Magnetic Bead Removal Method, Device, and Storage Medium

Abstract

A magnetic bead removal method, device, and storage medium are provided. The method includes the steps of placing a liquid bag having a curved flow path on a magnetic platform, wherein a plane where the curved flow path is located is positioned above the magnetic platform and parallel to the magnetic platform; controlling a first flow rate control assembly to communicate the liquid bag with a sample container, thereby injecting a liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path; flattening the liquid bag to increase a contact surface between a bottom surface of the curved flow path and the magnetic platform, and adsorbing immunomagnetic bead cells in the curved flow path onto the contact surface by the magnetic platform; and performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container.

Description

Technical Field

[0001] This application claims the priority of a Chinese patent application filed with the China National Intellectual Property Administration on September 27, 2022, with an application number of 202211181849.4 and an invention title of "Magnetic Bead Removal Method, Device, and Storage Medium", the entire content of which is incorporated herein by reference. This application relates to the technical field of cell separation, and particularly to a magnetic bead removal method, device, and storage medium.

Background Art

[0002] In the field of biotechnology, it is common to select specific cells or bacteria with magnetic beads and culture them for experiments or treatments. Currently, even after using micron-scale magnetic beads to select immunomagnetic bead cells and removing non-magnetic immune cells by culturing immunomagnetic bead cells, the immunomagnetic bead cells still remain in the culture medium. In order to avoid adverse effects on experiments or medical treatment in the next step, it may be necessary to adsorb and remove the magnetic beads by a magnetic field. The inventors of the present invention have found that in the prior art, in the process of removing immunomagnetic bead cells, due to the non-shear characteristics of the fluid, the area of the fluid bag containing the culture medium is too large (with the development of the cell processing industry, the volume of samples in the liquid bag of the culture medium is increasing, such as sample volumes of 5L and 10L), and the flow rate of the liquid bag containing the culture medium flowing in the magnetic field is extremely uneven (see Figure 7). As a result, in some areas where the flow rate is too fast, some immunomagnetic bead cells escape, and as a result, the immunomagnetic bead cells are not completely adsorbed by the magnetic field, resulting in a decrease in the utilization rate of the magnetic field. To address the above problems, if the escape of immunomagnetic bead cells is reduced by overall slowing down the flow rate of the cell fluid, this solution has drawbacks such as a long magnetization time.

Summary of the Invention

Problems to be Solved by the Invention

[0003] Embodiments of the present application provide a magnetic bead removal method, device, and storage medium to solve problems in the prior art such as the escape of immunomagnetic bead cells and low magnetic field utilization rate.

[0004] A magnetic bead removal method, comprising: placing a liquid bag having a curved flow path on a magnetic platform, wherein a plane where the curved flow path is located is above the magnetic platform and parallel to the magnetic platform; controlling a first flow rate control assembly to communicate the liquid bag with a sample container, thereby injecting a liquid containing magnetically bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path; flattening the liquid bag to increase a contact surface between a bottom surface of the curved flow path and the magnetic platform, and adsorbing immunomagnetic bead cells in the curved flow path onto the contact surface by the magnetic platform; performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container.

[0005] A magnetic bead removal device, comprising a pressing device and a controller for performing the above magnetic bead removal method, wherein the controller is connected to the pressing device.

[0006] A computer-readable storage medium storing computer-readable instructions that, when executed by a processor, implement the above magnetic bead removal method.

Advantages of the Invention

[0007] In the above magnetic bead removal method, device, and storage medium, the method includes the steps of placing a liquid bag having a curved flow path on a magnetic platform, wherein the plane where the curved flow path is located is positioned above the magnetic platform and parallel to the magnetic platform; controlling a first flow rate control assembly to communicate the liquid bag with a sample container, thereby injecting a liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path; pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved flow path and the magnetic platform, so that the magnetic platform adsorbs the immunomagnetic bead cells in the curved flow path onto the contact surface; and performing a collection operation to collect the non-magnetic immune cells in the curved flow path into a collection container. In the present application, by designing a curved flow path, the flow rates of the liquids in the liquid bag are made as consistent as possible, significantly improving the flow rate uniformity, sufficiently improving the adsorption ability of the magnetic platform for immunomagnetic bead cells in a uniform flow field, and improving the utilization rate of the magnetic field. Further, by pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved flow path and the magnetic platform, the magnetic platform can more sufficiently adsorb the immunomagnetic bead cells in the curved flow path with the increased contact surface, thereby avoiding the escape of immunomagnetic bead cells, improving the utilization rate of the magnetic field, ensuring the simplicity of the operation, and enabling the efficient and stable capture of immunomagnetic bead cells.

[0008] Details of one or more embodiments of the present application are shown in the following drawings and description, and other features and advantages of the present application will become apparent from the specification, drawings, and claims.

Brief Description of the Drawings

[0009] To more clearly explain the technical solution of the embodiments of this application, the drawings necessary for the description of the embodiments of this application will be briefly described below. However, the drawings in the following description are only some embodiments of this application, and it is obvious that those skilled in the art can obtain other drawings based on these drawings without creative efforts.

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Modes for Carrying Out the Invention

[0010] Hereinafter, with reference to the drawings in the embodiments of this application, the technical solutions in the embodiments of this application will be clearly and completely described. However, it is obvious that the described embodiments are only part of the embodiments of this application, not all of them. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative labor belong to the protection scope of this application.

[0011] In one embodiment, as shown in FIG. 1, a magnetic bead removal method including the following steps S10 to S40 is provided. S10: Place the liquid bag 60 having the curved flow path 61 on the magnetic platform 10. The plane where the curved flow path 61 exists is located above the magnetic platform 10 and is parallel to the magnetic platform 10. Here, the magnetic platform 10 may be a magnetic platform made of an electromagnet or a permanent magnet. For example, the magnetic platform 10 shown in FIG. 5 may include a base 101 provided with a lift through hole 1011 and a permanent magnet magnetic block 102 or an electromagnet attached to the base 101. An attachment groove is provided in the base 101, and the magnetic block 102 is fixedly attached to the attachment groove. As can be understood, the liquid bag 60 having the curved flow path 61 is made of a non-magnetic material and is not adsorbed by magnetic force, so it is not affected by the magnetic field of the magnetic platform 10 below it.

[0012] S20: By controlling a first flow rate control assembly (not shown) so as to communicate the liquid bag 60 with a sample container (not shown), inject the liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path 61. Here, the first flow rate control assembly may include a peristaltic pump capable of providing power for the liquid to flow and a pinch valve for opening and closing the first flow rate control assembly, and may also include a flow rate adjustment valve for controlling the flow rate of the liquid, a sensor for monitoring the flow velocity and pressure of the liquid, etc. The sample container contains the culture solution after culturing. The culture solution (i.e., the liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the above sample container) contains immune magnetic bead cells and non-magnetic immune cells without magnetic beads that have been incubated and divided in the medium by the immune magnetic bead cells (during culturing, the magnetic beads do not divide, but the cells do divide, so non-magnetic immune cells with antibodies but without magnetic beads can be obtained by division).

[0013] Furthermore, the liquid bag 60 includes a liquid inlet 62 communicating with the sample container and a liquid outlet 63 communicating with the collection container by the first flow rate control assembly, and the curved flow path 61 communicates between the liquid inlet 62 and the liquid outlet 63. In this embodiment, by the first flow rate control assembly, the culture solution in the sample container is sent from the liquid inlet 62 to the curved flow path 61, and further, by the action of the magnetic platform 10, the immunomagnetic bead cells can be adsorbed by the magnetic field generated by the magnetic platform 10. Non-magnetic immune cells other than the immunomagnetic bead cells flow from the liquid outlet 63 of the curved flow path 61 to the collection container. In this embodiment, since the material of the pipe connected to the curved flow path 61 is a non-magnetic material such as PVC material, it is not affected by the magnetic field of the magnetic platform 10.

[0014] In one embodiment, the curved flow path 61 includes a plurality of straight pipe segments 611 arranged in parallel, and connection pipe segments 612 connected between two adjacent straight pipe segments 611. The connection pipe segment 612 is a curved pipe or a straight pipe forming a preset inclination angle with the straight pipe segment 611 (the preset inclination angle is determined as needed. For example, the preset inclination angle shown in FIG. 4 is 90 degrees, that is, the straight pipe segment 611 and the connection pipe segment 612 are perpendicular to each other.), and / or the curved flow path 61 includes a spiral pipe segment. That is, in this embodiment, the curved flow path 61 may be a spiral pipe segment shaped like a mosquito coil incense, or a combination of the straight pipe segment 611 and the connection pipe segment 612 shown in FIG. 3 (the connection pipe segment 612 in FIG. 3 is a curved pipe) and FIG. 4 (the connection pipe segment 612 in FIG. 4 is a straight pipe). Further, in some embodiments, the curved flow path 61 may be formed by combining the straight pipe segment 611, the connection pipe segment 612, and the spiral pipe segment. It can be understood that in the present application, all pipe segments of the curved flow path are in the same plane, and this plane is parallel to the magnetic platform 10. In the above configuration, the curved flow path 61 is affected by the boundary layer, and the flow velocity of the liquid therein is relatively uniform, further increasing the magnetic field utilization rate of the magnetic platform 10. It can be understood that the shape of the curved pipeline may be set to other shapes as needed, as long as it can make the liquid flow velocity uniform and improve the magnetic field utilization rate. Further, the maximum width of the curved flow path 61 is 50 mm or less, and the curved flow path 61 with a width within this range has a high effect of improving the magnetic field utilization rate.

[0015] Furthermore, the liquid bag 60 further includes a buffer space, and the curved flow path 61 communicates with the liquid outlet 63 via the buffer space. That is, the buffer space is provided to buffer the liquid during the process of flowing through the curved pipeline and reaching the liquid outlet 63. Also, if necessary, it can change the flow direction of the liquid and slow down the flow rate. In one specific embodiment, the curved flow path 61 is a spiral tube segment shaped like a mosquito coil formed by coiling a 2×mm liquid tube (i.e., an inner diameter of 2 mm, an outer diameter of mm, and the liquid tube may have other sizes). A circular buffer space is provided at the center of the spiral tube segment, and the spiral tube segment communicates with the liquid outlet 63 via the buffer space.

[0016] S30: Press the liquid bag 60 to increase the contact surface between the bottom surface of the curved flow path 61 and the magnetic platform 10, and adsorb the immunomagnetic bead cells in the curved flow path 61 onto the contact surface by the magnetic platform 10. That is, the contact surface formed by the close contact of the liquid bag 60 and the magnetic platform 10 becomes larger. In this way, the magnetic platform 10 closely contacts the increased contact surface and the immunomagnetic bead cells in the liquid (culture solution) in the liquid bag 60 at a short distance, uniformly contacts the liquid with the magnetic field of the magnetic platform 10, and adsorbs the immunomagnetic bead cells onto the increased contact surface, so as to more fully perform magnetic adsorption in the magnetic separation process and avoid the immunomagnetic bead cells from not being adsorbed and flowing out of the liquid bag 60 through the liquid outlet 63 together with the non-magnetic immune cells. Also, after the liquid bag 60 is pressed flat, since the contact surface increases, it can achieve the effect of magnetizing and adsorbing the magnetic beads in a large volume of cell fluid all at once in the magnetic separation process. Thereby, magnetization can be performed all at once without controlling the flow rate, achieving the effect of achieving the consistency of magnetization.

[0017] S40: Perform a collection operation to collect the non-magnetic immune cells in the curved flow path 61 into a collection container (not shown). Further, in step S40, a second flow rate control assembly (not shown) provided between the collection container and the liquid bag 60 is controlled and activated to communicate the liquid bag 60 with the collection container, and the non-magnetic immune cells in the curved flow path 61 are transported to the collection container by the second flow rate control assembly. In this way, the non-magnetic immune cells are collected and used for experiments or medical treatment. That is, in this embodiment, the sample container and the collection container are both communicated with the liquid bag 60. The first flow rate control assembly is provided between the sample container and the liquid bag 60, and the second flow rate control assembly is provided between the collection container and the liquid bag 60. The second flow rate control assembly may include a peristaltic pump that provides power for the liquid to flow and a pinch valve for opening and closing the second flow rate control assembly, and may also include a flow rate adjustment valve for controlling the flow rate of the liquid, a sensor for monitoring the flow velocity and pressure of the liquid, etc. In the present application, when only one of the first flow rate control assembly and the second flow rate control assembly is provided, the flow parameters of the liquid in the liquid bag 60, such as velocity, flow rate, pressure, etc., can be controlled by the first flow rate control assembly or / and the second flow rate control assembly. Also, both the first flow rate control assembly and the second flow rate control assembly may be provided.

[0018] In the present application, by designing the curved flow path 61, the flow velocities of the liquid in the liquid bag 60 are made as consistent as possible, the uniformity of the flow velocity is greatly improved, the adsorption ability of the magnetic platform 10 for the immunomagnetic bead cells in a uniform flow field is sufficiently improved, and the utilization rate of the magnetic field is improved. Also, by pressing the liquid bag 60 flat to increase the contact surface between the bottom surface of the curved flow path 61 and the magnetic platform 10, the magnetic platform 10 can more sufficiently adsorb the immunomagnetic bead cells in the curved flow path 61 with the increased contact surface, thereby avoiding the escape of the immunomagnetic bead cells, improving the utilization rate of the magnetic field, ensuring the simplicity of the operation, and enabling the efficient and stable capture of the immunomagnetic bead cells.

[0019] In one embodiment, as shown in FIG. 2, in step S30, the step of pressing the liquid bag 60 flat includes the following steps S301 and S302. S301: Determine whether the closing mechanism 30 of the pressing device 1 can be closed. As shown in FIGS. 5 and 6, the pressing device 1 includes a closing mechanism 30 attached to the magnetic platform 10 and a pressing mechanism 40 connected to the closing mechanism 30. When the closing mechanism 30 is closed, the liquid bag 60 is located in the accommodation space between the closing mechanism 30 and the magnetic platform 10. The closing mechanism 30 is attached to the magnetic platform 10, and the closing mechanism 30 can close or open the above-mentioned accommodation space (when the liquid bag 60 arranged in the accommodation space bulges and the protruding height becomes high, the closing mechanism 30 may be pushed up by the liquid bag 60 and cannot be closed. If it is forcibly closed, the liquid bag 60 may burst or the immunomagnetic bead cells or anucleated immune cells therein may be damaged.). Also, when there is no interference by other external forces, the closing mechanism 30 can move downward by being pulled downward by the pressing mechanism 40. Thereby, the size of the accommodation space for arranging the liquid bag 60 between the magnetic platform 10 and the closing mechanism 30 can be adjusted.

[0020] S302: When the closing mechanism 30 can be closed, control the pressing mechanism 40 to move the closed closing mechanism 30 downward, and press the liquid bag 60 placed in the accommodation space between the magnetic platform 10 and the closing mechanism 30 flat. As can be understood, when the closing mechanism 30 can be closed, the pressing mechanism 40 drives the closed closing mechanism 30 to move downward, presses the liquid bag 60 uniformly and flatly, and can increase the contact surface formed by the close contact between the liquid bag 60 and the magnetic platform 10, so that the liquid in the liquid bag 60 uniformly passes through the contact surface and is distributed and contacted with the magnetic platform 10.

[0021] Furthermore, as shown in FIGS. 5 and 6, the pressing device 1 further includes a lift mechanism 20 connected to the closing mechanism 30. After step S301, that is, after determining whether the closing mechanism 30 of the pressing device 1 can be closed, the following steps are further included. When the closing mechanism 30 cannot be closed because it is pushed up by the swollen liquid bag 60, the lift mechanism 20 is controlled to lift the closing mechanism 30, thereby increasing the accommodation space between the magnetic platform 10 and the closing mechanism 30. In this embodiment, when the liquid bag 60 is disposed in the accommodation space, if the swelling height of the liquid in the liquid bag 60 is high and the closing mechanism 30 cannot be closed because it is pushed up by the liquid bag 60 disposed in the accommodation space, the lift mechanism 20 is controlled to be activated to lift the closing mechanism 30. In this way, the distance between the magnetic platform 10 and the closing mechanism 30 becomes larger, and the accommodation space increases due to the increase in this distance.

[0022] When the closing mechanism 30 rises to a preset closable height, the closing mechanism 30 may be controlled to be closed. That is, when the closing mechanism 30 rises to the preset height, the accommodation space between the closing mechanism 30 and the magnetic platform 10 is sufficient, the closing mechanism 30 is not pushed up by the liquid bag 60, and it can be normally closed.

[0023] The pressing mechanism 40 is controlled to move the closed closing mechanism 30 downward, thereby pressing the liquid bag 60 placed in the accommodation space flat. That is, after the closing mechanism 30 is closed, the pressing mechanism 40 drives the closed closing mechanism 30 to move downward, presses the liquid bag 60 uniformly and flatly, increases the contact surface formed by the close contact between the liquid bag 60 and the magnetic platform 10, and realizes that the liquid in the liquid bag 60 passes through the contact surface uniformly and contacts and distributes on the magnetic platform 10.

[0024] In one embodiment, as shown in FIGS. 5 and 6, a lift through hole 1011 is provided in the magnetic platform 10. The lift mechanism 20 includes a lifting assembly 201 provided at the top of the magnetic platform 10, a lift rod 202 whose tip penetrates through the lift through hole 1011 and is fixedly connected to the lifting assembly 201, and an upward drive assembly 203 provided below the magnetic platform 10 and connected to the end of the lift rod 202 away from the lifting assembly 201. The closing mechanism 30 includes a cover 301 rotatably connected to the lifting assembly 201. The shape of the cover 301 may be set according to the specific shape of the magnetic platform 10. Further, the closing mechanism 30 further includes a handle 3011 attached to the cover 301, and the user can easily pull or close the cover 301 by the handle 3011. The shape of the lifting assembly 201 may be set as needed. For example, the lifting assembly 201 may be a frame structure placed on the top of the magnetic platform 10 (or may be composed of a number of separately provided assemblies, but may also be driven by the lift rod 202 to move up and down), and may be driven by the upward drive assembly 203 to move up and down. As can be understood, the upward drive assembly 203 may include any one of a motor 2031, an air cylinder, etc. as a source of the driving force for lifting the lifting assembly 201. In one embodiment, as shown in FIGS. 5 and 6, the lifting assembly 201 includes a first lifting block 2011 provided at the first end of the magnetic platform 10, two second lifting blocks 2012 spaced apart and distributed at the second end of the magnetic platform 10 away from the first end, a limiting plate 2013 connected between the first lifting block 2011 and the second lifting blocks 2012, and a rotating shaft 2014 connected between the two second lifting blocks 2012. The cover 301 is rotatably connected to the rotating shaft 2014, and the second adsorption portion 303 shown in FIG. 5 is provided on the first lifting block 2011.Specifically, the limiting plate 2013 is composed of two plates provided on opposite sides of the magnetic platform 10 so as to be parallel. In this way, a limiting frame surrounded by the first lifting block 2011, the two second lifting blocks 2012, and the two limiting plates 2013 is obtained, which is used to prevent the liquid bag 60 from slipping out from the side of the accommodation space. The limiting frame (i.e., the main component of the lifting assembly 201) can move up and down in synchronization with the cover 301.

[0025] Furthermore, the step of increasing the accommodation space between the magnetic platform 10 and the closing mechanism 30 by controlling the lifting mechanism 20 to lift the closing mechanism 30 includes controlling and starting the lifting drive assembly 203, and driving the lift rod 202 by the lifting drive assembly 203 to lift the lifting assembly 201 and the cover 301 along the lift through hole 1011 while sliding, so as to increase the accommodation space for arranging the liquid bag 60 between the magnetic platform 10 and the cover 301. That is, the closing mechanism 30 is rotatably attached to the lifting assembly 201 and moves as the lifting assembly 201 moves up and down. On the other hand, the lifting drive assembly 203 can move the lifting assembly 201 upward along the lift through hole 1011 via the lift rod 202. In this way, the cover 301 is moved upward, and further, the accommodation space for arranging the liquid bag 60 between the magnetic platform 10 and the cover 301 is increased, facilitating the closing of the closing cover 301.

[0026] Furthermore, as shown in FIGS. 5 and 6, the closing mechanism 30 further includes a first adsorption portion 302 provided on the cover 301 and a second adsorption portion 303 provided at a position of the lifting assembly 201 facing the first adsorption portion 302. The cover 301 is closed with respect to the lifting assembly 201 through adsorption by the first adsorption portion 302 and the second adsorption portion 303. Here, at least one of the first adsorption portion 302 and the second adsorption portion 303 is a magnet. For example, both may be magnets, or one may be a magnet while the other is a metal having the property of being adsorbed by a magnet. The combination of the numbers, the specific shapes, and the sizes of the above-mentioned first adsorption portion 302 and second adsorption portion 303 may be set as required, but are not limited here. Furthermore, in the present application, the magnet may be an electromagnet. In one specific embodiment, the second adsorption portion 303 is an electromagnet, and the first adsorption portion 302 is an iron mass. In this case, when the electromagnet that is the second adsorption portion 303 is energized, the magnetism of the electromagnet can be released. At this time, since the electromagnet of the second adsorption portion 303 and the first adsorption portion 302 (iron mass) in the cover 301 have a magnetic adsorption function, they are in a separated state. Therefore, the cover 301 can be opened. On the other hand, when the energization of the second adsorption portion 303 stops, magnetism is generated in the electromagnet that is the second adsorption portion 303. At this time, if there is no external force interruption and the first adsorption portion 302 of the cover 301 is within the magnetic attraction range of the electromagnet, the first adsorption portion 302 of the cover 301 is adsorbed by the second adsorption portion 303, and the cover 301 is closed.

[0027] In one embodiment, as shown in FIGS. 5 and 6, the lifting drive assembly 203 includes a motor 2031, a drive wheel 2032, a driven wheel 2033, a timing belt 2034, a rotating shaft 2035, a cam 2036, a mounting bearing 2037, and a carrier 2038 having a mounting hole 2039. The carrier 2038 is attached to the bottom of the magnetic platform 10. The rotating shaft 2035 is mounted in the mounting hole 2039 through the mounting bearing 2037. The cam 2036 is fixedly attached to the rotating shaft 2035. The drive wheel 2032 is attached to the output shaft of the motor 2031. The timing belt 2034 is looped around the drive wheel 2032 and the driven wheel 2033. The driven wheel 2033 is fixedly attached to the rotating shaft 2035. The tip of the cam 2036 abuts against the lift rod 202. As shown in FIG. 5, the motor 2031 may be fixed to the magnetic platform 10 by a fixing block. The cam 2036, the carrier 2038, and the mounting bearing 2037 of the rotating shaft 2035 are all provided in two symmetrically. The lift rods 202 are also provided in two parallel to each other. The tips of the two lift rods 202 are all connected to the lifting assembly 201. On the other hand, the two cams 2036 are respectively abutted against the bottom ends of the two lift rods 202.

[0028] Furthermore, the step of driving the lift rod 202 by the ascending drive assembly 203 to raise the lifting assembly 201 and the cover 301 while sliding them along the lift through-hole 1011 includes the steps of controlling and starting the motor 2031, and driving the cam 2036 to rotate in sequence by the motor 2031 through the drive wheel 2032, the timing belt 2034, the driven wheel 2033, and the rotating shaft 2035, and raising the lifting assembly 201 and the cover 301 while sliding them along the lift through-hole 1011 through the lift rod 202 by the rotating cam 2036. That is, when the motor 2031 starts and rotates, it rotates the drive wheel 2032, and further rotates the driven wheel 2033 through the timing belt 2034. On the other hand, when the driven wheel 2033 rotates, the rotating shaft 2035 and the two cams 2036 attached to the rotating shaft 2035 rotate. At this time, the lift rod 202 in contact with the cam 2036 moves upward in conjunction with the rotation of the cam 2036, and further lifts the lifting assembly 201 and the cover 301 as a whole. At this time, as described in the above embodiment, when the second adsorption part 303 is an electromagnet, the energization of the electromagnet that is the second adsorption part 303 is stopped. In this way, in the ascending process of the lifting assembly 201, the cover 301 also gradually becomes flush with the lifting assembly 201 as it ascends. When the iron mass of the first adsorption part 302 is adsorbed by the electromagnet of the second adsorption part 303, it is determined that the cover 301 is closed.

[0029] Furthermore, as shown in FIG. 5, the ascending drive assembly 203 further includes a sensing block 2042 attached to the rotating shaft 2035 and a photoelectric sensor 2043 attached to the magnetic platform 10. When the photoelectric sensor 2043 detects that the rotating shaft 2035 drives the sensing block 2042 to rotate to a position facing the photoelectric sensor 2043, it is determined that the cover 301 has risen to the preset height. Here, the preset height refers to the highest height at which the lift rod 202 drives the lifting assembly 201 and the cover 301 to rise. This preset height may be set according to the specific size of the liquid bag 60. Specifically, it can be realized by changing the size and shape of the cam 2036 as needed.

[0030] In one embodiment, as shown in FIG. 5, the ascending drive assembly 203 further includes a bearing follower 2040 and a fixing plate 2041. The bearing follower 2040 is attached to the bottom end of the lift rod 202 by the fixing plate 2041, and the cam 2036 is in contact with the bearing follower 2040. That is, the bearing follower 2040 can transmit the lifting force generated by the rotation of the cam 2036 to the lift rod 202 well, making the ascent of the lift rod 202 more stable and controllable.

[0031] In one embodiment, as shown in FIGS. 5 and 6, the magnetic platform 10 is provided with a guiding through-hole 1012. The pressing mechanism 40 includes a guiding shaft 401, a spring 402, and a linear bearing 403 mounted in the guiding through-hole 1012. A stopper 4011 is provided at the bottom end of the guiding shaft 401. The tip of the guiding shaft 401 penetrates through the linear bearing 403 and is connected to the lifting assembly 201. The spring 402 is fitted onto the guiding shaft 401, and both ends of the spring 402 are respectively abutted against the linear bearing 403 and the stopper 4011. Further, in step S302, the step of controlling the pressing mechanism 40 to move the closed closing mechanism 30 downward includes controlling the upward driving assembly 203 to turn off, so that the biasing force of the spring 402 moves the guiding shaft 401 downward along the guiding through-hole 1012, and further includes the step of moving the lifting assembly 201 and the cover 301 downward. That is, in this embodiment, the upward driving assembly 203 having the cam 2036 can only drive the lift rod 202 to move upward, but does not have a downward pulling force to pull the lifting assembly 201 downward. Therefore, the above pressing mechanism 40 is provided, the spring 402 is in a compressed state, and after the motor 2031 of the upward driving assembly 203 is turned off (the motor 2031 is reset), the spring 402 resists its compressed state and applies a downward pulling force to the lifting assembly 201 through the guiding shaft 401 (the spring 402 drives the guiding shaft 401 to move downward along the guiding through-hole 1012), and further drives the lifting assembly 201 and the cover 301 to move downward.Understandably, if one guide shaft 401, one spring 402, and one linear bearing 403 are taken as a set of pressing assemblies, the above-mentioned pressing mechanism 40 may include a plurality of pressing assemblies provided on the magnetic platform 10, for example, four sets of pressing assemblies provided at the four corners of the rectangular magnetic platform 10. In this way, due to the downward pulling force applied by the four springs 402 of the four sets of pressing assemblies on the lifting assembly 201 at the same time, when the lifting assembly 201 moves downward, the cover 301 presses the liquid bag 60 towards the magnetic platform 10 until it becomes flat, and distributes the liquid therein evenly on the magnetic platform 10. In the present application, instead of a motor 2031 or the like, the spring 402 is pulled downward to press the liquid bag 60 flat. On the one hand, the spring 402 can press the liquid bag 60 evenly and flatly to make it adhere to the magnetic platform 10. On the other hand, since the biasing force of the spring 402 is uniform and small, it will not press and damage the cells in the liquid bag 60.

[0032] In one embodiment, as shown in FIGS. 5 and 6, the magnetic sorting and pressing device 1 further includes a protector 50 attached to the bottom end of the magnetic platform 10. An installation space is formed between the protector 50 and the magnetic platform 10, and the lifting drive assembly 203 is installed in the installation space. That is, in this embodiment, the protector 50 can house and protect each device (such as the lifting drive assembly 203, the pressing mechanism 40, or other devices) in the installation space. Preferably, the protector 50 is a sheet metal part with a stronger and more reliable structure.

[0033] It should be understood that the sequence numbers of the steps in the above embodiments do not mean the execution order, and the execution order of each process should be determined by its function and internal logic and does not limit the implementation process of the embodiments of the present application.

[0034] This application also provides a magnetic bead removal device including a pressing device 1 and a controller for executing the above magnetic bead removal method, wherein the controller is connected to the pressing device 1.

[0035] Specifically, the controller is used to execute the following steps: Placing a liquid bag having a curved flow path on a magnetic platform, wherein the plane where the curved flow path is located is above and parallel to the magnetic platform; Controlling a first flow rate control assembly to communicate the liquid bag with a sample container, and injecting a liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path; Pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved flow path and the magnetic platform, and adsorbing immunomagnetic bead cells in the curved flow path onto the contact surface by the magnetic platform; Performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container.

[0036] In one embodiment, the liquid bag includes a liquid inlet communicating with the sample container and a liquid outlet communicating with the collection container by the first flow rate control assembly, and the curved flow path communicates between the liquid inlet and the liquid outlet.

[0037] In one embodiment, the curved flow path includes a plurality of straight pipe segments arranged in parallel and a connecting pipe segment connected between two adjacent straight pipe segments, and the connecting pipe segment is a curved pipe or a straight pipe forming a preset inclination angle with the straight pipe segment, and / or The curved flow path includes a spiral pipe segment.

[0038] In one embodiment, the maximum width of the curved flow path is 50 mm or less, and / or The liquid bag further includes a buffer space, and the curved flow path communicates with the liquid outlet through the buffer space.

[0039] In one embodiment, the step of pressing the liquid bag flat is a step of determining whether the closing mechanism of the pressing device can be closed, the pressing device including a closing mechanism attached to the magnetic platform and a pressing mechanism connected to the closing mechanism, and when the closing mechanism is closed, the liquid bag is located in the accommodation space between the closing mechanism and the magnetic platform; when the closing mechanism can be closed, controlling the pressing mechanism to move the closed closing mechanism downward to press the liquid bag placed in the accommodation space between the magnetic platform and the closing mechanism flat, including.

[0040] In one embodiment, the pressing device further includes a lift mechanism connected to the closing mechanism, after the step of determining whether the closing mechanism of the pressing device can be closed, the controller further when the closing mechanism cannot be closed because it is pushed up by the inflated liquid bag, controlling the lift mechanism to raise the closing mechanism to increase the accommodation space between the magnetic platform and the closing mechanism; after the closing mechanism rises to a preset closable height, controlling the closing mechanism to close; and is used to execute the step of pressing the liquid bag placed in the accommodation space flat by controlling the pressing mechanism to move the closed closing mechanism downward.

[0041] In one embodiment, a lift through-hole is provided in the magnetic platform, and the lift mechanism includes a lifting assembly provided at the top of the magnetic platform, a lift rod whose tip penetrates the lift through-hole and is fixedly connected to the lifting assembly, and a lifting drive assembly provided below the magnetic platform and connected to an end of the lift rod away from the lifting assembly. The closing mechanism includes a cover rotatably connected to the lifting assembly. The step of increasing the accommodation space between the magnetic platform and the closing mechanism by controlling the lift mechanism to raise the closing mechanism is including the step of controlling and activating the lifting drive assembly, and driving the lift rod by the lifting drive assembly to raise the lifting assembly and the cover while sliding along the lift through-hole, so as to increase the accommodation space for arranging the liquid bag between the magnetic platform and the cover.

[0042] In one embodiment, the lifting drive assembly includes a motor, a drive wheel, a driven wheel, a timing belt, a rotating shaft, a cam, a mounting bearing, and a carrier having a mounting hole. The carrier is mounted at the bottom of the magnetic platform. The rotating shaft is mounted in the mounting hole by the mounting bearing. The cam is fixedly mounted on the rotating shaft. The drive wheel is mounted on the output shaft of the motor. The timing belt is wound around the drive wheel and the driven wheel. The driven wheel is fixedly mounted on the rotating shaft. The tip of the cam abuts against the lift rod. The step of driving the lift rod by the lifting drive assembly to raise the lifting assembly and the cover while sliding along the lift through-hole is Controlling and starting the motor, and driving the cam to rotate in the order of the drive wheel, the timing belt, the driven wheel, and the rotating shaft by the motor, and raising the lifting assembly and the cover along the lift through hole while sliding them through the lift rod by the rotating cam.

[0043] In one embodiment, the step of performing the collection operation to collect non-magnetic immune cells in the curved flow path into a collection container is Controlling and starting a second flow rate control assembly provided between the collection container and the liquid bag, communicating the liquid bag and the collection container, and transporting non-magnetic immune cells in the curved flow path to the collection container by the second flow rate control assembly.

[0044] For more detailed limitations on the pressing device 1 and the controller of the magnetic bead removal device, reference can be made to the limitations on the above magnetic bead removal method, and thus will not be elaborated here. Each module of the above controller may be implemented in whole or in part by software, hardware, and combinations thereof. Each of the above modules may be incorporated into the processor of a computer device in the form of hardware, or may be independent of this processor. Also, it may be stored in the memory of a computer device in the form of software, and called by the processor to execute operations corresponding to each of the above modules. As can be understood, the controller may be regarded as one or more computer devices. As shown in FIG. 8, the computer device includes a processor, a memory, a network interface, and a database connected via a system bus. The processor of the computer device provides computing and control capabilities. The memory of the computer device includes a readable storage medium and an internal memory. The readable storage medium stores an operating system, computer-readable instructions, and a database. The internal memory provides an environment for the operation of the operating system and computer-readable instructions in the readable storage medium. The database of the computer device is used to store data used in the magnetic bead removal method in the above embodiments. The network interface of the computer device is communicably connected to an external terminal via a network. When the computer-readable instructions are executed by the processor, the magnetic bead removal method is realized. The readable storage medium according to this embodiment includes a non-volatile readable storage medium and a volatile readable storage medium.

[0045] In one embodiment, one or more readable storage media storing computer-readable instructions are provided. The readable storage media according to this embodiment includes a non-volatile readable storage media and a volatile readable storage media. The computer-readable instructions are stored in the readable storage media and, when executed by one or more processors, cause the one or more processors to execute the above magnetic bead removal method.

[0046] Specifically, when the computer-readable instructions are executed by one or more processors, placing a liquid bag having a curved flow path on a magnetic platform, wherein a plane where the curved flow path is located is positioned above the magnetic platform and parallel to the magnetic platform; controlling a first flow rate control assembly to communicate the liquid bag with a sample container, thereby injecting a liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path; flattening the liquid bag to increase a contact surface between a bottom surface of the curved flow path and the magnetic platform, and adsorbing immunomagnetic bead cells in the curved flow path onto the contact surface by the magnetic platform; performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container, are caused to be executed by one or more processors.

[0047] In one embodiment, the liquid bag includes a liquid inlet communicating with the sample container and a liquid outlet communicating with the collection container by the first flow rate control assembly, and the curved flow path communicates between the liquid inlet and the liquid outlet.

[0048] In one embodiment, the curved flow path includes a plurality of straight pipe segments arranged in parallel and connection pipe segments connected between two adjacent straight pipe segments, and the connection pipe segments are curved pipes or straight pipes forming a preset inclination angle with the straight pipe segments, and / or The curved flow path includes a spiral pipe segment.

[0049] In one embodiment, the maximum width of the curved flow path is 50 mm or less, and / or The liquid bag further includes a buffer space, and the curved flow path communicates with the liquid outlet through the buffer space.

[0050] In one embodiment, the step of pressing the liquid bag flat is a step of determining whether the closing mechanism of the pressing device can be closed, the pressing device including a closing mechanism attached to the magnetic platform and a pressing mechanism connected to the closing mechanism, and when the closing mechanism is closed, the liquid bag is located in the accommodation space between the closing mechanism and the magnetic platform; when the closing mechanism can be closed, controlling the pressing mechanism to move the closed closing mechanism downward to press the liquid bag placed in the accommodation space between the magnetic platform and the closing mechanism flat.

[0051] In one embodiment, the pressing device further includes a lift mechanism connected to the closing mechanism. After the step of determining whether the closing mechanism of the pressing device can be closed, when the computer-readable instructions are executed by one or more processors, further when the closing mechanism cannot be closed because it is pushed up by the inflated liquid bag, controlling the lift mechanism to raise the closing mechanism to increase the accommodation space between the magnetic platform and the closing mechanism; after the closing mechanism rises to a preset closable height, controlling the closing mechanism to close. Causing one or more processors to execute the step of flattening the liquid bag placed in the accommodation space by controlling the pressing mechanism to move the closed closing mechanism downward.

[0052] In one embodiment, the magnetic platform is provided with a lift through hole, and the lift mechanism includes a lifting assembly provided at the top of the magnetic platform, a lift rod whose tip penetrates the lift through hole and is fixedly connected to the lifting assembly, and an upward drive assembly provided below the magnetic platform and connected to an end of the lift rod away from the lifting assembly. The closing mechanism includes a cover rotatably connected to the lifting assembly. The step of increasing the accommodation space between the magnetic platform and the closing mechanism by controlling the lift mechanism to raise the closing mechanism is Including the step of controlling and activating the upward drive assembly, driving the lift rod by the upward drive assembly, and raising the lifting assembly and the cover while sliding them along the lift through hole, so as to increase the accommodation space for arranging the liquid bag between the magnetic platform and the cover.

[0053] In one embodiment, the upward drive assembly includes a motor, a drive wheel, a driven wheel, a timing belt, a rotating shaft, a cam, a mounting bearing, and a carrier having a mounting hole. The carrier is mounted at the bottom of the magnetic platform. The rotating shaft is mounted in the mounting hole by the mounting bearing. The cam is fixedly mounted on the rotating shaft. The drive wheel is mounted on the output shaft of the motor. The timing belt is wound around the drive wheel and the driven wheel. The driven wheel is fixedly mounted on the rotating shaft. The tip of the cam abuts against the lift rod. The step of driving the lift rod by the upward drive assembly and raising the lifting assembly and the cover while sliding them along the lift through hole is Control the motor to start it, and rotate the cam in the order of the drive wheel, the timing belt, the driven wheel, and the rotating shaft by the motor, and raise the lifting assembly and the cover along the lift through-hole while sliding them via the lift rod by the rotating cam. This includes the step of

[0054] In one embodiment, the step of performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container is Control and start a second flow rate control assembly provided between the collection container and the liquid bag, communicate the liquid bag with the collection container, and transport non-magnetic immune cells in the curved flow path to the collection container by the second flow rate control assembly. This includes the step of

[0055] Regarding the specific limitations of the readable storage medium, since the limitations of the above magnetic bead removal method can be referred to, detailed description is not provided here.

[0056] Stored in a non-volatile readable memory medium or a volatile readable memory medium, when executed, by computer-readable instructions including the flow of the embodiments of the above methods, those skilled in the art can understand that the related hardware can be instructed to implement all or part of the flow in the methods of the above embodiments. References to memory, storage, databases, or other media used in each embodiment according to the present application may include non-volatile and / or volatile memory. Non-volatile memory may include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM), or flash memory. Volatile memory may include random access memory (RAM) or external cache memory. By way of example and not limitation, RAM is available in various forms such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), dual data rate SDRAM (DDR SDRAM), extended SDRAM (ESDRAM), synchronous link (Synchlink) DRAM (SLDRAM), Rambus direct RAM (RDRAM), direct memory bus dynamic RAM (RDRAM), and Rambus dynamic RAM (RDRAM).

[0057] For the convenience and brevity of description, those skilled in the art will only take the above division of each functional unit and module as an example for explanation. However, in actual applications, if necessary, the above functional division can be carried out by different functional units and modules, that is, it can be clearly understood that the internal structure of the device can be divided into different functional units and modules to complete all or part of the above functions.

[0058] The above embodiments are only used to explain the technical solution of the present application and do not limit it. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that they may modify the technical solutions described in each of the foregoing embodiments or equivalently replace some of their technical features. However, these modifications or replacements do not deviate from the spirit and scope of the technical solutions of the corresponding technical solutions from the spirit and scope of the technical solutions of each embodiment of the present application and should all be included in the protection scope of the present application.

Description of Reference Numerals

[0059] 1 Pressing device, 10 Magnetic platform, 101 Base, 1011 Lift through hole, 1012 Guide through hole, 102 Magnetic block, 20 Lift mechanism, 201 Lifting assembly, 2011 First lifting block, 2012 Second lifting block, 2013 Limiting plate, 2014 Rotating shaft, 202 Lift rod, 203 Upward driving assembly, 2031 Motor, 2032 Driving wheel, 2033 Driven wheel, 2034 Timing belt, 2035 Rotating shaft, 2036 Cam, 2037 Mounting bearing, 2038 Carrier, 2039 Mounting hole, 2040 Bearing follower, 2041 Fixed plate, 2042 Sensing block, 2043 Photoelectric sensor, 30 Closing mechanism, 301 Cover, 3011 Handle, 302 First adsorption part, 303 Second adsorption part, 40 Pressing mechanism, 401 Guide shaft, 402 Spring, 403 Linear bearing, 4011 Stopper, 50 Protector, 60 Liquid bag, 61 Curved flow path, 611 Straight pipe segment, 612 Connecting pipe segment, 62 Liquid inlet, 63 Liquid outlet

Claims

1. A method for removing magnetic beads, comprising: placing a liquid bag having a curved flow path on a magnetic platform, wherein a plane where the curved flow path is located is positioned above the magnetic platform and parallel to the magnetic platform; injecting a liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path by controlling a first flow rate control assembly to communicate the liquid bag with the sample container; flattening the liquid bag to increase a contact surface between a bottom surface of the curved flow path and the magnetic platform, and adsorbing immunomagnetic bead cells in the curved flow path onto the contact surface by the magnetic platform; performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container.

2. The liquid bag includes a liquid inlet communicating with the sample container and a liquid outlet communicating with the collection container by the first flow rate control assembly, and the curved flow path communicates between the liquid inlet and the liquid outlet. The method for removing magnetic beads according to Claim 1.

3. The curved flow path includes a plurality of straight pipe segments arranged in parallel and a connecting pipe segment connected between two adjacent straight pipe segments, and the connecting pipe segment is a curved pipe or a straight pipe forming a preset inclination angle with the straight pipe segment, and / or The curved flow path includes a spiral pipe segment. The method for removing magnetic beads according to Claim 2.

4. The maximum width of the curved flow path is 50 mm or less, and / or The liquid bag further includes a buffer space, and the curved flow path communicates with the liquid outlet through the buffer space. The method for removing magnetic beads according to Claim 3.

5. The step of flattening the liquid bag includes: determining whether a closing mechanism of a pressing device can be closed, the pressing device including a closing mechanism attached to the magnetic platform and a pressing mechanism connected to the closing mechanism, and when the closing mechanism is closed, the liquid bag is positioned in an accommodation space between the closing mechanism and the magnetic platform. When the closing mechanism can be closed, controlling the pressing mechanism to move the closed closing mechanism downward, and flattening the liquid bag placed in the accommodation space between the magnetic platform and the closing mechanism. The magnetic bead removal method according to claim 1 includes this step.

6. The pressing device further includes a lift mechanism connected to the closing mechanism. After the step of determining whether the closing mechanism of the pressing device can be closed. When the closing mechanism cannot be closed because it is pushed up by the inflated liquid bag, controlling the lift mechanism to raise the closing mechanism, thereby increasing the accommodation space between the magnetic platform and the closing mechanism. After the closing mechanism has risen to a preset closable height, controlling the closing mechanism to close it. Controlling the pressing mechanism to move the closed closing mechanism downward, thereby flattening the liquid bag placed in the accommodation space. The magnetic bead removal method according to claim 5 further includes this step.

7. The magnetic platform is provided with a lift through hole. The lift mechanism includes a lifting assembly provided at the top of the magnetic platform, a lift rod whose tip penetrates the lift through hole and is fixedly connected to the lifting assembly, and a lifting drive assembly provided below the magnetic platform and connected to the end of the lift rod away from the lifting assembly. The closing mechanism includes a cover rotatably connected to the lifting assembly. The step of increasing the accommodation space between the magnetic platform and the closing mechanism by controlling the lift mechanism to raise the closing mechanism. Controlling the lifting drive assembly to start, driving the lift rod by the lifting drive assembly, and raising the lifting assembly and the cover while sliding along the lift through hole, thereby increasing the accommodation space for placing the liquid bag between the magnetic platform and the cover. The magnetic bead removal method according to claim 6 includes this step.

8. The ascending drive assembly includes a motor, a driving wheel, a driven wheel, a timing belt, a rotating shaft, a cam, a mounting bearing, and a carrier having a mounting hole. The carrier is attached to the bottom of the magnetic platform. The rotating shaft is attached to the mounting hole by the mounting bearing. The cam is fixedly attached to the rotating shaft. The driving wheel is attached to the output shaft of the motor. The timing belt is wound around the driving wheel and the driven wheel. The driven wheel is fixedly attached to the rotating shaft. The tip of the cam abuts against the lift rod. The step of driving the lift rod by the ascending drive assembly to raise the lifting assembly and the cover while sliding them along the lift through-hole is The step of controlling and starting the motor, driving the cam to rotate in sequence by the motor through the driving wheel, the timing belt, the driven wheel, and the rotating shaft, and raising the lifting assembly and the cover along the lift through-hole while sliding them through the lift rod by the rotating cam, according to the magnetic bead removal method of claim 7.

9. The step of performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container is The step of controlling and starting a second flow rate control assembly provided between the collection container and the liquid bag, communicating the liquid bag with the collection container, and transporting non-magnetic immune cells in the curved flow path to the collection container by the second flow rate control assembly, according to the magnetic bead removal method of claim 1.

10. A magnetic bead removal device, comprising a pressing device and a controller configured to perform the magnetic bead removal method according to any one of claims 1 to 9, wherein the controller is connected to the pressing device.

11. A computer-readable storage medium, in which computer-readable instructions for realizing the magnetic bead removal method according to any one of claims 1 to 9 are stored when executed by a processor.

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