Control system and method based on cell lysis process

Abstract

According to the control system and method based on the cell lysis process, the supply condition of a cell suspension is detected through a first flow detection element, the supply condition of a lysis solution is detected through a second flow detection element, and the flowing condition of a preliminary mixed solution formed by the cell suspension and the lysis solution is detected through a third flow detection element; therefore, the flow condition of each feed liquid in the whole cell lysis process is regulated and controlled, so that the mixing reaction among the cell suspension, the lysis solution and the neutralization solution is ensured to meet the matching requirement, and the batch consistency in the mass production process is also ensured; on the other hand, an inert replenishing liquid supply module is further arranged and used for supplying inert replenishing liquid to the cell suspension supply pipeline, and the inert replenishing liquid is used for pushing the cell suspension in the cell suspension supply pipeline to flow towards the first mixing point, so that the cell suspension is prevented from remaining in the cell suspension supply pipeline; loss and waste caused by the fact that the preliminary mixed liquid is left in the first mixing point are also avoided.

Description

Technical Field

[0001] The present invention relates to the technical field of cell lysis, and in particular to a control system and method based on a cell lysis process. Background Art

[0002] Cell lysis technology is a conventional means of obtaining internal substances of cells, which generally includes biological products to be lysed, lysis solutions and neutralization solutions, such as the alkaline lysis method to extract plasmid DNA; and the biological products to be lysed are generally animal, plant or microbial cell suspensions, and the lysis solutions and neutralization solutions are chemical solutions. When using, the cell suspension and the lysis solution must be fully mixed and reacted first, and then the neutralization solution and the preliminary mixed product must be repeatedly mixed to form a lysis mixed product. The lysis mixture usually includes cell lysates, proteins and their lysates, nucleic acids and their lysates, organelles and their lysates, other metabolites, etc., as well as a mixture formed by the reaction between the above substances and the used treatment agent, wherein the target product can be any one of the above substances or a mixture of several of them (in the current biological field, the target products obtained from animal cells tend to be antibodies, the target products obtained from plant cells tend to be flavonoid alkaloids, and the target products obtained from microbial cells, especially Escherichia coli, are plasmids). Correspondingly, the impurities are cell lysates, proteins and their lysates, nucleic acids and their lysates, organelles and their lysates, other metabolites, etc., except the target product, as well as a mixture formed by the reaction between the above substances and the used treatment agent.

[0003] At present, due to the different fluid concentrations and viscosities of the cell suspension, lysis solution, preliminary mixed product and neutralization solution, and the need to mix the materials according to the ratio during the cell lysis process, the feed flow rates of the materials may be different. At the same time, due to the reasons such as human operation errors or low intelligent automatic control of the liquid preparation system in the quantitative industry, the mixing reaction between the cell suspension and the lysis solution and the neutralization solution is insufficient or excessively stirred and mixed, and the ratio requirements are not met, resulting in a certain loss in the output and yield of the target product, and it is difficult to ensure the batch consistency of multiple batches of production.

[0004] Therefore, it is necessary to design a control system and method based on the cell lysis process to solve the problem of consistency between lysis batches with minimal loss of cell suspension in the prior art. Summary of the invention

[0005] In view of the deficiencies in the prior art, the object of the present invention is to provide a control system and method based on a cell lysis process, which solves the problems of cell suspension loss and batch consistency in the existing cell lysis process.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A control system based on a cell lysis process, comprising a cell suspension supply module, a lysis solution supply module, a neutralization solution supply module, a first mixing point and a second mixing point, wherein a mixed solution outlet pipeline is connected between the first mixing point and the second mixing point;

[0008] The cell suspension supply module comprises a cell suspension supply pipeline, the lysate supply module comprises a lysate supply pipeline, and the lysate supply pipeline is connected with the cell suspension supply pipeline at the first mixing point;

[0009] It also includes a first flow detection element for detecting the cumulative flow of the cell suspension flowing through the cell suspension supply pipeline; and / or, it also includes a second flow detection element for detecting the cumulative flow of the lysate flowing from the lysate supply pipeline into the first mixing point;

[0010] The neutralizing liquid supply module comprises a neutralizing liquid supply pipeline, and the neutralizing liquid supply pipeline is connected with the mixed liquid outlet pipeline at the second mixing point;

[0011] It also includes a third flow detection element, which is used to detect the cumulative flow of the preliminary mixed liquid flowing through the mixed liquid outlet pipeline;

[0012] It also includes an inert supplementary liquid supply module connected to the cell suspension supply pipeline. The inert supplementary liquid supply module is used to supply inert supplementary liquid to the cell suspension supply pipeline to enable the cell suspension in the cell suspension supply pipeline to flow continuously.

[0013] The control system based on the cell lysis process of the present invention detects the cumulative flow of the cell suspension through the first flow detection element and regulates the relevant process flow response, detects the cumulative flow of the lysate through the second flow detection element and regulates the relevant process flow response, and detects the cumulative flow of the preliminary mixed solution formed by the cell suspension and the lysate through the third flow detection element and regulates the relevant process flow response, thereby regulating the flow of each feed liquid in the entire cell lysis process, thereby ensuring that the mixing reaction between the cell suspension, the lysate and the neutralizing solution meets the ratio requirements, the cell suspension can be consumed as completely as possible without waste, and the batch consistency in the mass production process is also guaranteed; on the other hand, an inert supplementary liquid supply module is also provided, which is used to supply an inert supplementary liquid to the cell suspension supply pipeline. The inert supplementary liquid does not react with the cell suspension, the lysate and the neutralizing liquid, and its function is not only to promote the cell suspension in the cell suspension supply pipeline to flow to the first mixing point, avoiding the cell suspension remaining in the cell suspension supply pipeline, but also avoiding the preliminary mixed solution remaining in the first mixing point, causing loss and waste; and then promoting the preliminary mixed solution to promote mixing with the neutralizing solution.

[0014] Preferably, the control system further comprises an inert supplementary liquid time automatic control unit, which is communicatively connected to the inert supplementary liquid supply module and is used to control the supply time of the inert supplementary liquid; by automatically controlling the supply time of the inert supplementary liquid, it is ensured that the cell suspension remaining in the cell suspension supply pipeline is pushed into the first mixing point to react with the lysate, and then the preliminary mixed liquid is promoted to promote mixing with the neutralization liquid, which is conducive to reducing errors such as human operation errors or low automation of the liquid preparation system, thereby ensuring batch consistency during mass production.

[0015] Preferably, the lysis liquid supply module also includes a lysis liquid loop connected to the lysis liquid supply pipeline to form a lysis liquid self-circulation pipeline, the connection point between the lysis liquid supply pipeline and the lysis liquid loop is defined as a lysis liquid outlet point, the position of the second flow detection element on the lysis liquid supply pipeline is downstream of the lysis liquid outlet point, the distance from the first flow detection element to the first mixing point is D1, the distance from the lysis liquid outlet point to the first mixing point is D2, and the distance from the starting end of the cell suspension supply pipeline to the first mixing point is D3, satisfying that D2≤D1<D3; the lysis liquid supply pipeline is used to close the lysis liquid loop and open the lysis liquid supply pipeline when the cumulative flow of the cell suspension detected by the first flow detection element is greater than 0, so that the lysis liquid in the lysis liquid supply pipeline flows to the first mixing point.

[0016] The lysis solution loop and the lysis solution supply pipeline are connected to form a lysis solution self-circulation pipeline, thereby the lysis solution can be released in advance to fill the lysis solution self-circulation pipeline, the lysis solution outlet point is closer to the first mixing point relative to the starting end of the lysis solution supply pipeline, and the distance from the first flow detection element and the starting end of the cell suspension supply pipeline to the first mixing point is equal to or greater than the distance from the lysis solution outlet point to the first mixing point, thereby, when it is necessary to push the lysis solution to flow to the first mixing point, the lysis solution filling the lysis solution self-circulation pipeline can respond quickly to the release, and flow to the first mixing point at the same time or in advance relative to the cell suspension in the cell suspension supply pipeline, avoiding empty flow loss of the cell suspension; and the second flow detection element is arranged at the downstream of the lysis solution outlet point, which can accurately detect the cumulative flow of the lysis solution reacting with the cell suspension, to ensure that the mixing reaction between the cell suspension and the lysis solution and the neutralization solution meets the ratio requirements.

[0017] Preferably, the neutralizing liquid supply module also includes a neutralizing liquid circuit connected to the neutralizing liquid supply pipeline to form a neutralizing liquid self-circulation pipeline, the connection between the neutralizing liquid supply pipeline and the neutralizing liquid circuit is defined as a neutralizing liquid outlet point, the distance from the third flow detection element to the second mixing point is D4, the distance from the neutralizing liquid outlet point to the second mixing point is D5, and the distance from the starting end of the mixed liquid outlet pipeline to the second mixing point is D6, satisfying D5≤D4<D6; the neutralizing liquid supply pipeline is used to close the neutralizing liquid circuit and open the neutralizing liquid supply pipeline when the third flow detection element detects that the cumulative flow of the preliminary mixed liquid is greater than 0, so that the neutralizing liquid in the neutralizing liquid supply pipeline flows to the second mixing point.

[0018] The neutralization liquid loop and the neutralization liquid supply pipeline are connected to form a neutralization liquid self-circulation pipeline, thereby the neutralization liquid can be released early to fill the neutralization liquid self-circulation pipeline, the neutralization liquid outlet point is closer to the second mixing point relative to the starting end of the neutralization liquid supply pipeline, and the distance from the third flow detection element and the starting end of the mixed liquid outlet pipeline to the second mixing point is equal to or greater than the distance from the neutralization liquid outlet point to the second mixing point. Therefore, when it is necessary to push the neutralization liquid to flow to the second mixing point, the neutralization liquid that fills the neutralization liquid self-circulation pipeline can respond quickly to the release, and flow to the second mixing point at the same time or in advance relative to the preliminary mixed liquid in the mixed liquid outlet pipeline, avoiding empty flow loss of the preliminary mixed liquid and inability to fully mix with the neutralization liquid, thereby ensuring that the mixing reaction between the cell suspension and the lysate and the neutralization liquid meets the ratio requirements.

[0019] Preferably, the relationship between the first flow detection element, the lysate outlet point and the first mixing point also satisfies that D1+D2≤100mm, that is, under the premise of ensuring D2≤D1, the sum of the distance of the cell suspension from the first flow detection element to the first mixing point and the distance of the lysate from the lysate outlet point to the first mixing point is ≤100mm, ensuring that the cell suspension and the lysate flow to the first mixing point and mix as quickly as possible, thereby improving the mixing reaction efficiency; if D1+D2>100mm, and D2≤D1, the distance from the first flow detection element to the first mixing point is too long, However, the fluid concentration and viscosity of the cell suspension itself are greater than those of the lysis solution, and its flow rate in the cell suspension supply pipeline is slower than that of the lysis solution. When the first flow detection element detects the cell suspension and sends a signal to make the lysis solution flow out of the lysis solution outlet point, it may result in the cell suspension arriving at the first mixing point a long time after the lysis solution arrives at the first mixing point to mix with the lysis solution, resulting in a large amount of lysis solution waste. Therefore, controlling D1+D2≤100mm is conducive to reducing material waste, improving mixing efficiency, and making the mixing reaction of the cell suspension and the lysis solution more in line with the ratio requirements.

[0020] Preferably, the relationship between the third flow detection element, the neutralization liquid outlet point and the second mixing point also satisfies that D4+D5≤100mm, that is, under the premise of ensuring D5≤D4, the sum of the distance of the preliminary mixed liquid from the third flow detection element to the second mixing point and the distance of the neutralization liquid from the neutralization liquid outlet point to the second mixing point is ≤100mm, ensuring that the preliminary mixed liquid and the neutralization liquid flow to the second mixing point and mix as soon as possible, thereby improving the mixing reaction efficiency; if D4+D5>100mm, and D5≤D4, the distance from the third flow detection element to the second mixing point is too long, However, the fluid concentration and viscosity of the preliminary mixed liquid itself are greater than those of the neutralizing liquid, and its flow rate in the mixed liquid outlet pipeline is slower than that of the neutralizing liquid. When the third flow detection element detects the preliminary mixed liquid and sends a signal to make the neutralizing liquid flow out of the neutralizing liquid outlet point, it may result in the preliminary mixed liquid arriving at the second mixing point a long time after the neutralizing liquid arrives at the second mixing point to mix with the neutralizing liquid, resulting in a large amount of neutralizing liquid waste. Therefore, controlling D4+D5≤100mm is beneficial to reducing material waste, improving mixing efficiency, and making the mixing reaction of the preliminary mixed liquid and the neutralizing liquid more in line with the ratio requirements.

[0021] Preferably, the control system also includes a control module, and a first supply pump is provided on the cell suspension supply pipeline. The control module is communicatively connected with the first supply pump and is used to control the operation of the first supply pump, automatically control the first supply pump to start, and push the cell suspension to flow toward the first mixing point on the cell suspension supply pipeline.

[0022] Preferably, the control system also includes a control module, a second supply pump and a first valve are provided on the lysate supply pipeline, the first valve is located between the lysate outlet point and the first mixing point, a second valve is provided on the lysate circuit, the control module is respectively communicated with the second supply pump, the first valve, the second valve and the first flow detection element, for controlling the operation of the second supply pump, and for opening the second valve and closing the first valve before the cumulative flow of the cell suspension detected by the first flow detection element is greater than 0, so that the lysate flows in the lysate self-circulation pipeline, and for closing the second valve and opening the first valve after the cumulative flow of the cell suspension detected by the first flow detection element is greater than 0, so that the lysate flows out of the lysate outlet point and flows to the first mixing point.

[0023] The control system of the present invention automatically controls the supply of the lysate through a control module, preferentially opens the second supply pump and the second valve, so that the lysate fills the lysate self-circulating pipeline, and is ready to quickly flow from the lysate outlet point to the first mixing point. After the first flow detection element detects the cell suspension, that is, after the cell suspension is relatively close to the first mixing point, the second valve is closed and the first valve is opened, so that the lysate can directly flow out of the lysate outlet point and quickly flow to the first mixing point, thereby avoiding the cell suspension flowing to the first mixing point first, resulting in part of the cell suspension being unable to mix with the lysate, causing cell suspension loss.

[0024] Preferably, the control system also includes a control module, a third supply pump and a third valve are provided on the neutralization liquid supply pipeline, the third valve is located between the neutralization liquid outlet point and the second mixing point, and a fourth valve is provided on the neutralization liquid circuit. The control module is communicatively connected with the third supply pump, the third valve, the fourth valve and the third flow detection element respectively, for controlling the operation of the third supply pump, and for opening the fourth valve and closing the third valve before the cumulative flow of the preliminary mixed liquid detected by the third flow detection element is greater than 0, so that the neutralization liquid flows in the neutralization liquid self-circulation pipeline, and for closing the fourth valve and opening the third valve after the cumulative flow of the preliminary mixed liquid detected by the third flow detection element is greater than 0, so that the neutralization liquid flows out of the neutralization liquid outlet point and flows to the second mixing point.

[0025] The control system of the present invention automatically controls the supply of neutralizing liquid through the control module, preferentially opens the third supply pump and the fourth valve, so that the neutralizing liquid fills the neutralizing liquid self-circulating pipeline, and is ready to flow quickly from the neutralizing liquid outlet point to the first mixing point. Then, after the third flow detection element detects the preliminary mixed liquid, which means that the preliminary mixed liquid is relatively close to the second mixing point, the fourth valve is closed and the third valve is opened, so that the neutralizing liquid can directly flow out of the neutralizing liquid outlet point and quickly flow to the second mixing point, thereby avoiding the preliminary mixed liquid flowing to the first mixing point first, resulting in part of the preliminary mixed liquid being unable to mix with the neutralizing liquid, causing the loss of the preliminary mixed liquid.

[0026] Preferably, the inert supplementary liquid supply module is also connected to the lysis solution supply pipeline, and is used to supply the inert supplementary liquid to the lysis solution supply pipeline, so that the lysis solution in the lysis solution supply pipeline flows continuously; the role of using the inert supplementary liquid to push the lysis solution is that, on the one hand, it can reduce the material of the lysis solution and avoid the lysis solution remaining in the lysis solution supply pipeline; on the other hand, it can balance the pH value of the preliminary mixed solution and avoid excessive lysis solution flowing into the first mixing point to mix with the cell suspension and destroy the set mixing ratio.

[0027] Preferably, a fifth valve is provided between the inert supplementary liquid supply module and the cell suspension supply pipeline, and a sixth valve is provided between the inert supplementary liquid supply module and the lysis solution supply pipeline; after the fifth valve is opened, the inert supplementary liquid supply module and the cell suspension supply pipeline are connected, and after the sixth valve is opened, the inert supplementary liquid supply module and the lysis solution supply pipeline are connected, thereby realizing automatic control of the supply of the inert supplementary liquid, which is beneficial to reducing errors such as human operation errors or low automation of the liquid preparation system.

[0028] Preferably, a first mixing device is connected downstream of the first mixing point, a first mixing section is connected between the first mixing point and the inlet end of the first mixing device, and the length of the first mixing section is d1, satisfying d1≤100 mm.

[0029] The first mixing point is used for the initial meeting and mixing of the cell suspension and the lysate, the first mixing device is used for the full mixing of the cell suspension and the lysate, and the length of the first mixing section determines the distance from the first mixing point to the first mixing device. Therefore, the length of the first mixing section is controlled within 100 mm so that the initially met cell suspension and lysate can be quickly moved to the first mixing device for uniform mixing; if the length of the first mixing section is greater than 100 mm, part of the cell suspension and the lysate will mix and react in the first mixing section, and the concentration and viscosity of the feed fluid will increase, which may clog the first mixing section, resulting in the inability of the subsequent cell suspension and lysate to meet at the first mixing point at the set flow rate, affecting the timing accuracy of the entire control system.

[0030] Preferably, a second mixing device is connected downstream of the second mixing point, a second mixing section is connected between the second mixing point and the inlet end of the second mixing device, and the length of the second mixing section is d2, satisfying d2≤100 mm.

[0031] The second mixing point is used for the preliminary meeting and mixing of the preliminary mixed liquid and the neutralizing liquid, and the second mixing device is used for the sufficient mixing of the preliminary mixed liquid and the neutralizing liquid, and the length of the second mixing section determines the distance from the second mixing point to the second mixing device. Therefore, the length of the second mixing section is controlled within 100 mm so that the preliminary mixed liquid and the neutralizing liquid that initially meet can be quickly moved to the second mixing device for uniform mixing; if the length of the second mixing section is greater than 100 mm, the preliminary mixed liquid itself is very viscous, and it cannot be quickly mixed with the neutralizing liquid in the second mixing section, and then it is easy to clog the second mixing section, resulting in the subsequent preliminary mixed liquid and neutralizing liquid being unable to meet at the second mixing point according to the set flow rate, affecting the timing control accuracy of the entire control system.

[0032] Preferably, the first mixing device comprises a first spiral mixer and a coil mixer; and / or the second mixing device comprises a second spiral mixer.

[0033] The cell suspension and the lysate are fully mixed in the first spiral mixer, and then fully reacted in the coil mixer to form a preliminary mixed solution; the preliminary mixed solution and the neutralization solution are fully mixed and reacted in the second spiral mixer to form a final mixture; the final mixture is subsequently clarified and purified to obtain the target product plasmid.

[0034] Preferably, the cell suspension supply module and the lysis liquid supply module are arranged longitudinally, the cell suspension supply pipeline includes a first transverse flow segment connected to the first mixing point, the lysis liquid supply pipeline includes a second transverse flow segment and a first longitudinal flow segment, the second transverse flow segment is located below the first transverse flow segment, and the first longitudinal flow segment connects the second transverse flow segment and the first mixing point from bottom to top.

[0035] Since the cell suspension contains solids such as cells, the solid content of the cell suspension is high, the viscosity is large, and the flow rate is slow. Therefore, the cell suspension supply pipeline is basically arranged horizontally, that is, it includes a first horizontal flow section connected to the first mixing point, so that the cell suspension flows smoothly in the cell suspension supply pipeline and is less affected by gravity, which is conducive to ensuring the stability and uniformity of the cell suspension liquid performance and flow rate; the fluid concentration and viscosity of the lysate are small, and the flow rate is more controllable. Therefore, the lysate supply pipeline includes a second horizontal flow section arranged horizontally and a first longitudinal flow section arranged vertically connecting the second horizontal flow section and the first mixing point. The lysate flows smoothly in the second horizontal flow section and flows upward in the first longitudinal flow section. Since the adhesion between the lysate and the inner wall of the tube is small, it is easier to flow upward and control the flow rate.

[0036] Preferably, the mixed liquid outlet pipeline of the first mixing point and the second mixing point are located above the neutralization liquid supply module, and the neutralization liquid supply pipeline includes a third transverse flow segment and a second longitudinal flow segment, and the second longitudinal flow segment connects the third transverse flow segment and the second mixing point from bottom to top.

[0037] Since the preliminary mixed liquid contains solids such as cell fragments and biological products, the solid content and viscosity of the preliminary mixed liquid are greater than the fluid concentration and viscosity of the cell suspension. Therefore, the mixed liquid outlet pipeline of the first mixing point and the second mixing point are located above the neutralizing liquid supply module. The gravity of the preliminary mixed liquid can be used to promote the preliminary mixed liquid to flow more smoothly in the mixed liquid outlet pipeline to the second mixing point. The fluid concentration and viscosity of the neutralizing liquid are smaller, and the flow rate is more controllable. Therefore, the neutralizing liquid supply pipeline includes a third transverse flow section arranged laterally and a second longitudinal flow section arranged vertically to connect the third transverse flow section and the second mixing point. The neutralizing liquid flows smoothly in the third transverse flow section and flows upward in the second longitudinal flow section. Since the adhesion between the neutralizing liquid and the inner wall of the tube is small, it is easier to flow upward and control the flow rate.

[0038] Preferably, the first supply pump on the cell suspension supply pipeline is arranged on the first lateral flow section, and the first flow detection element is located upstream of the first supply pump.

[0039] It takes a certain amount of time for the first flow detection element to detect the cell suspension and send a signal to make the lysis solution flow out of the lysis solution outlet point. The first flow detection element is arranged upstream of the first supply pump, so that the lysis solution can receive the signal of the presence of the cell suspension earlier and be released in time to flow to the first mixing point, thereby avoiding the cell suspension flowing to the first mixing point before the lysis solution and causing the loss of the cell suspension.

[0040] Preferably, the second supply pump on the lysate supply pipeline is arranged on the second transverse flow section, and the second flow detection element is located downstream of the second supply pump and is arranged on the first longitudinal flow section.

[0041] The second supply pump is arranged on the second transverse flow section, which is beneficial to the smooth flow of the lysis solution in the lysis solution supply pipeline, and the function of the second flow detection element is to detect the cumulative flow of the lysis solution out of the outlet point. The first longitudinal flow section is a flow section closer to the first mixing point, which ensures the accuracy of the second flow detection element in detecting the cumulative flow of the lysis solution, thereby ensuring that the mixing reaction between the cell suspension, the lysis solution and the neutralization solution meets the ratio requirements, the cell suspension is consumed as completely as possible without waste, and the batch consistency in the mass production process is also guaranteed.

[0042] In order to achieve the above object, the present invention also adopts the following technical solutions:

[0043] A control method based on a cell lysis process, using the above control system, comprises the following steps:

[0044] Mixing step: opening the cell suspension supply line, opening the lysis solution supply line, and opening the neutralization solution supply line;

[0045] The lysate reaches the first mixing point before the cell suspension or reaches the first mixing point at the same time;

[0046] The lysate and the cell suspension are mixed at the first mixing point to form a preliminary mixed solution;

[0047] The neutralizing liquid reaches the second mixing point before the preliminary mixed liquid or reaches the second mixing point at the same time, and is mixed at the second mixing point;

[0048] Emptying step: When the cumulative flow of the cell suspension detected by the first flow detection element reaches a first flow setting value or the cumulative flow of the lysate detected by the second flow detection element reaches a second flow setting value, the inert supplementary liquid supply module supplies an inert supplementary liquid to the cell suspension supply pipeline to allow the cell suspension in the cell suspension supply pipeline to flow continuously until the inert supplementary liquid supply time is reached.

[0049] The control method based on the cell lysis process of the present invention controls the lysis solution to reach the first mixing point before the cell suspension or to reach the first mixing point at the same time, so as to avoid the loss of the cell suspension caused by the inability of part of the cell suspension to contact and mix with the lysis solution, and controls the neutralization solution to reach the second mixing point before the preliminary mixed solution or to reach the second mixing point at the same time, so as to avoid the loss and waste of the preliminary mixed solution caused by the inability of part of the preliminary mixed solution to contact and mix with the neutralization solution; in addition, the first flow setting value refers to the cumulative flow of the cell suspension when the cell suspension is close to being emptied, and the second flow setting value refers to the cumulative flow of the lysis solution when the lysis solution is mixed with most of the cell suspension in proportion. The first flow setting value or the second flow setting value indicates that the cell suspension is close to being emptied, and then an inert supplementary liquid supply module is used to supply the inert supplementary liquid to the cell suspension supply pipeline, so that the cell suspension in the cell suspension supply pipeline flows continuously until the cell lysis reaction is completely completed, so as to avoid the cell suspension remaining in the cell suspension supply pipeline, and also avoid the preliminary mixed solution remaining in the first mixing point, causing loss and waste.

[0050] Preferably, the time when the cumulative flow rate of the cell suspension detected by the first flow detection element is greater than 0 is defined as the first time, the time when the cumulative flow rate of the lysate detected by the second flow detection element is greater than 0 is defined as the second time, and the time when the cumulative flow rate of the preliminary mixed solution detected by the third flow detection element is greater than 0 is defined as the third time;

[0051] In the emptying step, the inert replenishing liquid supply time of the inert replenishing liquid supply module is determined according to the difference between the third time and the first time; or, the inert replenishing liquid supply time of the inert replenishing liquid supply module is determined according to the difference between the third time and the second time; or, the inert replenishing liquid supply time of the inert replenishing liquid supply module is executed according to a preset time.

[0052] The difference between the third time and the first time indicates the time taken for the first drop of cell suspension to flow from the position of the first flow detection element to the position of the third flow detection element, and the inert supplementary liquid begins to be replenished into the cell suspension supply pipeline at the time point when the cell suspension is nearly emptied, which can be considered as pushing the last drop of cell suspension to flow from the position of the first flow detection element. Therefore, the supply time of the inert supplementary liquid should continue until the time point when the last drop of cell suspension flows to the position of the third flow detection element, so that all cell suspensions can be pushed to the first mixing point for mixing reaction, thereby avoiding waste of cell suspension. Similarly, the difference between the third time and the second time indicates the time taken for the first drop of lysate to flow from the first flow detection element to the position of the third flow detection element. Since the lysate is proportioned with the cell suspension, the release of the cell suspension can be judged by the accumulated flow of the lysate, thereby ensuring that the inert supplementary liquid pushes all the cell suspension to the first mixing point for mixing reaction, thereby avoiding waste of the cell suspension. Moreover, since the flow rate of the lysate is more uniform, the difference between the third time and the second time is more accurate, and the supply time of the inert supplementary liquid is more precise. Of course, in long-term experiments, practical observations or simulations, the supply time of the inert supplementary liquid can be roughly determined. Therefore, the supply time of the inert supplementary liquid of the inert supplementary liquid supply module can also be executed according to the preset time, which is suitable for batch production.

[0053] Preferably, the lysate supply module further comprises a lysate loop connected to the lysate supply pipeline to form a lysate self-circulating pipeline, and the neutralizing liquid supply module further comprises a neutralizing liquid loop connected to the neutralizing liquid supply pipeline to form a neutralizing liquid self-circulating pipeline.

[0054] The mixing step also includes a starting step:

[0055] Open the lysate self-circulation pipeline so that the lysate fills the lysate self-circulation pipeline;

[0056] Open the neutralization liquid self-circulation pipeline so that the neutralization liquid fills the neutralization liquid self-circulation pipeline;

[0057] Opening the cell suspension supply pipeline, so that the cell suspension flows to the first flow detection element;

[0058] The mixing step further comprises:

[0059] When the accumulated flow rate of the cell suspension detected by the first flow detection element is greater than 0, closing the lysate loop, opening the lysate supply pipeline, allowing the lysate in the lysate supply pipeline to flow to the first mixing point, and the lysate is mixed with the cell suspension at the first mixing point;

[0060] When the cumulative flow of the preliminary mixed liquid detected by the third flow detection element is greater than 0, the neutralizing liquid circuit is closed, the neutralizing liquid supply pipeline is opened, and the neutralizing liquid in the neutralizing liquid supply pipeline flows to the second mixing point, and the neutralizing liquid is mixed with the preliminary mixed liquid at the second mixing point.

[0061] The control method of the present invention preferentially opens the lysate self-circulating pipeline, so that the lysate fills the lysate self-circulating pipeline, and is ready to flow quickly from the lysate outlet point to the first mixing point. Then, after the first flow detection element detects the cell suspension, that is, after the cell suspension is relatively close to the first mixing point, the lysate loop is closed, and the lysate can directly flow out of the lysate outlet point and quickly flow to the first mixing point, avoiding the cell suspension flowing to the first mixing point first, resulting in part of the cell suspension being unable to mix with the lysate, causing cell suspension loss; similarly, the control method of the present invention preferentially opens the neutralization liquid self-circulating pipeline, so that the neutralization liquid fills the neutralization liquid self-circulating pipeline, and is ready to flow quickly from the neutralization liquid outlet point to the first mixing point. Then, after the third flow detection element detects the preliminary mixed liquid, that is, after the preliminary mixed liquid is relatively close to the second mixing point, the neutralization liquid loop is closed, and the neutralization liquid can directly flow out of the neutralization liquid outlet point and quickly flow to the second mixing point, avoiding the preliminary mixed liquid flowing to the first mixing point first, resulting in part of the preliminary mixed liquid being unable to mix with the neutralization liquid, causing preliminary mixed liquid loss.

[0062] In summary, compared with the prior art, the present invention has at least the following beneficial effects:

[0063] The control system and control method based on the cell lysis process of the present invention detect the supply of the cell suspension through a first flow detection element, detect the supply of the lysate through a second flow detection element, and detect the flow of the preliminary mixed solution formed by the cell suspension and the lysate through a third flow detection element, thereby regulating the flow of each feed liquid in the entire cell lysis process, thereby ensuring that the mixing reaction between the cell suspension, the lysate and the neutralizing solution meets the ratio requirements, the cell suspension can be completely consumed without waste, and the batch consistency in the mass production process is also guaranteed; on the other hand, an inert supplementary liquid supply module is also provided, which is used to supply an inert supplementary liquid to the cell suspension supply pipeline. The inert supplementary liquid does not react with the cell suspension, the lysate and the neutralizing solution, and its function is to promote the cell suspension in the cell suspension supply pipeline to flow to the first mixing point, thereby avoiding the cell suspension remaining in the cell suspension supply pipeline, and also avoiding the preliminary mixed solution remaining in the first mixing point, causing loss and waste. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0065] Figure 1 It is a schematic diagram of the framework of the control system of the first embodiment of the present invention.

[0066] Figure 2 It is a schematic diagram of the framework of the control system of the second embodiment of the present invention.

[0067] Figure 3 This is a schematic diagram of the framework of the control system of the third embodiment of the present invention.

[0068] Figure 4 It is a schematic diagram of the framework of the control system of the fourth embodiment of the present invention.

[0069] Description of Reference Numerals

[0070] 10. Cell suspension supply pipeline; 11. First flow detection element; 12. First supply pump; 13. First transverse flow section; 14. Third longitudinal flow section; 15. Cell suspension bag;

[0071] 20. Lysis solution supply pipeline; 21. Second flow detection element; 22. Lysis solution circuit; 23. Second supply pump; 24. First valve; 25. Second valve; 26. Second transverse flow section; 27. First longitudinal flow section; 28. Lysis solution outlet point; 29. ​​Lysis solution bag;

[0072] 30. Neutralizing liquid supply pipeline; 31. Neutralizing liquid circuit; 32. Third supply pump; 33. Third valve; 34. Fourth valve; 35. Third transverse flow section; 36. Second longitudinal flow section; 37. Neutralizing liquid outlet point; 38. Neutralizing liquid bag;

[0073] 40. First mixing point; 41. Mixed liquid outlet pipeline; 42. Third flow detection element; 43. First spiral mixer; 44. Coil mixer; 45. First mixing section;

[0074] 50. Second mixing point; 51. Second spiral mixer; 52. Second mixing section;

[0075] 60. Inert supplementary liquid supply module; 61. Fifth valve; 62. Sixth valve. DETAILED DESCRIPTION

[0076] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0077] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0078] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0079] The general process of cell lysis is to fully mix the cell suspension with the lysis solution to lyse the cells in the cell suspension and release the lysis mixture in the cells. The lysis mixture usually includes cell lysates, proteins and their lysates, nucleic acids and their lysates, organelles and their lysates, other metabolites, etc., as well as a mixture formed by the reaction between the above substances and the used treatment agent, wherein the target product can be any one of the above substances or a mixture of several of them (in the current biological field, the target products usually obtained from animal cells tend to be antibodies, the target products obtained from plant cells tend to be flavonoid alkaloids, and the target products obtained from microbial cells, especially Escherichia coli, are plasmids). Correspondingly, impurities are cell lysates, proteins and their lysates, nucleic acids and their lysates, organelles and their lysates, other metabolites, etc., except the target product, as well as a mixture formed by the reaction between the above substances and the used treatment agent.

[0080] Taking the alkaline lysis plasmid extraction method of obtaining plasmids from Escherichia coli as an example, the alkaline lysis method is based on the difference in denaturation and renaturation between chromosomal DNA and plasmid DNA to achieve the purpose of separation. Therefore, this method generally includes three steps: 1. Provide a certain proportion of resuspension solution to resuspend the bacterial solution (containing Escherichia coli), and add lysis solution (currently mainly alkaline reagent solution: such as 0.2M 1. After the bacterial solution and the lysis solution are fully mixed and reacted, the lysis products produced include the target plasmid and bacterial cell lysates, proteins and their lysates, chromosomal DNA and their lysates, RNA, organelles and their lysates, other metabolites, etc., and the complex or mixture formed by the reaction between the above substances and the used treatment agent; 2. When the acidic salt solution is added for neutralization, the supercoiled plasmid DNA is renatured, and the high concentration of KAC also plays a role of salting out, so that the formed SDS-gDNA-protein insoluble complex is precipitated, and the denatured proteins and lipids are turned into insoluble curd-like precipitates, and a part of the chromosomal DNA is also precipitated, while the plasmid DNA is still in a dissolved state, and the effective separation of the target product is achieved by solid-liquid separation.

[0081] At present, in the cell lysis process, each material needs to be mixed according to a ratio. The cell suspension, lysis solution, preliminary mixed product and neutralization solution have different fluid concentrations and viscosities, which makes the feed flow rates of each material different. Therefore, the control system of the embodiment of the present invention has been improved to regulate the flow of each material in the entire cell lysis process. Figures 1 to 4 In each embodiment shown, the control system includes a cell suspension supply module, a lysate supply module, a neutralizing liquid supply module, a first mixing point 40 and a second mixing point 50, and a mixed liquid outlet pipeline 41 is connected between the first mixing point 40 and the second mixing point 50; the cell suspension supply module corresponds to a cell suspension storage device such as a cell suspension bag 15 or a cell suspension cylinder, the lysate supply module corresponds to a lysate storage device such as a lysate bag 29 or a lysate cylinder, and the neutralizing liquid supply module is connected to a neutralizing liquid storage device such as a neutralizing liquid bag 38 or a neutralizing liquid cylinder, and these storage devices all include a liquid inlet valve and a liquid outlet valve to control the flow of materials out of or in from the corresponding storage device.

[0082] The cell suspension supply module includes a cell suspension supply pipeline 10, which is connected to the liquid outlet valve of the cell suspension storage device. The lysate supply module includes a lysate supply pipeline 20, which is connected to the liquid outlet valve of the lysate storage device. The lysate supply pipeline 20 is connected to the cell suspension supply pipeline 10 at the first mixing point 40; in some embodiments, the system also includes a first flow detection element 11, which is used to detect the cumulative flow of the cell suspension flowing through the cell suspension supply pipeline 10 and regulate the relevant process flow response. In some embodiments, the system also includes a second flow detection element 21, which is used to detect the cumulative flow of the lysate flowing from the lysate supply pipeline 20 into the first mixing point 40 and regulate the relevant process flow response. Since there is a proportional correspondence between the cell suspension and the lysate, the cumulative flow of one can be inferred from the cumulative flow of the other. Therefore, in some embodiments, only the first flow detection element 11 is set, or only the second flow detection element 21 is set. As a preferred embodiment, Figure 1 In the illustrated embodiment 1, the cell suspension supply pipeline 10 is provided with a first flow detection element 11 , and the lysis solution supply pipeline 20 is provided with a second flow detection element 21 .

[0083] The neutralizing liquid supply module includes a neutralizing liquid supply pipeline 30, the neutralizing liquid supply pipeline 30 is connected to the liquid outlet valve of the neutralizing liquid storage device, the neutralizing liquid supply pipeline 30 is connected to the mixed liquid outlet pipeline 41 at the second mixing point 50, and also includes a third flow detection element 42 for detecting the cumulative flow of the preliminary mixed liquid flowing through the mixed liquid outlet pipeline 41 and regulating the relevant process flow response, specifically, as Figure 1 In the illustrated embodiment, the third flow detection element 42 is disposed on the mixed liquid outlet pipeline 41. The control system based on the cell lysis process of the present invention detects the supply of the cell suspension through the first flow detection element 11, detects the supply of the lysate through the second flow detection element 21, and detects the flow of the preliminary mixed liquid formed by the cell suspension and the lysate through the third flow detection element 42, thereby regulating the flow of each feed liquid in the entire cell lysis process, thereby ensuring that the mixing reaction between the cell suspension, the lysate and the neutralization liquid meets the ratio requirements, the cell suspension can be completely consumed without waste, and the batch consistency in the mass production process is also ensured.

[0084] The system of the embodiment of the present invention also includes an inert supplementary liquid supply module 60, which is connected to the cell suspension supply pipeline 10. The inert supplementary liquid supply module 60 is used to supply an inert supplementary liquid to the cell suspension supply pipeline 10 so that the cell suspension in the cell suspension supply pipeline 10 flows continuously. The inert supplementary liquid does not react with the cell suspension, the lysis solution and the neutralization solution. Its function is not only to promote the cell suspension in the cell suspension supply pipeline 10 to flow to the first mixing point 40, to avoid the cell suspension remaining in the cell suspension supply pipeline 10, but also to avoid the preliminary mixed liquid remaining in the first mixing point 40, causing loss and waste; and then promote the preliminary mixed liquid to promote mixing with the neutralization solution.

[0085] In order to ensure that there is no residue of the cell suspension and the preliminary mixed solution, the control system further includes an inert supplementary solution time automatic control unit (not shown), which is communicatively connected to the inert supplementary solution supply module 60 and is used to control the supply time of the inert supplementary solution; by automatically controlling the supply time of the inert supplementary solution, it is ensured that the residual cell suspension in the cell suspension supply pipeline 10 is pushed into the first mixing point 40 to react with the lysing solution, and then the preliminary mixed solution is pushed to promote mixing with the neutralizing solution.

[0086] Specifically, the control logic of the inert supplementary liquid time automatic control unit has the following three types. The time when the cumulative flow rate of the cell suspension detected by the first flow detection element 11 is greater than 0 is defined as the first time, the time when the cumulative flow rate of the lysate detected by the second flow detection element 21 is greater than 0 is defined as the second time, and the time when the cumulative flow rate of the preliminary mixed solution detected by the third flow detection element 42 is greater than 0 is defined as the third time, and the inert supplementary liquid supply time of the inert supplementary liquid supply module 60 is determined according to the difference between the third time and the first time; or, the inert supplementary liquid supply time of the inert supplementary liquid supply module 60 is determined according to the difference between the third time and the second time; or, the inert supplementary liquid supply time of the inert supplementary liquid supply module 60 is executed according to the preset time.

[0087] The difference between the third time and the first time indicates the time when the first drop of cell suspension flows from the position of the first flow detection element 11 to the position of the third flow detection element 42, and the inert supplementary liquid begins to be replenished to the cell suspension supply pipeline 10 at the time point when the cell suspension is close to being emptied, which can be considered as pushing the last drop of cell suspension to flow from the position of the first flow detection element 11. Therefore, the supply time of the inert supplementary liquid should continue until the time point when the last drop of cell suspension flows to the position of the third flow detection element 42, so that all cell suspensions can be pushed to the first mixing point 40 for mixing reaction to avoid cell suspension waste. Similarly, the difference between the third time and the second time indicates the time when the first drop of lysate flows from the first flow detection element 11 to the position of the third flow detection element 42. The time for the liquid to flow from the position of the detection element 11 to the position of the third flow detection element 42. Since the lysate is mixed with the cell suspension in proportion, the release of the cell suspension can be judged by the cumulative flow of the lysate, thereby ensuring that the inert supplementary liquid pushes all the cell suspension to the first mixing point 40 for mixing reaction, avoiding waste of the cell suspension, and since the flow rate of the lysate is more uniform, the difference between the third time and the second time is more accurate, and the supply time of the inert supplementary liquid is more precise; of course, in long-term experiments or practical observations or simulations, the supply time of the inert supplementary liquid can be roughly determined, therefore, the supply time of the inert supplementary liquid of the inert supplementary liquid supply module 60 can also be executed according to the preset time, which is suitable for batch production.

[0088] like Figure 1 In the illustrated embodiment 1, the inert supplementary liquid supply module 60 is also connected to the lysis liquid supply pipeline 20, and is used to supply the inert supplementary liquid to the lysis liquid supply pipeline 20, so that the lysis liquid in the lysis liquid supply pipeline 20 flows continuously; the role of using the inert supplementary liquid to push the lysis liquid is that, on the one hand, it can reduce the amount of lysis liquid used and avoid the lysis liquid remaining in the lysis liquid supply pipeline 20; on the other hand, it can balance the pH value of the preliminary mixed liquid and avoid excessive lysis liquid flowing into the first mixing point 40 to mix with the cell suspension and destroy the set mixing ratio.

[0089] Of course, in other embodiments, such as Figure 2 In the second embodiment shown, the inert supplementary liquid supply module 60 can be connected only to the cell suspension supply pipeline 10, and not to the lysis solution supply pipeline 20, and the lysis solution supply pipeline 20 can stop supplying the lysis solution within a set time after the inert supplementary liquid supply module 60 starts to supply the inert supplementary liquid, so as to ensure that the cumulative flow rate of the lysis solution meets the ratio requirement with the cell suspension.

[0090] Specifically, Figure 1In the illustrated embodiment 1, a fifth valve 61 is provided between the inert supplementary liquid supply module 60 and the cell suspension supply pipeline 10, and a sixth valve 62 is provided between the inert supplementary liquid supply module 60 and the lysis solution supply pipeline 20; after the fifth valve 61 is opened, the inert supplementary liquid supply module 60 and the cell suspension supply pipeline 10 are connected, and after the sixth valve 62 is opened, the inert supplementary liquid supply module 60 and the lysis solution supply pipeline 20 are connected, thereby realizing automatic control of the supply of the inert supplementary liquid, which is beneficial to reducing errors such as human operation errors or low automation of the liquid preparation system.

[0091] In some embodiments of the present invention, Figure 2 Embodiment 2 and Figure 4 In the fourth embodiment, the lysis solution supply module is a single-line flow, that is, it only includes the lysis solution supply pipeline 20 and the components thereon, and the neutralization solution supply module is a single-line flow, that is, it only includes the neutralization solution supply pipeline 30 and the components thereon. In these embodiments, the time for the lysis solution to flow to the first mixing point 40 can be controlled by controlling the release time and length of the lysis solution supply pipeline 20, so that the lysis solution arrives at the first mixing point 40 before the cell suspension or at the same time as the cell suspension, and the time for the lysis solution to flow to the second mixing point 50 can be controlled by controlling the release time and length of the neutralization solution supply pipeline 30, so that the neutralization solution arrives at the second mixing point 50 before the preliminary mixed solution or at the same time as the preliminary mixed solution.

[0092] In some preferred embodiments, Figure 1 Embodiment 1 and Figure 3In the third embodiment, the lysate supply module also includes a lysate loop 22 connected to the lysate supply pipeline 20 to form a lysate self-circulating pipeline, thereby the lysate can be released in advance to fill the lysate self-circulating pipeline; the connection between the lysate supply pipeline 20 and the lysate loop 22 is defined as a lysate outlet point 28, the lysate outlet point 28 is closer to the first mixing point 40 relative to the starting end of the lysate supply pipeline 20, the route is shorter, and the time for the lysate to reach the first mixing point 40 is more controllable, the distance from the first flow detection element 11 to the first mixing point 40 is D1, the distance from the lysate outlet point 28 to the first mixing point 40 is D2, and the starting end of the cell suspension supply pipeline 10 is D1. The distance from the starting end to the first mixing point 40 is D3, which satisfies D2≤D1<D3; the lysate supply pipeline 20 is used to close the lysate loop 22 and open the lysate supply pipeline 20 when the cumulative flow of the cell suspension detected by the first flow detection element 11 is greater than 0, so that the lysate in the lysate supply pipeline 20 flows to the first mixing point 40, and the cell suspension in the cell suspension supply pipeline 10 flows to the first mixing point 40 at the same time or in advance, thereby avoiding the loss of cell suspension cavitation, and taking into account that under different flow rate ratios, the lysate and the cell suspension basically reach the first mixing point 40 at the same time or the lysate reaches the first mixing point 40 slightly earlier than the cell suspension, thereby reducing material waste. The second flow detection element 21 is arranged at the downstream of the lysate outlet 28, and can accurately detect the cumulative flow of the lysate reacting with the cell suspension, and ensure that the mixing reaction between the cell suspension, the lysate and the neutralizing solution meets the ratio requirements.

[0093] like Figure 1 Embodiment 1 and Figure 3In the third embodiment, the neutralizing liquid supply module also includes a neutralizing liquid loop 31 connected to the neutralizing liquid supply pipeline 30 to form a neutralizing liquid self-circulating pipeline, thereby the neutralizing liquid can be released early to fill the neutralizing liquid self-circulating pipeline; the connection between the neutralizing liquid supply pipeline 30 and the neutralizing liquid loop 31 is defined as a neutralizing liquid outlet point 37, and the neutralizing liquid outlet point 37 is closer to the second mixing point 50 relative to the starting end of the neutralizing liquid supply pipeline 30, the route is shorter, and the time for the neutralizing liquid to reach the second mixing point 50 is more controllable; the distance from the third flow detection element 42 to the second mixing point 50 is D4, the distance from the neutralizing liquid outlet point 37 to the second mixing point 50 is D5, and the distance from the starting end of the mixed liquid outlet pipeline 41 to the second mixing point 50 is D6, satisfying that D5≤D4<D6; the neutralizing liquid supply The pipeline 30 is used to close the neutralizing liquid circuit 31 and open the neutralizing liquid supply pipeline 30 when the third flow detection element 42 detects that the cumulative flow of the preliminary mixed liquid is greater than 0, so that the neutralizing liquid in the neutralizing liquid supply pipeline 30 flows to the second mixing point 50. The neutralizing liquid that fills the neutralizing liquid self-circulation pipeline can respond quickly and flow to the second mixing point 50 at the same time or in advance relative to the preliminary mixed liquid in the mixed liquid outlet pipeline 41, thereby avoiding empty flow loss of the preliminary mixed liquid and inability to fully mix with the neutralizing liquid, thereby ensuring that the mixing reaction between the cell suspension, the lysate and the neutralizing liquid meets the ratio requirements; it can also take into account that under different flow rate ratios, the neutralizing liquid and the preliminary mixed liquid basically reach the second mixing point 50 at the same time or the neutralizing liquid reaches the first mixing point 40 slightly earlier than the preliminary mixed liquid, thereby reducing material waste.

[0094] Preferably, the relationship between the first flow detection element 11 and the lysate outlet point 28 and the first mixing point 40 also satisfies that D1+D2≤100mm, that is, under the premise of ensuring D2≤D1, the sum of the distance of the cell suspension from the first flow detection element 11 to the first mixing point 40 and the distance of the lysate from the lysate outlet point 28 to the first mixing point 40 is less than 100mm, ensuring that the cell suspension and the lysate flow to the first mixing point 40 as soon as possible and mix, thereby improving the mixing reaction efficiency; if D1+D2>100mm, and D2≤D1, the distance from the first flow detection element 11 to the first mixing point 40 is too short. The cell suspension itself has a greater fluid concentration and viscosity than the lysate, and its flow rate in the cell suspension supply pipeline 10 is slower than that of the lysate. The first flow detection element 11 detects the cell suspension and sends a signal to make the lysate flow out of the lysate outlet point 28. This may result in the cell suspension arriving at the first mixing point 40 a long time after the lysate arrives at the first mixing point 40 to mix with the lysate, resulting in a large amount of lysate waste. Therefore, controlling D1+D2≤100mm is beneficial to reducing material waste, improving mixing efficiency, and making the mixing reaction of the cell suspension and the lysate more in line with the ratio requirements.

[0095] Preferably, the relationship between the third flow detection element 42 and the neutralization liquid outlet point 37 and the second mixing point 50 also satisfies that D4+D5≤100mm, that is, under the premise of ensuring D5≤D4, the sum of the distance of the preliminary mixed liquid from the third flow detection element 42 to the second mixing point 50 and the distance of the neutralization liquid from the neutralization liquid outlet point 37 to the second mixing point 50 is less than 100mm, ensuring that the preliminary mixed liquid and the neutralization liquid flow to the second mixing point 50 as soon as possible and mix, thereby improving the mixing reaction efficiency; if D4+D5>100mm, and D5≤D4, then the distance from the third flow detection element 42 to the second mixing point 50 is too short. The preliminary mixed liquid has a fluid concentration and a viscosity greater than that of the neutralizing liquid, and its flow rate in the mixed liquid outlet pipeline 41 is slower than that of the neutralizing liquid. When the third flow detection element 42 detects the preliminary mixed liquid and sends a signal to make the neutralizing liquid flow out of the neutralizing liquid outlet point 37, it may result in the preliminary mixed liquid arriving at the second mixing point 50 a long time after the neutralizing liquid arrives at the second mixing point 50 for mixing with the neutralizing liquid, resulting in a large amount of waste of the neutralizing liquid. Therefore, controlling D4+D5≤100mm is beneficial to reducing material waste, improving mixing efficiency, and making the mixing reaction of the preliminary mixed liquid and the neutralizing liquid more in line with the ratio requirements.

[0096] like Figure 1 In the first embodiment shown, the system of the embodiment of the present invention further includes a control module (not shown), which can be a functional module or a program module, and can be implemented by software or hardware. For a module implemented by hardware, the control module is a processor or a part of a processor.

[0097] A first supply pump 12 is provided on the cell suspension supply pipeline 10, and the control module is communicatively connected to the first supply pump 12 for controlling the operation of the first supply pump 12, automatically controlling the first supply pump 12 to start, and pushing the cell suspension to flow toward the first mixing point 40 on the cell suspension supply pipeline 10; the first supply pump 12 is a peristaltic pump, which is wiredly or wirelessly connected to the control module.

[0098] A second supply pump 23 and a first valve 24 are provided on the lysate supply pipeline 20, and the first valve 24 is located between the lysate outlet point 28 and the first mixing point 40. A second valve 25 is provided on the lysate loop 22, and the control module is respectively connected to the second supply pump 23, the first valve 24, the second valve 25 and the first flow detection element 11 for communication, and is used to control the operation of the second supply pump 23, and is also used to open the second valve 25 and close the first valve 24 before the cumulative flow of the cell suspension detected by the first flow detection element 11 is greater than 0, so that the lysate flows in the lysate self-circulation pipeline, and is also used to close the second valve 25 and open the first valve 24 after the cumulative flow of the cell suspension detected by the first flow detection element 11 is greater than 0, so that the lysate flows out of the lysate outlet point 28 and flows to the first mixing point 40.

[0099] The control system of the present invention automatically controls the supply of the lysis solution through the control module, preferentially opens the second supply pump 23 and the second valve 25, so that the lysis solution fills the lysis solution self-circulation pipeline, and is ready to flow quickly from the lysis solution outlet point 28 to the first mixing point 40, and then after the first flow detection element 11 detects the cell suspension, that is, after the cell suspension is relatively close to the first mixing point 40, the second valve 25 is closed and the first valve 24 is opened, the lysis solution can directly flow out of the lysis solution outlet point 28 and quickly flow to the first mixing point 40, avoiding the cell suspension flowing to the first mixing point 40 first, resulting in part of the cell suspension being unable to mix with the lysis solution, causing cell suspension loss.

[0100] The second supply pump 23 is a peristaltic pump, and the first valve 24 and the second valve 25 are solenoid valves, which are all connected to the control module by wired communication or wireless communication.

[0101] A third supply pump 32 and a third valve 33 are provided on the neutralizing liquid supply pipeline 30. The third valve 33 is located between the neutralizing liquid outlet point 37 and the second mixing point 50. A fourth valve 34 is provided on the neutralizing liquid circuit 31. The control module is communicated and connected with the third supply pump 32, the third valve 33, the fourth valve 34 and the third flow detection element 42 respectively, and is used to control the operation of the third supply pump 32. It is also used to open the fourth valve 34 and close the third valve 33 before the cumulative flow of the preliminary mixed liquid detected by the third flow detection element 42 is greater than 0, so that the neutralizing liquid flows in the neutralizing liquid self-circulation pipeline. It is also used to close the fourth valve 34 and open the third valve 33 after the cumulative flow of the preliminary mixed liquid detected by the third flow detection element 42 is greater than 0, so that the neutralizing liquid flows out of the neutralizing liquid outlet point 37 and flows to the second mixing point 50.

[0102] The control system of the present invention automatically controls the supply of neutralizing liquid through the control module, and preferentially opens the third supply pump 32 and the fourth valve 34, so that the neutralizing liquid fills the neutralizing liquid self-circulating pipeline, and is ready to flow quickly from the neutralizing liquid outlet point 37 to the first mixing point 40. Then, after the third flow detection element 42 detects the preliminary mixed liquid, which means that the preliminary mixed liquid is relatively close to the second mixing point 50, the fourth valve 34 is closed and the third valve 33 is opened, so that the neutralizing liquid can directly flow out of the neutralizing liquid outlet point 37 and quickly flow to the second mixing point 50, thereby avoiding the preliminary mixed liquid flowing to the first mixing point 40 first, resulting in part of the preliminary mixed liquid being unable to mix with the neutralizing liquid, resulting in loss of the preliminary mixed liquid.

[0103] The third supply pump 32 is a peristaltic pump, and the third valve 33 and the fourth valve 34 are solenoid valves, which are all connected to the control module by wired communication or wireless communication.

[0104] like Figure 1In the first embodiment shown, a first mixing device is connected downstream of the first mixing point 40 of the embodiment of the present invention, and a first mixing section 45 is connected between the first mixing point 40 and the inlet end of the first mixing device. The first mixing point 40 is used for the initial meeting and mixing of the cell suspension and the lysate, and the first mixing device is used for the full mixing of the cell suspension and the lysate. The length of the first mixing section 45 determines the distance from the first mixing point 40 to the first mixing device. The length of the first mixing section 45 is d1, satisfying d1<100mm, so that the initially met cell suspension and lysate can be quickly moved to the first mixing device for uniform mixing; if the length of the first mixing section 45 is greater than 100mm, part of the cell suspension and the lysate are mixed and reacted in the first mixing section 45, and the concentration and viscosity of the feed fluid increase, which may block the first mixing section 45, resulting in the inability of the subsequent cell suspension and lysate to meet at the first mixing point 40 according to the set flow rate, affecting the timing accuracy of the entire control system.

[0105] The first mixing device includes a first spiral mixer 43 and a coil mixer 44. The cell suspension and the lysate are fully mixed in the first spiral mixer 43 and then fully reacted in the coil mixer 44 to form a preliminary mixed solution.

[0106] like Figure 1 In the first embodiment shown, a second mixing device is connected downstream of the second mixing point 50, and a second mixing section 52 is connected between the second mixing point 50 and the inlet end of the second mixing device. The second mixing point 50 is used for the preliminary meeting and mixing of the preliminary mixed liquid and the neutralizing liquid, and the second mixing device is used for the sufficient mixing of the preliminary mixed liquid and the neutralizing liquid. The length of the second mixing section 52 determines the distance from the second mixing point 50 to the second mixing device. The length of the second mixing section 52 is d2, satisfying d2<100mm, so that the preliminary mixed liquid and the neutralizing liquid that initially meet can be quickly moved to the second mixing device for uniform mixing; if the length of the second mixing section 52 is greater than 100mm, the preliminary mixed liquid itself is very viscous, and it cannot be quickly mixed with the neutralizing liquid in the second mixing section 52, and it is easy to block the second mixing section 52, resulting in the subsequent preliminary mixed liquid and the neutralizing liquid being unable to meet at the second mixing point 50 according to the set flow rate, affecting the timing control accuracy of the entire control system.

[0107] The second mixing device includes a second spiral mixer 51, in which the preliminary mixed liquid and the neutralizing liquid are fully mixed and reacted to form a final mixture; the final mixture is subsequently clarified and purified to obtain the target product plasmid.

[0108] The first spiral mixer 43, the coil mixer 44 and the second spiral mixer 51 can be conventional mixers in the art, and the specific structures are not described in detail here.

[0109] Based on different cell suspensions, the arrangements of the cell suspension supply module and the lysis solution supply module are also different.

[0110] like Figure 1 In the illustrated embodiment, since the cell suspension contains solids such as cells, the solid content of the cell suspension is high and the viscosity is large, and the flow speed is slow. The fluid concentration and viscosity of the lysate are small, and the flow rate is more controllable. Therefore, the cell suspension supply module and the lysate supply module are arranged in the longitudinal direction, with the cell suspension supply module on the top and the lysate supply module on the bottom. The cell suspension supply pipeline 10 includes a first transverse flow section 13 connected to the first mixing point 40, so that the cell suspension flows smoothly in the cell suspension supply pipeline 10, and is less affected by gravity, which is conducive to ensuring the stability and uniformity of the cell suspension liquid performance and flow rate; the lysate supply pipeline 20 includes a second transverse flow section 26 and a first longitudinal flow section 27, the second transverse flow section 26 is located below the first transverse flow section 13, and the first longitudinal flow section 27 connects the second transverse flow section 26 and the first mixing point 40 from bottom to top. The lysate flows smoothly in the second transverse flow section 26 and flows upward in the first longitudinal flow section 27. Since the adhesion between the lysate and the inner wall of the tube is small, it is easier to flow upward and control the flow rate.

[0111] In some embodiments, the fluid concentration and viscosity of the cell suspension are relatively small, or are close to the fluid concentration and viscosity of the lysate, and the cell suspension can be Figure 4 In the fourth embodiment shown, the cell suspension supply module and the lysis solution supply module are arranged longitudinally, with the cell suspension supply module at the bottom and the lysis solution supply module at the bottom. The cell suspension supply pipeline 10 includes a first transverse flow segment 13 and a third longitudinal flow segment 14, and the third longitudinal flow segment 14 is connected to the first mixing point 40 from bottom to top, and the lysis solution supply pipeline 20 includes a second transverse flow segment 26 and a first longitudinal flow segment 27, and the first longitudinal flow segment 27 is connected to the first mixing point 40 from top to bottom.

[0112] exist Figure 1 In the illustrated embodiment 1, the first supply pump 12 on the cell suspension supply pipeline 10 is arranged on the first lateral flow section 13, and the first flow detection element 11 is located upstream of the first supply pump 12. It takes a certain amount of time for the first flow detection element 11 to detect the cell suspension and send a signal to make the lysate flow out of the lysate outlet point 28. The first flow detection element 11 is arranged upstream of the first supply pump 12, so that the lysate receives the cell suspension presence signal early and is released in time and flows to the first mixing point 40, thereby avoiding the cell suspension flowing to the first mixing point 40 before the lysate and causing the loss of the cell suspension.

[0113] In other embodiments, Figure 3In the third embodiment shown, the first supply pump 12 on the cell suspension supply pipeline 10 is arranged on the first lateral flow section 13, and the first flow detection element 11 is located downstream of the first supply pump 12, and is close to the first mixing point 40. However, the flow speed of the cell suspension is slow, and sufficient communication time and feedback time are reserved to avoid the cell suspension flowing to the first mixing point 40 before the lysis solution, thereby preventing the cell suspension from losing.

[0114] Generally speaking, the solid content and viscosity of the preliminary mixed solution are greater than the fluid concentration and viscosity of the neutralized solution because the preliminary mixed solution contains solids such as cell fragments and biological products. Figure 1 In the illustrated embodiment 1, the mixed liquid outlet pipeline 41 of the first mixing point 40 and the second mixing point 50 are located above the neutralizing liquid supply module. The preliminary mixed liquid can use the gravity of the preliminary mixed liquid to promote the preliminary mixed liquid to flow more smoothly in the mixed liquid outlet pipeline 41 to the second mixing point 50. The neutralizing liquid supply pipeline 30 includes a third transverse flow section 35 and a second longitudinal flow section 36. The second longitudinal flow section 36 connects the third transverse flow section 35 and the second mixing point 50 from bottom to top. The neutralizing liquid flows smoothly in the third transverse flow section 35 and flows upward in the second longitudinal flow section 36. Since the adhesion between the neutralizing liquid and the inner wall of the tube is small, it is easier to flow upward and control the flow rate.

[0115] like Figure 1 In the illustrated embodiment 1, the second supply pump 23 on the lysate supply pipeline 20 is arranged on the second transverse flow section 26, and the second flow detection element 21 is located downstream of the second supply pump 23 and is arranged on the first longitudinal flow section 27. The second supply pump 23 is arranged on the second transverse flow section 26, which is conducive to the smooth flow of the lysate in the lysate supply pipeline 20, and the second flow detection element 21 is used to detect the cumulative flow of the lysate outflowing from the outlet point 28, and the first longitudinal flow section 27 is a flow section closer to the first mixing point 40, which ensures the accuracy of the second flow detection element 21 in detecting the cumulative flow of the lysate, thereby ensuring that the mixing reaction between the cell suspension and the lysate and the neutralization solution meets the ratio requirements, the cell suspension can be completely consumed without waste, and the batch consistency in the mass production process is also ensured.

[0116] Based on the control system of the above embodiment, the embodiment of the present invention further discloses a control method based on a cell lysis process, comprising the following steps:

[0117] Mixing step: opening the cell suspension supply pipeline 10, opening the lysis solution supply pipeline 20, and opening the neutralization solution supply pipeline 30;

[0118] The lysate reaches the first mixing point 40 before the cell suspension or reaches the first mixing point 40 at the same time;

[0119] The lysate and the cell suspension are mixed at a first mixing point 40 to form a preliminary mixed solution;

[0120] The neutralizing liquid reaches the second mixing point 50 before the preliminary mixed liquid or reaches the second mixing point 50 at the same time, and is mixed at the second mixing point 50;

[0121] In the mixing step, based on different pipeline lengths, the lysis solution can be opened first, then the cell suspension, and finally the neutralizing solution; or, the cell suspension can be opened first, then the lysis solution, and finally the neutralizing solution; or, all three can be opened at the same time; or the first two can be opened at the same time, and the neutralizing solution can be opened last. Users can set it according to their own ratios and pipeline lengths, as long as the lysis solution is controlled to reach the first mixing point 40 before the cell suspension or reach the first mixing point 40 at the same time to avoid the loss of the cell suspension caused by the inability of part of the cell suspension to contact and mix with the lysis solution, and the neutralizing solution is controlled to reach the second mixing point 50 before the preliminary mixed solution or reach the second mixing point 50 at the same time to avoid the loss of the preliminary mixed solution caused by the inability of part of the preliminary mixed solution to contact and mix with the neutralizing solution.

[0122] Emptying step: When the cumulative flow of the cell suspension detected by the first flow detection element 11 reaches the first flow setting value or the cumulative flow of the lysate detected by the second flow detection element 21 reaches the second flow setting value, the inert supplementary liquid supply module 60 supplies the inert supplementary liquid to the cell suspension supply pipeline 10, so that the cell suspension in the cell suspension supply pipeline 10 flows continuously until the inert supplementary liquid supply time is reached.

[0123] In the emptying step, the first flow setting value refers to the cumulative flow of the cell suspension when the cell suspension is close to being emptied, and the second flow setting value refers to the cumulative flow of the lysate when the lysate is mixed with most of the cell suspension in proportion, that is, the cumulative flow of the lysate is compared with the cumulative flow of the cell suspension. The first flow setting value or the second flow setting value can both indicate that the cell suspension is close to being emptied, and then the inert supplementary liquid supply module 60 is used to supply the inert supplementary liquid to the cell suspension supply pipeline 10, so that the cell suspension in the cell suspension supply pipeline 10 flows continuously until the cell lysis reaction is completely completed, thereby avoiding the cell suspension remaining in the cell suspension supply pipeline 10 and avoiding the preliminary mixed liquid remaining in the first mixing point 40, causing loss and waste.

[0124] More specifically, based on the control system of the above embodiment, the lysate supply module further includes a lysate loop 22 connected to the lysate supply pipeline 20 to form a lysate self-circulating pipeline, and the neutralizing liquid supply module further includes a neutralizing liquid loop 31 connected to the neutralizing liquid supply pipeline 30 to form a neutralizing liquid self-circulating pipeline; the mixing step also includes a starting step:

[0125] Open the lysate self-circulation pipeline so that the lysate fills the lysate self-circulation pipeline;

[0126] Open the neutralization liquid self-circulation pipeline so that the neutralization liquid fills the neutralization liquid self-circulation pipeline;

[0127] Open the cell suspension supply pipeline 10, and the cell suspension flows to the first flow detection element 11;

[0128] The mixing step also includes:

[0129] When the accumulated flow rate of the cell suspension detected by the first flow detection element 11 is greater than 0, the lysate loop 22 is closed, the lysate supply pipeline 20 is opened, and the lysate in the lysate supply pipeline 20 flows to the first mixing point 40, and the lysate is mixed with the cell suspension at the first mixing point 40;

[0130] When the cumulative flow of the preliminary mixed liquid detected by the third flow detection element 42 is greater than 0, the neutralizing liquid circuit 31 is closed, and the neutralizing liquid supply pipeline 30 is opened, so that the neutralizing liquid in the neutralizing liquid supply pipeline 30 flows to the second mixing point 50, and the neutralizing liquid is mixed with the preliminary mixed liquid at the second mixing point 50.

[0131] Corresponding to Figure 1 The specific components of the control method of one embodiment of the present invention are as follows:

[0132] Step 1: preset relevant parameters according to the recipe: pump speed v1 of the first supply pump 12, pump speed v2 of the second supply pump 23, and pump speed v3 of the third supply pump 32, a first flow setting value Q1, and Q1 is less than the processing volume of the cell suspension, a second flow setting value Q2, Q2 and Q1 meet the ratio requirements, and a preset time T;

[0133] Step 2: Open the second valve 25, the second supply pump 23, the fourth valve 34, and the third supply pump 32, keep the first valve 24 and the third valve 33 closed, and allow the lysate to flow in the lysate self-circulation pipeline, and the neutralization solution to flow in the neutralization solution self-circulation pipeline;

[0134] Step 3: Turn on the first supply pump 12 to start supplying the cell suspension;

[0135] Step 4: When the cumulative flow rate of the cell suspension detected by the first flow detection element 11 is greater than 0, the control module records the time point as the first time, closes the second valve 25, and opens the first valve 24 to allow the lysate to flow out of the lysate outlet 28. The cumulative flow rate of the lysate detected by the second flow detection element 21 is greater than 0, and the control module records the time point as the second time, and flows to the first mixing point 40. The lysate and the cell suspension meet at the first mixing point 40, and are fully mixed in the first spiral mixer 43, and fully react in the coil mixer 44 to form a preliminary mixed solution;

[0136] Step 5: The cumulative flow rate of the preliminary mixed liquid detected by the third flow detection element 42 is greater than 0, the control module records the time point as the third time, closes the fourth valve 34, opens the third valve 33, and the neutralizing liquid flows out of the neutralizing liquid outlet point 37 and flows to the second mixing point 50. The preliminary mixed liquid meets the neutralizing liquid at the second mixing point 50 and is fully mixed and reacted in the second spiral mixing liquid to form a final mixed liquid;

[0137] Step 6: When the accumulated flow rate of the cell suspension detected by the first flow detection element 11 reaches the first flow setting value, or the accumulated flow rate of the lysate detected by the second flow detection element 21 reaches the second flow setting value, the fifth valve 61 and the sixth valve 62 are opened, and the inert supplementary liquid supply module 60 supplies the inert supplementary liquid to the cell suspension supply pipeline 10 so that the cell suspension in the cell suspension supply pipeline 10 flows continuously, and supplies the inert supplementary liquid to the lysate supply pipeline 20 so that the lysate in the lysate supply pipeline 20 flows continuously;

[0138] It should be noted that step 6 does not necessarily start after step 5. It may also start before step 5 or be carried out at the same time as step 5.

[0139] Step 7: Count the supply time of the inert supplementary liquid supply module 60 until the difference between the third time and the first time, the difference between the third time and the second time, or the preset time T is reached, stop the first supply pump 12, the second supply pump 23 and the third supply pump 32, and the cell lysis reaction is completed.

[0140] The times in the above steps are all recorded by the control module and converted from system time to timestamps and recorded in the control module. For example, the time of step two is system time 8:00, the time of step three is system time 8:10, the first time is system time 8:15, the second time is system time 8:16, and the third time is system time 9:00. The preset time T is 45 minutes, wherein the difference between the third time and the first time is 45 minutes, and the difference between the third time and the second time is 44 minutes. Therefore, the inert replenishing liquid supply module 60 stops when the cumulative supply time is 44 minutes. At the same time, the first supply pump 12, the second supply pump 23 and the third supply pump 32 are stopped, thereby realizing a high degree of automation of the entire system, avoiding human operation errors or low intelligent automatic control of the liquid preparation system, thereby ensuring batch consistency during mass production.

[0141] The above-mentioned embodiments are only preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and substitutions made by technicians in this field on the basis of the present invention shall fall within the scope of protection required by the present invention.

Claims

1. A control system based on a cell lysis process, comprising a cell suspension supply module, a lysis solution supply module, a neutralization solution supply module, a first mixing point and a second mixing point, wherein a mixed solution outlet pipeline is connected between the first mixing point and the second mixing point; It is characterized in that The cell suspension supply module comprises a cell suspension supply pipeline, the lysate supply module comprises a lysate supply pipeline, and the lysate supply pipeline is connected with the cell suspension supply pipeline at the first mixing point; It also includes a first flow detection element, which is used to detect the cumulative flow of the cell suspension flowing through the cell suspension supply pipeline; And / or, further comprising a second flow detection element, for detecting the cumulative flow of the lysate flowing from the lysate supply pipeline into the first mixing point; The neutralizing liquid supply module comprises a neutralizing liquid supply pipeline, and the neutralizing liquid supply pipeline is connected with the mixed liquid outlet pipeline at the second mixing point; It also includes a third flow detection element, which is used to detect the cumulative flow of the preliminary mixed liquid flowing through the mixed liquid outlet pipeline; It also includes an inert supplementary liquid supply module connected to the cell suspension supply pipeline. The inert supplementary liquid supply module is used to supply inert supplementary liquid to the cell suspension supply pipeline to enable the cell suspension in the cell suspension supply pipeline to flow continuously.

2. The control system according to claim 1, characterized in that: It also includes an inert supplementary liquid time automatic control unit, which is communicatively connected with the inert supplementary liquid supply module and is used to control the supply time of the inert supplementary liquid.

3. The control system according to claim 1, characterized in that: The lysate supply module also includes a lysate loop connected to the lysate supply pipeline to form a lysate self-circulating pipeline, the connection point between the lysate supply pipeline and the lysate loop is defined as a lysate outlet point, the position of the second flow detection element on the lysate supply pipeline is downstream of the lysate outlet point, the distance from the first flow detection element to the first mixing point is D1, the distance from the lysate outlet point to the first mixing point is D2, and the distance from the starting end of the cell suspension supply pipeline to the first mixing point is D3, satisfying D2≤D1<D3; the lysate supply pipeline is used to close the lysate loop and open the lysate supply pipeline when the cumulative flow of the cell suspension detected by the first flow detection element is greater than 0, so that the lysate in the lysate supply pipeline flows to the first mixing point; and / or The neutralizing liquid supply module also includes a neutralizing liquid circuit connected to the neutralizing liquid supply pipeline to form a neutralizing liquid self-circulating pipeline, the connection between the neutralizing liquid supply pipeline and the neutralizing liquid circuit is defined as a neutralizing liquid outlet point, the distance from the third flow detection element to the second mixing point is D4, the distance from the neutralizing liquid outlet point to the second mixing point is D5, and the distance from the starting end of the mixed liquid outlet pipeline to the second mixing point is D6, satisfying D5≤D4<D6; the neutralizing liquid supply pipeline is used to close the neutralizing liquid circuit and open the neutralizing liquid supply pipeline when the third flow detection element detects that the cumulative flow of the preliminary mixed liquid is greater than 0, so that the neutralizing liquid in the neutralizing liquid supply pipeline flows to the second mixing point.

4. The control system according to claim 3, characterized in that: The relationship between the first flow detection element, the lysate outlet point and the first mixing point also satisfies: D1+D2≤100mm; and / or The relationship between the third flow detection element, the neutralization liquid outlet point and the second mixing point also satisfies that D4+D5≤100mm.

5. The control system according to claim 3, characterized in that: It also includes a control module, the cell suspension supply pipeline is provided with a first supply pump, the control module is in communication connection with the first supply pump, and is used to control the operation of the first supply pump; and / or The invention also includes a control module, wherein a second supply pump and a first valve are provided on the lysate supply pipeline, the first valve is located between the lysate outlet point and the first mixing point, and a second valve is provided on the lysate loop. The control module is respectively connected to the second supply pump, the first valve, the second valve and the first flow detection element for communication, and is used to control the operation of the second supply pump, and is also used to open the second valve and close the first valve before the cumulative flow of the cell suspension detected by the first flow detection element is greater than 0, so that the lysate flows in the lysate self-circulation pipeline, and is also used to close the second valve and open the first valve after the cumulative flow of the cell suspension detected by the first flow detection element is greater than 0, so that the lysate flows out of the lysate outlet point and flows to the first mixing point; and / or It also includes a control module. A third supply pump and a third valve are provided on the neutralization liquid supply pipeline. The third valve is located between the neutralization liquid outlet point and the second mixing point. A fourth valve is provided on the neutralization liquid circuit. The control module is communicatively connected with the third supply pump, the third valve, the fourth valve and the third flow detection element respectively, for controlling the operation of the third supply pump, and for opening the fourth valve and closing the third valve before the cumulative flow of the preliminary mixed liquid detected by the third flow detection element is greater than 0, so that the neutralization liquid flows in the neutralization liquid self-circulation pipeline, and for closing the fourth valve and opening the third valve after the cumulative flow of the preliminary mixed liquid detected by the third flow detection element is greater than 0, so that the neutralization liquid flows out of the neutralization liquid outlet point and flows to the second mixing point.

6. The control system according to claim 1, characterized in that: The inert supplementary liquid supply module is also connected to the lysate supply pipeline, and is used to supply the inert supplementary liquid to the lysate supply pipeline, so that the lysate in the lysate supply pipeline flows continuously.

7. The control system according to claim 6, characterized in that: A fifth valve is provided between the inert supplementary liquid supply module and the cell suspension supply pipeline, and a sixth valve is provided between the inert supplementary liquid supply module and the lysate supply pipeline.

8. The control system according to claim 1, characterized in that: A first mixing device is connected downstream of the first mixing point, a first mixing section is connected between the first mixing point and the inlet end of the first mixing device, and the length of the first mixing section is d1, satisfying d1≤100 mm; and / or A second mixing device is connected downstream of the second mixing point, a second mixing section is connected between the second mixing point and the inlet end of the second mixing device, and the length of the second mixing section is d2, satisfying d2≤100 mm.

9. The control system according to claim 8, characterized in that: The first mixing device comprises a first spiral mixer and a coil mixer; and / or The second mixing device includes a second spiral mixer.

10. The control system according to claim 1, characterized in that: The cell suspension supply module and the lysis solution supply module are arranged in the longitudinal direction, the cell suspension supply pipeline includes a first transverse flow segment connected to the first mixing point, the lysis solution supply pipeline includes a second transverse flow segment and a first longitudinal flow segment, the second transverse flow segment is located below the first transverse flow segment, and the first longitudinal flow segment connects the second transverse flow segment and the first mixing point from bottom to top; and / or The mixed liquid outlet pipeline of the first mixing point and the second mixing point are located above the neutralizing liquid supply module, and the neutralizing liquid supply pipeline includes a third transverse flow segment and a second longitudinal flow segment, and the second longitudinal flow segment connects the third transverse flow segment and the second mixing point from bottom to top.

11. The control system according to claim 10, characterized in that: The first supply pump on the cell suspension supply pipeline is arranged on the first lateral flow section, and the first flow detection element is located upstream of the first supply pump; and / or The second supply pump on the lysate supply pipeline is arranged on the second transverse flow section, and the second flow detection element is located downstream of the second supply pump and is arranged on the first longitudinal flow section.

12. A control method based on a cell lysis process, characterized in that: The control system according to any one of claims 1 to 11 comprises the following steps: Mixing step: opening the cell suspension supply line, opening the lysis solution supply line, and opening the neutralization solution supply line; The lysate reaches the first mixing point before the cell suspension or reaches the first mixing point at the same time; The lysate and the cell suspension are mixed at the first mixing point to form a preliminary mixed solution; The neutralizing liquid reaches the second mixing point before the preliminary mixed liquid or reaches the second mixing point at the same time, and is mixed at the second mixing point; Emptying step: When the cumulative flow of the cell suspension detected by the first flow detection element reaches a first flow setting value or the cumulative flow of the lysate detected by the second flow detection element reaches a second flow setting value, the inert supplementary liquid supply module supplies an inert supplementary liquid to the cell suspension supply pipeline to allow the cell suspension in the cell suspension supply pipeline to flow continuously until the inert supplementary liquid supply time is reached.

13. The control method according to claim 12, characterized in that: The time when the cumulative flow rate of the cell suspension detected by the first flow detection element is greater than 0 is defined as the first time, the time when the cumulative flow rate of the lysate detected by the second flow detection element is greater than 0 is defined as the second time, and the time when the cumulative flow rate of the preliminary mixed solution detected by the third flow detection element is greater than 0 is defined as the third time; In the emptying step, the inert replenishing liquid supply time of the inert replenishing liquid supply module is determined according to the difference between the third time and the first time; or, the inert replenishing liquid supply time of the inert replenishing liquid supply module is determined according to the difference between the third time and the second time; or, the inert replenishing liquid supply time of the inert replenishing liquid supply module is executed according to a preset time.

14. The control method according to claim 12, characterized in that: The lysate supply module further comprises a lysate loop connected to the lysate supply pipeline to form a lysate self-circulation pipeline, and the neutralization liquid supply module further comprises a neutralization liquid loop connected to the neutralization liquid supply pipeline to form a neutralization liquid self-circulation pipeline. The mixing step also includes a starting step: Open the lysate self-circulation pipeline so that the lysate fills the lysate self-circulation pipeline; Open the neutralization liquid self-circulation pipeline so that the neutralization liquid fills the neutralization liquid self-circulation pipeline; Opening the cell suspension supply pipeline, so that the cell suspension flows to the first flow detection element; The mixing step further comprises: When the accumulated flow rate of the cell suspension detected by the first flow detection element is greater than 0, closing the lysate loop, opening the lysate supply pipeline, allowing the lysate in the lysate supply pipeline to flow to the first mixing point, and the lysate is mixed with the cell suspension at the first mixing point; When the cumulative flow of the preliminary mixed liquid detected by the third flow detection element is greater than 0, the neutralizing liquid circuit is closed, the neutralizing liquid supply pipeline is opened, and the neutralizing liquid in the neutralizing liquid supply pipeline flows to the second mixing point, and the neutralizing liquid is mixed with the preliminary mixed liquid at the second mixing point.

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