WO2019059241A1 - 濾過装置、濾過システム及び濾過方法 - Google Patents
濾過装置、濾過システム及び濾過方法 Download PDFInfo
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- WO2019059241A1 WO2019059241A1 PCT/JP2018/034688 JP2018034688W WO2019059241A1 WO 2019059241 A1 WO2019059241 A1 WO 2019059241A1 JP 2018034688 W JP2018034688 W JP 2018034688W WO 2019059241 A1 WO2019059241 A1 WO 2019059241A1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
- B01D61/18—Apparatus therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
- B01D61/147—Microfiltration
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
- B01D61/20—Accessories; Auxiliary operations
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
- B01D61/22—Controlling or regulating
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D63/00—Apparatus in general for separation processes using semi-permeable membranes
- B01D63/08—Flat membrane modules
- B01D63/087—Single membrane modules
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
- C12M3/06—Tissue, human, animal or plant cell, or virus culture apparatus with filtration, ultrafiltration, inverse osmosis or dialysis means
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2313/00—Details relating to membrane modules or apparatus
- B01D2313/08—Flow guidance means within the module or the apparatus
- B01D2313/086—Meandering flow path over the membrane
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2313/00—Details relating to membrane modules or apparatus
- B01D2313/10—Specific supply elements
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2313/00—Details relating to membrane modules or apparatus
- B01D2313/12—Specific discharge elements
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2313/00—Details relating to membrane modules or apparatus
- B01D2313/18—Specific valves
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2315/00—Details relating to the membrane module operation
- B01D2315/10—Cross-flow filtration
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2315/00—Details relating to the membrane module operation
- B01D2315/18—Time sequence of one or more process steps carried out periodically within one apparatus
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2321/00—Details relating to membrane cleaning, regeneration, sterilization or to the prevention of fouling
- B01D2321/20—By influencing the flow
- B01D2321/2033—By influencing the flow dynamically
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2321/00—Details relating to membrane cleaning, regeneration, sterilization or to the prevention of fouling
- B01D2321/20—By influencing the flow
- B01D2321/2083—By reversing the flow
Definitions
- the disclosed technology relates to a filtration device, a filtration system and a filtration method.
- the following techniques are known as techniques relating to a filtration device provided with a filtration membrane.
- JP-A-8-24590 discloses a hollow fiber membrane hollow portion of a liquid to be treated in in-out filtration using a hollow fiber membrane module in which both open ends of the hollow fiber membrane are separately focused and fixed. Feeding is alternately switched from the first fluid outlet and second fluid outlet, and immediately after the fluid flow direction is switched, filtration is performed while discharging part of the liquid to be treated from the fluid outlet on the supply stop side. It has been described that.
- JP 2009-291744 A filtration processing is carried out by a cross flow method in which the water to be treated is allowed to flow from one end side to the other end side of the membrane module along the membrane surface of the membrane module.
- a membrane filtration method with a filtration device for collecting filtered water is described.
- JP-A-2017-104832 discloses a membrane separation including a filtration membrane that divides a casing into a concentration side space to which water to be treated is supplied and a permeation side space to which permeate water separated from the water to be treated is accommodated.
- a plurality of U-shaped tubular first connecting members are provided in which one ends of a plurality of filtration membranes and the other ends are connected so that a plurality of filtration membranes are connected in series. Have been described.
- a membrane separation process is performed to remove spent medium and dead cells from the cell suspension.
- a tangential flow method also referred to as a cross flow method
- the tangential flow method is a method of sending the permeation component to the permeation side while flowing the cell suspension to be subjected to the membrane separation treatment along the membrane surface of the filtration membrane. According to the tangential flow method, it is possible to reduce the damage to cells as compared with the dead end method.
- the filtration efficiency In order to realize large-scale culture of cells, it is necessary to increase the throughput per unit time of the membrane separation process, that is, the filtration efficiency.
- As a method of enhancing the filtration efficiency it is conceivable to increase the liquid transfer rate of the cell suspension flowing on the membrane surface of the filtration membrane, but in this case, there is a concern that the cell may be damaged. Therefore, it is considered effective to expand the area of the filtration membrane as a method of enhancing the filtration efficiency. For example, by making the flow path of the cell suspension flowing on the membrane surface of the filtration membrane meandering, the area of the filtration membrane can be increased while maintaining the area efficiency.
- the membrane surface pressure difference (hereinafter referred to as filtration pressure) of the filtration membrane tends to be nonuniform.
- filtration pressure becomes uneven on the membrane surface of the filtration membrane, cell clogging occurs in a portion where the filtration pressure of the filtration membrane is relatively high, and the cell recovery rate decreases.
- a reduction in cell recovery leads to a reduction in cell culture efficiency, which in turn increases the cost of cell culture.
- One embodiment of the present invention provides a filtration device, a filtration system, and a filtration method that make it possible to increase the area of the filtration membrane while securing the uniformity of the filtration pressure on the membrane surface of the filtration membrane.
- the filtration device includes a meandering flow path, a filtration membrane that separates the supply side and the permeation side of the flow path, a first flow port provided at one end of the flow side of the flow path, and a flow path
- the second flow port provided at the other end of the supply side, the first discharge port provided at the transmission side of the flow path, and the first discharge port at the transmission side of the flow path are provided at different positions And a second outlet.
- the distance from the center of the first flow port to the farthest position of the membrane surface of the filtration membrane is W1, and the distance from the center of the first flow port to the center of the first discharge port is d1.
- W2 the distance from the center of the second flow port to the center of the second discharge port
- the flow path has a plurality of turnbacks, and the distances from one of the turnbacks to the next turn may be the same.
- One end and the other end of the flow path and each of the folds on one side are provided on a first straight line, and each of the folds on the other side is on a second straight line parallel to the first straight line May be provided.
- the flow path may have a plurality of turnbacks, and the distance from one of the turnbacks to the next turn may be short compared to the other portions.
- a filtration system performs the first liquid transfer control for controlling the liquid transfer of the liquid passing through the above-described filtration device and the first flow port and the second flow port. And a control unit that performs second liquid transfer control that controls liquid transfer of the liquid discharged from the first discharge port and the second discharge port in conjunction with the control.
- the control unit performs, as the first liquid transfer control, the first liquid transfer for discharging the liquid flowed in from the first flow port from the second flow port, and the liquid flowed in from the second flow port.
- the second liquid transfer to be discharged from the first flow port may be alternately performed, and as the second liquid transfer control, while the first liquid transfer is performed, the liquid that has permeated the filtration membrane is While discharging from the 1st outlet and carrying out the 2nd sending, the liquid which penetrated the filtration membrane may be discharged from the 2nd outlet.
- the liquid may be a cell suspension.
- a filtration method is a method of filtering a liquid using the above-described filtration device, which is a first liquid transfer in which the liquid flowed in from the first flow port is made to flow out from the second flow port; The liquid flowing in from the second flow port is alternately sent out from the first flow port and the second liquid transfer is made to flow out from the first flow port. And discharging the liquid that has permeated the filtration membrane from the second outlet while discharging the second membrane from the second outlet.
- the cell suspension may be targeted for the first and second delivery.
- a filtration device a filtration system and a filtration method which make it possible to increase the area of the filtration membrane while securing the uniformity of the filtration pressure on the membrane surface of the filtration membrane.
- FIG. 2 is a cross-sectional view taken along line 2A-2A in FIG.
- FIG. 2 is a cross-sectional view taken along line 2B-2B in FIG.
- FIG. 2 is a cross-sectional view taken along the line 2C-2C in FIG.
- FIG. 2 is a cross-sectional view taken along line 2D-2D in FIG.
- It is a top view showing an example of composition of a filtration device concerning an embodiment of art of an indication.
- FIG. 6A It is a figure which shows an example of distribution of liquid feeding speed in each position on the dashed-dotted line L2 in FIG. 6A. It is the figure which accumulated and showed the filtration pressure appropriate area
- FIG. 1 is a plan view showing an example of the configuration of a filtration device 1 according to an embodiment of the disclosed technology
- FIGS. 2A, 2B, 2C and 2D are lines 2A-2A and 2B-2B in FIG.
- FIG. 5 is a cross-sectional view taken along the lines 2C-2C and 2D-2D.
- the filtration apparatus 1 can be used as an example in the application of the membrane separation process of the cell suspension by a tangential flow system.
- the filtration device 1 includes an upper case 11 and a lower case 12, and a filtration membrane 20 provided between the upper case 11 and the lower case 12.
- the filtration device 1 also has an O-ring 13 provided along the outer edge of the filtration membrane 20 and an O-ring 14 surrounding the outer periphery of the filtration membrane 20.
- the O-ring 13 secures the adhesion between the filtration membrane 20 and the upper case 11, and the O-ring 14 secures the adhesion between the upper case 11 and the lower case 12.
- the upper case 11 and the lower case 12 are respectively provided with projections 15 and 16 which project toward the filtration membrane 20 and whose tip ends are in contact with the filtration membrane 20. , And a channel 30 through which the cell suspension passes is formed. That is, the protrusions 15 and 16 form the wall surface of the flow path 30.
- the filtration membrane 20 separates the supply side 31 and the permeation side 32 of the flow path 30.
- the flow path 30 is configured by connecting a plurality of unit flow paths 30a at a plurality of folded portions 30b. That is, the flow path 30 is meandering as shown in FIG.
- the filtration membrane 20 has a rectangular shape, and the lengths of the unit flow paths 30a are the same as one another. That is, each of one end and the other end of the flow path 30 and the folded portion 30b on one side is provided on the first straight line, and each of the folded portions 30b on the other side is parallel to the first straight line. It is provided on a second straight line. The respective distances from one of the plurality of folded portions 30 b to the next folded portion 30 b are the same as one another.
- the range of the processing accuracy error for example, about 0.05 mm
- the filtration membrane 20 has a plurality of pores of a diameter smaller than the diameter of the cells contained in the cell suspension.
- MF membrane Microfiltration Membrane
- a first flow port 41 is provided at one end of the supply side 31 of the flow path 30, and a second flow port 42 is provided at the other end of the supply side 31 of the flow path 30. That is, each of the first flow port 41 and the second flow port 42 is provided in the vicinity of the corner of the filtration membrane 20 having a rectangular shape.
- the cell suspension flows along the membrane surface of the filtration membrane 20 between the first flow port 41 and the second flow port 42 on the supply side 31 of the flow path 30.
- the cell suspension which has flowed in from the first flow port 41 flows along the membrane surface of the filtration membrane 20 along the supply side 31 of the meandering flow path 30 and flows out from the second flow port 42.
- the cell suspension flowing from the second flow port 42 flows along the membrane surface of the filtration membrane 20 along the supply side 31 of the meandering flow path 30 and flows out from the first flow port 41.
- the permeation side 32 of the flow path 30 is maintained at a lower pressure than the supply side 31, and while the cell suspension is flowing on the supply side 31 of the flow path 30, components smaller than the pore diameter of the filtration membrane 20 are Transparent to 32.
- the cells contained in the cell suspension do not permeate the filtration membrane 20, flow out from the first flow port 41 or the second flow port 42, and are collected.
- a first discharge port 51 and a second discharge port 52 for discharging the permeated liquid that has permeated through the filtration membrane 20 are provided at mutually different positions on the permeate side 32 of the flow path 30.
- the first discharge port 51 is provided at a position farthest from the first flow port 41 of the flow path 30, and the second discharge port 52 is a second flow path of the flow path 30. It is provided at a position farthest from the circulation port 42.
- the “position farthest from the first flow port 41” means a position on the flow path 30 where the linear distance from the first flow port 41 is the longest.
- a position farthest from the second flow port 42 means a position on the flow path 30 where the linear distance from the second flow port 42 is the longest.
- the first discharge port 51 may be provided in the vicinity of a position farthest from the first flow port 41, and the second discharge port 52 may be farthest from the second flow port 42. It may be provided in the vicinity of the following position. That is, as shown in FIG. 3, the first discharge port 51 is in the vicinity R1 of the corner A3 which is diagonal to the corner A1 of the filtration membrane 20 where the first flow port 41 is disposed.
- the second outlet 52 may be provided in the vicinity region R2 of the corner A4, which is diagonal to the corner A2 of the filtration membrane 20 where the second passage 42 is disposed. It may be arranged.
- the distance from the center of the first flow port 41 to the farthest position B1 of the membrane surface of the filtration membrane 20 is W1
- the distance from the center of the first flow port 41 to the first discharge port 51 is The distance to the center
- the distance from the center of the second flow port 42 to the farthest position B2 of the membrane surface of the filtration membrane 20 is W2
- the second discharge port from the center of the second flow port 42 Assuming that the distance to the center of 52 is d2, it is preferable to satisfy the following equations (1) and (2).
- step S ⁇ b> 1 the cell suspension is made to flow in from the first flow port 41, and the first liquid transfer is made to flow out from the second flow port 42.
- the cell suspension flowing from the first flow port 41 flows along the membrane surface of the filtration membrane 20 while meandering on the supply side 31 of the flow path 30. While the cell suspension is flowing on the supply side 31 of the flow path 30, components smaller than the pore size of the filtration membrane 20 contained in the cell suspension permeate to the permeation side 32. This concentrates the cell suspension.
- the concentrated cell suspension that has flowed out of the second flow port 42 is temporarily collected in a container (not shown).
- step S1 the permeated liquid that has permeated through the filtration membrane 20 is discharged from the first discharge port 51 while the first liquid transfer is performed.
- the second outlet 52 is closed so that the permeated liquid is not discharged from the second outlet 52.
- the pump is connected on the flow path communicating with the first discharge port 51, and the pump is operated to perform the first discharge. Suction may be generated at the outlet 51.
- step S2 it is determined whether or not there is an abnormality during the first liquid transfer. For example, when it is detected that the liquid transfer load in the first liquid transfer is excessive, it may be determined that there is an abnormality as clogging of cells in the filtration membrane 20 is occurring. If it is determined that there is an abnormality, the process proceeds to step S7, where it is notified that cell clogging has occurred in the filtration membrane 20. On the other hand, when it is determined that there is no abnormality, the process proceeds to step S3. Although the rough flow is described in the flowchart of FIG. 4, step S2 is repeatedly performed during the first liquid delivery in step S1. When there is no abnormality at the time of the first liquid feeding and the condition for ending the first liquid feeding is satisfied, the process proceeds to step S3. The condition under which the first liquid transfer ends is, for example, the case where all the predetermined throughputs have been processed. Alternatively, the determination in step S2 may be performed after the condition for ending the first liquid transfer is satisfied.
- step S 3 the collected cell suspension is introduced into the container through the second flow port 42, and the second liquid transfer is performed so as to flow out of the first flow port 41. That is, the flow direction of the cell suspension flowing on the supply side 31 of the flow path 30 is reversed.
- the cell suspension flowing from the second flow port 42 flows along the membrane surface of the filtration membrane 20 while meandering on the supply side of the flow path 30. While the cell suspension is flowing on the supply side 31 of the flow path 30, components smaller than the pore size of the filtration membrane 20 contained in the cell suspension permeate to the permeation side 32. This concentrates the cell suspension.
- the concentrated cell suspension that has flowed out of the first flow port 41 is temporarily collected in a container (not shown).
- step S3 the permeated liquid that has permeated through the filtration membrane 20 is discharged from the second discharge port 52 while performing the second liquid transfer.
- the first discharge port 51 is closed so that the permeated liquid is not discharged from the first discharge port 51.
- the pump is connected on the flow path communicating with the second discharge port 52, and the second discharge is operated by operating the pump. Suction may be generated at the outlet 52.
- step S4 it is determined whether or not there is an abnormality during the second liquid delivery. For example, when it is detected that the liquid transfer load in the second liquid transfer is excessive, it may be determined that there is an abnormality in the filtration membrane 20 as clogging of cells. If it is determined that there is an abnormality, the process proceeds to step S7, and it is informed that cell clogging has occurred in the filtration membrane. On the other hand, when it is determined that there is no abnormality, the process proceeds to step S5. Although the rough flow is described in the flowchart of FIG. 4, step S4 is repeatedly performed during the second liquid delivery in step S3. When there is no abnormality at the time of the second liquid feeding and the condition for ending the second liquid feeding is satisfied, the process proceeds to step S5. The condition under which the second liquid transfer ends is, for example, the case where all the predetermined throughputs have been processed. Alternatively, the determination in step S4 may be performed after the condition for ending the second liquid transfer is satisfied.
- step S5 it is determined whether or not the number of cycles of processing in which a series of processes including steps S1 and S2 described above is one unit has reached a predetermined number. If it is determined that the number of cycles in the series of processes has not reached the predetermined number, the process returns to step S1. If it is determined that the number of cycles of the series of processes has reached a predetermined number, the process proceeds to step S6.
- step S6 it is determined whether the concentration of cells in the concentrated cell suspension (hereinafter, referred to as cell concentration) has reached a target value. If it is determined that the cell concentration has reached the target value, the process ends. On the other hand, if it is determined that the cell concentration has not reached the target value, the process proceeds to step S7, where it is notified that some abnormality has occurred. If it is determined in step S6 that the cell concentration has not reached the target value, the process may be returned to step S1.
- cell concentration concentration of cells in the concentrated cell suspension
- the cell suspension flowed in from the second flow port 42 is alternately performed with the second liquid transfer in which the cell suspension is allowed to flow out from the first flow port 41.
- the permeated liquid that has permeated through the filtration membrane 20 is discharged from the first discharge port 51 while the first liquid delivery is performed, and the permeation that permeated through the filtration membrane 20 while the second liquid delivery is performed.
- the liquid is discharged from the second outlet 52.
- the permeated liquid that has permeated through the filtration membrane 20 is When the treatment (hereinafter referred to as the first treatment) to be discharged from the discharge port 51 of 1 is performed, an appropriate filtration pressure (a film surface differential pressure of the filtration film 20) on the membrane surface of the filtration membrane 20 is generated.
- region is a diagram showing an example of the distribution (hereinafter, filtered ⁇ that positive region) R A1.
- FIG. 5B is a diagram showing an example of the distribution of filtration pressure at each position on the alternate long and short dash line L1 in FIG. 5A.
- FIG. 5C is a diagram showing an example of the distribution of the liquid transfer speed at each position on the alternate long and short dash line L1 in FIG. 5A.
- the liquid transfer rate refers to the flow rate of the cell suspension flowing on the supply side 31 of the flow path 30.
- the liquid transfer speed on the alternate long and short dash line L1 decreases as it approaches the arrangement position of the first discharge port 51, as shown in FIG. 5C.
- the filtration pressure on the alternate long and short dash line L1 increases as it approaches the arrangement position of the first discharge port 51.
- an area where the filtration pressure becomes higher than the appropriate range an area not hatched in FIG. This is called over-pressure area).
- the filtration pressure excess region relates to the generation of a region in which the flow direction of the cell suspension flowing on the supply side 31 of the flow passage 30 and the flow direction of the permeate flowing on the permeate side 32 of the flow passage 30 are opposite to each other It is thought that it will occur. In the over filtration pressure area, cell clogging is likely to occur, so it is preferable to suppress the occurrence of the over filtration pressure area.
- the first flow port 41 while performing the first liquid transfer in which the cell suspension flowed in from the first flow port 41 is made to flow out from the second flow port 42, the first flow port 41
- the permeated liquid that has permeated through the filtration membrane 20 is discharged from the first discharge port 51 provided at or near the farthest position from the point.
- FIG. 6A shows the second liquid discharge in which the cell suspension flowed in from the second flow port 42 is made to flow out from the first flow port 41, and the permeated liquid that has permeated through the filtration membrane 20 is discharged secondly.
- process of discharging from the outlet 52 (hereinafter referred to as a second process) when doing a diagram showing an example of the distribution of the filtration ⁇ positive region R A2 in the membrane surface of the filtration membrane 20.
- FIG. 6B is a diagram showing an example of the distribution of filtration pressure on the alternate long and short dash line L2 in FIG. 6A.
- FIG. 6C is a diagram showing an example of the distribution of the liquid transfer speed at each position on the alternate long and short dash line L2 in FIG. 6A.
- the liquid transfer speed on the alternate long and short dash line L2 becomes lower as it approaches the arrangement position of the second discharge port 51 as shown in FIG. 6C.
- the filtration pressure on the alternate long and short dash line L2 increases as it approaches the arrangement position of the second discharge port 52, as shown in FIG. 6B.
- a filtration pressure excessive region (a region not hatched in FIG. 6A) is generated in the peripheral region of the corner A1 where the first flow port 41 is disposed in the filtration membrane 20.
- the second flow port 42 The permeated liquid that has permeated through the filtration membrane 20 is discharged from a second discharge port 52 provided at or near the farthest position from the point.
- a second discharge port 52 provided at or near the farthest position from the point.
- FIG. 7A is a diagram showing superimposed filtration ⁇ positive region R A1 and R A2 shown in FIGS. 5A and 6A.
- FIG. 7B is a diagram showing the distributions of the filtration pressure shown in FIG. 5B and FIG. 6B in an overlapping manner, showing an example of the distribution of the filtration pressure on the dashed dotted line L3 in FIG. 7A.
- FIG. 7C is a diagram showing the distribution of the liquid transfer velocity shown in FIG. 5C and FIG. 6C in an overlapping manner, and shows an example of the distribution of the liquid transfer velocity on the alternate long and short dash line L3 in FIG. 7A.
- the entire filtration pressure overrun area in the first process is including the filtration ⁇ positive region R A2 in the processing of, the entire filtering-pressure large region in the second processing, it is difficult to include in the filtration ⁇ positive region R A1 in the first processing.
- FIG. 8 is a diagram showing an example of the configuration of a filtration system 100 according to an embodiment of the disclosed technology that implements a filtration method according to an embodiment of the above-described disclosed technology.
- the filtration system 100 includes a filtration device 1, a first syringe 111, a second syringe 112, a first syringe pump 121, a second syringe pump 122, a first cell concentration detection unit 123, and a second cell concentration detection. It is comprised including the part 124, the collection container 130, the control part 140, the alerting
- Each of the first syringe 111 and the second syringe 112 functions as a container for temporarily storing the cell suspension to be subjected to concentration processing by the filtration device 1 and also includes a means for sending the cell suspension. Act as part of The discharge port of the first syringe 111 is connected to the first flow port 41 of the filtration device 1 through the pipe P1. The discharge port of the second syringe 112 is connected to the second flow port 42 of the filtration device 1 through the pipe P2.
- the first cell concentration detection unit 123 outputs a concentration detection signal D3 indicating the concentration of cells accommodated in the first syringe 111.
- the second cell concentration detection unit 124 outputs a concentration detection signal D4 indicating the concentration of the cells accommodated in the second syringe 112.
- the first syringe pump 121 has a slide mechanism for sliding the plunger of the first syringe 111, and slides the plunger of the first syringe 111 according to the control signal C1 supplied from the control unit 140.
- the liquid suspension of cells contained in the first syringe 111 is sent.
- the first syringe pump 121 outputs a load amount detection signal D1 indicating the size of the load when sliding the plunger of the first syringe 111.
- the load at the time of sliding the plunger of the first syringe 111 increases, for example, as clogging of cells occurs in the filtration membrane 20 and the liquid transfer load increases.
- the second syringe pump 122 has a slide mechanism for sliding the plunger of the second syringe 112, and slides the plunger of the second syringe 112 in response to the control signal C2 supplied from the control unit 140.
- the second syringe pump 122 outputs a load amount detection signal D2 indicating the size of the load when sliding the plunger of the second syringe 112.
- the load at the time of sliding the plunger of the second syringe 112 increases as the clogging of cells occurs in the filtration membrane 20 and the liquid transfer load increases.
- the recovery container 130 is a container from which the permeated liquid that has permeated the filtration membrane 20 is recovered.
- the first discharge port 51 of the filtration device 1 is connected to the recovery container 130 via the pipe P4, and the second discharge port 52 of the filtration device 1 is connected to the recovery container 130 via the pipe P3.
- a valve V1 is provided in the middle of the pipe P1, and a valve V2 is provided in the middle of the pipe P2.
- a valve V3 is provided in the middle of the pipe P3, and a valve V4 is provided in the middle of the pipe P4.
- the valves V1, V2, V3 and V4 each have the form of a solenoid valve, and are controlled to be open or closed according to control signals C3, C4, C5 and C6 supplied from the control unit 140.
- the control unit 140 is configured to include, for example, a microcomputer, and performs liquid transfer control in the filtration system 100 by outputting the control signals C1 to C6 at a predetermined timing. Further, the control unit 140 determines the presence or absence of abnormality in the filtration process based on the load amount detection signals D1 and D2 and the concentration detection signals D3 and D4, and outputs an error signal E1 when it is determined that there is an abnormality.
- the notification unit 150 When the error signal E1 is output from the control unit 140, the notification unit 150 notifies that the filtration system 100 has an abnormality.
- control unit 140 As an initial state, it is assumed that the cell suspension is accommodated in the first syringe 111.
- step S11 the control unit 140 controls the valves V1 and V2 to the open state by outputting the control signals C3 and C4.
- step S12 the control unit 140 drives the first syringe pump 121 by outputting the control signal C1. Further, the control unit 140 controls the valve V4 in the open state and controls the valve V3 in the closed state by outputting the control signals C5 and C6.
- the cell suspension contained in the first syringe 111 flows into the inside of the filtration device 1 from the first flow port 41, and the supply side 31 of the flow path is placed on the membrane surface of the filtration membrane 20. It flows along and flows out of the second flow port 42.
- the cell suspension that has flowed out of the second flow port 42 is accommodated in the second syringe 112.
- the permeated liquid that has permeated through the filtration membrane 20 is discharged from the first discharge port 51 and stored in the collection container 130.
- step S13 the control unit 140 determines whether or not the load amount of the first syringe pump 121 is abnormal based on the load amount detection signal D1.
- the control unit 140 determines that there is an abnormality if the load amount indicated by the load amount detection signal D1 exceeds a predetermined amount, and determines that there is no abnormality otherwise. If the control unit 140 determines that the load amount of the first syringe pump 121 is abnormal, the control unit 140 shifts the process to step S18.
- step S18 the control unit 140 outputs an error signal E1.
- the notification unit 150 notifies that clogging of cells is occurring in the filtration membrane 20 based on the error signal E1.
- step S14 the control unit 140 drives the second syringe pump 122 by outputting the control signal C2. Further, the control unit 140 controls the valve V3 in the open state and controls the valve V4 in the closed state by outputting the control signals C5 and C6.
- the cell suspension contained in the second syringe 112 flows into the inside of the filtration device 1 from the second flow port 42, and the supply side 31 of the flow path is applied to the membrane surface of the filtration membrane 20. It flows along and flows out of the first flow port 41.
- the cell suspension that has flowed out of the first flow port 41 is accommodated in the first syringe 111.
- the permeated liquid that has permeated through the filtration membrane 20 is discharged from the second discharge port 52 and stored in the collection container 130.
- step S15 the control unit 140 determines whether or not the load amount of the second syringe pump 122 is abnormal based on the load amount detection signal D2.
- the control unit 140 determines that there is an abnormality if the load amount indicated by the load amount detection signal D2 exceeds a predetermined amount, and determines that there is no abnormality otherwise. If the control unit 140 determines that the load amount of the second syringe pump 122 is abnormal, the control unit 140 shifts the process to step S18.
- step S18 the control unit 140 outputs an error signal E1.
- the notification unit 150 notifies that clogging of cells is occurring in the filtration membrane 20 based on the error signal E1.
- step S16 the control unit 140 determines whether or not the number of cycles of processing in which a series of processes including each process from step S12 to step S15 is one unit has reached a predetermined number. If the control unit 140 determines that the number of cycles of the series of processes has not reached the predetermined number, the process returns to step S12. On the other hand, when the control unit 140 determines that the number of cycles of the series of processes has reached the predetermined number, the process proceeds to step S17.
- step S17 the control unit 140 performs each of the processes indicated by the concentration detection signal D1 output from the first cell concentration detection unit 123 or the concentration detection signal D2 output from the second cell concentration detection unit 124. It is determined whether the cell concentration after passage has reached the target value. If the control unit 140 determines that the cell concentration after the above processing has not reached the target value, the control unit 140 shifts the processing to step S18. The notification unit 150 notifies that the filter system 100 has some abnormality based on the error signal E1. On the other hand, when the control unit 140 determines that the cell concentration after the above-described processing has reached the target value, the control unit 140 ends the processing. The control unit 140 resets the count value of the number of processing cycles when it is determined that the cell concentration after the above-described processing has not reached the target value, and the processing from step S11 to step S16 is You may carry out again.
- control unit 140 controls the first syringe pump 121 and the second syringe pump 122 to control the delivery of the liquid passing through the first flow port 41 and the second flow port 42.
- the first liquid transfer control is performed.
- control unit 140 performs opening / closing control of the valves V3 and V4 in conjunction with the first liquid transfer control, so that the permeated liquid discharged from the first discharge port 51 and the second discharge port 52 A second liquid transfer control is performed to control the liquid transfer.
- control unit 140 causes the cell suspension that has flowed in from the first flow port 41 to be discharged from the second flow port 42 as the first liquid transfer control, and The cell suspension that has flowed in from the second flow port 42 is alternately carried out with the second liquid transfer that is discharged from the first flow port 41.
- the control unit 140 discharges the permeated liquid that has permeated the filtration membrane 20 from the first discharge port 51, and performs the second liquid transfer. During this time, the permeated liquid that has permeated through the filtration membrane 20 is discharged from the second discharge port 52.
- Figure 10 is a diagram showing an example of the distribution of the filtration ⁇ positive region R A3 in the case of carrying out the filtration method according to the first comparative example using the filtration device 1X according to the comparative example, the film surface of the filtration membrane 20 is there.
- the filtration device 1X has only one discharge port for discharging the permeated liquid that has permeated through the filtration membrane 20.
- the discharge port 50 is an end of the unit flow path 30a to which the first flow port 41 is connected opposite to the end on the side where the first flow port 41 is provided. It is provided on the transmission side of the unit.
- the process of discharging the permeated liquid from the discharge port 50 is performed while the cell suspension flowed in from the first flow port 41 is made to flow out from the second flow port 42. Do it continuously. That is, in the filtration method according to the first comparative example, switching of the liquid transfer direction on the supply side is not performed.
- the area of the filtration pressure appropriate area R A3 is the filtration method according to the embodiment of the disclosed technology. It becomes smaller than the area of filtration pressure appropriate area R A1 (see FIG. 5) when applied.
- the filtration pressure appropriate region R A3 does not cover the entire filtration membrane 20. Therefore, according to the filtration method according to the first comparative example, the risk of clogging of cells is increased and the cell recovery rate is reduced, as compared with the filtration method according to the embodiment of the disclosed technology.
- FIG 11 is a diagram showing a second case where the filtration method was performed according to the comparative example, an example of the distribution of filtered ⁇ positive region R A3 and R A4 in the membrane surface of the filtration membrane 20 with a filtering device 1X .
- a filtration membrane is performed while performing a first treatment of discharging the permeated liquid from the discharge port 50 and a second liquid transfer in which the cell suspension flowed in from the second flow port 42 is made to flow out of the first flow port 41.
- 20 and the second treatment of discharging the permeated liquid that has permeated 20 from the discharge port 50 alternately.
- a portion of the filtered-pressure large region in the first processing can be included in the filtering ⁇ positive region R A4 in the second processing, the filtration in the second processing some of-pressure large area, can be included in the filtering ⁇ positive region R A3 in the first processing.
- the discharge port for discharging the permeate is one, the entire filtering-pressure large region in the first processing, it can not be included in the filtering ⁇ positive region R A4 in the second processing.
- the entire filtration pressure discrepancy large region in the second processing can not be included in the filtering ⁇ positive region R A3 in the first processing.
- the filtration ⁇ positive region R A3 in the first processing, a region combining the filtered ⁇ positive region R A4 in the second process, does not cover the entire filtration membrane 20. Therefore, according to the filtration method according to the second comparative example, the risk of clogging of cells is increased and the cell recovery rate is decreased, as compared with the filtration method according to the embodiment of the disclosed technology.
- Figure 12 is a view showing a case embodying the filtration method according to the third comparative example using a filtration device 1Y according to the comparative example, an example of a distribution of filtering ⁇ positive region R A5 in the membrane surface of the filtration membrane 20 is there.
- the filtering device 1Y has only one outlet for discharging the permeated liquid that has permeated through the filtration membrane 20.
- the discharge port 50 is a unit positioned midway between the unit flow path 30a provided with the first flow port 41 and the unit flow path 30a to which the second flow port 42 is connected.
- the passage 30 a is provided on the transmission side of the end on the opposite side to the end on the side where the first communication port 41 and the second communication port 42 are provided.
- the process of discharging the permeated liquid from the discharge port 50 is performed while the cell suspension flowed in from the first flow port 41 is made to flow out from the second flow port 42. Do it continuously. That is, in the filtration method according to the third comparative example, switching of the liquid transfer direction on the supply side is not performed.
- filtration when the area of the filtration ⁇ positive region R A5 was carried filtration method according to the first comparative example using the filtration device 1X It becomes larger than the area of pressure appropriate area R A3 (see FIG. 5).
- filtered ⁇ area of the positive region R A5 is filtering in the case of carrying out the filtration method according to an embodiment of the disclosed technique ⁇ positive region R A1 (see FIG. 5)
- filtration ⁇ positive region R A5 does not cover the entire filtration membrane 20. Therefore, according to the filtration method according to the third comparative example, the risk of clogging of cells is increased and the cell recovery rate is decreased, as compared with the filtration method according to the embodiment of the disclosed technology.
- Figure 13 is a diagram showing a fourth case of carrying out the filtration method according to the comparative example, an example of the distribution of filtered ⁇ positive region R A5 and R A6 in the membrane surface of the filtration membrane 20 with filtration devices 1Y .
- a filtration membrane is performed while performing a first treatment of discharging the permeated liquid from the discharge port 50 and a second liquid transfer in which the cell suspension flowed in from the second flow port 42 is made to flow out of the first flow port 41.
- 20 and the second treatment of discharging the permeated liquid that has permeated 20 from the discharge port 50 alternately.
- a portion of the filtered-pressure large region in the first processing can be included in the filtering ⁇ positive region R A6 in the second processing, the filtration in the second processing -pressure part of a large area, can be included in the filtering ⁇ positive region R A5 in the first processing.
- the entire filtration overpressure region in the first treatment is a filtration pressure appropriate region in the second treatment. It can not be included in R A6 .
- the entire filtration pressure discrepancy large region in the second processing can not be included in the filtering ⁇ positive region R A5 in the first processing.
- the filtered ⁇ positive region R A5 in the first processing, a region combining the filtered ⁇ positive region R A6 in the second process, does not cover the entire filtration membrane 20. Therefore, according to the filtration method according to the fourth comparative example, the risk of clogging of cells is increased and the cell recovery rate is decreased, as compared with the filtration method according to the embodiment of the disclosed technology.
- the first liquid delivery in which the cell suspension flowed in from the first flow port 41 is made to flow out from the second flow port 42 is used.
- the process is alternately performed with the second process of discharging the liquid.
- the entire filtering-pressure large region in the first processing including the filtering ⁇ positive region R A2 in the second processing, the entire filtering-pressure large region in the second processing, the filtration pressure in the first processing It can be included in the appropriate area RA1 . That is, the filtration ⁇ positive region R A1 in the first processing, a region combining the filtered ⁇ positive region R A2 in the second process, covers the entire filtration membrane 20.
- the time average value of the filtration pressure on the membrane surface of the filtration membrane 20 can be made substantially uniform. Therefore, in comparison with the first to fourth comparative examples described above, the risk of cell clogging can be suppressed, and the cell recovery rate can be enhanced.
- FIG. 14A shows the state of the filtration membrane after the membrane separation treatment of a liquid containing simulated particles simulating cells by the filtration method (see FIG. 11) according to the second comparative example using the filtration apparatus 1X.
- FIG. FIG. 14B is a view showing the state of the filtration membrane after the membrane separation treatment of the liquid containing the simulated particles simulating the cells by the filtration method according to the embodiment of the disclosed technology using the filtration device 1.
- the colored part is the part where the simulated particles are deposited, and the part where the coloring density is high is the part where the filtration pressure is relatively high.
- the portion with high color density and the portion with low color density are clearly separated. This indicates that the difference between the high and low parts of the filtration pressure on the membrane surface of the filtration membrane is remarkable, that is, the filtration pressure on the membrane surface of the filtration membrane is nonuniform.
- the coloring density is substantially uniform as a whole. This indicates that the filtration pressure on the membrane surface of the filtration membrane is substantially uniform.
- the area of the filtration membrane is increased while securing the uniformity of the filtration pressure on the membrane surface of the filtration membrane 20 It is possible to Therefore, the filtration efficiency can be enhanced while suppressing the decrease in the cell recovery rate.
- FIG. 15 is a plan view showing the configuration of a filtration device 1A according to a second embodiment of the disclosed technology.
- the filtration apparatus 1A according to the first embodiment is that the flow path 30 includes a plurality of unit flow paths 30a and a plurality of unit flow paths 30c whose flow path length is shorter than the unit flow path 30a. It differs from the filtration device 1.
- the flow path 30 has a portion in which the distance from one of the folding portions 30b to the next folding portion 30b is short as compared to the other portions.
- the length of the unit flow path 30 a is La
- the length of the unit flow path 30 c is Lc
- the first liquid transfer is performed to cause the cell suspension flowed from the first flow port 41 to flow out from the second flow port 42.
- the first treatment of discharging the permeated liquid from the first discharge port 51 and the second liquid transfer of causing the cell suspension flowed in from the second flow port 42 to flow out of the first flow port 41 While performing the second treatment of discharging the permeated liquid from the second outlet 52 is alternately performed, as shown in FIG.
- the filtering ⁇ positive region R A2 can not be included in the filtering ⁇ positive region R A2 in the process, if the entire filtration pressure discrepancy large region generated in the second process can not be included in the filtering ⁇ positive region R A1 in the first process, the first In any of the treatment and the second treatment, filtration pressure In the portion where the region, the higher the risk of clogging of the cells occurs, there is a risk that the recovery rate of the cells is reduced.
- the flow passage portion which becomes the filtration pressure excessive region is eliminated in any of the first process and the second process. Therefore, the risk of clogging of cells can be suppressed.
- one end and the other end of the flow path 30 are disposed on the same side of the filtration membrane 20 having a rectangular shape, and accordingly, the first flow port 41 and the second flow port 42
- the present invention is not limited to this embodiment.
- one end of the flow path 30 and the first flow port 41 are disposed in the vicinity of the corner A1 of the filtration membrane 20 having a rectangular shape
- the other end of the flow path 30 and the second The flow port 42 may be disposed in the vicinity of the corner A3 which is a diagonal of the corner A1 of the filtration membrane 20.
- the first discharge port 51 is disposed, for example, in the vicinity of the corner A3 of the filtration membrane 20, that is, immediately below the second flow port 42, and the second discharge port 52 is, for example, the filtration membrane 20. It is disposed in the vicinity of the corner A1 of the second embodiment, that is, immediately below the first flow port 41.
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Abstract
Description
図1は、開示の技術の実施形態に係る濾過装置1の構成の一例を示す平面図、図2A、図2B、図2C及び図2Dは、それぞれ、図1における2A-2A線、2B-2B線、2C-2C線及び2D-2D線に沿った断面図である。
0.5W1≦d1≦W1 ・・・(1)
0.5W2≦d2≦W2 ・・・(2)
濾過膜20において細胞の目詰まりの発生を抑制する観点から、下記の(3)式及び(4)式を満たすことがより好ましい。
0.7W1≦d1≦W1 ・・・(3)
0.7W2≦d2≦W2 ・・・(4)
なお、図4のフローチャートでは大まかな流れを記載したが、ステップS2はステップS1の第1の送液中に繰り返し行われる。そして第1の送液時に異常がなく第1の送液が終了する条件が成立すると、ステップS3に移行する。第1の送液が終了する条件は、例えば所定の処理量をすべて処理し終わった場合である。また、ステップS2の判定を、第1の送液が終了する条件が成立した後に行ってもよい。
なお、図4のフローチャートでは大まかな流れを記載したが、ステップS4はステップS3の第2の送液中に繰り返し行われる。そして第2の送液時に異常がなく第2の送液が終了する条件が成立すると、ステップS5に移行する。第2の送液が終了する条件は、例えば所定の処理量をすべて処理し終わった場合である。また、ステップS4の判定を、第2の送液が終了する条件が成立した後に行ってもよい。
図15は、開示の技術の第2の実施形態に係る濾過装置1Aの構成を示す平面図である。濾過装置1Aは、流路30が、複数の単位流路30aと、流路長が単位流路30aよりも短い複数の単位流路30cとを含んでいる点が、第1の実施例に係る濾過装置1と異なる。換言すれば、濾過装置1Aにおいて、流路30は、折り返し部30bのうちの1つから次の折り返し部30bまでの距離が、他の部分と比較して短い部分を有する。ここで、単位流路30aの長さをLa、単位流路30cの長さをLcとした場合、下記の(5)式を満たすことが好ましい。
0.5×La<Lc<La ・・・(5)
Claims (10)
- 蛇行した流路と、
前記流路の供給側と透過側とを隔てる濾過膜と、
前記流路の前記供給側の一端に設けられた第1の流通口と、
前記流路の前記供給側の他端に設けられた第2の流通口と、
前記流路の前記透過側に設けられた第1の排出口と、
前記流路の前記透過側の前記第1の排出口とは異なる位置に設けられた第2の排出口と、
を含む濾過装置。 - 第1の流通口の中心から、前記濾過膜の膜面の最遠位置までの距離をW1とし、前記第1の流通口の中心から前記第1の排出口の中心までの距離をd1とし、
第2の流通口の中心から、前記濾過膜の膜面の最遠位置までの距離をW2とし、前記第2の流通口の中心から前記第2の排出口の中心までの距離をd2としたとき、
0.5W1≦d1≦W1を満たし、
0.5W2≦d2≦W2を満たす
請求項1に記載の濾過装置。 - 前記流路は複数の折り返し部を有し、前記折り返し部のうちの1つから次の折り返し部までの各距離が互いに同じである
請求項1または請求項2に記載の濾過装置。 - 前記流路の一端及び他端並びに一方の側の前記折り返し部の各々が、第1の直線の上に設けられ、他方の側の前記折り返し部の各々が前記第1の直線と平行な第2の直線の上に設けられている
請求項3に記載の濾過装置。 - 前記流路は複数の折り返し部を有し、前記折り返し部のうちの1つから次の折り返し部までの距離が他の部分と比較して短い部分を有する
請求項1または請求項2に記載の濾過装置。 - 請求項1から請求項5のいずれか1項に記載の濾過装置と、
前記第1の流通口及び前記第2の流通口を通過する液体の送液を制御する第1の送液制御を行い、前記第1の送液制御と連動して、前記第1の排出口及び前記第2の排出口から排出される液体の送液を制御する第2の送液制御を行う制御部と、
を含む濾過システム。 - 前記制御部は、
前記第1の送液制御として、前記第1の流通口から流入させた液体を、前記第2の流通口から排出させる第1の送液と、前記第2の流通口から流入させた液体を、前記第1の流通口から排出させる第2の送液とを交互に行い、
前記第2の送液制御として、前記第1の送液を行っている間、前記濾過膜を透過した液体を前記第1の排出口から排出し、前記第2の送液を行っている間、前記濾過膜を透過した液体を前記第2の排出口から排出する
請求項6に記載の濾過システム。 - 前記液体は細胞懸濁液である
請求項6または請求項7に記載の濾過システム。 - 請求項1から請求項5のいずれか1項に記載の濾過装置を用いた液体の濾過方法であって、
前記第1の流通口から流入させた液体を前記第2の流通口から流出させる第1の送液と、前記第2の流通口から流入させた液体を前記第1の流通口から流出させる第2の送液とを交互に行い、
前記第1の送液を行っている間、前記濾過膜を透過した液体を前記第1の排出口から排出し、前記第2の送液を行っている間、前記濾過膜を透過した液体を前記第2の排出口から排出する
濾過方法。 - 細胞懸濁液を前記第1の送液及び前記第2の送液の対象とする
請求項9に記載の濾過方法。
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- 2018-09-19 CN CN201880061083.1A patent/CN111148564A/zh active Pending
- 2018-09-19 EP EP18857812.4A patent/EP3673979A4/en not_active Withdrawn
- 2018-09-19 JP JP2019543681A patent/JPWO2019059241A1/ja not_active Abandoned
- 2018-09-19 KR KR1020207008071A patent/KR20200041968A/ko active IP Right Grant
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Also Published As
Publication number | Publication date |
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EP3673979A1 (en) | 2020-07-01 |
CN111148564A (zh) | 2020-05-12 |
KR20200041968A (ko) | 2020-04-22 |
JPWO2019059241A1 (ja) | 2020-10-15 |
EP3673979A4 (en) | 2020-10-21 |
US20200215491A1 (en) | 2020-07-09 |
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