furthermore, when the target substance is soluble fms-like tyrosine kinase 1 (hereinafter referred to as sFlt-1), a ligand having a thioalkylsulfonic acid group or a thioalkylcarboxyl group can be used. When the target substance is latent TGF-β, a ligand having diethylenetriamine can be used. When the target substance is an antibody, a ligand having protein A or protein G can be used. Furthermore, depending on the target substance, ligands having peptides, proteins, antibodies, nucleic acid aptamers modified with amino groups or negatively charged functional groups, etc. can also be used.

[0038] The ligand having an amino group or a negatively charged functional group preferably contains an amidomethyl group, more preferably an acetamidomethyl group, for the purpose of binding to a substrate.

[0039] "Chemical structure containing a halogen atom" means a chemical structure containing one or more halogen atoms as a partial structure. Examples of chemical structures containing a halogen atom include a halomethyl group, a haloacetyl group, a haloacetamidomethyl group, and a halogenated alkyl group. The halogen atom in the chemical structure containing a halogen atom is not particularly limited, but a chlorine atom is preferred. Furthermore, a chloroacetamidomethyl group is preferred as a chemical structure containing a halogen atom.

[0040] The ligand having an amino group or a negatively charged functional group and the chemical structure containing a halogen atom may each be bound to a substrate, or a chemical structure containing both the ligand having an amino group or a negatively charged functional group and the chemical structure containing a halogen atom may be bound to a substrate.

[0041] The binding mode of the ligand having an amino group or a negatively charged functional group and the chemical structure containing a halogen atom to the substrate is not particularly limited, and examples thereof include a covalent bond, an electrostatic interaction, a hydrophobic interaction, a hydrogen bond, etc. From the viewpoint of reducing the risk of elution into a biologically derived fluid, the binding mode to the substrate is preferably a covalent bond or an electrostatic interaction.

[0042] From the viewpoint of exhibiting target substance adsorption performance, the content of amino groups or negatively charged functional groups per 1 g of dry weight of the water-insoluble carrier is preferably 0.04 mmol or more. The upper limit of the content of amino groups or negatively charged functional groups is not particularly limited, but is preferably 0.50 mmol, more preferably 0.24 mmol, per 1 g of dry weight of the water-insoluble carrier. That is, the content of amino groups or negatively charged functional groups per 1 g of dry weight of the water-insoluble carrier is preferably 0.04 to 0.50 mmol, more preferably 0.04 to 0.24 mmol.

[0043] The content of halogen atoms per 1 g of dry weight of the water-insoluble carrier is preferably 0.05 mmol or more, more preferably 0.06 mmol or more. Furthermore, from the viewpoint of suppressing nonspecific adsorption of proteins, the content of halogen atoms per 1 g of dry weight of the water-insoluble carrier is preferably 2.00 mmol or less, more preferably 1.50 mmol or less, and even more preferably 1.38 mmol or less. That is, the content of halogen atoms per 1 g of dry weight of the water-insoluble carrier is preferably 0.05 to 2.00 mmol, more preferably 0.06 to 1.50 mmol, and even more preferably 0.06 to 1.38 mmol.

[0044] The biological component adsorption material of the present invention can be produced, for example, by the following method. A substrate is immersed in a reaction solution in which a compound that forms the basis of a chemical structure containing a halogen atom is dissolved, and then washed to produce a substrate (hereinafter referred to as an intermediate) on which a chemical structure containing a halogen atom is immobilized. The intermediate is then immersed in a reaction solution in which a compound that forms the basis of a ligand having an amino group or a negatively charged functional group is dissolved, and then washed to produce the biological component adsorption material.

[0045] The biological component adsorption material can also be produced by immersing a substrate in a reaction solution in which a compound that forms the basis of a chemical structure containing a halogen atom and a compound that forms the basis of a ligand having an amino group or a negatively charged functional group are dissolved, or in a reaction solution in which a compound that forms the basis of a chemical structure containing a halogen atom and a ligand having an amino group or a negatively charged functional group is dissolved, and then washing the substrate.

[0046] When the substrate is formed into a fiber, film, hollow fiber, particle, or other shape, a commercially available substrate can be purchased, heated and melted, and then extruded from a nozzle, or dissolved in a good solvent and then extruded from a nozzle into a poor solvent or the atmosphere, thereby forming the substrate into any desired shape.

[0047] In the reaction between a compound that serves as the basis for a chemical structure containing a halogen atom and a substrate (hereinafter referred to as a halogen atom introduction reaction), for example, N-methylol-α-chloroacetamide (hereinafter referred to as NMCA) can be used as the compound that serves as the basis for a chemical structure containing a halogen atom.

[0048] As the solvent for the reaction solution for the halogen atom introduction reaction, for example, a mixed solvent of nitrobenzene and sulfuric acid can be used. When the polymer compound used as the substrate is polystyrene, an alloy of polystyrene and polypropylene (mainly polystyrene), or a copolymer containing polystyrene, nitrobenzene penetrates into the substrate and swells it, so by reducing the ratio of nitrobenzene in the mixed solvent, the amount of introduced chemical structure containing halogen atoms can be kept within an appropriate range. For the above reasons, the mass ratio of nitrobenzene to sulfuric acid (nitrobenzene:sulfuric acid) in the mixed solvent is preferably 37.5:62.5.

[0049] The reaction temperature for the halogen atom introduction reaction is preferably 0 to 50°C, more preferably 0 to 20°C.

[0050] In the halogen atom introduction reaction, paraformaldehyde (hereinafter referred to as PFA) or a solution in which PFA is dissolved may be added to the reaction solution. When a solution in which PFA is dissolved is added, the solvent in which PFA is dissolved is not particularly limited, but is preferably the same as the solvent in the reaction solution.

[0051] As the washing solvent used in the first washing operation immediately after the halogen atom introduction reaction, for example, nitrobenzene or methanol can be used. When the polymer compound used as the substrate is polystyrene, an alloy of polystyrene and polypropylene (mainly polystyrene), or a copolymer containing polystyrene, among these, methanol can be used to shrink the substrate, and therefore the amount of introduced chemical structure containing halogen atoms can be reduced compared to when nitrobenzene is used.

[0052] In the reaction between the intermediate and a compound that will become a ligand having an amino group or a negatively charged functional group (hereinafter referred to as the ligand introduction reaction), for example, an ethyleneimine polymer, a thioalkylsulfonic acid, or a thioalkylcarboxylic acid can be used as the compound that will become a ligand having an amino group or a negatively charged functional group.

[0053] As a reaction solvent for the ligand introduction reaction, for example, water or dimethyl sulfoxide can be used.

[0054] The reaction temperature for the ligand introduction reaction is preferably 10 to 80°C.

[0055] The amino group content per 1 g of dry weight of the water-insoluble carrier contained in the biological component adsorption material can be determined, for example, by immersing the biological component adsorption material in an aqueous solution, extracting only the water-insoluble carrier, measuring the dry weight, and then ion-exchanging the amino groups in the water-insoluble carrier with hydrochloric acid and back-titrating with a sodium hydroxide solution.

[0056] The content of negatively charged functional groups per gram of dry weight of the water-insoluble carrier contained in the biological component adsorption material can be determined, for example, by immersing the biological component adsorption material in an aqueous solution, extracting only the water-insoluble carrier, measuring the dry weight, and then ion-exchanging the negatively charged functional groups in the water-insoluble carrier with an aqueous sodium hydroxide solution and back-titrating with hydrochloric acid.

[0057] The content of halogen atoms per gram of dry weight of the water-insoluble carrier contained in the biological component adsorption material can be determined, for example, by immersing the biological component adsorption material in an aqueous solution, extracting only the water-insoluble carrier, and measuring it by X-ray fluorescence analysis or X-ray photoelectron spectroscopy. Furthermore, when the chemical structure containing halogen atoms is a haloacetamidomethyl group, the dry weight of the water-insoluble carrier is measured, and then the water-insoluble carrier is heated in hydrochloric acid to hydrolyze it, and the resulting amino groups are ion-exchanged with hydrochloric acid and back-titrated with sodium hydroxide solution to quantify the content of amide groups in the water-insoluble carrier, and the content of the amide groups can be regarded as the content of halogen atoms. Furthermore, when the ligand having an amino group is an acetamidomethyl group having an amino group, and the chemical structure containing halogen atoms is a haloacetamidomethyl group, the content of halogen atoms can also be determined by dividing the content of amino groups separately determined by the above-mentioned procedure by the valence of the amino group of the ligand having an amino group and subtracting this value from the content of amide groups. In addition, when the ligand having a negatively charged functional group is an acetamidomethyl group having a negatively charged functional group and the chemical structure containing a halogen atom is a haloacetamidomethyl group, the content of the halogen atom can also be determined by dividing the content of the negatively charged functional group determined separately by the above-mentioned operation by the valence of the negatively charged functional group of the ligand having a negatively charged functional group and subtracting the result from the content of the amide group.

[0058] A method for measuring the adsorption performance of a biological component adsorption material for a target substance includes, for example, impregnating the biological component adsorption material with a bovine serum albumin (hereinafter, BSA) solution or a fetal bovine serum (hereinafter, FBS) solution in which the target substance has been dissolved or dispersed, measuring the concentration of the target substance in the BSA solution or FBS solution before and after impregnation, and calculating the adsorption rate of the target substance from the concentration of the target substance before and after impregnation.

[0059] The target substance to be adsorbed is not particularly limited, but when used for blood purification, it is preferably a pathogenic substance of any disease and a substance present in blood. When the disease is sepsis or acute respiratory distress syndrome, the target substance is preferably a cytokine, and among cytokines, interleukin-6, interleukin-8, or HMGB1 is preferred. When the disease is preeclampsia, the target substance is preferably sFlt-1, soluble endoglin, or HMGB1. When the disease is cancer, the target substance is preferably latent TGF-β or LAP-positive immune cells.

[0060] A method for measuring the nonspecific adsorption performance of a biological component adsorption material for proteins includes, for example, impregnating the biological component adsorption material with a phosphate buffer solution containing dissolved proteins that are abundant in blood, plasma, and serum, measuring the protein concentrations in the phosphate buffer solution before and after impregnation, and calculating the protein adsorption rate from the protein concentrations before and after impregnation.

[0061] The protein used in measuring the nonspecific adsorption performance of a protein is not particularly limited, but is preferably albumin, which is present in the largest amount in blood, plasma, and serum.

[0062] The present invention will be described below with reference to examples, but the present invention is not limited to these examples.

[0063] In the following, NMCA was used as the compound that serves as the basis for the chemical structure containing a halogen atom, PEI or diethylenetriamine was used as the compound that serves as the basis for the ligand having an amino group, and sodium 3-mercaptopropanesulfonate, 3-mercaptopropionic acid, or mercaptosuccinic acid was used as the compound that serves as the basis for the ligand having a negatively charged functional group.

[0064] (Production of knitted fabric A) Using a spinneret having discharge holes designed to form a fiber cross section in which 264 island components exist in the sea component, melt spinning was performed using a mixture of 90 mass % polystyrene and 10 mass % polypropylene for the sea component and polypropylene for the island components, with the discharge ratio of the sea component to the island components being 50:50 by weight, to produce knitted fabric A using fibers with a total fineness of 108 dtex, each of which was a bundle of 36 islands-in-sea type composite fibers, each of which had a fineness of 3 dtex.

[0065] (Preparation of intermediate A) A chemical structure containing a halogen atom was introduced into knitted fabric A by the following procedure. 0.39 g of PFA was added to a mixed solution of 90 g of nitrobenzene and 150 g of sulfuric acid, and the mixture was dissolved in an ice bath. 18.1 g of NMCA was then added, and the mixture was stirred and dissolved in an ice bath for 2 hours. 10 g of knitted fabric A was immersed in this solution and allowed to react in the ice bath for 2 hours. After the reaction, knitted fabric A was removed, immersed in methanol, which was the first washing solution, and washed. The mixture was then substituted with methanol seven times, washed, and washed with ion-exchanged water to obtain intermediate A.

[0066] (Preparation of Intermediate B) Intermediate B was obtained by the same procedure as in the preparation of Intermediate A, except that the amount of PFA added was changed from 0.39 g to 0.52 g.

[0067] (Preparation of Intermediate C) Intermediate C was obtained by the same procedure as in the preparation of Intermediate A, except that the first washing solution was changed from methanol to nitrobenzene.

[0068] (Preparation of Intermediate D) Intermediate D was obtained by the same procedure as in the preparation of Intermediate A, except that the amount of PFA added was changed from 0.39 g to 0.52 g and the first washing solution was changed from methanol to nitrobenzene.

[0069] (Preparation of intermediate E) 0.26 g of PFA was added to a mixed solution of 120 g of nitrobenzene and 120 g of sulfuric acid, and dissolved in an ice bath. 18.1 g of NMCA was then added, and the mixture was stirred in an ice bath for 2 hours to dissolve. 10 g of knitted fabric A was immersed in this solution and allowed to react in an ice bath for 2 hours. After the reaction, knitted fabric A was removed, immersed in methanol, which was the first washing solution, and washed. The mixture was then substituted with methanol seven times, washed, and washed with ion-exchanged water to obtain intermediate E.

[0070] (Preparation of Intermediate F) Intermediate F was obtained by the same procedure as in the preparation of Intermediate E, except that the first washing solution was changed from methanol to nitrobenzene.

[0071] (Preparation of Biological Component Adsorption Material A) A ligand containing an amino group was introduced into intermediate A by the following procedure. 1.5 g of PEI with a molecular weight of approximately 10,000, serving as the compound that will form the basis of the amino group-containing ligand, was added to and dissolved in 350 mL of distilled water to prepare a 0.43 wt % PEI aqueous solution. 10 g of intermediate A was immersed in this PEI aqueous solution and stirred for 3 hours. Intermediate A was then immersed in ion-exchanged water and thoroughly washed to obtain biological component adsorption material A. The series of operations from preparing the PEI aqueous solution to washing the knitted fabric with ion-exchanged water were carried out at a solution temperature of 40°C.

[0072] (Preparation of Biological Component Adsorbing Material B) Biological component adsorbing material B was obtained in the same manner as in the preparation of biological component adsorbing material A, except that the amount of PEI added was changed from 1.5 g to 7.5 g.

[0073] (Preparation of Biological Component Adsorbing Material C) Biological component adsorbing material C was obtained in the same manner as in the preparation of biological component adsorbing material A, except that the amount of PEI added was changed from 1.5 g to 37.6 g.

[0074] (Preparation of Biocomponent Adsorbent Material D) The same procedure as for preparing Biocomponent Adsorbent Material A was carried out, except that intermediate A was changed to intermediate B and the amount of PEI added was changed from 1.5 g to 37.6 g, to obtain Biocomponent Adsorbent Material D.

[0075] (Preparation of Biological Component Adsorbing Material E) Biological component adsorbing material E was obtained in the same manner as in the preparation of biological component adsorbing material A, except that intermediate A was replaced with intermediate C.

[0076] (Preparation of Biocomponent Adsorption Material F) The same procedure as for preparing Biocomponent Adsorption Material A was carried out, except that intermediate A was changed to intermediate C and the amount of PEI added was changed from 1.5 g to 7.5 g, to obtain Biocomponent Adsorption Material F.

[0077] (Preparation of Biocomponent Adsorbent Material G) The same procedure as for preparing Biocomponent Adsorbent Material A was carried out, except that intermediate A was changed to intermediate C and the amount of PEI added was changed from 1.5 g to 37.6 g, to obtain Biocomponent Adsorbent Material G.

[0078] (Preparation of Biocomponent Adsorption Material H) The same procedure as for preparing Biocomponent Adsorption Material A was carried out, except that intermediate A was changed to intermediate D and the amount of PEI added was changed from 1.5 g to 37.6 g, to obtain Biocomponent Adsorption Material H.

[0079] (Preparation of Biocomponent Adsorbing Material I) The same procedure as for preparing Biocomponent Adsorbing Material A was carried out, except that Intermediate A was changed to Intermediate E and the amount of PEI added was changed from 1.5 g to 37.6 g, to obtain Biocomponent Adsorbing Material I.

[0080] (Preparation of Biocomponent Adsorbent Material J) The same procedure as for preparing Biocomponent Adsorbent Material A was carried out, except that intermediate A was changed to intermediate F and the amount of PEI added was changed from 1.5 g to 37.6 g, to obtain Biocomponent Adsorbent Material J.

[0081] (Preparation of Biocomponent Adsorbent Material K) A ligand containing a negatively charged functional group was introduced into Intermediate A by the following procedure. 5.0 g of sodium 3-mercaptopropanesulfonate, which serves as the compound underlying the ligand having a negatively charged functional group, and 14 mL of 0.1 N aqueous sodium hydroxide solution were added and dissolved in 267 mL of distilled water to prepare a 100 mM aqueous solution of sodium 3-mercaptopropanesulfonate. 1 g of Intermediate A was immersed in this aqueous solution of sodium 3-mercaptopropanesulfonate and stirred for 3 hours. Intermediate A was then immersed in ion-exchanged water and thoroughly washed to obtain Biocomponent Adsorbent Material K. The series of operations from preparing the aqueous solution of sodium 3-mercaptopropanesulfonate to washing the knitted fabric with ion-exchanged water were carried out at a solution temperature of 60°C.

[0082] (Preparation of Biological Component Adsorbing Material L) Biological component adsorbing material L was obtained in the same manner as in the preparation of biological component adsorbing material K, except that intermediate A was replaced with intermediate B.

[0083] (Preparation of Biological Component Adsorbing Material M) Biological component adsorbing material M was obtained in the same manner as in the preparation of biological component adsorbing material K, except that intermediate A was replaced with intermediate C.

[0084] (Preparation of Biological Component Adsorbing Material N) Biological component adsorbing material N was obtained in the same manner as in the preparation of biological component adsorbing material K, except that intermediate A was replaced with intermediate D.

[0085] (Preparation of Biological Component Adsorbing Material O) Biological component adsorbing material O was obtained in the same manner as in the preparation of biological component adsorbing material K, except that intermediate A was replaced with intermediate E.

[0086] (Preparation of Biological Component Adsorption Material P) A ligand containing a negatively charged functional group was introduced into Intermediate A by the following procedure. 3-mercaptopropionic acid, as a compound that serves as the basis for the ligand having a negatively charged functional group, and an aqueous sodium hydroxide solution were added to distilled water to final concentrations of 100 mM and 200 mM, respectively, to prepare a reaction solution. 1 g of Intermediate A was immersed in 281 mL of this reaction solution and stirred for 3 hours. Intermediate A was then immersed in ion-exchanged water and thoroughly washed to obtain Biological Component Adsorption Material P. A series of operations from preparing the reaction solution to washing the knitted fabric with ion-exchanged water were carried out at a solution temperature of 60°C.

[0087] (Preparation of Biological Component Adsorbing Material Q) A biological component adsorbing material Q was obtained in the same manner as in the preparation of the biological component adsorbing material P, except that intermediate A was replaced with intermediate B.

[0088] (Preparation of Biocomponent Adsorption Material R) Biocomponent adsorption material R was obtained by carrying out the same operations as in the preparation of biocomponent adsorption material P, except that the compound that serves as the basis for the ligand having a negatively charged functional group was changed to mercaptosuccinic acid, the final concentration of the compound that serves as the basis for the ligand having a negatively charged functional group in the reaction solution was changed to 1000 mM, and the final concentration of sodium hydroxide was changed to 2200 mM.

[0089] (Preparation of Biological Component Adsorbing Material S) The same procedure as in the preparation of the biological component adsorbing material R was carried out, except that intermediate A was changed to intermediate B, to obtain biological component adsorbing material S.

[0090] (Preparation of Biological Component Adsorbing Material T) A biological component adsorbing material T was obtained in the same manner as in the preparation of the biological component adsorbing material P, except that intermediate A was replaced with intermediate E.

[0091] (Preparation of Biological Component Adsorbing Material U) Biological component adsorbing material U was obtained in the same manner as in the preparation of biological component adsorbing material R, except that intermediate A was replaced with intermediate E.

[0092] (Preparation of Biological Component Adsorption Material V) A ligand containing an amino group was introduced into Intermediate A by the following procedure. Diethylenetriamine, the compound that forms the basis of the amino group-containing ligand, was added to distilled water to a final concentration of 1500 mM to prepare a reaction solution. 10 g of Intermediate A was immersed in 350 mL of this reaction solution and stirred for 3 hours. Intermediate A was then immersed in ion-exchanged water and thoroughly washed to obtain Biological Component Adsorption Material V. A series of operations from preparing the reaction solution to washing the knitted fabric with ion-exchanged water were carried out at a solution temperature of 40°C.

[0093] (Preparation of Biological Component Adsorbing Material W) Biological component adsorbing material W was obtained in the same manner as in the preparation of biological component adsorbing material V, except that intermediate A was replaced with intermediate B.

[0094] (Preparation of Biological Component Adsorbing Material X) Biological component adsorbing material X was obtained in the same manner as in the preparation of biological component adsorbing material V, except that intermediate A was changed to intermediate E.

[0095] (Measurement of Amino Group Content of Water-Insoluble Carrier) The amino group content of the water-insoluble carrier was determined by measuring the amount of amino groups in the water-insoluble carrier by acid-base back titration. The detailed method is described below.

[0096] To a polypropylene container, 1.0 g of the biocomponent adsorption material and 50 mL of 6 M sodium hydroxide aqueous solution were added and stirred for 30 minutes, and the water-insoluble carrier was filtered off using filter paper. The filtered water-insoluble carrier was then added to 50 mL of ion-exchanged water, stirred for 30 minutes, and filtered off using filter paper. The desalted water-insoluble carrier was obtained by repeatedly adding the water-insoluble carrier to ion-exchanged water and filtering it until the pH of the ion-exchanged water containing the water-insoluble carrier reached 7. The desalted water-insoluble carrier was then allowed to stand under reduced pressure at 50°C for 12 hours or more, after which 1.0 g of the water-insoluble carrier and 30 mL of 0.1 M hydrochloric acid were added to a polypropylene container and stirred for 10 minutes. After stirring, 5 mL of the solution alone was removed and transferred to a polypropylene container. 0.1 mL of 0.1 M sodium hydroxide aqueous solution was added dropwise to the solution. After addition, the solution was stirred for 10 minutes, and the pH of the solution was measured. The amount of sodium hydroxide solution added when the pH of the solution exceeded 8.5 was defined as the titer per gram of the aqueous sodium hydroxide solution. The amino group content per gram of the dry weight of the water-insoluble carrier was calculated using the following formula 1.

[0097] Amino group content (mmol / g) per 1 g of dry weight of water-insoluble carrier = [{amount of 0.1 M hydrochloric acid added (30 mL) / amount of hydrochloric acid removed (5 mL)} × {amount of neutralization solution (5 mL) - amount of titration (mL)} × concentration of aqueous sodium hydroxide solution (0.1 M)] ÷ dry weight of added water-insoluble carrier (1.0 g) ... Equation 1

[0098] (Measurement of the content of negatively charged functional groups in water-insoluble carriers) The content of negatively charged functional groups in water-insoluble carriers was determined by measuring the negatively charged functional groups in the water-insoluble carriers by acid-base back titration. The detailed method is described below.

[0099] To a polypropylene container, 1.0 g of the biocomponent adsorption material and 50 mL of 6 M hydrochloric acid were added and stirred for 30 minutes, and the water-insoluble carrier was filtered off using filter paper. The filtered water-insoluble carrier was then added to 50 mL of ion-exchanged water, stirred for 30 minutes, and filtered off using filter paper. The desalted water-insoluble carrier was obtained by repeatedly adding the water-insoluble carrier to ion-exchanged water and filtering it until the pH of the ion-exchanged water containing the water-insoluble carrier reached 7. The desalted water-insoluble carrier was then allowed to stand under reduced pressure at 50°C for 12 hours or more, after which 1.0 g of the water-insoluble carrier and 30 mL of 0.1 M aqueous sodium hydroxide solution were added to a polypropylene container and stirred for 10 minutes. After stirring, 5 mL of the solution was removed and transferred to a polypropylene container. 0.1 mL of 0.1 M hydrochloric acid was added dropwise to the solution. After addition, the solution was stirred for 10 minutes, and the pH of the solution was measured. The amount of hydrochloric acid added when the pH of the solution reached 5.5 or less was taken as the titer per gram. The content of negatively charged functional groups per gram of dry weight of the water-insoluble carrier was calculated using the following formula 2.

[0100] Content of negatively charged functional groups per 1 g of dry weight of water-insoluble carrier (mmol / g) = [{amount of added 0.1 M aqueous sodium hydroxide solution (30 mL) / amount of removed aqueous sodium hydroxide solution (5 mL)} × {amount of neutralization solution (5 mL) - amount of titration (mL)} × concentration of hydrochloric acid (0.1 M)] ÷ dry weight of added water-insoluble carrier (1.0 g) ... Equation 2

[0101] (Measurement of chlorine atom content of water-insoluble carrier) The content of chlorine atoms contained in the water-insoluble carrier was calculated as follows. First, the content of amino groups derived from amide groups generated by hydrolysis of chloroacetamidomethyl groups and acetamidomethyl groups having amino groups or negatively charged functional groups in the water-insoluble carrier was measured by acid-base back titration, and the content of amide groups in the water-insoluble carrier was calculated. The separately measured content of amino groups or negatively charged functional groups was divided by the valence of the amino groups or negatively charged functional groups of the ligand having an amino group or negatively charged functional group, and the value was subtracted from the obtained content of amide groups to determine the content of chlorine atoms. The detailed method is described below.

[0102] 1.0 g of the biocomponent adsorption material and 100 mL of 6 M hydrochloric acid were added to a 200 mL recovery flask and refluxed at 130 ° C. for 24 hours. After refluxing, the water-insoluble carrier was recovered by filtering with filter paper to obtain a water-insoluble carrier after acid hydrolysis. Next, the entire amount of the obtained water-insoluble carrier after acid hydrolysis and 50 mL of 6 M sodium hydroxide aqueous solution were added to a polypropylene container, stirred for 30 minutes, and then filtered using filter paper. The filtered water-insoluble carrier after acid hydrolysis was then added to 50 mL of ion-exchanged water, stirred for 30 minutes, and filtered using filter paper. Addition to ion-exchanged water and filtration were repeated until the pH of the ion-exchanged water containing the water-insoluble carrier after acid hydrolysis reached 7. After the pH of the ion-exchanged water containing the water-insoluble carrier after acid hydrolysis reached 7, the water-insoluble carrier after acid hydrolysis was left to stand at 50 ° C. under reduced pressure for 12 hours or more and dried. Next, the entire amount of the water-insoluble carrier and 60 mL of 0.1 M hydrochloric acid were added to a polypropylene container and stirred for 10 minutes. After stirring, 5 mL of the solution alone was extracted and transferred to a polypropylene container. 0.1 mL of 0.1 M aqueous sodium hydroxide solution was added dropwise to the solution. After addition, the solution was stirred for 10 minutes and the pH of the solution was measured. The dropwise addition of the aqueous sodium hydroxide solution and the stirring after addition were repeated, and the amount of the aqueous sodium hydroxide solution added when the pH of the solution exceeded 8.5 was taken as the titer per 1.0 g. The content of amide groups per 1.0 g of dry weight of the water-insoluble carrier was calculated using the following formula 3.

[0103] Amide group content (mmol / g) per 1 g of dry weight of water-insoluble carrier = [{amount of 0.1 M hydrochloric acid added (60 mL) / amount of hydrochloric acid removed (5 mL)} × {amount of neutralization solution (5 mL) - amount of titration (mL)} × concentration of aqueous sodium hydroxide solution (0.1 M)] ÷ dry weight of added water-insoluble carrier (1.0 g) ... Equation 3

[0104] In the case of a water-insoluble carrier containing a ligand having an amino group, the content of chlorine atoms contained in the water-insoluble carrier was calculated using the following formula 4.

[0105] Chlorine atom content (mmol / g) per 1 g of dry weight of water-insoluble carrier containing a ligand having an amino group=amide group content (mmol / g) per 1 g of dry weight of water-insoluble carrier−{amino group content (mmol / g) per 1 g of dry weight of water-insoluble carrier / valence of ligand having an amino group} Equation 4

[0106] In the case of a water-insoluble carrier containing a ligand having a negatively charged functional group, the content of chlorine atoms contained in the water-insoluble carrier was calculated using the following formula 5.

[0107] Chlorine atom content (mmol / g) per 1 g of dry weight of a water-insoluble carrier containing a ligand having a negatively charged functional group = amide group content (mmol / g) per 1 g of dry weight of a water-insoluble carrier - {negatively charged functional group content (mmol / g) per 1 g of dry weight of a water-insoluble carrier ÷ valence of a ligand having a negatively charged functional group} Equation 5

[0108] (Measurement of Albumin Adsorption Rate) In order to confirm the albumin adsorption capacity of the biological component adsorption material, the albumin adsorption rate was measured by the following method.

[0109] The biological component adsorption material was cut into disks with a diameter of 10 mm, and four disks were placed in a polypropylene container and washed with physiological saline. After washing, the biological component adsorption material was dehydrated and transferred to a new polypropylene container (hereinafter referred to as container A containing the biological component adsorption material).

[0110] Next, human serum albumin (hereinafter referred to as albumin) was added to phosphate buffered saline to prepare an albumin solution at 100 μg / mL. 1 mL of the 100 μg / mL albumin solution was added to container A containing the biological component adsorption material, and the container was shaken using a rotator at 37°C and 6 rpm for 1 hour.

[0111] After shaking, the albumin solution remaining in container A containing the biological component adsorption material (hereinafter referred to as post-shaking solution A) was placed into a new polypropylene container. Albumin in the post-shaking solution was detected using a BCA assay kit, and the albumin concentration was quantified by measuring the absorbance at 562 nm using a microplate reader SpectraMax M5 (Molecular Devices Japan).

[0112] One mL of a 100 μg / mL albumin solution was added to a polypropylene container containing no biological component adsorption material, and the container was shaken in the same manner as for container A containing the biological component adsorption material to obtain blank solution A. After shaking, the albumin concentration of blank solution A was quantified in the same manner as for solution A, and the albumin adsorption rate of the biological component adsorption material was calculated using equation 6.

[0113] Albumin adsorption rate (%) = {(albumin concentration of blank solution A) - (albumin concentration of solution A after shaking)} / (albumin concentration of blank solution A) × 100 Equation 6

[0114] (Measurement of HMGB1 Adsorption Rate) In order to confirm the HMGB1 adsorption capacity of the biological component adsorption material, the HMGB1 adsorption rate was measured by the following method.

[0115] The biological component adsorption material was cut into a disk with a diameter of 10 mm, and one disk was placed in a polypropylene container and washed with physiological saline. After washing, the biological component adsorption material was dehydrated and transferred to a new polypropylene container (hereinafter referred to as container B containing the biological component adsorption material).

[0116] Next, 0.53 g of 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 the 3.5% BSA solution to obtain a 100 ng / mL HMGB1 solution. 414 μL of the 100 ng / mL HMGB1 solution was added to container B containing the biocomponent adsorption material, and the mixture was shaken at 37°C and 500 rpm for 2 hours using a thermomixer (Eppendorf).

[0117] After shaking, the HMGB1 solution remaining in container B containing the biological component adsorption material (hereinafter referred to as post-shaking solution B) was placed into a new polypropylene container. HMGB1 in post-shaking solution B 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.).

[0118] 414 μL of a 100 ng / mL HMGB1 solution was added to a polypropylene container containing no biological component adsorption material, and the container was shaken in the same manner as for container B containing the biological component adsorption material to obtain blank solution B. After shaking, the HMGB1 concentration of blank solution B was quantified in the same manner as for solution B, and the HMGB1 adsorption rate of the biological component adsorption material was calculated using equation 7.

[0119] HMGB1 adsorption rate (%)={(HMGB1 concentration in blank solution B)−(HMGB1 concentration in solution B after shaking)} / (HMGB1 concentration in blank solution B)×100 Equation 7

[0120] (Measurement of sFlt-1 Adsorption Rate) In order to confirm the sFlt-1 adsorption capacity of the biological component adsorption material, the sFlt-1 adsorption rate was measured by the following method.

[0121] The biological component adsorption material was cut into disks with a diameter of 10 mm, and three disks were placed in a polypropylene container and washed with physiological saline. After washing, the biological component adsorption material was dehydrated and transferred to a new polypropylene container (hereinafter referred to as container C containing the biological component adsorption material).

[0122] Next, 100 μg of Human VEGFR1 / FLT-1 Protein (His Tag) (Shino Biological Corporation) was dissolved in 400 μL of distilled water and diluted with FBS to obtain an 8 ng / mL sFlt-1 solution. 745 μL of the 8 ng / mL sFlt-1 solution was added to container C containing the biological component adsorption material, and the container was shaken at 37°C and 20 rpm using a rotator for 2 hours.

[0123] After shaking, the sFlt-1 solution remaining in container C containing the biocomponent adsorption material (hereinafter referred to as post-shaking solution C) was placed into a new polypropylene container. sFlt-1 in post-shaking solution C was detected using a Human VEGFR1 / Flt-1 DuoSet (manufactured by R&D Co.), and the sFlt-1 concentration was quantified by measuring the absorbance at 450 nm and 540 nm using a microplate reader SpectraMax M5 (manufactured by Molecular Devices Japan).

[0124] 745 μL of an 8 ng / mL sFlt-1 solution was added to a polypropylene container containing no biological component adsorption material, and the container was shaken in the same manner as for container C containing the biological component adsorption material to obtain blank solution C. After shaking, the sFlt-1 concentration of blank solution C was quantified in the same manner as for solution C, and the sFlt-1 adsorption rate of the biological component adsorption material was calculated using equation 8.

[0125] sFlt-1 adsorption rate (%)={(sFlt-1 concentration in blank solution C)−(sFlt-1 concentration in solution C after shaking)} / (sFlt-1 concentration in blank solution C)×100 (Equation 8)

[0126] (Measurement of Latent TGF-β Adsorption Rate) In order to confirm the Latent TGF-β adsorption capacity of the biological component-adsorbing material, the Latent TGF-β adsorption rate was measured by the following method.

[0127] The biological component adsorption material was cut into disks with a diameter of 10 mm, and five disks were placed in a polypropylene container and washed with physiological saline. After washing, the biological component adsorption material was dehydrated and transferred to a new polypropylene container (hereinafter referred to as container D containing the biological component adsorption material).

[0128] Next, Latent TGF-β1 (Recombinant Human Latent TGF-beta; manufactured by R&D) was dissolved in 3.5% BSA to obtain a 25 ng / mL Latent TGF-β solution. 1,100 μL of the 25 ng / mL Latent TGF-β solution was added to container D containing the biocomponent adsorption material, and the container was shaken at 37°C and 20 rpm using a rotator for 2 hours.

[0129] After shaking, the Latent TGF-β solution remaining in Container D containing the biocomponent adsorption material (hereinafter referred to as "Solution D after shaking") was placed in a new polypropylene container. Latent TGF-β in Solution D after shaking was detected using a Quantikine Human LAP (TGF-β1) ELISA Kit (manufactured by R&D), and the concentration of Latent TGF-β was quantified by measuring the absorbance at 450 nm and 540 nm using a microplate reader SpectraMax M5 (manufactured by Molecular Devices Japan).

[0130] 1100 μL of a 25 ng / mL Latent TGF-β solution was added to a polypropylene container containing no biological component adsorbing material, and the container was shaken in the same manner as for container D containing the biological component adsorbing material to obtain blank solution D. After shaking, the Latent TGF-β concentration of blank solution D was quantified in the same manner as for solution D, and the Latent TGF-β adsorption rate of the biological component adsorbing material was calculated using equation 9.

[0131] Latent TGF-β adsorption rate (%)={(Latent TGF-β concentration in blank solution D)−(Latent TGF-β concentration in solution D after shaking)} / (Latent TGF-β concentration in blank solution D)×100 (Equation 9)

[0132] Examples 1 to 8 and Comparative Examples 1 to 3 The chlorine atom content, amino group content, albumin adsorption rate, and HMGB1 adsorption rate were measured for the biological component adsorption materials A to J and intermediate E. The results are shown in Table 1.

[0133]

[0134] The results of Examples 1 to 8 and Comparative Examples 1 to 3 in Table 1 reveal that materials for adsorbing biological components having a halogen content per gram of dry weight of the water-insoluble carrier below a certain level have a low albumin adsorption rate and suppress nonspecific adsorption of proteins.

[0135] Furthermore, it was revealed that if the content of amino groups per 1 g of dry weight of the water-insoluble carrier is 0.04 mmol or more, the material for adsorbing biological components has a high adsorption rate of HMGB1 and can exhibit adsorption performance for target substances.

[0136] The results of Examples 1 to 4 and Comparative Example 1 and the results of Examples 5 to 8 and Comparative Example 2 in Table 1 reveal that by using methanol as the first washing solvent when preparing an intermediate, the content of halogen atoms can be reduced compared to when the washing solvent is nitrobenzene.

[0137] Examples 9 to 16 and Comparative Examples 4 to 9 The chlorine atom content, negatively charged functional group content, albumin adsorption rate, and sFlt-1 adsorption rate were measured for biological component adsorption materials K to U and intermediates A, B, and E. The results are shown in Table 2.

[0138]

[0139] The results of Examples 9 to 16 and Comparative Examples 4 to 9 in Table 2 reveal that materials for adsorbing biological components having a halogen content per gram of dry weight of the water-insoluble carrier below a certain level have a low albumin adsorption rate and suppress nonspecific adsorption of proteins.

[0140] Furthermore, it was revealed that if the content of negatively charged functional groups per gram of dry weight of the water-insoluble carrier is 0.09 mmol or more, the material for adsorbing biological components has a high adsorption rate of sFlt-1 and can exhibit adsorption performance for target substances.

[0141] Examples 17 and 18 and Comparative Examples 10 and 11 The chlorine atom content, amino group content, albumin adsorption rate, and latent TGF-β adsorption rate were measured for biological component-adsorbing materials V, W, and X, and intermediate E. The results are shown in Table 3.

[0142]

[0143] The results of Examples 17 and 18 and Comparative Examples 10 and 11 in Table 3 reveal that materials for adsorbing biological components with a halogen content per gram of dry weight of water-insoluble carrier below a certain level have a low albumin adsorption rate and suppress nonspecific adsorption of proteins.

[0144] Furthermore, it was revealed that a biological component adsorption material having an amino group content of 0.12 mmol or more per gram of dry weight of the water-insoluble carrier has a high adsorption rate of latent TGF-β and can exhibit adsorption performance for target substances.

[0145] The biological component adsorption material of the present invention can adsorb specific target substances while suppressing non-specific adsorption of proteins, and therefore can be used in blood purification columns for treating diseases in which an increase in pathogenic substances is observed in the blood, and in affinity separation columns for purifying target substances from biological fluids.

Claims

1. A biological component adsorption material comprising a water-insoluble carrier having a ligand having an amino group or a negatively charged functional group, a chemical structure containing a halogen atom, and a substrate, wherein the ligand and the chemical structure are bound to the substrate, and the content of the halogen atom is 0.05 to 2.00 mmol per 1 g of the dry weight of the water-insoluble carrier.

2. The biological component-adsorbing material according to claim 1, wherein the content of the halogen atoms is 0.06 to 1.50 mmol per gram of the dry weight of the water-insoluble carrier.

3. The biological component adsorption material according to claim 1 or 2, wherein the negatively charged functional group is a sulfonic acid group or a carboxyl group.

4. The biological component adsorption material according to any one of claims 1 to 3, wherein the halogen atom is a chlorine atom.

5. A biological component adsorption material according to any one of claims 1 to 4, wherein the content of the amino group or negatively charged functional group is 0.04 to 0.50 mmol per 1 g of the dry weight of the water-insoluble carrier.

6. The biological component adsorption material according to any one of claims 1 to 5, wherein the ligand is a ligand having an ethyleneimine polymer.

7. The biological component adsorption material according to any one of claims 1 to 5, wherein the negatively charged functional group is a thioalkylsulfonic acid group or a thioalkylcarboxyl group.

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