WO2024041201A1

WO2024041201A1 - Flow electroporation apparatus and control method therefor

Info

Publication number: WO2024041201A1
Application number: PCT/CN2023/104683
Authority: WO
Inventors: 戴晓兵
Original assignee: 苏州壹达生物科技有限公司
Priority date: 2022-08-25
Filing date: 2023-06-30
Publication date: 2024-02-29
Also published as: CN117660703A

Abstract

Provided are a flow electroporation apparatus and a control method therefor. The control method comprises: starting a first electrode assembly at a first moment, forming an electric field in an electroporation chamber of the first electrode assembly, and subjecting a first cell group to electroporation treatment in the electroporation chamber of the first electrode assembly; and starting a second electrode assembly at a second moment, forming an electric field in an electroporation chamber of the second electrode assembly, and subjecting a second cell group to electroporation treatment in the electroporation chamber of the second electrode assembly. The first moment is not equal to the second moment; the first cell group is not subjected to electroporation treatment when flowing through the electroporation chamber of the second electrode assembly, and the second cell group is not subjected to electroporation treatment when flowing through the electroporation chamber of the first electrode assembly. Each of the electrode assemblies does not need to continuously work for a long time, so that the flow electroporation apparatus as a whole can perform stable and efficient electroporation treatment on more continuously flowing cell suspensions.

Description

A flow electroporation device and its control method

Technical field

The present invention relates to the field of biotechnology, and in particular, to a flow electroporation device and a control method thereof.

Background technique

Electroporation is a technology that uses an electric field to act on the cell membrane to create micropores in the cell membrane that can pass through for foreign molecules such as DNA, RNA, and proteins. These micropores are instantly generated under the action of the electric field and can change with the electric field. disappear and recover, thereby avoiding permanent damage to cells. Electroporation technology can be used to introduce DNA, RNA, proteins, sugars, dyes, virus particles and other foreign molecules into prokaryotic or eukaryotic cells, which is called electrotransfection; electrotransfection technology is used in antibody proteins. It is widely used in many high-end biomedical technology fields such as production, in vitro diagnostic (IVD) reagent raw material production, immune cell gene editing and gene modification, and is an important technology for intracellular material delivery.

There are many methods for electroporation of cells. Among them, flow electroporation technology can continuously electroporate a flowing suspension containing cells (hereinafter referred to as cell suspension), which is more efficient and is especially suitable for large volumes. Batch processing of cell suspensions. A common flow electroporation device mainly includes a pair of planar electrodes for generating an electric field, and an electroporation chamber located between the two planar electrodes. The cell suspension can receive electric shock while flowing through the chamber, resulting in electroporation. , so that foreign molecules in the cell suspension can enter the target cells.

In the existing technology, most flow electroporation devices have the problem of low cell processing efficiency (such as electrotransfection efficiency). When the volume of cell suspension to be processed is large and the electrodes need to work for a long time, the electrotransfection efficiency and The problem of cell viability decreasing with time is particularly obvious.

On the one hand, the area of each planar electrode should not be too large. When the cell suspension flow rate is constant, the length of the flat electrode (along the direction of liquid flow) will affect the time for liquid flow in the electroporation chamber. Increasing the electrode length will increase the time for bubbles to contact the electrode surface when they flow through the electrode area, increasing The probability of bubble retention. The width of the flat electrode will affect the uniformity of the flow rate of the cell suspension in the electroporation chamber. According to fluid mechanics, the flow rate in the middle area of the fluid is fast and the flow rate in the edge area is slow. The larger the electrode width, the more obvious the flow rate deviation, which in turn affects the cells. The number of electric shocks is consistent (not every cell receives the optimal number of pulses), so increasing the length and width of the electrode will reduce the electrotransfection efficiency and/or cell survival rate in different aspects. At the same time, the distance between the two electrodes cannot be too large, otherwise the voltage and power of the power supply will be significantly increased, the economic cost will be increased, and a large number of cells will die due to problems such as excessive voltage and severe electrode heating. Therefore, the electroporation chamber of a pair of planar electrodes with a small area and a small distance is also small, and the volume of cell suspension that it can process per unit time is limited.

On the other hand, each electrode has a limited service life, making it difficult to maintain continuous operation at high cell processing efficiency (such as electrotransfection efficiency) for a long time. During the electroporation process, the cell suspension is in contact with the electrode, and the electrolytic water reaction easily occurs on the electrode surface, generating bubbles. At the same time, as the electrode works for a long time, more and more foam clusters (bubbles and dead cells) will adhere to the electrode surface. mixture), because the foam mass is not conductive, the electric field intensity distribution in the electroporation chamber will change, destroying the uniformity and stability of the entire electric field, thereby affecting the cell electrotransfection efficiency; at the same time, the foam mass occupies the electroporation chamber The chamber space will change the flow rate of cell suspension, resulting in changes in the number and intensity of electric shocks received by cells flowing through the electrode area, resulting in changes in electroporation results, which in turn affects the electrotransfection efficiency of cells. As the electrode works for a long time, the temperature of the electrode surface increases. At the same time, the Joule effect of the current causes the temperature of the cell suspension to increase, causing a large number of cell death and reducing the cell survival rate.

In summary, existing flow electroporation devices and their control methods are still unable to perform continuous, efficient, and stable electroporation of large-volume cell suspensions.

Contents of the invention

The purpose of the present invention is to address the problems existing in the prior art and provide a flow electroporation device and a control method thereof that can perform continuous, efficient, and stable electroporation processing of large-volume cell suspensions.

In order to achieve the above object, the technical solution adopted by the present invention is:

A flow electroporation device control method for electroporating a flowing cell suspension, the cell suspension containing a plurality of cell groups arranged sequentially along its own transport direction, each of the cell groups containing A plurality of suspended cells. The plurality of cell groups at least include a first cell group and a second cell group. Multiple groups of electrode assemblies are sequentially provided along the transmission direction. Each group of electrode assemblies has an electrode assembly for the cells. In an electroporation chamber for suspension circulation, the plurality of electrode assemblies at least include a first electrode assembly and a second electrode assembly, the first electrode assembly and the second electrode assembly are connected in series, and the control method includes:

The first electrode assembly is turned on, an electric field is formed in the electroporation chamber of the first electrode assembly, and the first cell group is subjected to electroporation treatment in the electroporation chamber of the first electrode assembly. ;

The second electrode assembly is turned on, an electric field is formed in the electroporation chamber of the second electrode assembly, and the second cell group is subjected to electroporation treatment in the electroporation chamber of the second electrode assembly. ;

The first cell group is not subjected to electroporation treatment when flowing through the electroporation chamber of the second electrode assembly, and the second cell group is not subjected to electroporation treatment when flowing through the electroporation chamber of the first electrode assembly. were not subjected to electroporation during treatment.

In some embodiments, there is no electric field in the electroporation chamber of the second electrode assembly when the first cell population flows through the electroporation chamber of the second electrode assembly. In some embodiments, there is no electric field in the electroporation chamber of the first electrode assembly when the second cell population flows through the electroporation chamber of the first electrode assembly.

In some embodiments, the first electrode assembly is turned on at the first moment, and the second electrode assembly is turned on at the second moment; the control method further includes: turning off the first electrode assembly at the third moment, so The electric field in the electroporation chamber of the first electrode assembly disappears, and the third time is later than the first time; the second electrode assembly is turned off at the fourth time, and all the parts of the second electrode assembly are closed. The electric field in the electroporation chamber disappears, and the fourth time is later than the second time; the first time and the second time are not equal, and/or, the third time is the same as the third time. The four moments are not equal. Therefore, the first electrode assembly and the second electrode assembly work in different time periods, and can electroporate different cell groups.

In some implementations, the first time is earlier than the second time, and the third time is earlier than the fourth time. Therefore, the first electrode assembly that is opened first is closed first, and the second electrode assembly that is opened later is closed later. Compared with a single group of electrode assemblies, both groups of electrode assemblies connected in series can work for a relatively shorter time, avoiding long-term work that may cause malfunctions. Efficiency is reduced.

In some implementations, the time interval between the first time and the third time is equal to the time interval between the second time and the fourth time. Therefore, the working time of the first electrode assembly and the second electrode assembly is equal, which helps to ensure the consistency of the electroporation treatment effect and facilitates the control of the flow electroporation device.

In some embodiments, the cell suspension is transferred from the first electrode assembly to the second electrode assembly, and the second time is later than the third time. At this time, the first electrode assembly located upstream works first. In order to prevent some cells from being repeatedly shocked and increasing the probability of death, the second electrode assembly located downstream is turned on after the first electrode assembly stops working for a period of time.

In some embodiments, the cell suspension is transferred from the second electrode assembly to the first electrode assembly, and the second time is earlier than the third time. At this time, the first electrode assembly located downstream works first. In order to avoid the electrotransfection efficiency being reduced due to failure of some cells to receive electric shock, the second electrode assembly located upstream is turned on in advance before the first electrode assembly is turned off.

In some embodiments, the plurality of cell groups further include a third cell group, which is located between the first cell group and the second cell group along the transport direction, and the A third cell population is subjected to electroporation while flowing through the electroporation chamber of the first electrode assembly, and the third cell population is also subjected to electroporation while flowing through the electroporation chamber of the second electrode assembly. Subjected to electroporation. No matter which one of the first electrode assembly and the second electrode assembly is upstream, during the time period when the two are switching work, the third cell group located between the first cell group and the second cell group will be in both the front and rear groups of electrode assemblies. received electric shock, but the sum of the time that each cell in the third cell group was electroporated in the two sets of electrode assemblies was approximately the same as the time that each cell in the first cell group or the second cell group was electroporated. , thereby improving the continuity and consistency of electroporation processing.

In some embodiments, each cell in the first cell population is electroporated for an equal amount of time as each cell in the second cell population is electroporated. This helps improve the consistency of electroporation treatment effects across the entire cell suspension.

In some embodiments, the plurality of electrode assemblies further include a third electrode assembly, and the cell suspension flows through the first electrode assembly, the second electrode assembly and the third electrode assembly sequentially along the transmission direction. The electroporation chamber of the electrode assembly; the control method includes the following steps in sequence:

S1, turn on the first electrode assembly at the first moment; S2, turn off the first electrode assembly at the third moment, the electric field in the electroporation chamber of the first electrode assembly disappears, and the third The time is later than the first time; S3. Turn on the second electrode assembly at the second time, and the second time is later than the third time; S4. Turn off the second electrode assembly at the fourth time, The electric field in the electroporation chamber of the second electrode assembly disappears, and the fourth time is later than the second time; S5. Turn on the third electrode assembly at the fifth time, and the third electrode An electric field is formed in the electroporation chamber of the component, and the fifth moment is later than the fourth moment; S6. Close the third electrode component at the sixth moment, and the electroporation of the third electrode component The electric field in the chamber disappears, and the sixth time is later than the fifth time.

Three sets of electrode assemblies connected in series can further improve the working efficiency of the flow electroporation device. The three sets of electrode assemblies work sequentially from upstream to downstream, and can electroporate more cell groups continuously and efficiently.

In some embodiments, at the third moment, along the transport direction, the end of the electroporation chamber of the first electrode assembly and the end of the electroporation chamber of the second electrode assembly are The cell group between the starting ends is a first transition group; at the second moment, along the transmission direction, all of the first transition group flows through the electroporation chamber of the second electrode assembly at the end of the At four moments, along the transmission direction, the cell group located between the end of the electroporation chamber of the second electrode assembly and the starting end of the electroporation chamber of the third electrode assembly is The second transition group; at the fifth moment, along the transport direction, the second transition group all flows through the end of the electroporation chamber of the third electrode assembly. This ensures that each cell arranged along the transport direction can receive electroporation treatment for approximately the same time, further improving the consistency of the electroporation treatment effect.

In some embodiments, the plurality of electrode assemblies further include a third electrode assembly, and the cell suspension flows through the third electrode assembly, the second electrode assembly and the first electrode assembly along the transmission direction. The electroporation chamber of the electrode assembly; the control method includes the following steps in sequence:

S1. Turn on the first electrode assembly at the first moment; S2. Turn on the second electrode assembly at the second moment, which is later than the first moment; S3. Turn off the electrode assembly at the third moment. The first electrode assembly, the electric field in the electroporation chamber of the first electrode assembly disappears, and the third time is later than the second time; S4, turn on the third electrode assembly at the fifth time, An electric field is formed in the electroporation chamber of the third electrode assembly, and the fifth time is later than the third time; S5. Close the second electrode assembly at the fourth time, and the second electrode assembly The electric field in the electroporation chamber disappears, and the fourth moment is later than the fifth moment; S6. Turn off the third electrode assembly at the sixth moment, and the electroporation of the third electrode assembly The electric field in the chamber disappears, and the sixth time is later than the fourth time.

Three sets of electrode assemblies connected in series can further improve the working efficiency of the flow electroporation device. The three sets of electrode assemblies work sequentially from downstream to upstream, and can electroporate more cell groups continuously and efficiently.

In some embodiments, at the second moment, along the transport direction, the end of the electroporation chamber of the second electrode assembly is between the end of the electroporation chamber of the first electrode assembly. The cell group between the starting ends is the first transition group; at the third moment, along the transmission direction, the first transition group all flows through the electroporation chamber of the first electrode assembly at the fifth moment, along the transmission direction, between the end of the electroporation chamber of the third electrode assembly and the starting end of the electroporation chamber of the second electrode assembly The cell group in between is the second transition group; at the fourth moment, along the transport direction, all the second transition group flows through the end of the electroporation chamber of the second electrode assembly. This ensures that each cell arranged along the transport direction can receive electroporation treatment for approximately the same time, further improving the consistency of the electroporation treatment effect.

The present invention also provides a flow electroporation device for performing electroporation treatment on cell suspension. The flow electroporation device includes multiple groups of electrode assemblies, and each group of electrode assemblies has a structure for the cell suspension. An electroporation chamber with liquid flow, the plurality of electrode assemblies at least include a first electrode assembly and a second electrode assembly, the first electrode assembly and the second electrode assembly are connected in series, the flow electroporation device further including a control system, the control system is used to control the first electrode assembly and the second electrode assembly to be turned on at different times, and/or, the control system is used to control the first electrode assembly and the second electrode assembly to be turned on at different times. Closed at different times.

In some embodiments, the plurality of groups of electrode assemblies further include a third electrode assembly, the first electrode assembly, the second electrode assembly and the third electrode assembly are connected in series, and the second electrode assembly is located at the Between the first electrode assembly and the third electrode assembly, the control system is also used to control the opening and closing of the third electrode assembly.

The present invention also provides a flow electroporation device for performing electroporation treatment on cell suspension. The flow electroporation device includes:

Multiple sets of electrode assemblies, each of which has an electroporation chamber through which the cell suspension can flow;

A plurality of electrode assemblies are connected through the connecting pipeline. The connecting pipeline includes a plurality of branches connected in parallel, and each of the branches is connected to at least one group of the electrode assemblies.

In some embodiments, the plurality of groups of electrode assemblies at least include a first electrode assembly and a second electrode assembly connected in series to one of the branches. In some embodiments, the control system uses the aforementioned control method to control the flow electroporation device.

Due to the application of the above technical solutions, in the flow electroporation device control method provided by the present invention, at least some of the different cell groups are electroporated by different electrode assemblies respectively, so that each group of electrode assemblies does not need to work continuously for a long time, effectively avoiding ① As the electrode works for a long time, the surface temperature of the electrode increases. At the same time, the Joule effect of the current causes the temperature of the cell suspension to increase, causing a large number of cell death and a significant reduction in cell survival rate; ② As the electrode works for a long time , more and more foam clusters (a mixture of bubbles and dead cells) will adhere to the electrode surface, destroying the uniformity and stability of the electric field, resulting in a significant reduction in electrotransfection efficiency. The control method provided by the present invention enables the flow electroporation device as a whole to perform stable and efficient electroporation processing of a larger volume of continuously flowing cell suspension. In the flow electroporation device provided by the present invention, multiple groups of electrode assemblies are connected in series, and each electrode assembly can be opened and/or closed in a time-sharing manner through a control system. When the cell suspension to be processed is When the volume is constant, the working time of each set of electrode components is shortened, which helps to maintain the overall device's efficient and stable electrotransfection efficiency and cell survival rate for a longer period of time.

Description of drawings

In order to explain the technical solution of the present invention more clearly, the accompanying drawings needed to be used in the description of the embodiments will be briefly introduced below.

Figure 1 is a schematic diagram of the flow electroporation device in Embodiment 1 when it is in step S1; Figure 2 is a schematic diagram of the flow electroporation device in Embodiment 1 when it is in step S2; Figure 3 is a schematic diagram of the flow electroporation device in Embodiment 1 A schematic diagram of the flow electroporation device in step S3; Figure 4 is a schematic diagram of the flow electroporation device in step S4 in Example 1; Figure 5 is a schematic diagram of the flow electroporation device in step S5 in Example 1; Figure 6 is a schematic diagram of the implementation A schematic diagram of the flow electroporation device in step S6 in Example 1; Figure 7 is a schematic flow diagram of the control method of the flow electroporation device in Example 1; Figure 8 is a schematic diagram of the flow electroporation device in Step S1 in Example 2 ; Figure 9 is a schematic diagram of the flow electroporation device in Embodiment 2 when it is in step S2; Figure 10 is a schematic diagram of the flow electroporation device in Embodiment 2 when it is in step S3; Figure 11 is a schematic diagram of the flow electroporation device in Embodiment 2 The schematic diagram when the device is in step S4; Figure 12 is the schematic diagram when the flow electroporation device in Embodiment 2 is in step S5; Figure 13 is the schematic diagram when the flow electroporation device in Embodiment 2 is in step S6; Figure 14 is A schematic flow diagram of the flow electroporation device control method in Embodiment 2; Figure 15 is a schematic diagram of the flow electroporation device in Embodiment 3; Figure 16 is a schematic diagram of the flow electroporation device in Embodiment 4;

Among them: 1. Electrode assembly; 11. First electrode assembly; 12. Second electrode assembly; 13. Third electrode assembly; 2. Electroporation chamber; 21. Electrode inlet; 22. Electrode outlet; 3. Connecting pipeline ; 31. Liquid inlet pipe; 32. Liquid outlet pipe; 33. First connecting pipe; 34. Second connecting pipe; 35. First pipe group; 351. First main road; 352. First branch road; 36. The second pipe group; 361, the second main road; 362, the second branch road.

Detailed ways

The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art.

Example 1

Referring to FIGS. 1 to 7 , this embodiment provides a flow electroporation device and a control method thereof for electroporating cell suspension. The cell suspension to be processed contains multiple cell groups arranged sequentially along its own transmission direction (indicated by arrows in the figure, referred to as the transmission direction in this article). Each cell group contains multiple suspended cells. Including at least the first cell group and the second cell group. It should be noted that the definition of cell groups in the present invention is mainly to facilitate the explanation of the technical concept and working principle of the present invention, and does not mean that different cell groups are clearly separated and independent of each other. In fact, the continuously flowing cell suspension contains many suspended cells. These cells are roughly arranged in an orderly manner along the transmission direction. The structural properties of each cell are basically the same. In this embodiment, different cells are artificially distributed along the transmission direction. Cell groups are defined as multiple "cell groups", each of which contains varying numbers of cells. There are no clear boundaries between different cell groups, and they may even mix with each other in a small adjacent area.

In this embodiment, along the transmission direction, the first cell group is located downstream of the second cell group, and there is a third cell group between the first cell group and the second cell group. The third cell group occupies 30% of the cell suspension. The volume and the number of cells contained therein are significantly smaller than the volume of the cell suspension occupied by the first cell group and the second cell group and the number of cells contained therein. Different graphical symbols are used to represent different cell groups in each drawing of this embodiment, and illustrations are used to illustrate. These symbols are only abstract representations and have nothing to do with the actual shape and number of cells. They are intended to facilitate those skilled in the art to understand this embodiment. The dynamic change process of each step of the control method in the example.

Referring to Figure 1, in this embodiment, the flow electroporation device includes a connecting pipeline 3 and multiple sets of electrode assemblies 1. The multiple sets of electrode assemblies 1 are arranged sequentially along the transmission direction, and each set of electrode assemblies 1 has a space for cell suspension. Electroporation chamber 2 for fluid flow. In this embodiment, the bottom end and top end of each electroporation chamber 2 are respectively provided with an electrode inlet 21 and an electrode outlet 22. When the cell suspension flows through each electroporation chamber 2, it passes through the electrode inlet 21. flows to the corresponding electrode outlet 22.

As shown in FIG. 1 , the electrode assembly 1 at least includes a first electrode assembly 11 and a second electrode assembly 12 . The first electrode assembly 11 and the second electrode assembly 12 are connected in series. The electrode outlet 22 of the first electrode assembly 11 is connected to the second electrode assembly 12 . The electrode inlets 21 of the components 12 are connected through the connecting pipeline 3 , and the cell suspension is transferred from the first electrode component 11 to the second electrode component 12 . In this embodiment, the electrode assembly 1 specifically has three groups. In addition to the above-mentioned first electrode assembly 11 and the second electrode assembly 12, it further includes a third electrode assembly 13. The second electrode assembly 12 is disposed between the first electrode assembly 11 and the third electrode assembly 13. Among the electrode assemblies 13 , the first electrode assembly 11 , the second electrode assembly 12 and the third electrode assembly 13 are connected in series. Specifically, the connecting pipe 3 includes a liquid inlet pipe 31, a liquid outlet pipe 32, a first connecting pipe 33, a second connecting pipe 34, and so on. The connecting pipe 3 can be used to connect the electrode assemblies 1 , that is, multiple electrode assemblies 1 are connected through the connecting pipe 3 . Among them, the liquid inlet pipe 31 is connected with the electrode inlet 21 of the first electrode assembly 11, the first connecting pipe 33 is connected between the electrode outlet 22 of the first electrode assembly 11 and the electrode inlet 21 of the second electrode assembly 12, and the second connection pipe 33 is connected with the electrode inlet 21 of the first electrode assembly 11. The tube 34 is connected between the electrode outlet 22 of the second electrode assembly 12 and the electrode inlet 21 of the third electrode assembly 13 , and the liquid outlet pipe 32 is connected with the electrode outlet 22 of the third electrode assembly 13 . In this way, the cell suspension to be treated can be continuously input from the sampling container (not shown in the figure) through the liquid inlet pipe 31, and sequentially flow through the first electrode assembly 11, the second electrode assembly 12 and the third electrode assembly along the transmission direction. Electroporation chamber of 13 In chamber 2, the cell groups contained in the cell suspension are subjected to electroporation treatment in the electroporation chamber 2 of a certain group or two groups of electrode assemblies 1, and then flow out from the outlet pipe 32, and are finally collected into a collection container (Fig. (not shown) collected.

In this embodiment, the flow electroporation device also includes a control system (not shown in the figure). The control system is used to control the first electrode assembly 11 and the second electrode assembly 12 to be turned on at different times, and/or the control system uses The first electrode assembly 11 and the second electrode assembly 12 are controlled to be closed at different times. The control system specifically uses the control method provided in this embodiment to control the flow electroporation device, and can independently control the opening and closing of the first electrode assembly 11, the second electrode assembly 12, and the third electrode assembly 13.

In this embodiment, the control system adopts a time-sharing control method for each electrode assembly 1 to improve the electroporation processing efficiency and processing capacity of the entire flow electroporation device. In this embodiment, the so-called time-sharing control means that different electrode assemblies 1 do not work completely at the same time. The first electrode assembly 11, the second electrode assembly 12, and the third electrode assembly 13 are respectively turned on and/or turned off at different times. The groups of electrode assemblies 1 take over and work in turns over time. Through pre-planning and design, the opening and closing time of each group of electrode assemblies 1 is controlled, thereby effectively shortening the energized working time of each group of electrode assemblies 1 and avoiding the need for a single group of electrode assemblies 1 Problems such as local heating and reduced cell processing efficiency caused by long-term work, while maintaining the entire flow electroporation device can run continuously for a longer period of time with more stable electrotransfection efficiency, especially suitable for large-volume cell suspensions Continuous processing.

It should be noted that when a certain electrode assembly 1 is "turned on", "operated", etc. in this article, it means that an electric field is formed in the electroporation chamber 2 of the electrode assembly 1 and can affect the flow through the electroporation chamber at this time. The cell group in chamber 2 is electroporated; and when an electrode assembly 1 is "off", it means that the electrode assembly 1 is no longer energized, and the electric field in the electroporation chamber 2 disappears. At this time, the flow through the electroporation chamber The cell population in chamber 2 will not be subjected to electroporation. The electric field referred to herein can be a continuous electric field or a pulsed electric field. In this embodiment, it is specifically a pulsed electric field.

In this embodiment, the control method includes: turning on the first electrode assembly 11 at the first moment. At this time, an electric field is formed in the electroporation chamber 2 of the first electrode assembly 11, and the first cell group is exposed to the electric field of the first electrode assembly 11. The electroporation process is performed in the perforation chamber 2 (see Figures 1 to 2); at the second moment, the second electrode assembly 12 is turned on, an electric field is formed in the electroporation chamber 2 of the second electrode assembly 12, and the second cell group is The second electrode assembly 12 is subjected to electroporation treatment in the electroporation chamber 2 (see FIGS. 3 to 4 ); wherein the first time and the second time are not equal. The above-mentioned first cell group is not subjected to electroporation treatment when flowing through the electroporation chamber 2 of the second electrode assembly 12, and the second cell group is not subjected to electroporation treatment when flowing through the electroporation chamber 2 of the first electrode assembly 11. Electroporation treatment. In this embodiment, the third cell group is subjected to electroporation treatment when flowing through the electroporation chamber 2 of the first electrode assembly 11 , and when the third cell group flows through the electroporation chamber 2 of the second electrode assembly 12 Also subjected to electroporation.

In this embodiment, the control method also includes: closing the first electrode assembly 11 at a third moment, and the electric field in the electroporation chamber 2 of the first electrode assembly 11 disappears, and the third moment is later than the first moment (see Figure 2) ; At the fourth moment, the second electrode assembly 12 is closed, the electric field in the electroporation chamber 2 of the second electrode assembly 12 disappears, and the fourth moment is later than the second moment (see Figure 4); the third moment is different from the fourth moment. Equivalent.

In this embodiment, the first time is earlier than the second time, and the third time is earlier than the fourth time. Further, the time interval between the first moment and the third moment is equal to the time interval between the second moment and the fourth moment. The second moment is later than the third moment. That is to say, the second electrode assembly 12 is turned on after a period of time after the first electrode assembly 11 is turned off.

Further, the time for each cell in the first cell group to be subjected to electroporation treatment is equal to the time for each cell in the second cell group to be subjected to electroporation treatment. Specifically in this embodiment, the length and cross-sectional area of the electroporation chamber 2 of the first electrode assembly 11 and the second electrode assembly 12 are equal, and the cell suspension flows at a substantially constant flow rate, so that the first cell group Each cell in the second cell population was electroporated for approximately the same amount of time. Furthermore, the size of the electroporation chamber 2 of the third electrode assembly 13 is also equal to that of the first electrode assembly 11 and the second electrode assembly 12. Combined with an accurate time-sharing control method, it is possible to ensure that all the cells in the cell suspension are The cells are all subjected to electroporation treatment for the same time, further improving the consistency of the electroporation treatment effect. In this embodiment, the preset time for each cell to undergo electroporation treatment is defined as T.

Referring to Figures 1 to 7, each step of the control method of the flow electroporation device in this embodiment will be described in detail below. In this control method, all the cell groups in the cell suspension are named sequentially along the transmission direction as the first cell group, the third cell group, the second cell group, the fifth cell group and the fourth cell group, where the first cell group The fourth cell group is located at the most downstream and enters the liquid inlet pipe 31 first. The fourth cell group is located at the most upstream and enters the liquid inlet pipe 31 last.

The control method includes the following steps in sequence:

S1, open the first electrode assembly 11 at the first moment, see Figure 1; S2, close the first electrode assembly 11 at the third moment, the electric field in the electroporation chamber 2 of the first electrode assembly 11 disappears, the third moment is later At the first moment, see Figure 2; S3, the second electrode assembly 12 is turned on at the second moment, which is later than the third moment, see Figure 3; S4, the second electrode assembly 12 is turned off at the fourth moment, the second The electricity of the electrode assembly 12 The electric field in the perforation chamber 2 disappears, and the fourth moment is later than the second moment, see Figure 4; S5, the third electrode assembly 13 is turned on at the fifth moment, and an electric field is formed in the electroporation chamber 2 of the third electrode assembly 13 , the fifth moment is later than the fourth moment, see Figure 5; S6, the third electrode assembly 13 is closed at the sixth moment, the electric field in the electroporation chamber 2 of the third electrode assembly 13 disappears, the sixth moment is later than the fifth moment time, see Figure 6.

That is to say, in this embodiment, along the transmission direction, the first electrode assembly 11 through which the cell suspension flows first is turned on first, then the second electrode assembly 12 in the middle is turned on, and the third electrode assembly 13 through which the cell suspension flows last is turned on. Turn on. Furthermore, after the previous group of electrode assemblies 1 is turned off and a period of time elapses, the latter group of electrode assemblies 1 is turned on again.

Referring to Figure 1, in this embodiment, the first moment is the moment when the first cell group has just entered or is about to enter the starting end of the electroporation chamber 2 of the first electrode assembly 11. Therefore, the liquid is fed from the liquid inlet pipe 31 to the first cell group. There is a brief delay between moments. Of course, due to actual operational errors, the first moment may be earlier or later than the moment when the first cell group flows in.

Referring to Figure 2, in this embodiment, at the third moment, the cell group located in the electroporation chamber 2 of the first electrode assembly 11 is the third cell group, and each cell in the third cell group has been subjected to the third A certain period of electroporation treatment of an electrode assembly 11; along the transmission direction, the closer the cells are to the electrode inlet 21 of the first electrode assembly 11, the shorter the electroporation treatment time is, and the closer the cells are to the electrode outlet 22, the shorter the electroporation treatment time is. The longer the treatment time, but the duration of electroporation treatment for each cell in the third cell group does not reach the preset time T. At the third moment, along the transmission direction, the cell group located between the end of the electroporation chamber 2 of the first electrode assembly 11 and the starting end of the electroporation chamber 2 of the second electrode assembly 12 is the first transition group. A transitional group is part of the first cell group. At the third moment, the first transition group has all been electroporated in the electroporation chamber 2 of the first electrode assembly 11 .

Referring to Figure 3, at the second moment, the third cell group all enters the electroporation chamber 2 of the second electrode assembly 12. At this time, each cell that has not been subjected to sufficient electroporation treatment by the first electrode assembly 11 continues to enter the electroporation chamber 2 of the second electrode assembly 11. The electrode assembly 12 is subjected to an electroporation process in the electroporation chamber 2 . Ideally, each cell in the third cell group can be subjected to an electroporation treatment of duration T. At the second moment, all the first transition groups flow through the end of the electroporation chamber 2 of the second electrode assembly 12 and will not be repeatedly electroporated by the second electrode assembly 12 .

In this embodiment, the delay interval between the first moment and the third moment mainly depends on the service life of the first electrode assembly 11 or the length of time it can maintain stable operation. The delay interval between the third moment and the second moment is determined by the time required for the first transition group to all flow through the end of the electroporation chamber 2 of the second electrode assembly 12 .

Referring to FIG. 4 , in this embodiment, at the fourth moment, the cell group located in the electroporation chamber 2 of the second electrode assembly 12 is the fifth cell group. Similarly, each cell in the fifth cell group has been electroporated in the electroporation chamber 2 of the second electrode assembly 12 for a certain period of time, but has not reached the preset time T. At the fourth moment, along the transmission direction, the cell group located between the end of the electroporation chamber 2 of the second electrode assembly 12 and the starting end of the electroporation chamber 2 of the third electrode assembly 13 is the second transition group. The second transition group is part of the second cell group, and at this time, all of the second transition group has been electroporated in the electroporation chamber 2 of the second electrode assembly 12 .

Referring to Figure 5, at the fifth moment, all the fifth cell groups enter the electroporation chamber 2 of the third electrode assembly 13 and continue to be electroporated. Ideally, each cell in the fifth cell group can just Subject to electroporation treatment of duration T. At the fifth moment, all the second transition groups flow through the end of the electroporation chamber 2 of the third electrode assembly 13 and will not be repeatedly processed by the third electrode assembly 13 .

Referring to Figure 6, at the sixth moment, all the fourth cell groups are electroporated by the third electrode assembly 13 and flow out of its electroporation chamber 2. At this time, all the electrode assemblies 1 are closed and continue to wait for all cells. The suspension flows out of the connecting pipe 3.

Similarly, in this embodiment, the delay interval between the second moment and the fourth moment mainly depends on the service life of the second electrode assembly 12 or the length of time it can maintain stable operation. The delay interval between the fifth moment and the sixth moment The delay interval mainly depends on the service life of the third electrode assembly 13 or the length of time it can maintain stable operation. When the specifications of the three groups of electrode assemblies 1 are the same, the above-mentioned working time may be equal. The delay interval between the fourth moment and the fifth moment is determined by the time required for the second transition group to all flow through the end of the electroporation chamber 2 of the third electrode assembly 13 .

It should be noted that the ideal state of the control method in this embodiment is that each cell flowing at a constant speed in the cell suspension can be subjected to electroporation treatment for a duration of T. However, in the actual operation process, due to unavoidable factors such as the non-uniformity of fluid movement, the generation of reaction bubbles, software control errors, etc., it is difficult to ensure that the processing effect of each cell is completely consistent. Therefore, this embodiment aims to provide a better, easy-to-operate serial flow electroporation device and its control method, aiming to improve the consistency of the processing effect of the entire flow electroporation treatment device.

Example 2

Referring to FIGS. 8 to 14 , this embodiment provides a flow electroporation device and a control method thereof. In this embodiment, the structure of the flow electroporation device is basically the same as that in Embodiment 1, and the control method is different. However, the control method is also helpful to realize the flow electroporation device to the large number of people. Continuous and efficient electroporation processing of volumetric cell suspensions.

In this embodiment, there are three groups of electrode assemblies 1. The cell suspension flows sequentially along the transmission direction through the third electrode assembly 13, the second electrode assembly 12 and the electroporation chamber 2 of the first electrode assembly 11 which are connected in series.

Referring to FIG. 8 , in this embodiment, the liquid inlet pipe 31 is connected to the electrode inlet 21 of the third electrode assembly 13 , and the first connecting pipe 33 is connected to the electrode outlet 22 of the third electrode assembly 13 and the second electrode assembly 12 . Between the electrode inlets 21 , the second connecting pipe 34 is connected between the electrode outlet 22 of the second electrode assembly 12 and the electrode inlet 21 of the first electrode assembly 11 , and the liquid outlet pipe 32 is connected with the electrode outlet 22 of the first electrode assembly 11 . In this way, the cell suspension to be treated is continuously input from the sampling container through the liquid inlet pipe 31, and flows through the third electrode assembly 13, the second electrode assembly 12 and the electroporation chamber 2 of the first electrode assembly 11 in sequence along the transmission direction. It flows out from the liquid outlet pipe 32 and is finally collected by the collection container.

It should be noted that the different names of the first electrode assembly 11 , the second electrode assembly 12 , and the third electrode assembly 13 in this article are only for the purpose of distinguishing and explaining the control method, and do not mean that the structure and function of each group of electrode assemblies 1 are different. any limitations.

Similar to Embodiment 1, in the control method of this embodiment, all the cell groups in the cell suspension are also named as the first cell group, the third cell group, the second cell group, and the fifth cell group along the transmission direction. and the fourth cell group. The first cell group is located at the most downstream and enters the liquid inlet pipe 31 first. The fourth cell group is located at the most upstream and enters the liquid inlet pipe 31 last.

Referring to Figures 8 to 14, the control method of the flow electroporation device in this embodiment is described in detail below. The control method includes the following steps in sequence:

S1. Turn on the first electrode assembly 11 at the first moment, see Figure 8; S2. Turn on the second electrode assembly 12 at the second moment, which is later than the first moment, see Figure 9; S3. Close at the third moment. The first electrode assembly 11, the electric field in the electroporation chamber 2 of the first electrode assembly 11 disappears, the third moment is later than the second moment, see Figure 10; S4, the third electrode assembly 13 is turned on at the fifth moment, and at the fifth moment An electric field is formed in the electroporation chamber 2 of the three-electrode assembly 13, and the fifth moment is later than the third moment, see Figure 11; S5. Close the second electrode assembly 12 at the fourth moment, and the electroporation chamber of the second electrode assembly 12 The electric field in 2 disappears, the fourth moment is later than the fifth moment, see Figure 12; S6, the third electrode assembly 13 is closed at the sixth moment, the electric field in the electroporation chamber 2 of the third electrode assembly 13 disappears, the sixth moment The time is later than the fourth time, see Figure 13.

That is to say, in this embodiment, along the transmission direction, the first electrode assembly 11 through which the cell suspension flows last is opened first, then the second electrode assembly 12 in the middle is opened, and the third electrode through which the cell suspension flows first is opened. When the assembly 13 is finally turned on and the previous group of electrode assemblies 1 has not yet been turned off, the latter group of electrode assemblies 1 is turned on in advance.

Referring to Figure 8, in this embodiment, the first moment is the moment when the first cell group has just entered or is about to enter the starting end of the electroporation chamber 2 of the first electrode assembly 11. The liquid is fed from the liquid inlet pipe 31 to the first moment. There is a certain delay between them. At this time, part of the first cell group has flowed through the electroporation chamber 2 of the third electrode assembly 13 or the second electrode assembly 12, but since the first two are not opened, the first cell group will not be subjected to electroporation treatment. .

Referring to Figure 9, in this embodiment, at the second moment, the cell group located in the electroporation chamber 2 of the second electrode assembly 12 is the third cell group, because each cell in the third cell group has been in the A certain distance has flowed in the electroporation chamber 2, so in the next period of time, each cell of the third cell group will be electroporated by the second electrode assembly 12, but the treatment time has not reached the preset time. Time T. At the second moment, along the transmission direction, the cell group located between the end of the electroporation chamber 2 of the second electrode assembly 12 and the starting end of the electroporation chamber 2 of the first electrode assembly 11 is the first transition group. A transitional group is part of the first cell group. At the second time, the first transition group has not yet been subjected to any electroporation treatment.

Referring to Figure 10, at the third moment, the third cell group all enters the electroporation chamber 2 of the first electrode assembly 11. At this time, each cell that has not been subjected to sufficient electroporation treatment by the second electrode assembly 12 continues to enter the electroporation chamber 2 of the first electrode assembly 11. The electrode assembly 11 is subjected to electroporation treatment in the electroporation chamber 2 . Ideally, each cell in the third cell group can be subjected to an electroporation treatment of duration T. At the third moment, all the first transition groups have flowed through the end of the electroporation chamber 2 of the first electrode assembly 11 and been subjected to the electroporation process of the first electrode assembly 11 .

In this embodiment, the delay interval between the first moment and the third moment also depends on the service life of the first electrode assembly 11 or the length of time it can maintain stable operation. The delay interval between the second moment and the third moment is determined by the time required for all the first transition groups to flow through the end of the electroporation chamber 2 of the first electrode assembly 11 .

Referring to FIG. 11 , in this embodiment, at the fifth moment, the cell group located in the electroporation chamber 2 of the third electrode assembly 13 is the fifth cell group. Similarly, each cell in the fifth cell group is subjected to electroporation treatment in the electroporation chamber 2 of the third electrode assembly 13 for a certain period of time, but the preset time T is not reached. At the fifth moment, along the transmission direction, the cell group located between the end of the electroporation chamber 2 of the third electrode assembly 13 and the starting end of the electroporation chamber 2 of the second electrode assembly 12 is the second transition group. The second transition group is part of the second cell group, which has not yet been subjected to any electroporation treatment.

Referring to Figure 12, at the fourth moment, the fifth cell group all enters the electroporation chamber 2 of the second electrode assembly 12 and continues to receive electroporation. Well treatment, ideally, each cell in the fifth cell group can be subjected to an electroporation treatment of duration T. At the fourth moment, all the second transition groups flow through the end of the electroporation chamber 2 of the second electrode assembly 12 and are subjected to the electroporation process of the second electrode assembly 12 .

Referring to Figure 13, at the sixth moment, all the fourth cell groups are electroporated by the third electrode assembly 13 and flow out of its electroporation chamber 2. At this time, all the electrode assemblies 1 are closed and continue to wait for all cells. The suspension flows out of the connecting pipe 3.

Similarly, in this embodiment, the delay interval between the second moment and the fourth moment mainly depends on the service life of the second electrode assembly 12 or the length of time it can maintain stable operation. The delay interval between the fifth moment and the sixth moment The delay interval mainly depends on the service life of the third electrode assembly 13 or the length of time it can maintain stable operation. When the specifications of the three groups of electrode assemblies 1 are the same, the above-mentioned working time may be equal. The delay interval between the fifth moment and the fourth moment is determined by the time required for the second transition group to all flow through the end of the electroporation chamber 2 of the second electrode assembly 12 .

Example 3

Referring to Figure 15, this embodiment provides a flow electroporation device and a control method thereof. In this embodiment, the structure of the flow electroporation device is basically the same as that in Embodiments 1 and 2. The main difference is that in this embodiment there are only two sets of electrode assemblies 1 connected in series through the first connecting tube 33 . After the cell suspension flows in from the liquid inlet pipe 31 , it flows through the electroporation chamber 2 of each group of electrode assemblies 1 in sequence, and finally flows out from the liquid outlet pipe 32 .

For the flow electroporation device of this embodiment, the control method of the flow electroporation device in Embodiment 1 or 2 can be adaptively selected. For example, when the electrode assembly 1 located upstream is the first electrode assembly 11 (left side of the figure) and the electrode assembly 1 located downstream is the second electrode assembly 12 (right side of the figure), the control in Embodiment 1 can be used in sequence. Steps S1 to S4 of the method realize continuous and efficient electroporation of cell suspension.

For another example, when the electrode assembly 1 located downstream is the first electrode assembly 11 and the electrode assembly 1 located upstream is the second electrode assembly 12, steps S1, S2, S3 and S5 of the control method in Embodiment 2 can be used in sequence, Achieve continuous and efficient electroporation of cell suspensions.

In other embodiments, especially when the volume of cell suspension to be processed is larger, the flow electroporation device can also be provided with four or more sets of electrode assemblies 1 connected in series. In this case, those skilled in the art only need Based on the technical concept of the present invention, you can choose to apply the control method in Embodiment 1 or Embodiment 2, and increase the number of repetitions of the corresponding steps accordingly, which will not be described again here.

Example 4

Referring to Figure 16, this embodiment provides a flow electroporation device and a control method thereof. In this embodiment, the flow electroporation device further adds a parallel structure based on the aforementioned series structure.

In this embodiment, the connecting pipeline 3 used to connect multiple electrode assemblies 1 includes a plurality of branches connected in parallel, and each branch is connected to at least one group of electrode assemblies 1 . Specifically, the connecting pipe 3 includes a first pipe group 35, a second pipe group 36 and a first connecting pipe 33. The first pipe group 35 includes a first main road 351 and a plurality of first branch roads 352 that are connected to each other. The second pipe group 36 includes a second main road 361 and a plurality of second branch roads 362 that are connected to each other. In this embodiment, three first branch roads 352 and three second branch roads 362 are specifically shown. Among them, the first main path 351 of the first tube group 35 is connected to the sampling container and is used to input the cell suspension to be processed, and each corresponding first branch path 352 is connected to the electrode inlet of a set of electrode assemblies 1. 21 is connected; the second main path 361 of the second tube group 36 is connected with the collection container and is used to output the processed cell suspension, and each corresponding second branch path 362 is connected to the electrode outlet of a group of electrode assemblies 1 22 connected. The three first branches 352 correspond to the three second branches 362 one-to-one.

In this embodiment, only one set of electrode assemblies 1 is disposed between the two first branches 352 and their corresponding second branches 362; the first electrode assembly 11 and the second electrode assembly 12 are connected by a first The connecting pipes 33 are connected in series and are arranged between the third first branch 352 and its corresponding second branch 362.

For this parallel system, the control method in any one of Embodiments 1 to 3 can be applied to branches with a series structure, so that each branch can achieve continuous and efficient electroporation of cell suspension.

In other embodiments, the number of parallel branches can also be four or more, and different numbers of series or parallel electrode assemblies 1 can also be further provided on each branch. Each branch has a series structure (two and The above branches of the electrode assemblies 1 (connected in series with each other) can all use one of the control methods in any one of the above-mentioned embodiments 1 to 3 to achieve continuous and efficient electroporation processing of the entire flow electroporation device.

The above embodiments are only for illustrating the technical concepts and characteristics of the present invention. Their purpose is to enable those familiar with this technology to understand the content of the present invention and implement it. They are not intended to limit the scope of protection of the present invention. Substantial equivalent changes or modifications shall be included in the protection scope of the present invention.

Claims

A flow electroporation device control method for electroporating a flowing cell suspension, the cell suspension containing a plurality of cell groups arranged sequentially along its own transport direction, each of the cell groups containing A plurality of suspended cells, the plurality of cell groups including at least a first cell group and a second cell group, characterized in that multiple groups of electrode assemblies are sequentially provided along the transmission direction, and each group of the electrode assemblies has an adjustable An electroporation chamber for the circulation of the cell suspension, the plurality of electrode assemblies at least include a first electrode assembly and a second electrode assembly, the first electrode assembly and the second electrode assembly are connected in series, the control Methods include:

The first electrode assembly is turned on, an electric field is formed in the electroporation chamber of the first electrode assembly, and the first cell group is subjected to electroporation treatment in the electroporation chamber of the first electrode assembly. ;

The second electrode assembly is turned on, an electric field is formed in the electroporation chamber of the second electrode assembly, and the second cell group is subjected to electroporation treatment in the electroporation chamber of the second electrode assembly. ;

The first cell group is not subjected to electroporation treatment when flowing through the electroporation chamber of the second electrode assembly, and the second cell group is not subjected to electroporation treatment when flowing through the electroporation chamber of the first electrode assembly. were not subjected to electroporation during treatment.
The flow electroporation device control method according to claim 1, characterized in that when the first cell group flows through the electroporation chamber of the second electrode assembly, the second electrode assembly There is no electric field in the electroporation chamber.
The flow electroporation device control method according to claim 1 or 2, characterized in that when the second cell group flows through the electroporation chamber of the first electrode assembly, the first electrode There is no electric field in the electroporation chamber of the assembly.
The flow electroporation device control method according to any one of claims 1 to 3, characterized in that: turning on the first electrode assembly at a first moment, and turning on the second electrode assembly at a second moment; The above control methods also include:

The first electrode assembly is turned off at a third time, and the electric field in the electroporation chamber of the first electrode assembly disappears. The third time is later than the first time; and the first electrode assembly is turned off at a fourth time. second electrode assembly, the electric field in the electroporation chamber of the second electrode assembly disappears, the fourth time is later than the second time; the first time is not equal to the second time, And/or, the third time is not equal to the fourth time.
The method for controlling a flow electroporation device according to claim 4, wherein the first time is earlier than the second time, and the third time is earlier than the fourth time.
The flow electroporation device control method according to claim 4, wherein the time interval between the first time and the third time is equal to the time interval between the second time and the fourth time. time interval.
The flow electroporation device control method according to claim 4, characterized in that: the cell suspension is transmitted from the first electrode assembly to the second electrode assembly, and the second time is later than the first electrode assembly. Three moments.
The flow electroporation device control method according to claim 4, wherein the cell suspension is transmitted from the second electrode assembly to the first electrode assembly, and the second time is earlier than the first electrode assembly. Three moments.
The flow electroporation device control method according to any one of claims 1 to 8, characterized in that: the plurality of cell groups further include a third cell group, and along the transmission direction, the third cell group Located between the first cell group and the second cell group, the third cell group is electroporated while flowing through the electroporation chamber of the first electrode assembly. Groups are also electroporated while flowing through the electroporation chamber of the second electrode assembly.
The flow electroporation device control method according to any one of claims 1 to 9, characterized in that: the time for each cell in the first cell group to be electroporated is the same as the time for each cell in the second cell group to be subjected to electroporation treatment. Each cell was electroporated for an equal amount of time.
The flow electroporation device control method according to any one of claims 1-3 and 9-10, characterized in that: the plurality of groups of electrode assemblies further include a third electrode assembly, and the cell suspension is along the The transmission direction flows through the electroporation chamber of the first electrode assembly, the second electrode assembly and the third electrode assembly in sequence; the control method includes the following steps in sequence: S1. Turn on all the electroporation chambers at the first moment. The first electrode assembly; S2. Turn off the first electrode assembly at a third moment, and the electric field in the electroporation chamber of the first electrode assembly disappears, and the third moment is later than the first moment. ; S3. Turn on the second electrode assembly at the second moment, and the second moment is later than the third moment; S4. Close the second electrode assembly at the fourth moment, and all the parts of the second electrode assembly The electric field in the electroporation chamber disappears, and the fourth moment is later than the second moment; S5. Turn on the third electrode assembly at the fifth moment, and the electroporation chamber of the third electrode assembly An electric field is formed in the electroporation chamber, and the fifth moment is later than the fourth moment; S6. Turn off the third electrode assembly at the sixth moment, and the electric field in the electroporation chamber of the third electrode assembly disappears, so The sixth moment is later than the fifth moment.
The flow electroporation device control method according to claim 11, characterized in that:

At the third moment, along the transmission direction, the end of the electroporation chamber located at the first electrode assembly and the second electrode The cell group between the starting ends of the electroporation chamber of the assembly is a first transition group; at the second moment, along the transport direction, all of the first transition group flows through the second The end of the electroporation chamber of the electrode assembly; at the fourth moment, along the transmission direction, the end of the electroporation chamber of the second electrode assembly is located between the end of the electroporation chamber of the third electrode assembly and The cell group between the starting ends of the electroporation chamber is a second transition group; at the fifth moment, along the transmission direction, all of the second transition group flows through the third electrode assembly end of the electroporation chamber.
The flow electroporation device control method according to any one of claims 1-3 and 9-10, characterized in that: the plurality of groups of electrode assemblies further include a third electrode assembly, and the cell suspension is along the The transmission direction flows through the electroporation chamber of the third electrode assembly, the second electrode assembly and the first electrode assembly in sequence; the control method includes the following steps in sequence: S1. Turn on all the electroporation chambers at the first moment. The first electrode assembly; S2, turn on the second electrode assembly at a second time, the second time being later than the first time; S3, turn off the first electrode assembly at a third time, the third time The electric field in the electroporation chamber of an electrode assembly disappears, and the third time is later than the second time; S4. Turn on the third electrode assembly at the fifth time, and all the components of the third electrode assembly An electric field is formed in the electroporation chamber, and the fifth time is later than the third time; S5. Close the second electrode assembly at the fourth time, and the second electrode assembly is in the electroporation chamber. The electric field disappears, and the fourth moment is later than the fifth moment; S6. Turn off the third electrode assembly at the sixth moment, and the electric field in the electroporation chamber of the third electrode assembly disappears, so The sixth moment is later than the fourth moment.
The flow electroporation device control method according to claim 13, characterized in that:

At the second moment, along the transport direction, all the elements located between the end of the electroporation chamber of the second electrode assembly and the starting end of the electroporation chamber of the first electrode assembly The cell group is a first transition group; at the third moment, along the transmission direction, all the first transition group flows through the end of the electroporation chamber of the first electrode assembly; at the At the fifth moment, along the transmission direction, the cell group is located between the end of the electroporation chamber of the third electrode assembly and the starting end of the electroporation chamber of the second electrode assembly. is the second transition group; at the fourth moment, along the transport direction, all of the second transition group flows through the end of the electroporation chamber of the second electrode assembly.
A flow electroporation device used for electroporation of cell suspension, characterized in that: the flow electroporation device includes multiple groups of electrode assemblies, and each group of the electrode assemblies has an electrode assembly for the cell suspension. An electroporation chamber with liquid flow, the plurality of electrode assemblies at least include a first electrode assembly and a second electrode assembly, the first electrode assembly and the second electrode assembly are connected in series, the flow electroporation device further It includes a control system for controlling the first electrode assembly and the second electrode assembly to be turned on at different times.
A flow electroporation device used for electroporation of cell suspension, characterized in that: the flow electroporation device includes multiple groups of electrode assemblies, and each group of the electrode assemblies has an electrode assembly for the cell suspension. An electroporation chamber with liquid flow, the plurality of electrode assemblies at least include a first electrode assembly and a second electrode assembly, the first electrode assembly and the second electrode assembly are connected in series, the flow electroporation device further A control system is included, and the control system is used to control the first electrode assembly and the second electrode assembly to close at different times.
The flow electroporation device according to claim 16, wherein the control system is further used to control the first electrode assembly and the second electrode assembly to be turned on at different times.
The flow electroporation device according to any one of claims 15 to 17, characterized in that: the plurality of groups of electrode assemblies further include a third electrode assembly, the first electrode assembly, the second electrode assembly and The third electrode assembly is connected in series, the second electrode assembly is located between the first electrode assembly and the third electrode assembly, and the control system is also used to control the opening and closing of the third electrode assembly. .
A flow electroporation device used for electroporation of cell suspension, characterized in that: the flow electroporation device includes multiple groups of electrode assemblies and connecting pipelines, and each group of electrode assemblies has a The electroporation chamber through which the cell suspension circulates; a plurality of the electrode assemblies are connected through the connecting pipeline, the connecting pipeline includes a plurality of branches connected in parallel, each of the branches is connected to at least one Set the electrode assembly connections.
The flow electroporation device according to claim 19, wherein the plurality of groups of electrode assemblies at least include a first electrode assembly and a second electrode assembly connected in series to one of the branches.
The flow electroporation device according to any one of claims 15 to 20, characterized in that: the control system uses the control method according to any one of claims 1 to 14 to control the flow electroporation. device.