An automated control system and method for microbial growth

Abstract

The present application belongs to the technical field of bioengineering, and relates to an automatic control system and method for microbial growth. The method comprises the following steps: providing a biological reaction system for microbial growth, wherein the reaction system comprises a bioreactor and a supporting device for supporting the bioreactor; providing a monitoring module for monitoring working state parameters of the bioreactor; introducing gas or liquid into a reaction bag through an inlet to perform biological culture in response to a corresponding control signal sent by a user or the monitoring module; monitoring the liquid in a second space to collect a hydraulic pressure and monitoring the bottom of the reaction bag to collect a liquid leakage value; and judging a risk state of the biological reaction system according to the hydraulic pressure and the liquid leakage value. The automatic control method for microbial growth provided by the present application can automatically monitor and sample the whole process of microbial culture, and can regulate the safety and stability of the biological culture process through risk monitoring.

Description

Technical Field

[0001] This invention belongs to the field of bioengineering technology, specifically relating to an automated control system and method for microbial growth. Background Technology

[0002] Single-use bioreactors are a novel type of bioreactor, typically made of disposable plastic or stainless steel. These bioreactors are commonly used in fields such as cell culture, bacterial culture, virus culture, and the cultivation and extraction of effective substances like monoclonal antibodies. However, existing bioreactors are generally used in large-scale cell or microbial cultures (such as commercial applications) and are insufficient for small-scale experiments. For example, in the early stages of vaccine development, researchers often need to conduct multiple small-scale virus cultures in the laboratory. However, existing biosafety reactors have limitations in terms of cost, safety, and ease of operation for small-scale virus culture.

[0003] For example, Chinese invention patent CN102492607B discloses a disposable bioreactor system and method. This disposable bioreactor system includes: a disposable container with at least one inlet, at least one outlet, and at least one collection port, using sterile filters connected to all external openings to maintain the integrity of the sterile environment; a structure supporting the disposable container; one or more sensors for sensing one or more parameters of the biological material in the container; a heater for heating the contents of the container, the heater having a thermostat; and a mixing system integrated with the system to mix the biological material contained in the disposable container. This bioreactor system is relatively large in size, employing a large-capacity bioreactor and requiring a series of auxiliary equipment, resulting in high costs and greater demands on experimental operating space. Furthermore, there are significant differences between large-scale microbial or cell culture and micro-microbial or cell culture in practical applications. Therefore, even proportionally scaling down existing equipment is insufficient to meet the needs of micro-scale experiments in laboratories. For example, Chinese invention patent application CN103945928A discloses a single-use mixing and bioreactor system, which will also have similar problems when applied to micro-scale experiments.

[0004] To further adapt to the needs of micro-scale experiments, some small-scale reactor designs have been proposed in the prior art. For example, Chinese patent application CN102212474A discloses a small bioreactor comprising five parts: a cell culture tank, a temperature control kit, a liquid (gas) replenishment device, a cell optical density detection kit, and a culture control device. The cell culture tank is connected to the liquid replenishment device, and the temperature control kit, cell optical density detection kit, and culture control device work together to provide optimized growth conditions for cell culture. However, this type of small bioreactor requires steam sterilization after each reaction, which is cumbersome and costly for batch experiments. Furthermore, when used to culture pathogenic microorganisms such as bacteria or viruses, it is difficult to ensure that the internal and external environments of the reactor are not disturbed (for example, pathogenic bacteria in the reactor may spill out and contaminate the experimental environment, or even threaten the safety of operators).

[0005] Therefore, there is an urgent need for a bioreactor and system suitable for small-scale microbial or cell culture scenarios. Summary of the Invention

[0006] The purpose of this invention is to provide a disposable biosafety bioreactor and monitoring method to partially solve or alleviate the above-mentioned deficiencies in the prior art, and to effectively solve the difficulties in the miniaturized culture process of cells or microorganisms.

[0007] To address the aforementioned technical problems, the present invention specifically adopts the following technical solution: an automated control method for microbial growth, comprising:

[0008] S301 provides a bioreactor system for microbial growth, the bioreactor system comprising: a bioreactor,

[0009] And a support device for supporting the bioreactor, wherein the bioreactor includes: a reaction bag, the inside of which includes a first space and a second space for containing gas and liquid respectively, and the reaction bag is also provided with a first inlet and a second inlet for introducing gas and liquid respectively, and a working status monitoring module for monitoring the working status parameters of the bioreactor;

[0010] S302 responds to a corresponding control signal issued by the user or the working status monitoring module through the first input and / or

[0011] Alternatively, a second inlet may be used to introduce gas and / or liquid into the reaction bag for biological culture.

[0012] S303 performs pressure monitoring on the liquid inside the second space to collect hydraulic pressure, and performs leakage monitoring on the bottom of the reaction bag to collect leakage value;

[0013] S304 determines the risk status of the bioreactor system based on the hydraulic pressure and the leakage value. The risk status includes: a safe status, and / or a pending status, and / or a leakage status. S304 includes: determining the pressure reduction risk of the reaction bag based on the changing parameters of the hydraulic pressure. The pressure reduction risk is divided into three levels based on the magnitude of the changing parameters: level zero, level one, and level two.

[0014] The leakage risk of the reaction bag is determined based on the leakage value; wherein the leakage risk is divided into three levels according to the magnitude of the leakage value: level zero, level one, and level two.

[0015] The risk status is determined based on the pressure reduction risk and leakage risk; wherein, when the pressure reduction risk is level two and / or...

[0016] When the leakage risk is level 2, the risk status is leakage status; when both the pressure drop risk and the leakage risk are level 0, the risk status is safe status; when the pressure drop risk is level 1 and the leakage risk is level 0, or when the pressure drop risk is level 0 and the leakage risk is level 1, the risk status is pending status.

[0017] In some embodiments, it also includes:

[0018] When S305 detects that the risk status is either pending or leaking, it will issue a corresponding warning signal to the user.

[0019] S306 responds to the feedback signal sent by the user by performing secondary monitoring of the pressure reduction risk and / or leakage risk of the bioreactor system, so as to receive or correct the warning signal.

[0020] In some embodiments, a condensate guiding zone is further formed in the top region of the first space. The condensate guiding zone is used to guide condensate to condense within the guiding zone and to flow back to the second space through the guiding path provided by the condensate guiding zone. A heating unit is also provided in the condensate guiding zone. Accordingly, the feedback signal includes: a first feedback signal for heating the condensate guiding zone, and a second feedback signal for receiving the warning signal. S306 includes the following steps:

[0021] When the pressure drop risk is level one and the leakage risk is level zero, the heating unit responds to the first feedback signal to heat the condensate guide area;

[0022] When the pressure drop risk is detected to drop to zero during or after heating, the risk state is corrected to a safe state in response to a first correction signal issued by the user; otherwise, the risk state is corrected to a leakage state in response to a second correction signal issued by the user.

[0023] In some embodiments, S306 further includes the step of:

[0024] When the pressure drop risk is detected to be level zero or the leakage risk is level one, the leakage value is monitored. When the leakage value is detected to remain constant within a preset fourth time period, the risk state is corrected to a safe state in response to the first correction signal; otherwise, the risk state is corrected to a leakage state in response to the second correction signal.

[0025] In some embodiments, the bioreactor system further includes: a stirring device, the stirring device including a stirring shaft and a stirring paddle disposed at the end of the stirring shaft; a mounting hole for mounting the stirring device is provided in the central region of the condensate guiding zone, and the stirring shaft is fixedly mounted on the reaction bag through the mounting hole; the heating unit is disposed on the side of the stirring shaft located at or near the condensate guiding zone, for avoiding or limiting the formation of condensate.

[0026] In some embodiments, the wall of the reaction bag inside the condensate guiding zone is configured as a slope, and the slope angle is 1°-5°.

[0027] In some embodiments, the bioreactor system further includes a safety disinfection module for disinfecting the bioreactor and the support device, the safety disinfection module including a recovery unit connected to the reaction bag via a recovery pipe, and a chamber for storing disinfectant, the chamber being connected to the reaction bag and the support device via disinfection pipes respectively; the method further includes the step of:

[0028] When S307 detects that the risk state is a leakage state, it sends a corresponding disinfection signal to the safety disinfection module.

[0029] S308 The safety disinfection module responds to the disinfection signal by recovering the liquid inside the reaction bag to the recovery unit, and closes the recovery pipeline after the recovery is completed;

[0030] S309 After the recycling pipeline is closed, the safety disinfection module responds to the disinfection signal by introducing the disinfectant into the reaction bag and the support device.

[0031] In some embodiments, the step prior to 301 is further included:

[0032] S300 performs a preliminary stability test on the bioreactor system; wherein, S300 includes:

[0033] Gas is introduced into the reaction bag through the first inlet, and the second gas pressure change value inside the support device is collected simultaneously; at the same time, the other inlets and outlets of the reaction bag and the support device are kept closed.

[0034] End ventilation and keep the corresponding first inlet closed;

[0035] The first pressure change value of the reaction bag within a preset first time period and the third pressure change value of the support device within the first time period are collected.

[0036] The stability of the bioreactor system is determined based on the first, second, and third pressure change values; among which...

[0037] When the first pressure change value is within the preset corresponding safety threshold, and the second and third pressure change values ​​are consistent with the amount of gas introduced, the bioreactor system passes the pre-test.

[0038] The present invention also provides an automated control system for microbial growth, comprising:

[0039] A bioreactor system, comprising: a bioreactor, and a support device for supporting the bioreactor, wherein the bioreactor comprises: a reaction bag, the reaction bag having a first space and a second space for containing gas and liquid respectively, and the reaction bag also having a first inlet and a second inlet for introducing gas and liquid respectively, and a working status monitoring module for monitoring the working status parameters of the bioreactor;

[0040] The sample addition module is configured to introduce gas and / or liquid into the reaction bag through the first inlet and / or the second inlet for biological culture in response to a corresponding control signal issued by the user or the working status monitoring module.

[0041] A leakage detection module is configured to monitor the pressure of the liquid inside the second space to collect hydraulic data.

[0042] Leakage monitoring is performed at the bottom of the reaction bag to collect leakage values;

[0043] A risk analysis module is configured to determine the risk status of the bioreactor system based on the hydraulic pressure and the leakage value, the risk status including: a safe status, and / or a pending status, and / or a leakage status; wherein the risk analysis module includes:

[0044] A hydraulic analysis unit is configured to determine the risk of depressurization of the reaction bag based on changes in the hydraulic pressure parameters.

[0045] The risk of blood pressure reduction is classified into three levels based on the magnitude of the changing parameters: Level 0, Level 1, and Level 2.

[0046] A leakage analysis unit is configured to determine the leakage risk of the reaction bag based on the leakage value; wherein the leakage risk is classified into three levels according to the magnitude of the leakage value: level zero, level one, and level two.

[0047] The risk analysis unit is configured to determine the risk status based on the pressure drop risk and leakage risk; wherein,

[0048] When the pressure drop risk is level 2 and / or the leakage risk is level 2, the risk status is a leakage status; when both the pressure drop risk and the leakage risk are level 0, the risk status is a safe status; when the pressure drop risk is level 1 and the leakage risk is level 0, or when the pressure drop risk is level 0 and the leakage risk is level 1, the risk status is a pending status.

[0049] In some embodiments, it also includes:

[0050] The early warning sending module is configured to send an alert when the detected risk status is either pending or leaking.

[0051] Send corresponding warning signals to users;

[0052] The early warning correction module is configured to perform secondary monitoring of the pressure reduction risk and / or leakage risk of the bioreactor system in response to feedback signals issued by the user, so as to receive or correct the early warning signals.

[0053] In some embodiments, a condensate guiding zone is further formed in the top region of the first space. The condensate guiding zone is used to guide condensate to condense within the guiding zone and to flow back to the second space through the guiding path provided by the condensate guiding zone. Accordingly, the feedback signal includes: a first feedback signal for heating the condensate guiding zone, and a second feedback signal for receiving the warning signal. The warning correction module includes:

[0054] The first correction unit is configured to heat the condensate guide zone in response to a received first feedback signal when the pressure drop risk is level one and the leakage risk is level zero; and when it is detected that the pressure drop risk drops to level zero during or after heating, the risk state is corrected to a safe state in response to a first correction signal issued by the user; otherwise, the risk state is corrected to a leakage state in response to a second correction signal issued by the user.

[0055] Beneficial technical effects:

[0056] This invention provides an automated control system suitable for small-scale batch cultivation of highly pathogenic microorganisms. The system selects a limited set of key parameters for miniaturized microbial cultivation, providing a control system that enables coordinated manual and automated processes. On one hand, the control system utilizes limited dual-source data to simultaneously monitor and comprehensively evaluate the actual operating status of the bioreactor, thereby reducing the need for manual intervention to some extent. Simultaneously, the control system can automatically correct or review the comprehensive evaluation results (such as risk status) to further improve the sensitivity and accuracy of the control system (reducing the issuance of erroneous signals).

[0057] Furthermore, by combining it with the internal circulation condensate guiding function, the interference of condensate on pressure change values ​​can be eliminated or reduced, thereby helping to improve the monitoring accuracy of pressure sensors (such as the first pressure monitoring unit). Attached Figure Description

[0058] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0059] Figure 1a This is a schematic diagram of the first structure of a disposable bioreactor in an exemplary embodiment of the present invention;

[0060] Figure 1b is a schematic diagram of the second structure of a disposable bioreactor in an exemplary embodiment of the present invention;

[0061] Figure 1c This is a schematic diagram of the internal structure of a disposable bioreactor in an exemplary embodiment of the present invention;

[0062] Figure 1dThis is a schematic diagram of the structure of the condensate guiding region in an exemplary embodiment of the present invention;

[0063] Figure 1e This is a schematic diagram of the top wall structure in another exemplary embodiment of the present invention;

[0064] Figure 2a This is a schematic diagram of the first structure of the support device in an example embodiment of the present invention;

[0065] Figure 2b This is a schematic diagram of the second structure of the support device in an example embodiment of the present invention;

[0066] Figure 3a This is a cross-sectional schematic diagram of the sensor quick-connect device in an exemplary embodiment of the present invention;

[0067] Figure 3b This is a schematic diagram of the sensor quick-connect device in an exemplary embodiment of the present invention;

[0068] Figure 4a This is a flowchart illustrating an exemplary embodiment of the automated control method of the present invention;

[0069] Figure 4b This is a flowchart illustrating an automated control method in another exemplary embodiment of the present invention;

[0070] Figure 4c This is a schematic diagram of the modules of an automated control system in an exemplary embodiment of the present invention.

[0071] Summary of attached labeling and identification:

[0072] 1 is the reaction bag, 11 is the top wall, 12 is the annular guide rail, 12a is the annular groove, 13 is the first space, and 14 is the second space; 2 is the stirring device, 21 is the stirring paddle, 22 is the stirring shaft, and 23 is the heating unit; 3 is the working status monitoring module; 4 is the first inlet; 5 is the first outlet (also called the sampling port); 6 is the second outlet, 61 is the drying device, and 62 is the filtering device; 7 is the support body, 71 is the leakage monitoring sensor, 72 is the base, 73 is the first side wall, and 74 is the second side wall; 81 is the disposable sensor probe, 82 is the probe terminal, 83 is the power terminal, and 84 is the electrode; L1 is the horizontal plane, and L2 is the guide path. Detailed Implementation

[0073] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0074] In this document, suffixes such as "module," "part," or "unit" used to denote elements are used only for the purpose of illustrative purposes and have no specific meaning in themselves. Therefore, "module," "part," or "unit" may be used interchangeably.

[0075] In this document, the terms "upper," "lower," "inner," "outer," "front," "rear," "one end," and "the other end," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the present invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0076] In this document, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," "connected," etc., should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, a direct connection, or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0077] In this document, "and / or" includes any and all combinations of one or more of the listed related items.

[0078] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0079] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0080] As used in this specification, the term "about" typically means + / - 5% of the value, more typically + / - 4%, more typically + / - 3%, more typically + / - 2%, even more typically + / - 1%, even more typically + / - 0.5% of the value.

[0081] In this specification, certain embodiments may be disclosed in a format that falls within a certain scope. It should be understood that this...

[0082] The description "within a certain range" is merely for convenience and brevity and should not be interpreted as a rigid limitation on the disclosed range. Therefore, the description of the range should be considered as having specifically disclosed all possible subranges and the independent numeric values ​​within those ranges. For example, the range... The description should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual numbers within this range, such as 1, 2, 3, 4, 5, and 6. The above rules apply regardless of the breadth of the range.

[0083] In this article, "communication connection" can refer to a direct communication connection between two or more parties, or it can be established through a third party (e.g.,

[0084] Indirect communication connection implemented by the central control module or other central control system.

[0085] Example 1

[0086] like Figures 1a-1e As shown, in a first aspect, the present invention provides a disposable biosafety bioreactor suitable for large-scale, miniaturized microbial or cell culture in a laboratory.

[0087] The bioreactor includes a reaction bag 1, the internal containment chambers of which sequentially include a first space 13 and a second space 14; wherein, when the bioreactor is in operation, the first space 13 is used to contain gases (e.g., air, oxygen, carbon dioxide, etc.), the second space 14 is used to contain liquids (e.g., cell culture medium, water, etc.), and a condensate guiding zone is formed in the top region of the first space to guide condensate to condense in the condensate guiding zone and flow back to the second space through the guiding path provided by the condensate guiding zone;

[0088] A working status monitoring module 3 is used to monitor the working status parameters of the bioreactor. These parameters include: hydraulic pressure, foam generation, dissolved oxygen rate, CO2 concentration, pH value, and turbidity of the liquid. Accordingly, the working status monitoring module includes monitoring units: a first pressure monitoring unit for monitoring the hydraulic pressure, a foam monitoring unit for monitoring foam generation, a dissolved oxygen rate monitoring unit for monitoring the dissolved oxygen rate, a carbon dioxide monitoring unit for monitoring the CO2 concentration, a pH monitoring unit for monitoring the pH value, and a turbidity monitoring unit for monitoring turbidity. The monitoring ends (e.g., the monitoring ends of sensors) of each monitoring unit are disposed on the wall of the second space to ensure accurate monitoring of each working status parameter of the liquid.

[0089] The first space is provided with at least one first inlet 4 for introducing a first gas (e.g., compressed air, carbon dioxide, nitrogen, oxygen, etc.) and at least one second inlet for introducing a first liquid (e.g., cell culture medium, bacteria to be cultured, or water, etc.). Both the first and second inlets are located outside the condensate guiding area to prevent the first or second inlet from being blocked or contaminated by condensate during the reaction process. When the bioreactor is in operation (e.g., ...), Figure 1a (As shown in the placement diagram), the opening of the first inlet is higher than that of the second inlet.

[0090] To prevent water or other liquid substances discharged through the second inlet from flowing through the first inlet and causing blockage or contamination;

[0091] The lower region of the second space is also provided with at least one first outlet 5 for removing the liquid.

[0092] In other embodiments, taking into account the specific growth characteristics of microorganisms, some or all of the first inlet or the second inlet may be located on the wall of the second space 14, so that some gas or liquid samples can be directly introduced into the culture medium in the second space.

[0093] In some embodiments, the reaction bag is a disposable biological reaction bag, and the disposable biological reaction bag includes, from the inside out: a contact layer for direct contact with microorganisms or cell cultures, a barrier layer for isolating gases, and a structural layer for providing mechanical stability.

[0094] Specifically, in some embodiments, the volume of the reaction bag is preferably set to 1.5L, 2L, 2.5L or 5L.

[0095] Specifically, in some embodiments, the bioreactor bag is a disposable bag composed of three layers of plastic material: a first layer made of polyethylene terephthalate or LDPE to provide mechanical stability; a middle layer made of PVA or PVC to act as a gas barrier; and finally, a contact layer that comes into contact with the cell culture made of PVA or PP.

[0096] To ensure stable microbial or cell culture within a small-capacity bioreactor, it is essential to guarantee the stable operation of other structures associated with the bioreactor (such as a first inlet, a second inlet, etc.). The volume (or capacity) of the second space is typically smaller than that of the first space. Preferably, the second space occupies approximately 20%–50% of the internal chamber volume of the bioreactor.

[0097] For example, in some embodiments, when the bioreactor bag is 5L, the second space is approximately 2L.

[0098] During the experiment, it is necessary to ensure that the total amount of liquid added does not exceed the volume of the second space.

[0099] For example, in some embodiments, a boundary line is provided between the first space and the second space to determine whether the liquid addition is excessive.

[0100] It is understood that the use of "first space" and "second space" in this invention is merely a distinction made from the perspective of spatial function between the internal chambers of the reaction bag. Therefore, the first space and the second space can actually be unobstructed, interconnected spaces, such as... Figure 1a As shown.

[0101] In some embodiments, such as Figure 1d As shown, the top wall 11 inside the condensate guiding area is configured as a slope.

[0102] Furthermore, an annular guide rail 12 is provided at the edge of the condensate guiding area, and the annular guide rail 12 provides a guiding path L2 for the condensate to condense and flow back.

[0103] For example, in some embodiments, the top of the first space (or, only the condensate guiding area) is configured as a slope with a certain angle. Specifically, the top of the first space may be configured in an inverted cone shape.

[0104] For example, in some embodiments, a circular guide rail is provided at the edge of the condensate guiding area. When the air humidity in the first space is high, water droplets can be preferentially condensed on the inclined surface (i.e., the top wall 11). The water droplets flow downward along the inclined surface and gather inside the guide rail. As the gathered water droplets gradually increase in size, they flow back downward into the second space under the action of gravity.

[0105] In some embodiments, the wall of the reaction bag inside the condensate guiding zone is configured as a slope, and the inclination angle α (or slope) of the slope is approximately 1°-5°. The inclination angle refers to the angle between the wall of the condensate guiding zone and the horizontal plane L1 when the bioreactor is placed vertically.

[0106] Preferably, the inclination angle of the inclined plane is 2-3°.

[0107] Furthermore, in some embodiments, such as Figure 1e As shown, the annular guide rail is designed with a gradually decreasing curvature from its first end to its second end (i.e., the condensate outlet end). Specifically, the curvature of each point on the guide path of the annular guide rail 12 gradually decreases along the direction from its first end to its second end, so that the second end of the guide path is lower than the first end, that is, the condensate outlet end is at the lowest position.

[0108] In some embodiments, such as Figure 1c As shown, the bioreactor also includes: a stirring device 2, which includes a stirring shaft 22 and a stirring paddle 21 disposed at the end of the stirring shaft; the central area of ​​the condensate guiding zone is provided with a mounting hole for installing the stirring device 2, and the stirring shaft 22 is fixedly installed on the reaction bag through the mounting hole, while the stirring paddle 21 extends into the second space;

[0109] The stirring shaft 22 is provided with a heating unit 23 (for example, a constant temperature heating element can be provided) on the side located or close to the condensate guiding area to avoid the formation of condensate on the outside of the condensate guiding area; the monitoring end of the working status monitoring module is higher than the horizontal height of the stirring paddle to reduce the interference of the stirring paddle on the working stability of the working status monitoring module.

[0110] In this embodiment of the invention, to avoid the generation of excessive condensate in the first space due to excessively low temperature and excessively high humidity,

[0111] As a result, condensate may condense outside the condensate guiding zone or overflow from inside the condensate guiding zone. An electric heating element is installed on the stirring shaft 22 near the condensate guiding zone and away from the liquid surface. For example, the electric heating element can activate the heating function when the humidity is too high or the temperature is too low.

[0112] Understandably, since stirring devices are usually made of metal, when the heating function of the stirring device is not turned on,

[0113] A certain amount of condensate usually condenses at the top of the stirring shaft. Part of this condensate will flow back to the second space along the stirring shaft, while the other part can flow back to the second space along the guiding path under the action of the condensate guiding zone.

[0114] In some embodiments, the stirring device may be a magnetic stirring device.

[0115] In this embodiment of the invention, in order to control the volume and application cost of the miniaturized bioreactor, an internal circulation condensate recirculation scheme (without external drainage pipes) is provided. This scheme can not only avoid condensate from clogging or even contaminating the various inlets or outlets during the reaction process, but also further ensure the internal stability of the bioreactor (reducing liquid loss).

[0116] In some embodiments, such as Figure 1d As shown, the annular guide rail 12 is provided with an annular groove 12a for providing the guide path L2, and a condensate outlet is provided on the annular groove 12a. The bottom surface of the annular groove is inclined so that the condensate outlet is the lowest horizontal point of the annular groove, thereby guiding the condensate condensed in the annular groove to flow out through the condensate outlet; wherein, the condensate outlet is arranged in a direction away from the first inlet and the second inlet.

[0117] In this embodiment of the invention, the low-outlet design of the inclined groove can further guide and promote the formation of condensate within a defined area (i.e., the condensate guiding area), and prevent condensate from overflowing due to excessive condensation. Especially when culturing highly pathogenic viruses or bacteria, the concentrated formation and reflux of condensate can ensure the stable progress of the experiment, while reducing the possibility of highly pathogenic viruses or bacteria overflowing through the pipes and contaminating the external environment.

[0118] In some embodiments, the first inlet (i.e., gas inlet) for introducing gas is provided with a first control valve (i.e., air inlet valve), the second inlet (i.e., liquid inlet) for introducing liquid is provided with a second control valve (i.e., liquid inlet valve), and a second pressure monitoring unit (i.e., air pressure monitoring unit) for monitoring the air pressure inside the first space is provided in the first space, and the first pressure monitoring unit (i.e., hydraulic monitoring unit) is communicatively connected to the second control valve, and the second pressure monitoring unit is communicatively connected to both the first control valve and the second control valve.

[0119] The following describes part of the workflow of the air pressure monitoring unit, hydraulic monitoring unit, air inlet valve, and liquid inlet valve:

[0120] When the hydraulic monitoring unit detects that the hydraulic pressure exceeds a preset first pressure threshold, it sends a first shut-off signal to the inlet valve to indicate that the inlet should be stopped. The inlet valve responds to the first shut-off signal by keeping the liquid inlet closed.

[0121] When the gas pressure monitoring unit detects that the gas pressure exceeds a preset second pressure threshold, it sends a second shut-off signal to the gas inlet valve and the liquid inlet valve to indicate that the sample injection should be stopped. The gas inlet valve and the liquid inlet valve then keep the gas and liquid inlets closed based on the second shut-off signal.

[0122] In some embodiments, when hydraulic or pneumatic pressure is detected to exceed a preset pressure threshold, a warning signal can be sent directly to the user, allowing the user to intervene and determine whether to close or open each inlet.

[0123] To ensure the safety and reliability of miniaturized experiments (e.g., when the capacity of the reaction bag is only 0.5L, 1L or 2L), the embodiments of the present invention simultaneously monitor the air pressure and hydraulic pressure to avoid excessive liquid or gas addition that could lead to experimental failure or even reaction bag rupture.

[0124] Furthermore, in some embodiments, it also includes:

[0125] Corresponding to the first leakage detection module (e.g., the first leakage detection module can be a leakage sensor) disposed at the bottom of the reaction bag for monitoring whether the reaction bag leaks.

[0126] Furthermore, in some embodiments, it also includes:

[0127] The safety disinfection module includes: a recovery unit connected to the reaction bag and a disinfection unit; the recovery unit includes: a recovery bag and a recovery pipe for connecting the recovery bag and the reaction bag, the recovery pipe being provided with a third control valve (i.e., a recovery valve); the disinfection unit includes: a chamber for storing disinfectant and a disinfection pipe for connecting the chamber and the reaction bag, the disinfection pipe being provided with a fourth control valve (i.e., a disinfection valve);

[0128] The third control valve and the fourth control valve are both communicatively connected to the pressure monitoring sensor and the first leakage monitoring module.

[0129] In this embodiment of the invention, in order to control the amount of disinfectant used and reduce the impact of the disinfection process on the environment surrounding the reaction bag (e.g.,

[0130] To mitigate the adverse effects of the support device used to support the reaction bag (or the biosafety cabinet where the reaction bag is located), a process of first recycling and then sterilizing is adopted.

[0131] The following explains the preferred disinfection process for the safety disinfection module:

[0132] Step 21: The safety disinfection module (e.g., recovery valve, disinfection valve) receives an opening signal (e.g., when leakage of the reaction bag is detected, such as when the risk status is detected as leakage, or when the user inputs a disinfection command).

[0133] Step 22: The recovery valve opens and draws the liquid in the second space into the recovery bag;

[0134] After the liquid has been completely recovered, or after a certain preset recovery time (e.g., 10 seconds), proceed to step 23:

[0135] The recovery valve is closed and the disinfection valve is opened, allowing the disinfectant in the chamber to be introduced into the reaction bag.

[0136] In some embodiments, the recycling bag contains a disinfectant for disinfecting the liquid fluid.

[0137] In some embodiments, the disinfectant is a second liquid or a second gas used for disinfection, such as hydrogen peroxide gas.

[0138] In some embodiments, the recycling bag may employ a multi-layer design similar to that of a disposable bioreactor bag.

[0139] In some embodiments, after the entire bioreactor operation is completed, the safety disinfection module can disinfect the bioreactor in the same or similar manner as steps 21-23.

[0140] In this embodiment of the invention, to prevent further leakage of the liquid (containing highly hazardous microorganisms) inside the reaction bag during the filling of the disinfectant, which could even worsen the damage to the reaction bag, the liquid is recovered before filling with the disinfectant, and then disinfectant gas or liquid is introduced to centrally disinfect the reaction bag.

[0141] In some embodiments, a disinfectant substance may be introduced into the reaction bag through the first outlet.

[0142] In some embodiments, the device further includes a safety sampling device, which includes a sampling tube connected to the first outlet 5, a fifth control valve disposed inside the sampling tube, a plurality of sampling ports disposed along the axial direction on the sampling tube, and a disposable sampling bag sealed to the sampling end of the sampling port, wherein the open end of the disposable sampling bag is made of thermoplastic.

[0143] After the sampling bag has been used to collect samples, the open end of the sampling bag can be heat-treated by a clamping heater, so that the sampling bag is automatically sealed under the heat treatment and can be removed from the sampling end of the sampling port by the user.

[0144] In some embodiments, the reaction bag is connected to four gas inlets. Of course, the number and location of the gas inlets can be adjusted according to actual needs.

[0145] In some embodiments, the control valve in the gas inlet is a backflow preventer.

[0146] In some embodiments, a second outlet (i.e., an air vent) is also provided on the wall of the first space. The air vent is connected to an air vent pipe, and a filter device (e.g., an H14 HEPA filter) is also connected inside the air vent pipe to filter the aerosol inside the reaction bag and prevent pollution to the external environment. When the reaction bag is used in a bioreactor, the air vent pipe can also be connected to the exhaust pipe of the bioreactor.

[0147] In some embodiments, a drying device 61 is provided at the gas outlet. For example, a desiccant is provided inside the pipe at the gas outlet to filter contaminants in the gas.

[0148] Alternatively, in some other embodiments, when an anhydrous disinfectant is used inside the filter device 62, the desiccant can be omitted.

[0149] Furthermore, in some embodiments, by monitoring key parameters of the liquid inside the reaction bag, such as pressure (which can be used to measure the content of the reaction liquid), pH, dissolved oxygen rate, CO2 concentration, turbidity, etc., it is determined whether it is necessary to replenish cell culture medium, water, acid or alkali solution, etc., into the reactor to complete the automatic replenishment of liquid during the culture process.

[0150] Furthermore, in some embodiments, any one of the control valves can be a check valve.

[0151] Furthermore, in some embodiments, the control valve used to control the liquid can control the amount or rate of liquid addition via a peristaltic pump.

[0152] Furthermore, in some embodiments, a sterilization unit, such as a 0.22-micron sterilization filter, is provided at the inlet in the reaction bag for ventilation (e.g., oxygen).

[0153] It is understood that the disposable bioreactor in this embodiment of the invention can also be used to achieve continuous culture of microorganisms or cells. For example, taking virus culture as an example, when the number of viruses in the disposable bioreactor reaches a certain amount, a portion of the virus sample can be taken out from inside the reactor, and then new culture medium can be added to the reactor so that the remaining viruses can continue to grow inside the reactor, thereby achieving continuous virus culture.

[0154] In some embodiments, such as Figures 3a-3bAs shown, the sensor employs a biosafety-compliant quick-connect device to achieve sealing. This device includes a disposable sensor probe 81, a probe terminal 82, a power terminal 83, and electrodes 84. The probe terminal 82 contains positive and negative electrodes, which can be connected to the power terminal 83 via electrodes 84 to transmit current signals (specifically, sensor signals such as hydraulic pressure or dissolved oxygen levels) to the central control system (such as a central control module) for real-time monitoring of sensor parameters. In use, the probe terminal 82 is connected to the power terminal 83 to transmit electrical signals.

[0155] In some embodiments, the support device is provided with an interface for installing a power terminal 83. The interface of the support device is made of a corrosion-resistant sealing material. The power terminal 83 can be inserted into the interface by means of quick-connect or threaded connection to ensure the seal at the junction of the power terminal 83 and the support device and prevent aerosol leakage.

[0156] In some embodiments, for single-use bioreactors of 20L or less, the bioreactor is placed in a biosafety cabinet / isolation unit, and sampling and testing are performed directly in the biosafety cabinet / isolation unit by controlling the sampling check valve.

[0157] In some embodiments, for reactors larger than 20L, the sampling pipeline is directly connected to the biosafety cabinet / isolator, and sampling and testing are performed by controlling the sampling check valve.

[0158] Alternatively, in some embodiments, the bioreactor further includes a central control module, which is communicatively connected to one or more control valves, one or more monitoring units or monitoring modules; in order to enable real-time and accurate monitoring and maintenance of reaction safety, the central control module includes a risk monitoring unit.

[0159] Specifically, the risk monitoring unit is configured to execute the following process:

[0160] Step 11: Acquire the monitoring values ​​of the first leakage monitoring module in real time (such as the electrical signal of the leakage sensor);

[0161] Step 12: The risk monitoring unit determines the leakage risk based on the leakage value;

[0162] In order to accurately monitor the leakage situation and reduce the risk of misjudgment, the leakage risk is divided into level 0, level 1 and level 2 according to the magnitude of the leakage value.

[0163] Next, safety warnings or disinfection measures will be taken for the bioreactors according to the specific risk level of leakage.

[0164] Step 13: When the leakage risk is 1, the risk monitoring unit will determine whether the hydraulic pressure is abnormal;

[0165] If so, a corresponding disinfection signal will be sent to the recovery valve and the disinfection valve;

[0166] If not, continue to monitor the leakage risk and the hydraulic pressure. If the hydraulic pressure monitoring value increases or the hydraulic pressure becomes abnormal, send corresponding disinfection signals to the recovery valve and the disinfection valve.

[0167] Step 14: When the leakage risk is 2, send the corresponding start signal directly to the recovery valve and the disinfection valve.

[0168] In some embodiments, when a risk of leakage is detected, an early warning signal can be sent to staff, who can then manually decide whether to continue the experiment.

[0169] This invention provides a highly sensitive and relatively low-cost monitoring method (i.e., a dual-mechanism risk assessment method) by selecting two limited parameters, external leakage and internal hydraulic pressure, for monitoring.

[0170] In fact, the cultivation of highly infectious and pathogenic bacteria or viruses currently requires high-level biosafety laboratories (e.g., P3 laboratories). However, this invention integrates key status monitoring, dual-mechanism risk assessment, and safety protection functions within a single small bioreactor. This allows the small bioreactor to maintain internal environmental stability during the cultivation of highly hazardous microorganisms through monitoring key parameters and an internal circulating condensate recirculation design. Simultaneously, it enables autonomous risk monitoring and disinfection based on low-cost dual-mechanism monitoring.

[0171] Therefore, this small bioreactor can be directly applied to the cultivation of cells or microorganisms in a biosafety cabinet (BSC), which can reduce the human and material costs of cultivating highly hazardous microorganisms to a certain extent.

[0172] Example 2

[0173] In connection with the bioreactor in any embodiment of the present invention, the present invention also provides a monitoring method for using a bioreactor to achieve microbial culture.

[0174] Preferably, in some embodiments, such as Figure 4a As shown, the method includes the following steps:

[0175] S102 introduces a preset amount of gas or liquid into the reaction bag through the first or second inlet on the reaction bag to culture microorganisms or cells;

[0176] S104 monitors the hydraulic pressure of the first pressure monitoring unit and the leakage value of the first leakage monitoring module in real time.

[0177] S106 determines the risk status of the bioreactor based on the hydraulic pressure and the leakage value, the risk status including: safe status, and / or pending status, and / or leakage status; wherein, S106 includes:

[0178] S61 determines the pressure drop risk of the reaction bag based on the change parameters of the hydraulic pressure (e.g., the pressure drop value, or the pressure drop rate); wherein, when the change parameter is within a preset first pressure drop threshold, the pressure drop risk is level one; when the change parameter is within a preset second pressure drop threshold, the pressure drop risk is level two; otherwise, the pressure drop risk is level zero.

[0179] S62 determines the leakage risk of the reaction bag based on the leakage value (for example, in some embodiments, the leakage value is the electrical signal value of the leakage sensor), wherein when the leakage value is within a preset first leakage threshold, the leakage risk is level one; when the leakage value is within a preset second leakage threshold, the leakage risk is level two; otherwise, the leakage risk is level zero.

[0180] S63 determines the risk status based on the pressure reduction risk in S61 and / or the leakage risk in S62; wherein, when the pressure reduction risk is level 2 and / or the leakage risk is level 2, the risk status is a leakage status; when both the pressure reduction risk and the leakage risk are level 0, the risk status is a safe status; otherwise, the risk status is a pending status.

[0181] In some embodiments, a condensate guiding zone is further formed in the top region of the first space to guide condensate to condense within the condensate guiding zone and flow back to the second space through the guiding path provided by the condensate guiding zone. The method further includes the step of:

[0182] S108 When the risk status is detected to be either pending or leakage status, a corresponding warning signal is issued to the user respectively;

[0183] S110 responds to the feedback signal sent by the user to perform secondary monitoring of the pressure reduction risk and / or leakage risk of the bioreactor, or to receive or correct the warning signal.

[0184] In some embodiments, step S110 includes:

[0185] When the pressure drop risk is level one and the leakage risk is level zero, the condensate guide area is heated in response to the first feedback signal issued by the user.

[0186] When the pressure drop risk is detected to drop to zero during or after heating, the risk state is corrected to a safe state in response to a first correction signal issued by the user; otherwise, the risk state is corrected to a leakage state in response to a second correction signal issued by the user.

[0187] Specifically, in some embodiments, when a risk status is detected as pending, an early warning signal needs to be sent to the user to remind the user to check the working status of the bioreactor in a timely manner (e.g., whether there is a tendency to leak or a slight leakage, or whether there is excessive condensation, etc.).

[0188] For example, in some embodiments, when the risk status is detected as pending, the user will receive a corresponding warning signal. At this time, the user can choose to heat the condensate guide zone (preferably by heating through a heating unit set on the stirring shaft) to limit or reduce the formation of condensate, thereby determining whether there is a problem of excessive condensate accumulation in the current bioreactor. If so, the user can correct the risk status to a safe status when the pressure risk returns to a normal state (such as level zero).

[0189] It is understandable that, especially for bioreactors with a volume of only a few liters (e.g., 1L), the total amount of liquid inside the reactor is relatively limited, making hydraulic pressure relatively susceptible to multiple factors (such as stirring speed, stirring intensity, leakage, condensation, etc.). In this case, setting the sensor's monitoring difference (equivalent to the second pressure drop threshold) too high will make it difficult to detect problems immediately, while setting it too low may lead to frequent false alarms interfering with the normal operation of the experiment. In this embodiment of the invention, the formation or backflow of condensate is controlled or influenced by an internally circulating condensate guiding zone, so that even when the sensor is at a low monitoring difference (i.e., high sensitivity), excessive false alarms will not interfere with the normal operation of the experiment. In other words, this invention can improve the accuracy of leakage detection to a certain extent.

[0190] For example, in some embodiments, the monitoring difference can be set to around 5% or even lower. Specifically, when the difference in hydraulic pressure reaches or exceeds 5%, the bioreactor is considered to have a high risk of leakage.

[0191] Specifically, in some embodiments, when it is detected that the hydraulic pressure value recovers to the expected level (the expected level can be determined based on the actual amount of liquid added) within a preset second time period, and the risk of leakage remains unchanged, the risk state can be transformed into a safe state.

[0192] Alternatively, in some embodiments, if it is detected that the hydraulic pressure parameters do not change significantly within a preset third time period (e.g., still within the range of the first pressure drop threshold), a corresponding warning signal is issued to the user to alert the user that there is a risk of excessive condensate.

[0193] It is understood that each threshold in the embodiments of the present invention can be adaptively set by the user in combination with the actual situation (such as the volume of the reactor, the amount of liquid added, etc.).

[0194] In this embodiment of the invention, by real-time monitoring of a limited number of parameters, a low-cost and high-precision monitoring mode is provided for the working status (specifically, the risk status) of the bioreactor. On the one hand, this can reduce the manual monitoring pressure on staff to a certain extent, and on the other hand, it can avoid interference caused by over-monitoring (causing the experiment to be stopped due to error signals, resulting in a waste of human and material resources).

[0195] In some embodiments, prior to S102, the step further includes: S100 pre-testing the bioreactor;

[0196] Wherein, S100 includes:

[0197] Gas is introduced into the reaction bag through the first inlet, while the other inlets and outlets of the reaction bag are kept open.

[0198] All mouths are closed;

[0199] End ventilation and keep the corresponding first inlet closed;

[0200] The first pressure monitoring unit collects the first pressure change value of the reaction bag within a first time period, and determines whether the bioreactor passes the pre-test (i.e., whether the sealing performance is good) based on the first pressure change value; wherein, when the first pressure change value is within a preset safety threshold (specifically, when the pressure change is small or the change value is 0), the bioreactor passes the pre-test.

[0201] Example 3

[0202] like Figure 2a , 2b As shown, in order to further improve the operational safety in the process of culturing highly pathogenic microorganisms, the present invention also provides a support device for supporting the above-mentioned bioreactor.

[0203] like Figure 2a As shown, it includes:

[0204] A temperature-controlled support body 7 is used to support a bioreactor; the bioreactor includes a reaction bag, and the support body 7 includes a sidewall structure formed by a first sidewall 73 and a second sidewall 74, a base 72 formed below the sidewall structure, and a top cover for sealing the top of the sidewall structure, wherein the sidewall structure and the base 72 provide support for the reaction bag; the top cover is provided with at least one third inlet for leading out at least one pipe connected from the first inlet and the second inlet in the reaction bag, and the first and second inlets are respectively used to add gas and liquid to the reaction bag;

[0205] The heating module includes: a heating cavity disposed on the first sidewall and / or the second sidewall, the heating cavity being used to contain a heat-conducting medium, and the heating cavity also having a medium inlet and a medium outlet for the heat-conducting medium to enter and exit; a temperature monitoring unit (e.g., a temperature sensor) for monitoring the temperature of the heat-conducting medium; and a heating unit communicatively connected to the temperature monitoring unit for heating the heat-conducting medium.

[0206] And a first leakage monitoring module (such as leakage monitoring sensor 71) disposed in the central region of the bottom, the first leakage monitoring module being used to monitor whether leakage occurs in the reaction bag and / or the heating chamber.

[0207] In some embodiments, for small bioreactor bags, water is used as a heat-conducting medium to uniformly heat the circumferential walls of the small bioreactor bag.

[0208] In some embodiments, when the temperature sensor detects that the temperature inside the reactor is lower than a preset culture temperature,

[0209] A suitable high-temperature heat transfer medium can be introduced into the heating chamber to heat the reactor. Alternatively, the heat transfer medium can be directly heated by a heating unit (e.g., an electric heating element inserted into water).

[0210] In some embodiments, the bioreactor includes: a working status monitoring module for monitoring the working status parameters of the bioreactor, the working status parameters including: liquid hydraulic pressure, foam generation, dissolved oxygen rate, CO2 concentration, pH value, and turbidity; correspondingly, the support device further includes: a central control module, the central control module being communicatively connected to the working status monitoring module, the first leakage monitoring module, and the heating module, and controlling the operating status of the bioreactor and the support device according to the monitoring results communicated by the working status monitoring module, the first leakage monitoring module, and the heating module.

[0211] In some embodiments, the bioreactor further includes a safety disinfection module that is communicatively connected to the central control module;

[0212] The safety disinfection module includes a recovery unit and a disinfection unit connected to the reaction bag. The recovery unit includes a recovery bag and a recovery pipe for connecting the recovery bag and the reaction bag. A third control valve is provided in the recovery pipe. The disinfection unit includes a chamber for storing disinfectant (e.g., hydrogen peroxide disinfectant), a first disinfection pipe for introducing the disinfectant into the reaction bag, and a second disinfection pipe for introducing the disinfectant between the support body and the reaction bag. A fourth control valve is provided in both the first and second disinfection pipes. The chamber may be located on the second side wall.

[0213] Furthermore, in some embodiments, the safety disinfection module can also disinfect the various pipelines in the reaction bag or support device.

[0214] Especially when culturing highly pathogenic viruses, bacteria and other microorganisms, the support device in this embodiment of the invention can detect the working status of the disposable bioreactor in real time, and promptly activate the safety disinfection module to complete the safety disinfection when a leakage signal is detected.

[0215] In some embodiments, a viewing window is also provided on the second sidewall for observing the reaction conditions inside the reaction bag.

[0216] In some embodiments, a second leakage monitoring module is provided on the second sidewall, and when the reaction bag is installed inside the support body, the monitoring end of the second leakage monitoring module is in contact with the wall surface of the reaction bag.

[0217] Therefore, when the reaction bag is slightly damaged (such as when condensate overflows from the mounting hole of the stirring device, or when there is a small amount of leakage in the upper area of ​​the reaction bag), the leakage monitoring sensor set on the second side wall can supplement the monitoring of the stability of the reaction bag, further reducing the experimental risk.

[0218] In some embodiments, the leakage monitoring module includes one or more of the following sensors: point leakage sensor, bracket probe leakage sensor, non-positioning leakage detector, and positioning leakage detector.

[0219] In some embodiments, the top cover and the sidewall structure are threaded together.

[0220] In some embodiments, the top cover and the side wall structure are connected by a quick-connect structure, and the area where the top cover and the side wall structure contact each other is made of a corrosion-resistant material.

[0221] In some embodiments, the side wall of the support device is provided with two leak-proof disinfection ports (connected to the disinfection unit).

[0222] In the event of a leak, the cavity is circulated and disinfected through two leak-proof ports. These ports can be flexibly installed on the side wall or base of the support device.

[0223] For example, in some embodiments, one of the two leak-proof interfaces is an inlet and the other is an outlet. Specifically, during the disinfection process, a certain amount of disinfectant can be output into the support device through the inlet (e.g., liquid disinfectant / gas disinfection can be introduced or sprayed through the interface) and disinfected for a period of time, and then discharged through the outlet.

[0224] Alternatively, in some other embodiments, the input disinfectant can be recycled. Specifically, disinfectant is introduced at the inlet, and a peristaltic pump is used to recover the disinfectant at the outlet, and the recovered disinfectant is then reintroduced into the support device (e.g., by adding it back into the support device through the inlet).

[0225] In some embodiments, after disinfection is completed, the inside of the support device can be cleaned through the inlet or interface (e.g., by washing the inside with water) to extend the service life of the support device.

[0226] Understandably, staff can choose different disinfection methods based on different application needs (such as disinfection of new products or disinfection after the reaction is completed).

[0227] Example 4

[0228] The present invention also provides a monitoring method for a disposable biosafety bioreactor support device, comprising the following steps:

[0229] S200 provides any of the support devices provided by the present invention, and a reaction bag; wherein, the reaction bag comprises:

[0230] The first pressure monitoring unit for measuring hydraulic pressure, the reaction bag is installed inside the support body of the support device, the wall of the reaction bag abuts against or is adjacent to at least one side wall of the support device to improve the heating efficiency of the heating module heating the reaction bag, and the bottom of the reaction bag is in contact with the first leakage monitoring module to facilitate the first leakage monitoring module to monitor the bottom of the reaction bag.

[0231] S202 introduces a preset amount of gas or liquid into the reaction bag through the first or second inlet on the reaction bag to culture microorganisms or cells;

[0232] S204 monitors the hydraulic pressure of the first pressure monitoring unit and the leakage value of the first leakage monitoring module in real time through the central control module of the support device.

[0233] S206 determines the risk status of the support device and the bioreactor based on the hydraulic pressure and the leakage value.

[0234] In some embodiments, the risk status includes: a safe status, and / or a leakage status, and / or a pending status;

[0235] S206 includes:

[0236] The pressure reduction risk of the reaction bag is determined based on the change parameters of the hydraulic pressure; wherein, when the change parameters are at a preset first pressure reduction threshold, the pressure reduction risk is level one; when the change parameters are at a preset second pressure reduction threshold, the pressure reduction risk is level two; otherwise, the pressure reduction risk is level zero.

[0237] The leakage risk of the reaction bag is determined based on the leakage value. When the leakage value is within a preset first leakage threshold, the leakage risk is level one; when the leakage value is within a preset second leakage threshold, the leakage risk is level two; otherwise, the leakage risk is level zero.

[0238] The risk status is determined based on the pressure reduction risk and / or the leakage risk; wherein...

[0239] When the pressure drop risk is level 2 and / or the leakage risk is level 2, the risk status is a leakage status; when both the pressure drop risk and the leakage risk are level 0, the risk status is a safe status; when the pressure drop risk is level 1 and the leakage risk is level 0, or when the pressure drop risk is level 0 and the leakage risk is level 1, the risk status is a pending status.

[0240] It is understood that the monitoring method in this embodiment may include the same steps as in Embodiment 1 or 2. For example,

[0241] The method also includes: issuing warnings to users based on the results of the risk status, or disinfecting the support device and reactor, etc., which will not be elaborated here.

[0242] In some embodiments, a pressure monitoring module for monitoring the internal pressure of the support body is also provided on the second side wall of the support device. Accordingly, before S202, the step S208 is further included: S208 Pre-testing the bioreactor and the support device.

[0243] S208 includes the following steps:

[0244] Gas is introduced into the reaction bag through the first inlet, and the second gas pressure change value inside the support body is monitored by the gas pressure monitoring module; at the same time, the remaining inlets and outlets of the reaction bag and the support device are kept closed.

[0245] End ventilation and keep the corresponding first inlet closed;

[0246] The first pressure monitoring unit collects the first pressure change value of the reaction bag within a preset first time period.

[0247] The pressure monitoring module monitors the third pressure change value of the support body during the first time period.

[0248] The bioreactor is judged to pass the pre-test based on the first air pressure change value, and the support device is judged to pass the pre-test based on the second and third air pressure change values.

[0249] Specifically, when the first pressure change value is within the preset corresponding safety threshold (i.e., the pressure change is 0 or very small), the bioreactor passes the pre-test; when the second pressure change value matches the amount of gas introduced (it can be understood that the more gas introduced, the greater the pressure change value) and the third pressure change value is within the preset corresponding safety threshold, the support device passes the pre-test.

[0250] For example, in some embodiments, if the gas pressure in the reaction bag does not change significantly after ventilation, the reaction bag is considered to have good sealing performance and meet the requirements.

[0251] For example, in some embodiments, if the air pressure inside the support body increases significantly during ventilation and remains stable during the first period of testing, the support device is considered to be well-sealed and meets the requirements.

[0252] Furthermore, in some embodiments, the support device can also achieve high-temperature in-situ sterilization.

[0253] Furthermore, in some embodiments, the base is also provided with pulleys to facilitate the transportation of the support device.

[0254] Furthermore, in some embodiments, the top of the support device is also provided with a mounting hole for mounting the stirring device 2.

[0255] In some embodiments, different structures of stirring devices can be selected depending on the culture medium. For example, when the culture medium is cells, the stirring impeller of the stirring device can be blade-shaped. As another example, when the culture medium is bacteria, the stirring impeller of the stirrer can be rod-shaped.

[0256] In some embodiments, the various monitoring units equipped in the reaction bag can all be selected from disposable electrode detection sensors.

[0257] The sensors and reaction bags are all sealed to prevent leakage. The entire bioreactor requires only simple installation and is for single use.

[0258] The disposable bioreactor provided by this invention can be used for the cultivation of various microorganisms and cells. For example, the disposable bioreactor can be used in vaccine production.

[0259] It should be noted that the present invention preferably provides a small integrated bioreactor and supporting device for the cultivation of miniaturized, highly pathogenic microorganisms. Furthermore, this small integrated bioreactor can be scaled up according to the user's actual needs (e.g., from small-scale to pilot-scale), for example, scaled up to a bioreactor with a chamber capacity of approximately 100L-200L.

[0260] This invention can be widely applied to various microorganisms or cells. For example, it can be applied to the culture of highly pathogenic bacteria and viruses, as well as to other microorganisms or cells that may cause varying degrees of pollution or pathogenicity to the environment or humans. Of course, it can also be applied to the culture of various animal and plant cells.

[0261] Example 5

[0262] The present invention also provides a semi-automatic control method applicable to the above-mentioned bioreactor and support device, so as to reduce the user's intervention (such as ventilation or disinfection) to a certain extent during the culture of highly pathogenic viruses.

[0263] Figure 4b An exemplary semi-automatic control method according to the present invention is shown, comprising:

[0264] S301 provides a bioreactor system for microbial growth, the bioreactor system comprising: a bioreactor,

[0265] And a support device for supporting the bioreactor, wherein the bioreactor includes: a reaction bag, the reaction bag having a first space and a second space for containing gas and liquid respectively, the reaction bag also having a first inlet and a second inlet for introducing gas and liquid respectively, and the reaction bag also having a working status monitoring module for monitoring the working status parameters of the bioreactor;

[0266] S302 responds to a corresponding control signal issued by the user or the working status monitoring module through the first input and / or

[0267] Alternatively, a second inlet may be used to introduce gas and / or liquid into the reaction bag for biological culture (microbial culture or cell culture);

[0268] S303 performs pressure monitoring on the liquid inside the second space to collect hydraulic pressure, and performs leakage monitoring on the bottom of the reaction bag to collect leakage value;

[0269] S304 determines the risk status of the bioreactor system based on the hydraulic pressure and the leakage value, wherein the risk status includes: a safe status, and / or a pending status, and / or a leakage status; wherein, S304 includes:

[0270] The pressure reduction risk of the reaction bag is determined based on the changing parameters of the hydraulic pressure; wherein the pressure reduction risk is divided into three levels according to the magnitude of the changing parameters: level zero, level one, and level two.

[0271] The leakage risk of the reaction bag is determined based on the leakage value; wherein the leakage risk is divided into three levels according to the magnitude of the leakage value: level zero, level one, and level two.

[0272] The risk status is determined based on the pressure reduction risk and leakage risk; wherein, when the pressure reduction risk is level two and / or...

[0273] When the leakage risk is level 2, the risk status is leakage status; when both the pressure drop risk and the leakage risk are level 0, the risk status is safe status; when the pressure drop risk is level 1 and the leakage risk is level 0, or when the pressure drop risk is level 0 and the leakage risk is level 1, the risk status is pending status.

[0274] In some embodiments, the method further includes:

[0275] When S305 detects that the risk status is either pending or leaking, it will issue a corresponding warning signal to the user.

[0276] S306 responds to the feedback signal sent by the user by performing secondary monitoring of the pressure reduction risk and / or leakage risk of the bioreactor system, so as to receive or correct the warning signal.

[0277] Preferably, in some embodiments, the feedback signal can be manually issued by a staff member.

[0278] Alternatively, in other embodiments, if no feedback signal from the operator is monitored or received within a set time period (e.g., 5 seconds or 15 seconds), a computer (which can communicate with the bioreactor or support device) may also be used to respond when no feedback signal is detected or received from the operator within a set time period (e.g., 5 seconds or 15 seconds).

[0279] Based on the current risk status, a feedback signal pre-set by staff is issued.

[0280] In some embodiments, a condensate guiding zone is further formed in the top region of the first space. The condensate guiding zone guides condensate to condense within the guiding zone and to flow back to the second space through the guiding path provided by the condensate guiding zone. Accordingly, the feedback signal includes: heating the condensate guiding zone.

[0281] The first feedback signal, and the second feedback signal used to receive the warning signal, S306 includes the following steps:

[0282] When the pressure drop risk is level one and the leakage risk is level zero, the heating unit located in the condensate guiding area responds to the first feedback signal to heat the condensate guiding area;

[0283] When the pressure drop risk is detected to drop to zero during or after heating, the risk state is corrected to a safe state in response to a first correction signal issued by the user; otherwise, the risk state is corrected to a leakage state in response to a second correction signal issued by the user.

[0284] Preferably, in this embodiment of the invention, the correction of the risk status needs to be done manually by staff to avoid misjudgment of the bioreactor.

[0285] Of course, in other embodiments, the risk status can also be corrected and assessed automatically.

[0286] In some embodiments, S306 further includes the step of:

[0287] When the pressure drop risk is detected to be level zero or the leakage risk is level one, the leakage value is monitored. When the leakage value is detected to remain constant within a preset fourth time period, the risk state is corrected to a safe state in response to the first correction signal; otherwise, the risk state is corrected to a leakage state in response to the second correction signal.

[0288] For example, in some embodiments, if the leakage value does not change significantly over a period of time (such as 5s or 10s), the user can rule out the possibility of leakage in the bioreactor.

[0289] In some embodiments, the bioreactor system further includes: a stirring device, the stirring device including a stirring shaft and a stirring paddle disposed at the end of the stirring shaft; a mounting hole for mounting the stirring device is provided in the central region of the condensate guiding zone, and the stirring shaft is fixedly mounted on the reaction bag through the mounting hole; a heating unit is provided on the side of the stirring shaft located at or near the condensate guiding zone to avoid or limit the formation of condensate.

[0290] In some embodiments, the bioreactor system further includes a safety disinfection module for disinfecting the bioreactor and the support device, the safety disinfection module including a recovery unit connected to the reaction bag via a recovery pipe, and a chamber for storing disinfectant, the chamber being connected to the reaction bag and the support device via disinfection pipes respectively; the method further includes the step of:

[0291] When S307 detects that the risk state is a leakage state, it sends a corresponding disinfection signal to the safety disinfection module.

[0292] S308 The safety disinfection module responds to the disinfection signal by recovering the liquid inside the reaction bag to the recovery unit, and closes the recovery pipeline after the recovery is completed;

[0293] S309 After the recycling pipeline is closed, the safety disinfection module responds to the disinfection signal by introducing the disinfectant into the reaction bag and the support device.

[0294] In some embodiments, the step prior to 301 is further included:

[0295] S300 performs a preliminary stability test on the bioreactor system; wherein, S300 includes:

[0296] Gas is introduced into the reaction bag through the first inlet, and the second gas pressure change value inside the support device is collected simultaneously; at the same time, the other inlets and outlets of the reaction bag and the support device are kept closed.

[0297] End ventilation and keep the corresponding first inlet closed;

[0298] The first pressure change value of the reaction bag within a preset first time period and the third pressure change value of the support device within the first time period are collected.

[0299] The stability of the bioreactor system is determined based on the first, second, and third pressure change values.

[0300] Specifically, when the first pressure change value is within the corresponding preset safety threshold, and the second and third pressure change values ​​are consistent with the amount of gas introduced, the bioreactor system passes the pre-test.

[0301] For small-scale cultivation of highly pathogenic microorganisms (such as influenza virus), this invention selects the working status of the bioreactor during the cultivation process as the core monitoring factor (i.e., internal hydraulic pressure and external leakage value) to provide a low-cost semi-automatic control method that can reduce human intervention.

[0302] Furthermore, this method can be combined with a condensate guide zone to further reduce the interference of other factors on risk monitoring and reduce misjudgments, thereby reducing R&D costs to a certain extent (specifically, it can avoid terminating the experiment due to misjudgments and excessive consumption of human and material resources).

[0303] Example 6

[0304] Based on the above semi-automatic control methods, such as Figure 4c As shown, the present invention also provides a semi-automatic system, comprising:

[0305] A bioreactor system 10, comprising: a bioreactor and a support device for supporting the bioreactor, wherein the bioreactor comprises: a reaction bag, the reaction bag having a first space and a second space for containing gas and liquid respectively, and the reaction bag also having a first inlet and a second inlet for introducing gas and liquid respectively, and a working status monitoring module for monitoring the working status parameters of the bioreactor;

[0306] The sample addition module 20 is configured to introduce gas and / or liquid into the reaction bag through the first inlet and / or the second inlet for biological culture in response to a corresponding control signal issued by the user or the working status monitoring module.

[0307] The leakage monitoring module 30 is configured to monitor the pressure of the liquid inside the second space to collect hydraulic pressure, and to monitor the leakage at the bottom of the reaction bag to collect leakage value.

[0308] Risk analysis module 40 is configured to determine the risk status of the bioreactor system based on the hydraulic pressure and the leakage value, wherein the risk status includes: a safe status, and / or a pending status, and / or a leakage status; wherein,

[0309] The risk analysis module includes:

[0310] Hydraulic analysis unit 40a is configured to determine the pressure drop risk of the reaction bag based on the changing parameters of the hydraulic pressure; wherein the pressure drop risk is classified into three levels according to the magnitude of the changing parameters: level zero, level one, and level two.

[0311] Leakage analysis unit 40b is configured to determine the leakage risk of the reaction bag based on the leakage value; wherein,

[0312] The leakage risk is classified into three levels based on the leakage value: Level 0, Level 1, and Level 2.

[0313] Risk analysis unit 40c is configured to determine the risk status based on the pressure drop risk and leakage risk;

[0314] Wherein, when the pressure drop risk is level 2 and / or the leakage risk is level 2, the risk status is leakage status; when both the pressure drop risk and the leakage risk are level 0, the risk status is safe status; when the pressure drop risk is level 1 and the leakage risk is level 0, or when the pressure drop risk is level 0 and the leakage risk is level 1, the risk status is pending status.

[0315] In some embodiments, the sample addition module can be customized by the user to add culture nodes according to the culture status, such as turbidity, so as to determine when to stop the culture, when to automatically add liquid, automatically adjust pH, and introduce different gases or oxygen, etc.

[0316] In some embodiments, it also includes:

[0317] The early warning sending module 50 is configured to send an early warning when the detected risk status is either pending or leaking.

[0318] Each user receives a corresponding warning signal.

[0319] The early warning correction module 60 is configured to perform secondary monitoring of the pressure reduction risk and / or leakage risk of the bioreactor system in response to feedback signals issued by the user, so as to receive or correct the early warning signals.

[0320] In some embodiments, a condensate guiding zone is further formed in the top region of the first space. The condensate guiding zone is used to guide condensate to condense within the guiding zone and to flow back to the second space through the guiding path provided by the condensate guiding zone. Accordingly, the feedback signal includes: a first feedback signal for heating the condensate guiding zone, and a second feedback signal for receiving the warning signal. The warning correction module 60 includes:

[0321] The secondary monitoring unit 60a is configured to monitor when the pressure drop risk is level one and the leakage risk is level zero.

[0322] The condensate guiding area is heated in response to the first feedback signal;

[0323] When the correction unit 60b detects that the pressure drop risk drops to zero during or after heating, it corrects the risk state to a safe state in response to a first correction signal issued by the user; otherwise, it corrects the risk state to a leakage state in response to a second correction signal issued by the user.

[0324] It should be noted that the user in this invention can be a computer connected to the semi-automatic control system or a test operator.

[0325] It should be noted that, in addition to being applicable to the cultivation of highly pathogenic microorganisms (such as bacteria and viruses), this invention can also be applied to other microorganisms that may cause varying degrees of pollution or impact on the environment, or other microorganisms that may cause varying degrees of pathogenicity or impact on humans or animals and plants. Of course, this invention can also be applied to the cultivation of various biological cells, such as animal cells, plant cells, or single-celled organisms.

[0326] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0327] Through the above description of the embodiments, those skilled in the art can clearly understand that the methods of the above embodiments can be implemented by means of software plus necessary general-purpose hardware platforms. Of course, they can also be implemented by hardware, but in many cases the former is a better implementation method. Based on this understanding, the technical solution of the present invention, or the part that contributes to the prior art, can be embodied in the form of a software product. This computer software product is stored in a storage medium (such as ROM / RAM, magnetic disk, optical disk) and includes several instructions to cause a computer terminal (which may be a mobile phone, computer, server, or network device, etc.) to execute the methods described in the various embodiments of the present invention.

[0328] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. All of these forms are within the protection scope of the present invention.

Claims

1. An automated control method for microbial growth, characterized in that, include: S301 provides a bioreactor system for microbial growth, the bioreactor system including: a bioreactor, and a support device for supporting the bioreactor, wherein the bioreactor includes: a reaction bag, the reaction bag having a first space and a second space for containing gas and liquid respectively, the reaction bag also having a first inlet and a second inlet for introducing gas and liquid respectively, and a working status monitoring module for monitoring the working status parameters of the bioreactor; S302 responds to a corresponding control signal issued by the user or the working status monitoring module by introducing gas and / or liquid into the reaction bag through the first inlet and / or the second inlet for biological culture; S303 performs pressure monitoring on the liquid inside the second space to collect hydraulic pressure, and performs leakage monitoring on the bottom of the reaction bag to collect leakage value; S304 determines the risk status of the bioreactor system based on the hydraulic pressure and the leakage value. The risk status includes: safe status, pending status, and leakage status. S304 includes: The pressure reduction risk of the reaction bag is determined based on the change parameters of the hydraulic pressure; wherein, when the change parameters are at a preset first pressure reduction threshold, the pressure reduction risk is level one; when the change parameters are at a preset second pressure reduction threshold, the pressure reduction risk is level two; otherwise, the pressure reduction risk is level zero. The leakage risk of the reaction bag is determined based on the leakage value. When the leakage value is within a preset first leakage threshold, the leakage risk is level one; when the leakage value is within a preset second leakage threshold, the leakage risk is level two; otherwise, the leakage risk is level zero. The risk status is determined based on the pressure reduction risk and the leakage risk; wherein, when the pressure reduction risk is level 2 and / or the leakage risk is level 2, the risk status is a leakage status; when both the pressure reduction risk and the leakage risk are level 0, the risk status is a safe status; when the pressure reduction risk is level 1 and the leakage risk is level 0, or when the pressure reduction risk is level 0 and the leakage risk is level 1, the risk status is a pending status. When S305 detects that the risk status is either pending or leaking, it will issue a corresponding warning signal to the user. S306 responds to the feedback signal sent by the user by performing secondary monitoring of the pressure reduction risk and / or leakage risk of the bioreactor system, so as to receive or correct the warning signal.

2. The automated control method for microbial growth according to claim 1, characterized in that, The top area of ​​the first space also forms a condensate guiding zone, which guides condensate to condense within the guiding zone and to flow back to the second space through the guiding path provided by the condensate guiding zone. A heating unit is also provided within the condensate guiding zone. Correspondingly, the feedback signal includes: a first feedback signal for heating the condensate guiding zone, and a second feedback signal for receiving the warning signal. S306 includes the following steps: When the pressure drop risk is level one and the leakage risk is level zero, the heating unit responds to the first feedback signal to heat the condensate guide area; When the pressure drop risk is detected to drop to zero during or after heating, the risk state is corrected to a safe state in response to a first correction signal issued by the user; otherwise, the risk state is corrected to a leakage state in response to a second correction signal issued by the user.

3. The automated control method for microbial growth according to claim 2, characterized in that, S306 also includes the following steps: When the pressure drop risk is detected to be level zero or the leakage risk is level one, the leakage value is monitored. When the leakage value is detected to remain constant within a preset fourth time period, the risk state is corrected to a safe state in response to the first correction signal; otherwise, the risk state is corrected to a leakage state in response to the second correction signal.

4. An automated control method for microbial growth according to claim 2 or 3, characterized in that, The bioreactor system further includes: a stirring device, which includes a stirring shaft and a stirring paddle disposed at the end of the stirring shaft; a mounting hole for installing the stirring device is provided in the central area of ​​the condensate guiding zone, and the stirring shaft is fixedly installed on the reaction bag through the mounting hole; the heating unit is disposed on the side of the stirring shaft located or close to the condensate guiding zone to avoid or limit the formation of condensate. And / or, the wall of the reaction bag inside the condensate guiding zone is configured as a slope, and the inclination angle of the slope is 1°-5°.

5. The automated control method for microbial growth according to claim 1, characterized in that, The bioreactor system further includes a safety disinfection module for disinfecting the bioreactor and the support device. The safety disinfection module includes a recovery unit connected to the reaction bag via a recovery pipe, and a chamber for storing disinfectant. The chamber is connected to the reaction bag and the support device via disinfection pipes. The method further includes the following steps: When S307 detects that the risk state is a leakage state, it sends a corresponding disinfection signal to the safety disinfection module. S308 The safety disinfection module responds to the disinfection signal by recovering the liquid inside the reaction bag to the recovery unit, and closes the recovery pipeline after the recovery is completed; S309 After the recycling pipeline is closed, the safety disinfection module responds to the disinfection signal by introducing the disinfectant into the reaction bag and the support device.

6. The automated control method for microbial growth according to claim 1, characterized in that, The steps preceding 301 also include: S300 performs a preliminary stability test on the bioreactor system; wherein, S300 includes: Gas is introduced into the reaction bag through the first inlet, and the second gas pressure change value inside the support device is collected simultaneously; at the same time, the other inlets and outlets of the reaction bag and the support device are kept closed. End ventilation and keep the corresponding first inlet closed; The first pressure change value of the reaction bag within a preset first time period and the third pressure change value of the support device within the first time period are collected. The stability of the bioreactor system is determined based on the first, second, and third pressure change values. Specifically, the bioreactor system passes the pre-test when the first pressure change value falls within a preset safety threshold and the second and third pressure change values ​​match the amount of gas introduced.

7. An automated control system for microbial growth, characterized in that, include: A bioreactor system, comprising: a bioreactor, and a support device for supporting the bioreactor, wherein the bioreactor comprises: a reaction bag, the reaction bag having a first space and a second space for containing gas and liquid respectively, and the reaction bag also having a first inlet and a second inlet for introducing gas and liquid respectively, and a working status monitoring module for monitoring the working status parameters of the bioreactor; The sample addition module is configured to introduce gas and / or liquid into the reaction bag through the first inlet and / or the second inlet for biological culture in response to a corresponding control signal issued by the user or the working status monitoring module. The leakage detection module is configured to monitor the pressure of the liquid inside the second space to collect hydraulic pressure, and to monitor the leakage at the bottom of the reaction bag to collect leakage value. A risk analysis module is configured to determine the risk status of the bioreactor system based on the hydraulic pressure and the leakage value, wherein the risk status includes: a safe status, a pending status, and a leakage status; wherein the risk analysis module includes: A hydraulic analysis unit is configured to determine the pressure reduction risk of the reaction bag based on the changing parameters of the hydraulic pressure; wherein, when the changing parameters are at a preset first pressure reduction threshold, the pressure reduction risk is level one; when the changing parameters are at a preset second pressure reduction threshold, the pressure reduction risk is level two; otherwise, the pressure reduction risk is level zero. The leakage analysis unit is configured to determine the leakage risk of the reaction bag based on the leakage value, wherein when the leakage value is within a preset first leakage threshold, the leakage risk is level one; when the leakage value is within a preset second leakage threshold, the leakage risk is level two; otherwise, the leakage risk is level zero. The risk analysis unit is configured to determine the risk status based on the pressure reduction risk and the leakage risk; wherein, when the pressure reduction risk is level 2 and / or the leakage risk is level 2, the risk status is a leakage status; when both the pressure reduction risk and the leakage risk are level 0, the risk status is a safe status; when the pressure reduction risk is level 1 and the leakage risk is level 0, or when the pressure reduction risk is level 0 and the leakage risk is level 1, the risk status is a pending status. The early warning sending module is configured to send corresponding early warning signals to the user when the risk status is detected to be either pending or leaking. The early warning correction module is configured to perform secondary monitoring of the pressure reduction risk and / or leakage risk of the bioreactor system in response to feedback signals issued by the user, so as to receive or correct the early warning signals.

8. An automated control system for microbial growth according to claim 7, characterized in that, The top area of ​​the first space also forms a condensate guiding zone, which guides condensate to condense within the guiding zone and to flow back to the second space through the guiding path provided by the condensate guiding zone. Correspondingly, the feedback signal includes: a first feedback signal for heating the condensate guiding zone, and a second feedback signal for receiving the warning signal. The warning correction module includes: The first correction unit is configured to heat the condensate guide zone in response to a received first feedback signal when the pressure drop risk is level one and the leakage risk is level zero; and when it is detected that the pressure drop risk drops to level zero during or after heating, the risk state is corrected to a safe state in response to a first correction signal issued by the user; otherwise, the risk state is corrected to a leakage state in response to a second correction signal issued by the user.

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