A magnetic bead separation device for isolating adeno-associated viral particles

Abstract

The utility model relates to the technical field of magnetic bead separation device, specifically a kind of magnetic bead separation device for separating adenovirus particles, comprising: device main body;Electromagnet, electromagnet is fixedly connected in device main body;Cover plate, cover plate is located in the upside of device main body by lifting assembly;Stirring mechanism, stirring mechanism is located in cover plate;Thread, thread is located in the lower part of the circumferential surface of device main body;And recovery bottle, recovery bottle is screw-connected on the surface of thread;By using the present device, the volume of device overall is small, easy to move, suitable for carrying, can be applied in clinical site, field research and emergency medical scene, and applicability is strong.

Description

Technical Field

[0001] This utility model relates to the technical field of magnetic bead separation devices, and in particular to a magnetic bead separation device for separating adeno-associated virus particles. Background Technology

[0002] Adeno-associated virus (AAV) magnetic bead separation technology is a highly efficient and specific purification method that utilizes surface-modified magnetic microspheres to selectively capture AAV particles. The magnetic beads typically have a diameter of 50 nm to 5 μm, with a core of superparamagnetic material (such as Fe3O4) and a surface coupled with AAV capsid-specific ligands (such as antibodies, affinity peptides, or receptors). The separation process consists of three stages: binding, washing, and elution. After incubation with the magnetic beads, AAV binds to them through affinity interactions. Under an applied magnetic field (0.1-1T), the magnetic bead-AAV complex rapidly migrates to the magnetic field region (usually designed with a circumferential outer wall or bottom magnetic field), and impurities are removed with the supernatant. Two to four washes with buffer (containing Tris-HCl, NaCl, and detergent) remove contaminants such as host proteins. Finally, high-purity AAV is eluted by adjusting the pH or ionic strength. This technology offers significant advantages: 1) High purity (host protein residue <5%); 2) High recovery rate (70-90%); 3) Fast processing speed (30 minutes to 2 hours, far faster than ultracentrifugation); 4) Scalability, adaptable to sample processing from microliters to larger volumes. Key parameters include magnetic bead size (1μm beads balance velocity and loading), magnetic field gradient (HGMS technology can reach 10³T / m), and buffer formulation (e.g., adding 0.01% Tween-20 to reduce non-specific adsorption). Automated systems (such as KingFisher™) further improve reproducibility and throughput, making it one of the preferred solutions for large-scale AAV production in gene therapy, especially meeting the stringent cGMP requirements for purity and safety.

[0003] In existing technologies, magnetic bead separation devices used to separate adeno-associated virus (AAV) particles are large in size, inconvenient to move, and difficult to carry. AAV separation requires going to a laboratory equipped with the separation device, which limits its application in clinical settings, field research, and emergency medical scenarios. Utility Model Content

[0004] The purpose of this invention is to provide a magnetic bead separation device for separating adeno-associated virus (AAV) particles in order to solve the above-mentioned problems. This invention improves upon the previous invention's large size, inconvenience in movement and portability, and the requirement to travel to a laboratory equipped with the separation device for AAV separation, which limited its application in clinical settings, field research, and emergency medical scenarios.

[0005] This utility model achieves the above-mentioned objective through the following technical solution: a magnetic bead separation device for separating adeno-associated virus particles, comprising:

[0006] Main body of the device;

[0007] An electromagnet, which is fixedly connected inside the main body of the device;

[0008] A cover plate, which is mounted on the upper side of the main body of the device via a lifting assembly;

[0009] A stirring mechanism is disposed inside the cover plate;

[0010] A thread, wherein the thread is located on the lower part of the circumferential surface of the device body; and

[0011] A recycling bottle, wherein the recycling bottle is threaded onto the surface of the thread.

[0012] Preferably, the lifting assembly includes a support plate, a top seat, and an electric push rod. There are two support plates, both of which are fixedly connected to the upper end of the main body of the device. The top seat is fixedly connected to the upper ends of the two support plates. The electric push rod is fixedly connected inside the top seat, and the extended end of the electric push rod movably passes through the lower end of the top seat and is fixedly connected to the upper end of the cover plate.

[0013] Preferably, the stirring mechanism includes a rotating groove, a rotating shaft, a mixing blade, a first gear, a second gear, a first motor, and a power module. The rotating groove is formed inside a cover plate, the rotating shaft is rotatably connected inside the cover plate, the mixing blade is fixedly connected to the circumferential surface of the rotating shaft, the first gear is fixedly connected to the circumferential surface of the rotating shaft, the first motor is fixedly connected inside the cover plate, the second gear is fixedly connected to the output end of the first motor and meshes with the first gear, and the power module is fixedly connected inside the cover plate and electrically connected to the first motor.

[0014] Preferably, a valve body is rotatably connected to the lower part of the main body of the device, and a second motor is fixedly connected to the lower part of the main body of the device, with the output end of the second motor fixed to one end of the valve body.

[0015] Preferably, the lower end of the top seat is fixedly connected to two limiting rods, both of which are fixedly connected to the upper end of the main body of the device, and the cover plate is slidably connected to the circumferential surface of the two limiting rods.

[0016] Preferably, the diameter of the main body of the device is 10cm.

[0017] Preferably, the upper end of the top seat is fixedly connected to two rotating seats, and the surfaces of the two rotating seats are rotatably connected to handles.

[0018] The beneficial effects of this utility model are:

[0019] 1. In this solution, the device is small in size, easy to move and carry, and can be used in clinical settings, field research and emergency medical scenarios, making it highly applicable.

[0020] 2. In this scheme, the power module in the stirring mechanism is a built-in power supply. The power module is used to provide power to the first motor. When the first motor is running, it drives the second gear connected to its output end to rotate. When the second gear rotates, it drives the first gear to rotate. The first gear drives the rotating shaft to rotate. The mixing blades fixed on the surface of the rotating shaft also rotate at the same time. When the mixing blades rotate, they stir the solution in the main body of the device. During the stirring process, the magnetic beads and the solution are fully mixed, which is conducive to the binding of the magnetic beads and AAV capsid protein. Attached Figure Description

[0021] Figure 1 This is a first-view perspective perspective view of the present invention;

[0022] Figure 2 This is a second-view perspective perspective view of the present invention;

[0023] Figure 3 This is a first sectional view of the present invention;

[0024] Figure 4 This is a second sectional view of the present invention.

[0025] In the diagram: 1. Main body of the device; 2. Electromagnet; 3. Cover plate; 4. Rotating groove; 5. Rotating shaft; 6. Mixing blade; 7. First gear; 8. Second gear; 9. First motor; 10. Power module; 11. Valve body; 12. Second motor; 13. Recycling bottle; 14. Thread; 15. Support plate; 16. Top seat; 17. Limiting rod; 18. Rotating seat; 19. Electric push rod; 20. Handle. Detailed Implementation

[0026] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0027] In practical implementation: such as Figures 1-4 As shown, a magnetic bead separation device for separating adeno-associated virus particles includes:

[0028] Device body 1;

[0029] Electromagnet 2 is fixedly connected inside the main body 1 of the device;

[0030] Cover plate 3 is mounted on the upper side of the main body 1 of the device via a lifting assembly;

[0031] A stirring mechanism is located inside the cover plate 3;

[0032] Thread 14, thread 14 is provided on the lower part of the circumferential surface of the device body 1; and

[0033] Recycling bottle 13 is threaded onto the surface of thread 14.

[0034] In this embodiment: the main body 1 of the device is used for the binding and separation of magnetic beads and adeno-associated virus (AAV). The surface of the magnetic beads is modified with specific ligands, which can selectively bind to AAV capsid proteins. During the binding process, the operation of the stirring mechanism ensures that the specific ligands on the surface of the magnetic beads fully bind to the AAV capsid proteins. After binding, the operation of the electromagnet 2 generates a magnetic field on the inner wall of the main body 1. The magnetic field causes the magnetic beads to adhere tightly to the inner circumference of the main body 1, controlling the operation of the second motor 12. The second motor 12 drives the valve body 11 to rotate, and the main body 1 is connected to the recovery bottle 13. The solution enters the recovery bottle 13 through the valve body 11. Since the magnetic beads are adsorbed on the inner circumference of the main body 1, the magnetic beads are separated from the solution containing AAV capsid proteins. The device is equipped with an external power supply, which is electrically connected to the second motor 12, the electromagnet 2, and the electric push rod 19 to provide power for the operation of the device. By using this device, the overall size of the device is small, easy to move, and suitable for carrying. It can be applied in clinical settings, field research, and emergency medical scenarios, and has strong applicability.

[0035] like Figures 1-4 As shown, the lifting assembly includes a support plate 15, a top seat 16, and an electric push rod 19. There are two support plates 15, both of which are fixedly connected to the upper end of the main body 1 of the device. The top seat 16 is fixedly connected to the upper end of the two support plates 15. The electric push rod 19 is fixedly connected inside the top seat 16. The extended end of the electric push rod 19 movably passes through the lower end of the top seat 16 and is fixedly connected to the upper end of the cover plate 3.

[0036] In this embodiment: the support plate 15 in the lifting assembly serves to support the top seat 16. An electric push rod 19 is installed inside the top seat 16. When the electric push rod 19 is running, it drives the cover plate 3 connected to its extended end to move and controls the position of the cover plate 3. When the magnetic beads bind to the AAV capsid protein inside the device body 1, the cover plate 3 slides into the device body 1 to seal the inside of the device body 1. Then, the stirring mechanism is controlled to run and stir the inside of the device body 1.

[0037] like Figures 1-4As shown, the stirring mechanism includes a rotating groove 4, a rotating shaft 5, a mixing blade 6, a first gear 7, a second gear 8, a first motor 9, and a power module 10. The rotating groove 4 is opened inside the cover plate 3. The rotating shaft 5 is rotatably connected inside the cover plate 3. The mixing blade 6 is fixedly connected to the circumferential surface of the rotating shaft 5. The first gear 7 is fixedly connected to the circumferential surface of the rotating shaft 5. The first motor 9 is fixedly connected inside the cover plate 3. The second gear 8 is fixedly connected to the output end of the first motor 9 and meshes with the first gear 7. The power module 10 is fixedly connected inside the cover plate 3 and is electrically connected to the first motor 9.

[0038] In this embodiment: the power module 10 in the stirring mechanism is a built-in power supply. The power module 10 is used to provide power to the first motor 9. When the first motor 9 is running, it drives the second gear 8 connected to its output end to rotate. When the second gear 8 rotates, it drives the first gear 7 to rotate. The first gear 7 drives the rotating shaft 5 to rotate. The mixing blade 6 fixed on the surface of the rotating shaft 5 also rotates at the same time. When the mixing blade 6 rotates, it stirs the solution in the main body 1 of the device. During the stirring process, the magnetic beads and the solution are fully mixed, which is beneficial to the binding of the magnetic beads and AAV capsid protein.

[0039] like Figures 1-4 As shown, a valve body 11 is rotatably connected to the lower part of the main body 1 of the device, and a second motor 12 is fixedly connected to the lower part of the main body 1 of the device. The output end of the second motor 12 is fixed to one end of the valve body 11.

[0040] In this embodiment: when the valve body 11 rotates, it is used to control the connection state between the main body 1 of the device and the recycling bottle 13. The second motor 12 is used to control the rotation of the valve body 11 and to control whether the main body 1 of the device and the recycling bottle 13 are connected.

[0041] like Figures 1-4 As shown, the lower end of the top seat 16 is fixedly connected to two limiting rods 17. Both limiting rods 17 are fixedly connected to the upper end of the main body 1 of the device, and the cover plate 3 is slidably connected to the circumferential surface of the two limiting rods 17.

[0042] In this embodiment, the limiting rod 17 is used to limit the cover plate 3, thereby improving the stability of the cover plate 3 moving up and down.

[0043] like Figures 1-4 As shown, the diameter of the main body 1 of the device is 10cm.

[0044] In this embodiment: the device body 1 of this diameter is small in size, suitable for carrying, and highly flexible.

[0045] like Figures 1-4 As shown, two rotating seats 18 are fixedly connected to the upper end of the top seat 16, and handles 20 are rotatably connected to the surfaces of the two rotating seats 18.

[0046] In this embodiment, the handle 20 facilitates hand movement of the device, making it easy to carry and improving its convenience.

[0047] It should be noted that the electromagnet 2, power module 10, first motor 9, electric push rod 19, second motor 12 and external power supply used in this device are all existing technologies. The specific models of electromagnet 2, power module 10, first motor 9, electric push rod 19, second motor 12 and external power supply used can be selected according to actual needs, and will not be elaborated on here.

[0048] In use, this invention first controls the lifting assembly to move the cover plate 3 upward, then adds a culture solution containing AAV capsid protein into the main body 1 of the device, and adds an appropriate amount of magnetic beads to the solution; controls the lifting assembly to move the cover plate 3 downward, closing the cover plate 3 with the main body 1 of the device; controls the operation of the stirring mechanism, the mixing blade 6 in the stirring mechanism rotates, agitating the solution, so that the magnetic beads and AAV capsid protein are fully combined; controls the operation of the electromagnet 2, the electromagnet 2 generates a magnetic field, causing the magnetic beads to adhere tightly to the inner circumference of the main body 1 of the device; controls the operation of the second motor 12, the second motor 12 drives the valve body 11 to rotate, and the solution enters the recovery bottle 13 through the valve body 11, completing the separation of the magnetic beads and the solution; by using this device, the overall size of the device is small, easy to move, suitable for carrying, and can be applied in clinical settings, field research, and emergency medical scenarios, with strong applicability.

[0049] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A magnetic bead separation device for separating adeno-associated virus particles, characterized in that, include: Device body (1); Electromagnet (2), which is fixedly connected inside the main body (1) of the device; Cover plate (3), which is located on the upper side of the main body (1) of the device via a lifting assembly; A stirring mechanism is provided inside the cover plate (3); Thread (14), said thread (14) being provided on the lower part of the circumferential surface of the device body (1); and A recycling bottle (13) is threaded onto the surface of a thread (14).

2. The magnetic bead separation device for separating adeno-associated virus particles according to claim 1, characterized in that: The lifting assembly includes a support plate (15), a top seat (16), and an electric push rod (19). There are two support plates (15), and both support plates (15) are fixedly connected to the upper end of the main body (1) of the device. The top seat (16) is fixedly connected to the upper end of the two support plates (15). The electric push rod (19) is fixedly connected inside the top seat (16). The extended end of the electric push rod (19) moves through the lower end of the top seat (16) and is fixedly connected to the upper end of the cover plate (3).

3. The magnetic bead separation device for separating adeno-associated virus particles according to claim 2, characterized in that: The stirring mechanism includes a rotating groove (4), a rotating shaft (5), a mixing blade (6), a first gear (7), a second gear (8), a first motor (9), and a power module (10). The rotating groove (4) is opened inside the cover plate (3). The rotating shaft (5) is rotatably connected inside the cover plate (3). The mixing blade (6) is fixedly connected to the circumferential surface of the rotating shaft (5). The first gear (7) is fixedly connected to the circumferential surface of the rotating shaft (5). The first motor (9) is fixedly connected inside the cover plate (3). The second gear (8) is fixedly connected to the output end of the first motor (9). The second gear (8) meshes with the first gear (7). The power module (10) is fixedly connected inside the cover plate (3). The power module (10) is electrically connected to the first motor (9).

4. A magnetic bead separation device for separating adeno-associated virus particles according to claim 3, characterized in that: A valve body (11) is rotatably connected to the lower part of the main body (1) of the device, and a second motor (12) is fixedly connected to the lower part of the main body (1). The output end of the second motor (12) is fixed to one end of the valve body (11).

5. A magnetic bead separation device for separating adeno-associated virus particles according to claim 4, characterized in that: The lower end of the top seat (16) is fixedly connected to two limiting rods (17), and both limiting rods (17) are fixedly connected to the upper end of the main body (1) of the device. The cover plate (3) is slidably connected to the circumferential surface of the two limiting rods (17).

6. A magnetic bead separation device for separating adeno-associated virus particles according to claim 5, characterized in that: The diameter of the main body (1) of the device is 10cm.

7. A magnetic bead separation device for separating adeno-associated virus particles according to claim 6, characterized in that: The upper end of the top seat (16) is fixedly connected to two rotating seats (18), and the surfaces of the two rotating seats (18) are rotatably connected to handles (20).

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