A cell culture fluid clarification harvest system

Abstract

The cell culture liquid clarification harvesting system provided by the utility model, through the pressure regulating device arranged between the secondary membrane and the feed sterilization filter for adjusting the fluid flow entering the discharge sterilization filter, offsetting the pressure superposition influence of the feed sterilization filter inlet pressure and the pressure rise on the primary membrane inlet, so that the cell integrity is prevented from being damaged in the filtering process caused by the excessively high primary membrane inlet pressure; the venting system is arranged at the bottom of the harvesting tank, the venting pipeline and the sampling pipeline are communicated with the harvesting tank by the L-shaped pipeline, the on-line sampling for the oxygen partial pressure detection of the harvesting liquid can be realized, the oxygen supply amount of the venting system can be automatically adjusted according to the oxygen partial pressure data, so that the problem that the sampling cannot be realized and the oxygen partial pressure of the harvesting liquid cannot be monitored at the low liquid level is effectively avoided, the problem that the harvesting liquid cannot be vented at the low liquid level is solved, and the purpose of adjustable oxygen partial pressure is achieved.

Description

Technical Field

[0001] This utility model relates to a cell culture medium clarification and harvesting system. Background Technology

[0002] Clarification harvesting systems are commonly used equipment for filtering supernatants after cell culture / fermentation in biopharmaceuticals, health products, and other products.

[0003] Clarification filtration is a common method for filtering cell supernatant after cell culture. It involves using a primary membrane to filter out insoluble large particulate impurities from the culture medium, a secondary membrane to filter out bacteria and small insoluble particulate impurities, and finally using a liquid sterile filter cartridge to filter out microorganisms, in order to obtain sterile supernatant.

[0004] The clarification and harvesting system filtration in a GMP production workshop typically consists of a bioreactor, a power pump, a primary membrane, a secondary membrane, a feed sterile filter, a harvesting tank, an outlet sterile filter, and related sensors. During operation, the reactor's outlet is connected to the power pump via piping. The power pump's downstream end is sequentially connected to the primary membrane (in parallel), the secondary membrane (in parallel), the feed sterile filter, the harvesting tank, and the outlet sterile filter, before entering the chromatography stage. At startup, the power pump draws the culture medium through the primary membrane stack at the process flow rate to remove cells and large insoluble particles. It then flows through the secondary membrane to remove bacteria and small insoluble particles. The medium is then temporarily stored in the sterile harvesting tank via the feed sterile filter. Before subsequent chromatography, the medium passes through the outlet sterile filter; two sterile filters are installed to ensure the harvested medium temporarily stored in the harvesting tank remains relatively sterile.

[0005] During operation, the aforementioned conventional clarification and harvesting system not only frequently suffers from excessively high inlet pressure at the primary membrane, which can disrupt cell integrity during filtration, causing contents to precipitate out and release substances that damage the target protein, thus affecting its molecular weight properties; but also, when the target protein is temporarily stored in the harvesting tank, fluctuations in dissolved oxygen levels can lead to its reduction, further impacting its quality properties.

[0006] Therefore, during the conventional clarification and harvesting process of cell culture medium, if filtration is performed at a predetermined process flow rate, the primary membrane inlet pressure cannot be maintained within a specific range, leading to production halt. Furthermore, since the clarification and harvesting system lacks a ventilation control system, although the bioreactor ventilation system can currently be used to aerate the harvested liquid, if the liquid level is below the vent, ventilation and sampling of the harvested liquid will be impossible. This makes it impossible to monitor the oxygen partial pressure during storage to maintain dissolved oxygen balance within the harvesting tank. Consequently, the target protein temporarily stored in the harvesting tank may be reduced due to fluctuations in the dissolved oxygen environment, affecting the quality properties of the target protein.

[0007] Existing technologies use a reduced clarification harvest flow rate during the pilot phase to control the primary membrane pressure and keep it within a low range. However, this means that the process parameters cannot be matched with GMP-scale production, which will lead to a series of subsequent problems in commercial production.

[0008] Therefore, there is a need for a cell culture medium clarification and harvesting system that can solve the above-mentioned technical problems. Utility Model Content

[0009] To address the aforementioned problems, this invention provides a cell culture clarification and harvesting system that can effectively control both the primary membrane inlet pressure and the oxygen partial pressure inside the harvesting tank.

[0010] The present invention provides a cell culture medium clarification and harvesting system, which mainly consists of a bioreactor, a power pump, a primary membrane, a secondary membrane, a feed sterilization filter, and a harvesting tank. The system is characterized in that a pressure regulating device is provided between the secondary membrane and the feed sterilization filter to regulate the pressure at the inlet of the primary membrane.

[0011] In some embodiments, the pressure regulating device is a buffer system configured to regulate the pressure at the primary membrane inlet by controlling the amount of fluid flowing through the buffer system, thereby offsetting the cumulative effect of the rising pressure generated by the fluid in the pipeline between the feed sterilizer and the harvest tank on the pressure at the primary membrane inlet, and thus avoiding the destruction of cell integrity during the filtration process due to excessively high pressure at the primary membrane inlet.

[0012] In some embodiments, the buffer system is a buffer bag configured to buffer the fluid delivered through the secondary membrane outlet. The amount of fluid buffered is adjusted by its own volume change or a peristaltic pump to offset the pressure superposition effect of the rising pressure generated by the fluid in the pipeline between the feed sterilization filter and the harvest tank on the primary membrane inlet.

[0013] In some specific embodiments, the cushioning bag is a flexible bag made of an elastic material, wherein the elastic material may be selected from any one or more of medical silicone, bio-based isoprene rubber, bio-based EPDM rubber, polyurethane, or PET polyester.

[0014] In some specific embodiments, the buffer bag is provided with a feed tube, the proximal end of which is connected to the outlet of the secondary membrane, and the distal end extends into the bag.

[0015] In some specific embodiments, the buffer system is further provided with a peristaltic pump at the outlet of the buffer bag. The peristaltic pump is configured to pump the buffer fluid in the buffer bag into the feed sterilization filter. When the amount of fluid in the buffer bag exceeds a threshold, the pumping speed of the peristaltic pump is adjusted by PID so that the fluid in the buffer bag can quickly enter the feed sterilization filter, offsetting the pressure superposition effect of the rising pressure generated by the fluid in the pipeline between the feed sterilization filter and the harvest tank on the primary membrane inlet.

[0016] In some specific embodiments, a weighing sensor is also provided at the bottom of the buffer bag. The weighing sensor transmits the fluid volume signal in the buffer bag to the PID. The PID automatically sets the pump speed of the peristaltic pump according to the fluid volume and automatically adjusts the fluid volume entering the feed sterilization filter to offset the pressure superposition effect of the rising pressure generated by the fluid in the pipeline between the feed sterilization filter and the harvest tank on the primary membrane inlet.

[0017] In some embodiments, the buffer system is a diaphragm pump configured to automatically adjust the pump speed of the diaphragm pump according to the pressure at the outlet of the secondary membrane, thereby controlling the amount of fluid entering the feed sterilizer and offsetting the cumulative effect of the rising pressure generated by the fluid in the pipeline between the feed sterilizer and the harvest tank on the pressure at the inlet of the primary membrane.

[0018] In some embodiments, the cell culture medium clarification harvesting system is equipped with a ventilation system at the bottom of the harvesting tank. The ventilation system mainly consists of an L-shaped pipe, a three-way valve, a ventilation pipe, and a sampling pipe installed on the harvesting tank.

[0019] The distal end of the L-shaped pipeline extends into the harvest tank, and the proximal end is connected to the ventilation pipeline and the sampling pipeline via a three-way valve. The three-way valve controls the connection and disconnection of the ventilation pipeline and the sampling pipeline with the harvest tank to achieve ventilation or oxygen partial pressure sampling and detection in the harvest tank.

[0020] In some specific embodiments, the ventilation line is provided with a gas flow controller (MFC) configured to control the intake of oxygen or air.

[0021] In some specific embodiments, the angle formed between the distal and proximal ends of the L-shaped conduit is greater than 80°.

[0022] In some specific embodiments, the extension of the L-shaped pipe into the harvesting tank is perpendicular to the liquid surface inside the harvesting tank.

[0023] In some specific embodiments, the distal end of the L-shaped pipe extending into the harvesting tank extends to the bottom of the tank body, thereby ensuring that the harvesting tank can sample and ventilate the harvested liquid throughout the entire process of harvesting and temporarily storing the harvested liquid, so as to maintain the oxygen partial pressure value in the harvested liquid tank within the process requirements range.

[0024] In some specific embodiments, a gas flow controller (MFC) may also be provided on the ventilation line, the gas flow controller (MFC) being configured to adjust the oxygen content of the gas supplied to the harvest tank in real time based on the partial pressure of oxygen in the harvest liquid within the harvest tank.

[0025] In some specific embodiments, an oxygen content sensor is provided inside the harvesting tank. The oxygen content sensor is configured to be electrically connected to the gas flow controller (MFC) to detect the oxygen partial pressure of the harvested liquid in the harvesting tank in real time and transmit the oxygen partial pressure detection data to the gas flow controller (MFC), so that the gas flow controller (MFC) can automatically regulate the amount of oxygen input in the ventilation pipeline.

[0026] In some specific embodiments, the oxygen content sensor may be selected from a zirconia oxygen sensor, an optical fluorescence quenching sensor, or an electrochemical sensor, preferably an electrochemical sensor.

[0027] The secondary membrane inlet pressure, the feed sterilizer inlet pressure, and the ramp pressure (H) generated by the feed sterilizer and the liquid inlet of the harvest tank can be detected in real time by a pressure gauge.

[0028] Compared with the prior art, the beneficial effects of this utility model are as follows:

[0029] 1. The cell culture medium clarification and harvesting system of this utility model has a pressure regulation device between the secondary membrane and the feed sterilization filter. By regulating the amount of fluid flowing through the buffer system, the pressure rise generated by the fluid in the pipeline between the feed sterilization filter and the harvesting tank is offset from the pressure superimposed on the primary membrane inlet. This avoids the cell integrity being damaged during the filtration process due to excessively high pressure at the primary membrane inlet, reduces the pressure on individual cells, and simultaneously meets the process requirements of flow rate and pressure.

[0030] 2. The cell culture medium clarification and harvesting system of this utility model has an aeration system at the bottom of the harvesting tank. An L-shaped pipeline connects the aeration pipeline and the sampling pipeline to the harvesting tank. This system can realize online detection of the oxygen partial pressure of the harvested liquid and automatically adjust the oxygen supply of the aeration system according to the oxygen partial pressure data to achieve adjustable oxygen partial pressure. It can also effectively avoid the problems of not being able to sample at low liquid levels and aeration of the harvested liquid. Attached Figure Description

[0031] Figure 1 This is a schematic diagram of the cell culture medium clarification and harvesting system with a buffer bag as the pressure regulating device in Embodiment 1 of this application.

[0032] Figure 2 This is a schematic diagram of the specific structure of the buffer bag in Embodiment 1 of this application.

[0033] Figure 3 This is a schematic diagram of the cell culture medium clarification and harvesting system with a diaphragm pump as the pressure regulating device in Embodiment 2 of this application.

[0034] Figure 4 is a schematic diagram of the cell culture medium clarification and harvesting system containing a ventilation system in Embodiment 3 of this application (wherein the pressure regulating device in the schematic diagram of the cell culture medium clarification and harvesting system in 4a is a buffer bag, and the pressure regulating device in the schematic diagram of the cell culture medium clarification and harvesting system in 4b is a diaphragm pump).

[0035] Illustration:

[0036] 1. Bioreactor

[0037] 2. Power pump

[0038] 3. Primary membrane

[0039] 4. Secondary membrane

[0040] 5. Feed sterilization filter

[0041] 6. Harvesting tank

[0042] 7. Pressure regulating device, 71-buffer bag, 711-feed pipe, 72-peristaltic pump, 73-weighing sensor, 74-diaphragm pump

[0043] 8. Ventilation system, 81-L-type piping, 82-Three-way valve, 83-Ventilation piping, 84-Sampling piping, 85-Gas quality controller, 86-Oxygen content sensor Detailed Implementation

[0044] The technical solutions of the utility model embodiments will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the utility model, and not all embodiments. Based on the embodiments of the utility model, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the utility model.

[0045] Example 1

[0046] Please see Figure 1-2The cell culture medium clarification and harvesting system provided in this embodiment mainly includes: a bioreactor 1, a power pump 2, a primary membrane 3, a secondary membrane 4, a feed sterilizing filter 5, and a harvesting tank 6. The outlet of the bioreactor 1 is connected to the inlet pipe of the power pump 2. The outlet pipe of the power pump 2 is sequentially connected to the primary membrane 3, the secondary membrane 4, and the feed sterilizing filter 5. The outlet pipe of the feed sterilizing filter 5 is connected to the inlet of the harvesting tank 6. Thus, the cell culture medium of the bioreactor 1 passes through the primary membrane 3 to remove cells and large insoluble particles, and the secondary membrane 4 to remove bacteria and small insoluble particles, according to the process flow rate provided by the power pump 2. After sterilization and filtration by the feed sterilizing filter 5, the harvested medium is temporarily stored in the harvesting tank 6. The harvested medium temporarily stored in the harvesting tank 6 is then sterilized and filtered by the outlet sterilizing filter before being discharged for subsequent chromatography purification.

[0047] In this embodiment, the cell culture medium clarification and harvesting system is equipped with a pressure regulating device 7 between the secondary membrane 4 and the feed sterilization filter 5. The pressure regulating device 7 is configured to regulate the inlet pressure of the primary membrane 3 to avoid the cell integrity being damaged during the filtration process due to excessively high inlet pressure of the primary membrane 3, resulting in the precipitation of contents and the release of substances that damage the target protein and affect the molecular weight properties of the target protein.

[0048] In this embodiment, the pressure regulating device 7 is equipped with a buffer system 71. The buffer system 71 is configured to regulate the pressure at the inlet of the primary membrane 3 by controlling the amount of fluid flowing through the buffer system 71. This offsets the superimposed effect of the rising pressure generated by the fluid in the pipeline between the feed sterilizer filter 5 and the harvest tank 6 on the pressure at the inlet of the primary membrane 3, thereby preventing the cell integrity from being damaged during the filtration process due to excessively high pressure at the inlet of the primary membrane 3.

[0049] The buffer system is a buffer bag 71, which is configured to buffer the fluid delivered through the outlet of the secondary membrane 4. The amount of fluid in the buffer bag 71 is adjusted by its own volume change or by a peristaltic pump. The pressure at the inlet of the feed sterilization filter 5 and the rising pressure have a superimposed effect on the pressure at the inlet of the primary membrane 3.

[0050] The buffer bag 71 described in this embodiment can be a flexible bag made of an elastic material, wherein the elastic material can be selected from any one or more of medical-grade silicone, bio-based isoprene rubber, bio-based EPDM rubber, polyurethane, or PET polyester. The flexible bag in this embodiment adjusts the fluid buffer volume through its own volume change, and the inlet pressure of the feed sterilization filter 5 and the rising pressure have a combined effect on the inlet pressure of the primary membrane 3.

[0051] In this embodiment, the buffer bag 71 is provided with a feed pipe 711. The proximal end of the feed pipe 711 is connected to the inlet of the secondary membrane 4, and the distal end extends into the bag of the buffer bag 71, thereby preventing air bubbles from being generated during the flow of fluid from the secondary membrane 4 into the buffer bag 71.

[0052] The buffer system described in this embodiment is further provided with a peristaltic pump 72 at the outlet of the buffer bag 71. The peristaltic pump 72 is configured to pump the fluid in the buffer bag 71 into the feed sterilization filter 5. When the volume of the fluid in the buffer bag 71 exceeds the volume threshold of the buffer bag 71, the pumping speed of the peristaltic pump 72 is adjusted by PID so that the fluid in the buffer bag 71 can quickly enter the feed sterilization filter 5, offsetting the combined effect of the inlet pressure of the feed sterilization filter 5 and the pressure of the rising pressure on the inlet of the primary membrane 3.

[0053] The bottom of the buffer bag 71 is also equipped with a weighing sensor 73. The weighing sensor 73 transmits the fluid weight signal in the buffer bag 71 to the PID. The PID automatically sets the pump speed of the peristaltic pump 72 according to the fluid weight and automatically adjusts the amount of fluid entering the feed sterilization filter 5 to offset the combined effect of the inlet pressure of the feed sterilization filter 5 and the pressure superimposed on the inlet of the primary membrane 3 by the rising pressure.

[0054] In this embodiment, the liquid pressure of the pipelines of the primary membrane 3, secondary membrane 4, feed sterilization filter 5, and harvest tank 6 can be tested sequentially using pressure gauges P1, P2, P3, and P4.

[0055] The power pump 2 described in this embodiment can be a diaphragm pump or a peristaltic pump.

[0056] Example 2

[0057] The cell culture medium clarification and harvesting system described in this embodiment differs from that in Example 1 in that, as shown in the attached figure... Figure 3 As shown, the buffer system is a diaphragm pump 74. The diaphragm pump 74 is configured to automatically adjust the pumping speed of the diaphragm pump 74 according to the pressure at the outlet of the primary membrane 3, thereby controlling the amount of fluid entering the feed sterilization filter 5. This offsets the combined effect of the inlet pressure of the feed sterilization filter 5 and the pressure rise on the inlet pressure of the primary membrane 3, thereby preventing the cell integrity from being damaged during the filtration process due to excessively high inlet pressure of the primary membrane 3, resulting in the precipitation of contents and the release of substances that damage the target protein and affect the molecular weight properties of the target protein.

[0058] Example 3

[0059] The cell culture medium clarification and harvesting system provided in this embodiment is based on Embodiment 1 or Embodiment 2, such as... Figure 4a , 4bAs shown, a ventilation system 8 is provided at the bottom of the harvesting tank 6. The ventilation system 8 mainly consists of an L-shaped pipe 81, a three-way valve 82, a ventilation pipe 83, and a sampling pipe 84 installed on the harvesting tank 6.

[0060] The distal end of the L-shaped pipe 81 extends into the harvest tank 6, and the proximal end of the L-shaped pipe 81 is connected to the ventilation pipe 83 and the sampling pipe 84 via a three-way valve 82. The three-way valve 83 controls the connection and disconnection of the ventilation pipe 83 and the sampling pipe 84 with the harvest tank 6 to achieve sampling, ventilation, or oxygen partial pressure detection of the harvest liquid in the harvest tank 6.

[0061] In this embodiment, the ventilation duct 83 is provided with a gas flow controller (MFC) 85, which is configured to control the intake volume of oxygen or air.

[0062] In this embodiment, the angle formed between the distal end and the proximal end of the L-shaped pipe 81 is greater than 90°, preferably 105°, so that the L-shaped pipe 81 can be used to install and fix the harvesting tank 6.

[0063] In this embodiment, the pipe extending into the harvest tank 6 is perpendicular to the liquid surface inside the harvest tank 6, so that when the venting pipe 83 purges the harvest tank 6 to remove residues, it can ensure that there are no residues in the venting system 8 pipe.

[0064] In this embodiment, the distal end of the L-shaped pipe 81 extending into the harvest tank 6 extends to the bottom of the tank body, thereby ensuring that the sampling pipe 84 can sample and ventilate the harvest liquid throughout the entire process of harvesting and temporarily storing the harvest liquid in the harvest tank 6, so as to maintain the oxygen partial pressure value in the harvest liquid tank 6 within the process requirements range.

[0065] In this embodiment, an oxygen content sensor 86 is provided in the harvest tank 6. The oxygen content sensor 86 is configured to be electrically connected to the gas flow controller (MFC) 85. The gas flow controller (MFC) 85 automatically adjusts the amount of oxygen or air input in the ventilation pipeline 83 according to the oxygen partial pressure of the harvest liquid detected by the oxygen content sensor 86 in the harvest tank.

[0066] The oxygen content sensor 86 can be selected from a zirconia oxygen sensor, an optical fluorescence quenching sensor, or an electrochemical sensor, and is preferably an electrochemical sensor.

[0067] The electrochemical sensor described in this invention can be selected from electrochemical dissolved oxygen sensors and optical dissolved oxygen sensors.

[0068] The ventilation system 8 described in this embodiment not only avoids the inability to sample at low liquid levels, thus preventing the monitoring of the oxygen partial pressure of the harvested liquid, but also solves the problem of not being able to ventilate the harvested liquid at low liquid levels, thereby achieving the purpose of adjustable oxygen partial pressure.

[0069] Example 4

[0070] This embodiment utilizes the cell culture medium clarification and harvesting system described in Example 3 to perform cell culture medium clarification and harvesting.

[0071] Equipment used: The cell culture medium clarification and harvesting system described in Example 3, specifically as follows... Figure 4a As shown;

[0072] Protein type: Monoclonal antibody;

[0073] Clarification harvest parameter settings: Filtration rate: 1650 L / h, primary membrane inlet pressure: ≤1 bar, oxygen partial pressure control: 70-110 mmHg, ventilation type: air, ventilation rate: 0.005 vvm

[0074] The specific connection method is as follows: a silicone tube is used to connect the bioreactor 1, power pump 2, primary membrane 3, secondary membrane 4, buffer bag 71, feed sterilizer 5, and harvest tank 6 in one go. The inlet of the buffer bag 71 is connected to the outlet of the secondary membrane 4, and the outlet of the buffer bag 71 is connected to the feed sterilizer 5. The L-shaped pipe 81 of the ventilation system 8 is connected to the ventilation pipe 83 and the sampling pipe 84 respectively through the sterilized three-way valve 82, and is sterilized together with the harvest tank 6 to ensure that the entire cell culture medium clarification and harvesting system has undergone a sterilization process.

[0075] Turn on the power pump 2 and adjust the flow rate settings accordingly. Simultaneously, ensure the inlet pressure of the primary membrane 3 does not exceed 1 bar, and record the readings periodically. The weighing sensor 73 transmits the fluid weight signal from the buffer bag 71 to the PID controller. The PID controller automatically sets the pump speed of the peristaltic pump 72 based on the fluid weight, automatically adjusting the amount of fluid entering the feed sterilizer 5 to offset the combined effect of the inlet pressure of the feed sterilizer 5 and the rising pressure on the inlet pressure of the primary membrane 3.

[0076] When the weighing sensor 73 detects that the weight of the fluid in the buffer bag 71 exceeds the threshold, it automatically sends a signal to the PID controller. The PID controller automatically turns on and adjusts the pump speed of the peristaltic pump 72 according to the weight of the fluid detected by the weighing sensor 73, thereby automatically adjusting the amount of fluid entering the feed sterilization filter 5 to offset the combined effect of the inlet pressure of the feed sterilization filter 5 and the pressure rise on the inlet of the primary membrane 3.

[0077] The fluid weight threshold in the buffer bag 71 is the weight when 1 / 3 of the volume of the buffer bag 71 is used to store fluid. The discharge pump speed of the peristaltic pump 72 is consistent with the pump speed of the power pump 2, thereby maintaining a relatively stable fluid volume in the buffer bag 71 (between 1 / 3 and 2 / 3) to offset the combined effect of the inlet pressure of the feed sterilization filter 5 and the pressure rise on the inlet of the primary membrane 3, until all the cell culture medium is clarified and harvested.

[0078] When the collected liquid in the harvesting tank 6 exceeds 150L, ​​the agitator is activated according to the process requirements. The three-way valve 82 is opened to purge the residual liquid in the L-shaped pipe 81 through the vent pipe 83. Then, the sampling pipe 84 is opened to take a sample. If sampling is not possible due to insufficient pressure in the harvesting tank 6, the pressure in the harvesting tank 6 can be appropriately increased. The above sampling process is repeated at set intervals to confirm that the oxygen partial pressure of the harvested liquid is within the process range.

[0079] Before sampling begins, the three-way valve 82 is first connected to the venting line 83 to push all the residual liquid in the L-shaped line 81 back into the tank. Then, the three-way valve 82 disconnects the venting line 83 and connects the sampling line 84 for sampling. This operation not only reduces waste discharge but also ensures the authenticity and representativeness of the samples. When the oxygen partial pressure measured by the sampling results is low, the sampling line 84 is disconnected and the venting line 83 is connected. The gas flow controller (MFC) 85 installed on the venting line 83, based on the oxygen partial pressure detection value, introduces a specific type and flow rate of gas into the harvest liquid in the harvest tank 6 through the L-shaped line 81 until the oxygen partial pressure value rises to within the process requirements range.

[0080] In this embodiment, an oxygen content sensor 86 is provided in the harvest tank 6. The oxygen content sensor 86 is configured to be electrically connected to the gas flow controller (MFC) 85. The gas flow controller (MFC) 85 automatically adjusts the amount of oxygen or air input in the ventilation pipeline 83 according to the oxygen partial pressure of the harvest liquid detected by the oxygen content sensor 86 in the harvest tank.

[0081] In this embodiment, the oxygen content sensor 86 is a dissolved oxygen sensor.

[0082] The ventilation gas described in this embodiment can be selected from one or more of oxygen, nitrogen, carbon dioxide, or air.

[0083] The harvesting system for clarifying cell culture medium provided by this utility model has been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of this utility model. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of this utility model. It should be noted that those skilled in the art can make several improvements and modifications to this utility model without departing from the principles of this utility model, and these improvements and modifications also fall within the protection scope of the claims of this utility model.

Claims

1. A cell culture medium clarification and harvesting system, mainly comprising a bioreactor, a power pump, a primary membrane, a secondary membrane, a feed sterilizing filter, a harvesting tank, and a discharge sterilizing filter, characterized in that, The clarification and harvesting system has a pressure regulating device between the secondary membrane and the feed sterilization filter to regulate the pressure at the inlet of the primary membrane.

2. The clarification and harvesting system according to claim 1, characterized in that, The pressure regulating device is a buffer system, which regulates the pressure at the primary membrane inlet by controlling the amount of fluid flowing through the buffer system.

3. The clarification and harvesting system according to claim 2, characterized in that, The buffer system is a buffer bag, which is configured to buffer the fluid delivered through the secondary membrane outlet, and the amount of fluid in the buffer bag is adjusted by its own volume change or a peristaltic pump.

4. The clarification and harvesting system according to claim 3, characterized in that, The buffer bag is a flexible bag that can automatically adjust its internal volume according to the internal pressure or fluid volume.

5. The clarification and harvesting system according to claim 4, characterized in that, The buffer bag is equipped with a feed pipe, the proximal end of which is connected to the outlet of the secondary membrane, and the distal end of which extends into the buffer bag.

6. The clarification and harvesting system according to claim 5, characterized in that, The buffer system is further provided with a peristaltic pump at the outlet of the buffer bag, the peristaltic pump being configured to pump fluid from the buffer bag into the feed sterilization filter.

7. The clarification and harvesting system according to claim 6, characterized in that, The buffer bag is also equipped with a weighing sensor, which transmits the weight signal of the fluid in the buffer bag to the PID. The PID automatically sets the pump speed of the peristaltic pump according to the fluid weight and automatically adjusts the amount of fluid entering the feed sterilization filter.

8. The clarification and harvesting system according to claim 2, characterized in that, The buffer system is a diaphragm pump, which is configured to automatically adjust its pumping speed according to the pressure at the outlet of the secondary membrane.

9. The clarification and harvesting system according to any one of claims 1-8, characterized in that, A ventilation system is provided at the bottom of the harvesting tank, consisting of an L-shaped pipe, a three-way valve, a ventilation pipe, and a sampling pipe installed on the harvesting tank. The distal end of the L-shaped pipe extends into the harvesting tank, and the proximal end is connected to the ventilation pipe and the sampling pipe respectively through the three-way valve. The angle formed by the distal end and the proximal end of the L-shaped pipe is >90°. The three-way valve controls the connection and disconnection between the ventilation pipeline and the sampling pipeline and the harvest tank; A gas flow controller (MFC) may also be installed on the ventilation pipeline. The gas flow controller (MFC) is configured to adjust the oxygen content of the gas supplied to the harvest tank in real time based on the oxygen partial pressure of the harvest liquid in the harvest tank.

10. The clarification and harvesting system according to claim 9, characterized in that, An oxygen content sensor is installed inside the harvesting tank. The oxygen content sensor is configured to be electrically connected to the gas flow controller (MFC) to detect the oxygen partial pressure of the harvested liquid in the harvesting tank in real time and transmit the oxygen partial pressure to the gas flow controller (MFC), so that the gas flow controller (MFC) can automatically regulate the amount of oxygen input in the ventilation pipeline.

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