Concentration device, analytical system comprising the concentration device and concentration method

Abstract

A concentration device, an analysis system including the concentration device, and a concentration method are provided. The concentration device of the present application includes a frame, a separation membrane that divides an internal space of the frame in a manner that forms a flow path inside the frame, a first supply portion that supplies a first liquid from a first position of the frame to the flow path in a manner that the first liquid flows along the separation membrane in a first direction, a second supply portion that supplies a second liquid from a second position of the frame to the flow path in a manner that the second liquid flows along the separation membrane in a second direction opposite to the first direction, and a third supply portion that supplies a third liquid containing a target component having a size that does not pass through the separation membrane from a third position of the frame to the flow path, the third position being located between the first position and the second position in the first direction.

Description

TECHNICAL FIELD

[0001] The present application relates to a concentration device, an analysis system including the concentration device, and a concentration method. BACKGROUND

[0002] Tangential-flow-filtration (TFF) is sometimes used in concentration or desalination of viruses, proteins, peptides, nucleic acids, or the like. For example, a method for producing viruses described in Japanese Patent No. 5548207 includes concentrating a stabilized virus recovery product by tangential-flow-filtration.

[0003] In a concentration device using tangential-flow-filtration, a solution containing a component to be concentrated (hereinafter, referred to as a target component) flows along a separation membrane. The pore diameter of the separation membrane is smaller than the target component. Thus, a component smaller than the pore diameter of the separation membrane permeates the separation membrane, and the target component does not permeate the separation membrane. The solution containing the target component is recovered into a storage tank and flows along the separation membrane again. Thus, by circulating the solution containing the target component in a flow path including the separation membrane and the storage tank, the target component in the solution is concentrated. SUMMARY

[0004] However, in the conventional concentration device using tangential-flow-filtration, in order to concentrate the target component in the solution, the circulation of the solution needs to be repeated, and thus a large amount of time is required. In addition, the amount of the entire solution containing the target component after concentration becomes large. Thus, the processing of the solution containing a certain amount of the target component becomes complicated. Therefore, it is desirable to recover a small amount of solution containing a certain amount of the target component.

[0005] An object of the present application is to provide a concentration device, an analysis system including the concentration device, and a concentration method, which can concentrate a target component in a liquid in a short time and can recover a small amount of liquid containing the target component after concentration.

[0006] A concentration device according to an aspect of the present application includes a frame, a separation membrane that divides an internal space of the frame in a manner to form a flow path inside the frame, a first supply portion that supplies a first liquid to the flow path from a first position of the frame in a manner that the first liquid flows along the separation membrane in a first direction, a second supply portion that supplies a second liquid to the flow path from a second position of the frame in a manner that the second liquid flows along the separation membrane in a second direction opposite to the first direction, and a third supply portion that supplies a third liquid containing a target component having a size that does not permeate the separation membrane to the flow path from a third position of the frame, the third position being located between the first position and the second position in the first direction.

[0007] The concentration method according to another aspect of the present application includes a step of supplying a first liquid from a first position of a frame to a flow path along a separation membrane in the frame in a manner that the first liquid flows along the separation membrane in a first direction along the flow path, a step of supplying a second liquid from a second position of the frame to the flow path in a manner that the second liquid flows along the separation membrane in a second direction opposite to the first direction, and a step of supplying a third liquid containing a target component having a size that does not pass through the separation membrane from a third position of the frame to the flow path, the third position being located between the first position and the second position in the first direction. BRIEF DESCRIPTION OF DRAWINGS

[0008] Figure 1 is a schematic view showing a structure of an analysis system including a concentration device of one embodiment.

[0009] Figure 2 is a graph showing an example of a chromatogram generated on the basis of an output signal of a detector of a liquid chromatograph.

[0010] Figure 3 is a timing chart showing a transition of a state of each portion in the analysis system of Figure 1 .

[0011] Figure 4 is a schematic view showing a concentration operation of the analysis system of Figure 1 .

[0012] Figure 5 is a schematic view showing a recovery operation of the analysis system of Figure 1 .

[0013] Figure 6 is a graph showing chromatograms obtained in Example 1 and Comparative Example 1.

[0014] Figure 7 is a graph showing a chromatogram obtained in Example 2. DETAILED DESCRIPTION

[0015] Hereinafter, a concentration device, an analysis system, and a concentration method of one embodiment will be described in detail with reference to the drawings.

[0016] (1) Structure of Analysis System

[0017] Figure 1 is a schematic view showing a structure of an analysis system including a concentration device of one embodiment. The analysis system 100 includes a liquid chromatograph 1 and a concentration device 10. The liquid chromatograph 1 includes a pump 3, an autosampler 4, a separation column 5, a detector 6, and pipes p11 to p15.

[0018] In the present embodiment, the ejection port of the pump 3 is connected to the autosampler 4 through the pipe p11. The autosampler 4 is connected to the introduction port of the separation column 5 through the pipe p12. The lead-out port of the separation column 5 is connected to the introduction port of the detector 6 through the pipe p13.

[0019] A switching valve V3 is provided at the lead-out port of the detector 6. The switching valve V3 has one liquid inlet, a first liquid outlet, and a second liquid outlet. The lead-out port of the detector 6 is connected to the liquid inlet of the switching valve V3 through the pipe p14. The pipe p15 is connected to the first liquid outlet of the switching valve V3. The state in which the liquid inlet of the switching valve V3 is connected to the first liquid outlet is referred to as a first state. The state in which the liquid inlet of the switching valve V3 is connected to the second liquid outlet is referred to as a second state.

[0020] An eluent, which is a mobile phase, is housed in the eluent container 2. The pump 3 sucks the eluent from the eluent container 2 and guides the sucked eluent to the separation column 5 through the pipe p11, the autosampler 4, and the pipe p12.

[0021] The autosampler 4 introduces a sample, which is an analysis target, into the eluent guided from the pump 3 to the separation column 5. Thereby, the eluent containing the sample is introduced to the separation column 5. The separation column 5 separates the sample in the eluent in terms of components in time. The eluent containing the separated sample is introduced from the lead-out port of the separation column 5 to the introduction port of the detector 6 through the pipe p13. The detector 6 detects the components of the separated sample. The detector 6 is, for example, an ultraviolet (UV) detector. A chromatogram is generated based on the output signal of the detector 6. Hereinafter, these actions are referred to as analysis actions. After that, the eluent containing the sample led out from the detector 6 is guided to the switching valve V3 through the pipe p14. When the switching valve V3 is switched to the first state, the eluent guided to the switching valve V3 is guided to a waste tank or the like, not shown, through the pipe p15.

[0022] In the present embodiment, the concentration device 10 is used to concentrate one or more components in the eluent led out from the detector 6 of the liquid chromatograph 1. The concentration device 10 includes a frame 20, a control section 30, a pump 8a, a pump 8b, a fraction collector F1, a flow rate adjustment valve V1, a switching valve V3, a switching valve V4, an on-off valve V5, and pipes p1 to p5. The frame 20 is formed in a cylindrical shape having an internal space extending in a direction Y, and one end portion and the other end portion are closed. The cross-sectional shape of the frame 20 can be circular, elliptical, polygonal, or other shapes.

[0023] Inside the frame 20, a separation membrane ME is arranged parallel to the Y direction, dividing the space into a flow path FP and a discharge space SP. In this embodiment, an ultrafiltration membrane can be used as the separation membrane ME. The ultrafiltration membrane has a pore size of approximately 0.001 μm to 0.01 μm. Here, a separation membrane with a pore size smaller than the size of the target component to be concentrated can be used as the separation membrane ME. In this embodiment, a porous sintered body 21 is provided in the discharge space SP to support the separation membrane ME. Alternatively, a mesh-like sieve can be provided instead of the porous sintered body 21.

[0024] The frame 20 of the concentration device 10 has a first supply port P1 to a third supply port P3, a first discharge port P4, and a second discharge port P5. The first supply port P1 to the third supply port P3 and the first discharge port P4 are configured to connect the flow path FP inside the frame 20 with the outside of the frame 20.

[0025] In this embodiment, the first supply port P1 is located at one end of the flow path FP of the frame 20, and the first discharge port P4 is located at the other end of the flow path FP of the frame 20. In this embodiment, the first supply port P1 and the first discharge port P4 are located on the same straight line.

[0026] Hereinafter, the direction from the first supply port P1 towards the first discharge port P4 is referred to as the first direction a, and the direction from the first discharge port P4 towards the first supply port P1 is referred to as the second direction b. The first direction a and the second direction b face each other. In this embodiment, the first direction a and the second direction b are parallel to the direction Y. Furthermore, the first supply port P1 and the first discharge port P4 may not be on the same straight line, and the first direction a and the second direction b may not be on a straight line. That is, the first direction a and the second direction b may also form other angles close to 180°.

[0027] The second supply port P2 is located in the Y direction between the first supply port P1 and the first discharge port P4. Furthermore, the third supply port P3 is located in the Y direction between the first supply port P1 and the second supply port P2. In this embodiment, the third supply port P3 is located closer to the first supply port P1 in the Y direction than the second supply port P2. The second discharge port P5 is configured to communicate with the discharge space SP within the frame 20 and the outside of the frame 20.

[0028] The first supply port P1 of the frame 20 is connected to the nozzle of the pump 8a via pipe p1 and flow regulating valve V1. Pump 8a draws in the eluent contained in the eluent container 7a and guides it to the first supply port P1 via flow regulating valve V1 and pipe p1. Thus, the eluent is supplied from the first supply port P1 into the flow path FP of the frame 20, and the eluent supplied from the first supply port P1 flows along the separation membrane ME in the first direction a. The flow regulating valve V1 is configured to switch the flow rate of the eluent discharged to pipe p1 to a first flow rate and a second flow rate greater than the first flow rate. Hereinafter, the state in which the flow regulating valve V1 discharges the first flow rate of eluent to pipe p1 is referred to as the first state, and the state in which the second flow rate of eluent is discharged is referred to as the second state.

[0029] The second supply port P2 of the frame 20 is connected to the nozzle of the pump 8b via pipe p2. The pump 8b draws in the eluent contained in the eluent container 7b and guides it to the second supply port P2 via pipe p2. Thus, the eluent is supplied from the second supply port P2 into the flow path FP of the frame 20, flowing along the separation membrane ME in the second direction b and also in the first direction a. In this embodiment, the eluent contained in the eluent containers 7a and 7b is the same as the eluent contained in the eluent container 2.

[0030] The third supply port P3 of the housing 20 is connected to the second liquid outlet of the switching valve V3 via pipe p3. When the switching valve V3 is switched to the second state, the eluent discharged from the detector 6 is guided to the third supply port P3 of the housing 20 via pipe p3. Thus, the eluent containing the target component is supplied from the third supply port P3 toward the separation membrane ME in the flow path FP of the housing 20.

[0031] The switching valve V4 has a liquid inlet, a first liquid outlet, and a second liquid outlet. The first discharge port P4 of the frame 20 is connected to the liquid inlet of the switching valve V4 via pipe p4. The first liquid outlet of the switching valve V4 is connected to the fraction collector (FRC) F1 via pipe p16. The eluent from the flow path FP of the frame 20 is discharged from the first discharge port P4 through pipe p4 to the liquid inlet of the switching valve V4. When the switching valve V4 is switched to the first state, the eluent from the flow path FP of the frame 20 is guided to the fraction collector F1 via pipe p16. When the switching valve V4 is switched to the second state, the eluent from the flow path FP of the frame 20 is guided to a waste liquid tank (not shown) via pipe p17.

[0032] One or more sample collection containers SC are configured in the fraction collector F1. The fraction collector F1 recovers a certain amount of eluent containing the target component guided from the flow path FP of the frame 20 into the sample collection container SC.

[0033] Among the various components in the eluent flowing along the separation membrane ME within the flow path FP of the housing 20, components with a size smaller than the pore size of the separation membrane ME permeate through the separation membrane ME along with the eluent. The eluent and components that have permeated through the separation membrane ME from the flow path FP are guided to the discharge space SP of the housing 20. The second discharge port P5 of the housing 20 is connected to the on / off valve V5 via pipe p5. When the on / off valve V5 is in the open state, the eluent in the discharge space SP is discharged through pipe p5 to a waste liquid tank (not shown) or the like.

[0034] The control unit 30 controls the operation of the flow control valve V1, switching valve V3, switching valve V4, on / off valve V5, pump 8a, pump 8b, and fraction collector F1 of the pump 3, automatic sampler 4, detector 6, and concentration unit 10. Thus, in this embodiment, the control unit 30 controls the components of the liquid chromatograph 1 and the concentration unit 10. Furthermore, the control unit controlling the liquid chromatograph 1 and the control unit controlling the concentration unit 10 can also be provided separately.

[0035] The control unit 30 includes a central processing unit (CPU), a read-only memory (ROM), and a random access memory (RAM). The ROM stores in advance an analysis program for performing the analysis operations of the liquid chromatograph 1, a concentration program for causing the concentration device 10 to perform the concentration operations described later, and the dissolution time of the target component. The CPU performs the analysis and concentration operations by executing the analysis program and concentration program stored in the ROM on the RAM.

[0036] Figure 2 This is a graph representing an example of a chromatogram generated based on the output signal of detector 6 of liquid chromatograph 1. The horizontal axis of the chromatogram represents dissolution time (holding time), and the vertical axis represents signal intensity. Figure 2 The chromatogram shown contains multiple peaks A to C representing various components in the sample that were separated over time by separation column 5.

[0037] (2) Analyze the actions of system 100

[0038] The analytical system 100 of this embodiment performs analytical operations based on the liquid chromatograph 1 and concentration operations based on the concentration device 10. The concentration operation includes aggregation and recovery operations. The aggregation operation is the operation of collecting the target component in the eluent to a predetermined position on the separation membrane ME. The recovery operation is the operation of discharging the target component collected by the aggregation operation together with the eluent from the housing 20 and recovering it into the sample receiving container SC.

[0039] Figure 3 It means Figure 1 A time-series diagram showing the state transitions of various components in liquid chromatograph 1. Figure 4 This is a schematic diagram representing the aggregation action in the condensation action of the analysis system 100. Figure 5 This is a schematic diagram representing the recovery action in the concentration process of the analysis system 100. Figure 4 and Figure 5 In the diagram, the flow of liquid is represented by thick lines. Here, for... Figure 2 The composition corresponding to peak C in the chromatogram is explained as the concentration of the target component.

[0040] Figure 3 The diagram shows the time-varying states of flow control valve V1, switching valve V3, switching valve V4, on / off valve V5, and pumps 8a and 8b during the analysis and concentration operations. Figure 3 In the example, the analysis action begins at time t1, the condensation action is performed from time t2 to time t5, the aggregation action within the condensation action is performed from time t3 to time t4, and the retrieval action is performed from time t4 to time t5.

[0041] first, Figure 4 The control unit 30 switches the switching valve V3 to the first state. As a result, the eluent ejected from the pump 3 is guided to the detector 6 through piping p12, the automatic sampler 4, piping p13, and the separation column 5. At this time, the sample is introduced into the eluent via the automatic sampler 4. The sample components in the eluent are then separated by the separation column 5. In this state, the separated components are detected by the detector 6. The control unit 30 generates, for example, a generator based on the output signal of the detector 6. Figure 2 The chromatography is performed. The eluent from detector 6 is discharged into the waste tank, etc., via switching valve V3 and piping p15. At time t1, pumps 8a and 8b are closed, switching valve V4 is in the second state, and on / off valve V5 is closed.

[0042] Next, at a predetermined time t2 from time t1, the control unit 30 turns on pump 8a and switches the flow regulating valve V1 to the first state. As a result, the eluent in the eluent container 7a is supplied to the flow path FP within the frame 20 via pump 8a, piping p1, and the first supply port P1 of the frame 20. The eluent supplied to the flow path FP via the first supply port P1 flows along the separation membrane ME in the first direction a. Meanwhile, the control unit 30 turns on pump 8b. As a result, the eluent in the eluent container 7b is supplied to the flow path FP within the frame 20 via pump 8b, piping p2, and the second supply port P2 of the frame 20. The eluent supplied to the flow path FP from the second supply port P2 flows along the separation membrane ME in both the first direction a and the second direction b. In this case, the eluent flowing from the first supply port P1 in the first direction a and the eluent flowing from the second supply port P2 in the second direction b collide between the first supply port P1 and the second supply port P2. Additionally, the control unit 30 opens the on / off valve V5, thereby allowing discharge from the discharge space SP within the frame 20.

[0043] Detected using detector 6 and Figure 2 The aggregation process begins at time t3, starting from the component corresponding to peak C (hereinafter referred to as target component c). At time t3, the control unit 30 switches the switching valve V3 to the second state. As a result, the eluent containing target component c, exported from the detector 6, is supplied to the flow path FP within the housing 20 through the switching valve V3, the piping p3, and the third supply port P3 of the housing 20.

[0044] The eluent containing the target component c is supplied to the collision point of the eluent supplied from the first supply port P1 and flowing in the first direction a and the eluent supplied from the second supply port P2 and flowing in the second direction b.

[0045] exist Figure 4 In this example, the third supply port P3 is positioned closer to the first supply port P1 than the second supply port P2. In this case, the flow rate of the eluent in pipe P2 is set to be greater than the flow rate of the eluent in pipe P1, so that the eluent from the third supply port P3 is supplied to the point where the eluent supplied from the first supply port P1 and the eluent supplied from the second supply port P2 collide.

[0046] Thus, since the eluent containing the target component c is supplied from the third supply port P3 to the collision point of the eluent flowing in the first direction a and the eluent flowing in the second direction b, the target component c accumulates on the separation membrane ME. Furthermore, during the accumulation of the target component c, a portion of the eluent flowing along the separation membrane ME in the first direction a and a portion of the eluent flowing in the second direction b permeate through the separation membrane ME via tangential flow filtration. Furthermore, components in the eluent supplied from the third supply port P3 that have a size smaller than the pore size of the separation membrane ME permeate through the separation membrane ME. The eluent and components that have permeated through the separation membrane ME are guided from the second discharge port P5 through piping p5 and on / off valve V5 to a waste liquid tank (not shown).

[0047] The aggregation of target component c is considered complete when a predetermined time has elapsed since the start of the aggregation operation at time t3, or when the signal strength of detector 6 falls below a predetermined value. The recovery operation then begins at time t4. At time t4, control unit 30 switches switching valve V3 to the first state. This stops the supply of eluent from the third supply port P3 to the flow path FP of the housing 20. Additionally, control unit 30 shuts down pump 8b. This stops the supply of eluent from the second supply port P2 to the flow path FP of the housing 20. Furthermore, control unit 30 switches switching valve V4 to the first state. Additionally, control unit 30 closes the on / off valve V5.

[0048] In this state, the control unit 30 switches the flow regulating valve V1 to the second state. As a result, eluent continues to be supplied to the first supply port P1. In this case, the amount of eluent supplied to the first supply port P1 during the recovery operation is greater than the amount supplied during the aggregation operation. Consequently, a flow of eluent in the first direction a from the first supply port P1 toward the first discharge port P4 is formed within the flow path FP of the frame 20. As a result, the eluent within the flow path FP of the frame 20 and the target component c aggregated on the separation membrane ME are guided from the first discharge port P4 through pipe p4, switching valve V4, and pipe p16 to the fraction collector F1. Here, the eluent containing the target component c is recovered into the sample collection container SC disposed in the fraction collector F1. The concentration operation performed by the concentration device 10 ends at the moment t5 when the eluent containing the target component c is recovered into the sample collection container SC.

[0049] (3) Example 1 and Comparative Example 1

[0050] In Example 1, the recovery rate of the target component during the aggregation operation of the concentration device 10 was evaluated. In this Example 1, the flow rates of the first supply port P1, second supply port P2, third supply port P3, first discharge port P4, and second discharge port P5 of the concentration device 10 were controlled using a pump, syringe, switching valve, and mass flow meter. An ultrafiltration membrane with a molecular weight cutoff of 10 kDa was used as the separation membrane ME. A UV (ultraviolet) detector with a wavelength of 280 nm was connected to the first discharge port P4. Bovine serum albumin (BSA), manufactured by Sigma-Aldrich, was used as the target component. A BSA solution with a concentration of 0.1 mg / mL was prepared by dissolving 1.0 mg of BSA in 10 mL of ultrapure water.

[0051] During the aggregation process, a 0.1 mg / mL BSA solution was injected into the flow path FP through the third supply port P3 using a 1 mL manual injector. Ultrapure water was supplied into the flow path FP through the first supply port P1 and the second supply port P2. The flow rate at the first supply port P1 was set to 0.5 mL / min, and the flow rate at the second supply port P2 was set to 1.7 mL. Additionally, the flow rate at the first discharge port P4 and the second discharge port P5 was set to 1.2 mL / min. As a result, the BSA solution aggregated on the separation membrane ME for 2.5 minutes.

[0052] During the recovery process, the flow rate at the first supply port P1 is set to 1.2 mL / min, and the flow rates at the second supply ports P2 and the third supply port P3 are set to 0 mL / min. Additionally, the BSA accumulated on the separation membrane ME is discharged from the first discharge port P4 over approximately 1 minute. At this time, the flow rate at the second discharge port P5 is set to 0 mL / min, and the tangential flow filtration flow rate is also set to 0 mL / min. The BSA solution discharged from the first discharge port P4 is directed to the UV detector.

[0053] In Comparative Example 1, without aggregation, a BSA solution with a concentration of 0.1 mg / mL was injected into the flow path FP through the third supply port P3 using a 1 mL manual syringe. The BSA solution discharged from the first discharge port P4 through a recovery action was guided to the UV detector.

[0054] Figure 6 This is a chromatogram obtained by detecting the BSA solution recovered after aggregation in Example 1 and the BSA solution recovered without aggregation in Comparative Example 1 using a UV detector. Figure 6In the chromatogram, peak D of BSA detected in Example 1 is represented by a solid line, and peak E of BSA detected in Comparative Example 1 is represented by a dashed line. Here, the area value of peak D is 2,761,087 [μV·sec]. The area value of peak E is 2,808,307 [μV·sec]. Compared with the case where no aggregation operation was performed, the recovery rate of BSA with aggregation operation was approximately equal to 98.5%, and a high recovery rate of the target component was confirmed when the aggregation operation in the concentration apparatus 10 of this embodiment was used.

[0055] (4) Example 2

[0056] In Example 2, the same concentration apparatus 10 as in Example 1 was used to aggregate two solutions containing the same amount of the target component but with different concentrations, and the signal intensity of the chromatographic peaks was compared.

[0057] In Example 2, a BSA solution with a concentration of 1.0 mg / mL and a total volume of 100 μL (hereinafter referred to as the first BSA solution) and a BSA solution with a concentration of 0.1 mg / mL and a total volume of 1 mL (hereinafter referred to as the second BSA solution) were used. Both the first BSA solution and the second BSA solution contained 0.1 mg of BSA.

[0058] A first BSA solution is injected into the flow path FP through the third supply port P3, where it undergoes aggregation and recovery. The aggregation time for the first BSA solution is set to 1 minute. A second BSA solution is injected into the flow path FP through the third supply port P3, where it undergoes aggregation and recovery. The aggregation time for the second BSA solution is set to 2.5 minutes. Other conditions for the aggregation and recovery operations are the same as in Example 1.

[0059] Figure 7 This is a chromatogram representing the colorimetric sample obtained in Example 2. Figure 7 In the chromatogram, peak F, corresponding to BSA using the first BSA solution, is represented by a solid line, and peak G, corresponding to BSA using the second BSA solution, is represented by a dashed line. Furthermore, the dissolution times of peaks F and G shift due to differences in the aggregation time. Here, in Figure 7 In the chromatogram, the height (signal intensity) of peak F was 104146 [μV], and the height (signal intensity) of peak G was 98920 [μV]. In this case, the height of peak G was 95.0% relative to the height of peak F. Based on the results, it was confirmed that when the amount of the target component contained in the solution concentrated by the concentration device 10 is the same, peaks of approximately the same height can be detected even if the concentrations of the solutions are different.

[0060] (5) Effects of the implementation method

[0061] According to the concentration apparatus 10 of this embodiment, the target component c in the eluent accumulates on the separation membrane ME at the point where the eluent flowing in the first direction a collides with the eluent flowing in the second direction b. Furthermore, during the accumulation of the target component c, a portion of the eluent supplied from the first supply port P1 and the second supply port P2, flowing along the separation membrane ME, permeates through the separation membrane ME via tangential flow filtration. Furthermore, components in the eluent supplied from the third supply port P3 that have a size smaller than the pore size of the separation membrane ME permeate through the separation membrane ME. In this case, the concentration of the target component c can be increased without repeatedly circulating the eluent. Therefore, the target component c in the liquid can be concentrated in a short time, and a small amount of eluent containing the concentrated target component c can be recovered. For example, during the concentration operation, the target component c can be concentrated for about 1 to 2 minutes, and during the recovery operation, the eluent containing the target component c can be recovered in a volume of 2 mL or less.

[0062] Furthermore, in this embodiment, since the eluent collision position is set directly below the third supply port P3, the target component c supplied from the third supply port P3 will not disperse and aggregate in the first direction a or the second direction b. Therefore, the aggregation efficiency of the target component c is improved.

[0063] Furthermore, in this embodiment, during the recovery operation, the eluent remaining on the separation membrane ME within the flow path FP is discharged from the first discharge port P4 along with the accumulated target component c. At this time, a portion of the eluent remaining on the separation membrane ME permeates through the separation membrane ME via tangential flow filtration. Therefore, the amount of target component accumulated on the separation membrane ME can be increased, and the amount of eluent discharged from the first discharge port P4 can be reduced. In addition, since the first discharge port P4 is located at the other end of the housing 20, the amount of eluent permeating through the separation membrane ME via tangential flow filtration increases. Therefore, the concentration efficiency of the target component c is improved.

[0064] Furthermore, in this embodiment, during the recovery operation, the eluent remaining in the flow path FP and the target component c are discharged from the flow path FP through the eluent flowing in the first direction a. Therefore, the eluent containing the concentrated target component c can be recovered through simple control.

[0065] Furthermore, according to the analysis system 100 of this embodiment, a specific component in the eluent analyzed by the liquid chromatograph 1 is concentrated by the concentration device 10. Therefore, the target component in the sample analyzed by the liquid chromatograph 1 can be concentrated in a short time, and a small amount of eluent containing the concentrated target component can be easily recovered.

[0066] Additionally, an eluent containing the target component from the sample is supplied to the third supply port P3. This allows the desired component in the sample analyzed by liquid chromatograph 1 to be concentrated as target component c. Therefore, it is possible to concentrate one or more desired components from the components of a sample analyzed by liquid chromatograph 1.

[0067] (6) Other implementation methods

[0068] The concentration device 10 is included in the analysis system 100, but the concentration device 10 can also be used as a standalone unit. In this case, the supply of liquid to the first supply port P1, the second supply port P2, the first discharge port P4, and the second discharge port P5 of the concentration device 10 can be controlled, for example, using a pump, syringe, switching valve, or mass flow meter. Furthermore, the concentration device 10 of the above embodiment can be used for the concentration or desalting of proteins, peptides, and nucleic acids, as well as the purification of viruses.

[0069] The liquid supplied to the first supply port P1 and the second supply port P2 is the eluent used in the liquid chromatograph 1, but other liquids such as separately prepared ultrapure water can also be used. In this case, the eluent supplied from the third supply port P3 and the sample can be desalted. In addition, by connecting a mass analyzer or the like to the first discharge port P4, structural analysis of the target component c can be performed.

[0070] (7) Correspondence between the components of the technical solution and the parts of the implementation method

[0071] Hereinafter, examples of the correspondence between the components of the technical solution and the components of the implementation method will be described. In the implementation method, the flow regulating valve V1, pump 8a and piping p1 are examples of the first supply unit, pump 8b and piping p2 are examples of the second supply unit, switching valve V3 and piping p3 are examples of the third supply unit, and switching valve V4 and piping p4 are examples of the first discharge unit.

[0072] (8) Morphology

[0073] Those skilled in the art will understand that the various exemplary embodiments are specific examples of the following forms.

[0074] (Item 1) A concentration device of one form includes: a frame; a separation membrane that divides the internal space of the frame in such a way as to form a flow path within the frame;

[0075] The first supply section supplies the first liquid from the first position of the frame to the flow path in such a way that the first liquid flows along the separation membrane in the first direction;

[0076] The second supply section supplies the second liquid from a second position of the frame to the flow path in a manner in which the second liquid flows along the separation membrane in a second direction opposite to the first direction; and

[0077] The third supply section supplies a third liquid containing the target component with a size that does not permeate the separation membrane from the third position of the frame into the flow path.

[0078] The third position is located between the first and second positions in the first direction.

[0079] According to one type of concentration apparatus, a first liquid flowing in a first direction collides with a second liquid flowing in a second direction between a first position and a second position in the first direction along a separation membrane along a flow path. At this time, a portion of the first liquid and a portion of the second liquid permeate through the separation membrane via tangential flow filtration. In this state, a target component in a third liquid supplied to the flow path from a third position between the first and second positions accumulates at the location where the first and second liquids collide. During the accumulation of the target component, a portion of the first liquid and a portion of the second liquid flowing along the separation membrane permeate through the separation membrane via tangential flow filtration. Additionally, components in the third liquid having a size smaller than the pore size of the separation membrane permeate through the separation membrane.

[0080] In this way, the target component accumulates on the separation membrane, while components of the first, second, and third liquids, which have pore sizes smaller than the membrane, permeate through it. Therefore, the concentration of the target component can be increased without repeatedly circulating the third liquid. As a result, the target component in the liquid can be concentrated in a short time, and a small amount of liquid containing the concentrated target component can be recovered.

[0081] (Item 2) In the concentration apparatus described in Item 1, the third position may correspond to the position where the flow of the first liquid in the first direction collides with the flow of the second liquid in the second direction.

[0082] According to the concentration apparatus described in item 2, the target component is supplied to the location where the first liquid and the second liquid collide. Thus, the target component aggregates without dispersing in either the first or second direction of the first or second liquid. Therefore, the aggregation efficiency of the target component is improved.

[0083] (Item 3) The concentration apparatus described in Item 1 or Item 2 may further include a first discharge section, which discharges the first liquid, the second liquid and the third liquid from a fourth position in the flow path after the second supply section stops supplying the second liquid and after the third supply section stops supplying the third liquid.

[0084] According to the concentration apparatus described in item 3, the first liquid, the second liquid, and the third liquid remaining on the separation membrane are discharged together with the aggregated target component through the first discharge section. At this time, a portion of the first liquid, the second liquid, and the third liquid remaining on the separation membrane permeates through the separation membrane via tangential flow filtration. Therefore, the amount of target component aggregated on the separation membrane can be increased, and the amount of the first liquid, the second liquid, and the third liquid discharged from the first discharge section can be reduced.

[0085] (Item 4) In the concentration apparatus described in Item 3

[0086] The fourth position can be located further downstream than the second position in the first direction.

[0087] According to the concentration apparatus described in item 4, the first liquid, the second liquid, and the third liquid are discharged from a fourth position located further downstream than the second position. In this case, the first liquid, the second liquid, and the third liquid flow along the separation membrane in the first direction and are discharged, thus increasing the amount of the first liquid, the second liquid, and the third liquid that permeates through the separation membrane via tangential flow filtration. Therefore, the concentration efficiency of the target component is improved.

[0088] (Item 5) The concentration apparatus described in any one of items 1 to 4 may also include a control unit that controls the first supply unit, the second supply unit, the third supply unit, and the first discharge unit.

[0089] The control unit controls the first supply unit, the second supply unit, and the third supply unit to ensure that, during the first operation, the first liquid flows in the flow path in the first direction, the second liquid flows in the second direction, and the third liquid is supplied to the flow path.

[0090] The first supply section, the second supply section, the third supply section, and the first discharge section are controlled so that during the second operation after the first operation, when the second liquid and the third liquid are not supplied to the flow path, the first liquid is supplied to the flow path in a manner that flows in the first direction, and the first liquid, together with the second liquid and the third liquid remaining on the separation membrane, is discharged from the flow path.

[0091] According to the concentration apparatus described in item 5, during the first operation, the target component accumulates at the point where the first liquid and the second liquid collide. Furthermore, during the second operation, the first liquid, the second liquid, the third liquid, and the target component remaining in the flow path are discharged from the flow path. In this case, the liquid containing the concentrated target component can be recovered through simple control.

[0092] (Item 6) In the concentration apparatus described in Item 5,

[0093] In the first operation, the first supply unit supplies the first liquid to the flow path at a first flow rate, and the second supply unit supplies the second liquid to the flow path at a second flow rate greater than the first flow rate.

[0094] The third position is set closer to the first position in the first direction than the second position.

[0095] According to the concentration apparatus described in item 6, during the first operation, a first liquid flows in a first direction at a first flow rate, and a second liquid flows in a second direction at a second flow rate greater than the first flow rate. Consequently, the first liquid and the second liquid collide near a first position in the first direction. As a result, the target component accumulates at a position closer to the first position than the second position. Therefore, during the second operation, the first liquid flows from the first position in the first direction and is discharged from the fourth position, thereby increasing the distance the first, second, and third liquids remaining on the separation membrane travel along the separation membrane. This improves the efficiency of tangential flow filtration. Consequently, the concentration efficiency of the target component is improved.

[0096] (Phase 7) An analysis system, which may include:

[0097] Liquid chromatograph; and

[0098] According to any one of the concentration apparatuses in claims 1 to 6, a specific component in the eluent discharged after analysis by liquid chromatography is concentrated as a target component in a third liquid.

[0099] According to the analytical system described in item 7, a specific component in the eluent analyzed by liquid chromatography is concentrated by a concentration device. Thus, it is possible to concentrate the target component in a liquid within a short time and to recover a small amount of liquid containing the concentrated target component.

[0100] (Item 8) In the analysis system described in Item 7,

[0101] A liquid chromatograph may include:

[0102] Separation column;

[0103] The mobile phase supply section supplies the mobile phase to the separation column;

[0104] The sample inlet section introduces the sample into the mobile phase that should be supplied to the separation column by the mobile phase supply section; and

[0105] The detector measures the composition of the sample in the mobile phase derived from the separation column.

[0106] The third supply section supplies the mobile phase containing the sample discharged from the detector as a third liquid to the flow path.

[0107] According to the analytical system described in item 8, the mobile phase containing the sample can be used as a third liquid. Therefore, the components of the sample analyzed by liquid chromatography can be concentrated as target components.

[0108] (Item 9) In the analysis system described in Item 7 or Item 8,

[0109] The third supply section supplies one or more components from the sample discharged from the detector as target components, along with the third liquid, into the flow path.

[0110] According to the analytical system described in item 9, it is possible to concentrate one or more components of a sample analyzed by liquid chromatography.

[0111] (Item 10) A concentration method, comprising:

[0112] The step of supplying a first liquid from a first position in the frame to the flow path in such a way that the first liquid flows along the separation membrane in a first direction within the frame;

[0113] The step of supplying the second liquid from a second position in the frame to the flow path in a manner in which the second liquid flows along the separation membrane within the frame in a second direction opposite to the first direction; and

[0114] The step of supplying a third liquid containing a target component having a size that does not permeate the separation membrane from a third position in the housing into the flow path.

[0115] The third position is located between the first and second positions in the first direction.

[0116] In the concentration method described in item 10, the target component aggregates on a separation membrane, and components of the first, second, and third liquids, having a pore size smaller than that of the separation membrane, permeate through the membrane. Therefore, the concentration of the target component can be increased without repeatedly circulating the third liquid. As a result, the target component in the liquid can be concentrated in a short time, and a small amount of liquid containing the concentrated target component can be recovered.

Claims

1. A concentration apparatus, characterized in that, include: Frame; A separation membrane is used to divide the internal space of the frame in a manner that forms flow paths within the frame. The first supply section supplies the first liquid from a first position of the frame to the flow path in such a way that the first liquid flows along the separation membrane in a first direction; The second supply section supplies the second liquid from a second position of the frame to the flow path in such a way that the second liquid flows along the separation membrane in a second direction opposite to the first direction; as well as The third supply section supplies a third liquid containing a target component having a size that does not permeate the separation membrane from a third position within the housing into the flow path. The third position is located between the first position and the second position in the first direction.

2. The concentration apparatus according to claim 1, wherein the third position corresponds to the position where the flow of the first liquid in the first direction collides with the flow of the second liquid in the second direction.

3. The concentration apparatus according to claim 1 or 2, wherein, It also includes a first discharge section, which discharges the first liquid, the second liquid, and the third liquid from a fourth position in the flow path after the second supply section stops supplying the second liquid and after the third supply section stops supplying the third liquid.

4. The concentration apparatus according to claim 3, wherein the fourth position is located further downstream in the first direction than the second position.

5. The concentration apparatus according to claim 3, wherein, It also includes a control unit that controls the first supply unit, the second supply unit, the third supply unit, and the first discharge unit. The control unit controls the first supply unit, the second supply unit, and the third supply unit so that, during the first operation, the first liquid flows in the first direction in the flow path, the second liquid flows in the second direction, and the third liquid is supplied to the flow path. The first supply section, the second supply section, the third supply section, and the first discharge section are controlled so that, during the second operation after the first operation, when the second liquid and the third liquid are not supplied to the flow path, the first liquid is supplied to the flow path in a manner that flows in the first direction, and the first liquid, together with the second liquid and the third liquid remaining on the separation membrane, is discharged from the flow path.

6. The concentration apparatus according to claim 5, wherein during the first operation, the first supply unit supplies the first liquid to the flow path at a first flow rate, and the second supply unit supplies the second liquid to the flow path at a second flow rate greater than the first flow rate. The third position is positioned closer to the first position in the first direction than the second position.

7. An analysis system, characterized in that, include: Liquid chromatograph; as well as The concentration apparatus according to any one of claims 1 to 6 concentrates a specific component in the eluent discharged after analysis by the liquid chromatograph as the target component in the third liquid.

8. The analytical system according to claim 7, wherein the liquid chromatograph comprises: Separation column; The mobile phase supply unit supplies the mobile phase to the separation column; The sample inlet section introduces the sample into the mobile phase that should be supplied to the separation column by the mobile phase supply section; as well as The detector detects the composition of the sample in the mobile phase derived from the separation column. The third supply unit supplies the mobile phase containing the sample discharged from the detector as the third liquid to the flow path.

9. The analysis system of claim 8, wherein the third supply unit supplies the third liquid containing the target component to the flow path, using one or more components of the sample discharged from the detector as the target component.

10. A concentration method, characterized in that, include: The step of supplying the first liquid from a first position in the frame to the flow path in such a way that the first liquid flows along the separation membrane in a first direction within the frame; The step of supplying the second liquid from a second position in the frame to the flow path in such a way that the second liquid flows along the separation membrane within the frame in a second direction opposite to the first direction; as well as The step of supplying a third liquid containing a target component having a size that does not permeate the separation membrane from a third position of the housing into the flow path. The third position is located between the first position and the second position in the first direction.

Images