JP6516006B2 - Filtration device and filtration method - Google Patents

Description

  The present invention relates to a filtration device and a filtration method for filtering a biological substance in a liquid.

  In recent years, a cell trapping system for filtering a biological substance in a liquid has been disclosed (see, for example, Patent Document 1). The cell capture system disclosed in Patent Document 1 captures cells by passing a liquid containing cells as a biological substance from the upper side to the lower side in the vertical direction of the filter.

WO 2015/019889

  The cell capture system of Patent Document 1 has a problem that clogging occurs due to the deposition of the biological material on the filter, and the biological material can not be separated from the liquid.

  An object of the present invention is to solve the above-mentioned problems, and it is an object of the present invention to provide a filtration device and a filtration method capable of suppressing clogging with a biological substance and separating a biological substance from a liquid.

The filtration device of one embodiment of the present invention is
A container portion having an inlet for introducing a liquid containing a biological substance and an outlet for discharging the liquid;
A membrane portion provided between the inlet and the outlet of the container portion and having a plurality of through holes;
Equipped with
The film portion has a first major surface, and a second major surface located above the first major surface.
The biological substance is separated from the liquid by passing the liquid from the first main surface of the membrane part to the second main surface.

The filtration method of one embodiment of the present invention is
A method of filtering a biological substance from a liquid, comprising
A container unit including an inlet for introducing a liquid containing a biological material and an outlet for discharging the liquid, and a plurality of through holes provided between the inlet and the outlet of the container. Providing a filtration device comprising a membrane unit,
Separating the biological substance from the liquid by passing the liquid from the first main surface of the membrane part to the second main surface located above the first main surface;
including.

  According to the present invention, it is possible to provide a filtration device and a filtration method capable of suppressing clogging by biological material and separating biological material from liquid.

The schematic block diagram of the filtration apparatus of Embodiment 1 which concerns on this invention Schematic which shows a part of film part in Embodiment 1 which concerns on this invention A schematic view of a part of the film portion of FIG. 2 as viewed from the thickness direction Flow chart showing the filtration method of Embodiment 1 according to the present invention The figure which shows operation | movement of the filtration apparatus of Embodiment 1 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 1 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 1 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 1 which concerns on this invention. Diagram showing the operation of the filtration device of the reference example Diagram showing the operation of the filtration device of the reference example Diagram showing the operation of the filtration device of the reference example Diagram showing the operation of the filtration device of the reference example The figure which shows the measurement result of the biological material concentration contained in the liquid in the filtration apparatus of Embodiment 1 which concerns on this invention. The figure which shows the measurement result of the biological substance concentration contained in the liquid in the filter apparatus of a reference example The figure which shows the measurement result of the leak rate of the biological material in the filtration apparatus of Embodiment 1 which concerns on this invention. The figure which shows the measurement result of the leak rate of the biological material in the filtration apparatus of a reference example The schematic block diagram of the filtration apparatus of the modification of Embodiment 1 which concerns on this invention The schematic block diagram of the filtration apparatus of Embodiment 2 which concerns on this invention The figure which shows operation | movement of the filtration apparatus of Embodiment 2 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 2 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 2 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 2 which concerns on this invention. The schematic block diagram of the filtration apparatus of Embodiment 3 which concerns on this invention The figure which shows operation | movement of the filtration apparatus of Embodiment 3 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 3 which concerns on this invention. The schematic block diagram of the filtration apparatus of Embodiment 4 which concerns on this invention The figure which shows operation | movement of the filtration apparatus of Embodiment 4 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 4 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 4 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 4 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 4 which concerns on this invention. The figure which shows operation | movement of the filtration apparatus of Embodiment 4 which concerns on this invention.

The filtration device of one embodiment of the present invention is
A container portion having an inlet for introducing a liquid containing a biological substance and an outlet for discharging the liquid;
A membrane portion provided between the inlet and the outlet of the container portion and having a plurality of through holes;
Equipped with
The film portion has a first major surface, and a second major surface located above the first major surface.
The biological substance may be separated from the liquid by causing the liquid to pass from the first main surface to the second main surface of the membrane part.

  Such a configuration makes it possible to suppress clogging of the film portion due to the deposition of the biological material and to separate the biological material from the liquid.

  In the filtration device, the membrane portion may be a metal thin film.

  With such a configuration, deformation of the film portion can be suppressed even when a force is applied to the film portion.

  In the filtration device, at least a part of the membrane portion may be provided obliquely with respect to the vertical direction.

  Such a configuration can further suppress clogging of the membrane by the biological material.

  The filtration device may further include a suction unit that sucks the liquid from the inlet of the container unit to the outlet.

  With such a configuration, the biological substance can be separated from the liquid by sucking the liquid. In addition, biological substances can be separated from liquid in a short time.

The filtration device includes a liquid container for holding the liquid,
The inlet of the container portion is disposed in the liquid held in the liquid container,
The liquid container may be provided with a vent for releasing the pressure applied to the liquid.

  With such a configuration, the pressure applied to the liquid held in the liquid container can be released from the vent, so that the pressure applied to the biological substance in the liquid in the liquid container can be reduced, and damage to the biological substance can be reduced. Can be reduced. As a result, it is possible to suppress clogging due to deformation of the biological material in the membrane part.

  The filtration device may include a moving unit that moves the membrane unit downward.

  With such a configuration, the film portion can be maintained in the vicinity of the liquid surface by moving the film portion downward as the liquid decreases. Therefore, it is possible to further suppress clogging of the membrane portion with a biological substance.

The filtration method of one embodiment of the present invention is
A method of filtering a biological substance from a liquid, comprising
A container unit including an inlet for introducing a liquid containing a biological material and an outlet for discharging the liquid, and a plurality of through holes provided between the inlet and the outlet of the container. Providing a filtration device comprising a membrane unit,
Separating the biological substance from the liquid by passing the liquid from the first main surface of the membrane part to the second main surface located above the first main surface;
May be included.

  Such a configuration makes it possible to suppress clogging of the film portion due to the deposition of the biological material and to separate the biological material from the liquid.

  In the filtration method, in the separation step, the liquid may be allowed to pass through the membrane portion provided at least partially obliquely with respect to the vertical direction.

  Such a configuration can further suppress clogging of the membrane by the biological material.

  In the filtration method, the step of separating may include a step of suctioning the liquid from the inlet of the container portion to the outlet.

  With such a configuration, the biological substance can be separated from the liquid by aspirating the liquid. In addition, biological substances can be separated from liquid in a short time.

  In the filtration method, the separating may include releasing the pressure applied to the liquid.

  With such a configuration, the pressure applied to the liquid can be released. Therefore, in the membrane part, clogging due to deformation of the biological material can be suppressed. In addition, damage to biological material can be reduced.

  In the filtration method, the separating may include moving the membrane downward.

  With such a configuration, the film portion can be maintained in the vicinity of the liquid surface by moving the film portion downward as the liquid decreases. Therefore, it is possible to further suppress clogging of the membrane portion with a biological substance.

  Hereinafter, embodiments according to the present invention will be described with reference to the attached drawings. Moreover, in each figure, in order to make description easy, each element is shown exaggeratingly.

Embodiment 1
[overall structure]
FIG. 1 shows a schematic view of a filtration apparatus 100A according to a first embodiment of the present invention. As shown in FIG. 1, the filtration apparatus 100A includes a container unit 10 for introducing a liquid 60 containing a biological material 61, a membrane unit 20 for separating the biological material 61 from the liquid 60, and a liquid container for holding the liquid 60. 30, a suction unit 40 for suctioning the liquid 60, and a moving unit 50 for moving the membrane unit 20.

  In the present specification, the “biologically-derived substance” means a substance derived from an organism such as a cell (eukaryote), a bacterium (eubacteria), a virus and the like. Examples of cells (eukaryotic cells) include eggs, sperm, induced pluripotent stem cells (iPS cells), ES cells, stem cells, mesenchymal stem cells, mononuclear cells, single cells, cell clusters, suspension cells, adhesion Sex cells, nerve cells, white blood cells, lymphocytes, cells for regenerative medicine, autologous cells, cancer cells, circulating cancer cells in blood (CTC), HL-60, HELA, fungi. Examples of bacteria (eubacteria) include gram positive bacteria, gram negative bacteria, E. coli, and Mycobacterium tuberculosis. Examples of the virus include DNA virus, RNA virus, rotavirus, (bird) influenza virus, yellow fever virus, dengue fever virus, encephalitis virus, hemorrhagic fever virus and immunodeficiency virus. In the first embodiment, filtration apparatus 100A is particularly excellent in separating induced pluripotent stem cells (iPS cells), ES cells, stem cells, and circulating cancer cells in blood (CTC) from the liquid. In addition, the biological material may be a tissue in which cells are collected or a cell in which it is aggregated.

<Container section>
The container portion 10 has a tubular shape, and includes an inlet 11 for introducing the liquid 60 at one end and an outlet 12 for discharging the liquid 60 at the other end. As shown in FIG. 1, in the first embodiment, the container unit 10 is arranged such that the axial direction of the container unit 10 is the same as the vertical direction. The inlet 11 of the container unit 10 is positioned vertically below the outlet 12. A film unit 20 is provided at the inlet 11 of the container unit 10. Further, the inlet 11 of the container unit 10 is disposed in the liquid 60 held by the liquid container 30. The discharge port 12 of the container unit 10 is connected to the suction unit 40 via a tube 41. A moving unit 50 is attached to the outer wall of the container unit 10.

  The container portion 10 is formed of a resin such as acryl, epoxy, polystyrene, or polycarbonate, or a glass containing silicon oxide as a main component. The container portion 10 may be formed of a thermoplastic resin such as polypropylene, polystyrene or polycarbonate resin. It is preferable that the material of the container part 10 is a material which can be sterilized variously, such as gamma irradiation, an autoclave, ethylene oxide gas, and the like. The container portion 10 has, for example, an outer diameter of 1 mm or more and 60 mm or less, and an inner diameter of 0.5 mm or more and 50 mm or less and a height of 5 mm or more and 300 mm or less.

<Film section>
The membrane unit 20 is a porous membrane that separates the biological substance 61 from the liquid 60. Specifically, the film unit 20 is a metal thin film having a plurality of through holes 21. In the first embodiment, the film unit 20 is a circular metal mesh, has a pair of main surfaces facing each other, and is a structure having a plurality of through holes 21 penetrating both main surfaces.

  In the first embodiment, as shown in FIG. 1, the main surface on the lower side in the vertical direction is the first main surface VS1, and the main surface located on the upper side in the vertical direction than the first main surface VS1 is the second main surface VS2. Define as In the present specification, the upward direction in the vertical direction is the upper side, and the downward direction in the vertical direction is defined as the lower side. Further, in the film unit 20, the second main surface VS2 side is the upper side when viewed from the thickness direction of the film unit 20, and the first main surface VS1 side is defined as the lower side. The film unit 20 is provided at the inlet 11 of the container unit 10. In the first embodiment, the film unit 20 is arranged such that the first main surface VS1 and the second main surface VS2 are perpendicular to the inner wall of the container unit 10. That is, the first main surface VS1 and the second main surface VS2 of the film portion 20 are perpendicular to the direction (arrow 80 shown in FIG. 1) in which the liquid 60 flows from the inlet 11 to the outlet 12 of the container 10 It is arranged as.

  The plurality of through holes 21 are periodically arranged all over the first main surface VS1 and the second main surface VS2 of the film unit 20. The film unit 20 is formed of, for example, nickel. The dimensions of the film unit 20 are, for example, a diameter of 6 mm and a thickness of 1.2 μm. The material of the film unit 20 may be gold, silver, copper, platinum, iron, nickel, chromium, stainless steel, palladium, titanium, cobalt, and their oxides or alloys. In particular, gold, nickel, stainless steel, and titanium are preferable as the material of the film unit 20 when capturing the biological material 61. When the material of the film unit 20 is nickel, the surface of the nickel may be gold plated. Moreover, it is preferable that the material of the film | membrane part 20 is a material which can be sterilized variously, such as gamma ray irradiation, an autoclave, ethylene oxide gas.

  FIG. 2 shows a schematic configuration diagram of a part of the film unit 20 which is a two-dimensional periodic structure. The X, Y, and Z directions in FIG. 2 indicate the longitudinal direction, the lateral direction, and the thickness direction of the structure, respectively. FIG. 3 shows a part of the film unit 20 of FIG. 2 as viewed from the Z direction. As shown in FIG.2 and FIG.3, the film part 20 may be a plate-like structure (lattice-like structure) by which the several through-hole 21 is arrange | positioned at fixed intervals in matrix form. The film portion 20 is a plate-like structure in which a plurality of square through holes 21 are provided as viewed from the Z direction which is the main surface side. The plurality of through holes 21 are provided at equal intervals in two arrangement directions parallel to the sides of the square, that is, in the X direction and the Y direction in FIG. The through hole 21 is not limited to a square, and may be, for example, a rectangle, a circle, or an ellipse. Further, the arrangement of the holes is not limited to the square lattice arrangement. For example, in the case of a square arrangement, the spacing in the two arrangement directions may be an unequal rectangular arrangement, or a triangular lattice arrangement or a quasi-periodic arrangement. The through hole 21 may be provided with a taper.

  The shape and size of the through hole 21 of the membrane unit 20 are appropriately designed according to the size and shape of the biological substance 61 to be filtered.

<Liquid container>
The liquid container 30 is a container for holding the liquid 60 containing the biological substance 61. The liquid container 30 is a container having an opening larger than the diameter of the container portion 10. As shown in FIG. 1, the inlet 11 of the container unit 10 is disposed in the liquid 60 held in the liquid container 30. In the first embodiment, a gap is formed between the outer wall of the container unit 10 and the inner wall of the liquid container 30. The gap functions as a vent 31 for releasing the pressure applied to the liquid 60 held in the liquid container 30. In the first embodiment, since the liquid 60 is suctioned by the suction unit 40, pressure is applied to the biological substance 61 by negative pressure. In the filtering device 100A, since the liquid 60 can be opened to the outside by the vent 31, the pressure applied to the biological substance 61 in the liquid 60 can be released to the outside.

<Suction part>
The suction unit 40 sucks the liquid 60 held in the liquid container 30 from the inlet 11 of the container 10 to the outlet 12. The suction unit 40 is connected to the discharge port 12 of the container unit 10 via the tube 41. In the first embodiment, the suction unit 40 is a syringe. As shown in FIG. 1, the suction unit 40 suctions the liquid 60 held in the liquid container 30 by pulling the plunger of the syringe (arrow 81 shown in FIG. 1). Thereby, the liquid 60 in the liquid container 30 is sucked up from the inlet 11 of the container 10 to the outlet 12 by the suction unit 40 (arrow 80 shown in FIG. 1). That is, the liquid 60 passes from the first main surface VS1 of the film unit 20 to the second main surface VS2 located vertically above the first main surface VS1.

<Moving part>
The moving unit 50 moves the film unit 20 downward. Specifically, the moving unit 50 moves the film unit 20 toward the liquid 60 held in the liquid container 30. As shown in FIG. 1, in the first embodiment, the moving unit 50 is attached to the outer wall of the container unit 10. The moving unit 50 moves the container unit 10 vertically downward as the suction unit 40 reduces the liquid 60 in the liquid container 30 (arrow 82 shown in FIG. 1). Thus, the moving unit 50 can maintain the state in which the inlet 11 of the container unit 10 is disposed in the liquid 60 by moving the container unit 10 in the direction 82. In the first embodiment, the membrane unit 20 is disposed near the liquid surface of the liquid 60 held by the liquid container 30. The vicinity of the liquid level is a portion where the concentration of the biological substance 61 is low in the liquid container 30, and in Embodiment 1, the first main surface VS1 of the film portion 20 provided in the inlet 11 is at least liquid It is a position in contact with 60.

[Filtration method]
The filtration method according to the first embodiment will be described with reference to FIG.
FIG. 4 is a flowchart of the filtration method according to the first embodiment.

  As shown in FIG. 4, in step ST11, the filtration device 100A is prepared.

  In step ST12, the liquid 60 containing the biological material 61 is introduced into the liquid container 30.

  In step ST13, the inlet 11 of the container unit 10 is disposed in the liquid 60. Specifically, the inlet 11 of the container unit 10 is disposed in the liquid 60 held in the liquid container 30. The arrangement of the container unit 10 may be performed by the moving unit 50. The inlet 11 of the container unit 10 is disposed near the liquid surface of the liquid 60.

  In step ST14, the biological substance 61 is separated from the liquid 60 by passing the liquid 60 through the membrane unit 20. Step ST14 includes step ST15 of suctioning the liquid 60 in the liquid container 30 by the suction unit 40. In step ST15, the liquid 60 held in the liquid container 30 is sucked from the inlet 11 of the container unit 10 to the discharge opening 12 by pulling the plunger of the suction unit 40 (arrow 81 shown in FIG. 1) Arrow 80 shown in FIG. By suctioning the liquid 60 by the suction unit 40, the liquid 60 passes from the first main surface VS1 of the film unit 20 to the second main surface VS2 located vertically above the first main surface VS1. When the liquid 60 passes through the membrane unit 20, the biological substance 61 can not pass through the through hole 21 and remains in the liquid container 30.

  Step ST14 includes the step ST16 of releasing the pressure applied to the liquid 60 held in the liquid container 30. When the suction pressure is applied to the liquid 60 in the liquid container 30 by the suction unit 40, the biological substance 61 may be deformed. Therefore, the pressure applied to the biological substance 61 in the liquid 60 can be reduced by releasing the pressure applied to the liquid 60 from the vent 31 of the liquid container 30.

  Step ST14 may include step ST17 of moving the film unit 20 downward. By aspirating the liquid 60 by the suction unit 40, the liquid 60 held in the liquid container 30 decreases. By moving the container unit 10 downward toward the liquid 60 in the liquid container 30 by the moving unit 50, the inlet 11 of the container unit 10 can be maintained in the vicinity of the liquid surface of the liquid 60.

  As described above, in the filtration method according to the first embodiment, the biological substance 61 is separated from the liquid 60 by steps ST11 to ST16. Further, in the filtration method according to Embodiment 1, the liquid 60 which has passed through the membrane unit 20 can also be recovered.

[Operation of filtration device]
The operation of the filtration device 100A will be described in detail using FIGS. 5A to 5D.
5A-5D illustrate the operation of filtration device 100A. 5A to 5D, the suction unit 40 and the moving unit 50 are omitted to simplify the description.

  As shown in FIG. 5A, the inlet 11 of the container unit 10 is placed in the liquid 60 held in the liquid container 30 before the filtration is started. At this time, the film unit 20 is disposed near the liquid surface of the liquid 60.

  As shown in FIG. 5B, the suction unit 40 sucks the liquid 60 from the inlet 11 to the outlet 12 of the container unit 10 (arrow 80 shown in FIG. 5B). The liquid 60 in the liquid container 30 passes from the first main surface VS1 of the membrane unit 20 to the second main surface VS2. When the liquid 60 passes through the membrane unit 20, the biological substance 61 can not pass through the through holes 21 and is captured by the membrane unit 20. Therefore, the biologically-derived substance 61 starts to gather on the first main surface VS1 of the film unit 20.

  As shown in FIG. 5C, when the liquid 60 is continuously sucked in the direction 80, a cake layer A1 is formed on the first major surface VS1 of the film unit 20 by the biological material 61 being further collected. The cake layer A1 becomes thicker because the biological substance 61 further gathers in the membrane part 20 as the filtration time increases.

  As shown in FIG. 5D, when the thickness of the cake layer A1 is increased, the biologically-derived substance 61 repeats aggregation and detachment in the lower layer A2 of the cake layer A1. The aggregation and detachment of the biological substance 61 stop the increase of the thickness of the cake layer A1 and prevent clogging, so that the filtration does not stop. That is, in the filter device 100A, the cake layer A1 does not become thick enough to stop the filtration, so that the separation of the biological substance 61 can be continued.

The assembly and detachment of the biological substance 61 will be described in detail.
When the cake layer A1 is formed on the first main surface VS1 of the film unit 20, the through hole 21 of the film unit 20 is blocked by the cake layer A1, so that pressure resistance is generated. In the first embodiment, since the liquid 60 is suctioned by the suction unit 40, if an increase in pressure resistance occurs in the lower layer A2 of the cake layer A1, the suction force by the suction unit 40 is reduced. In addition, the biological material 61 in the liquid 60 is subjected to a force in the direction of gravity due to its own weight. In the lower layer A2 of the cake layer A1, when the suction force by the suction unit 40 is larger than the force in the direction of gravity, the biological substance 61 gathers in the cake layer A1. On the other hand, when the suction force by the suction unit 40 is smaller than the force in the direction of gravity, the biological substance 61 is separated from the cake layer A1. As described above, in the filtration device 100A, aggregation and detachment of the biological substance 61 are repeated by the balance between the suction force of the suction unit 40 and the force in the direction of gravity. The detachment of the biological substance 61 from the cake layer A1 can also be performed by vibration of the filtration device 100A. For example, the biological substance 61 may be separated from the cake layer A1 by vibration generated by shaking the filter device 100A up and down and / or left and right.

  Next, for comparison, the filtration device of the reference example will be described using FIGS. 6A to 6D.

  The filtration apparatus of the reference example includes a container portion 110 having an inlet 111 for introducing the liquid 60 and an outlet 112 for discharging the liquid 60, and a membrane portion 120 having a plurality of through holes. In the filtration device of the reference example, the inlet 111 is positioned above the outlet 112 in the vertical direction, and the membrane part 120 is provided at the outlet 112 of the container 110. In the filtration device of the reference example, the liquid 60 is allowed to pass from the second main surface VS12 of the membrane section 120 to the first main surface VS11 located vertically below the second main surface VS12. That is, while the filtration device 100A of the first embodiment passes the liquid 60 to the membrane portion 20 from the lower side to the upper side in the vertical direction, the filtration device of the reference example goes from the upper side to the lower side in the vertical direction. The liquid 60 is allowed to pass through the membrane unit 120. In the filtration apparatus of the reference example, the liquid 60 is allowed to pass through the membrane unit 120 by suctioning the liquid 60 by a suction pump installed below.

  As shown in FIG. 6A, in the filtration device of the reference example, the liquid 60 containing the biological substance 61 flows downward. That is, in the filtration device of the reference example, the liquid 60 flows from the inlet 111 of the container portion 110 toward the outlet 112 (arrow 80r shown in FIG. 6A). Therefore, the liquid 60 flowing in the container portion 110 passes from the second main surface VS12 of the film portion 120 to the first main surface VS11 located below the second main surface VS12 in the vertical direction. When the liquid 60 passes through the membrane 120, the biological substance 61 is captured by the membrane 120. For this reason, the biological material 61 is deposited on the second main surface VS12 of the film portion 120.

  As shown in FIG. 6B, when the liquid 60 continues to flow in the direction 80r, a cake layer A3 is formed on the second main surface VS12 of the film portion 120 by the deposition of the biological material 61.

  As shown in FIG. 6C, as the capture of the biological substance 61 by the membrane part 120 increases, the thickness of the cake layer A3 gradually increases. At this time, the filtration gradually shifts from the second main surface VS12 of the membrane part 120 to the upper layer A4 of the cake layer A3. That is, the separation of the biological substance 61 is not performed in the membrane part 120 but in the upper layer A4 of the cake layer A3.

  As shown in FIG. 6D, when the biological substance 61 is further deposited on the second major surface VS12 of the film portion 120, the thickness of the cake layer A3 further increases. Further, in the upper layer A4 of the cake layer A3, clogging due to the deposition of the biological material 61 progresses. When clogging increases in the upper layer A4 of the cake layer A3, the liquid 60 can not pass through the upper layer A4 of the cake layer A3 soon.

  Thus, in the filtration device of the reference example, the biological material 61 captured by the membrane unit 120 is deposited on the second major surface VS12 of the membrane unit 120. For this reason, since the cake layer A3 gradually increases with an increase in the amount of capture of the biological material 61 in the membrane part 120, clogging occurs in the upper layer A4 of the cake layer A3. As a result, in the filtration device of the reference example, the filtration is stopped by the clogging due to the deposition of the biological material 61 captured by the membrane unit 120, and the biological material 61 can not be separated from the liquid 60.

  On the other hand, in the filtration device 100A of the first embodiment, aggregation and detachment of the biological substance 61 are repeatedly performed in the lower layer A2 of the cake layer A1 formed on the first main surface VS1 of the membrane unit 20. For this reason, in the filtration device 100A, the filtration does not stop due to the clogging with the biological material 61. Therefore, the filtration device 100A can continue to separate the biological substance 61 from the liquid 60 by suppressing clogging compared to the filtration device of the reference example.

[effect]
<Filtration of high concentration liquid>
The filtration of the liquid 60 in the filtration device 100A and the filtration of the liquid 60 in the filtration device of the reference example will be described with reference to FIGS. 7 and 8.
FIG. 7 shows the measurement results of the concentration of the biological substance 61 when the liquid 60 is filtered by the filtration device 100A. In FIG. 7, the vertical axis indicates the time (min) required for filtration, and the horizontal axis indicates the concentration (cell / ml) of the biological substance 61. FIG. 8 shows the measurement results of the concentration of the biological substance 61 when the liquid 60 is filtered by the filter device of the reference example. In FIG. 8, the vertical axis indicates the time (min) required for filtration, and the horizontal axis indicates the concentration (cell / ml) of the biological substance 61. The concentration of the biological substance 61 means the number of the biological substance 61 per 1 ml. In addition, HL-60 which is a nucleated cell was used as a biological origin substance, and physiological saline was used as a solution.

The measurement conditions of filtration in FIGS. 7 and 8 will be described.
In the filtration apparatus of the filtration apparatus 100A and the reference example, after the liquid 60 was filtered and 80% of the liquid volume was extracted and discarded, the concentration of the liquid 60 containing the biological material 61 captured by the membrane part was measured respectively . In addition, the time required for the filtration of the liquid 60 was also measured.

Detailed conditions are shown below.
Membrane diameter: φ 6 mm
Through hole diameter: 2.5 μm
Hole spacing of through holes: 3.6 μm
Syringe suction speed at the start of suction: 0.2 ml / min
Volume: 1.0 ml
Liquid extraction volume: 0.8 ml

As shown in FIG. 7, in the filtration device 100A, when the liquid 60 is filtered and 80% of the liquid volume is extracted and discarded, the upper limit concentration of the biological substance 61 in the liquid 60 is about 1.0 × It was 10 ⁷ cells / ml. In addition, the time for filtering the liquid 60 to extract an 80% liquid amount was about 13 minutes.

As shown in FIG. 8, when the liquid 60 is filtered and 80% of the liquid volume is extracted and discarded in the filtration device of the reference example, the upper limit concentration of the biological substance 61 in the liquid 60 is about 2.0. It was 10 ⁶ cells / ml. In addition, it took about 20 minutes to filter the liquid 60 and extract an 80% liquid volume. In the filtration apparatus of the reference example, filtration of the liquid 60 having a concentration of the biological substance 61 of about 1.0 × 10 ⁷ cells / ml was tried, but the filtration did not proceed even after 1 hour.

  Thus, the filtration device 100A can filter the high concentration liquid 60 in a shorter time than the filtration device of the reference example. That is, compared with the filtration device of the reference example, the filtration device 100A has a higher limit concentration of the filtration of the biological substance 61, and can perform the filtration of the liquid 60 in a shorter time.

<Leakage rate of biological material>
The leak rate of the biological material 61 in the filtration device 100A and the leak rate of the biological material 61 in the filtration device of the reference example will be described with reference to FIG. 9 and FIG.
FIG. 9 shows the measurement results of the leak rate of the biological material 61 when the liquid 60 is filtered by the filtration device 100A. In FIG. 9, the vertical axis represents the leak rate (%) of the biological substance 61, and the horizontal axis represents the concentration (cell / ml) of the biological substance 61. FIG. 10 shows the measurement results of the leak rate of the biological material 61 when the liquid 60 is filtered by the filter device of the reference example. The measurement conditions of the leak rate of the biological material in FIGS. 9 and 10 are the same as the measurement conditions of the filtration in FIGS. 7 and 8 described above. The leakage rate means the percentage of biological material that has passed through the membrane. The leak rate is calculated by (number of biological substances passed through the membrane part) / (number of biological substances contained in the liquid before filtration) × 100.

  As shown in FIG. 9, in the filtration device 100A, when the liquid 60 is filtered and 80% of the liquid volume is extracted and discarded, the upper limit of the leak rate of the biological substance 61 is about 7%. That is, the recovery rate of the biological substance 61 in the filtration device 100A was about 93%.

  As shown in FIG. 10, in the filtration apparatus of the reference example, when the liquid 60 is filtered and 80% of the liquid volume is extracted and discarded, the upper limit of the leak rate of the biological substance 61 is about 15%. The That is, the recovery rate of the biological substance 61 in the filtration device 100A was about 85%.

  Thus, the filtration device 100A can achieve a higher recovery rate than the filtration device of the reference example. More specifically, in the filtration device of the reference example, the pressure in the vicinity of the membrane portion 120 may increase, and a biological material that originally has high deformability may be deformed and pass through the through hole of the membrane portion 120. . On the other hand, in the case of the filtration apparatus 100A, the pressure in the vicinity of the membrane unit 20 is relatively low, and the filtration operation can be performed while keeping the deformation of the biological substance small, Few. Therefore, the filtration device 100A can recover the biological substance 61 at a high recovery rate as compared to the filtration device of the reference example.

  According to the filtration device 100A in the first embodiment, the following effects can be obtained.

  In the filtration device 100A, the membrane unit 20 passes the liquid 60 containing the biological material 61 from the first main surface VS1 to the second main surface VS2 located vertically above the first main surface VS1. That is, in the filtration device 100, the liquid 60 is allowed to pass through the membrane unit 20 from the lower side to the upper side in the vertical direction to perform filtration. According to such a configuration, the biological material 61 can be separated from the liquid 60 while suppressing clogging of the film portion 20 due to the deposition of the biological material 61. As a result, in the filtering apparatus 100A, the higher concentration liquid 60 is filtered in a shorter time as compared with the filtering apparatus of the reference example in which the liquid 60 is allowed to pass through the membrane part 120 from the upper side to the lower side in the vertical direction. Can. In addition, in the filtration device 100A, the recovery rate of the biological substance 61 can be increased compared to the filtration device of the reference example. In Embodiment 1, filtration device 100A separates biological material 61 and liquid 60, but the present invention is not limited to this. For example, the filtering device 100A may filter foreign substances larger in size than the biological substance 61 contained in the liquid 60 by passing the solution in the liquid 60 and the biological substance 61. Alternatively, the filtering device 100A may filter and classify inorganic substances or organic substances of different sizes contained in the solution.

  In the filtration device 100A, the membrane unit 20 uses a metal thin film as a porous membrane. With such a configuration, damage to the film unit 20 by applying a force to the film unit 20 can be suppressed. In addition, even when the liquid 60 passes through the membrane unit 20, the through hole 21 of the membrane unit 20 is not easily deformed, so that the biological substance 61 is prevented from passing through the membrane unit 20 due to the deformation of the through hole 21 can do. Furthermore, damage to the membrane unit 20 by the liquid 60 can also be suppressed.

  In the filtration device 100A, a vent 31 for releasing pressure applied to the liquid 60 is provided by forming a gap between the outer wall of the container portion 10 and the inner wall of the liquid container 30. With such a configuration, the liquid 60 in the liquid container 30 can be open to the outside. Therefore, the pressure applied to the liquid 60 by suction can be released from the vent 31, and the pressure applied to the biological substance 61 in the liquid 60 can be reduced. As a result, deformation of the biological substance 61 can be suppressed, and clogging in the film unit 20 can be reduced. In addition, it is preferable that the gas which passes the vent 31 is sterilized. This is to prevent mixing of biological substances other than the biological substance 61 originally contained in the liquid 60. In addition, it may be connected via a vent 31 to a gas bag preferred by a biological material containing a suitable amount of carbon dioxide. Furthermore, when the volume of the gas bag is large, the pressure applied to the liquid 60 can be easily released by suction.

  In the filtration device 100A, the suction unit 40 sucks the liquid 60 held in the liquid container 30, thereby causing the liquid 60 to pass from the first main surface VS1 to the second main surface VS2 of the membrane unit 20. With such a configuration, the liquid 60 in the liquid container 30 can be allowed to pass through the membrane unit 20, and the biological substance 61 can be separated from the liquid 60. In addition, the suction unit 40 can perform filtration in a shorter time by suctioning the liquid 60.

  In the filter device 100A, the moving unit 50 moves the container unit 10 toward the liquid 60 as the liquid 60 in the liquid container 30 decreases. With such a configuration, it is possible to perform filtration while maintaining the state in which the membrane unit 20 is disposed near the liquid surface where the concentration of the biological substance 61 is low. Further, by maintaining the state where the inlet 11 of the container unit 10 is disposed in the vicinity of the liquid surface of the liquid 60, adhesion of the biological substance 61 to the outer wall of the container unit 10 can be suppressed.

  In addition, in Embodiment 1, although the container part 10 demonstrated the structure arrange | positioned so that the axial direction of the container part 10 may become the same as a perpendicular direction, it is not limited to this. For example, the container unit 10 may be arranged such that the axial direction of the container unit 10 is oblique to the vertical direction. As described above, the filtration device 100A may be disposed obliquely to perform filtration.

  In Embodiment 1, although the container part 10 demonstrated the structure which is a cylindrical body, it is not limited to this. The container part 10 should just be the structure which introduce | transduces the liquid 60 into an inside, can be discharged | emitted through the film part 20, and can discharge the liquid 60, for example, may be a shape of polygonal columns, such as a square pole.

  In Embodiment 1, although the film part 20 uses the metal thin film, it is not limited to this. The membrane unit 20 may be a porous membrane, and may be, for example, a membrane, filter paper, non-woven fabric, or the like.

  In Embodiment 1, although the film part 20 is provided in the inlet 11 of the container part 10, it is not limited to this. The film part 20 should just pass the liquid 60 from 1st main surface VS1 to 2nd main surface VS2 located in a perpendicular direction upper side from 1st main surface VS1, and the position of the film part 20 is not limited. For example, the membrane unit 20 may be provided between the inlet 11 and the outlet 12 of the container 10. The film unit 20 may be provided at the discharge port 12. Moreover, in Embodiment 1, although the example in which one film part 20 was provided in the inlet 11 of the container part 10 was demonstrated, it is not limited to this. A plurality of membrane units 20 may be provided between the inlet 11 and the outlet 12 of the container unit 10. For example, the first film unit 20 may be provided at the inlet 11 of the container unit 10, and the second film unit 20 may be provided above the first film unit 20 in the vertical direction. Thus, the recovery rate can be improved by configuring the multistage filtration device 100A using the plurality of membrane units 20. In addition, by designing the size of the through hole of the first membrane unit 20 and the size of the through hole of the second membrane unit 20 to be different, cells of two different sizes can be filtered. This allows the cells to be classified.

  In Embodiment 1, although the film part 20 is arrange | positioned so that 1st main surface VS1 and 2nd main surface VS2 may become perpendicular | vertical with respect to the perpendicular direction, it is not limited to this. FIG. 11 shows a filtration device 100B according to a modification of the first embodiment. As in the filtration device 100B shown in FIG. 11, the membrane unit 20 may be disposed obliquely with a predetermined angle θ with respect to the vertical direction. In other words, the film unit 20 may be arranged to be inclined with respect to the horizontal direction. In addition, a part of the film unit 20 may be disposed obliquely with respect to the vertical direction. With such a configuration, the allowable amount of the cake layer formed in the film unit 20 is increased, and therefore clogging of the film unit 20 by the biological material 61 can be further suppressed. That is, by disposing the membrane unit 20 obliquely, it is possible to further increase the area in which the membrane unit 20 contacts the liquid 60, and the filtration efficiency is improved.

  Although the vent 31 is formed by providing a gap between the outer wall of the container portion 10 and the inner wall of the liquid container 30 in the first embodiment, the present invention is not limited to this. For example, the vent 31 may be formed by providing a hole in the side wall of the liquid container 30. Further, a ventilation filter may be provided instead of the ventilation port 31.

  In Embodiment 1, although the liquid container 30 demonstrated the structure provided with the vent 31 which relieves | hangs the pressure concerning a liquid, it is not limited to this. For example, the liquid container 30 may not have the vent 31 and may not open the liquid 60 to the outside.

  In Embodiment 1, although the structure which makes the film | membrane part 20 pass the liquid 60 was demonstrated, when the attraction | suction part 40 aspirates the liquid 60 in the liquid container 30, it is not limited to this. For example, the liquid 60 may be passed from the first main surface VS1 to the second main surface VS2 of the film unit 20 by pressurizing the liquid 60 in the liquid container 30 from a position corresponding to the vent 31. Alternatively, the liquid 60 may be passed from the first main surface VS1 to the second main surface VS2 of the membrane unit 20 by applying a centrifugal force to the liquid 60 in the liquid container 30 using a centrifuge.

  In Embodiment 1, although suction part 40 explained composition which is a syringe, it is not limited to this. For example, the suction unit 40 may be a pump or the like, as long as it can suction the liquid 60.

  In the first embodiment, the liquid container 30, the suction unit 40, and the moving unit 50 are not essential components, and these elements may be omitted or replaced with other elements.

  In Embodiment 1, although the example used for filtration of living thing derived substance 61 was explained about filtration device 100A, it is not limited to this. For example, filtration device 100A may be used to replace the medium in the culture vessel in which the cells are cultured. Even when the medium is exchanged during cell culture using the filtration device 100A, clogging of the membrane unit 20 by cells can be suppressed. For example, when the culture medium of the adherent cells is exchanged, the adherent cells themselves are attracted to the inner wall of the culture vessel, which makes it difficult to contact the membrane unit 20. For this reason, according to the filtration device 100A, it is possible to achieve more efficient medium exchange of the adhesive cells as compared with the filtration device of the reference example.

Second Embodiment
[overall structure]
A filtration device 100C according to a second embodiment of the present invention will be described with reference to FIG.
FIG. 12 shows a schematic configuration of a filtration device 100C of the second embodiment. In the second embodiment, points different from the first embodiment will be mainly described. In the second embodiment, the same or similar configuration as that of the first embodiment is described with the same reference numerals or similar reference numerals. Further, in the second embodiment, the description overlapping with the first embodiment is omitted.

  As shown in FIG. 12, in comparison with the filtration device 100A of the first embodiment, the filtration device 100C of the second embodiment has the first inlet 11a for introducing the liquid 60 more than the outlet 12a for discharging the liquid 60. The difference is that the container portion 10a has a curved shape so as to be located vertically upward. In addition, the filtration device 100C does not include the liquid container 30, the suction unit 40, and the moving unit 50.

  The filtration apparatus 100C includes a container unit 10a for introducing the liquid 60 containing the biological material 61, and a membrane unit 20a for separating the biological material 61 from the liquid 60.

  The container portion 10a has a first inlet 11a for introducing the liquid 60 and an outlet 12a for discharging the liquid 60, and the first inlet 11a is curved so as to be positioned above the outlet 12a in the vertical direction. It has a shape. In the second embodiment, the container portion 10a has a U-shaped tubular shape. Specifically, the bottoms of the two tubes are connected by a U-shaped curved tube and have an integrally formed shape.

  In the second embodiment, the container portion 10a has a first inlet 11a for introducing the liquid 60 at one end and a second inlet 13 for introducing washing water, and an outlet for discharging the liquid 60 at the other end. 12a and an insertion port 14 for inserting a pipette 70. Specifically, the first inlet 11a is provided at one end of the container 10a. The second inlet 13 is provided in a pipe extending upward and provided on the side wall at one end side of the container portion 10a. The discharge port 12a is provided in a horizontally extending pipe provided on the side wall on the other end side of the container portion 10a. The insertion port 14 is provided at the other end of the container portion 10a. The first inlet 11a and the second inlet 13 at one end of the container portion 10a are provided vertically above the outlet 12a at the other end.

  In the second embodiment, the film portion 20a is a metallic thin film having a plurality of through holes 21 and is disposed between the first inlet 11a and the outlet 12a. Specifically, the film portion 20a is disposed at the bottom of the tube on the other end side of the container portion 10a and at the connection portion between the tube on the other end side and the U-curved tube. The first main surface VS1 of the film unit 20a is located below the second main surface VS2 in the vertical direction.

[Operation of filtration device]
The operation of the filtration device 100C will be described using FIGS. 13A to 13D.
13A to 13C show the operation of filtration in the filtration device 100C. FIG. 13D shows the operation of washing the biological substance 61 in the filtration device 100C.

  As shown to FIG. 13A, the liquid 60 containing the biological origin substance 61 is introduce | transduced into the container part 10a from the 1st inlet 11a (arrow 83 shown to FIG. 13A). The liquid 60 introduced into the container portion 10a flows toward the discharge port 12a by the weight of the liquid 60 (arrow 85 shown in FIG. 13A). Specifically, the liquid level of the liquid 60 located at one end of the container portion 10a is higher than the liquid level of the liquid 60 located at the other end by a predetermined distance h. The liquid 60 introduced from the port 11a flows in the direction of gravity by its own weight. The liquid 60 having flowed in the direction of gravity flows through the curved portion in the container portion 10a and is discharged from the discharge port 12a while maintaining the force. At this time, the liquid 60 flows from the first main surface VS1 of the film portion 20a to the second main surface VS2 located vertically above the first main surface VS1 (arrow 84 shown in FIG. 13A). Therefore, the biological substance 61 can be captured on the first main surface VS1 of the film portion 20a.

  As shown in FIG. 13B, when the capture amount of the biological material 61 is increased on the first main surface VS1 of the film part 20a, a cake layer is formed by the captured biological material 61.

  As shown in FIG. 13C, when a cake layer is formed in the membrane part 20a, the pipette 70 is inserted from the insertion port 14. The liquid 60 is injected from the pipette 70 inserted into the insertion port 14 toward the membrane section 20a (arrow 86 shown in FIG. 13C). The liquid 60 from the pipette 70 flows from the second major surface VS2 of the membrane unit 20 toward the first major surface VS1. Therefore, the liquid 60 that has passed through the film unit 20a is pushed down from the second main surface VS2 of the film unit 20a toward the first main surface VS1 (arrow 87 shown in FIG. 13C). The cake layer can be removed by separating the biological material 61 trapped on the first major surface VS1 of the film portion 20a from the first major surface VS1. After the cake layer is removed, the filtration device 100C can continue the filtration by removing the pipette 70 from the insertion port 14 and introducing the liquid 60 containing the biological substance 61 from the first inlet 11a.

  Thus, the filtration device 100C filters the liquid 60 containing the biological material 61 by performing the operations shown in FIGS. 13A to 13C. Moreover, 100 C of filtration apparatuses can also concentrate the liquid 60 by repeatedly performing the operation | movement shown to FIG. 13A-FIG. 13C.

  After filtration or concentration of the liquid 60, as shown in FIG. 13D, the filtration device 100C can wash the filtered biological material 61 by introducing wash water from the second inlet 13. When washing the filtered biological material 61, the introduction of the liquid 60 from the first inlet 11a is stopped, and the washing water is introduced from the second inlet 13 (arrow 88 shown in FIG. 13D). The washing water introduced from the second inlet 13 passes through the inside of the container 10 and is discharged from the outlet 12a (arrow 85 shown in FIG. 13D). In addition, when clogging with the biological material 61 occurs in the membrane unit 20a, the pipette 70 is inserted into the insertion port 14, and washing water is injected from the pipette 70 toward the membrane unit 20a (arrow 89 shown in FIG. 13D). . After the clogging is eliminated, the pipette 70 is taken out from the insertion port 14 and washing water is introduced from the second inlet 13.

  Thus, the filtration device 100C can wash the filtered biological material 61 by performing the operation shown in FIG. 13D.

[effect]
According to the filtration device 100C according to the second embodiment, the following effects can be achieved.

  In the filtration device 100C, the container portion 10a has a U-shaped tubular shape in which bottom portions of two tubes are connected by a U-shaped curved tube. In the container portion 10a, the first inlet 11a for introducing the liquid 60 provided on one end side is positioned vertically above the discharge port 12a for discharging the liquid 60 provided on the other end side. In addition, the film unit 20a is disposed between the first inlet 11a of the container unit 10a and the discharge opening 12, and the second main located vertically above the first main surface VS1 from the first main surface VS1. The liquid 60 is allowed to pass through the surface VS2. With such a configuration, the liquid 60 is allowed to pass from the first main surface VS1 to the second main surface of the film portion 20a to the second main surface VS2 by the weight of the liquid 60 introduced into the first inlet 11a. 61 can be separated.

  In the filtration device 100C, an insertion port 14 for inserting the pipette 70 is provided on the other end side of the container portion 10a. With such a configuration, when clogging occurs in the membrane portion 20a, the pipette 70 is inserted into the insertion port 14, and liquid from the pipette 70 toward the first main surface VS1 from the second main surface VS2 of the membrane portion 20a 60 can be injected. The liquid 60 injected from the pipette 70 is formed on the first main surface VS1 of the membrane portion 20a by pushing back the biological material 61 blocking the through hole 21 of the membrane portion 20a to the first inlet 11a side. The cake layer can be removed.

  In addition, the pressure due to the injection of the liquid 60 from the pipette 70 is not so large as to cause the deformation of the biological substance 61. Therefore, in the filtration device 100C, clogging of the membrane portion 20a can be resolved without damaging the biological material 61 by using the pipette 70. Further, by repeatedly injecting (pipetting) the liquid 60 with the pipette 70, it is possible to maintain a state in which clogging does not occur in the membrane unit 20a.

  In the filtration device 100C, a second inlet 13 for introducing washing water is provided on one end side of the container portion 10a. With such a configuration, by introducing washing water from the second inlet 13 after filtration or concentration of the liquid 60, the biological substance 61 captured by the membrane unit 20a can be easily washed.

  In the second embodiment, the configuration in which the second inlet 13 for introducing washing water to one end and the insertion port 14 for inserting the pipette 70 to the other end has been described, but the present invention is not limited thereto. For example, the configuration may be such that washing water is introduced from the first inlet 11 a without providing the second inlet 13. Alternatively, the pipette 70 may be inserted into the discharge port 12 a without providing the insertion port 14. Such a configuration makes it possible to filter the liquid 60 with a simple configuration.

  In Embodiment 2, although the structure which introduce | transduces wash water from the 2nd inlet 13 was demonstrated, it is not limited to this. For example, a liquid (substituting liquid) different from the liquid 60 may be introduced from the second inlet 13 and the liquid 60 in the container unit 10 may be substituted with another liquid.

  In the second embodiment, the pipette 70 inserted into the insertion port 14 injects the liquid 60 toward the membrane portion 20a in order to remove the cake layer formed on the first main surface VS1 of the membrane portion 20a. Although explained, it is not limited to this. For example, the pipette 70 may be used to agitate the liquid 60 in the container portion 10a. In addition, the pipette 70 may introduce a substitution solution toward the membrane unit 20a and replace the liquid 60 in the container unit 10 with another liquid. The pipette 70 may perform pipetting in a closed system in which the injection pressure of the liquid 60 from the pipette 70 does not escape out of the container portion 10 a by inserting the pipette 70 so as to close the insertion port 14.

  In the second embodiment, the container portion 10a has been described as a U-shaped tube, but is not limited thereto. For example, a curved U-shaped portion of the container portion 10a may use a rubber pipe to provide flexibility and a height difference between the left and right pipes, and may be provided with a three-way branch for recovery. With such a configuration, the biological substance 61 can be easily recovered.

  In Embodiment 2, although the structure which filters with the dead weight of the liquid 60 introduce | transduced from the 1st inlet 11a was demonstrated, it is not limited to this. For example, filtration may be performed by pressurizing the liquid 60 from the first inlet 11a. With such a configuration, filtration can be performed in a short time.

  Although the example using the pipette 70 has been described in the second embodiment, the pipette 70 is not an essential component. For example, the pipette 70 may be omitted or replaced with another element such as gas pressurization.

Third Embodiment
[overall structure]
A filtration device 100D according to a third embodiment of the present invention will be described with reference to FIG.
FIG. 14 shows a schematic configuration of a filtration device 100D of the third embodiment. In the third embodiment, points different from the first embodiment will be mainly described. In the third embodiment, the same or similar configuration as that of the first embodiment is described with the same reference numerals or similar reference numerals. Further, in the third embodiment, the description overlapping with the first embodiment is omitted.

  As shown in FIG. 14, the filtration device 100D of the third embodiment has a configuration in which the container portion 10b has a configuration capable of moving the inner wall of the liquid container 30b, compared to the filtration device 100A of the first embodiment. It is different. Further, filtration device 100D is different from filtration device 100A in that membrane portion 20b is provided obliquely to first side wall 10ba of container portion 10b. The filtration device 100D does not include the suction unit 40.

  The filtering device 100D is disposed between the inlet 11b and the outlet 12b, the container portion 10b having the inlet 11b for introducing the liquid 60 containing the biological substance 61 and the outlet 12b for discharging the liquid 60, A film unit 20 b having a plurality of through holes 21 and a liquid container 30 b for holding a liquid 60 are provided. The container portion 10b is attached to the inside of the liquid container 30b, and can move inside the liquid container 30b. The filtration device 100D moves the container portion 10b downward to move the liquid 60 in the liquid container 30b from the first main surface VS1 of the film portion 20b to the second main surface vertically above the first main surface VS1. I am passing it to VS2.

  In the third embodiment, the container portion 10b includes a first side wall 10ba disposed in the liquid container 30, a second side wall 10bb disposed outside the liquid container 30, and a first side wall 10ba and a second side wall 10bb. A third side wall 10 bc to be connected is provided.

  The first side wall 10ba is an inner cylinder disposed in the liquid container 30 and movable toward the liquid 60 held in the liquid container 30b. At one end of the first side wall 10ba, an introduction port 11b into which the liquid 60 in the liquid container 30b is introduced is provided, and at the other end, a discharge port 12b from which the liquid 60 is discharged is provided. The inlet 11b is positioned below the outlet 12b in the vertical direction. The film part 20b which has several through-holes 21 is arrange | positioned at the inlet 11b.

  In the first side wall 10ba, the outer peripheral portion of the film portion 20b is designed with a material and dimensions capable of sliding the inner wall of the liquid container 30b. That is, the outer peripheral portion of the membrane portion 20b is designed such that the biological substance 61 does not pass between the outer peripheral portion of the membrane portion 20b and the liquid container 30b, and the membrane portion 20b faces the liquid 60 in the liquid container 30b. It is designed to be movable.

  The second side wall 10bb is an outer cylinder which is disposed outside the liquid container 30 and can slide on the outer wall of the liquid container 30. The second side wall 10bb includes a first moving unit 50a on the inner wall. The first moving unit 50 a engages with the second moving unit 50 b provided on the outer wall of the liquid container 30 and moves the container unit 10 b toward the liquid 60. For example, the first moving portion 50a is formed by an internal thread and is screwed with the second moving portion 50b formed by an external thread.

  The third side wall 10bc is a ring-shaped member connecting the first side wall 10ba and the second side wall 10bb so as to form a space in which the side wall of the liquid container 30 enters between the first side wall 10ba and the second side wall 10bb. is there. Specifically, the third side wall 10 bc connects the end of the first side wall 10 ba on the outlet 12 b side and the upper end of the second side wall 10 bb. The first side wall 10ba, the second side wall 10bb, and the third side wall 10bc may be integrally formed.

  The film part 20b is arrange | positioned at the inlet 11b of 1st side wall 10ba, has predetermined angle (theta) with respect to the inner wall of 1st side wall 10ba, and is arrange | positioned diagonally. That is, the film portion 20b is disposed to be inclined with respect to the horizontal direction. The hole diameter of the through hole 21 of the membrane portion 20b is designed to be a size that allows the liquid 60 to pass through without passing through the biological substance 61 to be filtered.

  The liquid container 30 b is a cylindrical container that holds the liquid 60 containing the biological substance 61. The first side wall 10ba of the container portion 10b is slidably disposed on the inner wall of the liquid container 30b. The first side wall 10ba is movable downward toward the liquid 60 held in the liquid container 30b. The outer wall of the liquid container 30b includes a second moving unit 50b. The second moving unit 50b engages with the first moving unit 50a provided on the second side wall 10bb of the container unit 10b. For example, the second moving portion 50b is formed by an external thread as described above, and engages with the first moving portion 50a formed by an internal thread provided on the second side wall 10bb of the container portion 10b.

[Operation of filtration device]
The operation of the filtration device 100D will be described with reference to FIGS. 15A and 15B.
15A and 15B show the operation of the filtration in the filtration device 100D.

  As shown in FIG. 15A, the container portion 10b is removed from the liquid container 30b, and the liquid 60 containing the biological material 61 is introduced into the liquid container 30.

  As shown in FIG. 15B, the container portion 10b is attached to the liquid container 30b. Specifically, the first side wall 10ba of the container portion 10b is disposed inside the liquid container 30b, and the first moving portion 50a of the second side wall 10bb of the container portion 10b and the second moving portion 50b of the liquid container 30b are engaged. Match. Then, by rotating the container portion 10b in the circumferential direction 90, the container portion 10b moves vertically downward toward the liquid 60 held in the liquid container 30b (arrow 91 shown in FIG. 15B). That is, the film unit 20 b disposed at the inlet 11 b of the container unit 10 b moves vertically downward toward the liquid 60. For this reason, in the film part 20b, the liquid 60 in the liquid container 30b passes from the first main surface VS1 to the second main surface VS2 of the film part 20b (arrow 92 shown in FIG. 15B).

  In the third embodiment, the film portion 20b provided obliquely to the inner wall of the first side wall 10ba of the container portion 10b moves toward the liquid 60 while rotating about the axis of the container portion 10b. Therefore, the biological material 61 attached to the first main surface VS1 of the film portion 20b can be peeled off by vibration due to rotation.

  In the filtration device 100D, after the filtration is completed, for example, the liquid 60 is discharged from the outlet 12b of the container portion 10b by turning the filtration device 100D upside down. Thereafter, a new liquid or replacement liquid may be introduced. Alternatively, the liquid 60 may be discharged from the discharge port 12 b by a method such as suction.

[effect]
According to filtration device 100D according to the third embodiment, the following effects can be achieved.

  In the filtration device 100D, the film unit 20b is moved toward the liquid 60 held in the liquid container 30b by moving the container unit 10b downward. Therefore, the liquid 60 in the liquid container 30b passes through the second main surface VS2 located vertically above the first main surface VS1 from the first main surface VS1 of the film unit 20b. With such a configuration, the biological substance 61 can be easily separated from the liquid 60 by gradually moving the membrane portion 20 b toward the liquid 60.

  In the filtration device 100D, the liquid 60 extracted inside the container portion 10b can be discharged from the outlet 12b by turning the filtration device 100D upside down after the filtration is completed. In addition, after discharging the liquid 60, by introducing a new liquid or a replacement liquid, cleaning, liquid exchange, etc. can be easily performed. As described above, in the filtration device 100D, pipetting by a pipette or the like is unnecessary, and the usability for the user is improved.

  In the filtration device 100D, the membrane unit 20b is disposed obliquely to the inner wall of the first side wall 10ba of the container unit 10b. Therefore, when the film unit 20b is moved toward the liquid 60 in the liquid container 30b, adhesion of the biological substance 61 to the first main surface VS1 of the film unit 20b can be suppressed. Thereby, clogging due to the adhesion of the biological material 61 to the film portion 20b can be suppressed.

  In the filtration device 100D, the membrane unit 20b moves downward while rotating. For this reason, the biological origin substance 61 adhering to 1st main surface VS1 of the film part 20b can be peeled off by the vibration which arose by rotation of the film part 20b. Thereby, clogging due to the adhesion of the biological material 61 to the film portion 20b can be suppressed.

  In Embodiment 3, although the film part 20b demonstrated the structure provided diagonally with respect to the inner wall of 1st side wall 10ba of the container part 10b, it is not limited to this. For example, a part of the film portion 20b may be provided obliquely to the inner wall of the first side wall 10ba. The film portion 20b may be provided perpendicularly to the inner wall of the first side wall 10ba.

  In the third embodiment, the configuration including the first moving unit 50a provided on the inner wall of the second side wall 10bb of the container unit 10b and the second moving unit 50b provided on the outer wall of the liquid container 30 has been described. It is not limited to this. For example, the filtration device 100D directs the membrane unit 20b to the liquid 60 held in the liquid container 30 by the weight of the membrane unit 20b and the container unit 10b without providing the first moving unit 50a and the second moving unit 50b. You may move it. Alternatively, the filtration device 100D may move the membrane unit 20b toward the liquid 60 by pushing the third side wall 10bc downward.

  In the third embodiment, the second liquid is introduced to the inside of the container portion 10b so that the liquid 60 can easily pass through the through hole 21 of the film portion 20b in a state where the container portion 10b is attached in the liquid container 30b. In the condition, filtration may be started. That is, filtration device 100D may start filtration in the state where the 2nd fluid is held on the 2nd principal surface VS2 of membrane part 20b.

  In the third embodiment, a stirring blade for stirring the liquid 60 may be provided inside the liquid container 30b. The stirring blade can suppress adhesion of the biological substance 61 in the liquid 60 to the film portion 20b.

Embodiment 4
[overall structure]
A filtration device 100E according to a fourth embodiment of the present invention will be described with reference to FIG.
FIG. 16 shows a schematic configuration of a filtration device 100E of the fourth embodiment. In the fourth embodiment, points different from the first embodiment will be mainly described. In the fourth embodiment, the same or similar configuration as that of the first embodiment is described with the same reference numeral or similar reference numeral. Further, in the fourth embodiment, the description overlapping with the first embodiment is omitted.

  As shown in FIG. 16, compared with the filtration device 100A of the first embodiment, the filtration device 100E of the fourth embodiment applies a centrifugal force to the inlet 11c from the discharge port 12c of the container portion 10c, and thereby the liquid 60 is obtained. The point which has the structure which can isolate | separated the biological origin substance 61 from this. Further, the filtration device 100E does not include the suction unit 40 and the moving unit 50.

  The filtering device 100E is disposed between the inlet 11c and the outlet 12c, and the container portion 10c having the inlet 11c for introducing the liquid 60 containing the biological substance 61 and the outlet 12c for discharging the liquid 60, A film unit 20 c having a plurality of through holes 21 and a liquid container 30 c for holding a liquid 60 are provided. The container portion 10c is attached to the inside of the liquid container 30c. The filtration apparatus 100E applies the centrifugal force in the direction from the outlet 12c of the container portion 10c to the inlet 11c, thereby making the liquid 60 in the liquid container 30c from the first main surface VS1 of the film portion 20c to the first main surface VS1. It is made to pass to the 2nd principal surface VS2 located in the perpendicular direction upper side. That is, when the centrifugal force is applied, the liquid 60 flows in the direction opposite to the direction in which the centrifugal force is applied in the membrane portion 20c. Further, the container portion 10c is provided with a hole for degassing.

  In the fourth embodiment, the container unit 10c is disposed in the liquid container 30c. With the container portion 10c disposed in the liquid container 30c, the lid 32 covering the opening of the liquid container 30c is attached.

[Operation of filtration device]
The operation of the filtration device 100E will be described using FIGS. 17A to 17F.
17A-17F show the operation of filtration in the filtration device 100E.

  As shown to FIG. 17A, the container part 10c is removed from the liquid container 30c, and the liquid 60 containing the biological origin substance 61 is introduced in the liquid container 30c.

  As shown in FIG. 17B, the container portion 10c is mounted in the liquid container 30c holding the liquid 60. In the fourth embodiment, the lid 32 is attached while holding the container portion 10c inside the liquid container 30c. That is, the inlet 11c of the container portion 10c is disposed in the liquid 60 in the liquid container 30c. The attachment of the lid 32 is performed, for example, by screwing an internal thread provided inside the lid 32 and an external thread provided on the outer wall of the liquid container 30c.

  By attaching the container portion 10c to the inside of the liquid container 30c, the liquid 60 held in the liquid container 30c is pushed up between the outer wall of the container portion 10c and the inner wall of the liquid container 30c. In this state, the filtration device 100E is set in the centrifuge.

  As shown in FIG. 17C, when a centrifugal force (arrow 93 shown in FIG. 17C) is applied from the outlet 12c of the container portion 10c to the inlet 11c by a centrifuge (not shown), the outer wall of the container portion 10c and The liquid 60 pushed up with the inner wall of the liquid container 30c flows toward the bottom of the liquid container 30c (arrow 94 shown in FIG. 17C). At this time, the biological substance 61 collects at the bottom of the liquid container 30c. On the other hand, the liquid 60 passes from the first main surface VS1 of the film portion 20c to the second main surface VS2 located vertically above the first main surface VS1 to the discharge port 12c from the inlet 11c of the container portion 10c. Flow toward (arrow 95 shown in FIG. 17C). The height of the liquid surface of the liquid 60 between the outer wall of the container portion 10c and the inner wall of the liquid container 30c is the same as the height of the liquid surface of the liquid 60 which has passed through the membrane portion 20c. The height of the liquid surface of the liquid 60 between the outer wall of the container portion 10c and the inner wall of the liquid container 30c and the height of the liquid surface of the liquid 60 which has passed through the membrane portion 20c are the liquid containers 30c at the respective places. Means the height of the liquid 60 from the bottom of the.

  As shown in FIG. 17D, the centrifuge is stopped and the lid 32 is removed from the fluid container 30c. After removing the lid 32, the liquid 60 which has passed through the membrane unit 20c is discharged. Discharge of the liquid 60 is performed by turning the liquid container 30c upside down with the container portion 10c attached to the liquid container 30c. At this time, the biological material 61 accumulated at the bottom of the liquid container 30c can not pass through the through hole 21 of the film portion 20c. Therefore, the liquid 60 which does not contain the biological substance 61 is discharged from the discharge port 12 c.

  As shown in FIG. 17E, the lid 32 is attached to the liquid container 30c, and a centrifugal force from the outlet 12c to the inlet 11c is applied to the filtration device 100E by a centrifugal separator (arrow 93 shown in FIG. 17E). When centrifugal force is applied to the filtering device 100E, the liquid 60 between the outer wall of the container portion 10c and the inner wall of the liquid container 30c flows toward the bottom of the liquid container 30c as in the operation shown in FIG. 17C (FIG. 17E Arrows 94). Then, the liquid 60 containing no biological substance 61 flows from the bottom of the liquid container 30c to the inlet 11c of the container 10c and passes through the membrane 20c (arrow 95 shown in FIG. 17E). The height of the liquid surface of the liquid 60 between the outer wall of the container portion 10c and the inner wall of the liquid container 30c is the same as the height of the liquid surface of the liquid 60 which has passed through the membrane portion 20c.

  As shown in FIG. 17F, the centrifuge is stopped and the lid 32 is removed from the fluid container 30c. Thereafter, in the same manner as the operation shown in FIG. 17D, the liquid 60 which has passed through the film portion 20c and accumulated inside the container portion 10c is discharged from the discharge port 12c.

  Thus, the filtration apparatus 100E separates the biological substance 61 from the liquid 60 using centrifugal force by performing the operations shown in FIGS. 17A to 17F. Furthermore, the filtration apparatus 100E can concentrate the biological substance 61 by repeating the operations shown in FIGS. 17A to 17F. In addition, after concentration, the filtration device 100E can introduce washing water to wash the biological substance 61. Further, in the filtration device 100E, the liquid 60 can be replaced with another fluid by separating the biological material 61 and introducing a replacement fluid.

[effect]
According to the filtration device 100E in the fourth embodiment, the following effects can be obtained.

  In the filtration device 100E, when the centrifugal force is applied, the liquid 60 is allowed to pass in the opposite direction to the direction in which the centrifugal force is applied in the membrane unit 20c. Specifically, when the filtration device 100C applies a centrifugal force, the height of the liquid surface of the liquid 60 between the outer wall of the container portion 10c and the inner wall of the liquid container 30c and the amount of the liquid 60 passing through the membrane portion 20c. The liquid 60 is vertically vertically passed through the film portion 20c by utilizing the height difference with the height of the liquid surface. With such a configuration, the biological substance 61 can be separated from the liquid 60 without clogging of the membrane part 20 c with the biological substance 61. Moreover, in the filtration apparatus 100E, concentration of the biological substance 61 can be easily performed by repeatedly performing filtration by centrifugation.

  In the filtering device 100E, when taking out the liquid 60 that has passed through the membrane section 20c, for example, the liquid 60 can be easily discharged by turning the filtering device 100E upside down. As described above, in the filtering device 100E, pipetting by the operator is unnecessary, and the liquid 60 can be discharged without requiring the operator to have a special skill. As a result, in the filtration device 100E, a decrease in the recovery rate of the biological substance 61 due to pipetting does not occur.

  In the filtration device 100E, after the biological substance 61 is separated, washing water is introduced and centrifugation is performed, whereby the biological substance 61 can be washed. In addition, in the filtration device 100E, replacement of the liquid can be easily performed by separating the biological material 61 and introducing a replacement solution.

  In addition, in Embodiment 4, although the structure provided with the lid | cover 32 was demonstrated, it is not limited to this. The lid 32 is not an essential component, and may be omitted or replaced with another element.

  While the present invention has been fully described in connection with the preferred embodiments with reference to the accompanying drawings, various changes and modifications will be apparent to those skilled in the art. Such variations and modifications are to be understood as being included therein without departing from the scope of the present invention as set forth in the appended claims.

  The present invention is an invention relating to a filtration apparatus and a filtration method, and is excellent in the point of filtering a high concentration liquid and the point of filtering in a short time. For example, it is useful in the fields of chemical analysis, drug loading / pharmaceutics, clinical examination, public health management, environmental measurement and the like.

DESCRIPTION OF SYMBOLS 10 container part 11 inlet 12 discharge port 20 membrane part 21 through-hole 30 liquid container 31 vent 32 lid 40 suction part 41 tube 50 moving part 60 liquid 61 biological material 70 pipette 100 filtration device

Claims (11)

A container portion having an inlet for introducing a liquid containing a biological substance and an outlet for discharging the liquid;
A membrane portion provided between the inlet and the outlet of the container portion and having a plurality of through holes;
A liquid container for holding the liquid;
Equipped with
The inlet of the container portion is disposed in the liquid held in the liquid container,
The film portion has a first main surface and a second main surface located above the first main surface, and moves toward the liquid while rotating inside the liquid container.
By passing the liquid from the first major surface of the front Kimaku portion to said second main surface, for separating the biological material from the liquid filtering device. The container portion has a first side wall disposed inside the liquid container,
The film portion is provided inside the first side wall,
The first side wall moves toward the liquid while rotating.
The filtration device according to claim 1. The container portion has a second side wall disposed outside the liquid container and connected to the first side wall,
The second side wall has a first moving portion on an inner wall,
The liquid container has a second moving part on an outer wall,
The first moving unit engages with the second moving unit and moves the container portion toward the liquid while rotating the container portion with respect to the liquid container.
The filtration device according to claim 2. The first moving part is an internal thread,
The second moving unit is a male screw,
The filtration device according to claim 3, wherein the first moving unit is screwed with the second moving unit. The filtration device according to any one of claims 1 to 4, wherein the liquid container comprises a stirring blade for stirring the liquid. The filtration device according to any one of claims 1 to 5 , wherein the membrane part is a metal thin film. The filtration device according to any one of claims 1 to 6, wherein at least a part of the membrane part is provided obliquely with respect to the vertical direction. A method of filtering a biological substance from a liquid, comprising
A container unit including an inlet for introducing a liquid containing a biological material and an outlet for discharging the liquid, and a plurality of through holes provided between the inlet and the outlet of the container. Preparing a filtration device comprising a membrane part and a liquid container for holding the liquid,
Separating the bio-derived material by passing the liquid from a first major surface of the membrane portion to a second major surface located above the first major surface;
Including
The step of separating includes moving the membrane toward the liquid while rotating the membrane inside the liquid container . Wherein the step of separating at least part of Ru passed through the liquid to the membrane portion provided obliquely with respect to the vertical direction, the method of filtration according to claim 8. The filtration method according to claim 8, further comprising the step of discharging the liquid from the outlet of the container portion by inverting the filtration device. The filtration method according to any one of claims 8 to 10, further comprising the step of introducing a liquid into the inside of the container portion.

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