JP5036026B2 - Blood component separation device and method of use thereof - Google Patents

Description

The present invention relates to an apparatus and method for separating collected blood from a plasma component and a non-plasma component by centrifugal force, and filtering the plasma component by a particle trapping filter when obtaining the plasma component.
In particular, this apparatus and method is suitable when using a filter having a small pore size to remove minute pathogens such as viruses from plasma components. Plasma components with the pathogen removed can be transfused into the patient.

  Processes for obtaining plasma components from whole blood collected from blood donors can be roughly classified into two types: whole blood collection and component blood collection. In whole blood collection, the collected whole blood is collected as it is, and is divided into a plasma component and a non-plasma component by a centrifuge as necessary in a facility such as a blood center and collected. On the other hand, in component blood collection, the collected whole blood is separated into a plasma component and a non-plasma component by a blood collection device at the time of blood collection, and only the plasma component is collected and the non-plasma component is returned to the blood donor. For this reason, the physical burden on the blood donor is relatively small as compared to whole blood collection, and it is effective for the planned collection of blood according to the demand for each component.

  When separating whole blood into a plasma component and a non-plasma component in component blood collection, a centrifugal bowl of the type disclosed in Patent Document 1 called a Latham bowl is widely used, and disclosed in Patent Document 2. A system in which the housing is held in an annular shape is a typical separation device.

  Human or animal plasma or serum is used as various raw materials in plasma preparations, plasma fractionation preparations, biotechnology and the like, but there is a potential risk of virus contamination. So far, methods for removing these viruses have been proposed, and one of them is a virus removal method by membrane filtration. The advantage of this method over other methods is that it can be removed by its fractionation ability if the virus is larger than a specific size, regardless of the type of virus, the window period, and the presence or absence of the envelope. That is the point.

  As a method for obtaining a plasma component from whole blood collected from a blood donor and performing filtration using a filter, in Patent Document 3, a centrifugal bowl having a tubular core having a permeable side wall is rotated to rotate the plasma component and non-plasma. After separating the components, a method is disclosed in which the plasma components are permeated from the sidewalls into the core by the pressure of the non-plasma components.

  However, for the purpose of removing fine particles such as viruses, for example, a liquid feed mechanism capable of generating a large pressure is required to achieve a sufficient filtration rate by such a method. In addition, the circuit connecting the bowl to the blood collection needle and the blood bag needs to be strong enough to withstand such a large pressure, and has the disadvantage that the manufacturing cost of the circuit increases. For example, in the case of using a peristatic pump that is generally used widely as a liquid feeding mechanism, it is necessary to operate the pump at a high speed to generate a high permeation pressure, thereby damaging the circuit. There is a possibility. In addition, the permeable side wall is preferably hydrophilic on the surface of the side wall in order to prevent adsorption of proteins in the plasma component, but such a side wall is wetted by the plasma component and the pores are derived from the plasma component. When the liquid is filled with liquid, the liquid is strongly held in the pores by capillary action, so that a large pressure is required to introduce the gas from the bowl outlet and return the separated non-plasma component. In addition, it is conceivable that the return blood flow fluctuates in accordance with the discharge of the liquid from the pores, and there is a drawback that the control becomes difficult.

That is, a blood component separation device and a separation method are not known in which a plasma component and a non-plasma component are separated from whole blood by centrifugation and then minute pathogens such as viruses are rapidly removed from the plasma component by filter filtration. It was.
U.S. Pat. No. 4,300,177 Japanese Patent No. 2556741 US Pat. No. 6,464,624

  The present invention separates whole blood into a plasma component and a non-plasma component by centrifugal force, and at the same time, removes a minute pathogen such as a virus mixed in the obtained plasma component, and a method of using the device The purpose is to provide.

  In particular, regardless of the type of pathogen, the window period, and the presence or absence of an envelope, pathogens of a specific size or more are removed by filtration through a filter, and the collection of filtered plasma components and blood donors of non-plasma components An object of the present invention is to provide a device capable of promptly returning blood to the patient and a method of using the device.

  As a result of intensive studies to solve the above problems, the present inventor has found that the first space for separating blood by centrifugal force and the second space having the particle trapping filter have the same rotation axis. It has been found that a minute pathogen such as a virus mixed in a plasma component can be quickly removed by a blood component separation apparatus having a passage connecting the two spaces and arranged so as to have the present invention. It came to complete.

That is, the present invention
1. A first space (A) for separating whole blood collected from a blood donor into a plasma component and a non-plasma component by centrifugal force, and a particle capturing filter for filtering the separated plasma component by centrifugal force The second space (B) having the same rotation axis and having a passage (C) for transferring a plasma component from the first space to the second space. A blood component separation apparatus comprising a centrifugal separator.
2. The blood component separation device according to 1 above, wherein the second space is located at least on the outer side in the centrifugal radial direction of the first space.
3. The blood component separation device according to 1 or 2 above, wherein the second space comprises a hard housing.
4. The second space described above, wherein the second space is partitioned by a particle trapping filter into two compartments inside and outside the centrifugal radial direction, and the two compartments are ventilated by connecting means. Blood component separation device.
5. The blood according to 4 above, wherein the connecting portion of the connecting means is disposed above the inlet of the discharge path for discharging the plasma component filtered by the particle supplement filter from the second space. Component separation device.
6. The blood component separation device according to 4 above, wherein an inlet portion of the discharge path is located on the outer side in the centrifugal radial direction than the particle trapping filter.
7. The blood component separation device according to any one of 1 to 6 above, wherein the first space and / or the second space is constituted by a flexible housing.
8. The blood according to any one of 1 to 7 above, wherein the particles are at least one of viruses, bacteria, fungi, protozoa, parasites, pathogenic prions, leukocytes, and cell-derived particles. Component separation device.
9. A method for separating and collecting a plasma component from whole blood collected from a blood donor, the plasma component comprising:
(1) transfer whole blood to the first space in the centrifuge;
(2) In the first space, whole blood is separated into a plasma component and a non-plasma component by centrifugal force;
(3) transferring the plasma component from the first space to the second space provided with the particle trapping filter in the centrifuge using the passage C;
(4) A blood component separation method characterized in that in the second space, a plasma component is obtained by filtering with a particle capturing filter by centrifugal force.
About.

According to the apparatus and method of the present invention, since a plasma component separated from whole blood by centrifugal force is filtered by a particle trapping filter using the centrifugal force, a minute pathogen such as a virus mixed in the plasma component is removed. It can be suitably applied to the removal. Furthermore, since both the separation of plasma components and the removal of viruses mixed therein can be achieved by centrifugal force, the device having the specific structure of the present invention allows the separation of plasma components and the viruses mixed therein. Of course, it is possible to carry out the removal in a sequential manner as well as simultaneously, so that it can be applied more suitably.
That is, according to the present invention, it is possible to prepare plasma from which pathogens are removed aseptically and rapidly. For example, prepared plasma from which viruses and the like have been removed can be used as safe blood plasma for transfusion.

Hereinafter, an example of a preferred embodiment of the present invention will be specifically described with reference to the drawings. However, the present invention is not limited to the specific preferred embodiments described in the accompanying drawings.
FIG. 1 is a diagram showing an embodiment of the configuration of an apparatus for separating and collecting plasma components from whole blood in the present invention. A housing 12 having an inlet 1 and an outlet 10 and an introduction path 2 for connecting the inlet 1 and the housing 12 and a discharge path 9 for connecting the outlet 10 and the housing 12 are formed. The inside of the housing 12 converts whole blood into plasma components by centrifugal force. It consists of a blood component separation space 3 which is a first space for separating non-plasma components, and a second space 16 for filtering the plasma components with a particle capturing filter by centrifugal force. The second space 16 is divided into a filtration space 5 and a recovery space 7, and the blood component separation space 3 and the filtration space 5 are connected by a passage 14. A partition 15 between the filtration space 5 and the recovery space 7 is provided with a particle trapping filter 6 and a connecting means 11. The housing 12 includes a hermetic seal 13 between the housing 12 and the discharge path 9 so that the housing 12 can rotate around the introduction path 2 and the discharge path 9. In order to define the volume of the blood component separation space, the core portion 18 is provided, and a hermetic seal is provided between the introduction path 2 and the core portion 18. The above structure represents that the first space and the second space share the same rotation axis.

  Although the material of a housing is not specifically limited, For example, the composite material which consists of an organic material or an inorganic material, or an organic material and an inorganic material may be sufficient. Of these, organic materials, particularly organic polymer materials are preferable because of excellent processability such as injection molding and cutting. Examples of the organic polymer include polymethylpentene, polymethyl methacrylate, polyvinyl chloride, polycarbonate, polyurethane, polyacrylonitrile, polyvinyl alcohol, polyvinyl acetal, polyester, polyamide, polystyrene, polysulfone, cellulose, cellulose acetate, polyethylene, polypropylene, Polyvinyl fluoride, polyvinylidene fluoride, polytrifluorochlorovinyl, vinylidene fluoride-tetrafluoroethylene copolymer, acrylonitrile-styrene copolymer, acrylonitrile-butadiene copolymer, polychlorofluoroethylene, polyethersulfone, poly ( (Meth) acrylate, butadiene-acrylonitrile copolymer, polyether-polyamide block copolymer, ethylene - Although vinyl alcohol copolymer, and the like, a housing of the present invention is not limited thereto.

  The material of the housing may be either hard or soft. For example, a hard organic material may be used to mold into a bowl shape with an internal section as shown in FIG. When a soft organic material is used, for example, a unit comprising a flexible blood separation housing 23 and a filtration flexible housing 24 having a particle trapping filter 25 as shown in FIG. Can be installed and used.

  The material of the introduction path and the discharge path is not particularly limited, and may be, for example, an organic material or an inorganic material, or a composite material composed of an organic material and an inorganic material. Of these, inorganic materials, particularly metals, are preferred because of their strength and workability. Examples of the metal include aluminum, titanium, iron, copper, chromium, molybdenum, magnesium, and alloys thereof, but the introduction path and the discharge path of the present invention are not limited to these. Moreover, you may comprise combining the part which consists of an inorganic material represented by the metal, and the part which consists of organic materials.

  The blood component separation space and the filtration space inside the housing are connected by a passage, so that the plasma component separated from the whole blood in the blood component separation space can be quickly transferred to the filtration space. There is no particular limitation on the position, size, and number of passages connecting the blood component separation space and the filtration space, but it is more in the blood component separation space that it is located inside the blood component separation space in the centrifugal radial direction. This is preferable because blood can be separated.

  There are no particular restrictions on the position, size, and number of blood component separation spaces and filtration spaces inside the housing, but the fact that the filtration space is located outside the blood component separation space in the centrifugal radial direction means that the blood component separation space and the filtration space are separated. The plasma component that has reached the connecting passage can be quickly transferred to the filtration space by centrifugal force, which is preferable.

  There is no particular limitation on the size and number of the filtration space located on the primary side and the collection space located on the secondary side with respect to the particle trapping filter. However, since the filtration is performed by centrifugal force, the collection space is a centrifuge of the filtration space. It is preferable to be located on the radially outer side. It is preferable that the filter surface of the particle trapping filter that divides the filtration space and the recovery space be positioned so as to be perpendicular to the centrifugal radius because a centrifugal force can be uniformly applied to the entire surface of the particle trapping filter.

  The particle trapping filter described in the present invention is a structure made of a porous material, has a large number of fine pores communicating from one surface to the other surface, and selectively separates specific particles. It means a substrate having a pore size or surface affinity that can be removed.

  The porous body refers to a substrate having a large number of fine pores, and the material, thickness, shape, dimensions and the like are not limited. The material of the particle trapping filter made of a porous material may be an organic material, an inorganic material, or a composite material made of an organic material and an inorganic material. Among them, an organic material, particularly an organic polymer material is a preferable material because it has excellent processability such as cutting. Examples of the organic polymer include polycarbonate, polyurethane, polyacrylonitrile, polyvinyl alcohol, polyvinyl acetal, polyester, polyamide, polystyrene, polysulfone, cellulose, cellulose acetate, polyethylene, polypropylene, polyvinyl fluoride, polyvinylidene fluoride, and polytrifluorochloro. Vinyl, vinylidene fluoride-tetrafluoroethylene copolymer, polyethersulfone, poly (meth) acrylate, butadiene-acrylonitrile copolymer, polyether-polyamide block copolymer, ethylene-vinyl alcohol copolymer, etc., but are not limited thereto. It is not something. Also, a plurality of types of organic polymers can be mixed in order to obtain desirable properties.

  The shape of the particle trapping filter made of a porous material is not particularly limited as long as it has pores capable of trapping particles, but a flat plate shape is preferable from the viewpoint of easy formation of the housing. Examples thereof include meshes, films, sheets, membranes, plates, nonwoven fabrics, filter papers, sponges, woven fabrics, knitted fabrics, beads and the like. When capturing the particles by filtration, the size of the holes for capturing the particles can be easily controlled so that the operation can be easily performed, and considering the ease and cost of manufacturing the filter and housing, mesh, film, sheet, filter paper , Membranes, plates, nonwoven fabrics, sponges and beads are preferred, and meshes, films, sheets, membranes and nonwoven fabrics are more preferred.

  In order to facilitate the selective capture of particles, a plurality of types of porous bodies or a plurality of porous bodies may be used as a filter, and may be bonded by applying heat or pressure when overlapping. .

  In the present specification, the particle means a substance that can be trapped in the pores of the filter by size or affinity, and may be either an organic substance or an inorganic substance. The particles include, for example, particles that are harmful to health. When the particles are harmful to health, for example, the safety of blood for transfusion can be improved by capturing the particles with a particle capturing filter.

Health harmful particles mean particles that can be harmful to health by entering the human body, such as viruses, bacteria, fungi, protozoa, parasites, pathogenic prions, leukocytes and others. A cell-derived particle such as a cell disruption product.
When trapping particles by size, the pore diameter of the filter pores is preferably smaller than the particle diameter, but if the particles can be prevented from flowing out to the secondary side of the filter during the filtration process, the pores of the particle trapping filter There is no limitation on the size, and in view of selectively capturing the above-mentioned health-damaging particles, the average pore diameter is preferably 0.05 nm or more and 500 nm or less, more preferably 1 nm or more and 200 nm or less. Most preferably, it is 10 nm or more and 100 nm or less.

  The average pore diameter of the porous body in the present invention is a value measured with a mercury porosimeter. When measuring with a mercury porosimeter, the pore diameter (diameter) showing the maximum peak in the pore diameter distribution curve is the average pore diameter referred to in the present invention. Therefore, it represents a diameter that particles having a diameter larger than the average pore diameter of the porous body are difficult to enter, and it does not mean that particles having a diameter larger than this will never enter.

  A nonwoven fabric is a cloth-like material in which a collection of fibers or yarns are chemically, thermally, or mechanically bonded regardless of knitting. If the fibers are in contact with each other and maintain a certain shape by friction or entanglement, it can also be said to be mechanically coupled. A woven fabric means a fabric made by crossing warp and weft.

  When blood or blood-derived components are in contact with a particle trapping filter composed of the porous material, blood or blood-derived components such as hemolysis, platelet activation, complement activation, and protein adsorption are used for blood transfusion. In order to suppress undesirable phenomena, surface treatment with a hydrophilic compound may be performed to such an extent that the pores of the porous body are not blocked. A surface coating or the like can be used.

Examples of hydrophilic compounds used for surface treatment by grafting include acrylic acid, methacrylic acid, glycidyl methacrylate, hydroxypropyl acrylate, hydroxyethyl acrylate, hydroxyethyl methacrylate, methoxyethyl acrylate, methyl methacrylate, ethyl acrylate, ethylhexyl acrylate, phenoxyethyl acrylate, etc. Hydrophilic monomers such as acrylic acid or methacrylic acid and polyhydric alcohol esters, and mixtures of monomers, as well as polyethylene oxide, polyglycidol, polyvinylpyrrolidone, or copolymers thereof having a crosslinkable vinyl group or allyl group. Etc. are preferably used. The above-mentioned hydrophilic polymer having a crosslinkable vinyl group or allyl group in the main chain or side chain can be coated on the surface of the particle capturing filter and crosslinked with heat, radiation, a crosslinking agent, etc. A hydrophilic polymer such as a copolymer can be coated on the surface of the particle capturing filter. Moreover, after adding diacrylate type compounds, such as polyethyleneglycol diacrylate, to a hydrophilic compound, it can react, and the obtained reaction product can also be bridge | crosslinked.
A part of the partition wall that separates the filtration space and the recovery space has an opening, and the peripheral part of the particle trapping filter is bonded to the opening so as to be fixed to form a part of the partition wall. In order to prevent leakage of unfiltered plasma components from The bonding method is not particularly limited, and examples thereof include high-frequency welding, ultrasonic welding, thermal welding, and solvent welding because they can be firmly and uniformly bonded. Further, since a large pressure due to centrifugal force is applied to the particle trapping filter, a support member for the particle trapping filter may be provided on the outer side in the centrifugal radial direction of the particle trapping filter. Examples of the support member include mesh, nonwoven fabric, net, woven fabric, knitted fabric, and yarn. Further, the openings may be formed in a lattice shape, a slit shape, or a porous shape, and a part of the partition wall may function as a support member.

  Since the particle trapping filter that can remove minute pathogens such as viruses has a very small pore size, particularly when the pores of the particle removal filter are filled with a liquid, a large pressure is required to permeate the gas. For example, when the filtration space and the recovery space are connected only via the particle trapping filter, when introducing whole blood into the blood component separation space at the start of processing, the gas present in the blood separation space and the filtration space When the non-plasma components remaining in the blood separation space are returned from the introduction path after being processed, the particles are trapped. Since the gas returns from the recovery space to the filtration space via the filter, there is a concern that the time required for the start of treatment and blood return will increase, and the burden on the blood donor's mind and body will increase.

  Having a means for ventilating the filtration space and the recovery space allows the gas to quickly pass between the filtration space and the recovery space at the start of processing and blood return, thus greatly reducing the processing time. Is possible and preferable. Since troubles at the time of blood collection give a negative impression to the blood donor, it is required to prevent the blood donor from being generated as much as possible in the blood donation system that is realized by the spontaneous good intention of the blood donor. Providing a means to connect the filtration space and the collection space in an air-permeable manner allows blood to be returned even when the particle trapping filter is clogged by filtration and impervious to gas. It is preferable because troubles can be prevented.

  There are no particular restrictions on the means for ventilating the filtration space and the collection space, but it is a means for preventing the pre-filtration plasma component existing in the filtration space from entering the collection space without going through the particle trapping filter. Is required. On the other hand, the ability to reduce the virus concentration to 1 / 10,000 or less in plasma components for blood transfusion is used as an indicator of virus removal performance, thus preventing contamination even with droplets and mist generated by vibration during processing. It becomes important. As such means, for example, a ventilation filter can be used as the connection means. The fact that the filtration space and the collection space are connected by the ventilation filter allows gas to pass between the filtration space and the collection section, and the plasma component before filtration existing in the filtration space is, for example, being processed. It is preferable because it can be prevented from being mixed into the plasma component after filtration existing in the recovery space as droplets or mist generated due to vibrations.

  The ventilation filter described in the present invention is a structure made of a porous body, has a large number of fine pores communicating from one surface to the other surface, allows gas to pass therethrough, and allows liquid to pass through. It means a substrate having a pore size or surface affinity that can be blocked.

  The material, thickness, shape, dimensions, etc. of the ventilation filter are not limited. The material of the ventilation filter made of a porous material may be an organic material, an inorganic material, or a composite material made of an organic material and an inorganic material. Among them, an organic material, particularly an organic polymer material is a preferable material because it has excellent processability such as cutting. Examples of the organic polymer include polycarbonate, polyurethane, polyacrylonitrile, polyvinyl alcohol, polyvinyl acetal, polyester, polyamide, polystyrene, polysulfone, cellulose, cellulose acetate, polyethylene, polypropylene, polyvinyl fluoride, polyvinylidene fluoride, and polytrifluorochloro. Vinyl, vinylidene fluoride-tetrafluoroethylene copolymer, polyethersulfone, poly (meth) acrylate, butadiene-acrylonitrile copolymer, polyether-polyamide block copolymer, ethylene-vinyl alcohol copolymer, etc., but are not limited thereto. It is not something. A hydrophobic surface is preferred to prevent liquids. Examples of the organic polymer having such surface properties, excellent workability, and available at low cost include polyethylene, polypropylene, polyvinyl fluoride, and polyvinylidene fluoride. Also, a plurality of types of organic polymers can be mixed in order to obtain desirable properties.

  The shape of the ventilation filter made of a porous material may be any shape as long as it has pores that allow gas to pass therethrough and block liquid, but is preferably a flat plate from the viewpoint of easy formation of the housing. Examples thereof include meshes, films, sheets, membranes, plates, nonwoven fabrics, filter papers, sponges, woven fabrics, knitted fabrics, beads and the like. When capturing the particles by filtration, the size of the holes for capturing the particles can be easily controlled so that the operation can be easily performed, and considering the ease and cost of manufacturing the filter and housing, mesh, film, sheet, filter paper , Membranes, plates, nonwoven fabrics, sponges and beads are preferred, and meshes, films, sheets, membranes and nonwoven fabrics are more preferred.

  In order to facilitate gas permeation and liquid blocking, the porous body may be used as a plurality of types or a plurality of layers as a ventilation filter. Also good.

  The pore size of the ventilation filter made of a porous material is not limited. However, in consideration of preventing the permeation of liquid, the average pore diameter is preferably 1 nm or more and 1000 nm or less, more preferably 20 nm or more and 500 nm or less. Most preferably, it is 50 nm or more and 250 nm or less.

  It has an opening in a part of the partition that separates the filtration space and the recovery space, and the peripheral part of the ventilation filter is bonded to the opening so that it is fixed to form a part of the partition. This is preferred in order to prevent leakage of unfiltered plasma components into the collection space. The bonding method is not particularly limited, and examples thereof include high-frequency welding, ultrasonic welding, thermal welding, and solvent welding because they can be firmly and uniformly bonded. Moreover, you may have the support member of the ventilation filter inside or the outer side of the ventilation filter. Examples of the support member include mesh, nonwoven fabric, net, woven fabric, knitted fabric, and yarn. Further, the openings may be formed in a lattice shape, a slit shape, or a porous shape, and a part of the partition wall may function as a support member.

  As shown in FIG. 1, since it is desirable to quickly transfer the filtered plasma component to a storage container such as a blood bag and freeze it, a discharge route for transferring the filtered plasma component from the collection space to the storage container 9 is preferably provided. Although the shape and position of the discharge path are not particularly limited, the fact that the opening 8 in the collection space of the discharge path is located on the outer side in the centrifugal radial direction than the particle trapping filter is caused by centrifugal force on the outer wall in the centrifugal radial direction of the collection space. The moved plasma component after filtration can be quickly transferred to a storage container, which is preferable. The closer the opening is to the outer wall in the centrifugal radial direction of the recovery space, the smaller the amount of remaining liquid can be reduced. However, the opening should be kept away from contact due to shaking of the housing during rotation. Further, in the housing having a means for connecting the filtration space and the recovery space in a ventilation manner, when the plasma component filtered after the rotation is stopped is collected, as shown in FIG. Since the gas permeation is prevented by contacting the connecting means 11, the connecting means 11 is preferably located above the recovery port 22 of the discharge path.

  Whole blood collected from the blood donor is transferred from the inlet 1 through the introduction path 2 to the component separation space 3 partitioned by the outer wall 17. By rotating the bowl-shaped housing 12, the whole blood transferred to the component separation space 3 is separated into the plasma component 19 and the non-plasma component 20 in the component separation space 3 by centrifugal force as shown in FIG. By continuing the transfer of whole blood to the component separation space 3, the separated whole blood fills the component separation space 3, and the liquid level eventually reaches the dam part 4. Further, by continuing the transfer of whole blood, the plasma component 19 passes over the dam portion 4 and fills the filtration space 5 as shown in FIG. A part of the wall of the filtration space 5 is constituted by the particle trapping filter 6, and the plasma component 21 passes through the particle trapping filter 6 and enters the recovery space 7 by the centrifugal force acting on the plasma component 21 in the filtration space 5. enter. The plasma component that has entered the recovery space 7 is transferred from the recovery port 8 to the outlet 10 through the discharge path 9.

  FIG. 5 is an example of another embodiment of an apparatus for separating and collecting plasma components from whole blood in the present invention. Whole blood collected from the blood donor is transferred from the inlet 29 of the flexible housing 23 to the blood component separation space 26. As shown in FIG. 6, the whole blood transferred to the blood component separation space 26 is rotated by the centrifugal force by rotating the rotor 37 in the direction of the arrow 32 so that the centrifugal force acts. They are separated in the blood component separation space 26. By continuing the transfer of the whole blood to the blood component separation space 26, as shown in FIG. 7, the separated whole blood fills the blood component separation space 26, and the liquid level is eventually formed by the convex portions 38. Reach the dam part 39. Further, by continuing the transfer of the whole blood, the plasma component 33 crosses the dam portion 39, passes from the outlet 30 of the flexible housing 23 through the transfer path 36, and the inside is separated from the primary space 27 and the secondary space by the particle trapping filter. The flexible housing 24 divided into the side spaces 28 reaches the primary side space 27 from the inlet 31. Due to the centrifugal force acting on the plasma component 35 in the primary side space 27, the plasma component 35 passes through the particle trapping filter 25 and enters the secondary side space 28. Plasma components that have entered the secondary space are recovered from the outlet 32 of the flexible housing 24.

  In the method of the present invention, in the method of obtaining a plasma component from whole blood collected from a blood donor, whole blood is separated into a plasma component and a non-plasma component by centrifugal force in the first space of the blood component separation apparatus, The plasma component is transferred to the second space of the separation device, and the plasma component is filtered using a centrifugal force by a particle trapping filter provided in the second space.

  Means for separating whole blood into a plasma component and a non-plasma component by centrifugal force using a difference in specific gravity is known. Separation of blood components at the same time as blood collection is preferable because desired components can be quickly prepared, so that it is easy to respond to fluctuations in demand, but may be performed after collecting the collected whole blood.

  The means for transferring the whole blood to the first space in the centrifuge is not particularly limited. For example, a pump method such as a peristaltic pump or a syringe pump, or a blood bag containing whole blood is crushed. In addition to active transfer such as a method of pushing out to a circuit, transfer by a method using a head by placing a blood bag containing whole blood higher than a centrifuge is exemplified.

  The means for transferring the plasma component separated by centrifugal force in the first space to the second space provided with the particle trapping filter is not particularly limited. The method of extruding the plasma component to the second space by transferring whole blood continuously or intermittently to the first space without disturbing the interface between the plasma component and the non-plasma component that have already been separated is controlled by Since it becomes easy, it is preferable.

  The form when the plasma component is filtered by the particle trapping filter by centrifugal force is not particularly limited, but applying the centrifugal force in the vertical direction with respect to the filter surface can effectively utilize the filter surface of the particle trapping filter, preferable. In the present invention, filtering by centrifugal force is not limited to filtering only by centrifugal force, and the transmembrane pressure difference may be increased by a pump or the like in addition to centrifugal force.

  When the housing of the centrifuge is rigid, it is considered that the primary space of the particle trapping filter in the first space and the second space at the start of operation is filled with gas or liquid, Since it is not preferred that the plasma component is diluted with a liquid, it is preferred that at least a portion is filled with gas. By transferring whole blood to the first space, pressure is applied to the gas filling the first space. Since the primary space of the particle trapping filter in the first space and the second space are connected by a passage, the primary space of the particle trapping filter in the second space further in accordance with the transfer of whole blood. Pressure is applied to the gas that satisfies If the applied pressure is not released, the increased blood pressure in the primary space of the particle trapping filter in the first space and the second space prevents the whole blood from being transferred, while the pore size of the particle trapping filter is very high. If it is small, the speed at which the gas permeates the particle trapping filter is very slow. The pressure applied to the primary space of the particle trapping filter in the first space and the second space is pushed by the pressure by the ventilating connection means to the secondary space of the particle trapping filter in the second space. Transmission is preferable because the above problems can be solved. The gas pushed out to the secondary space of the particle trapping filter in the second space can be transferred out of the centrifuge through the discharge path from the secondary space of the particle trapping filter in the second space.

  When the centrifuge housing is rigid, it is considered that the secondary space of the particle trapping filter in the second space is filled with gas or liquid at the start of operation. Since dilution is not preferred, it is preferred that at least a portion is filled with gas. In the secondary space of the particle trapping filter in the second space, the plasma component is filtered by the particle trapping filter by the centrifugal force and reaches the secondary space of the particle trapping filter in the second space. Pressure is applied to the gas that satisfies If the applied pressure is not released, the pressure increased in the secondary space of the particle trapping filter in the second space reduces the transmembrane pressure difference of the particle trapping filter, preventing filtration, while the particle trapping filter. If the pore size of the particles is very small, the speed at which the gas permeates the particle trapping filter is very slow, and in particular the surface of the particle trapping filter is hydrophilic and a liquid such as a plasma component fills the pores. In some cases, the liquid filled in the pores is strongly held by capillary action, so that almost no gas can permeate. Extruding the gas through the ventilating connection means by the pressure applied to the secondary side space of the particle trapping filter in the second space to the primary side space of the particle trapping filter in the second space is Since such a problem can be solved, it is preferable. In particular, when the transfer is stopped after a certain amount of whole blood has been transferred, the primary side space of the particle trapping filter in the second space becomes negative pressure by filtering the plasma component, so that the secondary side The gas pushed out in the space permeates to the primary side through the ventilating connection means, thereby preventing a decrease in transmembrane pressure difference.

  In the case of collecting blood components by collecting whole blood from a blood donor and collecting plasma components, returning the non-plasma components remaining in the first space to the blood donor is the body of the blood donor This is preferable because it can reduce the burden on the user. Even when blood components are separated from the stored whole blood, returning the non-plasma components remaining in the first space to a container such as a blood bag can be used as a transfusion product such as an erythrocyte product. ,preferable. The means for transferring the non-plasma component at the time of returning blood is not particularly limited, but active transfer using, for example, a peristatic pump is preferable in order to complete the operation quickly. As a means for carrying out active transfer, the pressure may be a negative pressure on the inlet side of the centrifuge, or a positive pressure on the discharge side. It is preferable for the gas to move from the secondary side space of the particle trapping filter in the space to the primary side space through the ventilating connection means because the non-plasma component can be quickly transferred.

  When the housing of the centrifuge is rigid, the filtered plasma component is recovered from the discharge path, thereby creating a negative pressure in the secondary space of the particle trapping filter in the second space. If the pressure is not released, the secondary space of the particle trapping filter in the second space becomes negative pressure, thereby preventing the collection of plasma components, while the pore size of the particle trapping filter is very small, The speed at which the gas permeates through the particle trapping filter is very slow, and the pores are filled, especially when the surface of the particle trapping filter is hydrophilic and a liquid such as a plasma component fills the pores. Since the liquid is strongly held by capillary action, the gas hardly permeates. Permeation of gas from the primary side space of the particle trapping filter in the second space by the ventilating connection means to the secondary side space of the particle trapping filter in the second space that is negative pressure causes the above problem. Since it can eliminate, it is preferable.

Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to the following examples.

[Example 1]
A centrifuge having the configuration of FIG. 1 was prepared. The volume of the blood component separation space 3 is about 500 ml, the volume of the filtration space 5 is about 250 ml, and the volume of the recovery space 7 is about 250 ml. The outer wall 17 of the blood separation space is inclined about 15 degrees with respect to the rotation axis, and the distance from the rotation axis to the lower end and the distance from the rotation axis to the particle removal filter 6 are both 10 cm. As the particle removal filter 6, a surface of a polyethylene flat microporous membrane having a film thickness of 100 μm and an average pore diameter of 35 nm was made hydrophilic by graft polymerization of 2-hydroxypropyl acrylate. As the connection means 11 for ventilation, a polyethylene flat microporous membrane having an average pore diameter of 200 nm and a film thickness of 20 μm was used. Pressure gauges were attached to the inlet 1 and the outlet 10, and the pressure in each of the following steps was measured. When the pressure exceeded 0.05 MPa, the operation was stopped for safety.
Step 1 The outlet 2 is opened to the gas containing bag. About 450 ml of whole blood is introduced at 50 ml / min from the inlet 1 using the peristatic pump while rotating the centrifuge at 5000 rpm, and then the peristatic pump is stopped and centrifuged for about 20 minutes.
Step 2 While continuing the rotation, about 200 ml of whole blood is introduced from the inlet 1 at 50 ml / min to stop the rotation. By reversing the rotation direction of the peristatic pump, about 250 ml of non-plasma components in the blood separation space 3 are removed. Return blood.
Step 3 After completion of blood return, the centrifugal separator is rotated again at 5000 rpm, and after 15 minutes, the plasma component filtered by the peristatic pump is recovered from the outlet 11.
In step 1, the maximum was 0.01 MPa on the inlet side and the maximum was 0.002 MPa on the outlet side. In step 2, the maximum was 0.0001 MPa or less on both the inlet side and the outlet side. In step 3, the maximum was 0.0001 MPa or less on both the inlet side and the outlet side. Through a series of steps, approximately 200 ml of filtered plasma component could be prepared.

[Comparative Example 1]
An apparatus similar to Example 1 was prepared in which the connection means for ventilation was changed to a microporous film to make a polyethylene film having no pores, thereby preventing ventilation. In Step 1, the operation was canceled because the inlet side pressure exceeded 0.05 MPa when about 400 ml was introduced.

[Example 2]
An apparatus similar to that of Example 1 was prepared. 100 ml of a virus solution prepared so that the infectious titer of bovine viral diarrhea virus (BVDV) was 10 ⁸ at TCID ₅₀ was added to 900 ml of whole blood, and the same operation as in Example 1 was performed. In either step, the pressure did not exceed 0.05 MPa on both the inlet side and the outlet side. When the infectious titer of the filtered and collected plasma was measured, the measurement limit was 10 ³ or less, and the logarithmic removal rate (LRV) was 4 or more.

[Comparative Example 2]
An apparatus similar to Example 1 except that the particle trapping filter was removed was prepared. When the same operation as in Example 2 was performed, the pressure did not exceed 0.05 MPa on both the inlet side and the outlet side in any step. When the infectious titer of the collected plasma was measured, it was 10 ⁷ and the LRV was less than 0.1.

  The blood component separation device of the present invention is suitable as a medical device for preparing plasma from which pathogens such as viruses have been removed from whole blood, and can simultaneously separate blood components and remove pathogens.

It is the schematic diagram which showed the one aspect | mode of the structure of the blood component separation apparatus of this invention. It is the model which showed the one aspect | mode of isolation | separation of the blood component in the blood component separation apparatus of this invention. It is the schematic diagram which showed the one aspect | mode of the filtration of the plasma component in the blood component separation apparatus of this invention. It is the schematic diagram which showed the one aspect | mode of the filtration of the plasma component in the blood component separation apparatus of this invention. It is the schematic diagram which showed the one aspect | mode of the structure of the blood component separation apparatus of this invention. It is the schematic diagram which showed the one aspect | mode of isolation | separation of the blood component in the blood component separation apparatus of this invention. It is the schematic diagram which showed the one aspect | mode of the filtration of the plasma component in the blood component separation apparatus of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Inlet 2 Introductory path 3 Blood component separation space 4 Dam part 5 Filtration space 6 Particle capture filter 7 Collection space 8 Collection port 9 Outlet path 10 Outlet 11 Connection means 12 Housing 13 Sealing seal 14 Passage 15 Partition wall 16 Second space 17 Blood Outer wall 18 of component separation space Core 19 Plasma component 20 Non-plasma component 21 Plasma component 22 Recovery port 23 Flexible housing 24 for blood separation Flexible housing 25 for filtration Particle capture filter 26 Blood component separation space 27 Primary side space 28 Secondary side space 29 Inlet 30 Outlet 31 Inlet 32 Centrifugal radial direction 33 Plasma component 34 Non-plasma component 35 Plasma component 36 Transfer path 37 Rotor 38 Convex part 39 Dam part

Claims (9)

A first space (A) for separating whole blood collected from a blood donor into a plasma component and a non-plasma component by centrifugal force, and a particle trapping filter for filtering the separated plasma component by centrifugal force The second space (B) shares the same axis of rotation, and the second space is located at least radially outward of the first space, and the first space A blood component separation apparatus comprising a centrifuge characterized by having a passage (C) that opens near the rotation axis of the space and that transfers a plasma component from the first space to the second space. The blood component separation device according to claim 1, wherein the second space is formed of a hard housing. Space of the second is divided into two compartments centrifugal radially inner and centrifugal radially outwardly by a particle capture filter, characterized in that said two compartments are vented by connection means, according to claim 1 or 2 The blood component separation device according to 1. The blood component according to claim 3 , wherein the connecting portion of the connecting means is disposed above the inlet of the discharge path for discharging the plasma component filtered by the particle supplement filter from the second space. Separation device. The blood component separation device according to claim 4 , wherein an inlet portion of the discharge path is located on the outer side in the centrifugal radial direction than the particle trapping filter.

The blood component separation device according to any one of claims 1 to 5 , wherein the first space and / or the second space includes a flexible housing. The blood component according to any one of claims 1 to 6 , wherein the particles are at least one of viruses, bacteria, fungi, protozoa, parasites, pathogenic prions, leukocytes, and cell-derived particles. Separation device. The passage (C) is configured such that blood components separated by centrifugal force in the whole blood filling the first space (A) reach the second space (B) across the dam. The blood component separation device according to any one of claims 1 to 6. A method for separating and collecting plasma components from whole blood collected from a blood donor using the blood component separation device according to claim 7 , wherein the plasma components are
(1) transfer whole blood to the first space in the centrifuge;
(2) In the first space, whole blood is separated into a plasma component and a non-plasma component by centrifugal force;
(3) The plasma component beyond the dam portion is continuously transferred from the first space to the second space including the particle trapping filter in the centrifuge using the passage C,
(4) A blood component separation method characterized in that in the second space, a plasma component is obtained by filtering with a particle capturing filter by centrifugal force.