CN117660176A - Flow electroporation component - Google Patents
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- CN117660176A CN117660176A CN202211026908.0A CN202211026908A CN117660176A CN 117660176 A CN117660176 A CN 117660176A CN 202211026908 A CN202211026908 A CN 202211026908A CN 117660176 A CN117660176 A CN 117660176A
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- 238000004520 electroporation Methods 0.000 title claims abstract description 153
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Abstract
The invention discloses a flow electroporation component, which comprises a main liquid inlet, a main liquid outlet, a connecting pipeline and at least two groups of electrode components. Each group of electrode assemblies comprises two plane electrodes arranged at intervals, an electroporation chamber extending along the up-down direction is arranged between the two plane electrodes, the electroporation chamber is provided with an electrode inlet and an electrode outlet, the electrode inlet is positioned at the bottom end of the electroporation chamber, the electrode outlet is positioned at the top end of the electroporation chamber, and when the cell suspension flows through the electroporation chamber, the cell suspension flows from the electrode inlet to the corresponding electrode outlet. Among the multiple groups of electrode assemblies, the electrode inlet of one group of electrode assemblies is communicated with the main liquid inlet, the electrode outlet of the other group of electrode assemblies is communicated with the main liquid outlet, and the electroporation chambers of different electrode assemblies are communicated through connecting pipelines. The flow electroporation component of the invention remarkably improves electroporation processing capacity, and can continuously, efficiently and stably carry out electroporation processing on more cell suspensions.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a flow electroporation component.
Background
Electroporation (electric corporation) is a technique that uses an electric field to act on a cell membrane to make the cell membrane generate micropores through which foreign molecules such as DNA, RNA, proteins can pass, and these micropores are instantaneously generated by the electric field and can be recovered as the electric field disappears, thereby avoiding permanent damage to cells. Various exogenous molecules such as DNA, RNA, proteins, carbohydrates, dyes, viral particles, etc. can be introduced into prokaryotic or eukaryotic cells using electroporation techniques, known as Electrotransfection; the electrotransfection technology is widely applied to the technical fields of various high-end biological medicines such as antibody protein production, in Vitro Diagnosis (IVD) reagent raw material production, immune cell gene editing and gene modification, and the like, and is an important technology for delivering substances in cells.
There are various methods for electroporation of cells, among which, the flow electroporation technique can continuously electroporate a flowing cell-containing suspension (hereinafter referred to as a cell suspension), which is highly efficient and particularly suitable for batch processing of large-volume cell suspensions. A common flow electroporation component mainly comprises a pair of planar electrodes for generating an electric field, and a chamber between the two planar electrodes, through which a cell suspension can be shocked while flowing, and electroporation is generated, so that exogenous molecules in the cell suspension can enter target cells. Generally, in order to ensure that the cell suspension is not contaminated, the whole electroporation process needs to be performed in a strictly sterile environment, so that the storage container, the transmission pipeline, the planar electrode and the like in direct contact with the cell suspension in each processing batch are disposable consumables and cannot be reused.
In the prior art, the flow electroporation component mostly has the problem of low cell treatment efficiency (such as electrotransfection efficiency), and when the treatment volume of the cell suspension is large and the electrode needs to work for a long time, the problem of reduction of the electrotransfection efficiency and the cell survival rate with the increase of time is particularly obvious. On the one hand, the area of each planar electrode is not too large, the distance between the two electrodes cannot be too large, otherwise, the voltage and the power of a power supply are obviously increased, the economic cost is increased, a large number of cells die due to the problems of too high voltage, serious electrode heating and the like, and the electroporation chamber of a pair of planar electrodes with small area and small distance is smaller, so that the volume of cell suspension which can be processed in unit time is limited. On the other hand, as the working time of the electrode increases, dead cells, cell fragments and other solid matters in the cell suspension can be gradually attached to the surface of the electrode, a layer of coating film is formed on the surface of the electrode, the conductivity of the electrode is weakened, the electric field in the cell suspension is weakened, and the electric transfection efficiency is lower and lower; meanwhile, as the working time of the electrode increases, the surface temperature of the electrode gradually increases, so that a large number of cells die, and the cell survival rate is reduced. In addition, the high voltage causes the cell suspension to generate a stronger water electrolysis phenomenon when receiving electric shock, generates a large number of bubbles, brings the problems of uneven liquid flow, unstable flow speed and the like, breaks the uniformity of an electric field, influences the controllability of the times of receiving the electric shock of cells, and further causes the reduction of the cell electrotransfection efficiency.
In view of the above, the existing flow electroporation components are not capable of performing continuous, efficient and stable electroporation of large volumes of cell suspensions.
Disclosure of Invention
The invention aims at solving the problems existing in the prior art and provides a flow electroporation component which can carry out continuous, efficient and stable electroporation treatment on a large-volume cell suspension.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a flow electroporation component for electroporation treatment of cell suspension comprises a main liquid inlet, a main liquid outlet, a connecting pipeline and at least two groups of electrode components, wherein,
each group of electrode assemblies comprises two plane electrodes which are arranged at intervals, an electroporation chamber extending along the up-down direction is arranged between the two plane electrodes, the electroporation chamber is provided with an electrode inlet and an electrode outlet, the electrode inlet is positioned at the bottom end of the electroporation chamber, the electrode outlet is positioned at the top end of the electroporation chamber, and when the cell suspension flows through the electroporation chamber, the cell suspension flows from the electrode inlet to the corresponding electrode outlet;
among the multiple groups of electrode assemblies, the electrode inlet of one group of electrode assemblies is communicated with the main liquid inlet, the electrode outlet of the other group of electrode assemblies is communicated with the main liquid outlet, and the electroporation chambers of different electrode assemblies are communicated through the connecting pipelines.
In some embodiments, the electrode outlet of one of the electrode assemblies is connected to the electrode inlet of another of the electrode assemblies by the connecting line.
In some embodiments, the connecting line comprises one or more connecting tube segments, along the transport direction of the cell suspension: and one connecting pipe section is connected between every two adjacent electrode assemblies, and the connecting pipe sections are respectively connected with the electrode inlet of the electrode assembly positioned at the front side and the electrode outlet of the electrode assembly positioned at the rear side. Thus, the electrode assemblies are connected in series, the cell suspension can sequentially flow through each electroporation chamber, and the electrode assemblies can work in a time-sharing or simultaneous manner so as to improve the processing capacity of the flow electroporation assembly.
In some embodiments, along the transport direction of the cell suspension: in each adjacent two sets of the electrode assemblies, the electrode inlet of the electrode assembly located at the front side has a lower height than the electrode outlet of the electrode assembly located at the rear side. The height of the flow electroporation component is controlled, the structure is more compact, and the occupied space is smaller.
In some embodiments, along the transport direction of the cell suspension: every two adjacent groups of electrode assemblies are arranged at intervals in a horizontal plane, the horizontal distance between the two groups of electrode assemblies is D, and the radius of curvature at any position of the axial lead of the connecting pipe section between the two groups of electrode assemblies is larger than D/4. The connecting pipe sections have small curvature change, so that the cell suspension is not easy to bend to block or even block.
In order to improve the working reliability of the flow electroporation component, the invention needs to ensure that each bent and extended connecting pipe section cannot be bent in the working process, otherwise, the smoothness of the cell suspension flow in the connecting pipeline is influenced. In some embodiments, each connecting pipe section of the invention adopts a flexible hose, and when the curvature of the connecting pipe section is too large, local bending is easy to occur due to tiny disturbance, so that the flow of cell suspension in the connecting pipe is blocked or even blocked. If the flow of the cell suspension in the pipeline is blocked or even blocked during the electroporation process, part of cells can die due to repeated electric shock for a long time, and the cells can be killed due to toxicity caused by too much substances introduced into the cells.
In some embodiments, the radius of curvature of the connecting tube segment at least at some locations along the axis is approximately equal to D/2. So that the connecting line as a whole has a smaller curvature (i.e. a larger radius of curvature). In some embodiments, each connecting tube section extends approximately in an S shape, and fills the space between the two groups of electrode assemblies, so that the flow of the cell suspension is smoother, and bending of the connecting tube is effectively avoided.
In some embodiments, along the transport direction of the cell suspension: every two adjacent electrode assemblies are arranged at intervals along a horizontal first direction, the connecting pipe section between the two electrode assemblies is provided with a first rotary section, a second rotary section and a third rotary section, the first rotary section, the second rotary section and the third rotary section are sequentially connected from back to front along the transmission direction of the cell suspension, and are positioned on the same side of the second rotary section along the first direction. The arrangement of the first, second and third turnaround sections defines the direction of extension of the connecting tube sections and also contributes to an increase of the radius of curvature of the connecting tube sections.
In some embodiments, along the transport direction of the cell suspension: in each two adjacent groups of electrode assemblies, the electrode assembly positioned at the front side is a front electrode assembly, the electrode assembly positioned at the rear side is a rear electrode assembly, the connecting pipe section between the front electrode assembly and the rear electrode assembly is provided with at least two second revolving sections, the second revolving section close to the front electrode assembly in the two second revolving sections is a front second revolving section, and the second revolving section close to the rear electrode assembly is a rear second revolving section; along the first direction: the front second turnaround section is adjacent to the rear electrode assembly and the rear second turnaround section is adjacent to the front electrode assembly. Thereby further limiting the approximately S-shaped extension of the connecting pipe section and effectively avoiding the folding of the connecting pipeline.
In some embodiments, the connecting pipeline comprises a first pipe group and a second pipe group, the first pipe group comprises a first main pipeline and at least two first branches which are communicated with each other, the first main pipeline is connected with the main liquid inlet, and the electrode inlets of each group of the electrode assemblies are correspondingly connected with one first branch; the second tube group comprises a second main path and at least two second branches which are communicated with each other, the second main path is connected with the main liquid outlet, and each electrode outlet of each electrode assembly is correspondingly connected with one second branch. Thus, the electrode assemblies are connected in parallel, cell suspension can flow through each electroporation chamber at the same time, and the electrode assemblies can work simultaneously or in a time-sharing manner so as to improve the processing capacity of the flow electroporation assembly.
In some embodiments, the first tube bank is located below the second tube bank, the first main is located below all of the first branches, and the second main is located above all of the second branches. Thereby optimizing the overall layout of the connecting lines, and the cell suspension can flow through the flow electroporation component from bottom to top as a whole.
In some embodiments, the primary liquid inlet is located below the primary liquid outlet. So that the cell suspension flows through the flow electroporation component from bottom to top as a whole.
In some embodiments, the axis of the main liquid inlet and the axis of the main liquid outlet extend in the vertical direction respectively, and the axis of the main liquid inlet and the axis of the main liquid outlet extend in a collinear manner. Thereby the main liquid inlet is aligned with the main liquid outlet from top to bottom, the structure is neat, and the installation and the use of the flow electroporation component are convenient.
In some embodiments, the electrode inlets of different ones of the electrode assemblies are located at the same height and the electrode outlets of different ones of the electrode assemblies are located at the same height. The flow electroporation component is thus more compact and less bulky.
In some embodiments, in the up-down direction: the length of the electroporation chambers of all the electrode assemblies is equal, and the cross-sectional areas of the electroporation chambers of all the electrode assemblies at any positions are equal. Thereby further ensuring consistency of electroporation processing effects of the cell suspension in the different electroporation chambers.
In some embodiments, the electrode assemblies are arranged at intervals along a horizontal first direction, and the axes of the electroporation chambers of all the electrode assemblies and the axes of the connecting pipelines are located in the same plane. The flow electroporation component is flat in structure, small in occupied space and beneficial to modular manufacturing.
In some embodiments, the axis of the main liquid inlet, the axis of the main liquid outlet, the axis of the connecting pipeline, and the axes of the electroporation chambers of all the electrode assemblies are all located in the same vertical plane. Thereby further facilitating modular fabrication of the flow electroporation component.
Due to the application of the technical scheme, the flow electroporation component provided by the invention comprises at least two groups of electrode components, electroporation chambers of different electrode components are communicated through connecting pipelines, and the multiple groups of electrode components can be connected in a plurality of modes such as series connection, parallel connection or series-parallel connection mixing and the like by different connecting modes among the connecting pipelines. The cell suspension flowing in from the main liquid inlet can sequentially or simultaneously flow through different electrode assemblies, the volume of the cell suspension required to be treated by each group of electrode assemblies is reduced by times, and when the service life of each pair of plane electrodes is unchanged, the technical scheme of the invention obviously improves the treatment capacity of the flow electroporation assembly. Compared with the prior art, the flow electroporation component can carry out electroporation treatment on more cell suspensions with higher and more stable electrotransfection efficiency in the same time; and/or the flow electroporation component is capable of maintaining a higher, more stable electroporation efficiency for a longer period of time for more cell suspension. Furthermore, each group of electroporation chambers in the invention adopts the flow direction of the lower inlet and the upper outlet, so that the part of the electroporation chamber through which the cell suspension flows can be always filled with the liquid, the cell suspension can continuously and uniformly flow through the electroporation chamber, and bubbles generated in the electroporation process can be rapidly discharged through an electrode outlet at the top end of the electroporation chamber, and cannot accumulate in the electroporation chamber to influence the electroporation effect. Compared with the scheme of adopting the flow direction of lower inlet and upper outlet in a single electroporation chamber, the invention further organically combines a plurality of groups of electrode assemblies containing the electroporation chambers, ensures that each electroporation chamber keeps the flow direction of lower inlet and upper outlet through reasonable layout of connecting pipelines, ensures that cell suspension can be stably and efficiently subjected to electroporation in each different electroporation chamber, further improves the overall electroporation efficiency and cell survival rate of the flow electroporation assembly, and realizes the long-time stable electroporation treatment on the cell suspension with larger volume.
Drawings
In order to more clearly illustrate the technical solution of the present invention, the drawings used in the description of the embodiments will be briefly described.
FIG. 1 is a schematic diagram of a flow electroporation component according to embodiment 1 of the present invention;
FIG. 2 is a schematic diagram of a partial structure of the flow electroporation component of example 1;
FIG. 3 is a schematic diagram of the flow electroporation component of example 2;
FIG. 4 is a schematic diagram of the flow electroporation component of example 3;
FIG. 5 is a schematic diagram of the flow electroporation component of example 4;
FIG. 6 is a schematic diagram of the flow electroporation component of example 5;
wherein: 1. a main liquid inlet; 11. a liquid inlet pipe; 2. a main liquid outlet; 21. a liquid outlet pipe;
3. a connecting pipeline; 31. connecting pipe sections; 311. a first turnaround section; 311a, a front first turnaround section; 311b, a rear first swing section; 312. a second turnaround section; 312a, a front second turnaround section; 312b, a rear second turnaround section; 313. a third swivel segment; 313a, a front third swing section; 313b, a rear third swing section; 32. a first tube group; 321. a first main path; 322. a first branch; 33. a second tube group; 331. a second main path; 332. a second branch;
4. an electrode assembly; 4a, front electrode assembly; 4b, a rear electrode assembly; 41. a planar electrode; 42. an electroporation chamber; 421. an electrode inlet; 422. and an electrode outlet.
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to the attached 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 FIG. 1, the present embodiment provides a flow electroporation component for electroporation of a cell suspension. The flow direction of the cell suspension during normal operation of the flow electroporation component is shown by solid arrows. The flow electroporation component comprises a main liquid inlet 1, a main liquid outlet 2, a connecting pipeline 3 and at least two groups of electrode components 4, wherein in the embodiment, the electrode components 4 are specifically provided with three groups. In the figure, the three-dimensional coordinate system is established by taking the flow electroporation component as a reference, and the three-dimensional coordinate system comprises an up-down direction Z, a first direction X and a second direction which are perpendicular to the paper surface, wherein the second direction is a direction perpendicular to the paper surface, but the inventive structure of the flow electroporation component is mainly embodied in the up-down direction Z and the first direction X (namely, the part shown in the figure), so that the structure of the flow electroporation component in the second direction is not specifically described in the embodiment. When the flow electroporation component is in normal operation, the up-down direction Z extends in a vertical direction and the first direction X extends in a horizontal direction. The definition of orientation herein is for convenience of description and understanding only, and the orientation should not be construed as limiting the invention since the flow electroporation component may be placed in any orientation when in the inactive state.
In this embodiment, each set of electrode assemblies 4 includes two planar electrodes 41 disposed at intervals, an electroporation chamber 42 extending along an up-down direction Z is disposed between the two planar electrodes 41, the electroporation chamber 42 has an electrode inlet 421 and an electrode outlet 422, the electrode inlet 421 is located at a bottom end of the electroporation chamber 42 along the up-down direction Z, and the electrode outlet 422 is located at a top end of the electroporation chamber 42. As the cell suspension flows through the electroporation chamber 42, the cell suspension flows from the electrode inlet 421 to the corresponding electrode outlet 422, i.e., the cell suspension flows through each electroporation chamber 42 in a downward, upward, and downward flow direction. In the present embodiment, two planar electrodes 41 in each group of electrode assemblies 4 are each disposed at intervals along the first direction X. Along the up-down direction Z: the length of the electroporation chambers 42 of all the electrode assemblies 4 is equal, and the cross-sectional areas of the electroporation chambers 42 of all the electrode assemblies 4 at any positions are equal. In this embodiment, the geometry of the three sets of electrode assemblies 4 is identical to ensure uniformity of electroporation treatment of the cell suspension in the different electrode assemblies 4.
Referring to fig. 1, in a plurality of electrode assemblies 4, an electrode inlet 421 of one electrode assembly 4 is communicated with a main liquid inlet 1, an electrode outlet 422 of another electrode assembly 4 is communicated with a main liquid outlet 2, and electroporation chambers 42 of different electrode assemblies 4 are communicated through a connecting pipeline 3. As can be seen from the figure, in the multiple electrode assemblies 4, the electrode outlet 422 of one electrode assembly 4 is connected to the electrode inlet 421 of another electrode assembly 4 through the connecting pipeline 3, in this embodiment, the three electrode assemblies 4 are all connected in series, and the cell suspension flows through each electrode assembly 4 in sequence from left to right. The side close to the main liquid outlet 2 is herein referred to as the "front side" of the transport direction of the cell suspension, and the side close to the main liquid inlet 1 is referred to as the "rear side" of the transport direction of the cell suspension.
Referring to fig. 1, the electrode inlet 421 of the leftmost electrode assembly 4 is in communication with the main liquid inlet 1, specifically through the liquid inlet pipe 11, as viewed in the drawing; the electrode outlet 422 of the rightmost electrode assembly 4 communicates with the main liquid outlet 2, in particular via the liquid outlet pipe 21. In this embodiment, the main liquid inlet 1 is located below the main liquid outlet 2, and the axis of the main liquid inlet 1 and the axis of the main liquid outlet 2 extend along the up-down direction Z, respectively, and the axis of the main liquid inlet 1 and the axis of the main liquid outlet 2 extend in a collinear manner. Specifically, the liquid inlet pipe 11 and the liquid outlet pipe 21 are hoses, the liquid outlet pipe 21 shown in the figure extends vertically upwards, the liquid inlet pipe 11 bends downwards from the electrode inlet 421 of the leftmost electrode assembly 4 and extends a distance along the first direction X and then extends vertically downwards, and the design mainly enables the main liquid inlet 1 and the main liquid outlet 2 to approach each other in the first direction X, so that the practical installation and use of the flow electroporation assembly are facilitated.
In this example, along the transport direction of the cell suspension: the plurality of electrode assemblies 4 are arranged in sequence, every two adjacent electrode assemblies 4 are arranged at intervals in a horizontal plane, and in every two adjacent electrode assemblies 4, the height of an electrode inlet 421 of the electrode assembly 4 positioned at the front side is lower than the height of an electrode outlet 422 of the electrode assembly 4 positioned at the rear side. Further, in this embodiment, along the transport direction of the cell suspension, the multiple groups of electrode assemblies 4 are arranged at intervals along the first direction X, and the electrode inlets 421 of different electrode assemblies 4 are located at the same height, and the electrode outlets 422 of different electrode assemblies 4 are located at the same height. Referring to fig. 1, along the transport direction of the cell suspension, the horizontal distance between each two adjacent electrode assemblies 4 is D, in this embodiment, the horizontal distance between the two electrode assemblies 4 on the left side is equal to the horizontal distance between the two electrode assemblies 4 on the right side, i.e., three electrode assemblies 4 are equidistantly arranged along the first direction X.
In this embodiment, the above arrangement of the plurality of groups of electrode assemblies 4 can reduce the overall height of the flow electroporation assembly, and the flow electroporation assembly has compact structure and regular arrangement, which is beneficial to modular manufacture of the flow electroporation assembly. In other embodiments, the multiple sets of electrode assemblies 4 may not be perfectly aligned in the first direction X, and the horizontal distances between different electrode assemblies 4 may not be perfectly equal.
Referring to fig. 1, in this embodiment, the connecting line 3 comprises one or more connecting tube sections 31. A connecting tube section 31 is connected between each adjacent two sets of electrode assemblies 4 along the transport direction of the cell suspension. The front end of each connection pipe section 31 is connected to the electrode inlet 421 of the electrode assembly 4 located on the front side, and the rear end of each connection pipe section 31 is connected to the electrode outlet 422 of the electrode assembly 4 located on the rear side. Specifically, in this embodiment, two connecting tube sections 31 are provided in total between the three groups of electrode assemblies 4, and the lengths and shapes of the two connecting tube sections 31 are the same. In this embodiment, the axes of the electroporation chambers 42 of all the electrode assemblies 4 and the axes of the connecting pipelines 3 are located in the same plane, and further the axes of the main liquid inlet 1, the axes of the main liquid outlet 2, the axes of the connecting pipelines 3 and the axes of the electroporation chambers 42 of all the electrode assemblies 4 are located in the same vertical plane, so that the flow electroporation assembly has flat structure and small occupied space, is beneficial to modular manufacturing and is convenient to install.
Referring to fig. 1, in this embodiment, the direction of transport of the cell suspension is: the radius of curvature r at any position of the axis of the connecting tube segment 31 between each adjacent two sets of electrode assemblies 4 is greater than D/4, and further the radius of curvature r at least a part of the axis of the connecting tube segment 31 is substantially equal to D/2. More specifically, the axes of the upper and lower halves of each connecting tube segment 31 are each located approximately on an arc having a radius of curvature r equal to about D/2, with smooth junctions between the two arcs.
Referring to fig. 1, in this embodiment, the direction of transport of the cell suspension is: the connecting tube sections 31 between every two adjacent electrode assemblies 4 are respectively provided with a first rotary section 311, a second rotary section 312 and a third rotary section 313, and the first rotary section 311, the second rotary section 312 and the third rotary section 313 are sequentially connected from back to front along the transmission direction of the cell suspension. The first turn section 311 is located on the same side of the second turn section 312 as the third turn section 313 along the first direction X. Further, see fig. 2, along the transport direction of the cell suspension: in each two adjacent electrode assemblies 4, the electrode assembly 4 positioned at the front side is a front electrode assembly 4a, and the electrode assembly 4 positioned at the rear side is a rear electrode assembly 4b; the connecting tube section 31 between the front electrode assembly 4a and the rear electrode assembly 4b has at least two second turning sections 312, of which the second turning section 312 near the front electrode assembly 4a is a front second turning section 312a and the second turning section 312 near the rear electrode assembly 4b is a rear second turning section 312b. In the first direction X, the front second turnaround section 312a is adjacent to the rear electrode assembly 4b, and the rear second turnaround section 312b is adjacent to the front electrode assembly 4a. Further shown in fig. 2 are a front first turn section 311a located on the rear side of the front second turn section 312a, a front third turn section 313a located on the front side of the front second turn section 312a, and a rear first turn section 311b located on the rear side of the rear second turn section 312b, a rear third turn section 313b located on the front side of the rear second turn section 312b. As can be seen, the connecting tube segment 31 is actually "S" shaped (or mirrored "S" shaped) in curvature and extends, the front first turnaround section 311a and the rear third turnaround section 313b being considered to be the same part. In combination with the foregoing description about the radius of curvature r of the axis of the connecting tube segment 31, in the limited space between two adjacent electrode assemblies 4, the curvature is minimized at any position of the axis of the connecting tube segment 31 in this embodiment, so that the possibility of local bending of the connecting tube segment 31 due to the overlarge curvature of the axis of the connecting tube segment is avoided to the greatest extent, and the smoothness of the cell suspension flow is ensured.
Example 2
Referring to fig. 3, this embodiment provides a flow electroporation component which is substantially the same as embodiment 1, mainly differing in that the flow electroporation component in this embodiment includes only two sets of electrode assemblies 4, and accordingly, the connection pipe 3 includes only one connection pipe section 31, and a front end portion of the connection pipe section 31 is connected to an electrode inlet 421 of the electrode assembly 4 located on the front side, and a rear end portion thereof is connected to an electrode outlet 422 of the electrode assembly 4 located on the rear side.
In other embodiments, the tandem flow electroporation module may also include four or more electrode assemblies 4, and the number of connecting tube sections 31 should be increased, and when the electrode assemblies 4 have n groups (n is an integer greater than or equal to 2), the number of connecting tube sections 31 is n-1.
Example 3
Referring to fig. 4, this embodiment provides a flow electroporation component, which is substantially the same as embodiment 2, and is mainly different in that the axis of the main liquid inlet 1 and the axis of the main liquid outlet 2 of the flow electroporation component in this embodiment do not extend in a collinear manner.
In this embodiment, the liquid inlet pipe 11 and the liquid outlet pipe 21 extend along the vertical direction Z, respectively, and the liquid inlet pipe 11 is not bent, so that the axis of the main liquid inlet 1 and the axis of the main liquid outlet 2 are parallel to each other and are arranged at intervals along the first direction X.
In other embodiments, the inlet tube 11 and the outlet tube 21 may be arranged in other extending directions which are easy to think, depending on the actual installation and use of the flow electroporation component, for example, when the flow electroporation component is required to be connected with an external housing or a pipeline in a matching manner.
Example 4
Referring to fig. 5, this embodiment provides a flow electroporation component, in which the arrangement of the electrode assembly 4 is substantially the same as that of embodiment 1, and the main difference is that the specific arrangement of the connecting line 3 and the flow pattern of the cell suspension are different.
In this embodiment, the flow electroporation component also includes three groups of electrode assemblies 4 arranged at intervals along the first direction X, and the heights of the three groups of electrode assemblies 4 are equal. Referring to fig. 5, in this embodiment, the separate liquid inlet pipe 11 and liquid outlet pipe 21 are not provided. The connecting line 3 comprises a first tube set 32 and a second tube set 33. The first tube set 32 includes a first main path 321 and at least two first branches 322, where the first main path 321 is connected to the main liquid inlet 1, and the electrode inlets 421 of each set of electrode assemblies 4 are correspondingly connected to one first branch 322. Similarly, the second tube group 33 includes a second main path 331 and at least two second branches 332 that are in communication with each other, the second main path 331 is connected to the main liquid outlet 2, and the electrode outlets 422 of each group of electrode assemblies 4 are respectively connected to one of the second branches 332. In this embodiment, corresponding to the three groups of electrode assemblies 4, there are three first branches 322 and three second branches 332. In the present embodiment, the first tube set 32 is located below the second tube set 33 as a whole, and the first main path 321 is located below all the first branches 322, and the second main path 331 is located above all the second branches 332.
Thus, the three groups of electrode assemblies 4 in the embodiment are connected in parallel, and the cell suspension flowing in from the main liquid inlet 1 can be divided into three paths in the first tube group 32, and respectively flow to the three groups of electrode assemblies 4 from bottom to top, and electroporation treatment is performed at the same time; the cell suspensions of the three groups of electrode assemblies 4 after electroporation are converged from bottom to top through the second tube group 33, and finally flow out of the main liquid outlet 2. The parallel arrangement can improve the working efficiency of the flow electroporation component and the cell electrotransfection treatment efficiency.
Example 5
Referring to fig. 6, this embodiment provides a flow electroporation component which is substantially the same as embodiment 5, and is mainly different in that the flow electroporation component in this embodiment includes only two sets of electrode assemblies 4, correspondingly, the first tube set 32 includes a first main path 321 and two first branches 322 which are communicated with each other, and the second tube set 33 includes a second main path 331 and two second branches 332 which are communicated with each other.
In other embodiments, the parallel flow electroporation component may also include four or more electrode assemblies 4, and the number of the first branches 322 and the second branches 332 is increased and kept consistent with the number of the electrode assemblies 4.
In summary, the flow electroporation component provided by the invention can significantly improve electroporation processing capability, and is particularly suitable for continuous, efficient and stable electroporation processing of a large-volume cell suspension.
The above embodiments are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the content of the present invention and to implement the same, but are not intended to limit the scope of the present invention, and all equivalent changes or modifications made according to the spirit of the present invention should be included in the scope of the present invention.
Claims (15)
1. A flow electroporation component for electroporation of a cell suspension, comprising: the flow electroporation component comprises a main liquid inlet, a main liquid outlet, a connecting pipeline and at least two groups of electrode components, wherein,
each group of electrode assemblies comprises two plane electrodes which are arranged at intervals, an electroporation chamber extending along the up-down direction is arranged between the two plane electrodes, the electroporation chamber is provided with an electrode inlet and an electrode outlet, the electrode inlet is positioned at the bottom end of the electroporation chamber, the electrode outlet is positioned at the top end of the electroporation chamber, and when the cell suspension flows through the electroporation chamber, the cell suspension flows from the electrode inlet to the corresponding electrode outlet;
among the multiple groups of electrode assemblies, the electrode inlet of one group of electrode assemblies is communicated with the main liquid inlet, the electrode outlet of the other group of electrode assemblies is communicated with the main liquid outlet, and the electroporation chambers of different electrode assemblies are communicated through the connecting pipelines.
2. The flow electroporation assembly of claim 1, wherein: and in the electrode assemblies, the electrode outlet of one group of the electrode assemblies is connected with the electrode inlet of the other group of the electrode assemblies through the connecting pipeline.
3. The flow electroporation assembly of claim 1, wherein: the connecting line comprises one or more connecting tube sections along the transport direction of the cell suspension: and one connecting pipe section is connected between every two adjacent electrode assemblies, and the connecting pipe sections are respectively connected with the electrode inlet of the electrode assembly positioned at the front side and the electrode outlet of the electrode assembly positioned at the rear side.
4. A flow electroporation component as claimed in claim 3, wherein: along the transport direction of the cell suspension: in each adjacent two sets of the electrode assemblies, the electrode inlet of the electrode assembly located at the front side has a lower height than the electrode outlet of the electrode assembly located at the rear side.
5. The flow electroporation assembly of claim 4, wherein: along the transport direction of the cell suspension: every two adjacent groups of electrode assemblies are arranged at intervals in a horizontal plane, the horizontal distance between the two groups of electrode assemblies is D, and the radius of curvature at any position of the axial lead of the connecting pipe section between the two groups of electrode assemblies is larger than D/4.
6. The flow electroporation assembly of claim 5, wherein: the radius of curvature of the axis at least part of the location of the connecting pipe section is substantially equal to D/2.
7. A flow electroporation component as claimed in claim 3, wherein: along the transport direction of the cell suspension: every two adjacent electrode assemblies are arranged at intervals along a horizontal first direction, the connecting pipe section between the two electrode assemblies is provided with a first rotary section, a second rotary section and a third rotary section, the first rotary section, the second rotary section and the third rotary section are sequentially connected from back to front along the transmission direction of the cell suspension, and are positioned on the same side of the second rotary section along the first direction.
8. The flow electroporation assembly of claim 7, wherein: along the transport direction of the cell suspension: in each two adjacent groups of electrode assemblies, the electrode assembly positioned at the front side is a front electrode assembly, the electrode assembly positioned at the rear side is a rear electrode assembly, the connecting pipe section between the front electrode assembly and the rear electrode assembly is provided with at least two second revolving sections, the second revolving section close to the front electrode assembly in the two second revolving sections is a front second revolving section, and the second revolving section close to the rear electrode assembly is a rear second revolving section;
along the first direction: the front second turnaround section is adjacent to the rear electrode assembly and the rear second turnaround section is adjacent to the front electrode assembly.
9. The flow electroporation assembly of claim 1, wherein: the connecting pipeline comprises a first pipe group and a second pipe group, the first pipe group comprises a first main pipeline and at least two first branches which are communicated with each other, the first main pipeline is connected with the main liquid inlet, and the electrode inlets of each group of electrode assemblies are correspondingly connected with one first branch;
the second tube group comprises a second main path and at least two second branches which are communicated with each other, the second main path is connected with the main liquid outlet, and each electrode outlet of each electrode assembly is correspondingly connected with one second branch.
10. The flow electroporation component of claim 9, wherein: the first pipe group is positioned below the second pipe group, the first main path is positioned below all the first branches, and the second main path is positioned above all the second branches.
11. The flow electroporation assembly of claim 1, wherein: the main liquid inlet is positioned below the main liquid outlet.
12. The flow electroporation component of claim 11, wherein: the axial lead of the main liquid inlet and the axial lead of the main liquid outlet extend along the up-down direction respectively, and the axial lead of the main liquid inlet and the axial lead of the main liquid outlet extend in a collinear way.
13. The flow electroporation assembly of claim 1, wherein: the electrode inlets of different electrode assemblies are located at the same height, and the electrode outlets of different electrode assemblies are located at the same height.
14. The flow electroporation assembly of claim 1, wherein in an up-down direction: the length of the electroporation chambers of all the electrode assemblies is equal, and the cross-sectional areas of the electroporation chambers of all the electrode assemblies at any positions are equal.
15. The flow electroporation assembly of claim 1, wherein: the electrode assemblies are arranged at intervals along the horizontal first direction, and the axial lead of the electroporation chambers of all the electrode assemblies and the axial lead of the connecting pipeline are positioned in the same plane; or alternatively, the first and second heat exchangers may be,
the axis of the main liquid inlet, the axis of the main liquid outlet, the axis of the connecting pipeline and the axes of all electroporation chambers of the electrode assemblies are all positioned in the same vertical plane.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211026908.0A CN117660176A (en) | 2022-08-25 | 2022-08-25 | Flow electroporation component |
| PCT/CN2023/104609 WO2024041200A1 (en) | 2022-08-25 | 2023-06-30 | Flow electroporation assembly |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211026908.0A CN117660176A (en) | 2022-08-25 | 2022-08-25 | Flow electroporation component |
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| CN117660176A true CN117660176A (en) | 2024-03-08 |
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| CN202211026908.0A Pending CN117660176A (en) | 2022-08-25 | 2022-08-25 | Flow electroporation component |
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| WO (1) | WO2024041200A1 (en) |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2523579A (en) * | 2014-02-28 | 2015-09-02 | Ales Belic | Modular serial multi-chamber flow-through electroporation device |
| WO2018200873A1 (en) * | 2017-04-26 | 2018-11-01 | Ravata Solutions, Inc. | Microelectrode techniques for electroporation |
| CN208250334U (en) * | 2017-10-19 | 2018-12-18 | 苏州壹达生物科技有限公司 | A kind of flow electroporation device |
| US11377652B2 (en) * | 2018-07-09 | 2022-07-05 | NanoCav, LLC | Micro flow-through electroporation devices and methods of cell transfection |
| CN110872559A (en) * | 2018-08-29 | 2020-03-10 | 苏州壹达生物科技有限公司 | Flow type electrotransfection device |
| JP2022554401A (en) * | 2019-11-11 | 2022-12-28 | プレシゲン，インコーポレイテッド | Electroporation apparatus and method |
| CN217948143U (en) * | 2022-08-25 | 2022-12-02 | 苏州壹达生物科技有限公司 | Flow type electroporation modularization device |
| CN217948142U (en) * | 2022-08-25 | 2022-12-02 | 苏州壹达生物科技有限公司 | Flow type electroporation device |
| CN217948141U (en) * | 2022-08-25 | 2022-12-02 | 苏州壹达生物科技有限公司 | Flow type electroporation component |
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- 2022-08-25 CN CN202211026908.0A patent/CN117660176A/en active Pending
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