Magnetic bead sorting NK immune cell preparation and extraction device

Abstract

The utility model provides a magnetic bead sorting NK immune cell preparation and extraction device which comprises a base, a control shell is upwards arranged on the base, a magnetic field shell is arranged in the middle of the front side of the control shell, and a plurality of magnetic field grooves are inwards formed in the front side of the magnetic field shell. Each magnetic field groove comprises a first-level magnetic field groove located on the upper portion and a second-level magnetic field groove located on the lower portion, the section of each first-level magnetic field groove is larger than that of the corresponding second-level magnetic field groove, and a first-level magnetic field electromagnet is arranged on the periphery of each first-level magnetic field groove in the magnetic field shell. A secondary magnetic field electromagnet is arranged on the periphery of each secondary magnetic field groove in the magnetic field shell; the chromatographic column is divided into a sample section, a first-stage chromatographic section matched with the first-stage magnetic field groove and a second-stage chromatographic section matched with the second-stage magnetic field groove from top to bottom. The utility model provides a grading magnetic bead sorting structure, which can effectively avoid the blocking phenomenon, effectively improve the sorting efficiency of cells and improve the final yield.

Description

Technical Field

[0001] This invention belongs to the field of cell extraction technology and relates to a magnetic bead sorting device for preparing and extracting NK immune cells. Background Technology

[0002] In the biomedical field, the isolation and purification of immune cells are crucial for basic research and clinical treatment. Natural killer cells (NK cells), in particular, are an important component of the human immune system and play a key role in tumor immune surveillance and antiviral infection. NK cells can recognize and directly kill tumor cells or infected cells without prior sensitization, making them important targets in cancer immunotherapy.

[0003] Traditionally, methods for isolating NK cells include density gradient centrifugation and flow cytometry, but these methods are often complex, costly, and have limited efficiency. With the development of magnetic bead sorting technology, the technique of using specifically antibody-labeled magnetic beads to bind target cells, and then separating the target cells from a mixed cell suspension using a magnetic field, has gradually become one of the main methods for NK cell isolation. This technology not only improves the purity and recovery rate of cell isolation but also simplifies the operation process and reduces costs.

[0004] However, existing magnetic bead sorting devices still face certain challenges in practical applications. Especially when using chromatography columns for magnetic bead sorting, limitations in the design of existing chromatography columns, such as the relatively small diameter of the chromatography section, lead to a large accumulation of cells within the chromatography section during forward attraction, easily causing blockage. This blockage not only affects the continuity of the sorting process but also significantly reduces the sorting efficiency and final yield of NK cells. Utility Model Content

[0005] The purpose of this invention is to provide a magnetic bead sorting device for preparing and extracting NK immune cells. The device uses a graded magnetic bead sorting structure, which can effectively avoid clogging and improve the sorting efficiency and final yield of cells.

[0006] To solve the above-mentioned technical problems, this utility model provides a magnetic bead sorting NK immune cell preparation and extraction device, including a base, a control housing arranged upward at the rear of the base, a magnetic field housing arranged in the middle of the front side of the control housing, and multiple magnetic field grooves with openings at both the top and bottom of the front side of the magnetic field housing. Each magnetic field groove includes a primary magnetic field groove located at the top and a secondary magnetic field groove located at the bottom. The cross-section of each primary magnetic field groove is larger than the cross-section of the corresponding secondary magnetic field groove. A primary magnetic field electromagnet is arranged on the outer periphery of each primary magnetic field groove inside the magnetic field housing, and a secondary magnetic field electromagnet is arranged on the outer periphery of each secondary magnetic field groove inside the magnetic field housing.

[0007] A supplementary magnetic sheet is horizontally slidably connected to one side of each magnetic field slot on the front side of the magnetic field housing. Each supplementary magnetic sheet can cover the corresponding magnetic field slot. A strip-shaped magnetic drive shell is provided on the side of each supplementary magnetic sheet away from the corresponding magnetic field slot. A drive electromagnet that cooperates with the corresponding supplementary magnetic sheet is installed in each strip-shaped magnetic drive shell.

[0008] It also includes a chromatography column with openings at both the top and bottom. The chromatography column is divided from top to bottom into a sample section, a primary chromatography section, and a secondary chromatography section. The outer diameter of the sample section is larger than the outer diameter of the primary chromatography section. The outer diameter of the primary chromatography section matches the primary magnetic field cell, and the outer diameter of the secondary chromatography section matches the secondary magnetic field cell.

[0009] By adopting the above technical solution, blood samples are first collected, and peripheral blood mononuclear cells are separated by density gradient centrifugation and other methods. The separated PBMCs are resuspended in an appropriate buffer, and then a pre-prepared antibody-magnetic bead complex is added. After incubation, one or more washing steps are performed to remove unbound magnetic beads and antibodies.

[0010] The chromatography column is placed in the magnetic field tank. The primary, secondary, and drive electromagnets are activated. The drive electromagnet generates a magnetic field that repels the supplementary magnetic sheet, causing it to move outward and cover the magnetic field tank. This separates the NK cells bound to the magnetic beads from the mixed cell population and adds them to the sample section of the chromatography column. Under the action of the primary electromagnet and the supplementary magnetic sheet, the target NK cells of the magnetic beads gradually move towards the inner wall of the primary chromatography section without moving downward due to gravity, forming a sparse center and dense edge. This allows non-target cells and some of the target NK cells to continue downward into the secondary chromatography section. In the secondary chromatography section, the target NK cells of the magnetic beads also move towards the edge without moving downward due to gravity. Furthermore, the target NK cells of the magnetic beads are relatively sparse in the secondary chromatography section, making it easier for non-target cells to pass through, thus achieving rapid downward elution of non-target cells. Finally, the magnetic field is removed, and the NK cells are eluted from the column using an appropriate buffer or culture medium.

[0011] The present invention is further configured to include a test tube rack placed on the upper end of the base and located below the magnetic field housing, the test tube rack having a plurality of test tube insertion holes corresponding to the positions of the magnetic field slots.

[0012] The present invention is further configured such that the left and right sides of the base are provided with supporting edges that contact the lower end of the magnetic field shell, and the distance between the two supporting edges is equal to the length of the test tube rack.

[0013] The present invention is further configured such that a control button is provided at the upper end of the control housing at each magnetic field slot, and each control button is used to control the primary magnetic field electromagnet, the secondary magnetic field electromagnet and the drive electromagnet at the corresponding magnetic field slot.

[0014] The present invention is further configured such that a slide rail is provided on the upper and lower sides of each supplementary magnetic sheet on the front side of the magnetic field housing, and a slide bar is provided on the upper and lower sides of each supplementary magnetic sheet to be slidably connected to the corresponding slide rail.

[0015] The present invention is further configured such that each supplementary magnetic sheet has a push-pull magnetic strip arranged outward on the side of the corresponding strip magnetic drive shell.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] Firstly, this invention employs a two-stage magnetic bead sorting method. The chromatography column has a primary chromatography section and a secondary chromatography section. The outer diameter of the primary chromatography section is larger than that of the secondary chromatography section. During magnetic bead sorting, the target cells of the magnetic beads form a sparse-in-the-middle and dense-at-the-edge pattern in the primary chromatography section, allowing non-target cells and some target cells of the magnetic beads to continue downwards into the secondary chromatography section. Furthermore, in the secondary chromatography section, the target cells of the magnetic beads are relatively sparse, making it easier for non-target cells to pass through. This achieves rapid downward elution of non-target cells, effectively avoiding clogging and significantly improving cell sorting efficiency and final yield.

[0018] Secondly, by supplementing the controllable movement of the magnetic sheet, this utility model ensures that a uniform magnetic field exists around the chromatography column during magnetic bead sorting. This allows the target cells of the magnetic beads to approach the inner wall of the chromatography column evenly, rather than just clustering together on a portion of the inner wall, which helps to improve the sorting effect and avoid clogging. Attached Figure Description

[0019] Figure 1 This is a schematic diagram of the overall structure of this utility model;

[0020] Figure 2 It is a partial cross-sectional view used to show the internal structure of the magnetic field shell;

[0021] Figure 3 A schematic diagram of the overall structure of the chromatography column.

[0022] The components include: 1. Base; 11. Support side; 2. Control housing; 3. Magnetic field housing; 31. Primary magnetic field electromagnet; 32. Secondary magnetic field electromagnet; 33. Slide rail; 34. Strip magnetic drive housing; 35. Drive electromagnet; 4. Magnetic field slot; 41. Primary magnetic field slot; 42. Secondary magnetic field slot; 5. Supplementary magnetic sheet; 51. Slide bar; 52. Push-pull magnetic strip; 6. Control button; 7. Chromatography column; 71. Sample section; 72. Primary chromatography section; 73. Secondary chromatography section; 8. Test tube rack; 81. Test tube insertion hole. Detailed Implementation

[0023] The present invention provides a magnetic bead-sorted NK immune cell preparation and extraction device in further detail below with reference to the accompanying drawings and specific embodiments. The advantages and features of the present invention will become clearer from the following description. It should be noted that the drawings are all in a very simplified form and use non-precise proportions, and are only used to facilitate and clarify the illustration of the embodiments of the present invention. The same or similar reference numerals in the drawings represent the same or similar parts.

[0024] Example, refer to Figure 1-3 A magnetic bead sorting device for preparing and extracting NK immune cells includes a base 1. A control housing 2 is mounted on the rear of the base 1, and a circuit control module is installed inside the control housing 2. A magnetic field housing 3 is located in the middle of the front side of the control housing 2. Five magnetic field grooves 4 with openings at both the top and bottom are opened inward on the front side of the magnetic field housing 3. Each magnetic field groove 4 includes a primary magnetic field groove 41 located at the top and a secondary magnetic field groove 42 located at the bottom. The cross-section of each primary magnetic field groove 41 is larger than the cross-section of the corresponding secondary magnetic field groove 42. Three primary magnetic field electromagnets 31 are arranged in a circular pattern around the outer periphery of each primary magnetic field groove 41 inside the magnetic field housing 3. Three secondary magnetic field electromagnets 32 are arranged in a circular pattern around the outer periphery of each secondary magnetic field groove 42 inside the magnetic field housing 3.

[0025] A supplementary magnetic sheet 5 is horizontally slidably connected to one side of each magnetic field slot 4 on the front side of the magnetic field housing 3. A slide rail 33 is provided on the upper and lower sides of each supplementary magnetic sheet 5 on the front side of the magnetic field housing 3. A slide bar 51 is provided on the upper and lower sides of each supplementary magnetic sheet 5 and is slidably connected to the corresponding slide rail 33. Each supplementary magnetic sheet 5 can cover the corresponding magnetic field slot 4 and can form a four-sided enclosure with the primary magnetic field electromagnet 31 and the secondary magnetic field electromagnet 32. A strip-shaped magnetic drive shell 34 is provided on the side of each supplementary magnetic sheet 5 away from the corresponding magnetic field slot 4. A drive electromagnet 35 that cooperates with the corresponding supplementary magnetic sheet 5 is installed in each strip-shaped magnetic drive shell 34. A push-pull magnetic strip 52 is provided outward on the side of each supplementary magnetic sheet 5 close to the corresponding strip-shaped magnetic drive shell 34 to increase the area of ​​magnetic induction of the supplementary magnetic sheet 5. By changing the direction of the current passed into the drive electromagnet 35, the push-pull magnetic strip 52 is attracted or repelled.

[0026] A control button 6 is provided at the upper end of the control housing 2 at each magnetic field slot 4. Each control button 6 is used to control the primary magnetic field electromagnet 31, the secondary magnetic field electromagnet 32 ​​and the drive electromagnet 35 at the corresponding magnetic field slot 4 to achieve the sorting of one magnetic bead.

[0027] It also includes a chromatography column 7 with openings at both the top and bottom. The chromatography column 7 is divided from top to bottom into a sample section 71, a primary chromatography section 72, and a secondary chromatography section 73. The outer diameter of the sample section 71 is larger than the outer diameter of the primary chromatography section 72. The outer diameter of the primary chromatography section 72 matches the primary magnetic field cell 41, and the outer diameter of the secondary chromatography section 73 matches the secondary magnetic field cell 42.

[0028] It also includes a test tube rack 8 placed on the upper part of the base 1 and located below the magnetic field housing 3. The test tube rack 8 has two rows of test tube insertion holes 81 that correspond to the positions of the magnetic field slots 4. The left and right sides of the base 1 are provided with a support edge 11 that contacts the lower end of the magnetic field housing 3. The distance between the two support edges 11 is equal to the length of the test tube rack 8, which facilitates the quick alignment of the test tube insertion holes 81 with the magnetic field slots 4.

[0029] Working principle: First, blood samples are collected and peripheral blood mononuclear cells (PBMCs) are separated by density gradient centrifugation and other methods. The separated PBMCs are resuspended in an appropriate buffer solution, and then a pre-prepared antibody-magnetic bead complex is added. After incubation, one or more washing steps are performed to remove unbound magnetic beads and antibodies.

[0030] The chromatography column 7 is placed in the magnetic field tank 4. A test tube rack 8 is placed above the base 1, and test tubes are placed in the test tube rack 8 below the chromatography column 7. The primary magnetic field electromagnet 31, the secondary magnetic field electromagnet 32, and the driving electromagnet 35 are activated. The driving electromagnet 35 is energized to generate a magnetic direction that repels the supplementary magnetic sheet 5, pushing the supplementary magnetic sheet 5 outward to cover the magnetic field tank 4. This separates the NK cells bound to the magnetic beads from the mixed cell population and adds them to the sample section 71 of the chromatography column 7. Under the action of the primary magnetic field electromagnet 31 and the supplementary magnetic sheet 5, the target NK cells of the magnetic beads are located in the primary chromatography section 71. 2. Gradually move towards the inner wall of the column instead of moving downwards due to gravity, forming a sparse center and dense edges. This allows non-target cells and some target NK cells from the magnetic beads to continue moving downwards into the secondary chromatography section 73. Within the secondary chromatography section 73, the target NK cells of the magnetic beads also move towards the edges instead of moving downwards due to gravity. Furthermore, the target NK cells of the magnetic beads are relatively sparse in the secondary chromatography section 73, making it easier for non-target cells to pass through. This achieves rapid elution of non-target cells into the test tube. Finally, the magnetic field is removed, and the NK cells are eluted from the column using appropriate buffer or culture medium.

[0031] It should also be noted that all terms such as "set up" and similar descriptive words in this application (especially the specification) indicate that two structures have or exist a connection relationship. However, the specific means by which the two are connected are not limited in detail, and are usually conventional connection methods. That is, the means should be understood as prior art and do not need to be elaborated. For example, "m is set up with n" only indicates that structure m has structure n, and whether the two are connected by welding, riveting, adhesive, or integral molding is within the scope of protection of this application. Similarly, "x is rotatably set up with y" only indicates that y and x can rotate relative to each other, and whether the two are connected by a bearing, or whether y directly passes through x and is rotatably connected to x, or other feasible methods, are all within the scope of protection of this application.

[0032] The above description is only a description of the preferred embodiment of the present utility model and is not intended to limit the scope of the present utility model in any way. Any changes or modifications made by those skilled in the art based on the above disclosure shall fall within the protection scope of the claims.

Claims

1. A magnetic bead sorting NK immune cell preparation extraction device comprising a base (1), the back side of the base (1) is provided upward with a control shell (2), characterized in that, A magnetic field housing (3) is provided in the middle of the front side of the control housing (2). The front side of the magnetic field housing (3) has multiple magnetic field slots (4) with openings at both the top and bottom. Each magnetic field slot (4) includes a first-level magnetic field slot (41) at the top and a second-level magnetic field slot (42) at the bottom. The cross-section of each first-level magnetic field slot (41) is larger than the cross-section of the corresponding second-level magnetic field slot (42). A first-level magnetic field electromagnet (31) is provided on the outer periphery of each first-level magnetic field slot (41) inside the magnetic field housing (3). A second-level magnetic field electromagnet (32) is provided on the outer periphery of each second-level magnetic field slot (42) inside the magnetic field housing (3). The front side of the magnetic field housing (3) is horizontally slidably connected to one side of each magnetic field groove (4), and each supplementary magnetic sheet (5) can cover the corresponding magnetic field groove (4). The magnetic field housing (3) is provided with a strip magnetic drive shell (34) on the side of each supplementary magnetic sheet (5) away from the corresponding magnetic field groove (4). Each strip magnetic drive shell (34) is equipped with a drive electromagnet (35) that cooperates with the corresponding supplementary magnetic sheet (5). It also includes a chromatography column (7) with openings at both the top and bottom. The chromatography column (7) is divided from top to bottom into a sample section (71), a primary chromatography section (72), and a secondary chromatography section (73). The outer diameter of the sample section (71) is larger than the outer diameter of the primary chromatography section (72). The outer diameter of the primary chromatography section (72) matches the primary magnetic field groove (41), and the outer diameter of the secondary chromatography section (73) matches the secondary magnetic field groove (42).

2. The device for extracting NK immune cells prepared by magnetic bead sorting according to claim 1, characterized in that, It also includes a test tube rack (8) placed on the upper end of the base (1) and located below the magnetic field housing (3), the test tube rack (8) having a plurality of test tube insertion holes (81) corresponding to the position of the magnetic field groove (4).

3. The device for extracting NK immune cells prepared by magnetic bead sorting according to claim 2, characterized in that, The base (1) has support edges (11) on both the left and right sides that are in contact with the lower end of the magnetic field shell (3), and the distance between the two support edges (11) is equal to the length of the test tube rack (8).

4. The device for extracting NK immune cells prepared by magnetic bead sorting according to claim 1, characterized in that, The upper end of the control housing (2) is provided with a control button (6) at each magnetic field slot (4). Each control button (6) is used to control the primary magnetic field electromagnet (31), the secondary magnetic field electromagnet (32) and the drive electromagnet (35) at the corresponding magnetic field slot (4).

5. The device for extracting NK immune cells prepared by magnetic bead sorting according to claim 1, characterized in that, The front side of the magnetic field housing (3) is provided with slide rails (33) on both the upper and lower sides of each supplementary magnetic sheet (5), and each supplementary magnetic sheet (5) is provided with a slide bar (51) that is slidably connected to the corresponding slide rail (33) on both the upper and lower sides.

6. The device of claim 1, wherein the device is used for the preparation of NK immune cells extracted from magnetic beads. Each supplementary magnetic sheet (5) has a push-pull magnetic strip (52) on the side closest to the corresponding bar magnetic drive shell (34).

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