KR102348640B1 - An electroporation apparatus - Google Patents

Abstract

The present invention relates to a chamber for an electroporation apparatus capable of perforating cells with high capacity and high efficiency.
The chamber for an electroporation apparatus according to an embodiment of the present invention includes a plate-shaped case having an enclosed space therein; a sample supply port communicating with the sealed space and provided outside the case; a sample outlet in communication with the sealed space and provided on the outside of the case; an air inlet communicating with the sealed space and provided on the outside of the case; an internal electrode provided in the sealed space and to which a high voltage is applied; and an external electrode electrically connected to the internal electrode and provided outside the case, wherein the case has a ring-shaped sealed space provided therein, and the internal electrode is disposed along an inner circumferential surface of the sealed space The provided anode electrode part and the cathode electrode part facing the anode electrode part and provided along the outer peripheral surface of the sealed space may be included.

Description

An electroporation apparatus

The present invention relates to an electroporation device capable of perforating cells with high capacity and high efficiency.

In general, electroporation is a gene introduction method in which a cell is placed in a DNA solution and a pulse of DC high voltage is passed through the DNA solution, creating a temporary hole in the cell membrane and introducing the DNA into the cell.

This electroporation method can be applied to various cell types, such as cells extracted from animals, plants, microorganisms, humans, etc., and RNA, siRNA, peptides, proteins, antibodies, drugs or other substances can be introduced into cells as well as DNA.

1 is a view showing an example of an electroporation apparatus according to the prior art.

As shown in FIG. 1, the conventional cell puncture device is provided with a first sample storage bag (1), a pump (2), a cell puncture chamber (3), and a second sample storage bag (4), connected The tube 5 is configured to sequentially connect each component.

When the pump 1 is operated, the sample before processing is supplied to the cell perforation chamber 3 through the pump 2 in the first sample storage bag 1, and when a high voltage is applied to the cell perforation chamber 3, the cell perforation The cell membrane of the cells is opened in the chamber 3 , an active material is introduced into the cells, and then the processed sample discharged from the cell perforation chamber 3 is stored in the second sample storage bag 4 .

According to the above technique, there is a problem in that the cell viability is lowered because the sample directly passes through the tube in the pump.

Korea Patent Publication No. 2017-0017876 (application on April 28, 2015) discloses a device for applying an electric field to a suspension of cells, cell derivatives, organelles, subcellular particles and/or vesicles, and the sample does not pass through the pump. This can increase cell viability.

The device comprises at least one chamber comprising at least two electrodes and at least one separation element, the separation element being movable within the chamber between the two endpoints, the separation element being in a position between the endpoints. case, the at least one first compartment of the chamber may be separated from the at least one second compartment of the chamber.

Each compartment is designed to hold a suspension and comprises at least one port for filling or discharging the suspension, thus discharging an aliquot of the suspension from the chamber and at the same time filling another aliquot of the suspension into the chamber, the separation element comprising the first Moved in a second direction opposite to the direction, the separating element separates the aliquots from each other.

According to the above technique, since the structure inside the chamber is complicated, it is difficult to process a large-capacity sample, and there is a problem in that the production cost is increased.

The present invention has been devised in order to solve the problems of the prior art, and an object of the present invention is to provide an electroporation apparatus capable of perforating cells with high capacity and high efficiency.

Electroporation apparatus according to an embodiment of the present invention, a chamber to which a voltage for cell perforation is applied; a supply bag in which the sample to be supplied to the chamber is stored; a recovery bag in which the sample that has passed through the chamber is recovered; a sample supply pipe connecting the supply bag and the chamber; a sample recovery tube connecting the chamber and the recovery bag; a pump that applies a negative pressure to the inside of the chamber, causes the sample stored in the supply bag to flow into the chamber, and applies a positive pressure to the inside of the chamber, so that the sample in the chamber is recovered into the recovery bag; and a pump connection pipe connecting the chamber and the pump, wherein the chamber includes: a case having an enclosed space filled with the sample supplied from the supply bag therein; a sample supply port communicating with the sealed space and provided outside the case to which the sample supply pipe is connected; a sample outlet communicating with the sealed space and provided outside the case to which the sample recovery pipe is connected; an air inlet communicating with the sealed space, provided outside the case, to which the pump connection pipe is connected; an internal electrode provided inside the sealed space and to which a high voltage is applied; and an external electrode electrically connected to the internal electrode and provided outside the case.

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The chamber for an electroporation device according to the present invention is provided with positive/negative electrodes on the inner/outer circumferential surface of a ring-shaped closed space through which a sample passes, or a plurality of positive/negative electrode electrodes disposed at regular intervals in the closed space. , it is possible to increase cell processing efficiency by allowing multiple electric fields to pass through a single cell included in a sample of the same volume.

Therefore, since the specification of the power supply unit providing the high voltage can be lowered, the production cost can be reduced and the size of the chamber can be reduced, so that the product can be miniaturized.

In addition, since the structure inside the chamber is simple and the sample can be filled in the sealed space inside the chamber, it is possible to process a large amount of sample, and there is an advantage in that it is possible to reduce the production cost.

1 is a view showing an example of an electroporation apparatus according to the prior art.
Figure 2 is a block diagram showing the control relationship of the electroporation apparatus according to the present invention.
Figure 3 is a block diagram showing a first embodiment of the electroporation apparatus according to the present invention.
Figures 4 to 5 are a perspective view and a side cross-sectional view showing the first embodiment of the chamber applied to Figure 3;
Figure 6 is a block diagram showing a second embodiment of the electroporation apparatus according to the present invention.
7 to 8 are a side view and a side cross-sectional view showing a second embodiment of the chamber applied to Fig. 6;
9 to 10 are plan sectional views showing a second embodiment of the chamber applied to FIG.
11 to 12 are perspective and side cross-sectional views showing a third embodiment of a chamber that can be applied to the electroporation apparatus according to the present invention.
13 to 14 are perspective and side cross-sectional views showing a fourth embodiment of a chamber that can be applied to the electroporation apparatus according to the present invention.

Hereinafter, the present embodiment will be described in detail with reference to the accompanying drawings.

Figure 2 is a block diagram showing the control relationship of the electroporation apparatus according to the present invention.

The control relationship of the electroporation apparatus according to the present invention may include a body unit 10, a power supply unit 20, and a control unit 30, as shown in FIG.

The body unit 10 may be equipped with an electroporation device to be described below, and an electrode output unit 10a capable of applying a high voltage current to the front surface is exposed in the form of a groove or a protrusion.

The power supply unit 20 applies a voltage to the main body unit 10 , and may provide a voltage of an appropriate level in which cell perforation may occur.

The higher the voltage, the higher the cell perforation efficiency, but the cell viability may decrease. Therefore, the power supply unit 20 preferably provides a voltage of an appropriate size to increase the cell perforation efficiency and the cell viability.

The control unit 30 may control the operation of the main body unit 10 and the power supply unit 20 .

For example, the control unit 30 may output a control parameter to the main unit 10 and may receive data from the main unit 10 , but is not limited thereto.

For example, the control unit 30 may output an input voltage to the power supply unit 20 , and may receive a voltage state from the power supply unit 20 , but is not limited thereto.

Figure 3 is a block diagram showing a first embodiment of the electroporation apparatus according to the present invention.

In the first embodiment of the electroporation apparatus according to the present invention, as shown in FIG. 3 , the circular back surface of the chamber 100 is the main body unit (10: shown in FIG. 2) in a state in which the disk-shaped chamber 100 is erected. As a form mounted on the front, it may include a supply bag (B1), a recovery bag (B2), and a pump (P).

The chamber 100 is electrically perforated while the sample passes, and the electrode connection part 100a protruding from the circular rear surface of the chamber 100 is formed in the front surface of the main body unit 10 (shown in FIG. 2). It may be mounted on the part 10a (shown in FIG. 2 ). A detailed configuration of the chamber 100 will be described below.

The supply bag B1 is in the form of a bag in which a sample to be supplied to the chamber 100 is stored, and may be replaceably mounted on the main body unit 10 (shown in FIG. 2 ). The supply bag B1 is connected to the lower portion of the chamber 100 by the sample supply pipe L1, and the supply valve V1 for controlling whether or not the sample is supplied may be provided in the sample supply pipe L1.

The recovery bag B2 is in the form of a bag in which the sample recovered from the chamber 100 is stored, and similarly, it may be replacedly mounted on the main body unit 10 (shown in FIG. 2 ). The recovery bag B2 is connected to the lower portion of the chamber 100 by a sample recovery pipe L2, and the sample recovery pipe 12 may be provided with a recovery valve V2 for controlling whether or not the sample is recovered.

The pump P is in the form of an air pump for regulating the pressure inside the chamber 100, and is provided with only one, but any metering pump capable of transferring fluid such as a tube peristaltic pump may be applied, and is not limited thereto.

The pump P is mounted on the main body unit 10 (shown in FIG. 2 ), and may be connected to the upper portion of the chamber 100 by a pump connection pipe L3 . When the above-described control unit 30 (shown in Fig. 2) transmits control parameters to the main body unit 10 (shown in Fig. 2), the operation of the pump P connected to the control unit 30 (shown in Fig. 2) By controlling the flow rate, flow rate, etc. of the sample passing through the chamber 100 can be controlled.

4 to 5 are perspective and side cross-sectional views illustrating a first embodiment of the chamber applied to FIG. 3 .

The chamber 100 according to the first embodiment of the present invention includes a case 110 in which a ring-shaped sealed space (S) is formed, as shown in FIGS. 4 to 5 , and inner/outer peripheral surfaces of the sealed space (S). It may include an internal electrode 120 mounted accordingly, and an external electrode 130 exposed to the outside of the case 110 so as to be electrically connected to the internal electrode 120 .

The case 110 has a ring-shaped sealed space S formed therein, and the case 110 is provided with a sample inlet 111 and a sample outlet 112 for injecting and discharging a sample into the sealed space S. and an air inlet 113 for discharging air from the closed space S may be provided.

The sample inlet 111 and the sample outlet 112 are provided side by side on the lower front side of the case 110, and the sample inlet 111 is provided in a supply bag (B1: in FIG. 2) by a sample supply pipe (L1: shown in FIG. 2). shown), and the sample outlet 112 may be connected to the recovery bag B2 (shown in FIG. 2 ) by a sample discharge pipe L2 (shown in FIG. 2 ). The air inlet 113 is provided on the front upper portion of the case 110, and may be connected to the pump (P: shown in FIG. 2) by a pump connection pipe (L3: shown in FIG. 2).

The sample inlet 111 , the sample outlet 112 , and the air inlet 113 may be provided on one or both sides of the case 110 , or any one of the outer peripheral surfaces, in consideration of the location where the sample is easily transported and air is discharged. The design may be appropriately changed.

In order to prevent leakage of the sample injected into the sealed space (S), the case 110 itself is composed of a sealing member 114 having a predetermined elasticity, or the sealing member 114 is built-in in the case 110 . can be composed of

Since the internal electrode 120 is provided inside the sealing member 114 , the sealing member 114 must be made of an insulating material to insulate the internal electrode 120 .

Accordingly, the sealing member 114 may be made of an insulating material having a predetermined elasticity that is harmless to the human body, and may be typically made of medical silicone, but is not limited thereto.

The internal electrode 120 is provided so as to flow a high voltage to the sample, and the anode electrode part 121 provided along the inner circumferential surface of the sealed space S, and the sealed space S to face the anode electrode part 121 . It may include a negative electrode part 122 provided along the outer peripheral surface of the.

In order to apply a uniform voltage to the entire sample passing through the enclosed space S, the positive electrode part 121 and the negative electrode part 122 may be disposed to be spaced apart from each other by a predetermined distance in the radial direction.

The anode electrode part 121 and the cathode electrode part 122 may be made of any one of various metals, such as copper, aluminum, SUS, etc. to which a high voltage may be applied, but is not limited thereto.

Since the positive electrode part 121 and the negative electrode part 122 come into contact with the sample containing cells, it is preferable to be plated with any one of high-conductivity metals harmless to the human body, and may be plated with gold, platinum, etc., but is not limited thereto. No.

The external electrode 130 may include a positive terminal connected to the positive electrode part 121 and a negative terminal connected to the negative electrode part 122 .

The external electrode 130 may be variously configured with metal parts such as a banana plug and a fastening screw, and may be connected to the electrode output unit 10a (shown in FIG. 2 ) of the body unit.

According to the embodiment, the external electrode 130 is in the form of a fastening screw, and may be composed of a screw thread part 131 and a head part 132 , and the center of the rear surface of the case 110 so that the external electrode 130 can be fastened. A fastening hole 110h may be provided in the . The screw thread part 131 may protrude axially outside the fastening hole 110h, and the head 132 may be supported inside the fastening hole 110h and connected to the internal electrode 120 at the same time.

By configuring the external electrode 130 as described above, one of the internal electrodes 120 can be connected to the electrode output part 10a of the body unit (shown in FIG. 2 ), and a separate groove (not shown) is provided in the case 110 . ) to expose the remaining electrode parts of the internal electrodes 120 to the outside, and the electrode part exposed to the outside of the case 110 may be separately connected to the electrode output part 10a of the body unit (shown in FIG. 2 ).

Looking at the operating state of the electroporation apparatus according to the first embodiment to which the chamber according to the first embodiment configured as described above is applied with reference to Figs. 3 to 5, as follows.

When the supply valve (V1) is opened and the pump (P) is operated to apply a negative pressure to the inside of the chamber 100, the air inside the chamber 100 is discharged to the outside of the chamber 100 through the pump connection pipe (L3) As the pressure inside the chamber 100 is lowered, the sample before processing contained in the supply bag B1 is injected into the chamber 100 through the sample supply pipe L1, and the sample before processing is placed in a ring-shaped closed space ( S) Filled from bottom to top.

When the sample fills the sealed space S, an appropriate high voltage is temporarily applied to the positive electrode part 121 and the negative electrode part 122 , and is located between the positive electrode part 121 and the negative electrode part 122 . A high voltage also flows through the sample. Accordingly, the membrane of cells included in the sample is temporarily opened by the high voltage, and the cell perforation process in which active substances included in the sample are introduced into the cells may proceed.

When the cell perforation process is completed, the recovery valve V2 is opened, and when the pump P is operated to apply a positive pressure to the inside of the chamber 100 , air outside the chamber 100 flows into the chamber through the pump connection pipe L3 . (100) flows into the interior, and as the pressure inside the chamber 100 increases, the sample inside the chamber 100 is recovered to the recovery bag B2 through the sample discharge pipe L2, and the processed sample is returned to the recovery bag ( It can be stored in B2).

In this way, when the cell puncture process is performed in the chamber 100 provided in the inner/outer peripheral surfaces of the ring-shaped closed space S in the form of the anode electrode part 121 and the cathode electrode part 122 in a curved shape, the anode electrode part and Compared to a chamber in which the cathode electrode part is spaced side by side in a planar shape, multiple electric fields can pass through a single cell within the same volume, and cell perforation efficiency can be increased.

6 is a block diagram showing a second embodiment of the electroporation apparatus according to the present invention.

In the second embodiment of the electroporation apparatus according to the present invention, as shown in FIG. 6, one side of the outer periphery of the chamber 200 is the main body unit (10: shown in FIG. 2) in a state in which the disk-shaped chamber 200 is erected. As a form mounted on the front of the, it may include a supply bag (B1), a recovery bag (B2), and a pump (P).

The configuration of the chamber 200 will be described in detail below, and the remaining components are configured the same as those of the first embodiment, so a detailed description will be omitted.

The electroporation apparatus according to the second embodiment of the present invention can mount the chamber 200 to the main body unit (10: shown in FIG. 2) in a slim form compared to the first embodiment, and an operator can install the chamber 200 The grip feeling can be improved when holding and attaching to the main body unit 10 (shown in FIG. 2 ).

7 to 8 are side views and cross-sectional views showing a second embodiment of the chamber applied to FIG. 6 , and FIGS. 9 to 10 are plan sectional views showing a second embodiment of the chamber applied to FIG. 6 .

The chamber 200 according to the second embodiment of the present invention includes a case 210 in which a ring-shaped sealed space S is formed, as shown in FIGS. 7 to 10 , and inner/outer peripheral surfaces of the sealed space S. It may include an internal electrode 220 mounted accordingly, and an external electrode 230 exposed to the outside of the case 210 so as to be electrically connected to the internal electrode 220 .

The case 210 may be composed of a pair of covers 210a and 210b that form a ring-shaped sealed space S as they are assembled with each other in the axial direction, and the covers 210a and 210b are PC, ABS, etc. It can be made of one of a variety of materials that are easy to use.

The covers (210a, 210b) may be configured in a stepped shape so that the portions in contact with each other are formed, and a plurality of fastening holes (h) are formed in the protruding portions at regular intervals on the inner and outer circumferences of the covers (210a, 210b). ) may be provided, and as the bolts b are fastened to the fastening holes h of the covers 210a and 210b, the covers 210a and 210b may be assembled with each other.

The case 210 may be provided with a sample inlet 211 , a sample outlet 212 , and an air inlet 213 to communicate with the sealed space S, and is appropriately design may be changed.

According to an embodiment, the sample inlet 211 and the sample outlet 212 may be provided in a line on the lower front/rear surface of the case 210 , and the air inlet 213 may be provided on the upper front surface of the case 210 . can, but is not limited to.

However, when the case 210 is composed of a pair of covers 210a and 210b, since the sample may leak between the covers, the sealing member 214 may be built in the case 210 and the sealing member ( 214) may be provided in the closed space (S) on the inside.

The sealing member 214 in direct contact with the sample may be made of an insulating material having a predetermined elasticity that is harmless to the human body, and may be typically made of medical silicone, but is not limited thereto.

The sample inlet 211, the sample outlet 212, and the air inlet 213 may be integrally formed with the sealing member 214 in a tube type, and the sample inlet 211, the sample outlet 212 and the air inlet 211 of the tube type The doorway 213 may be exposed outside the covers 210a and 210b.

The internal electrode 220 includes an anode electrode part 221 and a cathode electrode part 222 , and is the same as the internal electrode applied to the chamber of the first embodiment, and thus a detailed description thereof will be omitted.

The external electrode 230 is provided outside the case 210 to be electrically connected to the internal electrode 220 , and may include a positive terminal 231 and a negative terminal 232 .

The positive electrode terminal 231 may be integrally formed with the positive electrode part 221 , and may be configured in the form of a screw thread protruding in the inner radial direction of the positive electrode part 221 .

The negative electrode terminal 232 may be integrally formed with the negative electrode part 222 , and may be configured in the form of a screw thread protruding radially outside the negative electrode part 222 .

A pair of fixing nuts 233 and 234 may be provided to respectively fix the positive/negative terminals 231,232 to the inner/outer peripheral surfaces of the case 210 . When each of the fixing nuts 233 and 234 is assembled along the positive/negative terminals 231,232, they may be seated on the inner/outer peripheral surfaces of the case 210, and the positive/negative terminals 231,232 may be fixed to the case 210. .

The positive/negative terminals 231,232 may be connected to the electrode output unit 10a (shown in FIG. 2) of the body unit, and the negative terminal 232 is directly connected to the electrode output unit 10a (shown in FIG. 2) of the body unit. can

Since the operation of the chamber according to the second embodiment configured as described above is the same as that of the chamber of the first embodiment, a detailed description thereof will be omitted.

11 to 12 are perspective and side cross-sectional views showing a third embodiment of a chamber that can be applied to the electroporation apparatus according to the present invention.

The chamber 300 according to the third embodiment of the present invention includes a case 310 having a closed space (S) in the shape of a flattened square plate as shown in FIGS. 11 to 12 , and a closed space (S). It may include a plurality of electrode rod-shaped internal electrodes 320 provided to cross the vertical direction, and external electrodes 330 exposed outside the case 310 to be electrically connected to the internal electrodes 320 .

The case 310 may include a positive electrode holder 311 and a negative electrode holder 312 spaced apart from each other in a vertical direction as a form in which the closed space S in the shape of a square plate can be mounted in a lying state.

The positive electrode holder 311 has a rectangular plate shape made of an insulating material having a predetermined elasticity, and may be provided with a plurality of holes 311h through which the upper portion of the electrode can be inserted at a predetermined interval.

The negative electrode holder 312 is also in the shape of a square plate having the same material and size as the positive electrode holder 311 , and may be provided with a plurality of holes 312h through which the lower portion of the electrode can be inserted at regular intervals.

When the positive electrode holder 311 and the negative electrode holder 312 are assembled to face each other in the vertical direction, the holes 311h of the positive electrode holder and the holes 312h of the negative electrode holder are arranged so as not to overlap in the vertical direction, and the holes of the positive electrode holder ( 311h), holes 312h of the cathode holder may be disposed.

The case 310 may further include a cover 313 for assembling the anode holder 311 and the cathode holder 312 , and the cover 313 may be made of an insulating material having a predetermined elasticity. The cover 313 is assembled in a state where the positive electrode holder 311 and the negative electrode holder 312 are vertically spaced apart, and may be installed to surround the outer peripheral surfaces of the positive electrode holder 311 and the negative electrode holder 312 .

Although not shown in the drawings, a sample inlet, a sample outlet, and an air inlet communicating with the closed space S may be provided in the case 310, and the location thereof is not limited.

The internal electrode 320 may include a plurality of positive electrode rods 321 and a plurality of negative electrode rods 322, and the lengths of the positive electrode electrode 321 and the negative electrode electrode 322 may be the same or irregularly configured. , but not limited.

Of course, the anode electrode 321 and the cathode electrode 322 are materials that can be subjected to high voltage, and are preferably plated with one of high-conductivity metals that are harmless to the human body even when in contact with the sample, and may be plated with gold, platinum, etc. , but not limited.

The upper portions of the positive electrode rods 321 are fitted into the holes 311h of the positive electrode holder, the lower portions of the negative electrode rods 322 are inserted into the holes 312h of the negative electrode holder, and then the positive electrode holder 311 and the negative electrode holder 312 ) is assembled by the cover 313 , the lower portions of the positive electrode rods 321 are supported on the negative electrode holder 312 , and the upper portions of the negative electrode rods 322 are supported on the positive electrode holder 311 , the positive electrode rods 321 and the cathode electrodes 322 may be alternately disposed adjacent to each other in the enclosed space (S).

The external electrode 330 may be composed of an anode electrode plate 331 and a cathode electrode plate 332 , and may be made of an aluminum material capable of applying a high voltage, but is not limited thereto.

The positive electrode plate 331 has a size capable of covering the top surface of the positive electrode holder 311 , and is disposed on the upper side of the positive electrode holder 311 , so that it can be mounted to be electrically connectable to the top of the positive electrode rods 321 . .

The negative electrode plate 332 is sized to cover the lower surface of the negative electrode holder 312 , and is disposed below the negative electrode holder 312 , so that the negative electrode plate 332 may be electrically connected to the lower ends of the negative electrode rods 322 . .

Although not shown in the drawings, the positive electrode plate 331 and the negative electrode plate 332 are shielded with a separate insulating material that engages the upper and lower portions of the cover 313 , and the positive electrode plate 331 and the negative electrode plate 331 . Only a part of the plate 332 is configured to be exposed outside the shielding structure, and the exposed part of the positive electrode plate 331 and the negative electrode plate 332 can be connected to the power output unit 10a of the main body unit (shown in FIG. 2 ) have.

The chamber 300 according to the third embodiment configured as described above arranges the positive electrode electrodes 321 and the negative electrode rods 322 adjacent to each other in the enclosed space S, so that the interior of the enclosed space S is uniformly formed. An electric field can be formed, and a contact area between the sample and the electrodes 321 and 322 can be increased, thereby maximizing the electroporation efficiency.

13 to 14 are perspective and side cross-sectional views showing a fourth embodiment of a chamber that can be applied to the electroporation apparatus according to the present invention.

The chamber 400 according to the fourth embodiment of the present invention includes a case 410 in which a closed space (S) of a rhombus shape erected as shown in FIGS. 13 to 14 is formed, and the closed space (S) in the vertical direction. It may include a plurality of electrode rod-shaped internal electrodes 420 provided to cross, and external electrodes 430 exposed outside the case 410 to be electrically connected to the internal electrodes 420 .

The case 410 is a form that can be mounted in a state in which the closed space (S) of the rhombus shape is erected, and the volume of the closed space (S) increases and then decreases from the lower part to the upper part.

The case 410 may include a pair of covers 411 and 412 spaced apart in both directions in a rhombus shape, and a holder 413 provided along a circumferential portion between the covers 411 and 412 .

The holder 413 may be provided with holes 413h and 413h' into which the upper or lower portions of the electrode rods 420 can be fitted. The lower holes 413h' of the holder may be positioned between the upper holes 413h of the holder, instead of being positioned at positions overlapping each other.

The internal electrode 420 may include a plurality of positive electrode electrodes 421 and a plurality of negative electrode electrodes 422 disposed adjacent to each other inside the closed space S.

Upper portions of the anode electrodes 421 are fitted into the upper holes 413h of the holder 413 , and lower portions of the cathode electrodes 422 are fitted into the lower holes 413h′ of the holder 413 , and then a pair of When the covers 411 and 412 are assembled on both sides of the holder 413 , the lower portions of the positive electrode rods 421 are supported on the lower portion of the holder 413 , and the upper portions of the negative electrode rods 422 are supported on the upper portion of the holder 413 . In addition, the positive electrode electrodes 421 and the negative electrode rods 422 may be alternately disposed adjacent to each other in the enclosed space (S).

The external electrode 430 may include a positive electrode plate 431 and a negative electrode plate 432 made of a material capable of applying a high voltage.

The positive electrode plate 431 may be provided outside the upper portion of the holder 413 so as to be in contact with the upper ends of the positive electrode rods 421 , and portions other than a portion may be shielded by an insulating material.

The negative electrode plate 432 may be provided outside the lower portion of the holder 413 so as to be in contact with the upper ends of the negative electrode electrodes 422 , and portions other than a portion may be shielded by an insulating material.

The chamber 400 according to the fourth embodiment configured as described above can be mounted in an electroporation apparatus like the chamber of the second embodiment, and forms a uniform electric field with a multi-electrode structure as in the chamber of the third embodiment and at the same time electroporation efficiency can be maximized.

The above description is merely illustrative of the technical spirit of the present invention, and various modifications and variations will be possible without departing from the essential characteristics of the present invention by those skilled in the art to which the present invention pertains.

Therefore, the embodiments disclosed in the present invention are not intended to limit the technical spirit of the present invention, but to explain, and the scope of the technical spirit of the present invention is not limited by these embodiments.

The protection scope of the present invention should be construed by the following claims, and all technical ideas within the scope equivalent thereto should be construed as being included in the scope of the present invention.

100: chamber 110: case
120: inner electrode 130: outer electrode

Claims (20)

a energized chamber for cell perforation;
a supply bag in which the sample to be supplied to the chamber is stored;
a recovery bag in which the sample that has passed through the chamber is recovered;
a sample supply pipe connecting the supply bag and the chamber;
a sample recovery tube connecting the chamber and the recovery bag;
a pump that applies a negative pressure to the inside of the chamber, causes the sample stored in the supply bag to flow into the chamber, and applies a positive pressure to the inside of the chamber, so that the sample in the chamber is recovered into the recovery bag; and
A pump connection pipe connecting the chamber and the pump,
The chamber is
a case having an enclosed space filled with the sample supplied from the supply bag;
a sample supply port communicating with the sealed space and provided outside the case to which the sample supply pipe is connected;
a sample outlet communicating with the sealed space and provided outside the case to which the sample recovery pipe is connected;
an air inlet communicating with the sealed space, provided outside the case, to which the pump connection pipe is connected;
an internal electrode provided inside the sealed space and to which a high voltage is applied; and
Electroporation apparatus comprising a; electrically connected to the inner electrode, the external electrode provided on the outside of the case. The method of claim 1,
The sealed space is formed in a ring shape inside the case,
The inner electrode is
a positive electrode part provided along the inner circumferential surface of the sealed space;
An electroporation apparatus facing the anode electrode part and including a cathode electrode part provided along an outer circumferential surface of the sealed space. 3. The method of claim 2,
Electroporation apparatus further comprising a sealing member for filling the inner space of the case except for the sealed space. 4. The method of claim 3,
The case is
It consists of a shape symmetrical to each other, and includes a pair of covers that are coupled to each other to accommodate the closed space,
Electroporation apparatus, characterized in that the pair of covers are combined to form a ring-shaped chamber. 5. The method of claim 4,
The sealing member,
An electric drilling device including a pair of sealing members provided inside the case and having a ring shape symmetrical to each other. 6. The method of claim 5,
Any one of the pair of sealing members formed in a ring shape shields an opening formed at one end of the electrode part to define any one of an upper surface and a lower surface of the sealed space,
The other one of the pair of sealing members formed in a ring shape shields the opening formed at the other end of the electrode part to define the other one of the upper surface and the lower surface of the sealed space,
An electric drilling apparatus, characterized in that the sealed space is formed in a ring shape by the electrode part and the sealing member. 4. The method of claim 3,
The sealing member is an electric drilling device composed of an insulating material having a predetermined elasticity. 3. The method of claim 2,
The positive electrode part and the negative electrode part,
Electroporation device plated with one of the highly conductive metals harmless to the human body. 3. The method of claim 2,
In this case,
Electroporation device provided with a groove for exposing a portion of the anode electrode portion to the outside of the case 3. The method of claim 2,
The external electrode is
a positive terminal connected to the positive electrode part and protruding radially inward of the case;
Electroporation device connected to the negative electrode part and including a negative terminal protruding outward in the radial direction of the case 11. The method of claim 10,
At least one of the positive terminal and the negative terminal is an electroporation device provided in the shape of an electrode protruding from the case. The method of claim 1,
The sample supply port,
It is formed on one side of the lower part of the closed space,
The sample outlet is
An electric drilling device formed on the other side of the lower portion of the closed space. The method of claim 1,
The sample inlet and the sample outlet are
Electroporation device arranged in a direction parallel to the central axis of the case. The method of claim 1,
The case is
A rhombus-shaped positive electrode holder and negative electrode holder that are horizontally spaced apart from each other at positions opposite to each other;
a side cover surrounding the edges of the positive electrode holder and the negative electrode holder;
a positive electrode plate covering the outer surface of the positive electrode holder;
and a negative electrode plate covering the outer surface of the negative electrode holder. 15. The method of claim 14,
Inside the case,
A closed space in the shape of a square plate is formed,
The inner electrode is
a plurality of positive electrode rods extending from the positive electrode holder toward the negative electrode holder across the sealed space;
a plurality of negative electrode rods extending from the negative electrode holder toward the positive electrode holder across the sealed space;
The plurality of positive electrode electrodes and the negative electrode electrode are alternately spaced apart from one side of the side cover to the other side,
Electroporation apparatus, characterized in that the holes into which the ends of the positive electrode and the negative electrode are inserted are respectively formed on the opposite surfaces of the positive electrode holder and the negative electrode holder. 15. The method of claim 14,
The side cover is
An electroporation device made of an insulating material having a predetermined elasticity. 16. The method of claim 15,
The positive electrode and the negative electrode are
Electroporation device plated with one of the high-conductivity metals harmless to the human body. 15. The method of claim 14,
The case is
A pair of rhombus-shaped covers that are vertically spaced apart from each other at positions opposite to each other;
And a holder that surrounds the edge of the pair of covers,
a plurality of negative electrode electrodes extending vertically from the upper portion of the holder;
A plurality of positive electrode electrodes extending vertically from the lower portion of the holder,
The plurality of negative electrode rods and the plurality of positive electrode rods are electric drilling apparatus, characterized in that the arrangement is alternately spaced apart in a direction from one cover to the other cover of the pair of covers. 19. The method of claim 18,
The case is
An electroporation device disposed in a shape where the cross-sectional area of the closed space gradually increases from the lower part to the upper part and then gradually decreases. 19. The method of claim 18,
The external electrode is
a positive electrode plate provided outside the positive electrode holder and electrically connected to one end of the positive electrode rods;
Electroporation apparatus comprising a negative electrode plate provided on the outside of the negative electrode holder and electrically connected to one end of the negative electrode rods.

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