Control device based on cell lysis process

Abstract

The utility model discloses a control device based on a cell lysis process, which comprises an equipment main body, a cell suspension supply module, a lysis solution supply module, a neutralizing solution supply module, a first mixing point and a second mixing point, and a mixed solution outlet pipeline is connected between the first mixing point and the second mixing point; at least one of the cell suspension supply pipeline, the lysis solution supply pipeline and the neutralizing solution supply pipeline is fixedly connected with the equipment main body; the system further comprises a first flow detection element, a second flow detection element and a third flow detection element, the first flow detection element is arranged on the cell suspension supply pipeline, the second flow detection element is arranged on the lysis solution supply pipeline, and the third flow detection element is arranged on the mixed solution outlet pipeline; the inert replenishing liquid supply module is connected with the cell suspension supply pipeline and is used for supplying inert replenishing liquid to the cell suspension supply pipeline, so that the cell suspension is completely consumed as much as possible, and loss is avoided.

Description

TECHNICAL FIELD

[0001] The utility model relates to cell lysis technical field, especially based on cell lysis process's control device. BACKGROUND

[0002] Cell lysis technology is a conventional means to obtain internal substances of cells, generally including biological products to be lysed, lysate and neutralizing liquid, such as alkaline lysis method for extracting plasmid DNA; and the biological products to be lysed generally use animal, plant or microbial cell suspension, and the lysate and neutralizing liquid use chemical solution, which needs to be mixed with the cell suspension and the lysate before use, and then the neutralizing liquid is mixed with the preliminary mixed product again to form a lysed mixed product. The lysed mixture generally includes cell lysate, protein and its lysate, nucleic acid and its lysate, organelle and its lysate, other metabolites and the mixture formed by the reaction between the above-mentioned substances and the processing agent used, wherein the target product can be any one or a mixture of several of the above-mentioned substances (in the current biological field, the target product obtained from animal cells is mostly antibody, the target product obtained from plant cells is mostly flavonoid alkaloid, and the target product obtained from microbial cells, especially Escherichia coli, is plasmid), and the impurities are the cell lysate, protein and its lysate, nucleic acid and its lysate, organelle and its lysate, other metabolites and the mixture formed by the reaction between the above-mentioned substances and the processing agent used except the target product.

[0003] At present, due to the different fluid concentrations and viscosities of the cell suspension, lysate, preliminary mixed product and neutralizing liquid, and the need to mix the materials according to the ratio during the cell lysis process, the feeding flow rates of the materials may be different, and at the same time, the personnel operation error or low intelligent automatic control of the liquid preparation system in the quantitative industry causes the mixing reaction between the cell suspension and the lysate and the neutralizing liquid to be insufficient or excessive stirring and mixing, and does not meet the ratio requirement, thereby causing a certain loss of the yield and recovery rate of the target product, and it is difficult to ensure the batch-to-batch consistency problem of multiple batches of production.

[0004] Therefore, it is necessary to design a control device based on the cell lysis process to solve the problem of batch-to-batch consistency of the cell suspension under the condition of minimum loss in the prior art. UTILITY MODEL CONTENTS

[0005] In view of the deficiencies in the prior art, the purpose of the utility model is to provide a control device based on the cell lysis process, which solves the problems of loss of cell suspension and batch-to-batch consistency in the existing cell lysis process.

[0006] In order to achieve the above-mentioned purpose, the utility model adopts the following technical scheme:

[0007] A control device based on a cell lysis process, comprising a device main body, a cell suspension supply module, a lysis liquid supply module, a neutralization liquid supply module, and a first mixing point and a second mixing point, a mixed liquid outlet pipeline being connected between the first mixing point and the second mixing point;

[0008] The cell suspension supply module comprises a cell suspension supply pipeline, the lysis liquid supply module comprises a lysis liquid supply pipeline, and the lysis liquid supply pipeline is in communication with the cell suspension supply pipeline at the first mixing point; the neutralization liquid supply module comprises a neutralization liquid supply pipeline, and the neutralization liquid supply pipeline is in communication with the mixed liquid outlet pipeline at the second mixing point;

[0009] At least one of the cell suspension supply pipeline, the lysis liquid supply pipeline, and the neutralization liquid supply pipeline is fixedly connected to the device main body;

[0010] Further comprising a first flow detection element, a second flow detection element, and a third flow detection element, the first flow detection element being arranged on the cell suspension supply pipeline, the second flow detection element being arranged on the lysis liquid supply pipeline, and the third flow detection element being arranged on the mixed liquid outlet pipeline;

[0011] Further comprising an inert supplement liquid supply module connected to the cell suspension supply pipeline, the inert supplement liquid supply module being used for supplying inert supplement liquid to the cell suspension supply pipeline to continuously flow the cell suspension in the cell suspension supply pipeline.

[0012] The control device is characterized in that the first flow detection element is arranged on the cell suspension supply pipeline to detect the cumulative flow of the cell suspension and to control the relevant process flow response, the second flow detection element is arranged on the lysate supply pipeline to detect the cumulative flow of the lysate and to control the relevant process flow response, and the third flow detection element is arranged on the mixed liquid outlet pipeline between the first mixing point and the second mixing point to detect the flow condition of the preliminary mixed liquid formed by the cell suspension and the lysate and to control the relevant process flow response.

[0013] Preferably, the control device further comprises an inert supplement liquid time automatic control unit, which is in communication connection with the inert supplement liquid supply module and is used for controlling the supply time of the inert supplement liquid.

[0014] Preferably, the lysis solution supply module further comprises a lysis solution return circuit in communication with the lysis solution supply pipeline to form a lysis solution self-circulation pipeline, the connection of the lysis solution supply pipeline and the lysis solution return circuit defines a lysis solution outflow point, the position of the second flow detection element on the lysis solution supply pipeline is downstream of the lysis solution outflow point, the distance from the first flow detection element to the first mixing point is D1, the distance from the lysis solution outflow 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.

[0015] The lysis solution return circuit and the lysis solution supply pipeline are in communication to form a lysis solution self-circulation pipeline, so that the lysis solution can be released in advance to fill the lysis solution self-circulation pipeline, the lysis solution outflow 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 or 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 outflow point to the first mixing point, so that when the lysis solution needs to be pushed to the first mixing point, the lysis solution filling the lysis solution self-circulation pipeline can be quickly released, and the cell suspension in the cell suspension supply pipeline can flow to the first mixing point at the same time or in advance, avoiding loss of the cell suspension due to empty flow; and the second flow detection element is arranged downstream of the lysis solution outflow point, so as to accurately detect the cumulative flow of the lysis solution reacted with the cell suspension, ensuring that the mixing reaction between the cell suspension and the lysis solution and the neutralization solution meets the ratio requirement.

[0016] Preferably, the neutralization solution supply module further comprises a neutralization solution return circuit in communication with the neutralization solution supply pipeline to form a neutralization solution self-circulation pipeline, the connection of the neutralization solution supply pipeline and the neutralization solution return circuit defines a neutralization solution outflow point, the distance from the third flow detection element to the second mixing point is D4, the distance from the neutralization solution outflow point to the second mixing point is D5, and the distance from the starting end of the mixed solution outflow pipeline to the second mixing point is D6, satisfying D5≤D4<D6.

[0017] The neutralization solution return circuit and the neutralization solution supply pipeline are in communication to form a neutralization solution self-circulation pipeline, so that the neutralization solution can be released in advance to fill the neutralization solution self-circulation pipeline, the neutralization solution outflow point is closer to the second mixing point relative to the starting end of the neutralization solution supply pipeline, and the distance from the third flow detection element or the starting end of the mixed solution outflow pipeline to the second mixing point is greater than the distance from the neutralization solution outflow point to the second mixing point, so that when the neutralization solution needs to be pushed to the second mixing point, the neutralization solution filling the neutralization solution self-circulation pipeline can be quickly released, and the preliminary mixed solution in the mixed solution outflow pipeline can flow to the second mixing point faster, avoiding loss of the preliminary mixed solution due to empty flow, ensuring that the mixing reaction between the cell suspension and the lysis solution and the neutralization solution meets the ratio requirement.

[0018] Preferably, the device body is provided with a control module, the cell suspension supply pipeline is provided with a first supply pump, and the control module is in communication connection with the first supply pump; the control module is arranged in the device body and is closer to the first supply pump, so that the signal is stronger and the automatic control of the first supply pump is more favorable.

[0019] Preferably, the device body is provided with a control module, the lysate supply pipeline is provided with a second supply pump and a first valve, the position of the second supply pump on the lysate supply pipeline is upstream of the lysate outlet point, the first valve is located between the lysate outlet point and the first mixing point, the lysate circuit is provided with a second valve, and the control module is in communication connection with the second supply pump, the first valve, the second valve and the first flow detection element.

[0020] The control module is arranged in the device body and is closer to the second supply pump, the first valve, the second valve and the first flow detection element, so that the signal is stronger, the signal transmission speed is faster, the automatic control is more precise and timely, and the second supply pump is located upstream of the lysate outlet point. The second supply pump and the second valve are preferentially opened, so that the lysate fills the lysate self-circulation pipeline and is ready to flow quickly from the lysate outlet point to the first mixing point. After the first flow detection element detects the cell suspension, it means that the cell suspension is relatively close to the first mixing point. Then, the second valve is closed and the first valve is opened. The lysate can flow out of the lysate outlet point and quickly flow to the first mixing point, so that the cell suspension cannot flow to the first mixing point first and cause the loss of part of the cell suspension.

[0021] Preferably, the device body is provided with a control module, the neutralizing liquid supply pipeline is provided with a third supply pump and a third valve, the position of the third supply pump on the neutralizing liquid supply pipeline is upstream of the neutralizing liquid outlet point, the third valve is located between the neutralizing liquid outlet point and the second mixing point, the neutralizing liquid circuit is provided with a fourth valve, and the control module is in communication connection with the third supply pump, the third valve, the fourth valve and the third flow detection element.

[0022] The control module is arranged in the device body, is closer to the third supply pump, the third valve, the fourth valve and the third flow detection element, has stronger signal and faster signal transmission speed, is favorable for accurate and timely automatic control, and the third supply pump is located at the upstream of the neutralizing liquid outlet point. The third supply pump and the fourth valve are preferentially opened, the neutralizing liquid fills the neutralizing liquid self-circulation pipeline, and preparation for the neutralizing liquid to flow quickly from the neutralizing liquid outlet point to the first mixing point is made. After the third flow detection element detects the preliminary mixed liquid, the fourth valve is closed and the third valve is opened. The neutralizing liquid can flow out of the neutralizing liquid outlet point and flow quickly to the second mixing point, so that the preliminary mixed liquid is prevented from flowing to the first mixing point first and causing part of the preliminary mixed liquid to fail to mix with the neutralizing liquid, thereby causing loss of the preliminary mixed liquid.

[0023] Preferably, the first flow detection element is arranged upstream of the first supply pump on the cell suspension supply pipeline. Although the first flow detection element is closer to the control module in the device body, it takes a certain time for the first flow detection element to detect the cell suspension and send a signal to make the lysing liquid flow out of the lysing liquid outlet point. Arranging the first flow detection element upstream of the first supply pump makes the distance between the first flow detection element and the first mixing point more appropriate, so that the lysing liquid receives the cell suspension signal early and is released in time to flow to the first mixing point, thereby preventing the cell suspension from flowing to the first mixing point before the lysing liquid and causing loss of the cell suspension.

[0024] Preferably, the cell suspension supply module and the lysing liquid supply module are arranged in a longitudinal direction. The cell suspension supply pipeline includes a first horizontal flow section connected to the first mixing point. The lysing liquid supply pipeline includes a second horizontal flow section and a first longitudinal flow section. The second horizontal flow section is located below the first horizontal flow section, and the first longitudinal flow section connects the second horizontal flow section and the first mixing point from bottom to top.

[0025] The cell suspension contains cells and other solids, so the solid content of the cell suspension is high and the viscosity is large, and the flow speed 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 favorable for ensuring the stability and uniformity of the cell suspension liquid performance and flow rate. The fluid concentration and viscosity of the lysing liquid are small, and the flow rate is more controllable. Therefore, the lysing liquid supply pipeline includes a second horizontal flow section arranged horizontally and a first longitudinal flow section arranged vertically and connecting the second horizontal flow section and the first mixing point. The lysing liquid flows smoothly in the second horizontal flow section and flows upward in the first longitudinal flow section. Due to the small adhesion of the lysing liquid to the inner wall of the pipeline, the lysing liquid is more likely to flow upward and the flow rate is more controllable.

[0026] Preferably, the mixed liquid outlet pipeline of the first mixing point and the second mixing point are located above the neutralizing liquid supply module, the neutralizing liquid supply pipeline comprises a third horizontal flow section and a second vertical flow section, and the second vertical flow section connects the third horizontal flow section and the second mixing point from bottom to top.

[0027] The primary mixed liquid contains cell fragments and biological products, and has a solid content and viscosity greater than that 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, so that the gravity of the primary mixed liquid can facilitate the smooth flow of the primary mixed liquid in the mixed liquid outlet pipeline to the second mixing point. The fluid concentration and viscosity of the neutralizing liquid are small, and the flow rate is more controllable. Therefore, the neutralizing liquid supply pipeline comprises a third horizontal flow section arranged horizontally and a second vertical flow section arranged vertically and connecting the third horizontal flow section and the second mixing point. The neutralizing liquid flows smoothly in the third horizontal flow section and flows upward in the second vertical flow section. Due to the small adhesion of the neutralizing liquid to the inner wall of the pipeline, the neutralizing liquid is more likely to flow upward and the flow rate is more controllable.

[0028] Preferably, the first supply pump on the cell suspension supply pipeline is arranged on the first horizontal flow section. The flow rate of the cell suspension on the first horizontal flow section is stable, and the first supply pump can more stably control the flow rate of the cell suspension.

[0029] Preferably, the second supply pump on the lysis liquid supply pipeline is arranged on the second horizontal flow section, and the second flow detection element is arranged on the first vertical flow section. The second supply pump arranged on the second horizontal flow section facilitates the smooth flow of the lysis liquid in the lysis liquid supply pipeline. The second flow detection element detects the cumulative flow rate of the lysis liquid flowing out of the lysis liquid outlet point. The first vertical flow section is closer to the first mixing point, ensuring the accuracy of the detection of the cumulative flow rate of the lysis liquid by the second flow detection element, so that the mixing reaction between the cell suspension and the lysis liquid and the neutralizing liquid meets the proportioning requirements, the cell suspension is completely consumed as much as possible, and the batch consistency in the mass production process is ensured.

[0030] Preferably, the equipment body comprises four vertically arranged side surfaces, which are divided into a first side surface, a second side surface, a third side surface and a fourth side surface.

[0031] Part of the cell suspension supply pipeline is arranged on the first side surface, and the first flow detection element is arranged on the first side surface. Part of the cell suspension supply pipeline is arranged on the second side surface, and the first supply pump is arranged on the second side surface. And / or

[0032] Part of the lysis solution supply pipeline is arranged on the first side, part of the lysis solution supply pipeline is arranged on the second side, and the second supply pump and the second flow detection element are arranged on the second side; and / or

[0033] Part of the neutralization solution supply pipeline is arranged on the third side, and the third supply pump and the third flow detection element are arranged on the third side.

[0034] The cell suspension supply pipeline, the lysis solution supply pipeline and the neutralization solution supply pipeline are all arranged on the side of the equipment body, making full use of the installation space of the side of the equipment body, and also making the components on each supply pipeline closer to the control module, which is conducive to signal transmission; the cell suspension supply pipeline and the lysis solution supply pipeline have stronger relevance, and are therefore arranged together on the first side and the second side, which is convenient for manual observation; the neutralization solution supply pipeline and the mixed solution outflow pipeline are arranged on the same side, have stronger relevance, and are convenient for manual observation, so that automatic control failures or other problems can be found in time.

[0035] Preferably, the mixed solution outflow pipeline defines an additional pipeline corresponding to the part between the third flow detection element and the second mixing point, a hanging bracket is arranged on the third side, and the additional pipeline is wound and arranged on the hanging bracket.

[0036] The fluid concentration and viscosity of the preliminary mixed solution formed by different types of cell suspensions are different, which results in different flow rates of the preliminary mixed solution in the mixed solution outflow pipeline, and the distance from the third flow detection element to the second mixing point also needs to be adjusted to make the neutralization solution and the preliminary mixed solution reach the second mixing point at the same time as much as possible. Therefore, the additional pipeline is adopted to facilitate the user to adjust the distance from the third flow detection element to the second mixing point based on actual needs, and the additional pipeline wound and arranged on the hanging bracket is used to make full use of the space on the equipment body, so that the pipeline arrangement is clearer and neater.

[0037] Preferably, the inert supplement solution supply module is also connected with the lysis solution supply pipeline, for supplying inert supplement solution to the lysis solution supply pipeline to make the lysis solution in the lysis solution supply pipeline flow continuously;

[0038] The fifth valve is arranged between the inert supplement solution supply module and the cell suspension supply pipeline, and the sixth valve is arranged between the inert supplement solution supply module and the lysis solution supply pipeline.

[0039] The role of the inert supplement solution in promoting the lysis solution is to reduce the use of lysis solution and avoid the lysis solution remaining in the lysis solution supply pipeline, and to balance the acid-base value of the preliminary mixed solution and avoid too much lysis solution flowing into the first mixing point to mix with the cell suspension and destroy the set mixing ratio.

[0040] Compared with the prior art, the utility model has at least the following beneficial effects:

[0041] The control device of the utility model, first flow detection element is established on cell suspension supply pipeline, detects the cumulative flow of cell suspension and controls the response of relevant process flow, second flow detection element is established on lysate supply pipeline, detects the cumulative flow of lysate and controls the response of relevant process flow, third flow detection element is established on mixed liquid outlet pipeline between first mixing point and second mixing point, detects the flow condition of the preliminary mixed liquid formed by cell suspension and lysate and controls the response of relevant process flow, thereby controls the flow condition of each liquid and mixed liquid of each stage in the whole cell lysis process, so that the mixing ratio between cell suspension and lysate and neutralizing liquid meets the matching requirement, cell suspension can be consumed completely as far as possible, waste is avoided, and batch consistency in mass production process is also ensured, at the same time, inert supplement liquid supply module is established, inert supplement liquid is supplied to cell suspension supply pipeline, inert supplement liquid does not react with cell suspension, lysate and neutralizing liquid, its role not only promotes the flow of cell suspension in cell suspension supply pipeline to first mixing point, avoids the residual of cell suspension in cell suspension supply pipeline, cell suspension can be consumed completely, loss is avoided, but also avoids the residual of preliminary mixed liquid in first mixing point, causes loss and waste, and then promotes the mixing of preliminary mixed liquid and neutralizing liquid. BRIEF DESCRIPTION OF DRAWINGS

[0042] In order to more clearly illustrate the specific embodiment of the utility model or the technical scheme in the prior art, the following will briefly introduce the drawings needed to be used in the specific embodiment or the prior art description, obviously, the drawings in the following description are some embodiments of the utility model, and for those skilled in the art, other drawings can also be obtained according to these drawings without creative labor.

[0043] Figure 1 It is the frame schematic view of the control device of the utility model embodiment one.

[0044] Figure 2 It is the structure schematic view of the control device of the utility model embodiment one.

[0045] Figure 3 It is another angle structure schematic view of the control device of the utility model embodiment one.

[0046] Figure 4 It is frame schematic view of the control device of the utility model embodiment two.

[0047] Figure 5 It is frame schematic view of the control device of the utility model embodiment three.

[0048] Figure 6 It is frame schematic view of the control device of the utility model embodiment four.

[0049] Figure 7 It is structure schematic view of the control device of the utility model embodiment five.

[0050] Explanation of reference signs

[0051] 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;

[0052] 20, lysate supply pipeline; 21, second flow detection element; 22, lysate circuit; 23, second supply pump; 24, first valve; 25, second valve; 26, second transverse flow section; 27, first longitudinal flow section; 28, lysate outlet point; 29, lysate bag;

[0053] 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;

[0054] 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; 46, additional pipeline;

[0055] 50, second mixing point; 51, second spiral mixer; 52, second mixing section; 53, collection bag;

[0056] 60, inert supplement liquid supply module; 61, fifth valve; 62, sixth valve; 63, inert supplement liquid storage tank;

[0057] 70, equipment main body; 71, first side; 72, second side; 73, third side; 74, fourth side; 75, hanging support. DETAILED DESCRIPTION

[0058] The technical solutions of the present application will be described clearly and completely below in conjunction with the drawings. Obviously, the described embodiments are some of the embodiments of the present application, but not all the embodiments. Based on the embodiments of the present application, all the other embodiments obtained by those skilled in the art without creative efforts belong to the scope of protection of the present application.

[0059] In the description of the present application, it should be noted that the orientations or positional relationships indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. are based on the orientations or positional relationships shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and therefore cannot be understood as indicating or implying that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the present application. In addition, the terms "first", "second", "third" are only for the purpose of description, and cannot be understood as indicating or implying relative importance.

[0060] In the description of the present application, it should be noted that unless otherwise explicitly specified and limited, the terms "mounting", "connection", "connection" should be understood broadly, for example, it can be fixedly connected, or it can be detachably connected, or integrally connected; it can be mechanically connected, or it can be electrically connected; it can be directly connected, or it can be indirectly connected through an intermediate medium; it can be the communication inside two elements. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0061] The general process of cell lysis is to mix the cell suspension with the lysis solution sufficiently, so that the cells in the cell suspension are lysed, and the lysis mixture in the cells is released. The lysis mixture usually includes cell lysate, protein and its lysate, nucleic acid and its lysate, organelle and its lysate, other metabolites, etc. and the mixture formed by the reaction between the above-mentioned substances and the processing agent used, wherein the target product can be any one or a mixture of several of the above-mentioned substances (in the current biological field, the target product obtained from animal cells is mostly antibody, the target product obtained from plant cells is mostly flavonoid alkaloid, and the target product obtained from microbial cells, especially Escherichia coli, is plasmid), and the impurities are the cell lysate, protein and its lysate, nucleic acid and its lysate, organelle and its lysate, other metabolites, etc. and the mixture formed by the reaction between the above-mentioned substances and the processing agent used, except the target product.

[0062] The alkaline lysis plasmid extraction method is based on the difference between the denaturation and renaturation of chromosomal DNA and plasmid DNA to achieve the separation purpose. Therefore, the method generally includes three steps: 1. A certain proportion of resuspension liquid is provided after resuspension (containing E. coli), and a lysis solution (currently mainly using an alkaline reagent solution: such as 0.2M NaOH and 1% SDS) is added to the bacterial liquid, so that the lysis solution and the bacterial liquid are mixed quickly and fully, in this process, a large number of hydrogen bonds of chromosomal DNA are broken, and the double helix structure is opened and denatured, a small part of the hydrogen bonds of plasmid DNA are also broken, but the two complementary chains of the supercoiled covalent closed loop will not be completely separated; 2. After the bacterial liquid and the lysis solution are mixed and reacted, the lysis product generated includes the target plasmid and the bacterial lysis body, protein and its lysis body, chromosomal DNA and its lysis body, RNA, organelle and its lysis body, other metabolites and the like, and the complexes or mixtures formed between the above-mentioned substances and the treatment agents used; 3. When the acid salt solution is added for neutralization, the supercoiled plasmid DNA is renatured, and at the same time, the high concentration of KAC also plays a role of salting out, so that the formed SDS-gDNA-protein insoluble complex is precipitated, and a part of the denatured protein and lipid also become insoluble curd-like precipitate, while the plasmid DNA is still in a dissolved state, and the effective separation of the target product is realized by solid-liquid separation.

[0063] At present, in the cell lysis process, each material needs to be mixed according to the proportion, the fluid concentration and the viscosity of the cell suspension, the lysis solution, the preliminary mixed product and the neutralizing liquid are different, so that the feeding flow rate of each material is different, therefore, the control device of the embodiment of the utility model is improved, and the flow condition of each liquid in the whole cell lysis process can be regulated.

[0064] As Figures 1 to 3In the first embodiment shown, the control device based on the cell lysis process of the utility model embodiment, including equipment main body 70, cell suspension supply module, lysis liquid supply module, neutralization liquid supply module and first mixing point 40, second mixing point 50, first mixing point 40 and second mixing point 50 are connected with mixed liquid outlet pipeline 41.Equipment main body 70 is the installation basis of control device, at least one of cell suspension supply pipeline 10, lysis liquid supply pipeline 20 and neutralization liquid supply pipeline 30 is fixedly connected with equipment main body 70, the supply module not fixedly connected with equipment main body 70 is also arranged around equipment main body 70, first mixing point 40, second mixing point 50 can be arranged on equipment main body 70, also can be arranged on the storage device around equipment main body 70, keep close distance with equipment main body 70, facilitate pipeline connection and signal transmission;It is favorable to the rapid transmission of detection information and the efficient automatic control, more accurate detection, more can reflect the real-time flow condition of each liquid and each stage mixed liquid in cell lysis process.

[0065] Cell suspension supply module corresponds to the communication cell suspension bag 15 or cell suspension cylinder and other cell suspension storage devices, lysis liquid supply module corresponds to the communication lysis liquid bag 29 or lysis liquid cylinder and other lysis liquid storage devices, neutralization liquid supply module communicates neutralization liquid bag 38 or neutralization liquid cylinder and other neutralization liquid storage devices, these storage devices all include inlet valve and outlet valve, control material to flow out or flow into corresponding storage device.Storage device can also be fixedly connected with equipment main body 70, can also be arranged on the storage device around equipment main body 70, and the storage device can be shelf, hanging rack, trolley and other equipment, can also be wall or ground and other positions. Figure 2 And Figure 3 In the first embodiment shown, the cell suspension storage device, lysis liquid storage device and neutralization liquid storage device are all storage bags, and are hung near the equipment main body 70, that is, the cell suspension bag 15, the lysis liquid bag 29 and the neutralization liquid bag 38.

[0066] The cell suspension supply module comprises a cell suspension supply pipeline 10, which is in communication with an outlet valve of a cell suspension storage device; the lysis solution supply module comprises a lysis solution supply pipeline 20, which is in communication with an outlet valve of a lysis solution storage device, and the lysis solution supply pipeline 20 is in communication with the cell suspension supply pipeline 10 at a first mixing point 40; in some embodiments, the device further comprises a first flow detection element 11 for detecting the cumulative flow of the cell suspension flowing through the cell suspension supply pipeline 10 and regulating the relevant process flow response; in some embodiments, the device further comprises a second flow detection element 21 for detecting the cumulative flow of the lysis solution flowing into the first mixing point 40 through the lysis solution supply pipeline 20 and regulating the relevant process flow response; since there is a proportional relationship between the cell suspension and the lysis solution, the cumulative flow of one can be used to calculate the cumulative flow of the other, so in some embodiments, only the first flow detection element 11 or only the second flow detection element 21 is provided, as preferred, such as the embodiment shown in Figure 1 The cell suspension supply pipeline 10 is provided with the first flow detection element 11, and the lysis solution supply pipeline 20 is provided with the second flow detection element 21.

[0067] The neutralization solution supply module comprises a neutralization solution supply pipeline 30, which is in communication with an outlet valve of a neutralization solution storage device, and the neutralization solution supply pipeline 30 is in communication with a mixed solution outlet pipeline 41 at a second mixing point 50; the device further comprises a third flow detection element 42 for detecting the cumulative flow of the preliminary mixed solution flowing through the mixed solution outlet pipeline 41 and regulating the relevant process flow response; specifically, such as the embodiment shown in Figure 1 The third flow detection element 42 is arranged on the mixed solution outlet pipeline 41. The control device based on the cell lysis process of the present application detects the supply of the cell suspension through the first flow detection element 11, detects the supply of the lysis solution through the second flow detection element 21, and detects the flow of the preliminary mixed solution formed by the cell suspension and the lysis solution through the third flow detection element 42, thereby regulating the flow of each liquid in the entire cell lysis process, so as to ensure that the mixing reaction between the cell suspension, the lysis solution and the neutralization solution meets the proportioning requirements, the cell suspension can be completely consumed, waste is avoided, and batch consistency in mass production is ensured.

[0068] The device in this embodiment of the invention further includes an inert replenishment fluid supply module 60, connected to the cell suspension supply line 10. The inert replenishment fluid supply module 60 supplies inert replenishment fluid to the cell suspension supply line 10, ensuring continuous flow of the cell suspension within the line. The inert replenishment fluid does not react with the cell suspension, lysis buffer, or neutralizing solution. Its function is not only to propel the cell suspension in the cell suspension supply line 10 towards the first mixing point 40, preventing cell suspension residue in the line, but also to prevent preliminary mixture residue in the first mixing point 40, thus avoiding loss and waste; subsequently, it promotes the mixing of the preliminary mixture with the neutralizing solution. Figure 2 As shown, the inert replenishment fluid supply module 60 also includes an inert replenishment fluid storage tank 63, which is placed on the ground beside the main body 70. Alternatively, in other embodiments, the inert replenishment fluid of the inert replenishment fluid supply module 60 may be stored in a storage bag or other storage device, which can be suspended or placed on other storage devices. The inert replenishment fluid can be water or other neutral liquids that do not react with cell suspensions, lysis buffers, and neutralization solutions.

[0069] To ensure that no cell suspension and preliminary mixture remain, the control device also includes an inert replenishment time control unit (not shown). The inert replenishment time control unit is communicatively connected to the inert replenishment supply module 60 and is used to control the supply time of the inert replenishment. By automatically controlling the supply time of the inert replenishment, it ensures that all residual cell suspension in the cell suspension supply line 10 is pushed into the first mixing point 40 to mix with the lysis solution, thereby promoting the mixing of the preliminary mixture with the neutralization solution.

[0070] The inert replenishment time control unit can be a timer. The preset time of the timer is set by the operator, and the specific supply time can be determined by long-term experiments, practical observation or simulation. The inert replenishment supply time of the inert replenishment supply module 60 can also be executed according to the preset time, which is suitable for mass production.

[0071] like Figure 1 In the illustrated embodiment, the inert replenishment fluid supply module 60 is also connected to the lysis buffer supply line 20 to supply inert replenishment fluid to the lysis buffer supply line 20 so that the lysis buffer in the lysis buffer supply line 20 flows continuously. The purpose of using inert replenishment fluid to drive the lysis buffer is, on the one hand, to reduce the amount of lysis buffer used and avoid lysis buffer residue in the lysis buffer supply line 20, and on the other hand, to balance the pH value of the initial mixture and prevent excessive lysis buffer from flowing into the first mixing point 40 and mixing with the cell suspension, thereby disrupting the set mixing ratio.

[0072] Of course, in other embodiments, such as Figure 4In the second embodiment shown, the inert replenishment fluid supply module 60 can be connected only to the cell suspension supply line 10 and not to the lysis buffer supply line 20. The lysis buffer supply line 20 can stop supplying the lysis buffer within a set time after the inert replenishment fluid supply module 60 starts supplying the inert replenishment fluid, so as to ensure that the cumulative flow of the lysis buffer meets the ratio requirements with the cell suspension.

[0073] Specifically, such as Figure 1 In the first embodiment shown, a fifth valve 61 is provided between the inert replenishment fluid supply module 60 and the cell suspension supply pipeline 10, and a sixth valve 62 is provided between the inert replenishment fluid supply module 60 and the lysis buffer supply pipeline 20. When the fifth valve 61 is opened, it connects the inert replenishment fluid supply module 60 and the cell suspension supply pipeline 10. When the sixth valve 62 is opened, it connects the inert replenishment fluid supply module 60 and the lysis buffer supply pipeline 20, thereby realizing automatic control of the supply of inert replenishment fluid. This helps to reduce errors such as human operation errors or low automation of the solution preparation system.

[0074] In some embodiments of this utility model, such as Figure 4 Example 2 and Figure 6 In Embodiment 4, the lysis buffer supply module is a single-line flow module, that is, it only includes the lysis buffer supply line 20 and its components. The neutralizing solution supply module is a single-line flow module, that is, it only includes the neutralizing solution supply line 30 and its components. In these embodiments, the time for the lysis buffer to flow to the first mixing point 40 can be controlled by controlling the release time and length of the lysis buffer supply line 20, so that the lysis buffer arrives at the first mixing point 40 before or simultaneously with the cell suspension. The time for the lysis buffer to flow to the second mixing point 50 can be controlled by controlling the release time and length of the neutralizing solution supply line 30, so that the neutralizing solution arrives at the second mixing point 50 before or simultaneously with the preliminary mixture.

[0075] In some preferred embodiments, such as Figure 1 Example 1 and Figure 5In the third embodiment of the present application, the lysis solution supply module further comprises a lysis solution return circuit 22 in communication with the lysis solution supply circuit 20 to form a lysis solution self-circulation circuit, so that the lysis solution can be released in advance to fill the lysis solution self-circulation circuit; the connection between the lysis solution supply circuit 20 and the lysis solution return circuit 22 defines a lysis solution outlet point 28, which is closer to the first mixing point 40 than the starting end of the lysis solution supply circuit 20, has a shorter route, and the time for the lysis solution to reach the first mixing point 40 is more controllable; the distance between the first flow detection element 11 and the first mixing point 40 is D1, the distance between the lysis solution outlet point 28 and the first mixing point 40 is D2, and the distance between the starting end of the cell suspension supply circuit 10 and the first mixing point 40 is D3, which satisfy D2≤D1<D3; the lysis solution supply circuit 20 is used to close the lysis solution return circuit 22 and open the lysis solution supply circuit 20 when the cumulative flow of the cell suspension detected by the first flow detection element 11 is greater than 0, so that the lysis solution in the lysis solution supply circuit 20 flows to the first mixing point 40, and the cell suspension in the cell suspension supply circuit 10 flows to the first mixing point 40 faster, avoiding the loss of cell suspension empty flow, and also taking into account that the lysis solution and the cell suspension basically reach the first mixing point 40 at the same time or the lysis solution reaches the first mixing point 40 slightly earlier than the cell suspension under different flow rate ratios, reducing material waste. The second flow detection element 21 is arranged downstream of the lysis solution outlet point 28, which can accurately detect the cumulative flow of the lysis solution reacted with the cell suspension, and ensure that the mixing reaction between the cell suspension and the lysis solution and the neutralizing solution meets the ratio requirements.

[0076] As Figure 1 and Figure 5As shown, the neutralizing liquid supply module further comprises a neutralizing liquid return circuit 31 in communication with the neutralizing liquid supply pipeline 30 to form a neutralizing liquid self-circulation pipeline, so that the neutralizing liquid can be released early and fill the neutralizing liquid self-circulation pipeline; the connection between the neutralizing liquid supply pipeline 30 and the neutralizing liquid return circuit 31 defines a neutralizing liquid outlet point 37, which is closer to the second mixing point 50 relative to the starting end of the neutralizing liquid supply pipeline 30, has a shorter route, 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 D5≤D4<D6; the neutralizing liquid supply pipeline 30 is used to close the neutralizing liquid return 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 in the neutralizing liquid self-circulation pipeline can quickly respond, and is relatively faster than the preliminary mixed liquid in the mixed liquid outlet pipeline 41 to flow to the second mixing point 50, avoiding the loss of the preliminary mixed liquid due to empty flow, which cannot be fully mixed with the neutralizing liquid, ensuring that the mixing reaction between the cell suspension, the lysis liquid and the neutralizing liquid meets the proportioning requirements; it can also take into account that under different flow rate proportions, 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, reducing material waste.

[0077] The device main body 70 is provided with a control module, which can include a combination logic controller or a microprogram controller, mainly used for accepting signals and sending instructions, and the specific execution can be realized by hardware such as a control circuit. The control module is generally provided with a timer or a clock module to unify the time of the entire control device.

[0078] The cell suspension supply pipeline 10 is provided with a first supply pump 12, and the control module is in communication connection with the first supply pump 12 for controlling the operation of the first supply pump 12. The first supply pump 12 is automatically controlled to be turned on to push the cell suspension to flow in the cell suspension supply pipeline 10 to the first mixing point 40. The first supply pump 12 is a peristaltic pump, which is in wired or wireless communication connection with the control module. The control mode of the control module for the first supply pump 12 can be a conventional on-off circuit, which controls the start and stop of the first supply pump 12 through a logic level. The specific control mode is a conventional technology, which will not be described here. The control module is arranged in the device main body 70, which is closer to the first supply pump 12 and has a stronger signal, which is more conducive to the automatic control of the first supply pump 12.

[0079] The lysis solution supply pipeline 20 is provided with a second supply pump 23 and a first valve 24, the first valve 24 is located between the lysis solution outlet point 28 and the first mixing point 40, the lysis solution loop 22 is provided with a second valve 25, and the control module is in communication connection with the second supply pump 23, the first valve 24, the second valve 25 and the first flow detection element 11 respectively, and is used for controlling the work of the second supply pump 23. Similarly, the control module controls the second supply pump 23 in a conventional switching circuit mode, and controls the start and stop of the second supply pump 23 through a logic level. The specific control mode is a conventional technology, and will not be described here. The control module is arranged in the equipment main body 70, and is closer to the second supply pump 23, the first valve 24, the second valve 25 and the first flow detection element 11, the signal is stronger, the signal transmission speed is faster, and the precise and timely automatic control is beneficial.

[0080] The start time of the second supply pump 23 and the second valve 25 can be set by an operator and recorded in the control module, and the second supply pump 23 and the second valve 25 are automatically started at the time. The start time of the first valve 24 is determined by the first flow detection element 11. When the cumulative flow of the cell suspension detected by the first flow detection element 11 is greater than 0, the first flow detection element 11 sends a signal to the control module, and the signal can be a logic level. After the control module receives the logic level, the control module controls the first valve 24 to open and controls the second valve 25 to close, so that the lysis solution flows out of the lysis solution outlet point 28 and flows to the first mixing point 40.

[0081] The second supply pump 23 is located upstream of the lysis solution outlet point 28. 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 prepares for the lysis solution to flow quickly from the lysis solution outlet point 28 to the first mixing point 40. Then, after the first flow detection element 11 detects the cell suspension, it means that 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 flow out of the lysis solution outlet point 28 and quickly flow to the first mixing point 40, so that the cell suspension cannot be mixed with the lysis solution, causing the loss of the cell suspension.

[0082] The second supply pump 23 is a peristaltic pump, the first valve 24 and the second valve 25 are electromagnetic valves, and are in wired communication connection or wireless communication connection with the control module.

[0083] The neutralizing liquid supply pipeline 30 is provided with a third supply pump 32 and a third valve 33, the third valve 33 is located between the neutralizing liquid outlet point 37 and the second mixing point 50, the neutralizing liquid return pipeline 31 is provided with a fourth valve 34, and the control module is in communication connection 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 for controlling the work of the third supply pump 32. Similarly, the control module controls the third supply pump 32 in a conventional switching circuit mode, and controls the start and stop of the third supply pump 32 through a logic level. The specific control mode is a conventional technology, and will not be described here. The control module is arranged in the equipment main body 70, and is closer to the third supply pump 32, the third valve 33, the fourth valve 34 and the third flow detection element 42, the signal is stronger, the signal transmission speed is faster, and the precise and timely automatic control is beneficial.

[0084] The start time of the third supply pump 32 and the fourth valve 34 can be set by an operator and recorded in the control module, and the third valve 33 is started automatically at the time. When the third flow detection element 42 detects that the cumulative flow of the preliminary mixed liquid is greater than 0, the third flow detection element 42 sends a signal to the control module, and the signal can be a logic level. After the control module receives the logic level, the third valve 33 is controlled to be opened, and the fourth valve 34 is controlled to be closed, so that the neutralizing liquid flows out of the neutralizing liquid outlet point 37 and flows to the second mixing point 50.

[0085] The third supply pump 32 is located upstream of the neutralizing liquid outlet point 37, the control module preferentially opens the third supply pump 32 and the fourth valve 34, so that the neutralizing liquid fills the neutralizing liquid self-circulation pipeline, and prepares for rapid flow from the neutralizing liquid outlet point 37 to the second mixing point 50. After the third flow detection element 42 detects the preliminary mixed liquid, the fourth valve 34 is closed and the third valve 33 is opened, so that the neutralizing liquid can flow out of the neutralizing liquid outlet point 37 and quickly flow to the second mixing point 50, avoiding that part of the preliminary mixed liquid cannot be mixed with the neutralizing liquid when the preliminary mixed liquid flows to the second mixing point 50 first, causing loss of the preliminary mixed liquid.

[0086] The third supply pump 32 is a peristaltic pump, the third valve 33 and the fourth valve 34 are solenoid valves, and are in wired communication connection or wireless communication connection with the control module.

[0087] As Figure 2 and Figure 3As shown in FIG. 1, a first mixing device is connected downstream of the first mixing point 40, 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 mixing of the cell suspension and the lysis solution, and the first mixing device is used for the thorough mixing of the cell suspension and the lysis solution. The first mixing device includes a first spiral mixer 43 and a coil mixer 44. The cell suspension and the lysis solution are thoroughly mixed in the first spiral mixer 43 and then thoroughly reacted in the coil mixer 44 to form a preliminary mixed solution.

[0088] As shown in FIG. 1, a first mixing device is connected downstream of the first mixing point 40, 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 mixing of the cell suspension and the lysis solution, and the first mixing device is used for the thorough mixing of the cell suspension and the lysis solution. The first mixing device includes a first spiral mixer 43 and a coil mixer 44. The cell suspension and the lysis solution are thoroughly mixed in the first spiral mixer 43 and then thoroughly reacted in the coil mixer 44 to form a preliminary mixed solution. Figure 2 and Figure 3 As shown in FIG. 1, a first mixing device is connected downstream of the first mixing point 40, 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 mixing of the cell suspension and the lysis solution, and the first mixing device is used for the thorough mixing of the cell suspension and the lysis solution. The first mixing device includes a first spiral mixer 43 and a coil mixer 44. The cell suspension and the lysis solution are thoroughly mixed in the first spiral mixer 43 and then thoroughly reacted in the coil mixer 44 to form a preliminary mixed solution.

[0089] 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 structure is not described here. The first spiral mixer 43, the coil mixer 44, and the second spiral mixer 51 can be fixed on the equipment body 70 or on a storage device near the equipment body 70. The storage device can be a shelf, a trolley, or other equipment, or a wall or a floor. Figure 7 As shown in FIG. 5, the storage device is a trolley, the first spiral mixer 43 and the second spiral mixer 51 are fixed on the trolley in an inclined manner, and the coil mixer 44 is placed on the trolley for easy movement. In some cases, the mixing reaction needs a long time, and the length of the pipe in the coil mixer 44 is relatively long, which can be placed on the upper and lower layers of the trolley, respectively. Of course, in other embodiments with a relatively short mixing reaction time, the coil mixer 44 is placed on one layer of the trolley, and the length of the pipe in the coil mixer 44 is set as needed, which is not described here. The other structures of the fifth embodiment are the same as those of the first embodiment.

[0090] The arrangement of the cell suspension supply module and the lysis solution supply module is different based on different cell suspensions.

[0091] As shown in FIG. 1, a first mixing device is connected downstream of the first mixing point 40, 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 mixing of the cell suspension and the lysis solution, and the first mixing device is used for the thorough mixing of the cell suspension and the lysis solution. The first mixing device includes a first spiral mixer 43 and a coil mixer 44. The cell suspension and the lysis solution are thoroughly mixed in the first spiral mixer 43 and then thoroughly reacted in the coil mixer 44 to form a preliminary mixed solution. Figure 1In the illustrated embodiment, the first flow detection element 11 is positioned upstream of the first supply pump 12 on the cell suspension supply line. Although the first flow detection element 11 is relatively close to the control module within the device body 70, it takes a certain amount of time for the first flow detection element 11 to detect the cell suspension and send a signal to cause the lysis buffer to flow out of the lysis buffer outlet 28. Positioning the first flow detection element 11 upstream of the first supply pump 12 ensures that the distance between the first flow detection element 11 and the first mixing point 40 is appropriate, preventing the cell suspension from flowing to the first mixing point 40 before the lysis buffer and causing cell suspension loss.

[0092] like Figure 1 In the illustrated embodiment, the cell suspension, due to the presence of solids such as cells, has a high solid content and high viscosity, resulting in a slow flow rate. The lysis buffer, on the other hand, has a lower fluid concentration and viscosity, allowing for more controllable flow rate. Therefore, the cell suspension supply module and the lysis buffer supply module are arranged longitudinally, with the cell suspension supply module above and the lysis buffer supply module below. The cell suspension supply pipeline 10 includes a first transverse flow section 13 connected to the first mixing point 40, ensuring smooth flow of the cell suspension within the cell suspension supply pipeline 10 under the influence of gravity. The impact is relatively small, which helps to ensure the stability and uniformity of the cell suspension feed performance and flow rate. The lysis buffer supply line 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. The first longitudinal flow section 27 connects the second transverse flow section 26 and the first mixing point 40 from bottom to top. The lysis buffer flows smoothly in the second transverse flow section 26 and flows upward in the first longitudinal flow section 27. Since the adhesion between the lysis buffer and the inner wall of the pipe is small, it is easier to flow upward and control the flow rate.

[0093] like Figure 1 As shown, the second supply pump 23 on the lysis buffer supply line 20 is located on the second transverse flow section 26, and the second flow detection element 21 is located downstream of the second supply pump 23, on the first longitudinal flow section 27. The placement of the second supply pump 23 on the second transverse flow section 26 facilitates the smooth flow of the lysis buffer in the lysis buffer supply line 20. The function of the second flow detection element 21 is to detect the cumulative flow rate at the lysis buffer outlet point 28. The first longitudinal flow section 27 is closer to the first mixing point 40, ensuring the accuracy of the second flow detection element 21 in detecting the cumulative flow rate of the lysis buffer. This ensures that the mixing reaction between the cell suspension, lysis buffer, and neutralizing solution meets the required ratio, that the cell suspension is completely consumed without waste, and that batch-to-batch consistency is maintained during mass production.

[0094] like Figure 2 and Figure 3As shown, the main body 70 of the device includes four vertically arranged sides, namely the first side 71, the second side 72, the third side 73, and the fourth side 74. Part of the cell suspension supply pipeline 10 is located on the first side 71, and the first flow detection element 11 is located on the first side 71. Another part of the cell suspension supply pipeline 10 is located on the second side 72, and the first supply pump 12 is located on the second side 72. Part of the lysis buffer supply pipeline 20 is located on the first side 71, and part of the lysis buffer supply pipeline 20 is located on the second side 72. The second supply pump 23 and the second flow detection element 21 are located on the second side 72. By placing the cell suspension supply pipeline 10 and the lysis buffer supply pipeline 20 on the sides of the main body 70, the installation space on the sides of the main body 70 is fully utilized, and the components on each supply pipeline are closer to the control module, which is beneficial for signal transmission and overall compact design. The cell suspension supply pipeline 10 and the lysis buffer supply pipeline 20 are more closely related, so they are both located on the first side 71 and the second side 72 for easy manual observation.

[0095] In some embodiments, the cell suspension has a lower fluid concentration and viscosity, or is close to the fluid concentration and viscosity of the lysis buffer, which allows for... Figure 6 In the embodiment shown, the cell suspension supply module and the lysis buffer supply module are arranged longitudinally, with the cell suspension supply module at the bottom and the lysis buffer supply module at the bottom. The cell suspension supply line 10 includes a first transverse flow section 13 and a third longitudinal flow section 14, which is connected from bottom to top to the first mixing point 40. The lysis buffer supply line 20 includes a second transverse flow section 26 and a first longitudinal flow section 27, which is connected from top to bottom to the first mixing point 40.

[0096] exist Figure 1 In the illustrated embodiment, the first supply pump 12 on the cell suspension supply line 10 is disposed on the first transverse flow section 13, and the first flow detection element 11 is located upstream of the first supply pump 12. The first flow detection element 11 needs a certain amount of time from detecting the cell suspension to sending a signal to cause the lysis buffer to flow out of the lysis buffer outlet point 28. By placing the first flow detection element 11 upstream of the first supply pump 12, the lysis buffer receives the signal of the presence of the cell suspension earlier and is released in time and flows to the first mixing point 40, thus avoiding the loss of cell suspension due to the cell suspension flowing to the first mixing point 40 before the lysis buffer.

[0097] In other embodiments, such as Figure 5As shown, the first supply pump 12 on the cell suspension supply line 10 is set on the first transverse flow section 13. 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 rate of the cell suspension is relatively slow, allowing sufficient communication time and feedback time. This can still prevent the cell suspension from flowing to the first mixing point 40 before the lysis solution, thus avoiding cell suspension loss.

[0098] Generally, the initial mixture contains solids such as cell debris and biological products, resulting in a higher solids content and viscosity than the neutralization solution. Therefore, if... Figure 1 In the illustrated embodiment, the liquid outlet pipe 41 of the first mixing point 40 and the second mixing point 50 are located above the neutralizing liquid supply module. The initial mixture can be facilitated by gravity to flow more smoothly from the initial mixture outlet pipe 41 to the second mixing point 50. The neutralizing liquid supply pipe 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 neutralizing liquid has little adhesion to the inner wall of the pipe, it is easier to flow upward and control the flow rate.

[0099] like Figure 2 and Figure 3 As shown, part of the neutralizing liquid supply pipeline 30 is located on the third side 73, and the third supply pump 32 and the third flow detection element 42 are also located on the third side 73. This makes full use of the installation space on the side of the main body of the equipment. The neutralizing liquid supply pipeline 30 and the mixed liquid outlet pipeline 41 are located on the same side, which strengthens the correlation and facilitates manual observation, allowing for timely detection of automatic control faults or other problems.

[0100] like Figure 3 As shown, the portion of the mixed liquid outlet pipeline 41 between the third flow detection element 42 and the second mixing point 50 is defined as the additional pipeline 46. A suspension bracket 75 is provided on the third side 73, and the additional pipeline 46 is rolled up and sleeved on the suspension bracket 75.

[0101] Different types of cell suspensions form preliminary mixtures with varying fluid concentrations and viscosities, resulting in different flow rates of the preliminary mixture in the mixture outlet pipeline. The path of the preliminary mixture from the third flow detection element 42 to the second mixing point 50 also needs to be adjusted to ensure that the neutralizing solution and the preliminary mixture reach the second mixing point 50 as simultaneously as possible. Therefore, an additional pipeline 46 is adopted to facilitate users in adjusting the distance from the third flow detection element 42 to the second mixing point 50 based on actual needs. At the same time, the additional pipeline 46, which is suspended in a roll using the suspension bracket 75, makes full use of the space on the main body of the equipment 70, resulting in a clearer and neater pipeline setup.

[0102] Corresponding to Figure 1 The control method of one embodiment of the present application is as follows:

[0103] Step one: preset relevant parameters according to the formula: the pump speed v1 of the first supply pump 12, the pump speed v2 of the second supply pump 23, and the pump speed v3 of the third supply pump 32, the first flow setting value Q1, and Q1 is less than the processing volume of the cell suspension, the second flow setting value Q2, Q2 and Q1 meet the ratio requirement, and the preset time T;

[0104] Step two: open the second valve 25, the second supply pump 23 and the fourth valve 34, the third supply pump 32, keep the first valve 24 and the third valve 33 closed, the lysate flows in the lysate self circulation pipeline, and the neutralizing liquid flows in the neutralizing liquid self circulation pipeline;

[0105] Step three: open the first supply pump 12 to start supplying the cell suspension;

[0106] Step four: when the cumulative flow 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, opens the first valve 24 to make the lysate flow out of the lysate outlet point 28, and the cumulative flow of the lysate detected by the second flow detection element 21 is greater than 0, the control module records the time point as the second time, 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 reacted in the coil mixer 44 to form the preliminary mixed liquid;

[0107] Step five: when the cumulative flow 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, the neutralizing liquid flows out of the neutralizing liquid outlet point 37, flows to the second mixing point 50, the preliminary mixed liquid and the neutralizing liquid meet at the second mixing point 50, and are fully mixed and reacted in the second spiral mixer to form the final mixed liquid, which is finally input into the collection bag 53, as shown in Figure 2 or Figure 7 ;

[0108] Step six: when the cumulative flow of the lysate detected by the second flow detection element 21 reaches the second flow setting value, open the fifth valve 61 and the sixth valve 62, the inert supplement liquid supply module 60 supplies inert supplement liquid to the cell suspension supply pipeline 10 to make the cell suspension in the cell suspension supply pipeline 10 flow continuously, and supplies inert supplement liquid to the lysate supply pipeline 20 to make the lysate in the lysate supply pipeline 20 flow continuously;

[0109] It should be noted that step six does not necessarily start after step five, but can be earlier than step five, or can be performed simultaneously with step five.

[0110] Step seven: time counting for the supply time of the inert replenishment liquid supply module 60 until any one of the difference between the third time and the first time or the difference between the third time and the second time or the preset time T is reached, the first supply pump 12, the second supply pump 23 and the third supply pump 32 are stopped, and the cell lysis reaction is completed.

[0111] The time in the above steps is recorded by a clock module in the control module, for example, the time of step two is 8:00 of the system time, the time of step three is 8:10 of the system time, the first time is 8:15 of the system time, the second time is 8:16 of the system time, the third time is 9:00 of the system time, and 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 replenishment liquid supply module 60 is stopped when the supply time accumulates for 44 minutes, and at the same time, the first supply pump 12, the second supply pump 23 and the third supply pump 32 are stopped, realizing the high automation of the whole system, avoiding the personnel operation error or the low intelligent automatic control of the liquid preparation system, and thus ensuring the batch consistency in the mass production process.

[0112] The above embodiment is only a preferred embodiment of the present application, and cannot be used to limit the scope of protection of the present application. Any non-substantial change and replacement made by a person skilled in the art on the basis of the present application shall belong to the scope of protection required by the present application.

Claims

1. A control device based on a cell lysis process, comprising a device body, a cell suspension supply module, a lysis solution supply module, a neutralization solution supply module, and a first mixing point and a second mixing point, a mixed solution outlet pipeline being connected between the first mixing point and the second mixing point. Characterized in that, the cell suspension supply module comprises a cell suspension supply pipeline, the lysis solution supply module comprises a lysis solution supply pipeline, and the lysis solution supply pipeline and the cell suspension supply pipeline are communicated at the first mixing point; the neutralization solution supply module comprises a neutralization solution supply pipeline, and the neutralization solution supply pipeline and the mixed solution outlet pipeline are communicated at the second mixing point; at least one of the cell suspension supply pipeline, the lysis solution supply pipeline, and the neutralization solution supply pipeline is fixedly connected to the device body; further comprising a first flow detection element, a second flow detection element, and a third flow detection element, the first flow detection element being arranged on the cell suspension supply pipeline, the second flow detection element being arranged on the lysis solution supply pipeline, and the third flow detection element being arranged on the mixed solution outlet pipeline; further comprising an inert supplement solution supply module connected to the cell suspension supply pipeline, the inert supplement solution supply module being used to supply inert supplement solution to the cell suspension supply pipeline to continuously flow the cell suspension in the cell suspension supply pipeline.

2. The control device of claim 1, wherein further comprising an inert supplement solution time self-control unit, the inert supplement solution time self-control unit being communicatively connected to the inert supplement solution supply module and being used to control the supply time of the inert supplement solution.

3. The control device of claim 1, characterized in that, the lysis solution supply module further comprises a lysis solution return pipeline forming a lysis solution self-circulation pipeline in communication with the lysis solution supply pipeline, a connection position of the lysis solution supply pipeline and the lysis solution return pipeline is defined as a lysis solution outlet point, a position of the second flow detection element on the lysis solution supply pipeline is downstream of the lysis solution outlet point, a distance from the first flow detection element to the first mixing point is D1, a distance from the lysis solution outlet point to the first mixing point is D2, and a distance from a starting end of the cell suspension supply pipeline to the first mixing point is D3, satisfying D2≤D1<D3; and / or the neutralization solution supply module further comprises a neutralization solution return pipeline forming a neutralization solution self-circulation pipeline in communication with the neutralization solution supply pipeline, a connection position of the neutralization solution supply pipeline and the neutralization solution return pipeline is defined as a neutralization solution outlet point, a distance from the third flow detection element to the second mixing point is D4, a distance from the neutralization solution outlet point to the second mixing point is D5, and a distance from a starting end of the mixed solution outlet pipeline to the second mixing point is D6, satisfying D5≤D4<D6.

4. The control device of claim 3, wherein a control module is arranged in the device body, a first supply pump is arranged on the cell suspension supply pipeline, the control module is communicatively connected to the first supply pump, and / or The device body is provided with a control module, the lysis liquid supply pipeline is provided with a second supply pump and a first valve, the position of the second supply pump on the lysis liquid supply pipeline is upstream of the lysis liquid outlet point, the first valve is located between the lysis liquid outlet point and the first mixing point, the lysis liquid circuit is provided with a second valve, and the control module is in communication connection with the second supply pump, the first valve, the second valve and the first flow detection element respectively; And / or The device body is provided with a control module, the neutralization liquid supply pipeline is provided with a third supply pump and a third valve, the position of the third supply pump on the neutralization liquid supply pipeline is upstream of the neutralization liquid outlet point, the third valve is located between the neutralization liquid outlet point and the second mixing point, the neutralization liquid circuit is provided with a fourth valve, and the control module is in communication connection with the third supply pump, the third valve, the fourth valve and the third flow detection element respectively.

5. The control device of claim 4, wherein The position of the first flow detection element on the cell suspension supply pipeline is upstream of the first supply pump.

6. The control device of claim 4, wherein The cell suspension supply module and the lysis liquid supply module are arranged in a longitudinal direction, the cell suspension supply pipeline comprises a first transverse flow section connected with the first mixing point, the lysis liquid supply pipeline comprises a second transverse flow section and a first longitudinal flow section, the second transverse flow section is located below the first transverse flow section, and the first longitudinal flow section connects the second transverse flow section 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 neutralization liquid supply module, the neutralization liquid supply pipeline comprises a third transverse flow section and a second longitudinal flow section, and the second longitudinal flow section connects the third transverse flow section and the second mixing point from bottom to top.

7. The control device of claim 6, wherein The first supply pump on the cell suspension supply pipeline is arranged on the first transverse flow section; and / or The second supply pump on the lysis liquid supply pipeline is arranged on the second transverse flow section, and the second flow detection element is arranged on the first longitudinal flow section.

8. The control device of claim 4, wherein The device body comprises four vertically arranged side surfaces, which are divided into a first side surface, a second side surface, a third side surface and a fourth side surface; Part of the cell suspension supply pipeline is arranged on the first side surface, and the first flow detection element is arranged on the first side surface; part of the cell suspension supply pipeline is arranged on the second side surface, and the first supply pump is arranged on the second side surface; and / or Part of the lysis liquid supply pipeline is arranged on the first side surface, part of the lysis liquid supply pipeline is arranged on the second side surface, and the second supply pump and the second flow detection element are arranged on the second side surface; and / or Part of the neutralization liquid supply pipeline is arranged on the third side surface, and the third supply pump and the third flow detection element are arranged on the third side surface.

9. The control device of claim 8, wherein The part of the mixed liquid outlet pipeline between the third flow detection element and the second mixing point is defined as an additional pipeline, a hanging bracket is arranged on the third side surface, and the additional pipeline is sleeved on the hanging bracket in a roll.

10. The control device of claim 1, wherein, The inert supplement liquid supply module is also connected with the lysis liquid supply pipeline, and is used for supplying inert supplement liquid to the lysis liquid supply pipeline, so that the lysis liquid in the lysis liquid supply pipeline continuously flows. A fifth valve is arranged between the inert supplement liquid supply module and the cell suspension supply pipeline, and a sixth valve is arranged between the inert supplement liquid supply module and the lysis liquid supply pipeline.

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