JP2015112325A - White blood cell and drug removing method, white blood cell and drug removing filter, and white blood cell and drug removing system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To simultaneously remove at a high rate and in a short time white blood cells and an inactivating drug from a blood product containing red blood cells while suppressing hemolysis.SOLUTION: A white blood cell and drug removing method comprises a process for adding an inactivating drug to a blood product containing red blood cells, introducing the blood product after inactivation into an inlet 2a of a white blood cell and drug removing filter 1 which has a white blood cell removing medium in the upstream side and a drug removing medium in the downstream side, and discharging the blood product from an outlet 2b, to thereby remove the white blood cells in the upstream side and remove the remaining inactivating drug in the downstream side.

Description

  The present invention relates to a filter, a removal system, and a removal method for simultaneously removing leukocytes and a drug from a blood product.

  In recent years, in the field of blood transfusion, a technique for inactivating (inactivating) the proliferating ability of pathogens and leukocytes contained in blood products before blood transfusion has been widespread mainly in Europe. Such an inactivation technique is roughly achieved by drug (inactivation drug) administration, light irradiation, or a combination thereof.

  A pathogen is, for example, a virus, bacterium, protozoan, parasite, or other DNA or RNA. Due to inactivation of pathogens, transfusions such as infections caused by viruses such as HBV, HCV, HIV, HAV, HEV, HTLV, WNV, CMV, infections caused by bacteria, infections caused by protozoa such as Babesia, Chagas disease, and malaria Infections can be prevented. Even if the blood product contains an unknown pathogen species or a pathogen with a dilute concentration level that is difficult to detect, the risk of infection can be suppressed by inactivation. In addition, severe injuries such as post-transfusion graft-versus-host disease (GVHD) caused by the inactivation of leukocytes and the abnormal proliferation of donor lymphocytes in the human body (receiver) who received blood transfusion. Transfusion side effects can be suppressed.

  As inactivation techniques for pathogens and leukocytes, in the case of plasma preparations, S / D method (Otapharma), UVC irradiation (Makopharma), methylene blue and visible light irradiation (Makopharma), amotosalen and UVA irradiation ( Cerus), a combination of riboflavin and UVB irradiation (Thermo BCT).

  In the case of platelet preparations, a combination of amotosalen and UVA irradiation (Cerus) is used mainly in Europe. Furthermore, in the case of erythrocyte preparations, there are those based on ethyleneimine multimers (Inactine, VITEX), acridine derivatives (CERUS), combinations of riboflavin and UVB irradiation (Termo BCT), dimethylmethylene blue (US Red Cross), etc. . Such pathogen inactivation technology itself has been developed.

In the blood product inactivation technique, it is necessary to remove the remaining free drug. This is because the free drug has side effects associated with the inactivation mechanism on the recipient. There are two main inactivation mechanisms. One is an inactivation mechanism in which an organic solvent and a surfactant are added to a blood product to dissolve a phospholipid membrane of a pathogen cell. The other is an inactivation mechanism that prevents transcription and replication of pathogens by adding a chemical binding agent that targets the terminal bases of DNA to blood products and forming an irreversible bond to the DNA bases of pathogens. is there. In either case, if the inactivated residual drug remains in the blood product and is provided for blood transfusion, it will cause damage and antibody production in the recipient's healthy blood cell membrane and somatic cell membrane, causing carcinogenicity and mutagenicity. Since side effects such as showing are known, drug removal is necessary.

  As a technique for removing a drug contained in a blood product, conventionally, as represented by, for example, Patent Document 1, a container having a blood inlet and outlet is filled with porous adsorbent beads, porous fibers, etc. Techniques for controlling physical properties such as fiber diameter and specific surface area have been studied. For example, the above-mentioned porous adsorbent beads and porous fibers are partly contained in a matrix medium, and removed with a batch-type removal technique in which the drug is removed by contact with the blood product for a long time, or the blood product is introduced by a one-pass flow method to remove the drug Flow removal techniques have been established.

On the other hand, in the field of blood transfusion, a method for transfusion of a blood product from which leukocytes have been removed is widely used. This is because, among leukocytes contained in blood products, human leukocyte antigens (HLA) on granulocytes do not match between donors and recipients, or due to invasion of other donor granulocyte antigens. Abnormal reactions such as antibody production and complement activity occur in the body of the blood, and side effects such as fever, vomiting, chills, allergic reactions, granulocytopenia, blood transfusion-related acute lung injury (TRALI), dyspnea, hypotension, and anaphylactic shock This is because there is a risk of causing. In order to reduce these transfusion side effects and perform safer transfusion, it is known that it is necessary to remove until the number of remaining white blood cells in the transfusion preparation unit is 10 ⁴ to 10 ⁶ or less. This means that if there are about 2.0 × 10 ⁹ leukocytes in a standard 400 ml whole blood product, a leukocyte removal rate of at least −3.3 log ₁₀ or higher is required.

  As a technique for removing leukocytes contained in blood products, conventionally, as represented by Patent Document 2, for example, a nonwoven fabric is housed in a container having a blood inlet and outlet, and its fiber diameter, bulk density, bulkiness, surface structure Filtration technology capable of removing leukocytes more easily and efficiently by controlling or combining physical properties such as these has been established.

According to the above prior arts 1 and 2, when both the drug and leukocytes are to be removed from the inactivated blood product, there is no other way but to remove the drug and leukocytes separately. In addition, studies have been made on filters that simultaneously remove leukocytes and drugs.

For example, as in Patent Document 3, a blood product is one-passed against a filter having both a woven filter element with an eyelet bonded with a heparin ligand that adsorbs leukocytes and a filter element containing activated carbon that adsorbs an inactivating agent. A technique for simultaneously removing leukocytes and a drug from a blood product by flowing is disclosed.

Further, as in Patent Document 4, red blood cells were removed by one-pass flow of a blood product from which red blood cells had been removed in advance through a filter provided with non-woven fibers capable of removing white blood cells and activated carbon fibers capable of removing inactivated chemicals. A technique for simultaneously removing leukocytes and an inactivating drug from a blood product is also disclosed.

JP 2010-31049 A Japanese Patent No. 1707022 JP 9-508823 A US Pat. No. 5,667,731

  In the case of the techniques represented by the above-mentioned prior patent documents 1 and 2, it is necessary to separately remove leukocytes and drugs from the blood product. Further, in the case of the techniques represented by the prior patent documents 3 and 4, there is a concept of simultaneously removing leukocytes and drugs from blood products, but the types of blood products actually studied are only plasma and platelets. No means for solving deficiencies in actual use, such as a decrease in flowability due to clogging of the filter or hemolysis, which is a concern when applied to blood products containing red blood cells, is not disclosed. Therefore, it is necessary to apply a filter that simultaneously removes leukocytes and drugs to blood products containing red blood cells without problems, and to examine a method for avoiding clogging, flowability deterioration, and hemolysis of the filter in practical use.

Accordingly, the present invention provides a flow-type leukocyte and drug removal method capable of simultaneously removing leukocytes and an inactivated drug from a blood product containing red blood cells at a high rate in a short time while suppressing hemolysis, leukocytes and It is an object to provide a drug removal filter and a leukocyte and drug removal system.

  As a result of intensive research aimed at achieving the above-mentioned object, the present inventors appropriately used a leukocyte removal medium such as a non-woven fibrous structure capable of removing leukocytes and a drug removal medium such as fibrous activated carbon capable of removing the drug. Knowledge that it is possible to provide a flow-type removal method and a removal system that can remove leukocytes and inactivated drugs at the same time while suppressing hemolysis and at a high rate in a short time by placing them in a container with a simple arrangement Got.

  The leukocyte and drug removal method of the present invention is a single filter comprising an inactivated drug added to a blood product containing red blood cells, the inactivated blood product containing a leukocyte removal medium on the upstream side, and a drug removal medium on the downstream side. And removing it from the outlet to remove leukocytes on the upstream side of the filter and remove the remaining inactivated drug on the downstream side.

  The leukocyte and drug removal filter of the present invention is a filter that removes leukocytes and an inactivated drug from a blood product after inactivation of an inactivation target contained in a blood product containing red blood cells, and includes red blood cells. A container having an inlet through which blood products are introduced and an outlet through which blood products are discharged; a leukocyte-removing medium that is housed in the container and capable of removing white blood cells; A leukocyte removal medium, and the leukocyte removal medium is disposed upstream of the drug removal medium.

  According to these inventions, leukocytes are removed on the upstream side of the filter, and the remaining inactivating agent is removed on the downstream side, so that the leukocytes and the inactivating agent can be simultaneously removed at a high rate in a short time while suppressing hemolysis. Can be removed.

  The leukocyte removal medium may be a non-woven fiber structure, and the drug removal medium may be fibrous activated carbon. Activated carbon is used as the drug removal medium, and in particular, by making the activated carbon form fibrous rather than particles, the pore surface area per unit weight of the activated carbon is increased, and high drug removal ability is obtained. Moreover, since the space | gap which a blood cell passes is large, the increase in the pressure loss of a filter can be suppressed and the hemolysis of a blood formulation can be suppressed more effectively. In addition, fibrous activated carbon is suitable for mass production because it is excellent in moldability in production, quality control is relatively simple, and high chemical removal performance can be reproduced with the same design. In addition, when the non-woven fiber structure is manufactured from a thermoplastic material, the fibrous activated carbon is combined with the non-woven fiber structure and the container, and is integrally formed by applying energy such as heat and pressure, or bonded by an adhesive material. Can be formed integrally, which is advantageous in manufacturing.

Further, when the specific surface area of the fibrous activated carbon is 900 m ² / g or more and 3000 m ² / g or less, and the ventilation pressure loss of the leukocytes and the drug removal filter is 500 Pa or more and 2000 Pa or less, a higher removal rate can be obtained. At the same time, it is preferable because hemolysis can be suppressed more effectively.

  Here, the aeration pressure loss is an index representing the resistance value of the filter, and is an index suitable for simply selecting whether the degree of hemolysis and the leukocyte removal rate of the blood product after filtration achieves a predetermined purpose. The airflow pressure loss referred to in the present invention is determined by measuring the difference between the inlet pressure and the outlet pressure generated in the filter portion when a gas having a constant linear velocity is passed through the leukocyte and drug removal filter.

  The leukocyte and drug removal system of the present invention collects a blood product from which leukocytes and drugs have been removed by the leukocyte and drug removal filter, a blood product storage means for storing a blood product, and a leukocyte and drug removal filter. Blood product recovery means. According to this system, leukocytes are removed on the upstream side of the filter, and the remaining inactivated drug is removed on the downstream side, so that the leukocytes and the inactivated drug can be simultaneously reduced while suppressing hemolysis and at a high rate in a short time. The blood product from which leukocytes and inactivating drugs have been removed can be recovered.

  According to the present invention, leukocytes and inactivated drugs can be simultaneously removed from blood products containing red blood cells at a high rate in a short time while suppressing hemolysis.

It is sectional drawing of the leukocyte and chemical | medical agent removal filter which concerns on embodiment of this invention. It is a figure which shows the structure of the leukocyte and chemical | medical agent removal system which concerns on embodiment of this invention.

Hereinafter, preferred embodiments of a leukocyte and drug removal filter according to the present invention will be described in detail with reference to the drawings.
[Embodiment]
(Leukocyte and drug removal filter)

  First, the leukocyte and drug removal filter 1 according to the embodiment will be described with reference to FIG. The leukocyte and drug removal filter 1 is a column that simultaneously removes leukocytes and an inactivated drug from a blood product after the inactivation target in the blood product containing red blood cells is inactivated. In particular, the leukocyte and drug removal filter 1 according to the present embodiment is used alone as a single filter, and from a blood product containing red blood cells, leukocytes and an inactivated drug are simultaneously suppressed at a high rate in a short time while suppressing hemolysis. The purpose is to remove. The blood product as used herein refers to various blood products containing red blood cells, such as whole blood and red blood cell preparations. Further, the inactivation target in the blood product is, for example, leukocytes or pathogens, and the drug is, for example, a pathogen inactivation drug.

  Here, “pathogen inactivation” means depriving leukocytes or pathogens such as viruses, bacteria, protozoa and parasites from their ability to replicate or proliferate. The “pathogen inactivating drug” is a drug having a function of inactivating leukocytes mixed in blood products or pathogens such as viruses, bacteria, protozoa, and parasites. Moreover, the treatment for inactivation performed using an inactivating agent (white blood cell and pathogen inactivating treatment) is a treatment method in which a pathogen inactivating agent is added to a blood product and light is irradiated for a certain period of time as necessary. It is. An inactivating agent generates highly reactive compounds such as singlet oxygen, hydroxy radicals, and hydrogen peroxide by photosensitizing action, damages leukocytes and pathogen DNA or RNA, and DNA or RNA It inactivates leukocytes and pathogens by creating covalent bonds between bases and hindering leukocyte and pathogen self-replication behavior and viral protein expression. Examples of the pathogen inactivating agent include acridine derivatives, thiazine derivatives, phenothiazine derivatives, pyridine derivatives, porphyrin derivatives, coumarin derivatives, pyrimidine derivatives, riboflavin, psoralen derivatives, ethyleneimine multimers, and the like.

  The leukocyte and drug removal filter 1 includes a column (container) 2 having an inlet 2a and an outlet 2b, a non-woven fibrous structure 3b suitable for removing leukocytes contained in the column 2, and a fibrous shape suitable for drug removal. Activated carbon 4. The non-woven fiber structure 3b is an example of a leukocyte removal medium, and the fibrous activated carbon 4 is an example of a drug removal medium capable of adsorbing and removing a drug. The non-woven fiber structure 3 b is disposed upstream of the fibrous activated carbon 4. The inlet 2a of the column 2 is provided with an inlet connection 5a to which a blood circuit for introducing a blood product is connected. Similarly, the outlet 2b is provided with an outlet connecting portion 5b to which a blood circuit for discharging the blood product is connected.

  Further, in the present embodiment, a portion of the column 2 adjacent to the inlet 2a, that is, the uppermost stream with respect to the blood product flow, has a mean fiber diameter of 4 μm or more and 100 μm or less for the purpose of removing microaggregates. A woven fiber structure 3a is provided. Although a suitable aspect can be realized by providing the non-woven fiber structure 3a, the non-woven fiber structure 3a is not essential and can be omitted.

  The above-mentioned microaggregates are, for example, blood cell aggregates (aggregates), lipids, protein aggregates and the like containing fibrin and proteins generated during storage of blood products. By providing the non-woven fiber structure 3a, the microaggregates are completely removed before the blood product comes into contact with the non-woven fiber structure 3b. Therefore, good flowability is obtained in the subsequent leukocyte removal step and drug removal step. Can be secured. The material, thickness, and the like of the nonwoven fiber structure 3a can be determined as appropriate as long as the purpose of the leukocyte and drug removal filter 1 can be achieved, and there is no particular limitation.

  In the present embodiment, the mesh 3c is provided on the outlet 2b side of the column 2, that is, on the most downstream side with respect to the blood product flow. Although a suitable aspect can be realized by providing the mesh 3c, the mesh 3c is not essential and can be omitted.

  By providing the mesh 3c, when activated carbon fiber falls off during the blood product processing, it can be captured and mixed into the blood product can be avoided. It also has the effect of making the blood product being filtered more uniform and preventing single flow. The material, fiber diameter, pore diameter, thickness, and the like of the mesh 3c can be appropriately determined as long as the purpose of the leukocyte and drug removal filter 1 can be achieved, and is not particularly limited.

  A device that allows blood products to flow in like the leukocyte and drug removal filter 1 and contact the filter medium in a flow type is called an inline column. The method using an in-line column does not require a batch-type shaking device that brings a blood product and a filter medium into contact for a long time. Moreover, since the removal of leukocytes and drugs is completed in a short time, it is preferable because it is economical and efficient. The batch method is not particularly suitable for blood products containing red blood cells because the contact time between activated carbon and red blood cells tends to damage the red blood cell surface and cause hemolysis.

  The leukocyte and drug removal filter 1 first completes leukocyte removal by disposing the non-woven fiber structure (leukocyte removal medium) 3b upstream of the fibrous activated carbon (drug removal medium) 4 in the filter, Further, since the step of completing the removal of the drug is taken, clogging in the fibrous activated carbon 4 can be avoided, and good flowability of the blood product can be ensured.

Further, the specific surface area of the fibrous activated carbon 4 900m ² / g or more, is set to lower than or equal to 3000 m ² / g, sufficient drug removal capability can be obtained. Further, by setting the air pressure loss of the leukocyte and drug removal filter 1 to 500 Pa or more and 2000 Pa or less, a sufficient leukocyte removal rate can be obtained and physical friction between the red blood cells and the filter medium can be suppressed. It is advantageous.

  Here, the specific surface area is an index of the porosity of the porous body, and is an amount obtained by converting the surface area including the pore surface area of the porous body per unit weight of the porous body. The specific surface area can be obtained by measuring the pore distribution by a gas adsorption method or a mercury intrusion method based on the adsorption amount of gas molecules. The fibrous activated carbon 4 has a distribution peak particularly in micropores (8 to 20 cm) or mesopores (20 to 500 cm), and the pores on the surface of the activated carbon are particularly excellent in adsorption of low molecular weight drugs. Demonstrate.

  The airflow pressure loss is an index representing the resistance value of the filter, and is an index suitable for simply selecting whether the degree of hemolysis and the leukocyte removal rate of the blood product after filtration achieves a predetermined purpose. The airflow pressure loss referred to in the present embodiment is obtained by measuring the difference between the inlet pressure and the outlet pressure generated in the filter portion when a gas having a constant linear velocity is passed through the leukocyte and drug removal filter 1. A detailed method for measuring the airflow pressure loss is as follows.

  A gas whose aeration line speed is adjusted to 0.73 m / min by a membrane flow meter is introduced from the inlet 2a of the column 2 of the leukocyte and drug removal filter 1 and the air is freely discharged from the outlet 2b of the column 2. Thereafter, the difference between the inlet pressure and the outlet pressure when the flow rate was sufficiently stabilized was measured with a pressure transducer to determine the ventilation pressure loss (Pa).

The ventilation line speed here is the volume of the gas that passes through the unit cross-sectional area per unit time when a gas having a constant flow rate is caused to flow in the filter (for example, in the column 2). When the cross-sectional area of the filter is S (m ² ) and the flow velocity of the gas flowing from the filter container inlet side is v (L / min), the linear velocity of the air is expressed by v × 10 ⁻³ ÷ S (m / min). .
(Fibrous activated carbon)

The fibrous activated carbon 4 is housed in the column 2 and can be removed from the drug. In particular, any material that can reduce hemolysis of erythrocytes is sufficient. For example, mineral materials such as petroleum pitch, coal pitch, coal coke, tar peat, lignite, and resin materials such as phenol resin, cellulose resin, rayon, acrylic resin, vinylidene chloride resin, polyacrylonitrile fiber, polyvinyl alcohol fiber, etc. After a certain activated carbon raw material is melt-spun and molded, it is infusibilized, and after carbonization, it is activated by reacting with a gas containing water vapor at a high temperature. Alternatively, the fibrous activated carbon 4 can be formed by activating and molding the infusible and carbonized precursor fiber. The specific surface area is desirably 900 m ² / g or more and 3000 m ² / g or less. When the specific surface area is less than 900 m ² / g, the drug can be removed, but the removal rate may decrease.

  The average fiber diameter of the fibrous activated carbon 4 is preferably 10 μm or more and 100 μm or less. When the average fiber diameter is less than 10 μm, the mechanical strength is insufficient. Therefore, when blood cells pass in the filtration step, the fibrous activated carbon 4 is deformed by the pressure of the fluid, and the gap passing through the blood cells is too small. This tends to cause clogging of blood cells and an increase in pressure, and the effect of suppressing hemolysis tends to decrease. On the other hand, when the average fiber diameter is larger than 100 μm, the specific surface area required for drug removal cannot be obtained, and the drug removal rate tends to decrease. That is, when the average fiber diameter of the fibrous activated carbon 4 is 10 μm or more and 100 μm or less, there is an advantage in both the suppression of hemolysis and the drug removal rate.

  Although the shape of the fibrous activated carbon 4 of this embodiment is not specifically limited, Chop shape, felt shape, twisted yarn shape, woven fabric shape, paper shape, etc. are mentioned. In any case, if the fibrous activated carbon 4 having an appropriate specific surface area is uniformly packed in the column so as to have an appropriate airflow pressure loss, materials having other shapes can be used. From the viewpoint of the uniformity of the basis weight of the fibrous activated carbon 4, the uniformity of the voids, the mechanical strength for maintaining the dimensions, etc., a felt shape and a woven fabric shape are more preferable.

  As a method for producing felt-like activated carbon, spinning is performed from mineral, natural resin, or synthetic resin raw materials to form felt fibers, followed by infusibilization, carbonization, activation treatment, and crimping of the fibers. In the process, a method of felting before or after infusibilization is known. (For example, JP-A-3-130447). Also known is a method in which a natural resin having a fiber-forming ability or a fiber derived from a synthetic resin is infusibilized, carbonized and activated to produce an activated carbon fiber and then felted (for example, Japanese Patent No. 4153529). A general felt processing process is a fiber blending process, a thin wrapping process with a spinning machine, several layers of wraps are compressed while applying heat and steam, and water, acid, or a weak alkaline solution is included, A step of shrinking by applying heat, pressure, vibration, etc., followed by a step of making the thickness and hardness uniform by bringing the fibers into contact with each other by hair cutting and hot pressing. Felt-like activated carbon is difficult to fray, and is easy to mold and maintain its shape.

  As a method for producing a woven activated carbon, a method is known in which a fiber is spun from a mineral raw material or a resin raw material, and then processed into a woven cloth, which is carbonized and activated (for example, a polyacrylonitrile type). JP-A-62-133124, phenol-resin system, JP-A-60-35509, JP-A-4153529, and polyvinyl alcohol system, JP-A-53-114925, JP-A-59-187624. (Kaisho 61-47827, JP2003-64535). A general woven fabric processing step includes a step of producing a single yarn by spinning natural fibers or chemical fibers, a spinning step such as combined yarn, twisted yarn, and steaming, a woven fabric step of combining warp and weft. Since the woven activated carbon is a plain weave shape, the thickness and hardness are uniform, and the shape is easily maintained.

  As an optimal embodiment, the fibrous activated carbon 4 is particularly preferably coated with a hydrophilic material having high biocompatibility including a hydroxyethyl methacrylate polymer. Any hydrophilic material can be used without particular limitation as long as it does not affect blood. For example, poly (2-hydroxyethyl methacrylate) (PHEMA), poly (N-isopropylacrylamide), poly (N, N-dimethylacrylamide), poly (vinyl alcohol), poly (N-vinyl-2-pyrrolidone) (PVP) ) And the like. Among these, hydrophilic polymers made of synthetic polymers such as PHEMA and PVP, which are particularly versatile and can reduce production costs, are preferable.

  It is preferable to coat the surface of the fibrous activated carbon 4 with the above-mentioned hydrophilizing material because the hydrophobic adhesion force and physical friction between the erythrocytes and the fibrous activated carbon 4 are reduced and hemolysis can be suppressed. Moreover, since the wettability with respect to the blood formulation of the fibrous activated carbon 4 improves and favorable fluidity | liquidity can be ensured, a chemical | medical agent removal can be implemented in a short time and it is preferable. Moreover, since the single flow can be suppressed by improving the wettability, it is also preferable in that the chemical removing ability of the fibrous activated carbon 4 can be sufficiently exhibited.

More preferably, the surface of the fibrous activated carbon 4 is coated with the hydrophilizing material and then further subjected to wet heat sterilization or radiation sterilization. Thereby, bridge | crosslinking arises on the surface of the fibrous activated carbon 4, and it can suppress that dust generate | occur | produces from the surface of the fibrous activated carbon 4 in the production process of the leukocyte and chemical | medical agent removal filter 1. Further, when the leukocyte and drug removal filter 1 is used, it is possible to suppress the fallen fibers from being mixed into the blood product.
(Nonwoven fiber structure)

  Specific examples of the nonwoven fiber structure 3b according to this embodiment include melt blown nonwoven fabric, flash nonwoven fabric, spunbond nonwoven fabric, spunlace nonwoven fabric, wet nonwoven fabric, dry nonwoven fabric, and other nonwoven fabrics as well as paper, woven fabric, mesh, etc. Woven fabrics. Examples of fiber materials include polyamide, polyester, polyacrylonitrile, polyurethane, polyvinyl formal, polyvinyl acetal, polytrifluorochloroethylene, poly (meth) acrylate, polysulfone, polystyrene, polyethylene, polypropylene, cellulose, cellulose acetate, hemp , Cotton, silk, glass, carbon, etc., all suitable.

  Moreover, as an optimal embodiment, the average fiber diameter of the nonwoven fiber structure 3b capable of removing leukocytes is preferably 0.7 μm or more and 2.2 μm or less. When the average fiber diameter is less than 0.7 μm, when the blood cells pass, the non-woven fiber structure 3b is deformed by the pressure of the fluid, and the gap through which the blood cells pass becomes too small, resulting in clogging and pressure increase. There is a tendency to reduce hemolysis suppression effect. On the other hand, when the average fiber diameter is larger than 2.2 μm, the size of the voids through which blood cells pass becomes excessive, and the surface area in contact with the white blood cells decreases, so that the white blood cell removal ability tends to decrease. That is, when the average fiber diameter of the nonwoven fiber structure 3b is 0.7 μm or more and 2.2 μm or less, there is an advantage in both suppression of hemolysis and improvement of leukocyte removal ability.

  Moreover, as an optimal embodiment, it is preferable that the nonwoven fiber structure 3b is covered with a hydrophilizing material. This is preferable because the hydrophobic adhesion force and physical friction between the red blood cells and the nonwoven fiber structure 3b are reduced, and hemolysis can be suppressed. In addition, the wettability of the nonwoven fibrous structure 3b to the blood product is improved, and good flowability can be secured, which is preferable because leukocyte removal can be performed in a short time. Moreover, since the single flow can be suppressed by improving the wettability, it is also preferable in that the leukocyte removing ability of the non-woven fibrous structure 3b can be sufficiently exhibited.

The non-woven fiber structures 3a, 3b, the mesh 3c, and the fibrous activated carbon 4 are combined so as to have a predetermined size before being stored in the column 2 of the leukocyte and drug removal filter 1. May be preformed.
(column)

  In this embodiment, the material of the column 2 for accommodating the filter medium in the leukocyte and drug removal filter 1 may be either a hard resin or a flexible resin. In the case of a hard resin, the material is a phenol resin or an acrylic resin. Epoxy resin, formaldehyde resin, urea resin, silicon resin, ABS resin, nylon, polyurethane, polycarbonate, vinyl chloride, polyethylene, polypropylene, polyester, styrene-butadiene copolymer, and the like. In the case of a flexible resin, it is preferably formed from a sheet or cylindrical molded product made of a flexible synthetic resin, and the material is soft polyvinyl chloride, polyurethane, ethylene-vinyl acetate copolymer, polyethylene or polypropylene. Such as polyolefin, hydrogenated product of styrene-butadiene-styrene copolymer, thermoplastic elastomer such as styrene-isoprene-styrene copolymer or hydrogenated product thereof, and thermoplastic elastomer and polyolefin, ethylene-ethyl acrylate, etc. A suitable material is a mixture with a softening agent. Preferred are polycarbonate, polyethylene, polypropylene, soft vinyl chloride, polyurethane, ethylene-vinyl acetate copolymer, polyolefin, and thermoplastic elastomers based on these, more preferred are polycarbonate, soft vinyl chloride, and polyolefin.

  The column shape is not particularly limited as long as it has a blood product inlet 2a and outlet 2b, but a columnar, polygonal columnar rigid container or flexible container is suitable. In order to prevent close contact with the filter medium and secure the blood flow path, the inner surface of the column may be formed in an uneven shape. In particular, in the case of a flexible container, when the filtration pressure from the blood inlet 2a side and the blood after filtration are collected by a drop, the filter medium is pressed against the container on the blood outlet 2b side by the negative pressure, and the blood flow It is effective to make the inner surface of the blood outlet side uneven. As for the column molding method of the present embodiment, as long as the non-woven fiber structure 3b and the fibrous activated carbon 4 are sealed inside the filter and the shape can be maintained, a filter within the air pressure loss range intended by the present embodiment can be obtained. There is no particular limitation.

According to the leukocyte and drug removal filter 1 of the present embodiment having the above-described configuration, at least 85% or more of the drug can be removed even with a blood product having a relatively high concentration of leukocytes or an inactivated drug, and blood The number of remaining leukocytes per unit of preparation can be reduced to 1 × 10 ⁶ or less. Furthermore, hemolysis of the collected blood product can be suppressed.

In addition, the leukocyte and drug removal filter 1 is made of a material that cannot cause shape deformation that is a substantial problem, and maintains an appropriate shape and airflow pressure loss during actual use, and has sufficient leukocyte and drug removal performance. Can be maintained.
(Leukocyte and drug removal system)

  Next, the leukocyte and drug removal system 100 of the embodiment configured with the leukocyte and drug removal filter 1 according to the embodiment will be described with reference to FIG. FIG. 2 is a configuration diagram of the leukocyte and drug removal system 100.

  The leukocyte and drug removal system 100 includes a leukocyte and drug removal filter 1, a blood product bag (blood product storage means) 11 that stores a blood product, a drug supply unit 12 that supplies an inactivated drug, and an inactivation bag 13. And a treatment liquid recovery bag (blood product recovery means) 14 for recovering the blood product after completing the removal of the leukocytes and the drug, pipes L1 to L5, and clamps C1 to C4.

  Here, blood products are various blood products containing red blood cells, such as whole blood and concentrated red blood cell preparations. The drug supply unit 12 is connected to the inactivation back 13 by a pipe L1 such as a blood tube, and supplies the drug to the inactivation back 13 through the pipe L1. Blood product bag 11 is connected to pipe L2, and pipe L2 is SCD connected to pipe L1 at the start of processing. The inactivation bag 13 receives the blood product and the drug from the blood product bag 11 and the drug supply unit 12 via the pipes L1 and L2, and performs an inactivation process.

  The inactivation bag 13 and the inlet connection part 5a of the leukocyte / drug removal filter 1 are connected to each other by a pipe L3, and the treatment liquid recovery bag 14 and the outlet connection part 5b of the leukocyte / drug removal filter 1 are connected by a pipe L4. Are connected to each other. A blood product containing leukocytes and an inactivated drug is supplied to the leukocyte and drug removal filter 1 through the pipe L3. The leukocyte and drug removal filter 1 provided with the non-woven fiber structure 3b and the fibrous activated carbon 4 removes leukocytes and drug contained in the blood product. The blood product from which the leukocytes and the drug have been removed is collected in the treatment liquid collection bag 14 through the pipe L4.

Further, the pipes L3 and L4 branch to form a bypass path including a pipe L5 that bypasses the leukocyte and drug removal filter 1. The pipe L <b> 5 is used for venting the processing liquid recovery bag 14. The clamp C1 is provided closer to the inactivation back 13 than the place where the inactivation back 13 of the pipe L1 is connected to the SCD. The clamps C2, C3, and C4 are provided in the pipes L3, L5, and L4, respectively.
(Leukocyte and drug removal method)

Next, a leukocyte and drug removal method using the leukocyte and drug removal system 100 will be described.
(1) First, it is confirmed that all the clamps C1, C2, C3, and C4 are closed.
(2) The pipe L2 connected to the blood product bag 11 is SCD connected to the pipe L1.
(3) The clamp C1 is opened to open the pipe L1, and the drug is supplied from the drug supply unit 12 to the inactivation bag 13 through the pipe L1, and the blood product is not dropped from the blood product bag 11 through the pipes L2 and L1 due to a drop. Introduced into the bag 13 for activation.
(4) The clamp C1 is closed, the pipe L1 is sealed and cut from the clamp C1 on the inactivation back 13 side, and the blood product bag 11 and the pipe L1 on the drug supply unit 12 side are discarded.
(5) A drug and a blood product are mixed in the inactivation bag 13 and, if necessary, UV irradiation is performed for a predetermined time, and incubation (standing) is performed. Inactivation occurs during incubation.
(6) After completion of the incubation, the blood product is mixed again in the inactivation bag 13, the clamps C2 and C4 are opened, and the blood product is transferred from the inactivation bag 13 to the leukocyte and drug removal filter 1 through the pipe L3. The white blood cell and the drug removal filter 1 are filtered out. The blood product filtered by the leukocyte and drug removal filter 1 is collected in the treatment liquid collection bag 14 through the pipe L4 (white blood cell and drug removal step).
(7) Open the clamp L3 and open the pipe L5.
(8) The air in the treatment liquid recovery bag 14 is released to the inactivated bag 13 through the pipe L5.
(9) Close the clamps C2, C3, C4.
(10) Seal and take out the processing liquid recovery bag 14.

  By the above operation, an inactivated drug is added to the erythrocyte preparation, and the inactivated blood product is converted into a leukocyte removal medium on the upstream side and a leukocyte and drug removal filter (single filter) 1 containing the drug removal medium on the downstream side. A leukocyte and drug removal method comprising a step of removing leukocytes on the upstream side of the leukocyte and drug removal filter 1 and removing the remaining inactivated drug on the downstream side by being introduced into the inlet 2a and discharged from the outlet 2b. realizable.

  As described above, the present invention has been described with reference to each embodiment. However, the present invention is not limited to the above-described embodiment, that is, this system is only an example of system construction, and construction of bags and circuits. The method, arrangement, and use conditions, method, and procedure of the system are not limited.

EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited by a following example.
Example 1
(Creation of leukocyte and drug removal filter)

The average fiber diameter of 1.2 [mu] m, basis weight 40 g / ^{m 2} of polyethylene terephthalate (PET) nonwoven fabric (nonwoven fabric structure) 3b 10 sheets of an average fiber diameter of 14.5 [mu] m, felted activated carbon having a basis weight of 90 g / ^{m 2} (fibers Four activated carbons) 4 were laminated in this order. This laminated filter medium is cut into 72 × 72 mm ² squares, filled in the column 2 of the hard container, and welded using an ultrasonic welding method so that the effective filtration area is 67 × 67 mm ^2. Filter 1 was created.
(Creation of leukocyte and drug removal circuit)

The leukocyte and drug removal filter 1 is disposed between the inactivation bag 13 and the treatment liquid recovery bag 14, and the conduit connected to the inactivation bag 13 is connected to the inlet connection portion 5 a of the leukocyte and drug removal filter 1. Then, the conduit connected to the treatment liquid recovery bag 14 was connected to the outlet connection portion 5b of the leukocyte and drug removal filter 1, respectively. Moreover, as each conduit, a tube made of polyvinyl chloride having an inner diameter of 3.0 mm and an outer diameter of 4.2 mm was used.
(Specific surface area measurement of fibrous activated carbon)

A necessary amount of the above-mentioned fibrous activated carbon 4 is put in a measuring cell of an automatic specific surface area measuring apparatus Tristar 3000 manufactured by Shimadzu Corporation and sealed, and the sample degassing apparatus VacPrep061 manufactured by Shimadzu Corporation is used for 60 or less at 100 millitorr or less. It was 900 m ² / g as a result of performing degassing for a minute and measuring the specific surface area by the gas adsorption method at adsorption gas: nitrogen gas and adsorption temperature: liquid nitrogen temperature.
(Evaluation of air pressure loss value)

A gas whose aeration linear velocity was adjusted to 0.73 m / min by a membrane flow meter was introduced from the inlet 2a of the column 2 of the leukocyte and drug removal filter 1 and the gas was freely discharged from the outlet 2b of the column 2. Thereafter, after the flow rate was stabilized, the difference between the inlet pressure and the outlet pressure of the filter portion was measured with a pressure transducer. As a result, the ventilation pressure loss was 500 Pa.
(Drug filtering method by gravity drop)

The above-described leukocyte and drug removal method and drug filtration method using the system are as follows. A solution prepared by dissolving methylene blue in physiological saline to a concentration of 1 μM was adjusted to an amount suitable for the leukocyte and drug removal filter 1 and enclosed in an inactivation bag 13. Next, leukocyte drug filtration was carried out by flowing in from the inlet 2a of the column 2 of the leukocyte and drug removal filter 1 by gravity drop and discharging from the outlet 2b.
(Measurement method of drug removal rate)

The methylene blue solution before and after filtration was placed in a quartz cell and set in a UV-visible spectrophotometer V-570 manufactured by JASCO, and absorbance at 644 nm was measured. The concentration was converted with a calibration curve obtained from a methylene blue solution having a known concentration, and the drug removal rate was calculated by the following equation.
Drug removal rate (%) = (methylene blue concentration mM after filtration) / (methylene blue concentration mM before filtration) × 100
(Human blood filtration method using gravity drop)

The human blood filtration method using the leukocyte and drug removal method and system is as follows. Human whole blood collected from a healthy person was enclosed in an inactivation bag 13 in an amount suitable for the leukocyte and drug removal filter 1. Next, human whole blood sealed in the inactivation bag 13 was introduced by gravity drop from the inlet 2a of the column 2 of the leukocyte and drug removal filter 1 and discharged from the outlet 2b.
(Human whole blood collection method)

The human whole blood was obtained from a healthy person at a rate of adding 14 ml of CPD as an anticoagulant to 100 ml of whole blood. The collected whole blood was stored at room temperature of 22 ° C. and used within 4 hours after blood collection.
(Evaluation method of leukocyte removal ability)

The leukocyte removal rate was measured as an index of leukocyte removal ability. The concentration of leukocytes in human whole blood before and after filtration was measured using a LeucoCOUNT kit manufactured by Nippon Becton Dickinson and a flow cytometer FACSCanto II, and the leukocyte removal rate was calculated from the following equation.
Leukocyte removal rate = log ₁₀ {white blood cell concentration in human whole blood after filtration (/ ml) ÷ white blood cell concentration in human whole blood before filtration (/ ml)}
(Method for evaluating hemolysis)

As an indicator of hemolysis of blood products, supernatant hemoglobin (Hb) concentration was measured as follows. The filtered human whole blood was placed in a blood collection tube and centrifuged at 3000 rpm (1700 × g) for 15 minutes using a table top centrifuge 5100 manufactured by KUBOTA, and the supernatant was collected. Next, the sample was brought into contact with the tip of a micro cuvette manufactured by Hemokyu Corporation, and the supernatant was sucked up. This micro cuvette was set in a free / low-concentration hemoglobin measuring device manufactured by Hemokyu Corporation, and the supernatant Hb concentration was measured. Here, it is preferable that the supernatant Hb concentration does not exceed 0.02 g / dL.
(Evaluation method for flowability)

  As an index of human blood flowability with respect to the leukocyte and drug removal filter 1, the time required for the filtration treatment was measured as follows. The filtration start time is the time when the whole blood starts to move vertically downward from the inactivation bag 13 toward the inlet 2a of the column 2 of the leukocyte and drug removal filter 1 due to a drop, and is measured by a stopwatch S056 manufactured by SEIKO. Started. The end point of the filtration was the time when the blood filled between the container on the inlet 2a side and the filter medium disappeared, and the stopwatch measurement was finished.

From the evaluation results of Example 1, by setting the specific surface area of fibrous activated carbon to 900 m ² / g and the ventilation pressure loss of leukocytes and drug removal filter 1 to 500 Pa, the drug removal rate is 88% and the white blood cell removal rate is −3. 5 log ₁₀ , the treatment time was 11.2 minutes, and the supernatant Hb concentration was 0.00 g / dL.
(Example 2)

In the leukocyte and drug removal filter 1, 22 sheets of the same non-woven fabric 3b made of PET as in Example 1, 4 sheets of felt-like activated carbon 4 having the same average fiber diameter and basis weight as in Example 1, but different in the degree of micropores, totaled A white blood cell and drug removal filter 1 was prepared and evaluated in the same manner as in Example 1 except that 26 sheets were laminated in this order. The specific surface area of the fibrous activated carbon measured by the same method as in Example 1 was 1800 m ² / g, and the air pressure loss of the leukocyte and drug removal filter 1 was 1200 Pa.

From the evaluation results of Example 2, by setting the specific surface area of the fibrous activated carbon to 1800 m ² / g and the ventilation pressure loss of the leukocytes and the drug removal filter 1 to 1200 Pa, the drug removal rate is 99.5% or more, and the leukocyte removal rate is -4.2 log ₁₀ , treatment time was 16.1 minutes, and supernatant Hb concentration was 0.01 g / dL.
(Example 3)

In the leukocyte and drug removal filter 1, 32 sheets of the same non-woven fabric 3b made of PET as in Example 1, 4 sheets of felt-like activated carbon 4 having the same average fiber diameter and basis weight as in Example 1, but different in the degree of micropores, totaled A white blood cell and drug removal filter 1 was prepared and evaluated in the same manner as in Example 1 except that 36 sheets were laminated in this order. The specific surface area of the fibrous activated carbon measured by the same method as in Example 1 was 3000 m ² / g, and the air pressure loss of the leukocyte and drug removal filter 1 was 2000 Pa.

From the evaluation results of Example 3, by setting the specific surface area of the fibrous activated carbon to 3000 m ² / g and the ventilation pressure loss of the leukocytes and the drug removal filter 1 to 2000 Pa, the drug removal rate is 99.5% or more, and the leukocyte removal rate is -4.8 log ₁₀ , treatment time was 23.6 minutes, and supernatant Hb concentration was 0.01 g / dL.
Example 4

In the leukocyte and drug removal filter 1, ten sheets of the same non-woven fabric 3b made of PET as in Example 1, four sheets of felt-like activated carbon 4 having the same average fiber diameter and basis weight as in Example 1 and different degrees of micropores, totaling A white blood cell and drug removal filter 1 was prepared and evaluated in the same manner as in Example 1 except that 14 sheets were laminated in this order. The specific surface area of the fibrous activated carbon measured by the same method as in Example 1 was 750 m ² / g, and the air pressure loss of the leukocyte and drug removal filter 1 was 500 Pa.

From the evaluation results of Example 4, by setting the specific surface area of the fibrous activated carbon to 750 m ² / g and the ventilation pressure loss of the leukocytes and the drug removal filter 1 to 500 Pa, the drug removal rate is 85% and the leukocyte removal rate is −3. 5 log ₁₀ , treatment time was 12.0 minutes, and the supernatant Hb concentration was 0.00 g / dL.
(Example 5)

In the leukocyte and drug removal filter 1, eight sheets of the same non-woven fabric 3b made of PET as in Example 1, four sheets of felt-like activated carbon 4 having the same average fiber diameter and basis weight as in Example 1 and different degrees of micropores, totaling A white blood cell and drug removal filter 1 was prepared and evaluated in the same manner as in Example 1 except that 12 sheets were laminated in this order. The specific surface area of the fibrous activated carbon measured by the same method as in Example 1 was 900 m ² / g, and the air pressure loss of the leukocyte and drug removal filter 1 was 400 Pa.

From the evaluation results of Example 5, by setting the specific surface area of the fibrous activated carbon to 900 m ² / g and the ventilation pressure loss of the leukocytes and the drug removal filter 1 to 400 Pa, the drug removal rate is 87% and the leukocyte removal rate is −3. 3 log ₁₀ , treatment time 8.4 minutes, supernatant Hb concentration was 0.00 g / dL.
(Example 6)

In the leukocyte and drug removal filter 1, 36 sheets of the same non-woven fabric 3b made of PET as in Example 1, and 4 sheets of felt-like activated carbon 4 having the same average fiber diameter and basis weight as in Example 1 but different in the degree of micropores, total A white blood cell and drug removal filter 1 was prepared and evaluated in the same manner as in Example 1 except that 40 sheets were laminated in this order. The specific surface area of the fibrous activated carbon measured by the same method as in Example 1 was 3000 m ² / g, and the aeration pressure loss of the leukocyte and drug removal filter 1 was 2250 Pa.

From the evaluation results of Example 6, by setting the specific surface area of the fibrous activated carbon to 3000 m ² / g and the ventilation pressure loss of the leukocytes and the drug removal filter 1 to 2250 Pa, the drug removal rate is 99.5% or more, and the leukocyte removal rate is -5.0 log ₁₀ , treatment time 28.2 minutes, supernatant Hb concentration was 0.02 g / dL.
(Comparative Example 1)

In the leukocyte and drug removal filter 1, 4 felt-like activated carbons 4 having the same average fiber diameter and basis weight as in Example 1 and different micropore opening degrees, and 22 PET nonwoven fabrics 3b as in Example 1, total A white blood cell and drug removal filter 1 was prepared and evaluated in the same manner as in Example 1 except that 26 sheets were laminated in this order. The specific surface area of the fibrous activated carbon measured by the same method as in Example 1 was 1800 m ² / g, and the air pressure loss of the leukocyte and drug removal filter 1 was 1200 Pa.

From the evaluation results of Comparative Example 1, by setting the specific surface area of fibrous activated carbon to 1800 m ² / g and the ventilation pressure loss of leukocytes and drug removal filter 1 to 1200 Pa, the drug removal rate is 99.5% or more, and the leukocyte removal rate is -3.9 log ₁₀ and the supernatant Hb concentration was 0.13 g / dL. Even after the treatment time of 60 minutes passed, the filtration was not completed due to the stop flow, so the process was stopped halfway.

The evaluation results of leukocyte and drug removal rate and supernatant Hb are shown in Table 1.

DESCRIPTION OF SYMBOLS 1 ... White blood cell and chemical | medical agent removal filter, 2 ... Column (container), 2a ... Inlet, 2b ... Outlet, 3a, 3b ... Nonwoven fiber structure (leukocyte removal medium), 4 ... Fibrous activated carbon (drug removal medium), 11 ... blood product bag (blood product storage means), 12 ... drug supply unit, 13 ... inactivation bag, 14 ... treatment liquid recovery bag (blood product collection means), 100 ... leukocyte and drug removal system.

Claims (7)

  An inactivated drug is added to a blood product containing red blood cells, and the inactivated blood product is introduced into the inlet of a single filter containing a white blood cell removing medium on the upstream side and a drug removing medium on the downstream side and discharged from the outlet. A leukocyte and drug removal method comprising a step of removing leukocytes upstream of the filter and removing residual inactivated drug downstream.   The leukocyte and drug removal method according to claim 1, wherein the leukocyte removal medium is a non-woven fiber structure, and the drug removal medium is fibrous activated carbon. 3. The leukocyte and drug removal according to claim 2, wherein the fibrous activated carbon has a specific surface area of 900 m ² / g or more and 3000 m ² / g or less, and the airflow pressure loss of the filter is 500 Pa or more and 2000 Pa or less. Method. A filter that removes leukocytes and inactivating agents from a blood product after inactivating an inactivated target contained in a blood product containing red blood cells,
A container having an inlet for introducing a blood product containing red blood cells and an outlet for discharging the blood product;
A leukocyte removal medium housed in the container and capable of removing the leukocytes;
A medicine removal medium housed in the container and capable of adsorbing and removing the medicine,
The leukocyte and drug removal filter, wherein the leukocyte removal medium is disposed upstream of the drug removal medium.   The leukocyte and drug removal filter according to claim 4, wherein the leukocyte removal medium is a non-woven fiber structure, and the drug removal medium is fibrous activated carbon. The specific surface area of the fibrous activated carbon is 900 m ² / g or more and 3000 m ² / g or less, and the aeration pressure loss of the leukocytes and the drug removal filter is 500 Pa or more and 2000 Pa or less. Leukocyte and drug removal filter. A leukocyte and drug removal system,
The leukocyte and drug removal filter according to any one of claims 4 to 6,
Blood product storage means for storing the blood product;
A leukocyte and drug removal system comprising: a blood product recovery means for recovering a blood product from which the leukocytes and drug have been removed by the leukocyte and drug removal filter.

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