Filtering system and method thereof

Abstract

The invention relates to the technical field of filtration of biological fluids and products, in particular to a filtration system and a method thereof. A first filter unit having a first end and a second end and in fluid communication with the processing vessel; a second filter unit having a first end and a second end and in fluid communication with the processing vessel; the driver is used for generating a magnetic field; an alternating flow pump connected to a first end of the first filter unit and a second end of the second filter unit, the alternating flow pump having a mover fully sealed within a pump chamber, the mover being capable of generating a magnetic field coupled to the driver magnetic field, and the driver drives the rotor to reciprocate in the pump chamber by controlling the change of the magnetic field so as to pump fluid from the processing container through the first filter unit and the second filter unit.

Description

Technical Field

[0001] The present invention relates to the technical field of filtering biological fluids and products, and in particular to a filtering system and a method thereof. Background Art

[0002] 1. Filtration and cell culture Filtration is commonly used to separate, clarify, modify and / or concentrate fluid solutions, mixtures or suspensions. In the biopharmaceutical and medical industries, filtration is essential to the successful production, processing and analysis of drugs, diagnostics, chemicals and many other products. As examples, filtration can be used to sterilize liquids or gases, to clarify complex suspensions into a filtered "clear" fraction and an unfiltered fraction; similarly, components in a suspension can be concentrated by removing or "filtering out" the suspending medium. Further, many other specialized filter applications have been developed through appropriate selection of filter materials, filter pore size or other filter variables; these applications can include the selective separation of components from different sources, including biological cultures, blood and other fluids that can be solutions, mixtures or suspensions. With the continuous advancement of cell and recombinant DNA technology, many new products have been developed, many of which have such complex compositions and mechanisms of action that they can only be produced using cell culture technology, through the complex synthetic mechanisms of living cells, for example, proteins, receptors, vaccines and antibodies for treatment, research and diagnosis, as well as cells themselves. By maintaining a high cell concentration and high viability culture at a high yield over a long period of time, a preliminary target product can be obtained. The use of filtration is an effective means that can be used to increase the yield of cell culture.

[0003] Biologics manufacturing processes have advanced through substantial process intensification. Eukaryotic and prokaryotic cell cultures currently used to produce recombinant proteins, antibodies, virus-like particles (VLPs), gene therapy particles, and vaccines now include cell expansion levels of 10E7 cells / mL or higher. This is achieved through the use of cell retention devices that are used to remove metabolic waste products and provide nutrient renewal support. The most common methods of cell retention include the use of hollow fiber membrane filtration with alternating tangential flow (ATF) to perfuse bioreactor cultures and the use of "dead-end flow filtration" using flat membranes with multilayer structures to perfuse bioreactor cultures.

[0004] (II) Filtration method Filter chemistries, configurations, and morphologies have been developed to facilitate the separation of materials based on their chemical and physical properties; despite the widespread development of filter technology, they are generally limited by their tendency to clog: for example, when filters are used to filter suspensions of cultured mammalian cells, the filters tend to clog with dead cells, cell debris, aggregates, fibrous biomolecules, or other components present in the complex culture "soup". In this regard, the filtration method can have a profound impact on filtration efficiency and membrane life. In one filtration method, commonly referred to as "dead-end" filtration, the entire fluid passes through the membrane perpendicular to the membrane surface. Debris accumulates rapidly on the surface, causing the membrane to become rapidly clogged. Typically, applications that utilize dead-end filtration involve small samples. Another filtration method, commonly referred to as tangential flow filtration (TFF), improves on dead-end filtration. In TFF, the fluid to be filtered is recirculated by a pump, typically from a reservoir through the filter and back to the reservoir. The flow through the filter is parallel to the surface of the filter. Debris accumulation is effectively removed by the flushing action of the circulating fluid. However, one of its limitations is the tendency to form gel-like deposits on the filter surface, which still limits the effectiveness of the filter and eventually causes it to clog. Another method, called alternating tangential flow filtration (ATF), provides another filtration model: it is similar to TFF in that it produces a flow pattern parallel to the filter membrane surface; however, it differs from TFF in that the flow direction across the filter surface is constantly alternating.

[0005] 3. Limitations of the existing solutions U.S. Patent No. 6,544,424 discloses a perfusion system in which during the suction stroke, a diaphragm pump draws culture fluid from a bioreactor through a hollow fiber filter, and during the discharge stroke, the liquid is pushed back to the bioreactor through the filter. During the suction stroke, permeation occurs, and during the discharge stroke, the filter is flushed with a backflow to reduce blockage caused by cells. Other developments of this technology are described in WO2012 / 026978, which relates to the incorporation of a bioreactor and a perfusion unit into a single device, and in US 2011 / 0111486, in which the flow to and from the perfusion unit is also used to provide additional stirring in the bioreactor.

[0006] However, the filtration efficiency of these systems is not optimal because the diaphragm pump uses a vacuum on the underside of the diaphragm during the "pumping" cycle to draw liquid from the process vessel down through the filter. Although this system applies positive pressure up to 3.5 bar, the maximum "vacuum" that can be applied is about -1 bar. This vacuum can be further affected by losses in the tubing and components between the vacuum source and the diaphragm pump. In addition, if the viscosity of the fluid changes, a greater negative pressure behind the diaphragm may be required to obtain the full displacement of the pump. All of these factors can reduce the efficiency of conventional pumping systems.

[0007] And therefore, the hollow fiber filter is not used evenly along its entire length during a cycle, making it susceptible to clogging and fouling of the membrane.

[0008] Another problem with using a diaphragm pump to control flow through a filter is that it requires a compressed gas source and vacuum equipment, including associated hoses and power units, in order to actuate the pump. Additionally, because the activation gas (e.g., air) is compressible, there is an undesirable activation delay until sufficient pressure builds up in the gas phase chamber to push the diaphragm upward to move the fluid out of the liquid phase chamber and through the filter. This activation delay can adversely affect overall process efficiency.

[0009] In addition, diaphragm pumps use positive and negative air flows to provide liquid delivery power. However, conventional systems cannot accurately control the change between positive and negative air flows or the amount of positive and negative air flows. Moreover, such systems usually have complex structures that are difficult to manufacture and maintain and include airflow restrictions.

[0010] More importantly, large-scale production using diaphragm pump devices requires the ability to withstand large forces and maintain diaphragm integrity. However, diaphragms are inherently limited by their material lifespan, and in long-term culture, there is often a failure in diaphragm integrity that causes the system to fail.

[0011] Therefore, a filtration system is provided that fundamentally eliminates the integrity defects of diaphragm pumps, while eliminating the limitations of gas compressibility and vacuum. When the system is in operation, the pressure can be precisely controlled during the cycle, ultimately achieving improved robustness and overall efficiency of the perfusion culture system, and reducing system complexity and cost.

[0012] (IV) Cell culture scale-up and scale-down processes The use of animal cell culture is increasingly used for the production of different cell-derived biologicals, which can be natural or manufactured, including proteins, nucleic acids, metabolites and many other substances. Therefore, the production methods may also be different. The production methods can range from "simple" batch to continuous processes. In the ordinary batch culture production process, cells are first inoculated into fresh culture medium, after which the cells enter an exponential growth phase. As they consume nutrients in the culture medium, waste accumulates; concomitantly, the cells switch from rapid growth to a stationary growth phase, and then enter a cell decay phase. Although several methods for optimizing batch culture products have been developed, in each case, these processes still undergo rapid growth and decay cycles. Another culture process involves continuous use of a method commonly known as perfusion to maintain the culture. In the perfusion culture method, while retaining the cells, the waste produced by the cells is continuously removed from the culture, and the removed waste culture medium is supplemented with fresh culture medium. Using this method, it is possible to achieve that the cell concentration and productivity are kept in a balanced state. Typically, one to two culture volumes are exchanged per day, and the cell concentrations achieved in the perfusion method are typically 2 to more than 10 times the cell concentrations achieved at the peak of batch cultures. It has long been believed that perfusion culture provides relatively good economics for cell culture. Perfusion culture operations can achieve high cell densities, and more importantly, cells can be maintained in a high output state for several weeks. This achieves much greater output and reduces the necessary bioreactor volume. It is also a useful technique for culturing slow-growing cells.

[0013] However, despite the benefits of perfusion methods, their acceptance has been slow, and one reason for this slow acceptance may be that the vast majority of products are derived on a small scale from batch culture systems, such as "T" bottles. If more material is needed, it is usually produced by increasing the number and size of "T" bottles or transferring the culture to roller bottles or spinner bottles, both of which are also batch cultures in nature. When the culture is scaled up to a bioreactor, the process has been greatly affected by the previous handling of the culture. Attempts to address this problem using hollow fiber bioreactors or other fixed bed bioreactors, in which cells gradually attach or become entrapped to a fixed surface, have been only partially effective; the inherent heterogeneity and inconvenience to the cells of such reactors limit their usefulness as research tools.

[0014] Therefore, a perfusion system that can be easily scaled down is provided, wherein the filtration system should have a smaller liquid phase chamber compared to other systems while maintaining the same level of displacement so as to make the perfusion system easier to use on a small scale and at the research and development level.

[0015] The information disclosed in this background technology section is only intended to deepen the understanding of the overall background technology of the present invention, and should not be regarded as acknowledging or suggesting in any form that the information constitutes the prior art already known to those skilled in the art. Summary of the invention

[0016] The object of the present invention is to provide a filtering system and a method thereof to solve the technical problems existing in the prior art.

[0017] In order to achieve the above object, the present invention adopts the following technical solutions: The present invention provides a filtering system, comprising: Handling containers; a first filter unit having a first end and a second end and being in fluid communication with the processing vessel; a second filter unit having a first end and a second end and being in fluid communication with the processing vessel; A driver for generating a magnetic field; An alternating flow pump is connected to the first ends of the first filter unit and the second filter unit, and has a mover that is completely sealed in a pump chamber. The mover can generate a magnetic field that couples with the magnetic field of the driver. The driver drives the mover to reciprocate in the pump chamber by controlling changes in the magnetic field, so as to pump the fluid from the processing container through the first filter unit and the second filter unit.

[0018] Preferably, the alternating flow pump generates an alternating tangential flow of the fluid inside the first filter unit and the second filter unit.

[0019] Preferably, the mover has a sealing device to divide the pump chamber of the alternating flow pump into a first chamber and a second chamber; when the mover moves toward the first end direction away from the first filter unit, that is, the α stroke, the alternating flow pump sucks the fluid in the first filter unit and the processing container into the first chamber of the alternating flow pump; at the same time, the fluid in the second chamber of the alternating flow pump is discharged into the second filter unit and the processing container; when the mover moves toward the first end direction away from the second filter unit, that is, the β stroke, the alternating flow pump sucks the fluid in the second filter unit and the processing container into the second chamber of the alternating flow pump; at the same time, the fluid in the first chamber of the alternating flow pump is discharged into the first filter unit and the processing container.

[0020] Preferably, the coupling between the magnetic field of the mover of the alternating flow pump and the magnetic field of the driver includes concentric coupling, end surface coupling, or a composite coupling.

[0021] Preferably, the change of the driver magnetic field includes a change in magnetic field flux, a magnetic field displacement, and a magnetic pole conversion.

[0022] Preferably, the sealing device is a liquid sealing device arranged on the mover.

[0023] Preferably, the device further comprises: a controller connected to the driver and configured to control the movement of the driver so as to controllably move the mover.

[0024] Preferably, it further comprises: a first end pressure sensor, the first end pressure sensor being arranged to measure the pressure between the first end of the first filter unit and the second filter unit and the alternating flow pump chamber; and / or, A second end pressure sensor is arranged to measure pressure between the second ends of the first and second filter units and the process vessel.

[0025] The controller is arranged to control the driver to control the alternating flow pump based on pressure data received from at least one of the first end and second end pressure sensors of the first filter.

[0026] Preferably, it further comprises: a linear encoder connected to the driver for determining the position of the movable driver.

[0027] Preferably, the first filter unit and the second filter unit are each connected to a pipeline branch point via a first permeate pipeline and a second permeate pipeline, and the pipeline branch point is connected to a permeate pump.

[0028] Preferably, the alternating flow pump generates alternating flow in the first permeate pipe and the second permeate pipe, and part of the permeate is returned from the first permeate pipe and the second permeate pipe to the retentate chamber of the filter through the permeate chamber of the filter, thereby backwashing the filter membrane.

[0029] Preferably, when the mover moves toward the first end direction away from the first filter unit, that is, the α stroke, the alternating flow pump sucks the fluid in the first filter unit and the processing container into the first chamber of the alternating flow pump, and the fluid passes through the first permeate pipe and returns to the retentate chamber of the first filter, forming backwashing of the filter membrane; at the same time, the fluid in the second chamber of the alternating flow pump is discharged into the second filter unit and the processing container, and permeation is formed; when the mover moves toward the first end direction away from the second filter unit, that is, the β stroke, the alternating flow pump sucks the fluid in the second filter unit and the processing container into the second chamber of the alternating flow pump, and the fluid passes through the second permeate pipe and returns to the retentate chamber of the second filter, forming backwashing of the filter membrane; at the same time, the fluid in the first chamber of the alternating flow pump is discharged into the first filter unit and the processing container, and permeation is formed.

[0030] Preferably, the first end of the first filter unit is connected to the processing container via a first end check valve, and the second end of the first filter unit is connected to the processing container via a second end check valve; the first end of the second filter unit is connected to the processing container via the first end check valve; the second end of the second filter unit is fluidly connected to the processing container via the second end check valve; the first end check valve is arranged to allow fluid flow in the direction from the processing container to the first end and hinder flow in the opposite direction; the second end check valve is arranged to allow fluid flow in the direction from the second end to the processing container and hinder flow in the opposite direction.

[0031] Preferably, it also includes: the first end check valve and the second end check valve of the first filter unit are respectively connected to a pipeline branch point, and the pipeline branch point is further connected to the processing container via a pipeline section; a first end control valve is arranged between the first end of the first filter unit and the pipeline branch point, and a second end control valve is arranged between the second end of the first filter unit and the branch point.

[0032] Preferably, it also includes: the first end check valve and the second end check valve of the second filter unit are respectively connected to a pipeline branch point, and the pipeline branch point is further connected to the processing container via a pipeline section; a first end control valve is arranged between the first end of the second filter unit and the pipeline branch point, and a second end control valve is arranged between the second end of the second filter unit and the pipeline branch point.

[0033] Preferably, the alternating flow pump comprises: a main alternating flow pump connected to the first end of the first filter unit and the second filter unit; at least one extended alternating flow pump connected to the first end of the first filter unit and the second filter unit through a manifold; The driver comprises: A main driver, the main driver being adapted to the main alternating flow pump; At least one expansion drive adapted to the expansion alternating flow pump.

[0034] Preferably, the alternating flow pump comprises: a main alternating flow pump connected to the first end of the first filter unit and the second filter unit; at least one gain alternating flow pump connected to the first filter unit and the second end of the second filter unit; The driver comprises: A main driver, the main driver being adapted to the main alternating flow pump; At least one gain driver, the gain driver is adapted to the gain alternating current pump.

[0035] The present invention provides a filtering method, which comprises the following steps: S1. adding cell culture medium and cells to the bioreactor, wherein the reactor type includes but is not limited to a stirred reactor, a loop reactor, a wave reactor, etc.; S2. culturing cells in the bioreactor; S3. During the culture period, operating the alternating flow pump, comprising: S31, applying positive pressure to the first end of the first filter unit, discharging the fluid from the pump chamber to the bioreactor chamber via the second end of the first filter unit and the pipe section, during which a portion of the fluid is guided to enter the permeate chamber of the first filter unit via the semi-permeable wall portion of the first filter unit; and simultaneously, applying negative pressure to the first end of the second filter unit, sucking the fluid from the bioreactor chamber to the pump chamber via the pipe section and the second end of the second filter unit, during which a portion of the fluid is guided to enter the permeate chamber via the semi-permeable wall portion of the second filter unit; S32, and reverse the direction of the force applied by the alternating flow pump, so that the second end of the first filter unit draws fluid from the bioreactor chamber through the pipe segment, and at the same time, the second end of the second filter unit discharges fluid to the bioreactor chamber through the pipe segment.

[0036] S33, repeating steps S31 and S32 at least once, wherein the permeate chamber, the reactor chamber and the alternating flow pump chamber are part of the same closed filtration system; S4. Aspirating permeate from the permeate outlet port. By adopting the above technical solution, the present invention has the following beneficial effects: The present invention eliminates the risk of integrity failure of the diaphragm of the diaphragm pump and improves the efficiency of the pump system and the life of the filtration system. The system of the present invention is scalable and has a smaller retentate chamber volume (including retentate pipeline, filter retentate chamber and pump chamber) while maintaining the same displacement as other disclosed filtration systems. The latter is crucial for scaled-down models and research. It is compatible with shaking bioreactors such as wave-type and orbital oscillation-type bioreactors and various stirred reactors. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0038] Figure 1 A schematic diagram of a filtration system provided in Embodiment 1 of the present invention; Figure 2 A first schematic diagram of the relationship between the movement direction of the alternating flow pump and the fluid discharge and fluid intake of the filter unit provided in the first embodiment of the present invention; Figure 3 A second schematic diagram of the relationship between the movement direction of the alternating flow pump and the fluid discharge and fluid intake of the filter unit provided in the first embodiment of the present invention; Figure 4 A schematic diagram of the location of the bubble trap in the filtration system provided in the first embodiment of the present invention; Figure 5 A schematic diagram of the periodic change of the alternating flow pump provided in the first embodiment of the present invention; Figure 6 A schematic diagram of a liquid sealing device for a mover provided in Embodiment 1 of the present invention; Figure 7 A three-dimensional diagram of a sealed permanent magnet device of a mover provided in Embodiment 1 of the present invention; Figure 8 A cross-sectional view of a sealed permanent magnet device of a mover provided in Embodiment 1 of the present invention; Fig. 9 A schematic diagram of the assembled driver and alternating flow pump provided in the first embodiment of the present invention; Fig.10 This is a schematic diagram of the internal structure of the driver and alternating flow pump after assembly provided in the first embodiment of the present invention; Fig.11 A schematic diagram of a driver and a mover provided in Embodiment 1 of the present invention in which the magnetic coupling mode is concentric coupling; Fig.12 A schematic diagram of the first embodiment of the present invention showing that the magnetic coupling mode between the driver and the mover is end-face coupling; Fig.13 A schematic diagram of a perfusion system with a controller and a sensor provided in Embodiment 2 of the present invention; Fig.14 A schematic diagram of a filtration system provided in Embodiment 3 of the present invention; Fig.15 A schematic diagram of a filtration system provided in Embodiment 4 of the present invention; Fig.16 A schematic diagram of a filtration system provided in Embodiment 5 of the present invention; Fig.17 This is a schematic diagram of a filtration system provided in Example 6 of the present invention.

[0039] Icons: 1-first filter unit; 2-second filter unit; 3-first end; 5-first pump chamber; 6-reciprocating motion direction of driver magnetic field; 7-driver magnetic field; 8-mover magnetic field; 9-mover; 10-second pump chamber; 11-alternating tangential flow; 12-second end; 13-valve; 14-sterile sealed pipeline; 17-permeate pump; 18-mixing device driver; 19-fluid pump; 22-bioreactor; 23-bioreactor chamber; 25-mixing device; 26-bubble trap device; 27-liquid sealing device; 28-sealed permanent magnet; 29-permanent magnet diaphragm; 30-driver housing; 31-alternating flow pump; 32-driver guide rail; 33-linear drive mechanism; 34-driver permanent magnet; 35-driver permanent magnet diaphragm; 36-pressure sensor; 37-linear encoder; 38-liquid level sensor; 39-controller. DETAILED DESCRIPTION

[0040] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0041] The specific implementation of the present invention is described in detail below in conjunction with the accompanying drawings. It should be understood that the specific implementation described here is only used to illustrate and explain the present invention, and is not used to limit the present invention. Example

[0042] Combination Figures 1 to 12 As shown, the first embodiment provides a duplex mode fluid filtration system, which uses a mover driven by magnetic force to perform two-phase pumping in a sterile sealed container to create an alternating tangential flow 11 (mode A system) in the filter unit.

[0043] The system includes: Handling containers; a first filter unit 1 having a first end 3 and a second end 12 and being in fluid communication with the processing vessel; a second filter unit 2 having a first end 3 and a second end 12 and being in fluid communication with the processing vessel; a driver comprising a magnetic field, and An alternating flow pump 31 is connected to the first end of the first filter unit 1 and the second filter unit 2, and has a mover 9 completely sealed in a pump chamber, and the mover 9 has a device that can generate a magnetic field, which can be coupled with the magnetic field of the driver and driven by the change of the driver magnetic field 7. The alternating flow pump 31 can be operated to pump the fluid from the processing container through the first filter unit 1 and the second filter unit 2.

[0044] In this embodiment, the processing container may be a bioreactor.

[0045] For example: the bioreactor includes a funnel-shaped container for cell culture; the funnel-shaped container is a vessel structure with a funnel-shaped bottom and a top cover; the bottom tip of the funnel-shaped container is set as a spherical arch structure or a flat bottom structure; a supporting device is set under the funnel-shaped container, and the supporting device is connected to an eccentric driving device.

[0046] Preferably, the bottom cone angle a of the funnel-shaped container is maintained between 60-150 degrees.

[0047] It is understandable that the shape of the container is not limited thereto, and can be adaptively deformed according to actual needs. For example, the cross-section of the container can be a polygon, or other similar shapes, all of which fall within the protection scope of the present application.

[0048] Preferably, a supporting device is provided below the funnel-shaped container, the upper end of the supporting device is connected to the outer side wall of the funnel-shaped container, and the lower part of the supporting device is connected to the eccentric driving device.

[0049] Preferably, the funnel-shaped container is made of a rigid material, which is suitable for containers of small volume.

[0050] Preferably, the funnel-shaped container is made of a flexible material, and a shape-retaining support body is provided around the funnel-shaped container, which is suitable for large-volume containers.

[0051] The funnel-shaped container may be equipped with a flow-turbulating device, which can be used to increase the mixing effect and adjust the flow field environment, such as shear force, turbulent kinetic energy dissipation rate, etc. The number and form of the flow-turbulating device can be designed according to specific process requirements.

[0052] Preferably, when the container is made of a rigid material, the spoiler can be directly welded to the inner wall of the container.

[0053] Preferably, when the container is made of a flexible material, the spoiler can be mounted on a shape-maintaining support around the container (as shown in the figure).

[0054] Preferably, the eccentric drive device comprises: a shaking table, a six-axis robot, or a combination structure of a drive motor and a drive motor and an eccentric amplitude adjustment device.

[0055] Preferably, the circulating flow bioreactor further comprises: a control system, wherein the control system is communicatively connected to the sensor detection element arranged on the funnel-shaped container.

[0056] Specifically, the container can be connected to a control system, and the values ​​of various process parameters in the container can be collected through sensors configured on the container. Through the control program, in conjunction with pumps, flow meters and other devices, automatic control of physical parameters in the container can be achieved, including but not limited to temperature, rotation speed, PH, DO, gas flow, etc.

[0057] The bioreactor uses an eccentric drive device with a certain eccentric amplitude, combined with a funnel-shaped container with a unique geometric design, to provide a gentle flow field environment for cell growth, increase cell culture density, and achieve planned culture.

[0058] In this embodiment, the second end of the first filter unit 1 is connected to the bioreactor chamber via a pipeline segment (eg, a sterile sealed pipeline 14 with a valve 13 disposed therein), and the first end thereof is connected to the first chamber of the alternating flow pump 31 .

[0059] In this embodiment, similarly, the second end of the second filter unit 2 is connected to the bioreactor chamber 23 via a pipeline segment and is arranged at an end opposite to the first filter, and its first end is connected to the second chamber of the alternating flow pump 31 .

[0060] In this embodiment, compared to other systems that use one pipe to connect the reactors, the present system makes the diameter of the pipe required in the bioreactor smaller and increases the flexibility of the pipe around the bioreactor. The latter is particularly important for bioreactors that move during the culture process (for example, wave reactors, orbital oscillation reactors, and torrent reactors, etc.), where the pipe moving with the reactor requires better flexibility to reduce its length to minimize the retentate capacity and pressure drop of the pipe section. At the same time, the liquid in the pipe between the bioreactor and the first and second filters is processed separately in a single pass, and the ratio between the processed volume and the stagnation volume in the pipe between the bioreactor and the filter is reduced, which is very important for scale-up, especially scale-down models.

[0061] In this embodiment, in some embodiments, the processing container is a bioreactor. Figure 1 As shown, the bioreactor may suitably include a container 22 having an internal volume and a stirring device (mixing device driver 18 and mixing device 25) positioned inside and / or outside the internal volume. The stirring device may, for example, be an impeller inside the internal volume, which is directly or magnetically driven by a motor outside the internal volume. The stirring device may also be a movable support on which the container is placed, which can transmit shaking and / or swinging motion to the container, so that the liquid phase and gas phase space produce mixing and mass transfer effects. In addition, the specific form of the bioreactor 22 is not limited, and may also include: a fluid pump 19 and the like.

[0062] In this embodiment, the filter unit can be a filter suitable for tangential flow filtration, for example, a hollow fiber cartridge or a flat plate cassette having a plurality of membranes defining a retentate side and a permeate side. Figure 1 As shown, the retentate passes through the hollow fiber chamber from the first end of the filter unit through the pipeline section, and enters the alternating flow pump 31 chamber through the other end of the hollow fiber bundle, which is the retentate chamber. The permeate outlet port can be connected to the permeate chamber on the shell side surrounding the hollow fiber bundle. The hollow fiber filter can also include two permeate outlet ports, in which case it is possible to circulate the permeate along the hollow fiber in the permeate chamber in a direction that is countercurrent (opposite) to the retentate circulation direction. The filter can be contained in an elongated (e.g., cylindrical) housing, wherein the hollow fiber bundle is positioned in the central portion of the housing, and a seal (not shown) demarcates the permeate chamber from the retentate chamber. The fluid will enter the permeate chamber through the semi-permeable wall of the hollow fiber and be discharged from the system at a certain flow rate via the permeate pump.

[0063] In this embodiment, it is also possible to use a flat panel box of a conventional tangential flow filtration type. In this case, the retentate inlet orifice corresponds to the fiber cavity end at the second end, the retentate outlet orifice corresponds to the cavity end at the first end, and the permeate orifice corresponds to the permeate outlet.

[0064] In this embodiment, the duplex mode alternating flow pump 31 may appropriately include a reciprocating component, such as a diaphragm or a mover. The reciprocating component may move back and forth relative to the pump chamber. The duplex mode alternating flow pump 31 may be, for example, Figure 5 The cylindrical shape shown in the figure completely seals the mover in the pump chamber, wherein the pump chamber is separated from the mover with a sealing device, and the mover is a reciprocating moving part. The mover also has a device for generating a magnetic field, which can be coupled with the driver magnetic field 7 outside the sterile sealed pump chamber. The magnetic field of the driver can change, such as magnetic field displacement (drive magnetic field reciprocating direction 6), magnetic field intensity change, and magnetic pole change, etc., to drive the mover coupled thereto to reciprocate in the sterile sealed pump chamber. During the stroke, the stroke volume of the duplex mode alternating flow pump 31 corresponds to the volume of the fluid (culture fluid) transferred from or into the pump chamber. The pump chamber can be directly connected to the retentate chamber of the filter, for example, at both ends. Alternatively, it may be connected via a fluid connector (not shown) having a diameter large enough not to impede liquid flow and a volume significantly smaller than the stroke volume of the alternating flow pump 31 (e.g., less than 20% of the stroke volume, such as less than 10% or less than 5% of the stroke volume), optionally via a sterile connector.

[0065] In this embodiment, Figure 5 As shown, during the movement of the mover of the alternating flow pump 31, positive pressure is applied to the fluid in one of the pump chambers, while the other pump chamber of the alternating flow pump 31 is at negative pressure. Therefore, during the operation of the alternating flow pump 31, both a suction stroke (negative pressure) and a discharge stroke (positive pressure) are included.

[0066] In this embodiment, when the mover moves in the direction away from the first end of the first filter unit 1 (α stroke), the alternating flow pump 31 generates a suction stroke (negative pressure) in the first pump chamber 5, and the fluid (for example, culture fluid) is sucked from the processing container through the first filter unit 1 through the pipeline into the first chamber of the alternating flow pump 31 in the pump chamber; at the same time, a discharge stroke (positive pressure) is generated in the second pump chamber 10, forcing the fluid (for example, culture fluid) to be discharged from the second pump chamber 10 through the second filter unit 2 through the pipeline into the processing container, and the culture fluid is pushed through the retentate side of the second filter, thereby generating a pumpable permeate.

[0067] In this embodiment, when the mover moves in the direction away from the first end of the second filter unit 2 (β stroke), the alternating flow pump 31 generates a suction stroke (negative pressure) in the second pump chamber 10, and the fluid (for example, culture fluid) is sucked from the processing container through the second filter unit 2 through the pipeline into the second chamber of the alternating flow pump 31 in the pump chamber; at the same time, a discharge stroke (positive pressure) is generated in the first pump chamber 5, forcing the fluid (for example, culture fluid) to be discharged from the first pump chamber 5 through the first filter unit 1 through the pipeline into the processing container, and the culture fluid is pushed through the retentate side of the first filter, thereby generating a pumpable permeate.

[0068] In this embodiment, during the α stroke and the β stroke of the alternating flow pump 31 , the forces applied to the mover are opposite, and the phases of the fluid flows in the first filter unit 1 and the second filter unit 2 are opposite.

[0069] In this embodiment, in some embodiments, the sealing device of the mover is a liquid sealing device 27, such as Figure 6 As shown, it can generate a liquid seal layer on the outer surface of the mover through the pipe at the front end of the mover in the positive pressure pump chamber. The liquid seal layer can reduce the friction force on the mover when the mover moves. In the long-period reciprocating motion, it can not only reduce the strength requirements of the magnetic coupling and avoid the risk of demagnetization, but also reduce the fluid heating phenomenon caused by the friction effect, thereby reducing the impact on fragile materials in the fluid such as animal cells and biological molecules.

[0070] In this embodiment, the magnetic field of the mover can be a constant magnetic field, such as a sealed permanent magnet 28 and a permanent magnet spacer 29 arranged between the sealed permanent magnets 28, which can be coupled with the magnetic field of the driver outside the pump chamber. The magnetic coupling method can be concentric coupling, end face coupling, or a composite form of coupling. The number of magnetic couplings can be appropriately increased in the movement direction (axial direction) of the mover, and the magnetic poles of the magnets in the axial direction of the mover should be arranged in adjacent opposite directions to increase the coupling strength between the mover magnetic field 8 and the driver magnetic field 7 and reduce the risk of axial demagnetization.

[0071] In this embodiment, Figure 4 As shown, a bubble trap device 26 is provided between the housing of the alternating flow pump 31 and the first filter unit 1 and the second filter unit 2 to separate a small amount of bubbles mixed in the liquid due to factors such as raw materials, processes, pressure fluctuations, empty pipes, etc. during the liquid material transportation process. The bubble trap can be directly connected to the pipe between the filter and the pump chamber, for example. Alternatively, it can be connected via a fluid connector (not shown), optionally via a sterile connector.

[0072] In this embodiment, the magnetic field of the driver can be a constant magnetic field, such as a driver permanent magnet 34 and a driver permanent magnet spacer 35 arranged in the driver housing 30. The driver further includes a linear drive mechanism 33 and a driver guide rail 32, which can drive the mover in the pump chamber to perform precisely controlled reciprocating linear motion.

[0073] In this embodiment, the construction materials used in the system can be appropriately compatible with commonly used sterilization methods (e.g., gamma irradiation and / or autoclaving). For reusable components, stainless steel (e.g., having a corrosion resistance at least equal to 316L) or engineered plastics (e.g., polysulfone, PEEK, etc.) can be used, while for single-use components, plastics (e.g., polysulfone, polypropylene, polyethylene, or ethylene copolymers) can be used.

[0074] In this embodiment, in order to avoid infection of the cell culture, the bioreactor and all components in contact with the culture medium are appropriately sterilized before cultivation. The system or parts of the system can be assembled and sterilized by autoclaving or radiation, or one or more components can be pre-sterilized and assembled in a sterilization system. To facilitate assembly, the sterilization system parts or components can be equipped with sterile connectors, for example, CPC's AseptiQuik type. Alternatively, the sterilization system parts / components can be contained in sterile packaging or assembled in a sterilization clean room.

[0075] In this embodiment, the permeate outlet port is connected to a permeate pump 17. The permeate pump 17 can be used to draw permeate from the filter and can also be used to control the transmembrane pressure in the filter unit. The permeate pump can be, for example, a peristaltic pump. The permeate pump 17 can be arranged to operate in two flow directions, for example in a forward direction for conveying permeate from the permeate outlet to a reservoir or waste container (not shown) or to further processing operations (not shown), and in a backward direction for backwashing the filter membrane with permeate.

[0076] In this embodiment, Figure 7 In certain embodiments shown, the system includes a first end pressure sensor 36 arranged to measure pressure at the first end of the filter. The pressure sensor 36 can be positioned at the first end of the filter or at any point between the first end and the pump chamber. The system may also (or alternatively) include a pressure sensor 36 arranged to measure pressure at the second end. The pressure sensor 36 may be positioned at the second end of the filter unit or at any point between the second end and the reactor chamber. The pressure sensor 36 may be a manometer or sensor that can be read manually, which is arranged to transmit pressure data to the controller 39, for example, by electrical, electromagnetic or optical devices. The second end sensor can also be used, for example, for accurate determination of the transmembrane pressure on the filter unit and the membrane.

[0077] In this embodiment, the permeate outlet port includes an osmotic pressure sensor 36. The osmotic pressure sensor 36 may be capable of measuring the pressure on the permeate side of the filter, which may be combined with data from the first end and / or second end pressure sensor 36 to calculate the transmembrane pressure on the filter unit. The osmotic pressure sensor 36 may also be used in a feedback loop for controlling the transmembrane pressure through the operation of the osmotic pump or through other osmotic pressure controllers 39 (e.g., hydrostatic pressure control). The pressure sensor 36 is also useful for control of the filter membrane backwash cycle during the stroke of the alternating flow pump 31.

[0078] In this embodiment, a pressure sensor 36 at the first end of the second filter unit 2 which is optionally connected to the controller 39 is also included, and a second end pressure sensor 36 is optionally included.

[0079] In this embodiment, Figure 8 In some of the embodiments shown, the alternating flow pump 31 is a magnetically driven pump, and the system includes a controller 39 connected to the driver, the controller 39 is used to control the movement of the driver to controllably move the mover, and the alternating flow pump 31 can apply positive pressure to one of the pump chambers separated by the mover seal device during the cycle while applying negative pressure to the other pump chamber, so that the filtration process and transmembrane pressure can be effectively measured and controlled by using a mechanical sensor (not shown) of the driver, and the mechanical sensor is not in contact with the biological fluid. This design is advantageous when designing a system with a single-use component in contact with the biological fluid. It reduces cost and complexity because the driver and mechanical sensor are reusable and do not need to be sterilized. Therefore, non-sterilizable high-performance sensors can also be used, and if sterilizable high-performance sensors are used, they do not need to be recalibrated after sterilization.

[0080] In this embodiment, Figure 8In certain embodiments shown, the system includes at least one controller 39 arranged to control the alternating flow pump 31 based on pressure data received from at least one of the inlet and outlet pressure sensors 36. The controller(s) 39 may be suitably electrically, electromagnetically (e.g., by wireless communication), optically (e.g., via optical fiber) or pneumatically connected to at least one of the first and second end pressure sensors 36. The controller(s) 39 may be a computer, a programmable logic controller 39 or any similar device capable of a) receiving input signals from one or more pressure sensors 36; b) calculating one or more output parameters from the input signals according to a predetermined method, and c) transmitting the output parameter(s) as a signal / signals to one or more control valves and / or pumps. The controller 39 may be an integral controller 39 arranged to control both the alternating flow pump 31 and the inlet and / or outlet control valves, or the system may include a main controller 39 (or valve controller 39) and a pump controller 39 (not shown). The pump controller 39 may, for example, be arranged to control the flow and pressure profile produced by the alternating flow pump 31 by one or more of the stroke frequency, stroke length and velocity of the moving parts during the α and β strokes. As mentioned above, features of the pump controller 39 may also be incorporated into the main controller 39, or vice versa, and the control valve(s) may also be controlled by the overall pump controller 39. Alternatively, the control valve(s) may also be pre-set or manually adjusted so that only one pump controller 39 is required.

[0081] In this embodiment, the controller 39 may also be connected to the osmotic pump and / or the osmotic pressure sensor 36 for controlling the transmembrane pressure.

[0082] In this embodiment, a linear encoder 37 or another stroke measurement device can be coupled to the actuator for determining the position of the actuator. A controller 39 can be coupled to the actuator and can control the actuator to move the actuator in a controllable manner. The controller 39 includes a processor that executes instructions for controlling the stroke distance and stroke rate of the actuator. The controller 39 can also include a memory associated with the processor, the memory including predetermined stroke distance and stroke rate values ​​associated with specific filter and pump sizes.

[0083] In this embodiment, a weight sensor optionally connected to the controller 39 is included, which is arranged at the lower end of the processing container to measure the weight change of the processing container to control the amount of fluid (e.g., cell culture medium) added to the bioreactor and the amount of permeate sucked from the permeate outlet port during cultivation.

[0084] In this embodiment, a liquid level sensor 38 , which is optionally coupled to a controller 39 , is included and disposed within the processing vessel to measure the liquid level of the processing vessel.

[0085] In this embodiment, the system is partially or completely composed of single-use components, which can be adapted to be sterilized, for example by gamma irradiation and / or autoclaving, and then connected together using sterile connections, for example of the CPC AseptiQuik type. The alternating flow pump 31 can be single-use or reusable, and it can even be assembled from the following parts: a single-use part, which includes the pump chamber and the mover; and a reusable part, which includes the drive. The single-use part of the alternating flow pump 31 can also be integrated with the filter unit in this case.

[0086] In this embodiment, although the drawings show that the alternating flow pump 31 is placed below the first and second filters, other orientations are also possible, for example by placing the alternating flow pump 31 above the filters. The specific orientation may depend, for example, on space and distance considerations, as well as the use of hydrostatic pressure, for different purposes. Example

[0087] Combination Fig.13 As shown, this embodiment 2 provides a fluid filtration system method based on the embodiment 1, that is, a method for perfusion culture of cells. This embodiment 2 includes the technical content disclosed in the embodiment 1, and the technical content of this embodiment 2 that is the same as that of the embodiment 1 is not repeated. The differences between this embodiment 2 and the embodiment 1 are described below.

[0088] This embodiment includes the following steps: S1, adding cell culture medium and cells to the bioreactor; S2. culturing cells in the bioreactor; S3, during the culture period, operating the alternating flow pump 31, comprising: S31, applying positive pressure to the first end of the first filter unit 1, discharging the fluid from the pump chamber to the bioreactor chamber via the second end of the first filter unit 1 and the pipe section, during which a portion of the fluid is guided to enter the permeate chamber of the first filter unit 1 via the semi-permeable wall portion of the first filter unit 1; at the same time, applying negative pressure to the first end of the second filter unit 2, sucking the fluid from the bioreactor chamber to the pump chamber via the pipe section and the second end of the second filter unit 2, during which a portion of the fluid is guided to enter the permeate chamber via the semi-permeable wall portion of the second filter unit 2; S32, and reverse the direction of the force applied by the alternating flow pump 31, so that the second end of the first filter unit 1 draws fluid from the bioreactor chamber via the pipe segment, and at the same time, the second end of the second filter unit 2 discharges fluid to the bioreactor chamber via the pipe segment.

[0089] S33, repeating steps S31 and S32 at least once, wherein the permeate chamber, the reactor chamber and the alternating flow pump 31 chamber are part of the same closed filtration system; S4. Aspirating permeate from the permeate outlet port.

[0090] In some embodiments, the method includes adding at least one fluid (e.g., cell culture medium) to the bioreactor during the culture period. This has the advantages that the culture fluid removed via the filter unit can be replenished, and fresh nutrients and other reagents can be supplied to the culture.

[0091] In certain embodiments, the filter unit comprises a microfiltration membrane and the permeate comprises a target substance expressed by the cells. This allows for continuous acquisition of the target substance, which may be a protein such as an immunoglobulin.

[0092] In some embodiments, the filter unit comprises an ultrafiltration membrane, and the target substance expressed by the cells remains in the retentate and is fed back to the bioreactor. This allows for the continuous removal of toxic or other undesirable low molecular weight waste products / metabolites from the cell culture, while the target substance (e.g., immunoglobulin protein) can be obtained from the culture at the end of the culture.

[0093] In certain embodiments, the method includes controlling at least one of the alternating flow pump 31 and the inlet control valve to maintain the pressure read from the inlet pressure sensor 36 during the stroke of the alternating flow pump 31 within a predetermined range. The control can be achieved manually, for example by manual adjustment of the valve according to the pressure gauge reading, or by the controller 39 according to the signal received from the pressure sensor 36. The advantage of this method is that the backwashing of the filter unit caused by the negative pressure generated by the stroke can be controlled and optimized to obtain effective cleaning of the membrane and optionally minimal dilution of the retentate with the backwash permeate. It is also possible to control the pressure according to a more complex algorithm, for example, to produce a desired pressure-to-time profile during the stroke.

[0094] In some embodiments, the method includes: calculating the transmembrane pressure based on data provided by the first and second end pressure sensors 36 or from at least one of the first end and pressure sensors 36 and the permeate pressure sensor 36; and controlling at least one of the alternating flow pump 31, the permeate pump 17, and the first end control valve to maintain the transmembrane pressure between preset upper and lower limits. This has the following advantages: the mass transported across the filter membrane can be maximized, and the fouling and concentration polarization can be minimized. It is also possible to control the pressure according to more complex algorithms, such as producing a desired transmembrane pressure-to-time profile during both the suction stroke and the discharge stroke.

[0095] In certain embodiments, the method includes operating the permeate pump 17 in the backward direction to produce a backwash of the filter membrane. The permeate pump may be the sole source of backwash, or the permeate pump may thus be operated during the suction stroke of the alternating flow pump 31 to facilitate backwashing and produce a desired pressure / flow profile during the backwash cycle. The permeate pump may be controlled by one or more controllers 39 based on input data from, for example, an osmotic pressure sensor 36.

[0096] When the system consists of single use components, it is economically feasible to discard all components after use.The system described above is particularly suitable for single use applications, but it is also possible to use a conventional alternating tangential flow system in single use mode. Example

[0097] like Fig.14 As shown, the third embodiment is further modified on the basis of the first embodiment, mainly a mode B system with alternating tangential flow with backwashing. The third embodiment includes the technical contents disclosed in the first embodiment, and the technical contents of the third embodiment that are the same as those of the first embodiment are not repeated. The differences between the third embodiment and the first embodiment are described below.

[0098] This embodiment provides a duplex mode fluid filtration system, which uses a magnetically driven mover to perform two-phase pumping in a sterile sealed container to create an alternating tangential flow with periodic backwashing in the filter unit (mode B system). The system includes: Handling containers; a first filter unit 1 having a first end and a second end and being in fluid communication with the processing vessel; a second filter unit 2 having a first end and a second end and being in fluid communication with the processing vessel; a driver comprising a magnetic field, and An alternating flow pump 31 is connected to the first end of the first filter unit 1 and the second filter unit 2, and has a mover completely sealed in a pump chamber, and the mover has a device that can generate a magnetic field, which can be coupled with the magnetic field of the driver and driven by the change of the driver magnetic field 7. The alternating flow pump 31 can be operated to pump the fluid from the processing container through the first filter unit 1 and the second filter unit 2.

[0099] In some embodiments, Figure 8 As shown, the first filter and the second filter are respectively connected to the pipeline branch point through the first permeate pipeline and the second permeate pipeline, and the pipeline branch point is further connected to the permeate pump 17. During the operation of the alternating flow pump 31, an alternating tangential flow of the fluid (culture solution) is generated inside the first filter and the second filter, and the connected permeate pipelines generate an alternating flow of the permeate, and part of the permeate is returned from the first and second permeate pipelines through the permeate chamber of the filter to the retentate chamber of the filter, forming a backwash of the filter membrane.

[0100] During the α stroke of the alternating flow pump 31, the mover moves toward the first end away from the first filter, generating negative pressure on the retentate side of the first filter, and the first filter is in a suction stroke (negative pressure), so that a certain amount of permeate backwashes the first filter membrane and thereby cleans the cells and other materials that clog the membrane, and the fluid (culture fluid) is sucked from the processing container (bioreactor) through the retentate chamber of the first filter via the pipeline section and enters the first pump chamber 5 of the alternating flow pump 31; at the same time, the second filter is in a discharge stroke (positive pressure), and the fluid (culture fluid) is pushed through the retentate side of the second filter, thereby generating pumpable permeate, and the fluid (culture fluid) further returns to the processing container (bioreactor) via the retentate chamber of the second filter and the pipeline section.

[0101] During the β stroke of the alternating flow pump 31, a force opposite to that during the α stroke is applied to move the mover in the direction away from the first end of the second filter, and the direction of the fluid passing through the first filter and the second filter is opposite to that during the α stroke, that is, when the second filter is in the suction stroke (negative pressure), the fluid (culture fluid) is sucked from the processing container (bioreactor) through the retentate chamber of the second filter into the second pump chamber 10 of the alternating flow pump 31, and the permeate is sucked into the retentate chamber of the second filter through the second permeate pipeline, forming backwashing; at the same time, when the first filter is in the discharge stroke (positive pressure), the fluid (culture fluid) is discharged to the processing container (bioreactor) through the retentate chamber of the first filter, and the fluid (culture fluid) is pushed through the retentate side of the second filter, forming permeation; The flow route of this mode B system allows the negative pressure to be controlled during the suction stroke (negative pressure) of the first and / or second filters to avoid bubble formation and optimize backwashing to make it uniform and well-controlled, especially for cell perfusion culture with higher density and / or longer cycle, which can further reduce the sensitivity of the filtration system to membrane blockage and fouling, increase the filter life, reduce the number of filters used, and thus reduce production costs. Example

[0102] like Fig.15 As shown, the fourth embodiment is further modified on the basis of the first embodiment, mainly a mode C system with a tangential flow with backwashing. The fourth embodiment includes the technical contents disclosed in the first embodiment, and the technical contents of the fourth embodiment and the first embodiment are not repeated here. The differences between the fourth embodiment and the first embodiment are described below.

[0103] In a fourth aspect of the present invention, a duplex mode fluid filtration system is disclosed, which uses a magnetically driven mover to perform two-phase pumping in a sterile sealed container to create a tangential flow with periodic backwashing in the filter unit (mode C system). The system includes: Handling containers; a first filter unit 1 having a first end and a second end and being in fluid communication with the processing vessel; a second filter unit 2 having a first end and a second end and being in fluid communication with the processing vessel; a driver comprising a magnetic field, and An alternating flow pump 31 is connected to the first end of the first filter unit 1 and the second filter unit 2, and has a mover completely sealed in a pump chamber, and the mover has a device that can generate a magnetic field, which can be coupled with the magnetic field of the driver and driven by the change of the driver magnetic field 7. The alternating flow pump 31 can be operated to pump the fluid from the processing container through the first filter unit 1 and the second filter unit 2.

[0104] In some embodiments, as shown, the first end of the first filter is connected to the processing container (bioreactor) via a first end check valve, the first end check valve is arranged to allow flow in the direction from the processing container (bioreactor) to the first end and hinder flow in the opposite direction; and the second end of the first filter is connected to the processing container (bioreactor) via a second end check valve, the second end check valve is arranged to allow flow in the direction from the second end to the processing container (bioreactor) and hinder flow in the opposite direction; In some embodiments, a first end of the second filter is fluidly connected to the processing vessel via a first end check valve; and a second end of the second filter is connected to the processing vessel (bioreactor) via a second end check valve.

[0105] In some embodiments, the first end of the first filter and the second end check valve are each fluidly connected to a pipe branch point, which is further connected to a processing vessel (bioreactor) via a pipe segment.

[0106] In some embodiments, the first end of the second filter and the second end check valve are each fluidly connected to a pipe branch point, which is further connected to a processing vessel (bioreactor) via a pipe segment.

[0107] In some embodiments, optionally, a first end control valve between the first end of the first filter and the branch point, and a second end control valve between the second end of the first filter and the branch point are further included.

[0108] In some embodiments, optionally, a first end control valve between the first end of the second filter unit 2 and the branch point, and a second end control valve between the second end and the branch point are further included.

[0109] In some embodiments, the first filter and the second filter are each connected to a pipe branch point through a first permeate pipe and a second permeate pipe, and the pipe branch point is further connected to a permeate pump 17 .

[0110] During the operation of the alternating flow pump 31, tangential flow of fluid (culture solution) is generated inside the first filter and the second filter, and the connected permeate pipelines generate alternating flow of permeate, and part of the permeate is returned from the first and second permeate pipelines through the permeate chamber of the filter to the retentate chamber of the filter, forming backwashing of the filter membrane.

[0111] During the α stroke of the alternating flow pump 31, the mover moves toward the first end away from the first filter, generating negative pressure on the retentate side of the first filter, and the first filter is in a suction stroke (negative pressure), so that a certain amount of permeate backwashes the first filter membrane and thereby cleans the cells and other materials that clog the membrane, and the fluid (culture fluid) is sucked from the processing container (bioreactor) to the first pump chamber 5 of the alternating flow pump 31 via the pipeline segment; at the same time, the second filter is in a discharge stroke (positive pressure), and the fluid (culture fluid) is pushed through the retentate side of the second filter, thereby generating pumpable permeate, and the fluid (culture fluid) further returns to the processing container (bioreactor) via the retentate chamber and pipeline segment of the second filter.

[0112] During the β stroke of the alternating flow pump 31, a force opposite to that during the α stroke is applied to move the mover in the direction away from the first end of the second filter, and the direction of the fluid passing through the first filter and the second filter is opposite to that during the α stroke, that is, when the second filter is in the suction stroke (negative pressure), the fluid (culture fluid) is sucked from the processing container (bioreactor) into the second pump chamber 10 of the alternating flow pump 31, and the permeate is sucked into the retentate chamber of the second filter through the second permeate pipeline, forming backwashing; at the same time, when the first filter is in the discharge stroke (positive pressure), the fluid (culture fluid) is discharged into the processing container (bioreactor) through the retentate chamber of the first filter, and the fluid (culture fluid) is pushed through the retentate side of the second filter, forming permeation; The flow route of this mode C system allows the negative pressure to be controlled during the suction period (negative pressure) of the first and / or second filters to avoid bubble formation and optimize the backwashing to make it uniform and well controlled, especially for cell perfusion culture with higher density and / or longer cycle, which can further reduce the sensitivity of the filtration system to membrane blockage and fouling, increase the filter life, reduce the number of filters used, and thus reduce production costs. Example

[0113] like Fig.16 As shown, the fifth embodiment is further modified on the basis of the first embodiment, mainly to expand the pump system, and the modes ABC are compatible. The fifth embodiment includes the technical contents disclosed in the first embodiment, and the technical contents of the fifth embodiment and the first embodiment are not repeated here. The differences between the fifth embodiment and the first embodiment are described below.

[0114] The present embodiment provides a duplex mode fluid filtration system, which uses multiple alternating flow pumps 31 to create alternating tangential flow in the filter unit, and optionally create alternating tangential flow with backwashing in the filter unit, and optionally create tangential flow with backwashing in the filter unit. This system is compatible with the mode A, B, and C systems.

[0115] The system includes: Handling containers; a first filter unit 1 having a first end and a second end and being in fluid communication with the processing vessel; a second filter unit 2 having a first end and a second end and being in fluid communication with the processing vessel; a main drive, the main drive comprising a magnetic field; a main alternating flow pump 31 connected to the first ends of the first filter unit 1 and the second filter unit 2; and at least one extension drive, the extension drive comprising a magnetic field; At least one extended alternating flow pump 31 is connected to the first ends of the first filter unit 1 and the second filter unit 2 through a manifold.

[0116] In some embodiments, the α stroke and β stroke of the main pump and the extension pump can be synchronized. Optionally, the extension pump can be different in size from the main pump and can be controlled by the same controller 39 as the main pump, or alternatively, can be controlled by a different controller 39.

[0117] The extended pump arrangement has the following features, which are very important for perfusion culture systems that are scaled up to 2000L and above: The extension pump can be used to reduce the stroke length of the main pump, reducing the space requirements in the GMP clean room, thereby increasing the flexibility and economy of the layout of the bioreactor or filtration system; Optionally, the extension pump can be used to moderate the movement speed of the main pump, which can reduce the fluid shear force in the pump chamber, thereby reducing the effect of the fluid shear force on fragile materials in the fluid, such as animal cells, biomolecules, etc.; Optionally, an extension pump can be used to reduce the diameter of the main pump, which not only reduces the strength requirement of the magnetic coupling and avoids the risk of demagnetization, but also reduces the friction area and thus reduces the fluid heating phenomenon caused by the friction effect, thereby reducing the impact on fragile materials in the fluid such as animal cells, biomolecules, etc.

[0118] In some embodiments, optionally, the cycle and phase of the extension pump can be different from that of the main pump, including being opposite to the phase of the main pump (the α stroke of the main pump is synchronized with the β stroke of the extension pump, and vice versa, that is, the β stroke of the main pump can be synchronized with the α stroke of the gain pump) and remaining stationary during the operation of the main pump, etc. This allows the chambers of the main pump and the extension pump to be used for fluid mixing, fluid sampling, and delivering bubbles in the fluid to a designed bubble trap.

[0119] In some embodiments, the expansion pump optionally provides a larger displacement and can be used to enhance the alternating tangential flow of the system in Mode A, optionally enhance the alternating tangential flow and backwash of the system in Mode B, and optionally enhance the tangential flow and backwash of the system in Mode C. The expansion pump can be programmed to perform a perfusion procedure or / and to perform a filter cleaning procedure. Example

[0120] like Fig.17 As shown, the sixth embodiment is further modified on the basis of the first embodiment, mainly to expand the pump system, and the modes ABC are compatible. The sixth embodiment includes the technical contents disclosed in the first embodiment, and the technical contents of the sixth embodiment and the first embodiment are not repeated here. The differences between the sixth embodiment and the first embodiment are described below.

[0121] The present embodiment provides a duplex mode fluid filtration system, which uses multiple alternating flow pumps 31 to create alternating tangential flow in the filter unit, and optionally create alternating tangential flow with backwashing in the filter unit, and optionally create tangential flow with backwashing in the filter unit. This system is compatible with the mode A, B, and C systems.

[0122] The system includes: Handling containers; a first filter unit 1 having a first end and a second end and being in fluid communication with the processing vessel; a second filter unit 2 having a first end and a second end and being in fluid communication with the processing vessel; a main drive, the main drive comprising a magnetic field; a main alternating flow pump 31 connected to the first ends of the first filter unit 1 and the second filter unit 2; and at least one gain driver, the gain driver comprising a magnetic field; At least one gain alternating flow pump 31 is connected to the second ends of the first filter unit 1 and the second filter unit 2 .

[0123] In some embodiments, the alpha stroke of the main pump can be synchronized with the beta stroke of the gain pump, and vice versa, that is, the beta stroke of the main pump can be synchronized with the alpha stroke of the gain pump. Optionally, the gain pump size can be different from the main pump, and it can be controlled by the same controller 39 as the main pump, or alternatively, it can be controlled by a different controller 39.

[0124] The gain pump arrangement has the following features, which are very important for perfusion culture systems that are scaled up to 2000 L and above: The boost pump may provide an improvement in the scalability of the system as it is able to keep the pressure and differential pressure across the membrane constant regardless of the tubing diameter and length as well as the pressure inside the bioreactor and the hydrostatic pressure caused by the vertical distance between the bioreactor and the filter unit.

[0125] The boost pump can overcome the effects of gravity and pressure decay within the filter, providing a more uniform flow, thereby providing the ability to more uniformly utilize the filter membrane area and / or hollow fiber tube length.

[0126] In some embodiments, the alpha stroke and beta stroke of the main pump and the boost pump may optionally be synchronized.

[0127] In some embodiments, the boost pump may optionally remain stationary during operation of the main pump.

[0128] In some embodiments, optionally, the boost pump provides a larger displacement, and can be used to enhance the alternating tangential flow of the mode A system, optionally, enhance the alternating tangential flow and backwash of the mode B system, optionally, enhance the tangential flow and backwash of the mode C system, and optionally, create the alternating tangential flow and backwash of the mode C system when the control valve of the pipeline connecting the process container (reactor) is opened or closed. The boost pump can be programmed to perform a perfusion program or / and perform a filter cleaning program.

[0129] In some embodiments, optionally, the gain pump can be used to reduce the stroke length of the main pump, thereby reducing the space requirements in the GMP clean room, thereby increasing the flexibility and economy of the layout of the bioreactor, i.e., the filtration system; optionally, the gain pump can be used to moderate the movement speed of the main pump, which can reduce the fluid shear force in the pump chamber, thereby reducing the effect of the fluid shear force on fragile materials in the fluid, such as animal cells, biological molecules, etc.; optionally, the gain pump can be used to reduce the diameter of the main pump, which can not only reduce the strength requirements of the magnetic coupling and avoid the risk of demagnetization, but also reduce the friction area to reduce the fluid heating phenomenon caused by the friction effect, thereby reducing the effect on fragile materials in the fluid such as animal cells, biological molecules, etc.

[0130] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A filtration system, characterized in that: include: Handling containers; a first filter unit having a first end and a second end and being in fluid communication with the processing vessel; a second filter unit having a first end and a second end and being in fluid communication with the processing vessel; A driver for generating a magnetic field; An alternating flow pump is connected to the first ends of the first filter unit and the second filter unit, and has a mover that is completely sealed in a pump chamber. The mover can generate a magnetic field that couples with the magnetic field of the driver. The driver drives the mover to reciprocate in the pump chamber by controlling changes in the magnetic field, so as to pump the fluid from the processing container through the first filter unit and the second filter unit.

2. The filtration system according to claim 1, characterized in that: The alternating flow pump generates an alternating tangential flow of the fluid inside the first filter unit and the second filter unit.

3. The filtration system according to claim 1, characterized in that: The mover has a sealing device, which divides the pump chamber of the alternating flow pump into a first chamber and a second chamber; when the mover moves toward the first end direction away from the first filter unit, that is, the α stroke, the alternating flow pump sucks the fluid in the first filter unit and the processing container into the first chamber of the alternating flow pump; at the same time, the fluid in the second chamber of the alternating flow pump is discharged into the second filter unit and the processing container; when the mover moves toward the first end direction away from the second filter unit, that is, the β stroke, the alternating flow pump sucks the fluid in the second filter unit and the processing container into the second chamber of the alternating flow pump; at the same time, the fluid in the first chamber of the alternating flow pump is discharged into the first filter unit and the processing container.

4. The filtration system according to claim 1, characterized in that: The coupling between the magnetic field of the mover of the alternating current pump and the magnetic field of the driver includes concentric coupling, end surface coupling, or a composite coupling.

5. The filtration system according to claim 1, characterized in that: The change of the driver magnetic field includes magnetic field flux change, magnetic field displacement, and magnetic pole conversion.

6. The filtration system according to claim 3, characterized in that: The sealing device is a liquid sealing device arranged on the mover.

7. The filtration system according to claim 1, characterized in that: Also includes: A controller is connected to the driver and is used to control the movement of the driver to controllably move the mover.

8. The filtration system according to claim 1, characterized in that: Also includes: a first end pressure sensor arranged to measure pressure between first ends of the first and second filter units and the alternating flow pump chamber; and / or, a second end pressure sensor arranged to measure pressure between the second ends of the first and second filter units and the process vessel; The controller is arranged to control the driver to control the alternating flow pump based on pressure data received from at least one of the first end and second end pressure sensors of the first filter.

9. The filtration system according to claim 1, characterized in that: Also includes: A linear encoder is connected to the drive for determining the position of the movable drive.

10. The filtration system according to any one of claims 1 to 9, characterized in that: The first filter unit and the second filter unit are each connected to a pipeline branch point through a first permeate pipeline and a second permeate pipeline, and the pipeline branch point is connected to a permeate pump.

11. The filtration system according to claim 10, characterized in that: The alternating flow pump generates alternating flow in the first permeate pipe and the second permeate pipe, and part of the permeate is returned from the first permeate pipe and the second permeate pipe to the retentate chamber of the filter through the permeate chamber of the filter, thereby backwashing the filter membrane.

12. The filtration system according to claim 10, characterized in that: When the mover moves in the direction away from the first end of the first filter unit, i.e., the α stroke, the alternating flow pump sucks the fluid in the first filter unit and the processing container into the first chamber of the alternating flow pump, and the fluid passing through the first permeate pipeline returns to the retentate chamber of the first filter, forming a backwash of the filter membrane; at the same time, the fluid in the second chamber of the alternating flow pump is discharged into the second filter unit and the processing container, and forms a permeate; When the mover moves in the direction away from the first end of the second filter unit, that is, the β stroke, the alternating flow pump sucks the fluid in the second filter unit and the processing container into the second chamber of the alternating flow pump, and the fluid through the second permeate pipeline returns to the retentate chamber of the second filter, forming backwashing of the filter membrane; at the same time, the fluid in the first chamber of the alternating flow pump is discharged into the first filter unit and the processing container, and permeation is formed.

13. The filtration system according to any one of claims 1 to 9, characterized in that: The first end of the first filter unit is connected to the processing container via a first end check valve, and the second end of the first filter unit is connected to the processing container via a second end check valve; the first end of the second filter unit is connected to the processing container via the first end check valve; the second end of the second filter unit is fluidly connected to the processing container via the second end check valve; the first end check valve is arranged to allow fluid flow in a direction from the processing container to the first end and hinder flow in the opposite direction; the second end check valve is arranged to allow fluid flow in a direction from the second end to the processing container and hinder flow in the opposite direction.

14. The filtration system according to claim 13, characterized in that: Also includes: The first end check valve and the second end check valve of the first filter unit are respectively connected to a pipeline branch point, and the pipeline branch point is further connected to the processing container via a pipeline section; a first end control valve is arranged between the first end of the first filter unit and the pipeline branch point, and a second end control valve is arranged between the second end of the first filter unit and the branch point.

15. The filtration system according to claim 13, characterized in that: Also includes: The first end check valve and the second end check valve of the second filter unit are respectively connected to a pipeline branch point, and the pipeline branch point is further connected to the processing container via a pipeline section; a first end control valve is arranged between the first end of the second filter unit and the pipeline branch point, and a second end control valve is arranged between the second end of the second filter unit and the pipeline branch point.

16. The filtration system according to any one of claims 1 to 9, characterized in that: The alternating flow pump comprises: a main alternating flow pump connected to the first end of the first filter unit and the second filter unit; at least one extended alternating flow pump connected to the first end of the first filter unit and the second filter unit through a manifold; The driver comprises: A main driver, the main driver being adapted to the main alternating flow pump; At least one expansion drive adapted to the expansion alternating flow pump.

17. The filtration system according to any one of claims 1 to 9, characterized in that: The alternating flow pump comprises: a main alternating flow pump connected to the first end of the first filter unit and the second filter unit; at least one gain alternating flow pump connected to the first filter unit and the second end of the second filter unit; The driver comprises: A main driver, the main driver being adapted to the main alternating flow pump; At least one gain driver, the gain driver is adapted to the gain alternating current pump.

18. A filtering method according to any one of claims 1 to 17, characterized in that: The steps include: S1, adding cell culture medium and cells to the bioreactor; S2. culturing cells in the bioreactor; S3. During the culture period, operating the alternating flow pump, comprising: S31, applying positive pressure to the first end of the first filter unit, discharging the fluid from the pump chamber to the bioreactor chamber via the second end of the first filter unit and the pipe section, during which a portion of the fluid is guided to enter the permeate chamber of the first filter unit via the semi-permeable wall portion of the first filter unit; and simultaneously, applying negative pressure to the first end of the second filter unit, sucking the fluid from the bioreactor chamber to the pump chamber via the pipe section and the second end of the second filter unit, during which a portion of the fluid is guided to enter the permeate chamber via the semi-permeable wall portion of the second filter unit; S32, and reversing the direction of the force applied by the alternating flow pump, so that the second end of the first filter unit draws fluid from the bioreactor chamber via the pipe segment, and simultaneously the second end of the second filter unit discharges fluid to the bioreactor chamber via the pipe segment; S33, repeating steps S31 and S32 at least once, wherein the permeate chamber, the reactor chamber and the alternating flow pump chamber are part of the same closed filtration system; S4. Aspirating permeate from the permeate outlet port.

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