Alternating tangential flow perfusion system and control method therefor

Abstract

The present invention relates to an alternating tangential flow perfusion system and a control method therefor. The alternating tangential flow perfusion system comprises: a consumable assembly, a weighing assembly, an inflation assembly, a gas extraction assembly, and a control box, wherein the consumable assembly is disposed on the weighing assembly, the weighing assembly is used for monitoring the weight of a fluid in the consumable assembly in real time, and the control box determines the motion state of an elastic balloon on the basis of the variation of the weight of the fluid and the rate of change of the weight of the fluid, so as to control the operating states of the inflation assembly and the gas extraction assembly. The alternating tangential flow perfusion system and the control method therefor are provided in order to overcome existing defects, such that the operation of the system is more stable and reliable.

Description

Alternating tangential flow perfusion system and control method thereof TECHNICAL FIELD

[0001] The present application relates to an alternating tangential flow perfusion system and a control method thereof. BACKGROUND

[0002] In recent years, the biopharmaceutical industry often uses an alternating tangential flow perfusion system to obtain high-density cell culture results. The alternating tangential flow perfusion system uses an alternating method to push the culture solution through the filter membrane, has the advantages of good membrane filament flushing effect, low shear force, and high working efficiency. The alternating tangential flow perfusion system is widely used in the biopharmaceutical industry, especially in the fields of monoclonal antibody production, virus particle production, and biological drug harvesting. Generally, the alternating tangential flow perfusion system generates varying power by driving the membrane with vacuum and compressed air, and the membrane pushes the culture medium to flow back and forth to flush the hollow fiber. During the driving process, the pressure sensor detects the pressure change of the driving membrane to determine whether the membrane has reached the limit position. Once the pressure sensor detects a sudden change in pressure, it is considered that the membrane has reached the limit position, and the control system performs a switching action to switch the flushing direction.

[0003] However, it is found in actual use that the existing alternating tangential flow perfusion system is not stable and often malfunctions. SUMMARY

[0004] The purpose of the present application is to overcome the existing defects and provide an alternating tangential flow perfusion system and a control method thereof, so that the system runs more stably and reliably.

[0005] The technical solution to achieve the above-mentioned purpose is: an alternating tangential flow perfusion system, comprising: a consumable assembly, a weighing assembly, an inflation assembly, a gas extraction assembly, and a control box;

[0006] The consumable assembly includes a rigid transparent spherical shell, an elastic balloon, and a hollow fiber column. The elastic balloon is arranged in the rigid transparent spherical shell, the inner cavity of the elastic balloon is in communication with the hollow fiber column connected to the rigid transparent spherical shell, the butt joint of the hollow fiber column is connected to the reactor through a first infusion pipeline, and the upper end of the hollow fiber column is connected to a waste liquid bottle through a second infusion pipeline.

[0007] The consumable assembly is arranged on the weighing assembly, and the weighing assembly is used to monitor the weight of the fluid in the consumable assembly in real time.

[0008] The weighing assembly, the inflation assembly and the air extraction assembly are connected with the control box, the inflation assembly is used for inflating between the elastic balloon and the rigid transparent spherical shell, so that the elastic balloon shrinks under the action of pressure difference, the air extraction assembly is used for extracting air from between the elastic balloon and the rigid transparent spherical shell, so that the elastic balloon expands under the action of pressure difference, the control box judges the motion state of the elastic balloon according to the change amount of fluid weight and the change rate of fluid weight, and then controls the working state of the inflation assembly and the air extraction assembly.

[0009] Optionally, the control box comprises a gas path control module, the gas path control module has an inflation path and an air extraction path, the inflation path is sequentially provided with an inflation port, a precision pressure regulating valve, a first pressure proportional valve, a gas flow meter and a gas control electromagnetic valve, and the air extraction path is sequentially provided with an air extraction port, a vacuum control valve, a second pressure proportional valve, a gas flow meter and a gas control electromagnetic valve.

[0010] Optionally, the inflation assembly comprises a compressed air source and a compressed air connecting pipeline, one end of the compressed air connecting pipeline is connected with the compressed air source, the other end of the compressed air connecting pipeline is connected with the inflation port of the control box, and the gas path output end of the control box is connected with the consumable assembly through a third connecting pipeline.

[0011] Optionally, the air extraction assembly comprises a vacuum pump and a second connecting pipeline, one end of the second connecting pipeline is connected with the vacuum pump, and the other end of the second connecting pipeline is connected with the air extraction port of the control box.

[0012] Optionally, the consumable assembly is further connected with a support assembly, the lower end of the support assembly is fixed on the weighing assembly, and the support assembly is used for supporting the consumable assembly.

[0013] Optionally, a first peristaltic pump is connected on the second infusion pipeline, and the reactor is communicated with a liquid supplementing bottle through a third infusion pipeline, and a second peristaltic pump is connected on the third infusion pipeline.

[0014] Optionally, the upper end of the rigid transparent spherical shell is communicated with the hollow fiber column, and the upper end of the hollow fiber column is provided with the docking port.

[0015] Optionally, the consumable assembly further comprises an air inlet flange, a screen and an elbow;

[0016] The upper end of the shell of the hollow fiber column is provided with a first port, the lower end is provided with a second port, the upper end of the side wall of the shell is provided with a third port, and the lower end of the side wall is provided with a fourth port.

[0017] The rigid transparent spherical shell is connected to the third interface through a first chuck, and the other end of the rigid transparent spherical shell is connected to the air inlet flange; the elastic balloon is sleeved on the third interface through an opening of the elastic balloon; the screen is arranged in the inner part of the shell, and one end of the elbow is connected to the second interface;

[0018] When the elastic balloon expands under the action of pressure difference, fluid flows from the elbow to the shell and flows into the elastic balloon through the screen;

[0019] When the elastic balloon shrinks under the action of pressure difference, fluid flows from the elastic balloon to the shell and flows back to the elbow through the screen.

[0020] Optionally, the rigid transparent spherical shell is connected to the air inlet flange through a second chuck, one end of the air inlet flange is a chuck interface for matching the second chuck, and the other end is an internally threaded opening for being connected to a connecting pipeline joint.

[0021] Optionally, a support is further arranged in the elastic balloon, a threaded hole is arranged on the third interface, one end of the support is connected to the threaded hole, and the other end of the support is a flange edge for fixing the position of the elastic balloon.

[0022] Optionally, a fine hole is arranged on the third interface for connecting the elastic balloon, and the fine hole surrounds the threaded hole.

[0023] Optionally, the elastic balloon is a spherical silica gel material membrane, and the screen is a stainless steel screen.

[0024] Optionally, both ends of the elbow, the first interface and the second interface are chuck interfaces.

[0025] Optionally, the screen has a tubular structure with an open end and a closed end, and the open end of the screen faces the elbow.

[0026] Optionally, a stepped surface is arranged on the open end of the screen and the second interface, and the stepped surface of the screen matches the stepped surface of the second interface.

[0027] Optionally, the closed end of the screen is higher than the third interface, and the mesh size of the screen is smaller than the size of the microcarrier.

[0028] Optionally, the method further comprises a connecting pipeline, a third infusion pipeline, a fourth infusion pipeline and a first peristaltic pump.

[0029] One end of the second infusion pipeline is connected to the first interface at the upper end of the shell, and the first peristaltic pump is connected to the second infusion pipeline.

[0030] One end of the connecting pipeline is connected with the air inlet flange, and the other end of the connecting pipeline is connected with the control box.

[0031] One end of the fourth infusion pipeline is connected with the fourth interface of the lower end side wall of the shell.

[0032] One end of the first infusion pipeline is connected with the elbow, and the other end is connected with the reactor, and the reactor is connected with a third infusion pipeline.

[0033] Correspondingly, the application also provides a control method of the alternating tangential flow perfusion system, which comprises the following steps:

[0034] Step S1, inflating the rigid transparent spherical shell by the inflating assembly to completely shrink the elastic balloon;

[0035] Step S2, deflating the rigid transparent spherical shell by the deflating assembly to completely inflate the elastic balloon;

[0036] Step S3, continuously operating step S1 and step S2 until the cell perfusion culture process is completed.

[0037] The step S1 comprises:

[0038] Step S11, in the initial state, the elastic balloon is completely inflated, the control box controls the compressed air connecting pipeline to inflate the rigid transparent spherical shell, so that the elastic balloon gradually shrinks; in this process, the weighing assembly is used to monitor the fluid weight in the consumable assembly in real time;

[0039] When the change amount of the fluid weight is greater than or equal to the first set value, the change rate of the fluid weight is compared with the second set value, and if the change rate of the fluid weight is less than or equal to the second set value, it is considered that the elastic balloon has been completely deflated, and the switching condition is reached;

[0040] Step S12, closing the compressed air connecting pipeline and recording the actual action time of the inflation operation;

[0041] Before the next inflation operation, the actual action time of the inflation operation is compared with the theoretical action time;

[0042] If the actual action time is greater than the theoretical action time, the control box increases the driving force of the next inflation operation through the air path control module; if the actual action time is less than the theoretical action time, the control box reduces the driving force of the next inflation operation through the air path control module.

[0043] The step S2 comprises:

[0044] Step S21, the control box controls the vacuum pump to pump air out of the rigid transparent spherical shell, so that the elastic balloon gradually expands; in this process, the weight of the fluid in the consumable assembly is monitored in real time by the weighing assembly;

[0045] When the change amount of the fluid weight is greater than or equal to the third set value, the change rate of the fluid weight is compared with the fourth set value, and if the change rate of the fluid weight is less than or equal to the fourth set value, it is considered that the elastic balloon has been completely inflated, and the switching condition is reached;

[0046] Step S22, the vacuum pump is turned off, and the actual operation time of the air pumping operation is recorded.

[0047] Before the next air pumping operation, the actual operation time of the air pumping operation is compared with the theoretical operation time;

[0048] If the actual operation time is greater than the theoretical operation time, the control box increases the driving force of the next air pumping operation through the air path control module; if the actual operation time is less than the theoretical operation time, the control box reduces the driving force of the next air pumping operation through the air path control module.

[0049] During the cyclic switching of the inflation operation and the air pumping operation, if the actual operation time exceeds the preset time and the switching condition is not reached, the forced switching is performed according to the operation timeout logic.

[0050] The beneficial effects of the present application are: the alternating tangential flow irrigation system and the control method thereof can solve the problem that when the existing alternating tangential flow irrigation system operates, the pressure sensor detects a sudden change in pressure, and the control system performs a switching action, because air can be compressed, the input pressure of the gas will change with the input flow, and the pressure will not suddenly change after reaching the theoretical switching point, or the pressure will suddenly change before reaching the theoretical switching point, resulting in that the time for the control system to drive the balloon to move is not the same, and the system is unstable. At the same time, since the switching logic uses the weight derivative as the main switching condition, the problem that the weight drifts due to uncontrollable conditions during the use of the device, thereby affecting the operation of the device, can be effectively solved.

[0051] In addition, the device uses the combination of weight change and weight change rate as the switching condition, and does not use weight as a separate switching condition, which can effectively avoid the situation that the elastic balloon or the diaphragm of the diaphragm pump does not move to the limit position, and the weight cannot reach the peak or valley, thereby affecting the continuous operation of the device. BRIEF DESCRIPTION OF DRAWINGS

[0052] Fig. 1 is a flowchart of the weight switching logic method of the alternating tangential flow irrigation system of the present application;

[0053] Fig. 2 is a structural schematic diagram of the alternating tangential flow irrigation system of an embodiment of the present application;

[0054] Figure 3 is a weight and pressure curve of the alternating tangential flow perfusion system of the present application in operation;

[0055] Figure 4 is a schematic diagram of the movement of the elastic balloon of the alternating tangential flow perfusion system of the present application;

[0056] Figure 5 is a schematic diagram of the control method of the alternating tangential flow perfusion system of the present application;

[0057] Figure 6 is a schematic diagram of the gas path control module of the alternating tangential flow perfusion system of the present application;

[0058] Figure 7 is a schematic diagram of the hollow fiber column of the consumable assembly of another embodiment of the present application;

[0059] Figure 8 is a sectional view of the consumable assembly of another embodiment of the present application;

[0060] Figure 9 is a schematic diagram of the alternating tangential flow perfusion system of another embodiment of the present application.

[0061] Figure: 1, consumable assembly; 2, support assembly; 3, weighing assembly; 4, control box; 5, vacuum pump; 6, compressed air connection pipeline; 7, second connection pipeline; 8, third connection pipeline; 9, first peristaltic pump; 10, reactor; 11, first infusion pipeline; 12, second infusion pipeline; 13, third infusion pipeline; 14, waste liquid bottle; 15, liquid supplement bottle; 16, second peristaltic pump; 18, fourth infusion pipeline; 101, rigid transparent spherical shell; 102, air inlet flange; 103, strut; 104, elastic balloon; 105, hollow fiber column; 1051, outer shell; 106, screen; 107, elbow; 108, first interface; 109, second interface; 110, third interface; 111, fourth interface; 112, first chuck; 113, second chuck; 114, threaded hole; 115, fine hole; 601, air extraction port; 602, vacuum control valve; 603, second pressure proportional valve; 604, pneumatic electromagnetic valve; 605, gas path output port; 606, gas flow meter; 607, first pressure proportional valve; 608, precision pressure regulating valve; 609, air charging port. DETAILED DESCRIPTION

[0062] The technical solutions of the present application will be clearly and completely described below with reference to the drawings. In the description of the present application, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer" and the like indicate the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and therefore cannot be understood as indicating or implying that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the present application. In addition, the terms "first", "second", "third" are only for descriptive purposes and cannot be understood as indicating or implying opposite importance.

[0063] The present application will be further described below with reference to the drawings.

[0064] As shown in FIGS. 1-4, the alternating tangential flow irrigation system of an embodiment of the present application comprises a consumable assembly 1, a weighing assembly 3, an air charging assembly, an air pumping assembly, and a control box 4; the consumable assembly 1 comprises a rigid transparent spherical shell 101, an elastic balloon 104, and a hollow fiber column 105, the elastic balloon 104 is arranged in the rigid transparent spherical shell 101, the inner cavity of the elastic balloon 104 is in communication with the hollow fiber column 105 connected to the upper end of the rigid transparent spherical shell 101, the upper end of the hollow fiber column 105 is connected to the reactor 10 through the first liquid conveying pipeline 11, and the upper end of the hollow fiber column 105 is connected to the waste liquid bottle 14 through the second liquid conveying pipeline 12; the consumable assembly 1 is connected to the weighing assembly 3, and the weighing assembly 3 is used to monitor the weight of the fluid in the consumable assembly 1 in real time.

[0065] Specifically, the weighing assembly 3, the air charging assembly, and the air pumping assembly are all connected to the control box 4, the air charging assembly is used to charge air between the elastic balloon 104 and the rigid transparent spherical shell, so that the elastic balloon 104 shrinks under the action of pressure difference, the air pumping assembly is used to pump air from between the elastic balloon 104 and the rigid transparent spherical shell 101, so that the elastic balloon 104 expands under the action of pressure difference, the control box 4 judges the motion state of the elastic balloon 104 according to the change of the fluid weight monitored by the weighing assembly 3 and the change rate of the fluid weight, and then controls the working state of the air charging assembly and the air pumping assembly.

[0066] As shown in FIG. 6, the control box 4 comprises an air path control module, the air path control module has an air charging path and an air pumping path, the air charging path is sequentially provided with an air charging port 609 (compressed air connection port), a precision pressure regulating valve 608, a first pressure proportional valve 607, a gas flow meter 606, and a gas control electromagnetic valve 604, and the air pumping path is sequentially provided with an air pumping port 601 (vacuum air source connection port), a vacuum control valve 602, a second pressure proportional valve 603, a gas flow meter 606, and a gas control electromagnetic valve 604.

[0067] Specifically, the inflation assembly includes a compressed air source and a compressed air connecting pipeline 6, one end of the compressed air connecting pipeline 6 is connected with the compressed air source, the other end of the compressed air connecting pipeline 6 is connected with the inflation port 609 of the control box 4, and the gas path output end of the control box 4 is connected with the consumable assembly 1 through a third connecting pipeline 8.

[0068] Specifically, the space between the elastic balloon 104 and the rigid transparent spherical shell is communicated with the gas path output end of the control box 4 through the third connecting pipeline 8. The rigid transparent spherical shell 101 or the connecting pipeline below the rigid transparent spherical shell 101 can be provided with a vent hole, and the third connecting pipeline 8 is sealingly connected with the vent hole.

[0069] Specifically, the exhaust assembly includes a vacuum pump 5 and a second connecting pipeline 7, one end of the second connecting pipeline 7 is connected with the vacuum pump 5, and the other end of the second connecting pipeline 7 is connected with the exhaust port of the control box 4.

[0070] Specifically, the consumable assembly 1 is further connected with a support assembly 2, the lower end of the support assembly 2 is fixed on the weighing assembly 3, and the support assembly 2 is used for supporting the consumable assembly 1.

[0071] Specifically, the second liquid conveying pipeline 12 is connected with a first peristaltic pump 9, the reactor 10 is communicated with a liquid supplement bottle 15 through a third liquid conveying pipeline 13, and the third liquid conveying pipeline 13 is connected with a second peristaltic pump 16. The control of the first peristaltic pump 9 and the second peristaltic pump 16 can be separately set, can be integrated with the electric control unit of the reactor, or can be integrated in the control box 4. The first peristaltic pump 9 is used for extracting waste liquid, and the second peristaltic pump 16 is used for supplementing new liquid. Generally, the extraction amount of the waste liquid and the supplement amount of the new liquid remain consistent, so as to ensure that the liquid quality in the reactor 10 remains unchanged.

[0072] As shown in FIG. 2, the elastic balloon 104 inside the consumable assembly 1 is periodically driven through the control box 4, so as to control the periodic exchange of the liquid (i.e. the culture medium) between the inside of the hollow fiber column 105 and the inside of the reactor 10. At the same time, the first peristaltic pump 9 is used to continuously extract the used culture medium which has been intercepted by the hollow fiber column 105 from the inside of the hollow fiber column 105, and the second peristaltic pump 16 is used to pump the fresh culture medium in the liquid supplement bottle into the reactor 10, so as to achieve the purpose of the exchange of the culture medium in the reactor.

[0073] During the operation, the control box 4 calculates the weight change rate according to the real-time monitoring of the liquid weight by the weighing assembly 3, respectively obtains the change curves of the weight and the weight derivative, and judges whether the expansion or contraction of the elastic balloon reaches the limit position according to the preset change amount parameter and the change rate parameter, and then decides whether to perform the switching operation.

[0074] As shown in FIG. 3, in one action cycle, when the ball in the consumable assembly 1 is completely squeezed, at this time the liquid weight in the consumable assembly 1 and its pipeline is the lowest, denoted as W1, the suction action is performed, the elastic balloon 104 starts to expand under the action of pressure difference, the liquid flows into the elastic balloon 104, and the liquid weight in the consumable assembly 1 and its pipeline gradually increases, when the change amount of the weight is greater than or equal to the first set value (i.e., real-time weight-W1≥△W), the change rate of the liquid weight is compared with the second set value, if the change rate of the weight is less than or equal to the second set value, it is considered that the ball has been completely inflated, at this time the liquid weight in the consumable assembly 1 and its pipeline is the maximum, denoted as W2, then the inflation action is switched, the elastic balloon 104 starts to shrink under the action of pressure difference, the liquid flows out of the elastic balloon 104, and the liquid weight in the consumable assembly 1 and its pipeline gradually decreases, when the change amount of the liquid weight is greater than or equal to the third set value (i.e., W2-real-time weight≥△W), the change rate of the liquid weight is compared with the fourth set value, if the change rate of the liquid weight is less than or equal to the fourth set value, it is considered that the ball has been completely squeezed, and the suction action is switched again.

[0075] Among them, the first set value and the third set value are both change amount parameters of the liquid weight, and the second set value and the fourth set value are both change rate parameters of the liquid weight.

[0076] In this embodiment, the first set value and the third set value are equal, both of which are △W. △W can be set according to the maximum weight W2 of the fluid in the consumable assembly 1, and is usually set to 60%-80% of the maximum weight W2. For example, the maximum weight W2 is 100 grams, and △W can be set to 80% of the maximum weight, i.e. 80 grams. In this way, when the fluid weight decreases from 100 grams to 20 grams, the change rate of the fluid weight starts to be calculated.

[0077] In this embodiment, the second set value and the fourth set value are equal, both of which are △V. △V can be set according to the measurement error of the fluid weight change rate, which comes from the measurement error of the weighing assembly, or is caused by the shaking of the consumable assembly 1 when the elastic balloon is moving at a higher speed. For example, △V is set to ±0.1 gram / second, when the change rate of the fluid weight decreases to 0.1 gram / second, it can be considered that the fluid weight is no longer changing, reaching the switching condition.

[0078] In addition, the control box 4 will also compare the action time with the theoretical time to determine whether the elastic balloon 104 is moving too fast or too slow, so as to adjust the driving force of the equipment, and reduce or increase the driving force, so as to control the liquid movement speed inside the hollow fiber. In each action process, if the switching condition is not reached for a long time, the forced switching will be performed according to the action timeout logic.

[0079] The alternating tangential flow perfusion system provided by the embodiment can solve the problem that the existing alternating tangential flow perfusion system cannot perform switching action after the pressure sensor detects a sudden change in pressure and the control system is executed, because air can be compressed, the input pressure of the gas will change with the input flow, and the sudden change in pressure will not occur after reaching the theoretical switching point, or the sudden change in pressure will occur in advance, resulting in that the time for the control system to drive the diaphragm to move is not the same, and the system is unstable. At the same time, because the switching logic used takes the weight derivative as the main switching condition, the problem that the weight drifts due to uncontrollable conditions during use of the device, thereby affecting the operation of the device, can be effectively solved.

[0080] In addition, the device takes the weight change and the weight change rate as the switching conditions, and not the weight as a separate switching condition, which can effectively avoid the situation that the elastic balloon does not move to the limit position, the weight cannot reach the peak or valley, and the continuous action of the device is affected.

[0081] Correspondingly, the embodiment also provides a control method of the alternating tangential flow perfusion system. As shown in FIG. 5, the control method of the alternating tangential flow perfusion system comprises the following steps:

[0082] Step S1, inflating the rigid transparent spherical shell 101 by the inflating assembly to make the elastic balloon 104 completely shrink;

[0083] Step S2, deflating the rigid transparent spherical shell 101 by the deflating assembly to make the elastic balloon 104 completely expand;

[0084] Step S3, continuously operating step S1-Step S2 until the cell perfusion culture process is completed.

[0085] Specifically, step S1 comprises:

[0086] Step S11, in the initial state, the elastic balloon 104 is in a completely inflated state, the control box 4 controls the compressed air connection pipeline 6 to inflate the rigid transparent spherical shell 101, so that the elastic balloon 104 gradually shrinks; in this process, the weighing assembly 3 monitors the weight of the fluid in the consumable assembly 1 in real time;

[0087] When the change amount of the fluid weight is greater than or equal to the first set value, the change rate of the fluid weight is calculated, and if the change rate of the fluid weight is less than or equal to the second set value, it is considered that the elastic balloon 104 has been completely deflated, and the switching condition is reached;

[0088] Step S12, closing the compressed air connection pipeline 6, and recording the actual action time of the inflation operation.

[0089] After the inflation operation is completed, before the next inflation operation, it also includes: comparing the actual operation time of the inflation operation with the theoretical operation time;

[0090] If the actual operation time is greater than the theoretical operation time, the control box 4 increases the driving force of the next inflation operation through the air path control module; if the actual operation time is less than the theoretical operation time, the control box 4 reduces the driving force of the next inflation operation through the air path control module.

[0091] Step S2 includes:

[0092] Step S21, the control box 4 controls the vacuum pump 5 to pump out the rigid transparent spherical shell, so that the elastic balloon 104 gradually expands; in this process, the fluid weight in the consumable assembly 1 is monitored in real time through the weighing assembly 3;

[0093] When the change amount of the fluid weight is greater than or equal to the third set value, the change rate of the fluid weight is calculated, and if the change rate of the fluid weight is less than or equal to the fourth set value, it is considered that the elastic balloon 104 has been completely inflated, and the switching condition is reached;

[0094] Step S22, the vacuum pump 5 is closed, and the actual operation time of the pumping operation is recorded.

[0095] After the pumping operation is completed, before the next pumping operation, it also includes: comparing the actual operation time of the pumping operation with the theoretical operation time;

[0096] If the actual operation time is greater than the theoretical operation time, the control box 4 increases the driving force of the next pumping operation through the air path control module; if the actual operation time is less than the theoretical operation time, the control box 4 reduces the driving force of the next pumping operation through the air path control module.

[0097] The theoretical operation time is set according to the time requirement of the cell perfusion culture process. If the cell perfusion culture process requires a time period of t1 for each inflation operation, the theoretical operation time of the inflation operation is t1. Similarly, if the cell perfusion culture process requires a time period of t2 for each pumping operation, the theoretical operation time of the pumping operation is t2. The theoretical operation time t2 of each pumping operation and the theoretical operation time t1 of the inflation operation can be equal.

[0098] During the cyclic switching process of the inflation operation and the pumping operation, if the actual operation time exceeds the preset time and the switching condition is not reached, forced switching is performed according to the operation timeout logic.

[0099] If the theoretical action time of each inflation operation and each exhaust operation is t, the preset time can be set between 1.1t and 1.3t. For example, the preset time is 1.2t, when the actual action time of the inflation operation or the exhaust operation reaches 1.2t, the switching condition is still not met, and the driving force is increased to directly force the switching.

[0100] The alternating tangential flow perfusion system and the control method thereof provided by the application detect how much liquid enters the pipeline (including the hollow fiber column 105 and the elastic balloon 104) of the consumable assembly 1 by placing a weighing assembly 3 at the bottom of the consumable assembly 1 to weigh the consumable assembly 1 in real time. The weight of the liquid in the pipeline of the consumable assembly 1 changes in real time during the movement of the elastic balloon 104. When the elastic balloon 104 moves to the limit position, the liquid in the pipeline of the consumable assembly 1 basically no longer flows, the flow rate approaches zero, and the weight of the consumable assembly 1 no longer changes.

[0101] In the consumable assembly 1, the internal volume of the hollow fiber column is fixed, and the volume of the elastic balloon 104 can change between the maximum volume and the minimum volume, so the volume change of the elastic balloon 104 can be judged by the weight change amplitude (for example: during the movement of the elastic balloon 104, the hollow fiber column always maintains a full state, if the volume of the elastic balloon 104 when fully inflated is X, and when fully deflated, the volume of the elastic balloon 104 changes to 0, at this time, X volume enters the hollow fiber column 105, and at the same time, due to the fixed volume of the hollow fiber column 105 and the full state before X enters, the X volume of liquid entering will cause the X volume of liquid originally in the hollow fiber column 105 to be expelled, and since the hollow fiber column 105 is connected to the reactor 10, the expelled liquid enters the reactor 10. During the entire operation, the total volume of the liquid inside the hollow fiber column 105 does not change, and the volume of the liquid inside the elastic balloon 104 changes. Therefore, when the weight change amplitude of the consumable assembly 1 does not reach the set value, it means that the elastic balloon 104 is not fully inflated or fully deflated.

[0102] The inventor found that the existing alternating tangential flow irrigation system detects the pressure change of the membrane by a pressure sensor and then performs switching action. The premise of this control method is that the membrane reaches the limit position (contact with the upper or lower spherical shell) and the membrane position is not moving, and the compressed air or vacuum is still continuously outputting power to the membrane to form a pressure change. However, the pressure change only reflects that the membrane reaches the limit position, and has nothing to do with whether there is liquid in the ball and whether the volume of the liquid changes. If the control system drives the membrane to move to suck in the liquid and an abnormal situation occurs, there is actually no liquid in the ball, and the pressure change will still occur when the membrane reaches the limit position. Thus, the control system will make a wrong judgment and think that the switching condition has been reached to reverse the liquid output, resulting in a misoperation. Therefore, this control method can only ensure that the driving power of the membrane is relatively regular, and cannot determine whether the liquid suction and discharge is consistent with the movement of the membrane. Only the change of air pressure is measured, and the result is not reliable. Moreover, since air can be compressed, the input pressure of the gas will change with the change of the input flow, so the pressure change may not occur at the theoretical switching point or may occur in advance. Thus, the control system drives the membrane to move for different lengths of time, and the system runs unstably.

[0103] To solve the above problems, the present application monitors the weight change of the consumable assembly (actually the weight change of the liquid inside) in real time, and determines whether the elastic balloon of the consumable assembly reaches the limit position according to whether the weight reaches the peak value and the valley value.

[0104] It is considered that the measurement deviation and drift of the weighing assembly will cause misjudgment. For example, when the elastic balloon performs a contraction action, the measurement deviation and drift of the weighing assembly will cause the measured value to fail to reach the predetermined valley value or to reach the predetermined valley value too early, thereby affecting the normal operation of the device. When the elastic balloon performs an expansion action, the measurement deviation or drift of the weighing assembly will cause the measured value to fail to reach the predetermined peak value or to reach the predetermined peak value too early, thereby affecting the normal operation of the device. In addition, the liquid in the elastic balloon will have a certain inertial impact when it reaches the limit position, although the total amount of the liquid does not change, but the inertial movement will cause the weighing assembly to measure a larger peak weight and a smaller valley weight, so that the liquid is not completely discharged or not completely filled, and the system judges that the switching condition has been reached. Therefore, the present application not only uses the weight change as a judgment index, but also uses the weight change rate as another judgment index, and combines the two indexes to determine whether the movement of the elastic balloon reaches the limit state.

[0105] Specifically, when the elastic balloon is completely deflated or completely inflated, the rate of change of the weight of the liquid will become smaller, and when the rate of change of the weight approaches zero, it represents that the amount of liquid in the elastic balloon no longer changes. By combining the change amount of the weight and the rate of change of the weight as the switching condition, the measurement deviation problem caused by the weighing drift and the liquid inertia impact can be effectively avoided, thereby ensuring that the liquid inside the elastic balloon can be completely discharged or filled when the switching action is performed, so as to ensure that the alternating tangential flow control system is more stable and reliable.

[0106] In the alternating tangential flow perfusion system, the alternating tangential flow filtration (ATF) technology is used to periodically change the flow direction to reduce cell accumulation and pollution, thereby improving the filtration efficiency and cell activity. Compared with the traditional tangential flow filtration (TFF), the ATF system shows better performance in cell activity and product retention, which can prolong the process duration and reduce cell accumulation and product retention. The existing alternating tangential flow perfusion system is mainly applied to suspended cells, and currently there is a lack of a simple and effective practical structure for adherent cell perfusion and microcarrier perfusion, and there is a lack of a reliable solution for microcarrier sedimentation and gas aggregation.

[0107] FIGS. 7-9 are structural schematic diagrams of an alternating tangential flow perfusion system and a consumable assembly of another embodiment of the present application. The main difference between this another embodiment and the embodiment shown in FIG. 2 is that the structure of the consumable assembly is different, which is simple in structure and convenient to install, and can effectively combine the alternating tangential flow perfusion process with adherent cell culture.

[0108] As shown in FIGS. 7-8, the consumable assembly forms an alternating tangential flow filtration structure, which includes a rigid transparent spherical shell 101, a gas inlet flange 102, an elastic balloon 104, a hollow fiber column 105 with an outer shell 1051, a screen 106, and an elbow 107; the upper end of the outer shell 1051 is provided with a first interface 108 (used as an upper end liquid outlet connected with a waste liquid bottle), the lower end is provided with a second interface 109, the upper end of the sidewall of the outer shell 1051 is provided with a third interface 110, and the lower end of the sidewall is provided with a fourth interface 111; the rigid transparent spherical shell 101 is connected to the third interface 110 through a first chuck 112, and the other end of the rigid transparent spherical shell 101 is connected to the gas inlet flange 102; the elastic balloon 104 is arranged inside the rigid transparent spherical shell 101 and is sleeved on the third interface 110 through an opening thereof; the screen 106 is arranged inside the outer shell 1051, one end of the elbow 107 (used as a connecting interface connected with the reactor 10) is connected with the second interface 109; and the inner space of the elastic balloon 104 is in communication with the inner space of the outer shell 1051 through the third interface 110.

[0109] Specifically, when the elastic balloon 104 expands under the action of pressure difference, the fluid flows from the elbow 107 to the shell 1051, and flows into the elastic balloon 104 through the screen 106; the screen 106 is used to intercept the microcarriers in the fluid, so as to avoid the microcarriers entering the elastic balloon 104.

[0110] Specifically, when the elastic balloon 104 contracts under the action of pressure difference, the fluid flows from the elastic balloon 104 to the shell 1051, and flows back to the elbow 107 through the screen 106.

[0111] Specifically, the rigid transparent spherical shell 101 is connected to the gas inlet flange 102 through the second chuck 113. One end of the gas inlet flange 102 is a chuck interface for matching the second chuck 113, and the other end is a female threaded port for connecting to the joint of the connecting pipeline 8.

[0112] Specifically, it also includes a support 103, the support 103 is arranged in the elastic balloon 104, a threaded hole 114 is arranged on the third interface 110, one end of the support 103 is connected with the threaded hole 114, and the other end is a flange edge for fixing the position of the elastic balloon 104. A fine hole 115 is arranged on the third interface 110 for communicating the elastic balloon 104, and the fine hole 115 surrounds the threaded hole 114. The elastic balloon 104 is a spherical silica gel material diaphragm, and the screen 106 is a stainless steel screen. Both ends of the elbow 107, the first interface 108 and the second interface 109 are chuck interfaces. The screen 106 has a cylindrical structure with one open end and the other closed end, and the open end of the screen 106 faces the elbow 107.

[0113] Specifically, the open end of the screen 106 and the second interface 109 are both provided with a stepped surface, and the stepped surface of the screen 106 cooperates with the stepped surface of the second interface 109 for alignment and installation. The closed end of the screen 106 is higher than the third interface 110, and the mesh size of the screen 106 is smaller than the size of the microcarriers. The microcarriers refer to beads with a diameter of 60-250 μm, which can be suitable for the growth of adherent cells. Generally composed of natural dextran or various synthetic polymers.

[0114] As shown in Figure 9, the perfusion culture system based on the alternating tangential flow filtration structure includes: a second infusion pipeline 12 (as an upper filtrate pipeline), a connecting pipeline 8, a fourth infusion pipeline 18 (as a lower filtrate pipeline), a first infusion pipeline 11, a first peristaltic pump 9, a control box 4, a reactor 10 and the control box 4; one end of the second infusion pipeline 12 is connected to a first interface 108 at the upper end of a shell 1051, and the first peristaltic pump 9 is connected to the second infusion pipeline 12; one end of the connecting pipeline 8 is connected to an air inlet flange 102, and the other end of the connecting pipeline 8 is connected to the control box 4; one end of the fourth infusion pipeline 18 is connected to a fourth interface 111 at the lower end of the sidewall of the shell 1051; one end of the first infusion pipeline 11 is connected to an elbow 107, and the other end is connected to the reactor 10, and the reactor 10 is connected with a third infusion pipeline 13 (as a liquid supplement pipeline).

[0115] The starting device opens the control box 4 to generate power to drive the elastic balloon 104 to expand or contract. When the elastic balloon 104 expands, the elastic balloon 104 sucks the culture medium inside the perfusion structure, and at the same time, a negative pressure is generated inside the structure, and the culture medium containing microcarriers in the reactor 10 is sucked into the inside of the screen 106 through the first infusion pipeline 11, and the screen 106 will retain the microcarriers inside the screen 106. When the elastic balloon 104 contracts, the elastic balloon 104 spits out the culture medium inside the elastic balloon 104, flushes the screen 106, and spits out the culture medium containing microcarriers inside the screen 106 into the reactor 10, completing the exchange of the culture medium inside the perfusion structure and the culture medium in the reactor 10.

[0116] When in use, the first peristaltic pump 9 is clamped on the second infusion pipeline 12, and the culture medium outside the screen 106 is sucked out through the first peristaltic pump 9. At the same time, the culture medium without microcarriers in the perfusion structure is sucked out, and the culture medium containing microcarriers inside the screen 106 will be separated due to the pressure difference between the inside and outside of the screen 106, the microcarriers will be retained inside the screen 106, and the culture medium will pass through the screen 106 to the inside of the entire perfusion structure, and then be sucked out through the second infusion pipeline 12 by the first peristaltic pump 9. At the same time, the third infusion pipeline 13 connected to the end of the reactor 10 will supplement the liquid in the reactor 10, complete the separation of the culture medium and the microcarriers inside the reactor 10 and the update of the culture medium, and realize the replacement of the liquid in the reactor.

[0117] For the previous suspended cell perfusion structure, the elastic balloon 104 is driven to be above the entire perfusion structure, and this structure can cooperate with the sedimentation characteristics of the microcarriers (because the density of the microcarriers is greater than that of the culture medium, the microcarriers will sink in the culture medium), and the microcarriers will be concentrated near the mouth of the first infusion pipeline 11. When the elastic balloon 104 sucks, the culture medium containing microcarriers near the mouth of the main pipeline will be preferentially sucked, which is convenient for the exchange of the microcarriers inside the perfusion structure and the microcarriers inside the reactor.

[0118] For the existing microcarrier perfusion structure in the market, a small amount of bubbles will be sucked into the perfusion structure during the operation process, resulting in a large number of bubbles at the top of the perfusion structure, entering the driving diaphragm pump, and thus affecting the normal suction action of the perfusion structure.

[0119] The system drives the elastic balloon to be arranged above the entire perfusion structure, which can cooperate with the sedimentation characteristics of the microcarrier (since the microcarrier itself has a density greater than the culture medium, the microcarrier will sink in the culture medium), and the microcarrier will be concentrated near the main pipeline port. When the elastic balloon is suctioned, the culture medium with microcarriers near the main pipeline port will be preferentially suctioned, facilitating the exchange action of the microcarriers inside the perfusion structure and the microcarriers inside the reactor.

[0120] The system drives the elastic balloon to be arranged above the entire perfusion structure, which can cooperate with the sedimentation characteristics of the microcarrier (since the microcarrier itself has a density greater than the culture medium, the microcarrier will sink in the culture medium), and the microcarrier will be concentrated near the main pipeline port. When the elastic balloon is suctioned, the culture medium with microcarriers near the main pipeline port will be preferentially suctioned, facilitating the exchange action of the microcarriers inside the perfusion structure and the microcarriers inside the reactor.

[0121] The system can effectively realize the interception action of the microcarrier, complete the separation of the culture medium and the microcarrier inside the reactor and the update of the culture medium, and achieve the purpose of replacing the liquid inside the reactor. At the same time, the special structure of the system can effectively solve the problem of bubble accumulation affecting the normal perfusion action during the perfusion process, and ensure the normal operation of the microcarrier perfusion system.

[0122] The above embodiments are only used to illustrate the technical solutions of the present application, and are not limited thereto; although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that the technical solutions recorded in the foregoing embodiments can still be modified, or some or all of the technical features can be replaced by equivalents; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. An alternating tangential flow perfusion system, characterized in that, The application relates to a kind of equipment for carrying out bioreaction, which comprises: a consumable assembly (1), a weighing assembly (3), an inflation assembly, an air extraction assembly and a control box (4); the consumable assembly (1) comprises a rigid transparent spherical shell (101), an elastic balloon (104) and a hollow fiber column (105), the elastic balloon (104) is arranged in the rigid transparent spherical shell (101), the inner cavity of the elastic balloon (104) is in communication with the hollow fiber column (105) connected to the rigid transparent spherical shell (101), the butt joint of the hollow fiber column (105) is communicated with a reactor (10) through a first infusion pipeline (11), and the upper end of the hollow fiber column (105) is communicated with a waste liquid bottle (14) through a second infusion pipeline (12); the consumable assembly (1) is arranged on the weighing assembly (3), and the weighing assembly (3) is used for monitoring the weight of fluid in the consumable assembly (1) in real time; the weighing assembly (3), the inflation assembly and the air extraction assembly are connected with the control box (4), the inflation assembly is used for inflating between the elastic balloon (104) and the rigid transparent spherical shell (101), so that the elastic balloon (104) shrinks under the action of pressure difference, the air extraction assembly is used for extracting air from between the elastic balloon (104) and the rigid transparent spherical shell (101), so that the elastic balloon (104) expands under the action of pressure difference, the control box (4) judges the motion state of the elastic balloon (104) according to the change amount and the change rate of the weight of fluid, and then controls the working state of the inflation assembly and the air extraction assembly.

2. The alternating tangential flow perfusion system of claim 1, wherein, the control box (4) comprises a gas path control module, the gas path control module has an inflation path and an air extraction path, the inflation path is sequentially provided with an inflation port (609), a precision pressure regulating valve (608), a first pressure proportional valve (607), a gas flow meter (606) and a gas control electromagnetic valve (604), and the air extraction path is sequentially provided with an air extraction port (601), a vacuum control valve (602), a second pressure proportional valve (603), a gas flow meter (606) and a gas control electromagnetic valve (604).

3. The alternating tangential flow perfusion system of claim 2, wherein, the inflation assembly comprises a compressed air source and a compressed air connection pipeline (6), one end of the compressed air connection pipeline (6) is connected with the compressed air source, the other end of the compressed air connection pipeline (6) is connected with the inflation port of the control box (4), and the gas path output end of the control box (4) is connected with the consumable assembly (1) through a third connection pipeline (8).

4. The alternating tangential flow perfusion system of claim 2, wherein, the air extraction assembly comprises a vacuum pump (5) and a second connection pipeline (7), one end of the second connection pipeline (7) is connected with the vacuum pump (5), and the other end of the second connection pipeline (7) is connected with the air extraction port of the control box (4).

5. The alternating tangential flow perfusion system of claim 1, wherein, the consumable assembly (1) is further connected with a support assembly (2), the lower end of the support assembly (2) is fixed on the weighing assembly (3), and the support assembly (2) is used for supporting the consumable assembly (1).

6. The alternating tangential flow perfusion system of claim 1, wherein, The first peristaltic pump (9) is connected to the second infusion pipeline (12), the reactor (10) is communicated with the liquid supplement bottle (15) through the third infusion pipeline (13), and the second peristaltic pump (16) is connected to the third infusion pipeline (13).

7. The alternating tangential flow perfusion system of claim 1, wherein, The rigid transparent spherical shell (101) is communicated with the hollow fiber column (105) at the upper end, and the hollow fiber column (105) is provided with the second interface (109) at the upper end.

8. The alternating tangential flow perfusion system of claim 1, wherein, The consumable assembly (1) further comprises an air inlet flange (102), a screen (106) and an elbow (107). The upper end of the shell (1051) of the hollow fiber column (105) is provided with a first interface (108), and the lower end is provided with a second interface (109); the upper end of the side wall of the shell (1051) is provided with a third interface (110), and the lower end of the side wall is provided with a fourth interface (111). The rigid transparent spherical shell (101) is connected to the third interface (110) through a first chuck (112), and the other end of the rigid transparent spherical shell (101) is connected to the air inlet flange (102); the elastic balloon (104) is sleeved on the third interface (110) through an opening thereof; the screen (106) is arranged in the shell (1051), and one end of the elbow (107) is connected to the second interface (109). When the elastic balloon (104) expands under the action of pressure difference, fluid flows from the elbow (107) to the shell (1051) and flows into the elastic balloon (104) through the screen (106). When the elastic balloon (104) shrinks under the action of pressure difference, fluid flows from the elastic balloon (104) to the shell (1051) and flows back to the elbow (107) through the screen (106).

9. The alternating tangential flow perfusion system of claim 8, wherein, The rigid transparent spherical shell (101) is connected to the air inlet flange (102) through a second chuck (113), one end of the air inlet flange (102) is a chuck interface for matching the second chuck (113), and the other end is a female screw opening for connecting to a connecting pipeline (8) joint.

10. The alternating tangential flow perfusion system of claim 8, wherein, A support (103) is arranged in the elastic balloon (104), a threaded hole (114) is formed in the third interface (110), one end of the support (103) is connected to the threaded hole (114), and the other end is a flange edge for fixing the position of the elastic balloon (104).

11. The alternating tangential flow perfusion system of claim 10, wherein, A fine hole (115) is formed in the third interface (110) for communicating with the elastic balloon (104), and the fine hole (115) surrounds the threaded hole (114).

12. The alternating tangential flow perfusion system of claim 8, wherein, The elastic balloon (104) is a spherical silica gel material membrane, and the screen (106) is a stainless steel screen.

13. The alternating tangential flow perfusion system of claim 8, wherein, Both ends of the elbow (107), the first interface (108) and the second interface (109) are chuck interfaces.

14. The alternating tangential flow perfusion system of claim 8, wherein, The screen (106) has a tubular structure with one open end and the other closed end, and the open end of the screen (106) faces the elbow (107).

15. The alternating tangential flow perfusion system of claim 14, wherein, The opening end of the screen (106) and the second interface (109) are provided with a stepped surface, and the stepped surface of the screen (106) and the stepped surface of the second interface (109) are matched with each other.

16. The alternating tangential flow perfusion system of claim 14, wherein, The closed end of the screen (106) is higher than the third interface (110), and the mesh size of the screen (106) is smaller than the size of the micro carrier.

17. The alternating tangential flow perfusion system of claim 8, wherein, Further comprising: The connecting pipeline (8), the third liquid conveying pipeline (13), the fourth liquid conveying pipeline (18) and the first peristaltic pump (9); One end of the second liquid conveying pipeline (12) is connected with the first interface (108) at the upper end of the shell (1051), and the first peristaltic pump (9) is connected with the second liquid conveying pipeline (12); One end of the connecting pipeline (8) is connected with the air inlet flange (102), and the other end of the connecting pipeline (8) is connected with the control box (4); One end of the fourth liquid conveying pipeline (18) is connected with the fourth interface (111) at the lower end of the sidewall of the shell (1051); One end of the first liquid conveying pipeline (11) is connected with the elbow (107), and the other end is connected with the reactor (10), and the reactor (10) is connected with the third liquid conveying pipeline (13).

18. A method of controlling an alternating tangential flow perfusion system, characterized in that, The steps include: Step S1, inflating the rigid transparent spherical shell (101) by the inflating assembly to make the elastic balloon (104) completely shrink; Step S2, deflating the rigid transparent spherical shell (101) by the deflating assembly to make the elastic balloon (104) completely expand; Step S3, continuously operating step S1-Step S2 until the cell perfusion culture process is completed.

19. The method of controlling an alternating crossflow perfusion system according to claim 18, wherein, The step S1 includes: Step S11, in the initial state, the elastic balloon (104) is in the completely inflated state, the control box (4) controls the compressed air connecting pipeline (6) to inflate the rigid transparent spherical shell (101), so that the elastic balloon (104) gradually shrinks; in this process, the weighing assembly (3) is used to monitor the fluid weight in the consumable assembly (1) in real time; When the change amount of the fluid weight is greater than or equal to the first set value, the change rate of the fluid weight is compared with the second set value, and if the change rate of the fluid weight is less than or equal to the second set value, it is considered that the elastic balloon (104) has been completely deflated, and the switching condition is reached; Step S12, the compressed air connecting pipeline (6) is closed, and the actual operation time of the inflation operation is recorded.

20. The method of controlling an alternating crossflow perfusion system according to claim 19, wherein, Before the next inflation operation, the actual operation time of the inflation operation is compared with the theoretical operation time; If the actual operation time is greater than the theoretical operation time, the control box (4) increases the driving force of the next inflation operation through the air path control module; if the actual operation time is less than the theoretical operation time, the control box (4) reduces the driving force of the next inflation operation through the air path control module.

21. The method of controlling an alternating crossflow perfusion system of claim 18, wherein, The step S2 includes: Step S21, the control box (4) controls the vacuum pump (5) to deflate the rigid transparent spherical shell (101), so that the elastic balloon (104) gradually expands; in this process, the weighing assembly (3) is used to monitor the fluid weight in the consumable assembly (1) in real time; When the change amount of the fluid weight is greater than or equal to the third set value, the change rate of the fluid weight is compared with a fourth set value, and if the change rate of the fluid weight is less than or equal to the fourth set value, it is considered that the elastic balloon (104) has been completely inflated, and the switching condition is reached; Step S22, the vacuum pump (5) is closed, and the actual operation time of the pumping operation is recorded.

22. The method of controlling an alternating crossflow perfusion system according to claim 21, wherein, Before the next pumping operation, the actual operation time of the pumping operation is compared with the theoretical operation time; If the actual operation time is greater than the theoretical operation time, the control box (4) increases the driving force of the next pumping operation through the gas path control module; if the actual operation time is less than the theoretical operation time, the control box (4) reduces the driving force of the next pumping operation through the gas path control module.

23. The method of controlling an alternating crossflow perfusion system of claim 18, wherein, During the cyclic switching process of the inflation operation and the pumping operation, if the actual operation time exceeds the preset time and the switching condition is still not reached, the forced switching is performed according to the operation timeout logic.

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