Dual-circuit oxygen supply device, bioreactor and method

Abstract

The present application provides a dual-circuit oxygen supply device, which belongs to the field of biotechnology. The dual-circuit oxygen supply device provided by the present application comprises a first air inlet pipeline connected to a common oxygen supply device and a second air inlet pipeline connected to a backup oxygen supply device, the other ends of the two air inlet pipelines are connected to each other and to an oxygen pipeline inlet, a pressure sensor is arranged on the oxygen pipeline, an oxygen pipeline outlet is connected to a bioreactor tank body, and a mass flow meter is arranged between the oxygen pipeline outlet and the pressure sensor; a controller receives the air pressure in the oxygen pipeline detected by the pressure sensor, the opening of the mass flow meter and the current flow of the oxygen pipeline, so as to judge whether an abnormality occurs in the common oxygen supply device, and if an abnormality occurs, the controller automatically switches to the backup oxygen supply device to supply oxygen to the bioreactor, so that the oxygen supply is stable and uninterrupted, and the gas saturation in the bioreactor tank body is kept greater than a preset value, so that cells can maintain normal growth.

Description

Technical Field

[0001] The present application relates to the field of biotechnology, and in particular to a dual-path oxygen supply device, a bioreactor and a method. Background Art

[0002] When the bioreactor is in normal culture state, sterile air or oxygen needs to be continuously input into the deep layer of the culture medium in the tank for cell respiration; when the cell density increases, the input air cannot meet the cell respiration needs normally, and oxygen needs to be input to ensure normal cell respiration; if the gas supply is interrupted in the middle, it will affect the normal growth of cells and even cause cell death.

[0003] The gas supply system of the bioreactor is generally one-way air and one-way oxygen. When the cell density is low, air is supplied to ensure the normal breathing of the cells. When the cell density gradually increases, it slowly switches to supplying oxygen to ensure the normal breathing of the cells. Among them, oxygen supply equipment is generally divided into oxygen cylinders and oxygen generators. The cell culture cycle often takes several days, and the longest one takes about 30 days. Therefore, during the cell culture cycle, the oxygen cylinder has a volume limitation problem, and the oxygen generator has a certain probability of failure. Once the gas supply is interrupted in the middle, the cell state may be affected or even die.

[0004] Using oxygen generator alone to supply oxygen is difficult to avoid failure during the cell culture cycle. Using oxygen cylinders alone to supply oxygen is also difficult to ensure the endurance of gas supply over a long period of time. Especially at night when the staff are off work, the problem of oxygen supply interruption cannot be solved in time, which can easily lead to cell death and cause serious economic and time losses. Summary of the invention

[0005] The present application proposes a dual-path oxygen supply device, a bioreactor and a method for solving the problems of intermittent and insufficient stability of oxygen supply during cell culture in a bioreactor, as well as the problem of decreased gas saturation in the bioreactor tank affecting normal cell growth.

[0006] In order to achieve the above objectives, this application adopts the following scheme:

[0007] On the one hand, an embodiment of the present application provides a dual-circuit oxygen supply device, which is used in a bioreactor and includes:

[0008] An oxygen pipeline, a first air intake pipeline, a second air intake pipeline and a controller;

[0009] The first air intake pipeline is provided with a valve V1, and one end of the first air intake pipeline is connected to a common oxygen supply device;

[0010] The second air intake pipeline is provided with a valve V2, and one end of the second air intake pipeline is connected to a backup oxygen supply device;

[0011] The first air intake pipeline is connected to the other end of the second air intake pipeline and is connected to the inlet of the oxygen pipeline;

[0012] The outlet of the oxygen pipeline is connected to the tank body of the bioreactor, and a pressure sensor and a mass flow meter are arranged on the oxygen pipeline, and the mass flow meter is arranged between the pressure sensor and the outlet of the oxygen pipeline;

[0013] The pressure sensor, the mass flow meter, the valve V1 and the valve V2 are respectively connected to the controller.

[0014] Optionally, the mass flow meter comprises:

[0015] Flow control valves, flow sensors and opening controllers;

[0016] The flow control valve includes an adjustment gear and a signal receiving and sending device, and the signal receiving and sending device is connected to the opening controller;

[0017] The flow sensor and the opening controller are connected to the controller respectively.

[0018] Optionally, the controller includes:

[0019] A proportional-integral-differential-PID controller, wherein the PID controller is connected to the opening controller.

[0020] On the one hand, an embodiment of the present application provides a bioreactor, which includes any one of the dual-circuit oxygen supply devices provided in the embodiments of the present application, a reactor tank body, and a gas saturation sensor arranged inside the reactor tank body, and the gas saturation sensor is connected to the controller of the dual-circuit oxygen supply device.

[0021] Optionally, the bioreactor further comprises an air pipeline, the air pipeline inlet is connected to the air supply device, the air pipeline outlet is connected to the tank body of the bioreactor, a valve V3 is provided on the air pipeline, and the valve V3 is connected to the controller of the dual-circuit oxygen supply device.

[0022] On the one hand, an embodiment of the present application provides a dual-circuit oxygen supply method, which is used in a bioreactor provided in an embodiment of the present application, and the method comprises:

[0023] When it is first detected that the gas saturation in the bioreactor tank is less than the preset saturation, the first air inlet pipeline valve V1 is controlled to open;

[0024] Get the current opening state Q detected by the mass flow meter k ;

[0025] Get the current flow status Q of the oxygen pipeline detected by the mass flow meter y ;

[0026] When Q y ≥Q k When adjusting the mass flow meter opening Q k , until the gas saturation in the bioreactor tank is ≥ the preset saturation;

[0027] When Q y <Q k When the current pressure value P of the oxygen pipeline is obtained, if P < 1 hPa, close the first air inlet pipeline valve V1, open the second air inlet pipeline valve V2, and adjust the mass flow meter opening Q k , until the gas saturation in the bioreactor tank is ≥ the preset saturation, and a fault alarm is issued.

[0028] Optionally, the dual-path oxygen supply method further includes:

[0029] When Q y <Q k When the current pressure value P of the oxygen pipeline is obtained, if P ≥ 1 hPa, adjust the mass flow meter opening Q k , until the gas saturation in the bioreactor tank ≥ the preset saturation.

[0030] Optionally, the mass flow meter opening Q is adjusted k include:

[0031] According to the preset PID control rules, based on the current gas saturation in the bioreactor tank and the current flow state Q of the oxygen pipeline y Determine the current PID parameters;

[0032] Control the flow control valve opening Q according to the current PID parameters k .

[0033] Optionally, the gas saturation in the current bioreactor tank and the current flow state Q of the oxygen pipeline are based on the y Determining the current PID parameters includes:

[0034] According to time t, the gas saturation α in the bioreactor tank t Calculate the oxygen consumption rate v in the reactor tank at time t t ;

[0035] According to the oxygen consumption rate v t , calculate the oxygen pipeline target flow rate Q at time t t ;

[0036] According to the target flow Q t The current flow state Q of the oxygen pipeline y , determine the current PID parameters.

[0037] Optionally, the gas saturation in the current bioreactor tank and the current flow state Q of the oxygen pipeline are based on the y Determining the current PID parameters also includes:

[0038] According to the cell culture type carried out in the bioreactor tank at time t, and the oxygen consumption rate v in the bioreactor tank t , predict the current reaction stage;

[0039] According to the current reaction stage and the oxygen consumption rate v in the bioreactor tank t , predict the oxygen consumption rate v in the bioreactor tank at time t+1 t+1 ;

[0040] According to the oxygen consumption rate v in the bioreactor tank t+1 , adjust the current PID parameters.

[0041] The beneficial effects brought by the technical solution provided by the embodiment of the present application include at least:

[0042] The dual-circuit oxygen supply device provided by the present application includes a first air intake pipeline connected to a common oxygen supply device and a second air intake pipeline connected to a backup oxygen supply device, the other ends of the two air intake pipelines are connected to each other and to an oxygen pipeline inlet, a pressure sensor is provided on the oxygen pipeline, the oxygen pipeline outlet is connected to a bioreactor tank body, and a mass flow meter is provided between the oxygen pipeline outlet and the pressure sensor; the controller receives the air pressure in the oxygen pipeline detected by the pressure sensor, the opening of the mass flow meter and the current flow of the oxygen pipeline, so as to determine whether the common oxygen supply device has an abnormality, and if an abnormality occurs, the backup oxygen supply device is switched to supply oxygen to the bioreactor, so that the oxygen supply is stable and uninterrupted, and the gas saturation in the bioreactor tank body is maintained. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 A schematic diagram of the structure of a dual-circuit oxygen supply device provided as an exemplary embodiment of the present application;

[0044] Figure 2 A schematic diagram of the structure of a mass flow meter for a dual-path oxygen supply device provided by an exemplary embodiment of the present application;

[0045] Figure 3 A schematic diagram of a bioreactor structure provided for an exemplary embodiment of the present application;

[0046] Figure 4 A flow chart of a dual-path oxygen supply method provided for an exemplary embodiment of the present application;

[0047] Icon Description:

[0048] 1 oxygen pipeline; 1.1 first air intake pipeline; 1.2 second air intake pipeline;

[0049] 2. Pressure sensor;

[0050] 3Mass flow meter; 3.1Flow sensor; 3.2Flow control valve; 3.3Opening controller;

[0051] 4 controller; 5 reactor tank; 6 gas saturation sensor; 7 air pipeline. DETAILED DESCRIPTION

[0052] In order to make the purpose, technical scheme and advantages of the embodiment of the present application clearer, the technical scheme of the embodiment of the present application will be clearly and completely described below in conjunction with the drawings of the embodiment of the present application. Obviously, the described embodiment is a part of the embodiment of the present application, not all of the embodiments. Based on the described embodiment of the present application, all other embodiments obtained by ordinary technicians in this field without making creative work belong to the scope of protection of the present application.

[0053] In the description of this application, it should be noted that if the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc. appear, the orientation or position relationship indicated is based on the orientation or position relationship shown in the drawings, or the orientation or position relationship in which the product of the application is usually placed when in use. It is only for the convenience of describing this application and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as a limitation on this application. In addition, if the terms "first", "second", etc. appear in the description of this application, they are only used to distinguish the description and cannot be understood as indicating or implying relative importance.

[0054] In the description of this application, it should also be noted that, unless otherwise clearly specified and limited, the terms "set", "install", "connect", and "connect" should be understood in a broad sense. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal connection of two components. For ordinary technicians in this field, the specific meanings of the above terms in this application can be understood according to specific circumstances.

[0055] In addition, the technical features involved in the different embodiments of the present application described below can be combined with each other as long as they do not conflict with each other.

[0056] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0057] Figure 1 A dual-circuit oxygen supply device provided in an embodiment of the present application is shown, and the dual-circuit oxygen supply device is used in a bioreactor, see Figure 1 ,include:

[0058] Oxygen pipeline 1, first air intake pipeline 1.1, second air intake pipeline 1.2 and controller 4;

[0059] A valve V1 is provided on the first air intake pipeline 1.1, and one end of the first air intake pipeline 1.1 is connected to a common oxygen supply device;

[0060] A valve V2 is provided on the second air intake pipeline 1.2, and one end of the second air intake pipeline 1.2 is connected to the backup oxygen supply device;

[0061] The first air intake pipeline 1.1 is connected to the other end of the second air intake pipeline 1.2 and is connected to the inlet of the oxygen pipeline 1;

[0062] The outlet of the oxygen pipeline 1 is connected to the tank body of the bioreactor, and a pressure sensor 2 and a mass flow meter 3 are arranged on the oxygen pipeline 1, and the mass flow meter 3 is arranged between the pressure sensor 2 and the outlet of the oxygen pipeline 1;

[0063] The pressure sensor 2 , the mass flow meter 3 , the valve V1 and the valve V2 are respectively connected to the controller 4 .

[0064] Exemplarily, in some embodiments, the common oxygen supply device connected to one end of the first air intake pipeline 1.1 may be an oxygen cylinder or an oxygen concentrator; the backup oxygen supply device connected to one end of the second air intake pipeline 1.2 may be an oxygen cylinder.

[0065] For example, in some embodiments, the valve V1 and the valve V2 may be solenoid valves, and the opening and closing of the valve V1 and the valve V2 may be controlled by the controller 4 .

[0066] For example, in some embodiments, the controller 4 may use a programmable logic controller (PLC).

[0067] Exemplarily, the dual-circuit oxygen supply device has two air inlet pipelines, which are respectively connected to a conventional oxygen supply device and a backup oxygen supply device. The air pressure state of the oxygen pipeline is detected by a pressure sensor, and the flow state of the oxygen pipeline near the outlet end is detected by a mass flow meter. The controller determines whether there is an abnormality in the conventional oxygen supply device based on the information monitored by the pressure sensor and the mass flow meter. If an abnormality occurs, the controller closes valve V1 and opens valve V2 to switch to the backup oxygen supply device to supply oxygen to the bioreactor, and at the same time, an alarm is issued for abnormalities in the conventional oxygen supply device.

[0068] The above-mentioned oxygen supply device is connected to the common oxygen supply device and the backup oxygen supply device through two air inlet pipelines respectively. Combined with the real-time detection of the pressure sensor and the mass flow meter, the controller generates control instructions by receiving the detection information and sends them to related equipment, which can realize continuous and stable oxygen supply to the bioreactor, and maintain the stability of the oxygen saturation in the reactor tank, reducing the probability of cell culture failure due to oxygen supply problems; at the same time, there is a backup oxygen supply device and pipeline, and there is no need to arrange personnel to be on duty at night, or to manually pay long-term attention to whether there are problems with the oxygen supply end; the oxygen supply device leaves maintenance personnel with enough time to deal with problems.

[0069] Optional, see Figure 2 , the mass flow meter 3 comprises:

[0070] Flow sensor 3.1, flow control valve 3.2 and opening controller 3.3;

[0071] The flow sensor 3.1 and the opening controller 3.3 are respectively connected to the controller;

[0072] The flow control valve 3.2 includes an adjustment gear and a signal receiving and sending device, and the signal receiving and sending device is connected to the opening controller 3.3.

[0073] For example, Figure 2 As shown, the flow sensor 3.1 is connected to the controller 4, and sends the detected gas flow in the oxygen pipeline to the controller 4 in real time; the opening controller 3.3 obtains the status information of the flow control valve 3.2, and sends the status information of the flow control valve 3.2 to the controller 4, and the opening controller 3.3 obtains the instruction information of the controller 4, and sends it to the flow control valve 3.2 to adjust the opening of the mass flow meter.

[0074] The flow sensor in the mass flow meter can detect the current gas flow in the oxygen pipeline, and the flow control valve can control the opening of the current metering flow meter. The current opening state is sent and received through the opening controller, and the opening state of the mass flow meter is adjusted by receiving instructions from the controller, so that the opening state of the mass flow meter can be adjusted in real time to make the oxygen supply more stable, thereby stabilizing the gas saturation in the tank of the bioreactor.

[0075] Optionally, the controller 4 includes:

[0076] Proportional-integral-differential-PID controller, the PID controller is connected to the opening controller 3.3.

[0077] Exemplarily, the controller 4 includes a PID controller, which analyzes, processes and calculates the information received by the controller 4 to generate PID parameters, which are sent to the opening controller 3.3. The opening controller 3.3 sends an opening adjustment instruction to control the opening adjustment of the flow control valve 3.2.

[0078] Using a proportional-integral-differential PID controller to adjust the opening of the flow control valve of the mass flow meter can make the flow control more accurate, meet the oxygen demand in the bioreactor tank, and maintain stable gas saturation.

[0079] Figure 3 It is shown that an embodiment of the present application provides a bioreactor, which includes any one of the dual-circuit oxygen supply devices provided in the embodiments of the present application, a reactor tank body 5, and a gas saturation sensor 6 arranged inside the reactor tank body, and the gas saturation sensor 6 is connected to the controller 4.

[0080] For example, Figure 3 As shown, the outlet of the oxygen pipeline 1 of the dual-circuit oxygen supply device is connected to the reactor tank body 5, and a gas saturation sensor 6 is provided in the reactor tank body 5. The controller 4 can receive monitoring information from the gas saturation sensor 6, so as to adjust the dual-circuit oxygen supply device in time according to the gas saturation in the reactor tank body.

[0081] Optionally, the bioreactor further comprises an air pipeline 7, the air pipeline inlet is connected to the air supply device, the air pipeline outlet is connected to the tank body of the bioreactor, a valve V3 is arranged on the air pipeline, and the valve V3 is connected to the controller of the dual-circuit oxygen supply device.

[0082] The controller can control the first air inlet pipeline valve V1, the second air inlet pipeline valve V2 and the air pipeline valve V3. By controlling the above three valves, gas is supplied to the bioreactor tank to ensure the stability of the gas saturation in the reactor tank.

[0083] Combine the following Figure 1 , Figure 2 and Figure 3 The specific use of the dual-circuit oxygen supply device provided by the present application is further described by the above dual-circuit feeding device and the bioreactor:

[0084] Place the cells to be cultured and the culture fluid into the reactor tank 5, determine the gas saturation required for the cell culture type according to the preset, close the reactor tank 5, and start the cell culture;

[0085] It is worth noting that the gas saturation here refers to the oxygen saturation in the reactor tank. For example, if the proportion of oxygen in the air is 21%, only air is input into the reactor tank when the reaction starts. At this time, the oxygen pipeline is closed. As the reaction proceeds, oxygen is continuously consumed and the oxygen content in the reactor decreases. In order to facilitate the adjustment and measurement of the control parameters, the gas saturation sensor 6 uses 21% oxygen content in the air as the state of 100% gas saturation in the reactor tank.

[0086] The controller 4 controls the valve V3 on the air pipeline 7 to open and supply air to the reactor tank 5. As the reaction proceeds, the oxygen consumed by the cells gradually increases, and the air supply alone cannot meet the needs of cell culture. The gas saturation in the reactor tank 5 decreases from the initial 100% to a preset value (such as the preset gas saturation in the reactor tank is >40%). If the gas saturation in the reactor tank 5 is 40% at this time, the controller 4 controls V1 to open, and adjusts the opening of the mass flowmeter 3 according to the current flow and opening state detected by the pressure sensor 2 and the mass flowmeter 3, and the oxygen consumption rate in the reactor tank 5, so that the gas saturation in the reactor tank 5 is maintained at >40%.

[0087] The pressure sensor 2 and the mass flow meter 3 maintain real-time detection. When it is detected that the current flow rate is less than the current opening state and the current air pressure in the oxygen pipeline is less than 1 bar (the unit of air pressure: hPa), the controller closes the valve V1, opens the valve V2, adjusts the opening of the mass flow meter to keep the gas saturation in the reactor tank greater than 40%, and issues a fault alarm for the common oxygen pipeline; if it is detected that the current flow rate is less than the current opening state and the current air pressure in the oxygen pipeline is ≥1 bar, the controller does not adjust the valve V1 and / or valve V2, and only adjusts the opening of the mass flow meter to keep the gas saturation in the reactor tank greater than 40%.

[0088] Figure 4 The figure shows a schematic flow diagram of a dual-path oxygen supply method provided in an embodiment of the present application, which is used in a bioreactor provided in an embodiment of the present application, and the method comprises:

[0089] Step 401, when it is first detected that the gas saturation in the bioreactor tank is less than the preset saturation, the first air inlet pipeline valve V1 is controlled to open;

[0090] The gas saturation in the bioreactor tank is acquired by a gas saturation sensor arranged in the bioreactor tank and sent to the controller, and the first air inlet pipeline valve V1 is controlled by the controller;

[0091] Step 402, obtaining the current opening state Q detected by the mass flow meter k ;

[0092] Among them, the current opening state Q k It is acquired by the opening controller in the mass flow meter and sent to the controller;

[0093] Step 403, obtaining the current flow state Q of the oxygen pipeline detected by the mass flow meter y ;

[0094] Among them, the current flow state of the oxygen pipeline is Q y It is acquired by the flow sensor in the mass flow meter and sent to the controller;

[0095] Step 404, determine whether: the current flow state Q y <Current opening state Q k ; If yes, proceed to step 405, otherwise proceed to step 406;

[0096] Step 405, obtaining the current pressure value P of the oxygen pipeline, and proceeding to step 407;

[0097] Among them, the current air pressure value P of the oxygen pipeline is obtained by the pressure sensor on the oxygen pipeline and sent to the controller;

[0098] Step 406, adjust the mass flow meter opening Q k , until the gas saturation in the bioreactor tank is ≥ the preset saturation;

[0099] The controller adjusts the mass flow meter opening Q according to the preset and received information. k Make adjustments to keep the gas saturation in the bioreactor tank ≥ the preset saturation;

[0100] Step 407, determine whether the current pressure value P of the oxygen pipeline is less than 1 bar; if yes, proceed to step 408, otherwise proceed to step 406;

[0101] Step 408, close the first air intake pipeline valve V1, open the second air intake pipeline valve V2, jump to step 406, step 409;

[0102] It is worth noting that the current traffic state Q y <Current opening state Q k When saving the following:

[0103] One is that valve V1 or valve V2 has just been opened, and oxygen has not yet filled the entire oxygen pipeline. At this time, the opening of the mass flow meter will be greater than the current flow rate of the oxygen pipeline. Therefore, in this case, it is necessary to detect the air pressure at the entrance of the oxygen pipeline. If the front air pressure value P ≥ 1 bar, there is no fault in the air inlet pipeline, and there is no need to adjust the state of valve V1 or valve V2. It is only necessary to wait for oxygen to fill the entire oxygen pipeline and adjust the opening of the mass flow meter so that the gas saturation in the bioreactor tank can be maintained ≥ the preset saturation;

[0104] Another situation is that the common oxygen supply device fails and cannot supply gas normally through the first air inlet pipeline. At this time, by detecting the air pressure at the entrance of the oxygen pipeline, if the front air pressure value P is less than 1 bar, the controller confirms that the common oxygen supply device fails and needs to switch to the backup oxygen supply device for oxygen supply. At this time, the controller controls valve V1 to close, valve V2 to open, and adjusts the opening of the mass flow meter to keep the gas saturation in the bioreactor tank ≥ the preset saturation;

[0105] Step 409, fault alarm is issued;

[0106] The fault alarm is controlled by the controller, and the controller may give an alarm prompt through an alarm device connected thereto, or a monitoring station or other equipment.

[0107] Repeat the above steps to keep the gas saturation in the bioreactor tank ≥ the preset saturation.

[0108] Optionally, adjust the mass flow meter opening Q k include:

[0109] According to the preset PID control rules, based on the current gas saturation in the bioreactor tank and the current flow state Q of the oxygen pipeline y Determine the current PID parameters;

[0110] Control the flow control valve opening Q according to the current PID parameters k .

[0111] For example, the PID control rules are obtained by the staff through multiple tests and debugging according to different cell culture requirements, and then preset in the controller. When cell culture is carried out, it can be based on the current gas saturation in the bioreactor tank and the current flow state Q of the oxygen pipeline. y Determine the PID parameters suitable for the current adjustment.

[0112] PID control can make the gas saturation in the bioreactor tank more stable. By obtaining the corresponding PID control rules through multiple tests and debugging and presetting them in the controller, it can simplify the operation, standardize the process, reduce the workload of staff, and improve work efficiency and the stability of cell culture during actual cell culture.

[0113] Optionally, based on the current gas saturation in the bioreactor tank and the current flow state Q of the oxygen pipeline y Determining the current PID parameters includes:

[0114] According to the gas saturation α in the bioreactor tank at time t, t Calculate the oxygen consumption rate v in the reactor tank at time t t ;

[0115] According to the oxygen consumption rate v t , calculate the oxygen pipeline target flow rate Q at time t t ;

[0116] According to the target flow Q t The current flow status Q of the oxygen pipeline y , determine the current PID parameters.

[0117] Exemplarily, the controller can be configured to calculate the oxygen consumption rate v in the reactor tank at time t. t Calculate how much oxygen flow the current oxygen pipeline needs to provide to meet the current rate of oxygen consumption by cells in the bioreactor tank. This oxygen flow is the target flow Q t , according to the target flow Q t The current flow status Q of the oxygen pipeline y The difference can be used to adjust the mass flow meter opening state Q k During the process, the target flow is constantly corrected through PID control, thereby adjusting the PID parameters so that the current flow state Q y Approach target flow rate Q more quickly and accurately t ;

[0118] It is worth noting that as the cell culture progresses, the rate at which cells consume oxygen is constantly changing, so the target flow rate Q t It will change over time, so the PID parameters need to be constantly updated and corrected to keep the oxygen saturation in the bioreactor tank greater than or equal to the preset value. Therefore, the dual-path oxygen supply method provided in this application adopts PID control in order to continuously correct the target flow rate Q t , to achieve better control effect.

[0119] Optionally, based on the current gas saturation in the bioreactor tank and the current flow state Q of the oxygen pipeline y Determining the current PID parameters also includes:

[0120] According to the type of cell culture carried out in the bioreactor tank at time t, and the oxygen consumption rate v in the bioreactor tank t , predict the current reaction stage;

[0121] According to the current reaction stage and the oxygen consumption rate v in the bioreactor tank t , predict the oxygen consumption rate v in the bioreactor tank at time t+1 t+1 ;

[0122] According to the oxygen consumption rate v in the bioreactor tank t+1 , adjust the current PID parameters.

[0123] For example, if the cell culture type in the current bioreactor tank is A cells, and the culture period is 30 days, the relationship between the oxygen consumption rate v and the culture process during cell culture in the bioreactor provided in the present application is v(t)=m a q(m at ), where t is the time when the reaction is carried out, m a is the initial amount of cultured A cells, q(m at ) is the function of oxygen consumption at time t when the initial amount of A cells is m during cultivation;

[0124] At a certain point on the tenth day of the reaction, the oxygen consumption rate in the reactor is v t10 According to the function of oxygen consumption of cell A at the current reaction moment, the controller can estimate the reaction progress and predict the oxygen consumption at the next moment, so as to adjust the current PID parameters so that the gas saturation in the bioreactor tank can be kept greater than or equal to the preset value, and the gas saturation will not drop below the preset value due to the delay in adjusting the mass flow meter during the reaction stage, thus affecting the normal growth of cells.

[0125] The dual-path oxygen supply method provided in the embodiment of the present application can be applied to the bioreactor provided in the embodiment of the present application. From the beginning of the reaction, the gas saturation in the bioreactor tank is monitored in real time. When the gas saturation in the bioreactor tank is detected to be less than a preset value for the first time, the control valve V1 is opened to supply oxygen to the reactor tank. When supplying oxygen, the current opening state Q of the mass flow meter is monitored in real time. k The current flow status Q of the oxygen pipeline y , when Q y <Q kWhen the oxygen pipeline pressure is less than 1 bar, the controller confirms that the common gas supply device is abnormal, closes valve V1, opens valve V2, and issues a fault alarm; in other cases, the status of valves V1 and V2 is not adjusted;

[0126] The dual-circuit oxygen supply method provided in the embodiment of the present application can automatically adjust the oxygen supply status in the bioreactor. When the normal oxygen supply device is abnormal, it automatically switches to the backup oxygen supply device and issues an alarm to remind the staff to maintain the oxygen supply equipment. The gas saturation in the reactor tank and the status of the dual-circuit oxygen supply device are monitored in real time. The gas saturation in the bioreactor tank is kept stable and greater than or equal to the preset value through PID control, so that the cells can grow normally and avoid the problem of culture failure caused by unstable or interrupted oxygen supply.

[0127] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection created by this application.

Claims

1. A bioreactor, characterized in that: It includes a dual-path oxygen supply device, a reactor tank body, and a gas saturation sensor arranged inside the reactor tank body; The dual-circuit oxygen supply device comprises: An oxygen pipeline, a first air intake pipeline, a second air intake pipeline and a controller; The first air intake pipeline is provided with a valve V1, and one end of the first air intake pipeline is connected to a common oxygen supply device, wherein the common oxygen supply device is an oxygen cylinder or an oxygen concentrator; The second air intake pipeline is provided with a valve V2, and one end of the second air intake pipeline is connected to a backup oxygen supply device, wherein the backup oxygen supply device is an oxygen cylinder; The first air intake pipeline is connected to the other end of the second air intake pipeline and is connected to the inlet of the oxygen pipeline; The outlet of the oxygen pipeline is connected to the tank body of the bioreactor, and a pressure sensor and a mass flow meter are arranged on the oxygen pipeline, and the mass flow meter is arranged between the pressure sensor and the outlet of the oxygen pipeline; The pressure sensor, the mass flow meter, the valve V1 and the valve V2 are respectively connected to the controller; The mass flow meter comprises: Flow control valves, flow sensors and opening controllers; The flow control valve includes an adjustment gear and a signal receiving and sending device, and the signal receiving and sending device is connected to the opening controller; The flow sensor and the opening controller are respectively connected to the controller; The controller comprises: a proportional-integral-differential-integral PID controller, wherein the PID controller is connected to the opening controller; The gas saturation sensor is connected to the controller of the dual-path oxygen supply device.

2. The bioreactor according to claim 1, characterized in that The bioreactor also includes an air pipeline, on which a valve V3 is provided, the air pipeline inlet is connected to the air supply device, the air pipeline outlet is connected to the tank body of the bioreactor, and the valve V3 is connected to the controller of the dual-circuit oxygen supply device.

3. A dual-circuit oxygen supply method, the dual-circuit oxygen supply method is used for the bioreactor as claimed in claim 1 or 2, characterized in that: The method comprises: When it is detected for the first time that the gas saturation in the bioreactor tank is less than the preset saturation, the first air inlet pipeline valve V1 is controlled to open; Get the current opening state Q detected by the mass flow meter k ; Get the current flow status Q of the oxygen pipeline detected by the mass flow meter y ; When Q y ≥Q k When adjusting the mass flow meter opening Q k , until the gas saturation in the bioreactor tank is ≥ the preset saturation; When Q y <Q k When the current pressure value P of the oxygen pipeline is obtained, if P < 1 hPa, close the first air inlet pipeline valve V1, open the second air inlet pipeline valve V2, and adjust the mass flow meter opening Q k , until the gas saturation in the bioreactor tank is ≥ the preset saturation, and a fault alarm is issued.

4. The dual-path oxygen supply method according to claim 3, characterized in that: The dual-path oxygen supply method further comprises: When Q y <Q k When the current pressure value P of the oxygen pipeline is obtained, if P ≥ 1 hPa, adjust the mass flow meter opening Q k , until the gas saturation in the bioreactor tank ≥ the preset saturation.

5. The dual-path oxygen supply method according to claim 3, characterized in that: The mass flow meter opening Q is adjusted k include: According to the preset PID control rules, based on the current gas saturation in the bioreactor tank and the current flow state Q of the oxygen pipeline y Determine the current PID parameters; Control the flow control valve opening Q according to the current PID parameters k .

6. The dual-path oxygen supply method according to claim 5, characterized in that: The method is based on the gas saturation in the current bioreactor tank and the current flow state Q of the oxygen pipeline y Determining the current PID parameters includes: According to time t, the gas saturation α in the bioreactor tank t Calculate the oxygen consumption rate v in the reactor tank at time t t ; According to the oxygen consumption rate v t , calculate the oxygen pipeline target flow rate Q at time t t ; According to the target flow Q t The current flow state Q of the oxygen pipeline y , determine the current PID parameters.

7. The dual-path oxygen supply method according to claim 6, characterized in that: The method is based on the gas saturation in the current bioreactor tank and the current flow state Q of the oxygen pipeline y Determining the current PID parameters also includes: According to the cell culture type carried out in the bioreactor tank at time t, and the oxygen consumption rate v in the bioreactor tank t , predict the current reaction stage; According to the current reaction stage and the oxygen consumption rate v in the bioreactor tank t , predict the oxygen consumption rate v in the bioreactor tank at time t+1 t+1 ; According to the oxygen consumption rate v in the bioreactor tank t+1 , adjust the current PID parameters.

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