Adsorption materials and adsorption columns

Abstract

To provide an adsorption material capable of selectively adsorbing immune suppression leucocytes.SOLUTION: A cell adsorption material includes a water insoluble carrier in which one or more kinds of nitrogen-containing compounds selected from a group formed of polyamine and aliphatic amine are coupled, a shape of the water insoluble carrier is a fiber or particle shape, a long diameter of the fiber or the particle is 15 μm or greater and 50 μm or smaller, and when the water insoluble carrier is immersed in a physiologic saline (hereafter, called as filling liquid) and is subjected to steam sterilization, an amount of an effluent in the filling liquid after performing the steam sterilization is 60 μg / g or smaller.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present disclosure relates to adsorption materials and adsorption columns for cells, such as LAP-positive immune cells. [Background technology]

[0002] It has become clear that cancer is closely related to the immune system, and in recent years, it has been reported that immunosuppressive blood components are elevated in many advanced cancers. One of these blood components is white blood cells, which are classified into lymphocytes, granulocytes, and monocytes. Each white blood cell is further subdivided; for example, lymphocytes are classified into T cells, B cells, natural killer cells, etc.

[0003] Cancer cells are known to have a mechanism for evading the immune system by producing immunosuppressive substances, such as transforming growth factor-β (TGF-β), which induce immunosuppressive T cells. TGF-β is produced as a latent LAP-TGF-β complex, which is non-covalently associated with Latency Associated Peptide (LAP), which has a molecular weight of 75,000. When LAP is cleaved by proteolytic enzymes or cell adhesion molecules, TGF-β is able to bind to TGF-β receptors on various cells and exert its physiological functions.

[0004] It is known that latent LAP-TGF-β complexes bind not only to cancer cells but also to the cell membranes of some immune cells. Immune cells to which this LAP-TGF-β complex binds are called LAP-positive immune cells, and this is one of the reasons why cancer cells escape from the immune system. In particular, among the LAP-positive immune cells, LAP-positive T cells have attracted attention as a source of TGF-β for cancer cells, and efforts have been made to develop adsorbents to remove them.

[0005] On the other hand, platelets are also known to be a source of TGF-β, so if it were possible to simultaneously remove LAP-positive immune cells, including LAP-positive T cells and LAP-positive platelets, which are sources of the immunosuppressant TGF-β, it is expected that the effectiveness of cancer treatment would be improved.

[0006] As a method of treating cancer, drugs such as immune checkpoint antibodies have been developed that inhibit the transmission of immunosuppressive signals emitted by cancer cells, but there have also been cases where patients have developed autoimmune diseases due to side effects of these drugs.

[0007] Additionally, cell therapy, in which the patient's own white blood cells eliminate cancer cells, is being used to improve immune function while reducing side effects. A typical method is dendritic cell infusion therapy, in which the patient's dendritic cells are loaded with cancer antigens ex vivo and then returned to the patient to induce cancer-specific killer T cells for treatment. However, at present, this treatment is considered to be insufficiently effective. One of the reasons for this is thought to be the enhancement of the immune suppression system.

[0008] On the other hand, if it were possible to remove LAP-positive immune cells, which are the source of TGF-β, it is expected that the immune response against cancer cells would be maintained, making it possible to kill cancer cells and suppress the progression of cancer.

[0009] Patent Document 1 discloses a filter that is characterized by the fiber diameter and bulk density of the nonwoven fabric as a material for removing leukocytes. Specifically, the fiber diameter is less than 3 μm and the bulk density is 0.15 g / cm. 3 Exceeds 0.50g / cm 3 The following nonwoven fabric leukocyte removal filter is disclosed.

[0010] Patent Document 2 discloses an adsorbent material characterized by fiber diameter, surface area, etc., and specifically, the material contains fibers with a fiber diameter of 0.5 to 10 μm and a surface area of ​​0.5 m 2 More than 10m 2The present invention discloses a cell adsorption column packed with an adsorbent having a volume of less than 100 ml, characterized in that the adsorbent packed volume is 100 ml or less.

[0011] Patent Document 3 discloses an adsorption material for immunosuppressive leukocytes, characterized in that the arithmetic roughness of the fiber surface is 0.1 to 3.0 μm or less. [Prior art documents] [Patent documents]

[0012] [Patent Document 1] Japanese Patent Application Laid-Open No. 60-193468 [Patent Document 2] WO2008 / 038785 No. Summary of the Invention [Problem to be solved by the invention]

[0013] Conventionally, methods have been used to introduce ligands into water-insoluble adsorbents to improve adsorption selectivity for target substances in liquids. For example, Patent Document 1 discloses a method of introducing amines to the surface of a knitted fabric to remove leukocytes, and Patent Document 2 discloses a method of similarly introducing negatively charged ligands to the surface of a knitted fabric to remove proteins such as cytokines. In these ligand introduction reactions, organic solvents are often used to increase the affinity between the adsorbent and the solvent and introduce more ligands onto the surface of the water-insoluble carrier. Furthermore, amines are sometimes used as catalysts to further improve reaction efficiency. Meanwhile, for medical devices that come into contact with blood, strict regulations are imposed on residual solvents and eluates, and it is desirable to minimize their use. Furthermore, the use of organic solvents can cause the water-insoluble carrier to swell and deform. This can lead to the problem that ligands and catalysts that penetrate the water-insoluble interior due to swelling remain inside the adsorbent carrier after the reaction and may leach into the filler liquid, necessitating improvements.

[0014] Therefore, by changing the reaction solvent from an organic solvent to an aqueous solvent which has a low affinity for the water-insoluble support, swelling of the water-insoluble support is suppressed, and further penetration of the ligand and catalyst into the water-insoluble support is suppressed, resulting in the unexpected effect of dramatically reducing eluates, which led to the present invention. [Means for solving the problem]

[0015] The present invention, which solves the above-mentioned problems, is as follows.

[0016] a water-insoluble carrier having one or more nitrogen-containing compounds bound thereto selected from the group consisting of polyamines and aliphatic amines; the water-insoluble carrier is in the form of fibers or particles, The major axis of the fiber or particle is 15 μm or more and 45 μm or less, A cell adsorption material in which, when the water-insoluble carrier is immersed in physiological saline (hereinafter referred to as filling solution) and steam sterilized, the amount of eluate in the filling solution after steam sterilization is 60 μg / g or less. [Effects of the Invention]

[0017] According to the present disclosure, an adsorption material capable of removing LAP-positive cells with reduced elution when subjected to high-temperature steam sterilization is provided. [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 is a longitudinal cross-sectional view of an example of a radial flow type adsorption column. DETAILED DESCRIPTION OF THE INVENTION

[0019] The present invention provides a cell adsorption material comprising a water-insoluble carrier bound to one or more nitrogen-containing compounds selected from the group consisting of polyamines and aliphatic amines, the water-insoluble carrier having a fibrous or granular shape, the fibers or particles having a major axis of 15 μm or more and 50 μm or less, and when the water-insoluble carrier is immersed in physiological saline (hereinafter referred to as "filling solution") and steam sterilized, the amount of eluate in the filling solution after steam sterilization is 60 μg / g or less.

[0020] The present embodiment will be described in more detail below. It should be noted that, unless otherwise specified, the terms used in this specification are used in the same way as commonly used in the relevant field. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. In the event of any conflict, the present specification (including definitions) shall prevail.

[0021] The adsorptive material according to this embodiment relates to an adsorptive material for cells such as LAP-positive immune cells. LAP-positive immune cells are cells with LAP-TGF-β complexes bound to their cell membrane surfaces. Specific examples of LAP-positive immune cells include LAP-positive T cells and LAP-positive platelets, but the present invention is not limited to these examples as long as the LAP-TGF-β complexes are bound to the cell membrane surface.

[0022] When the water-insoluble carrier of the adsorbent material according to this embodiment is immersed in physiological saline (hereinafter referred to as the "filling solution") and steam sterilized, the amount of eluates in the filling solution after steam sterilization is 60 μg / g or less. In addition to the ligand, the eluates may also contain catalysts if used during the reaction. Examples of catalysts include amines such as triethyleneamine. Steam sterilization is a method of killing microorganisms by heating with saturated steam at an appropriate temperature and pressure in a sealed device. The advantage of steam sterilization is that it can heat rapidly, allowing heat to penetrate deep into the object to be sterilized, thereby reliably killing all microorganisms, including heat-resistant spore-forming bacteria, in a relatively short period of time. Furthermore, steam sterilization is applicable to many instruments, objects, and liquid substances, and is relatively resistant to material deterioration and alteration, making it widely used in medical institutions and pharmaceutical manufacturing facilities. In this specification, steam sterilization refers to heat treatment at 115°C for 30 minutes or more.

[0023] The amount of eluate refers to the amount of eluate in solution measured by oxidative decomposition-reduced pressure chemiluminescence, and in the present invention, this is 60 μg / g or less. To determine the amount of eluate, pyrolysis and oxidation were performed at 800°C, and the generated nitric oxide was measured by chemiluminescence. Quantitative determination was performed by analyzing an aqueous potassium hydroxide solution as a standard solution and calculating the concentration using a calibration curve. Measurements were performed using an ND-100 trace nitrogen analyzer (manufactured by Mitsubishi Chemical Corporation). The measurement conditions for the instrument were: pyrolysis temperature 800°C, catalyst temperature 900°C, main oxygen flow rate 300 mL / min, sub-oxygen flow rate 300 mL / min, Ar flow rate 400 mL / min, and Sens set to High.

[0024] The adsorption material according to this embodiment includes a water-insoluble carrier to which one or more nitrogen-containing compounds selected from the group consisting of polyamines and aliphatic amines are bonded. The nitrogen-containing compound bonded to the water-insoluble carrier is not particularly limited as long as it is a polyamine or an aliphatic amine, and details will be described later.

[0025] The adsorptive material of this embodiment can selectively adsorb cells, particularly LAP-positive immune cells, and preferably has an excellent adsorption rate for LAP-positive immune cells.

[0026] The nitrogen-containing compound bound to the water-insoluble carrier is selected from polyamines and aliphatic amines. The nitrogen-containing compound may be used alone or in combination. The nitrogen-containing compound may be bound directly or indirectly to the water-insoluble carrier. The group obtained by directly or indirectly binding the nitrogen-containing compound to the water-insoluble carrier is referred to herein as a "nitrogen-containing compound residue."

[0027] In one embodiment, the nitrogen-containing compound is a polyamine represented by formula (3).

[0028] R 1 R 2 NX-NR 3 R 4 ...Equation (3) [In formula (3), X represents a saturated or unsaturated aliphatic hydrocarbon group having 2 to 20 carbon atoms, or a heteroatom-containing carbon chain in which 1 to 5 carbon atoms in a saturated or unsaturated aliphatic hydrocarbon group having 3 to 20 carbon atoms are replaced with nitrogen atoms, and the hydrogen atoms bonded to the nitrogen atoms may be substituted with alkyl groups which may have an amino group. R 1 ~R 4 are each independently a hydrogen atom or an alkyl group. In formula (3), X is, for example, a saturated or unsaturated aliphatic hydrocarbon group having 2 to 20 carbon atoms (e.g., 16 or less, 14 or less, 12 or less, 10 or less, 8 or less, 6 or less, 4 or less). In formula (3), X is, for example, a heteroatom-containing carbon chain in which 1 to 5 (e.g., 1 to 3) carbon atoms in a saturated or unsaturated aliphatic hydrocarbon group having 3 to 20 carbon atoms (e.g., 16 or less, 14 or less, 12 or less, 10 or less) are replaced with nitrogen atoms, and the hydrogen atoms bonded to the nitrogen atoms may be substituted with an alkyl group (e.g., an alkyl group having 1 to 6 (preferably 1 to 4) carbon atoms) that may have an amino group. And R 1 ~R 4 are each independently a hydrogen atom or an alkyl group. The alkyl group has, for example, 1 to 6 (preferably 1 to 4) carbon atoms. The aliphatic hydrocarbon group may be linear or branched.

[0029] The polyamine represented by formula (3) is preferably a polyamine represented by any one of the following formulas (3-1) to (3-6).

[0030] H2N-(CH2) p1 -NH2...Formula (3-1) [In formula (3-1), p1 is an integer of 2 to 12 (preferably 2 to 6, 2 to 5, or 2 to 4), and at least one of the hydrogen atoms of the primary amino groups at both ends may be substituted with an alkyl group.] H2N-(CH2) p1 -NH-(CH2) p2 -NH2...Formula (3-2) [In formula (3-2), p1 and p2 each independently represent an integer of 2 to 5 (preferably 2 to 4, 2 to 3, or 2), and the hydrogen atom of the secondary amino group may be substituted with an alkyl group which may have an amino group, and at least one of the hydrogen atoms of the primary amino groups at both ends may be substituted with an alkyl group.] H2N-(CH2) p1 -NH-(CH2) p2 -NH-(CH2) p3 -NH2...Formula (3-3) [In formula (3-3), p1, p2, and p3 each independently represent an integer of 2 to 5 (preferably 2 to 4, 2 to 3, 2), and the hydrogen atoms of the secondary amino groups may each independently be substituted with an alkyl group which may have an amino group, and at least one of the hydrogen atoms of the primary amino groups at both ends may be substituted with an alkyl group.] H2N-(CH2) p1 -NH-(CH2) p2 -NH-(CH2) p3 -NH-(CH2) p4 -NH2...Formula (3-4) [In formula (3-4), p1, p2, p3, and p4 each independently represent an integer of 2 to 5 (preferably 2 to 4, 2 to 3, 2), the sum of p1, p2, p3, and p4 is 17 or less, the hydrogen atoms of the secondary amino groups may each independently be substituted with an alkyl group which may have an amino group, and at least one of the hydrogen atoms of the primary amino groups at both ends may be substituted with an alkyl group.] H2N-(CH2) p1 -NH-(CH2) p2 -NH-(CH2) p3 -NH-(CH2) p4 -NH-(CH2) p5 -NH2...Formula (3-5) [In formula (3-5), p1, p2, p3, p4, and p5 each independently represent an integer of 2 to 5 (preferably 2 to 4, 2 to 3, or 2), the sum of p1, p2, p3, p4, and p5 is 16 or less, the hydrogen atoms of the secondary amino groups may each independently be substituted with an alkyl group which may have an amino group, and at least one of the hydrogen atoms of the primary amino groups at both ends may be substituted with an alkyl group.] H2N-(CH2) p1 -NH-(CH2) p2 -NH-(CH2) p3 -NH-(CH2) p4 -NH-(CH2) p5 -NH-(CH2) p6 -NH2...Formula (3-6) [In formula (3-6), p1, p2, p3, p4, p5, and p6 each independently represent an integer of 2 to 5 (preferably 2 to 4, 2 to 3, or 2), the sum of p1, p2, p3, p4, p5, and p6 is 15 or less, the hydrogen atoms of the secondary amino groups may each independently be substituted with an alkyl group which may have an amino group, and at least one of the hydrogen atoms of the primary amino groups at both ends may be substituted with an alkyl group.]

[0031] In formulas (3-2) to (3-6), the number of carbon atoms in the "alkyl group which may have an amino group" which can be bonded to the nitrogen atom of the secondary amino group is, for example, 1 to 6, preferably 1 to 5, more preferably 1 to 4, and even more preferably 1 to 3. In formulas (3-1) to (3-6), the number of carbon atoms in the "alkyl group" which can be bonded to the nitrogen atoms of the primary amino groups at both ends is, for example, 1 to 6, preferably 1 to 5, preferably 1 to 4, and preferably 1 to 3. These "alkyl groups" are preferably linear or branched.

[0032] Examples of polyamines represented by formula (3) suitable as nitrogen-containing compounds include ethylenediamine, N-ethylethylenediamine, diethylenetriamine, N-ethyldiethylenetriamine, triethylenetetraamine, and tetraethylenepentamine. Other examples include the following polyamines: 3,3'-diaminodipropylamine, 1,3-diaminopropane, norspermidine, homospermidine, aminopropylcadaverine, aminobutylcadaverine, norspermine, thermospermine, aminopropylhomospermidine, canavalmine, homospermine, aminopentylnorspermidine, N,N-bis(aminopropyl)cadaverine, caldopentamine, homocaldopentamine, thermopentamine, caldohexamine, homocaldohexamine, thermohexamine, homothermohexamine, N 4 -aminopropylnorspermidine; N 4 -aminopropylspermidine; N 4 -aminopropylnorspermine.

[0033] In the adsorbent material of the present invention, when the nitrogen-containing compound is an aliphatic amine, in one embodiment, the nitrogen-containing compound is preferably a primary aliphatic amine represented by formula (4) or a secondary aliphatic amine represented by formula (5).

[0034] NH2R 5 ...Equation (4) [In formula (4), R 5 is a saturated or unsaturated aliphatic hydrocarbon group having 1 to 12 carbon atoms. NHR 6 R 7 ...Equation (5) [In formula (5), R 6 and R 7 are each independently a saturated or unsaturated aliphatic hydrocarbon group having 1 to 12 carbon atoms.

[0035] In the aliphatic amine represented by formula (4) or (5), the aliphatic hydrocarbon group is preferably a linear or branched aliphatic hydrocarbon group, and more preferably a linear or branched saturated aliphatic hydrocarbon group. The aliphatic hydrocarbon group preferably has 1 to 8 carbon atoms, more preferably 1 to 6 carbon atoms, and even more preferably 1 to 4 carbon atoms.

[0036] Specific examples of aliphatic amines suitable as the nitrogen-containing compound include monoalkylamines such as ethylamine, propylamine, butylamine, pentylamine, hexylamine, heptylamine, octylamine, nonylamine, and decylamine; and dialkylamines such as diethylamine, dipropylamine, dibutylamine, diheptylamine, dioctylamine, and dicyclohexylamine.

[0037] Other suitable nitrogen-containing compounds include, for example, ethylamine, ethylenediamine, diethylamine, N-ethylethylenediamine, diethylenetriamine, N-ethyldiethylenetriamine, triethylenetetraamine, and tetraethylenepentamine. Among these, ethylenediamine, diethylenetriamine, triethylenetetraamine, and tetraethylenepentamine are preferred. The nitrogen-containing compounds are commercially available or can be produced by known methods or methods similar thereto.

[0038] The water-insoluble carrier having a nitrogen-containing compound bonded thereto includes both a water-insoluble carrier having a nitrogen-containing compound bonded directly and covalently, and a water-insoluble carrier having a nitrogen-containing compound bonded indirectly via a linker. The water-insoluble carrier having a nitrogen-containing compound bonded thereto also includes a water-insoluble carrier having two or more different nitrogen-containing compounds bonded thereto.

[0039] When a polyamine represented by formula (3) is used as the nitrogen-containing compound, multiple amino groups may be bound to the water-insoluble support to form a crosslinked structure. That is, when a polyamine represented by formula (3) is bound to a water-insoluble support as the nitrogen-containing compound, a crosslinked structure is formed when at least two of the amino groups in the polyamine are bound to the water-insoluble support.

[0040] The nitrogen-containing compound is preferably bound to the water-insoluble carrier via an amino group (or nitrogen atom) in the compound.

[0041] When a nitrogen-containing compound binds to a water-insoluble support, a primary amino group, a secondary amino group, a tertiary amino group, and / or a quaternary amino group will be present in the nitrogen-containing compound after binding, depending on the binding position in the nitrogen-containing compound. For example, when a polyamine represented by formula (3) binds to a water-insoluble support, a primary amino group, a secondary amino group, a tertiary amino group, and / or a quaternary amino group will be present in the polyamine after binding, depending on the binding position in the polyamine. Furthermore, the nitrogen-containing compound is preferably bound to the water-insoluble support via an amino group (or nitrogen atom) in the compound. In this specification, the term "amino group in the water-insoluble support" refers to at least the primary amino group, secondary amino group, tertiary amino group, and quaternary amino group derived from the nitrogen-containing compound thus generated. When the nitrogen-containing compound is bound to the water-insoluble support via a linker, the term "amino group in the water-insoluble support" refers to the primary amino group, secondary amino group, tertiary amino group, and quaternary amino group derived from the linker. In addition, in this specification, the "total amount of amino groups" means the total amount (µmol) of primary amino groups, secondary amino groups, tertiary amino groups, and quaternary amino groups in the water-insoluble carrier.

[0042] The total amount of amino groups in the water-insoluble carrier is not particularly limited, but is, for example, 20 μmol or more and 200 μmol or less per 1 g of the adsorption material.

[0043] The total amount of amino groups in a water-insoluble carrier can be determined, for example, by measuring the amino groups using acid-base back titration, as the sum of the amount of primary amino groups, secondary amino groups, tertiary amino groups, and quaternary amino groups (quaternary ammonium groups). Specifically, first, the adsorbent material and an excess amount of aqueous sodium hydroxide solution are added to a polypropylene container and thoroughly stirred at room temperature to desalt the salt-attached amino groups in the adsorbent material. Next, the adsorbent material is thoroughly washed with ion-exchanged water until the solution becomes neutral, and then dried until the weight change is 1% or less. Next, the amino groups in the dried adsorbent material are reacted with a certain amount of a standard solution containing an excess of acid. The amount of acid remaining unreacted with the amino groups is then titrated with a standard solution containing a base. This method allows the total amount of amino groups (μmol) to be determined. More specifically, the total amount of amino groups in a water-insoluble carrier can be determined by the method described in the Examples below. Although this method can measure the amount of quaternary amino groups, in this example, quaternary amino groups are not contained in the structure of the nitrogen-containing compound used, and the amount is the sum of the amount of primary amino groups, the amount of secondary amino groups, and the amount of tertiary amino groups.

[0044] The amount of nitrogen-containing compound residues in the water-insoluble carrier (immobilized amount of nitrogen-containing compound) can be controlled, for example, by adjusting the amount of reactive functional groups bound to the water-insoluble carrier, the type of nitrogen-containing compound, and the amount of nitrogen-containing compound used. The amount of reactive functional groups bound can be controlled, for example, by reaction conditions such as the type of reactive functional group or type of solvent, immersion temperature, or immersion time. For example, when the water-insoluble carrier contains polyaromatic vinyl, a crosslinking agent can also be used to control the binding sites of the reactive functional groups. Furthermore, the amount of nitrogen-containing compound residues can be controlled by reaction conditions such as the type of solvent, immersion temperature, and immersion time, in addition to the type of nitrogen-containing compound and the amount of reactive functional groups bound.

[0045] A "water-insoluble carrier" refers to a carrier that does not change in weight when immersed in water at room temperature (25°C). Specifically, it is preferable that the carrier undergoes a weight change of 5% or less when immersed in water at 25°C for 1 hour.

[0046] The material for the water-insoluble carrier is not particularly limited, but preferred examples include polyaromatic vinyl compounds such as polystyrene, polyethersulfone, polysulfone, polyarylethersulfone, polyetherimide, polyimide, polyamide, and polyphenylene sulfide. Water-insoluble carrier materials are commercially available or can be produced by known methods or methods similar thereto. These materials are substantially free of hydroxyl groups, which are known to easily activate complement upon contact with blood. Among these, polystyrene is preferred as the material for the water-insoluble carrier because it has a large number of aromatic rings per unit weight and is easy to immobilize amino groups. These polymer materials may be used alone or in combination. Furthermore, the water-insoluble carrier is preferably a polymer material containing a polyaromatic vinyl compound (e.g., polystyrene). As the water-insoluble carrier, a copolymer of polystyrene and polyolefin (e.g., a copolymer of polystyrene and polyethylene or a copolymer of polystyrene and polypropylene) is preferred because it is easy to introduce a linker such as an active halogen group to fix an amino group to the polystyrene portion, and because the strength reinforcement provided by the polyolefin portion makes it easy to handle and chemically resistant. The polymer material may also be a blend or alloy, and a polymer alloy of polystyrene and polyolefin (e.g., a polymer alloy of polystyrene and polyethylene or a polymer alloy of polystyrene and polypropylene) is particularly preferred because it has chemical resistance and easily retains its physical shape. Among these, a polymer alloy of polystyrene and polypropylene, which has a proven track record in extracorporeal blood circulation therapy, is preferred. The water-insoluble carrier used preferably has substantially no amino groups.

[0047] The nitrogen-containing compound may be directly bound to the water-insoluble carrier, or may be indirectly bound to the water-insoluble carrier via a linker. The method for binding the nitrogen-containing compound to the water-insoluble carrier is not particularly limited, and examples thereof include a method in which the nitrogen-containing compound is covalently bound to the surface of the water-insoluble carrier via a linker by a chemical method.

[0048] When the nitrogen-containing compound and the water-insoluble carrier are directly covalently bonded, the covalent bond preferably has an electrically neutral chemical bond such as an amide bond, a urea bond, an ether bond, or an ester bond, and more preferably an amide bond or a urea bond. When the nitrogen-containing compound is bonded to the water-insoluble carrier via a linker, that is, when the nitrogen-containing compound is bonded to the linker by the above-mentioned covalent bond, it is preferable that the linker before bonding the nitrogen-containing compound to the water-insoluble carrier via the linker (i.e., the linker before reaction) has a reactive functional group. Examples of the reactive functional group possessed by the linker before reaction include active halogen groups such as halomethyl groups, haloacetyl groups, haloacetamidomethyl groups, or halogenated alkyl groups, epoxy groups, carboxyl groups, isocyanate groups, thioisocyanate groups, or acid anhydride groups. Among these, active halogen groups (especially haloacetyl groups) are preferred because they are easy to produce, have moderately high reactivity, can be used to carry out the immobilization reaction of amino groups under mild conditions, and the resulting covalent bond is chemically stable. Specific examples of polymers with reactive functional groups include polystyrene with chloroacetamidomethyl groups added, polysulfone with chloroacetamidomethyl groups added, and polyetherimide with chloroacetamidomethyl groups added. These polymers are soluble in organic solvents and have the advantage of being easily moldable. The amount of linker introduced depends on the structure of the linker, but for example, in the case of an amidomethyl group, it is preferably 1 mmol to 10 mmol per 1 g of water-insoluble carrier.

[0049] In this embodiment, the amino group / amide group ratio in the water-insoluble carrier is preferably 0.01 or more and 0.06 or less. If it is less than 0.01, the amount of nitrogen-containing compound bound to the water-insoluble carrier is small, which may reduce cell removal performance. On the other hand, if it exceeds 0.06, monocytes and granulocytes, among white blood cells, may be activated and adsorbed to the water-insoluble carrier in large amounts, which may inhibit the selective removal of cells such as LAP-positive immune cells. In addition, there is a possibility that adsorption of anticoagulants such as heparin may increase.

[0050] The reactive functional group can be introduced by reacting it with a water-insoluble support in advance. For example, if the water-insoluble support is polystyrene and the reactive functional group is a chloroacetamidomethyl group, polystyrene with a chloroacetamidomethyl group can be obtained by reacting polystyrene with N-methylol-α-chloroacetamide.

[0051] The water-insoluble carrier may be in the form of fibers or particles. Fibers are particularly preferred because they can be processed to ensure a blood flow path and increase the contact area with blood.

[0052] When the water-insoluble carrier has a fibrous shape, it is preferably an islands-in-sea type composite fiber. From the viewpoint of maintaining the strength of the material, an islands-in-sea type composite fiber in which the islands are a reinforcing material and the sea is an alloy of a water-insoluble polymer and a reinforcing material is preferred, and an islands-in-sea type composite fiber in which the islands are polypropylene and the sea is an alloy of polystyrene and polypropylene is even more preferred. The reinforcing material is not particularly limited, but examples include polyamide, polyacrylonitrile, polyethylene, polypropylene, nylon, polymethyl methacrylate, polytetrafluoroethylene, etc. Among these, polypropylene is preferred. These polymers may be used alone or in combination.

[0053] When the water-insoluble carrier is in the form of fiber, it is preferably in the form of a knitted fabric, which is an advanced processed product thereof. By controlling the stitches of the knitted fabric, blood flow paths can be secured, thereby reducing pressure loss when blood passes through the fibers. Furthermore, when the knitted fabric is formed by doubling fibers, the number of doubling yarns is preferably 10 to 100, and more preferably 30 to 80. When the number of doubling yarns is 100 or less, LAP-positive immune cells can easily enter the fiber bundle, improving the adsorption rate. Furthermore, when the number of doubling yarns is 10 or more, the shape retention of the knitted fabric is improved. Any of the preferred lower limits can be combined with any of the preferred upper limits.

[0054] In this embodiment, the water-insoluble carrier has the form of fibers or particles. The major axis of the fibers or particles is not particularly limited, but is preferably 15 μm or more and 50 μm or less.

[0055] In this embodiment, when the major axis of the fibers or particles constituting the water-insoluble carrier is within the range of 15 μm to 50 μm, the blood contact area per unit volume of the water-insoluble carrier can be increased while maintaining an appropriate packing density of the water-insoluble carrier in the column. This is presumably to suppress non-selective adsorption of platelets, leukocytes, etc., and to increase the adsorption rate of LAP-positive immune cells. Therefore, when the water-insoluble carrier is in the form of fibers or particles, the major axis of the fibers or particles is preferably 15 μm to 50 μm. However, these presumptions do not limit the present embodiment. When the water-insoluble carrier is in the form of fibers or particles, the major axis is preferably 17 μm or more, 20 μm or more, or 25 μm or more. The major axis of the fibers or particles is preferably 40 μm or less, 35 μm or less, or 30 μm or less. Any preferred lower limit can be combined with any preferred upper limit.

[0056] The "long diameter of the fiber" can be determined by the following method. First, 100 fiber samples are randomly collected, and a photograph of the cross section (a cross section perpendicular to the elongation direction of the fiber) is taken for each sample using a scanning electron microscope at a magnification of 1000 to 3000. Next, the long diameter of each fiber cross section is measured. The average of these values ​​(the average of the long diameters of a total of 100 fiber cross sections) is calculated to determine the "long diameter of the fiber."

[0057] The "major diameter of the particle" can be determined by the following method. First, ten particle samples are randomly collected, and one photograph is taken for each sample using a scanning electron microscope at a magnification of 1000 to 3000. Next, the major diameters of ten particles per photograph are measured. The average of these values ​​(the average of the major diameters of a total of 100 particles) is then calculated to determine the "major diameter of the particle."

[0058] In this embodiment, the cross-sectional circularity of the water-insoluble carrier is not particularly limited, but is preferably 0.70 or more and 0.95 or less. The cross-sectional circularity of the water-insoluble carrier is preferably 0.75 or more, and 0.80 or more. The upper limit of the cross-sectional circularity of the water-insoluble carrier is 0.95 or less, and preferably 0.90 or less. Any preferred lower limit can be combined with any preferred upper limit. When the cross-sectional circularity is in the range of 0.70 or more and 0.95 or less, it is presumed that nonspecific adsorption of monocytes and granulocytes to the water-insoluble carrier is suppressed, and LAP-positive immune cells are selectively and efficiently removed.

[0059] "Circularity" is a numerical value that indicates how much a target circular shape deviates from a perfect circle, and is calculated using the following formula (6). Note that circularity takes a value between 0 and 1, and the closer the value is to 1, the closer the target circular shape is to a perfect circle.

[0060] 4π×S / a 2 ...Equation (6) [In formula (6), S is the area of ​​the target circular shape, and a is the perimeter of the target circular shape.] The "circularity of the water-insoluble carrier" is defined as "the circularity of the water-insoluble carrier in the region A and the region B." 26 CN - The "spectral intensity" of the sample is calculated from the image Y acquired at the time of measurement using the image analysis software Avizo and ImageJ.

[0061] In this embodiment, the arithmetic mean roughness of the surface of the water-insoluble carrier is preferably 0.01 μm or more and 3.0 μm or less.

[0062] The "arithmetic mean roughness" is a value calculated by the following formula (7) when a reference length L is extracted from a roughness curve 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). This value means the arithmetic mean roughness (Ra) of JIS B 0601-2001. The arithmetic mean roughness can be measured, for example, with a shape measurement laser microscope. The measurement is preferably performed while the water-insoluble carrier is dry. In addition, when there is orientation, such as with fibers, the value in the longitudinal direction is measured.

[0063]

number

[0064] The arithmetic mean roughness of the surface of the water-insoluble carrier is more preferably 0.05 μm or more and 3.0 μm or less. It is estimated that when the arithmetic mean roughness of the surface of the water-insoluble carrier is in the range of 0.01 μm or more and 3.0 μm or less, it is possible to maintain the adsorption performance of LAP-positive cells and suppress nonspecific adsorption of monocytes and granulocytes. Therefore, the arithmetic mean roughness of the surface of the water-insoluble carrier is preferably 0.05 μm or more, 0.1 μm or more, 0.2 μm or more, 0.3 μm or more, and 0.4 μm or more in this order. The arithmetic mean roughness of the surface of the water-insoluble carrier is preferably 3.0 μm or less, more preferably 0.9 μm or less, and even more preferably 0.8 μm or less. Any preferred lower limit can be combined with any preferred upper limit. The preferred major diameter of the fibers or particles of the water-insoluble carrier, the preferred circularity of the water-insoluble carrier, and the preferred arithmetic mean roughness of the surface of the water-insoluble carrier can be combined in any manner.

[0065] The arithmetic mean roughness of the surface of the water-insoluble carrier can be controlled, for example, by immersing the water-insoluble carrier in an organic solvent. As a method for controlling the arithmetic mean roughness of the surface of the water-insoluble carrier, for example, a method can be mentioned in which a polymer obtained by kneading a polyaromatic vinyl compound and polypropylene as a water-insoluble carrier is partially dissolved in the polyaromatic vinyl compound and then immersed in a solvent that does not dissolve polypropylene. The arithmetic mean roughness of the surface of the water-insoluble carrier can be controlled by the type of polymer, the molecular weight of the polymer, the type of solvent, the immersion temperature, the immersion time, etc. Furthermore, for polyaromatic vinyl, a method can also be adopted in which the solubility in the solvent is controlled by introducing a crosslinking agent. Furthermore, the above reaction can also be carried out simultaneously with the introduction reaction of a nitrogen-containing compound.

[0066] The adsorption material according to the present embodiment can be suitably used when the cells are immune cells, and particularly when the immune cells are LAP-positive immune cells. Among LAP-positive immune cells, the adsorption material can preferably adsorb LAP-positive T cells or LAP-positive platelets, and can preferably adsorb LAP-positive CD4-positive T cells or LAP-positive CD42b-positive platelets.

[0067] In the adsorption material of this embodiment, the adsorption rate of LAP-positive T cells is preferably 70% or more, preferably 80% or more, and preferably 90% or more. The adsorption rate of LAP-positive platelets is preferably 40% or more, preferably 50% or more, more preferably 60% or more, and most preferably 70% or more. In terms of selectivity, the adsorption rates of LAP-positive T cells and LAP-positive platelets are preferably 2.5 times or more, preferably 5 times or more, and preferably 8 times or more than the adsorption rates of LAP-negative T cells and LAP-negative platelets, respectively. Here, CD4-positive T cells are targeted as LAP-positive T cells, and CD42b-positive platelets are targeted as platelets. An example of a test system for the adsorption rate is an adsorption column flow-through immune cell adsorption test using human blood (see, for example, this Example). An example of an evaluation system is flow cytometry analysis using surface antigens of immune cells as indicators (see, for example, this Example).

[0068] The adsorption column of this embodiment includes the adsorption material of this embodiment.

[0069] The term "adsorption column" refers to a column having at least a blood inlet, a case, and a blood outlet, with the case filled with an adsorption material. Examples of adsorption columns include radial flow adsorption columns. As mentioned above, the adsorption material is preferably in the form of fiber, and more preferably knitted.

[0070] An example of the internal configuration of an adsorption column 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 its front and rear longitudinal ends. 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. Of the two partition plates 5 and 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 provided intermittently around the periphery of the partition plate 7. A pipe 8 spans between the opening 5a of the partition plate 5 and the support protrusion 7a of the partition plate 7. The pipe 8 forms a flow path 9 inside for guiding blood, and has a number of through-holes 10 in its peripheral wall. The front end of the pipe 8 is connected to the opening 5a of the partition plate 5, and its rear end is closed by the support protrusion 7a of the partition plate 7. The adsorption material 11 is wrapped around the circumference of the pipe 8 in multiple layers. When this adsorption column is used for the circulation method, a tube forming a circulation circuit between the inlet 2 and the outlet 3 is connected to the blood pool. Blood drawn from the blood pool is supplied to the inlet 2, and the target adsorbed substance (LAP-positive immune cells) is removed by the adsorption material 11 inside the inlet 2, and the blood flows out of the outlet 3 and is returned to the blood pool. Inside the column, blood enters the flow path 9 through the filter 4 from the inlet 2. As it moves through the flow path 9, it gradually penetrates the adsorption material 11 through the through-holes 10 and adsorbs cells and other substances as it moves radially. The blood from which cells and other substances have been removed flows out of the numerous through-holes 7b on the outer periphery of the partition plate 7, passes through the filter 6, and flows out of the 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 the adsorption column may be reversed so that blood is supplied from outlet 3 and flows out from inlet 2.

[0071] To increase the adsorption rate of LAP-positive immune cells, the blood linear velocity in the column is also important. That is, if the blood linear velocity is high, it may be difficult for LAP-positive immune cells to fully interact with the adsorption material. On the other hand, if the blood linear velocity is low, other blood components such as platelets and proteins may adhere nonspecifically to the adsorption material, inhibiting the interaction between the adsorption material and LAP-positive immune cells. Therefore, the flow velocity S at the inlet of the adsorption column is important. in is 50cm 3 The maximum linear velocity of blood in the adsorption material when the flow rate at the inlet of the adsorption column is 50 cm / min or less is preferably 50 cm / min or less, and more preferably 25 cm / min or less. 3 The minimum value of the blood linear velocity in the adsorption material when the blood flow rate is 0.1 cm / min or more is preferably 0.1 cm / min or more, and more preferably 0.5 cm / min or more. Here, the blood linear velocity is determined by calculation. For example, in the case of the following radial flow type adsorption column, the maximum value of the blood linear velocity in the adsorption material (V max ) is the total area of ​​the openings on the sides of the central pipe (S p ) and the flow rate S at the inlet of the adsorption column in (50cm 3 / min) using the following formula (8).

[0072] V max (cm / min)=S in (cm 3 / min) / S p (cm 2 )...Equation (8) Also, the minimum value (V min ) is the area of ​​the outermost surface of the adsorption material wrapped around the central pipe (S o ) and the flow rate S at the inlet of the adsorption column in (50cm 3 / min) using the following formula (9): min (cm / min)=S in (cm 3 / min) / S o (cm 2 )...Equation (9) On the other hand, when the shape of the adsorption material is particles or a fiber shape in which fibers are simply stacked, the maximum value and the minimum value are the same value.

[0073] Furthermore, a preferred adsorption column is a radial flow type adsorption column comprising a central pipe with through-holes on its longitudinal side for allowing the supplied blood to flow out, an adsorption material filled around the central pipe, and a plate A connected to the upstream end of the central pipe so that the inflowing blood passes through the central pipe and prevents the blood from contacting the adsorption material without passing through the central pipe, and a plate B arranged to block the downstream end of the central pipe and fix the adsorption material in the space around the central pipe. This is to ensure that the blood flows uniformly through the adsorption material. Note that if the through-holes in the central pipe have a low aperture ratio, pressure loss is likely to occur in this area, activating granulocytes, monocytes, and platelets, which then tend to nonspecifically adhere to the adsorption material. This may result in reduced adsorption selectivity for LAP-positive immune cells. Furthermore, if the aperture ratio is high, issues such as reduced pipe strength and increased susceptibility to short-path flow at the through-holes near the blood inlet may arise. Therefore, the aperture ratio of the through holes is preferably 20 to 80%, and more preferably 30 to 60%.

[0074] "Radial flow" refers to the way blood flows inside the column. When blood flows vertically at the inlet and outlet of the column, and there is horizontal blood flow inside the column, it is called radial flow.

[0075] The "opening ratio of the through holes" means the value calculated by the following formula (10).

[0076] Opening rate of through holes (%) = Sum of the area of ​​through holes formed on the side surface in the longitudinal direction of the pipe / Area of ​​the side surface of the pipe × 100... Equation (10) The adsorption column of this embodiment can be used in blood purification therapy. By using the adsorption column of this embodiment as a blood purification column, LAP-positive immune cells can be selectively removed from blood. For example, by circulating blood extracorporeally and passing it through the adsorption column of this embodiment, LAP-positive immune cells can be selectively removed from the blood. That is, the adsorption column of this embodiment can be used as a column for extracorporeal circulation. More specifically, the adsorption column of this embodiment can be used in treatment to selectively remove LAP-positive immune cells from the blood of cancer patients. That is, the adsorption column of this embodiment can be used as a column for cancer treatment.

[0077] The adsorption column of this embodiment is suitable for cancer treatment because it can selectively remove LAP-positive immune cells. It can also be used in combination with cell infusion therapy to activate dendritic cells, natural killer cells, etc.

[0078] The present embodiment will be described below with reference to examples, but the present embodiment is not limited to these examples. [Example]

[0079] 1. Measurement of the arithmetic mean roughness of the surface of a water-insoluble support In this example, the arithmetic mean roughness of the surface of the water-insoluble carrier contained in the adsorption material was measured by the following method.

[0080] Using a shape measuring laser microscope (Color 3D Laser Microscope VK-9700, manufactured by Keyence Corporation), the surface of the adsorbent material was observed at 100x magnification while wetted with water to prevent drying, and the arithmetic mean roughness of the surface was measured (in accordance with JIS B 0601-2001). The reference length L was set to 50 μm, and the average of the absolute values ​​of the deviations measured at 10 different positions was taken as the arithmetic mean roughness of the surface. Note that the water-wet state is preferably one in which the moisture content (moisture content = 100 × water weight / material weight) is 20% or more.

[0081] 2. LAP-positive immune cell adsorption test In this example, the adsorption rate of LAP-positive immune cells to the adsorption material was measured by an adsorption column flow-through immune cell adsorption test using human blood, and analysis was performed using flow cytometry (FACSLyric, Becton Dickinson).

[0082] First, the packed section of an adsorption column (10 mm diameter, 14 mm height) was filled with 0.3 g of adsorption material (dry weight) and saline (hereafter referred to as the "developed column") and then autoclaved (117°C, 105 min). This column was primed by passing saline containing 50 units / mL of heparin at 1.54 mL / min for 2.5 minutes. Next, blood drawn from a healthy human volunteer (containing 5 units / mL of heparin) was passed through the column at 0.31 mL / min for 30 minutes, with blood samples collected from the column outlet every 10 minutes. Blood samples were also collected from the column inlet every 10 minutes. The collected blood was dispensed into 5 mL polypropylene tubes (Falcon) in 100 μL aliquots, and various fluorescently labeled antibodies were added to stain immune cells. The fluorescently labeled antibodies used were fluorescein isothiocyanate (FITC)-labeled anti-human CD4 antibody (BioLegend), phycoerythrin (PE)-labeled anti-human CD42b antibody (BioLegend), and allophycocyanin (APC)-labeled anti-human LAP antibody (R&D). A flow cytometer was used to detect the number of LAP-positive immune cells. CD4-positive and LAP-positive cells were used as LAP-positive T cells, and CD42b-positive and LAP-positive cells were used as LAP-positive platelets.

[0083] The adsorption rate of LAP-positive immune cells was calculated using the following formula (11).

[0084] Adsorption rate of LAP-positive immune cells (%) = (1 - number of LAP-positive immune cells in blood with added adsorbent material / number of LAP-positive leukocytes in blood without added adsorbent material) × 100 Equation (11) 3. Measurement of the major axis when the water-insoluble carrier is in the form of fiber The "long diameter of the fiber" was determined by the following method. First, 100 fiber samples were randomly collected, and a photograph of the cross section (cross section perpendicular to the elongation direction of the fiber) was taken for each sample at a magnification of 3000 using a scanning electron microscope. Next, the diameter of each fiber cross section was measured. The "long diameter of the fiber" was determined by calculating the average of these values ​​(average diameter of the cross sections of a total of 100 fibers). If the fiber cross section was not circular, the diameter of a circle having the same area as the cross section was taken as the long diameter of the fiber.

[0085] 4. Preparation of the base fabric (water-insoluble carrier) and intermediate (ligand-bound water-insoluble carrier) (1) Raw knitted fabric 1 and intermediate body 1 A sea-island composite fiber (fiber diameter: 20 μm) was spun using a water-insoluble carrier containing 264 island components made of polypropylene (Prime Polymer Co., Ltd.; J105WT), and a sea component consisting of 90 wt% polystyrene (weight-average molecular weight: 181,000) and 10 wt% polypropylene (Prime Polymer Co., Ltd.; J105WT), with an island-to-sea ratio (weight ratio) of 50:50. Thirty-six of the resulting fibers were combined to form a knitted fabric (hereinafter referred to as "raw knitted fabric 1"). The arithmetic mean roughness of the fiber surface is affected by factors such as the number of islands, the sea-island ratio, and the molecular weights of the polystyrene and polypropylene.

[0086] Paraformaldehyde (PFA) (2 g) was dissolved in a mixed solution of nitrobenzene (20 mL) and sulfuric acid (13 mL) at 10°C (hereinafter referred to as the PFA solution). Furthermore, N-methylol-α-chloroacetamide (47 g) was dissolved in a mixed solution of nitrobenzene (259 mL) and sulfuric acid (169 mL) at 10°C (hereinafter referred to as the NMCA solution). After immersion and stirring for 1 hour, the knitted fabric was removed and washed with an excess amount of nitrobenzene, then substituted and washed with methanol, and finally washed with ion-exchanged water to obtain the α-chloroacetamidomethylated knitted fabric (hereinafter referred to as Intermediate 1). The entire process, from the preparation of the PFA solution to washing the knitted fabric with methanol, was carried out at 15°C or below.

[0087] 5. Steam sterilization method After the ligand immobilization, 3.5 g (absolute dry weight) of the water-insoluble carrier was wrapped around a central pipe with a diameter of 0.8 cm and then inserted into a polypropylene case with a diameter of 2.1 cm and a height of 4.7 cm. After attaching plates to both sides of the column, ultrasonic welding was performed under a pressure of 200 kPa, a trigger pressure of 44 N, and a welding mode time of 1 second, resulting in a priming volume of 13.9 cm. 3 A column containing the water-insoluble carrier was prepared. After washing the column at 63 mL / min for 55 minutes, the column was switched to saline and washed for an additional 30 minutes. The plate was then sealed with a connector to prevent leakage, placed in an autoclave sterilization bag, and heated at 115°C for 105 minutes. The column solution after heating was used as a sample for measuring eluates.

[0088] 6. Measurement of the amount of extractables in the fill solution After steam sterilization, 10 mL of the filling solution was subjected to oxidative decomposition-reduced chemiluminescence analysis using an ND-100 trace nitrogen analyzer (Mitsubishi Chemical Corporation) to measure the amount of eluted nitrogen-containing compounds. The amount was calculated using a calibration curve based on a potassium hydroxide aqueous solution as a standard solution. The instrument's measurement conditions were: pyrolysis temperature 800°C, catalyst temperature 900°C, main oxygen flow rate 300 mL / min, sub-oxygen flow rate 300 mL / min, Ar flow rate 400 mL / min, and Sens set to High.

[0089] 7. Preparation of Adsorbent Materials Example 1 Diethylenetriamine (hereinafter referred to as DETA) (0.9 mL) was dissolved in ion-exchanged water (427 mL) to prepare a 20 mM DETA solution. Intermediate 1 (10 g) was immersed in this DETA solution and stirred at 40 °C for 3 hours. The product was then immersed in methanol and ion-exchanged water, washed, and dried to obtain adsorption material E1.

[0090] Example 2 DETA (23.3 mL) was dissolved in ion-exchanged water (405 mL) to prepare a 500 mM DETA solution. Intermediate 1 (10 g) was immersed in this DETA solution and stirred at 40 °C for 3 hours. The product was then immersed in methanol and ion-exchanged water, washed, and dried to obtain adsorption material E2.

[0091] Example 3 DETA (46.5 mL) was dissolved in ion-exchanged water (382 mL) to prepare a 1000 mM DETA solution. Intermediate 1 (10 g) was immersed in this DETA solution and stirred at 40 °C for 3 hours. The product was then immersed in methanol and ion-exchanged water, washed, and dried to obtain adsorption material E3.

[0092] (Comparative Example 1) Triethylamine (28.6 mL) was added to dimethyl sulfoxide (DMSO, 398.4 mL), followed by DETA (0.046 mL) to prepare a 1 mM DETA-DMSO solution. Intermediate 1 (10 g) was immersed in this DETA-DMSO solution and stirred at 40°C for 3 hours. The material was then immersed in DMSO, methanol, and water, washed, and dried to obtain adsorbent material E4.

[0093] (Comparative Example 2) The raw knitted fabric 1 was immersed in methanol and ion-exchanged water, washed, and then dried to obtain an adsorbent material E5.

[0094] The arithmetic mean roughness of the surface, the amount of eluate, and the LAP-positive cell removal performance of the adsorbent materials (E1 to E5) were measured by the methods described above. The results are shown in Table 1.

[0095] [Table 1]

[0096] The abbreviations in the table are as follows: DETA: Diethylenetriamine [Industrial Applicability]

[0097] The adsorption material and adsorption column of this embodiment can selectively adsorb cells such as LAP-positive immune cells. Therefore, they are expected to be applied to cancer treatment. In addition, the adsorption material and adsorption column of this embodiment can also be used in combination with cell infusion therapy to activate dendritic cells, natural killer cells, etc. [Explanation of symbols]

[0098] 1 Container body 2 Inlet 3 Outlet 4. Filters 5 Divider 5a Partition opening 6 Filters 7 Divider 7a Partition plate support protrusion 7b Partition plate hole 8 Pipes 9 Flow path 10 through holes 11 Adsorption materials Q Blood flow

Claims

1. A water-insoluble carrier having a polyamine represented by the following formula (3-2) bound thereto, the water-insoluble carrier is in the form of fibers or particles, The major axis of the fiber or particle is 15 μm or more and 50 μm or less, when the water-insoluble carrier is immersed in physiological saline (hereinafter referred to as "filling solution") and steam sterilized, the amount of eluates in the filling solution after steam sterilization is 60 μg / g or less; The arithmetic mean roughness of the surface of the water-insoluble carrier is 0.01 μm or more and 3.0 μm or less. Adsorbed material of LAP-positive T cells or LAP-positive platelets. H 2 N-(CH 2 ) p1 -NH-(CH 2 ) p2 -NH 2 ...Formula (3-2) [In formula (3-2), p1 and p2 each independently represent an integer of 2 to 5.]

2. The adsorption material according to claim 1 , wherein the nitrogen-containing compound is bound to the water-insoluble support via a linker.

3. An adsorption column incorporating the adsorption material according to claim 1 or 2.

4. The adsorption column according to claim 3, which is used in blood purification therapy.

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