Magnetic bead continuous separation system and control method

Abstract

The invention relates to the technical field of nano magnetic bead preparation, and discloses a magnetic bead continuous separation system and a control method. The system comprises a magnetic bead separation device which is of a hollow pipe structure, the pipe wall of the magnetic bead separation device can be magnetized and demagnetized, one end of the magnetic bead separation device is provided with a first liquid inlet and a second liquid inlet, and the other end of the magnetic bead separation device is provided with a first liquid outlet and a second liquid outlet; the stock solution tank is connected to the first liquid inlet through a first flow channel; the waste liquid tank is connected to the first liquid outlet through a fifth flow channel, and a first magnetic bead sensor is arranged on the fifth flow channel; a liquid inlet of the product tank is connected to the second liquid outlet through a sixth flow channel, and a second magnetic bead sensor is arranged on the sixth flow channel; a liquid inlet of the stirring kettle is connected to the second liquid outlet through a seventh flow channel, and a liquid outlet of the stirring kettle is connected to the first liquid inlet through a second flow channel; a liquid outlet of the washing liquid tank is connected to the first liquid inlet through a third flow channel and is connected to the second liquid inlet through a fourth flow channel. The system has the advantage of meeting the requirements of continuous, automatic and infinite batch separation and purification of the magnetic beads.

Description

Technical Field

[0001] The present application relates to the technical field of nanomagnetic bead preparation, and in particular to a magnetic bead continuous separation system and control method. Background Art

[0002] Magnetic beads are a micron- or nanometer-scale particle structure with good magnetic response characteristics. The core is usually composed of a magnetic inorganic material (such as ferroferric oxide), the outer layer is coated with a polymer or siliceous material, and specific functional groups, such as carboxyl or amino groups, are introduced on the surface, so that they have the ability to specifically bind to target molecules. Due to their high flexibility in magnetic control operability and functional modification, magnetic beads are widely used in many biological and industrial fields such as nucleic acid extraction, protein purification, cell sorting, immunoassays, and magnetically controlled drug delivery. After synthesis, magnetic beads often have problems such as uneven magnetism, residual impurities, and inconsistent particle size distribution. If they are not purified and screened, their magnetic responsiveness and binding specificity will be directly affected, thereby reducing their separation efficiency and reusability in subsequent applications. Therefore, after the preparation of the magnetic beads is completed, they usually need to be separated and enriched to eliminate particles with weak magnetism, abnormal size, or high impurity content, so as to improve the overall purity and functional consistency.

[0003] At present, the separation and purification of magnetic beads mainly rely on manual operation or simple magnetic stand devices. Common methods include dividing the magnetic bead suspension into centrifuge tubes, manually achieving magnetic adsorption and discarding the liquid on a fixed magnetic stand, and then completing rough screening through multiple rounds of washing operations. Each step is usually completed in sequence in different centrifuge tubes, magnetic stands or other containers. Not only is the process cumbersome and dependent on manual operation, but magnetic bead loss, capture residue and cross-contamination are prone to occur during the transfer process. In addition, due to the different standards for manually judging the washing end point, the product purity and recovery rate often fluctuate, making it difficult to meet the needs of large-scale, high-consistency industrial production. Summary of the Invention

[0004] In order to meet the needs of continuous, automated, and unlimited batch separation and purification of target magnetic beads from non-target magnetic beads and impurities, and to achieve precise control of the entire process of capture, washing, and collection under dynamic regulation, the present application provides a magnetic bead continuous separation system and control method.

[0005] In a first aspect, the present application provides a magnetic bead continuous separation system, which adopts the following technical solution: A magnetic bead continuous separation system, comprising: A magnetic bead separation device having a hollow tube structure and capable of magnetizing and demagnetizing the tube wall. A first liquid inlet and a second liquid inlet are provided at one end of the magnetic bead separation device, and a first liquid discharge port and a second liquid discharge port are provided at the other end. The first liquid inlet is farther away from the tube wall region capable of magnetizing and demagnetizing than the second liquid inlet, and the first liquid discharge port is farther away from the tube wall region capable of magnetizing and demagnetizing than the second liquid discharge port. A raw liquid tank for storing the raw liquid to be processed; the raw liquid tank is connected to the first liquid inlet through a first flow channel; A waste liquid tank, the waste liquid tank is connected to the first liquid discharge port through a fifth flow channel, and a first magnetic bead sensor is provided on the fifth flow channel; a product tank, the liquid inlet of the product tank is connected to the second liquid discharge port through a sixth flow channel, and a second magnetic bead sensor is provided on the sixth flow channel; A stirring kettle, wherein the liquid inlet of the stirring kettle is connected to the second liquid outlet through a seventh flow channel, and the liquid outlet of the stirring kettle is connected to the first liquid inlet through a second flow channel; a washing liquid tank, wherein the liquid outlet of the washing liquid tank is connected to the first liquid inlet through a third flow channel and is connected to the second liquid inlet through a fourth flow channel; The first flow channel, the second flow channel, the third flow channel, the fourth flow channel, the fifth flow channel, the sixth flow channel and the seventh flow channel are all provided with valves, and the first flow channel, the second flow channel, the third flow channel and the fourth flow channel are provided with pump bodies.

[0006] Optionally, a first valve is provided on the first flow channel, a second valve is provided on the second flow channel, and a third valve is provided on the third flow channel. One end of the first valve away from the raw liquid tank, one end of the second valve away from the stirring kettle and one end of the third valve away from the washing liquid tank are connected to one end of the fourth valve through the first pump body, and the other end of the fourth valve is connected to the first liquid inlet; a second pump body and a fifth valve are provided on the fourth flow channel; a flow sensor and a sixth valve are provided on the fifth flow channel; an eighth valve is provided on the sixth flow channel, and a ninth valve is provided on the seventh flow channel. An end of the eighth valve away from the product tank and an end of the ninth valve away from the waste liquid tank are connected to the second discharge port through the seventh valve.

[0007] Optionally, a liquid level sensor is provided in the stirring tank, and the liquid level sensor is correspondingly set with a second preset upper limit height and a second preset lower limit height.

[0008] Optionally, the magnetic bead separation device is used to separate the raw liquid to be separated containing target magnetic beads, the target magnetic beads are particles that can be effectively captured by the external magnetic field, and the non-magnetic components in the non-magnetic waste liquid are impurity components that are not captured by the external magnetic field.

[0009] In a second aspect, the present application provides a control method for a continuous magnetic bead separation system, which adopts the following technical solution: A control method for a continuous magnetic bead separation system, used in a continuous magnetic bead separation system, comprises the following steps: Single batch stock solution feeding step: controlling the third valve to close and the first valve to open, and detecting whether the concentration of target magnetic beads detected by the first magnetic bead sensor reaches a first preset magnetic bead concentration; if not, continuing the detection; if yes, proceeding to the next step; wherein the target magnetic beads are particles that can be effectively captured by the magnetic field applied by the magnetic bead separation device; Single batch magnetic bead pipetting steps: control the first pump to stop, the fourth valve to close, and the sixth valve to close; interlock control the fifth valve, the seventh valve, and the ninth valve to open, and control the magnetic bead separation device to demagnetize; interlock control the second pump to start, and start the stirring tank; obtain the liquid level detection signal of the liquid level sensor in the stirring tank, and determine whether the liquid level of the stirring tank is higher than a second preset upper limit height. If not, continue to detect; if yes, proceed to the next step; Single batch magnetic bead stirring and washing steps: control the second pump to stop; chain control the fifth valve to close, the seventh valve to close, and the ninth valve to close; start the stirring tank and stir for a predetermined time; Waste liquid discharge step: control the first valve to close, the second valve to open, the fourth valve to open, and the sixth valve to open; control the magnetic bead separation device to magnetize and start the first pump body; obtain the liquid level detection signal of the liquid level sensor in the stirring tank, and judge whether the liquid level of the stirring tank is lower than the second preset lower limit height, if not, continue the detection, if yes, proceed to the next step; cleanliness detection step: stop the stirring tank, and draw a two-dimensional data graph about time based on the concentration of target magnetic beads detected by the first magnetic bead sensor in the waste liquid discharge step; detect whether the peak of the data graph is lower than the second preset magnetic bead concentration, if not, return to the single batch magnetic bead pipetting step, if yes, proceed to the next step; The steps for pipetting a single batch of finished magnetic bead liquid are as follows: the first pump is controlled to stop, the fourth valve and the sixth valve are controlled to close, the fifth valve, the seventh valve and the eighth valve are controlled to open in a chain manner, the magnetic bead separation device is controlled to be demagnetized, the second pump is controlled to start, and the concentration of the target magnetic beads detected by the second magnetic bead sensor is detected to be lower than the third preset magnetic bead concentration. If not, the detection is continued, and if so, the next step is entered. Batch cycle steps: control the fifth valve to close, the seventh valve to close, and the eighth valve to close, control the second pump to stop, obtain the liquid level detection signal of the liquid level sensor in the raw liquid tank, and determine whether the liquid level in the raw liquid tank is lower than the first preset lower limit height. If not, return to the single batch raw liquid feeding step.

[0010] Optionally, the following steps are also included: Pipeline self-cleaning steps: control the raw liquid tank to stop working, start the mixing kettle, control the fifth valve to open, the seventh valve to open, and the ninth valve to open, and start the second pump body; obtain the liquid level detection signal of the liquid level sensor in the mixing kettle and determine whether it is higher than the second preset upper limit. When it reaches the second preset upper limit, control the fourth valve to open, the sixth valve to open, and start the first pump body; based on the liquid level detection signal, determine whether the liquid level in the mixing kettle is lower than the second preset lower limit. If not, continue to detect. If so, complete the pipeline self-cleaning.

[0011] Optionally, the following steps are also included: The presetting step includes setting a first set value, a second set value, and a third set value; setting a first preset lower limit height, a second preset upper limit height, and a second preset lower limit height; and setting a stirring speed and a stirring time of the raw liquid tank and the stirring kettle.

[0012] Optionally, before the single batch stock solution feeding step, the following steps are further included: Magnetic bead separator pre-filling steps: control the second valve KV-2 to close, the third valve KV-3 to open, the fourth valve KV-4 to open, and the sixth valve KV-6 to open, control the magnetic bead separator M1 to magnetize and then start the first pump body P1; check whether the flow sensor detects the flow passing, if not, continue to test, if yes, go to the next step.

[0013] In summary, this application includes at least one of the following beneficial technical effects: 1. Realize continuous, automated, and unlimited batch separation operations of magnetic beads, breaking through the problems of process fragmentation, large manual operation errors, limited system throughput, and reliance on manual parameter adjustment in traditional magnetic bead separation technology; the device can automatically control the entire process from raw material input, magnetic separation, washing and rinsing to separation and recovery of target magnetic bead liquid, significantly improving the efficiency and stability of the magnetic bead separation process.

[0014] 2. Since the continuous automated magnetic bead continuous separation system adopts a multi-parameter adjustable structure and supports modular magnetic component configuration, users can flexibly adapt to magnetic beads of different particle sizes and different magnetic response characteristics without replacing the main equipment, which significantly improves the adaptability of the device and the expansion capability of the processing scale, and greatly reduces labor costs.

[0015] 3. By introducing liquid level control, online detection and valve interlock logic into the washing process, the system can automatically determine and precisely control the washing steps, reduce over-washing or under-washing, ensure the recovery rate and purity of the target object are in the optimal range, and significantly improve the batch consistency and repeatability of magnetic bead separation products. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1This is a pipeline connection diagram of a magnetic bead continuous separation system in a certain embodiment of the present application.

[0017] Figure 2 It is a structural schematic diagram of a magnetic bead separation device in a certain embodiment of the present application.

[0018] Figure 3 This is an overall schematic diagram of a magnetic field generating device in a certain embodiment of the present application. Figure 1 .

[0019] Figure 4 This is an overall schematic diagram of a magnetic field generating device in a certain embodiment of the present application. Figure 2 .

[0020] Figure 5 This is a flow chart of a control method for a magnetic bead continuous separation system in a certain embodiment of the present application. Figure 1 .

[0021] Figure 6 This is a flow chart of a control method for a magnetic bead continuous separation system in a certain embodiment of the present application. Figure 2 .

[0022] Figure 7 This is a flow chart of a control method for a magnetic bead continuous separation system in a certain embodiment of the present application. Figure 3 .

[0023] Reference numerals: 1. Magnetic bead separator; 11. First liquid inlet; 12. Second liquid inlet; 13. First liquid discharge port; 14. Second liquid discharge port; 101, first flow channel; 102, second flow channel; 103, third flow channel; 104, fourth flow channel; 105, fifth flow channel; 106, sixth flow channel; 107, seventh flow channel; 2. Magnetic field generating device; 21. Anti-stripping layer; 22. Embedded magnetic layer; 221. Support plate; 222. Magnetic tile; 223. Limiting groove; 23. Shielding layer; 3. Driving device; 31. Motor module; 32. Support rod; 33. Transmission rod; 4. Bracket; 5. Support column; 6. Sliding structure. DETAILED DESCRIPTION

[0024] Embodiments of the present application are described in detail below, examples of which are illustrated in the accompanying drawings.

[0025] Throughout this specification, reference to the terms "certain embodiments," "one embodiment," "some embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0026] Magnetic beads are a type of magnetic microsphere, whose core is usually composed of a magnetic inorganic material (such as ferroferric oxide), and the shell is coated with a polymer material or a siliceous material. By introducing functional groups such as carboxyl, amino, and streptavidin on their surface, magnetic beads can be given the ability to specifically bind to biological molecules, making them widely used in biomedical and industrial fields such as nucleic acid extraction, protein purification, cell sorting, immunoassays, and magnetically controlled drug delivery. The size of magnetic beads is generally between nanometers and micrometers, and they have good magnetic responsiveness. They can quickly aggregate under the action of an external magnetic field, achieving efficient capture and transfer of target objects.

[0027] However, after magnetic beads are synthesized, magnetic inhomogeneity often exists within the system, meaning that the magnetic response strength varies between different particles. Furthermore, some beads may harbor unremoved reaction byproducts, free monomers, or inorganic impurities, all of which can affect the separation efficiency and binding specificity of the beads in practical applications. Therefore, rigorous separation and screening procedures are required to eliminate particles with weak magnetism, high impurity content, or sizes that deviate from the specified range.

[0028] In this process, the magnetic beads with different magnetic strengths can be effectively screened with the help of an external magnetic field. Taking common laboratory operations as an example, researchers usually divide the magnetic bead suspension into centrifuge tubes, dilute them by manual pipetting, and place them on a fixed magnetic stand. The magnetic field is used to capture some particles with sufficient magnetic responsiveness on the tube wall, and the remaining non-magnetic or low-magnetic impurities are discarded with the supernatant. Subsequently, the retained magnetic beads are resuspended in liquid, and rough enrichment and purification are achieved through multiple "capture-discard liquid-add liquid-mix" operations. If a magnetic rod device is used, manual or robotic arm cooperation is required to immerse the magnetic rod in the magnetic bead suspension, and after capture, it is transferred to the cleaning solution for further washing.

[0029] However, traditional magnetic bead purification processes have significant limitations. First, the process is highly fragmented, with each step often requiring manual labor. This can lead to increased bead loss and reduced processing efficiency. Furthermore, the operator burden is high, and the batch processing volume is relatively small.

[0030] In practical applications, although the existing magnetic bead continuous separation system can cover the needs of some standard processes, its overall design generally lacks systematic consideration of processing flexibility and scalability, especially when dealing with magnetic beads of different magnetic response levels or changing single batch processing volumes, there are obvious limitations. For example, when it is necessary to process micro-magnetic particles or expand the batch size, the existing system can usually only respond by replacing it with a magnet with stronger magnetic properties or increasing the size of the equipment, resulting in a complicated equipment structure, increased system costs and reduced operational stability. In addition, since the magnetic attraction efficiency is closely related to the magnetic field gradient distribution, blindly increasing the magnetic intensity often cannot effectively solve the problem of poor selectivity of magnetic beads, but may instead cause excessive enrichment of non-target particles.

[0031] Furthermore, the lack of automation in the washing process is a key pain point that needs to be addressed urgently with existing technologies. Traditional magnetic bead separation equipment often relies on manual liquid addition, mixing, and capture processes during washing operations, lacking reliable termination criteria. This is especially true when removing non-specific binding components, which require repeated washing to ensure the complete removal of residual impurities. During this process, the operator must rely on experience to determine whether the washing is sufficient, which can easily lead to over-washing or under-washing due to misjudgment, thus affecting the purity of the magnetic beads.

[0032] In order to solve the problems of low separation efficiency, unstable washing process and insufficient automation in the existing magnetic bead separation process, the present invention provides a magnetic bead continuous separation system. The overall structure of the system is as follows: Figure 1 As shown, it includes a magnetic bead separation device M1, a raw liquid tank R1, a waste liquid tank T3, a product tank T2, a stirring tank R2 and a washing tank T1. The various components are interconnected through multiple flow channels and are equipped with valve components for controlling process switching.

[0033] The magnetic bead separation device M1 contains a hollow flow channel, and the wall area of ​​the flow channel can stably provide and eliminate a controllable magnetic field to achieve the selective capture and release of magnetic bead particles. The magnetic bead separation device M1 is provided with a first liquid inlet 11 and a second liquid inlet 12 at one end, and a first liquid discharge port 13 and a second liquid discharge port 14 at the other end, wherein the first liquid inlet 11 and the first liquid discharge port 13 are respectively located on the side away from the magnetic field action area, and the second liquid inlet 12 and the second liquid discharge port 14 are located on the side closer to the magnetic field action area relative to the first liquid inlet 11 and the first liquid discharge port 13. By selecting and combining different liquid inlets and liquid discharge ports through the first liquid discharge port 13, the second liquid discharge port 14, the second liquid inlet 12 and the second liquid discharge port 14, the liquid to be treated with magnetic beads can be circulated in the same magnetic bead separation device M1, so as to achieve different treatment purposes such as capture, separation, and washing at different stages. The stock liquid tank R1 is used to store the stock liquid to be separated containing target magnetic beads, and is connected to the first liquid inlet 11 of the magnetic bead separation device M1 through the first flow channel 101.

[0034] The waste liquid tank T3 is connected to the first liquid outlet 13 of the magnetic bead separation device M1 via a fifth flow channel 105. This flow channel is equipped with a first magnetic bead sensor AE1 for real-time detection of the residual magnetic bead concentration in the waste liquid. The product tank T2 is connected to the second liquid outlet 14 via a sixth flow channel 106. This flow channel is also equipped with a second magnetic bead sensor AE2 for monitoring the concentration of target magnetic beads in the finished product liquid.

[0035] The stirred tank R2 is the actuator for the resuspension process. Its liquid inlet is connected to the second liquid outlet 14 via the seventh flow channel 107 to receive the separated and purified target magnetic bead solution. The liquid outlet is connected back to the first liquid inlet 11 via the second flow channel 102, completing the closed-loop washing operation of the target magnetic beads. The washing tank T1 is connected to the first liquid inlet 11 via the third flow channel 103 and to the second liquid inlet 12 via the fourth flow channel 104, providing the system with the ability to inject washing liquid through different paths.

[0036] In order to achieve precise control of the fluid transmission process, independent valves are provided on the first to seventh flow channels, and pump bodies are provided on the first to fourth flow channels, which are used to switch and control the opening and closing states of the liquid passages and the liquid flow in each operation stage.

[0037] First, the magnetic bead separation device M1 set up in the system adopts an adjustable modular magnetic structure, which allows it to be flexibly configured according to the processing volume in different processing tasks. By adding or removing magnetic modules or adjusting the magnetic field distribution length, the processing capacity of each batch in the processing task can be expanded or shrunk to adapt to multiple scenarios from small-scale experiments to large-scale preparation.

[0038] Secondly, the degree of automation of the washing process has been greatly improved. The liquid level of the stirring tank R2 in the system is monitored in real time by a liquid level sensor, which serves as the criterion for starting and stopping the washing cycle. It realizes automatic closed-loop control of the entire process from liquid transfer, magnetic bead resuspension to washing liquid replacement, significantly reducing dependence on manual operation. In conjunction with the online detection unit deployed at the outlet of the washing device, such as conductivity sensor or fluorescent probe, the system can evaluate the impurity concentration in the washing liquid in real time, and adjust the three-way valve in conjunction to control the washing rounds and the volume of the washing liquid, thereby realizing quantitative management and standardized scheduling of the washing process. The washing parameters are controlled with high precision, and the coefficient of variation within the system can be controlled below 1%. Compared with the fluctuation range of more than 15% in traditional manual operation, it significantly enhances the consistency and process stability between batches, and meets the quality system requirements of GMP and GLP in the pharmaceutical production field.

[0039] Specifically, refer to Figure 2The magnetic bead separation device M1 includes a magnetic bead separator 1, a magnetic field generating device 2 and a driving device 3. The three together constitute a closed separation channel under a controllable magnetic field environment, which can realize the automatic capture, separation and washing of the target magnetic beads in the raw liquid to be separated. The target magnetic beads are particles that can be effectively captured by the external magnetic field, and the non-magnetic components in the non-magnetic waste liquid are impurity components that are not captured by the external magnetic field.

[0040] Specifically, the magnetic bead separator 1 is a hollow structure, which is used to pass the raw liquid to be processed carrying the target magnetic beads. The magnetic field generating device 2 is arranged on the periphery of the magnetic bead separator 1, which is used to form a stable strong magnetic field inside the magnetic bead separator 1 to capture the target magnetic beads to the tube wall. The driving device 3 is linked with the magnetic field generating device 2, and is used to adjust the distance between the magnetic field generating device 2 and the side wall of the magnetic bead separator 1 to change the magnetic field strength inside the magnetic bead separator 1. In different embodiments, the magnetic bead separator 1 can be fixed by a bracket 4 of different forms. The form of the bracket 4 can be determined according to the layout of the magnetic bead separator 1. When the magnetic bead separator 1 is horizontal or vertical, the bracket 4 should be adaptively adjusted, which is not specifically limited in this application.

[0041] Reference Figure 2 , a first liquid inlet 11 and a second liquid inlet 12 are provided at one end of the magnetic bead separator 1, and a first liquid discharge port 13 and a second liquid discharge port 14 are provided at the other end. Among them, the first liquid inlet 11 and the second liquid inlet 12 are used to introduce the raw liquid to be separated and the washing liquid containing the target magnetic beads, thereby realizing step-by-step alternating liquid input. The first liquid discharge port 13 is used to discharge impurities and suspended waste liquid contained in the raw liquid to be processed, while the second liquid discharge port 14 is used to output the magnetic bead liquid containing the target magnetic beads. With the help of the above-mentioned interface configuration, the entire process of raw liquid treatment, magnetic bead capture, and impurity flushing can be completed in a single cavity structure.

[0042] In various embodiments of the present application, the specific arrangement of the magnetic bead separator 1 can be adjusted based on the angle between its axis and the horizontal plane. The spatial configuration of the magnetic bead separator 1 may include, but is not limited to, vertical arrangement, horizontal arrangement, and diagonal arrangement. The vertical arrangement may also be referred to as a vertical arrangement, and the horizontal arrangement may also be referred to as a horizontal arrangement.

[0043] In an exemplary embodiment, the magnetic bead separator 1 is placed horizontally, and the magnetic field generating device 2 is still attached to the adaptation area on the outside of the tube body. Under this arrangement, the first liquid inlet 11 and the second liquid inlet 12 are both arranged at one end of the magnetic bead separator 1, and the first liquid discharge port 13 and the second liquid discharge port 14 are arranged at the other end of the magnetic bead separator 1. By controlling the horizontal flow of the liquid to be separated in the tube cavity, the capture of the target magnetic beads can be completed continuously. Furthermore, the second liquid inlet 12 is arranged on the side of the magnetic bead separator 1 close to the magnetic field generating device 2 relative to the first liquid inlet 11, and the second liquid discharge port 14 is arranged on the side of the end of the magnetic bead separator 1 close to the magnetic field generating device 2 relative to the first liquid discharge port 13.

[0044] In a preferred embodiment, the edge of the second liquid inlet 12 is tangential to the tube wall of the magnetic bead separator 1, allowing the injected liquid to flow along the axial direction of the magnetic bead separator 1. The second liquid discharge port 14 is also configured in a similar manner to be tangential to the tube wall of the magnetic bead separator 1, so as to facilitate the axial guidance and discharge of the target magnetic beads. The advantage of this structural arrangement is that during the washing liquid injection stage, an axial impact is quickly generated through the second liquid inlet 12, so that the deposited target magnetic beads can be efficiently flushed to the second liquid discharge port 14.

[0045] In a preferred embodiment of the present application, the magnetic field generating device 2 is arranged outside the magnetic bead separator 1, and its structure and magnet arrangement are designed to generate a magnetic field in the magnetic bead separator 1 that gradually weakens from the tube wall to the middle of the tube body.

[0046] Preferably, the magnetic bead separator 1 can be divided into a magnetic bead separation zone and a magnetic bead capture zone according to the magnetic field action characteristics and the fluid channel layout. Both zones are arranged along the axial direction of the magnetic bead separator 1. The two ends of the magnetic bead separation zone are connected to the first liquid inlet 11 and the first liquid discharge port 13, which mainly undertake the functions of injecting the original liquid and discharging the impurity waste liquid; the two ends of the magnetic bead capture zone are respectively connected to the second liquid inlet 12 and the second liquid discharge port 14, which are used for the injection of washing liquid and the output of magnetic bead liquid respectively. Specifically, the magnetic bead separation zone constitutes the main fluid channel, in which the target magnetic beads will be attracted by the magnetic force and move toward the magnetic bead capture zone; and the magnetic bead capture zone is close to the magnetic field action area, has a higher magnetic field strength, and can capture and fix the migrated magnetic beads.

[0047] It should be noted that the sizes of the first liquid inlet 11 and the first liquid discharge port 13 do not strictly correspond to the width of the magnetic bead separation zone, nor do the sizes of the second liquid inlet 12 and the second liquid discharge port 14 strictly correspond to the width of the magnetic bead capture zone. The magnetic bead separation zone and magnetic bead capture zone do not actually exist; rather, they are functional zoning within the magnetic bead separator 1 based on the behavior of the magnetic beads in the magnetic bead separation solution.

[0048] During operation, the target magnetic beads in the liquid to be separated maintain a certain spatial dispersion between the particles in the magnetic bead separation zone, and will not form agglomerations or sedimentation. However, under the action of the magnetic field, they gradually move toward the magnetic bead capture zone. As they enter the magnetic bead capture zone, the magnetic beads are quickly captured to the surface of the tube wall near the magnetic field generator and pressed against the tube wall, forming a densely arranged layer of magnetic beads. The magnetic bead particles are fixed to each other by the force of the magnetic field. During the operation of releasing the magnetic field, the magnetic field strength rapidly weakens, and the capture force between the particles suddenly decreases to a state of near-zero concentration. At this time, only a certain volume of washing liquid needs to be injected, and the impact force of the liquid can be used to quickly desorb the magnetic beads from the tube wall and bring them into the fluid.

[0049] In various embodiments, the magnetic field generating device 2 and the driving device 3 can be configured as a single group to partially enclose the magnetic bead separator 1; or they can be configured as multiple groups arranged around the magnetic bead separator 1 to increase the effective area of ​​the magnetic field. In specific implementations, the multiple groups of magnetic field generating devices 2 can be sequentially distributed around the circumference of the magnetic bead separator 1, or sequentially arranged along the axial direction of the magnetic bead separator 1, or distributed in an array on the surface of the magnetic bead separator 1, forming a composite magnetic field system with directional and gradient characteristics.

[0050] Specifically, in one embodiment, the magnetic field generating device 2 adopts an all-inclusive configuration. That is, by combining two or more sets of magnetic field generating devices 2 with the driving device 3, the magnetic field generating device 2 forms a closed surrounding structure along the outer wall of the magnetic bead separator 1, fully covering the entire circumference of the magnetic bead separator 1. This all-inclusive arrangement can generate a higher intensity and more uniform magnetic field within the magnetic bead separator 1, ensuring that target magnetic beads at any position in the lumen are effectively attracted.

[0051] Correspondingly, in another embodiment that is easy to assemble, the magnetic field generating device 2 can adopt a semi-enclosed configuration, that is, one or more groups of magnetic field generating devices 2 are only set on part of the outer wall area of ​​the magnetic bead separator 1, so that while forming an effective magnetic field capture area, part of the space is reserved for device observation or other functional module expansion.

[0052] In different embodiments, the magnetic field generating device 2 can use the principle of electromagnet to generate a magnetic field in the magnetic bead separation device 1, or can use the principle of permanent magnet to generate a magnetic field in the magnetic bead separation device 1, and can be designed according to specific needs. The magnetic field generating device 2 can provide different magnetic field strengths (0.5T-80T), which can be selected according to different magnetic beads and separation requirements. The following is an example of the magnetic field generating device 2 based on the structure of the permanent magnet, with reference to Figure 3In an embodiment of the present application, the magnetic field generating device 2 includes an anti-stripping layer 21, an embedded magnetic layer 22 and a shielding layer 23 arranged in sequence from the inside to the outside. The three-layer structure is used to establish a suitable magnetic field environment inside the magnetic bead separator 1 to achieve the capture and release of the target magnetic beads.

[0053] Among them, the first layer is an anti-stripping layer 21, that is, an inner layer bonding structure, which is used to prevent the magnetic tiles 222 in the embedded magnetic layer 22 from falling off, and to make the magnetic field generating device 2 in close contact with the outer wall of the magnetic bead separator 1. The anti-stripping layer 21 is usually made of stainless steel material, which has good mechanical strength and magnetic flux penetration. On the one hand, it can resist external stress impact and prevent the risk of displacement or falling off of the magnetic tiles 222. On the other hand, it will not significantly block or weaken the magnetic flux lines, ensuring that the magnetic field can effectively penetrate into the interior of the separation tube. In other embodiments, the anti-stripping layer 21 can also be made of non-magnetic titanium alloy, engineering plastic or aluminum alloy, but any material that has good mechanical strength, wear resistance and magnetic field penetration performance can be used. The geometric contour of the anti-stripping layer 21 is adapted to the shape of the outer wall surface of the magnetic bead separator 1 to achieve a close-fitting coating effect.

[0054] It should be noted that in some other embodiments, when the embedded magnetic layer 22 can stably fix the magnetic tile 222 alone, the anti-stripping layer 21 can be removed, that is, the magnetic field generating device 2 includes a two-layer structure of the embedded magnetic layer 22 and the shielding layer 23 arranged in sequence from the inside to the outside.

[0055] The second layer is the embedded magnetic layer 22, the core functional layer of the magnetic field generating device 2. It is embedded with multiple sets of magnetic tiles 222. Common materials for magnetic tiles 222 include neodymium iron boron, samarium cobalt, or other high-performance rare earth magnets. The embedded magnetic layer 22 is also made of stainless steel to balance structural support and traditional magnetic field characteristics. Accordingly, the structure of the embedded magnetic layer 22 can be divided into two types: semi-enclosed and fully enclosed, depending on the magnetic field distribution requirements. The fully enclosed structure uses a continuously surrounding magnetic module to cover the magnetic bead separation device, significantly increasing the capture area.

[0056] The third layer is the shielding layer 23, the outermost protective layer of the magnetic field generating device 2. It is used to shield against external magnetic field leakage. This shielding layer 23 restricts the spread of magnetic flux lines beyond the embedded magnetic layer 22, preventing the magnetic field from interfering with surrounding electronic components, signal lines, or operating equipment. Furthermore, the shielding layer 23 provides a certain degree of protection against external mechanical impact.

[0057] Reference Figure 4In a further embodiment of the present application, the embedded magnetic layer 22 includes a support plate 221 and a plurality of magnetic tiles 222. The support plate 221 is used to support and secure the magnetic tiles 222. The support plate 221 is integrally or separately connected to the shielding layer 23. A plurality of limiting grooves 223 are formed on the support plate 221 to limit the movement of the magnetic tiles 222. Each magnetic tile 222 is fixed by being embedded in a corresponding limiting groove 223, thereby ensuring that the magnetic tiles 222 do not deviate or become loose during operation of the device.

[0058] In an optional embodiment of the present application, referring to Figure 2 , the driving device 3 includes a motor module 31, a transmission rod 33 and a support rod 32. The bracket 4 is located on both sides of the motor module 31 and is internally provided with a transmission rod 33. The motor module 31 is output through the transmission rod 33 and is transmitted to the transmission rod 33 in the bracket 4 through the helical gear. The rotation of the transmission rod 33 drives the sliding structure 6 screwed thereto to move upward, thereby pushing the magnetic field generating device 2 to translate up and down. In different embodiments, the motor module can be a cylinder, an electric push rod or other different forms. In addition, the transmission form of the transmission rod 33 here can also be specifically embodied in different forms, such as equivalent replacement with a worm gear and worm transmission, or equivalent replacement with a gear and rack transmission, or equivalent replacement with a hydraulic transmission, a screw transmission, etc. The motor module 31 is installed on a pillar located in the middle of the bracket 4, and the support rod 32 is arranged at the top of the pillar and is slidably connected to the top of the pillar in the axial direction of the pillar, playing a directional role in the translation of the magnetic field generating device 2.

[0059] In summary, the magnetic bead separation device M1 adopts a fully enclosed structure, and there are no traditional rotating moving parts such as stirring shafts and blades inside the overall cavity, and the internal space shape design has no dead angles, which effectively avoids the problem of magnetic bead retention or accumulation caused by the stirring mechanism, and improves the separation efficiency and sample recovery rate. In the prior art, many magnetic bead processing equipment using an oblique intubation design frequently interfere with the magnetic bead deposition area due to the stirring components, resulting in the retention of magnetic beads in the separation cavity, which in turn causes a decrease in yield or separation failure. However, the present application significantly reduces the interference with the target magnetic beads by streamlining the inner cavity structure and eliminating the mechanical stirring device, while maintaining the uniform distribution of the magnetic beads and sufficient contact with the magnetic field.

[0060] In the continuous magnetic bead separation system described herein, the stock liquid tank R1 serves as a storage unit for the liquid to be processed. Its structure should meet the storage stability and liquid supply continuity requirements for large quantities of magnetic bead raw materials. The stock liquid tank R1 is connected to the first liquid inlet 11 of the magnetic bead separation device M1 via a first flow channel 101. This tank R1 is used to deliver pre-configured stock liquid containing magnetic beads to the magnetic bead separation device M1 as needed for subsequent capture and separation of target magnetic beads.

[0061] Specifically, the raw liquid tank R1 is a non-process liquid storage container, which adopts a double elliptical head structure as a whole. Compared with conventional flat-bottom or single elliptical structure tanks, the double elliptical structure has a larger total volume under the same diameter conditions, which is particularly suitable for the long-term storage needs of large quantities of ceramic bead separation liquid. In terms of internal structure, the raw liquid tank R1 is equipped with multiple sets of stirring paddle devices, which are driven by an external motor. It can be started and stopped in conjunction with the system through a "one-button start" signal to achieve periodic stirring operation of the magnetic bead raw liquid in the tank, preventing the magnetic beads from settling and aggregating after being left still for a long time, thereby ensuring that the raw liquid to be treated transported to the magnetic bead separation device M1 always maintains a state of uniform particle distribution.

[0062] In various embodiments, the stock liquid tank R1 can be a conventional single-walled structure or a double-jacketed structure, forming an adjustable temperature control system. A temperature-regulating medium is introduced into the jacket via a heating and cooling cycler, enabling real-time temperature control of the magnetic bead stock liquid within the tank. To further ensure accurate temperature control, a temperature measuring instrument fixedly connected to the tank wall is installed within the tank. Its measurement results form a closed-loop linkage with the external temperature control system, ensuring that the temperature within the tank is always maintained within the set range.

[0063] To facilitate system cleaning, a spray port is installed on the top of the R1 raw liquid tank. Its position, coordinated with the angle of the tank wall, covers the majority of the tank's surface area, effectively flushing any residual liquid or magnetic beads adhering to the tank wall. A level gauge port is also located in the middle of the tank, allowing it to connect to a level sensor for real-time monitoring of the raw liquid being processed. If the liquid level falls below a preset lower limit, the sensor outputs an alarm signal, which, in conjunction with the system's pump and valve logic, forms a chain trigger mechanism, automatically terminating subsequent infusion and processing.

[0064] In the continuous magnetic bead separation system described herein, waste liquid tank T3 serves as a receiving and temporary storage unit for the supernatant during the magnetic separation process. It is connected to the first liquid outlet 13 of the magnetic bead separation device M1 via the fifth flow channel 105 and is used to collect waste liquid that does not contain target magnetic beads and is discharged during the separation process. This waste liquid primarily includes impurities not captured by the magnetic field and non-target particles in the magnetic bead stock solution.

[0065] The T3 waste liquid tank can be designed as a vertical or horizontal double-elliptical head container, with the specific structure flexibly adjusted according to installation space and throughput requirements. The tank is equipped with a liquid level gauge interface, which can be connected to an external liquid level sensor for dynamic monitoring of the liquid level in the tank. When the waste liquid level reaches the set upper limit, the interlocking alarm system automatically outputs a control signal, triggering the associated pumps, valves, or control devices to stop operation, thereby avoiding the risk of overflow caused by overfilling the tank or back pressure in the downstream pipeline.

[0066] A first magnetic bead sensor AE1 is provided on the fifth flow channel 105 for real-time detection of the concentration of residual target magnetic beads in the waste liquid flowing through this path. This sensor is configured for online continuous monitoring and can be linked with the data acquisition and control module to enable process evaluation of separation efficiency and feedback determination of magnetic bead loss. In various embodiments, the first magnetic bead sensor AE1 can be a spectrophotometer, particle size analyzer, clarity meter, turbidity meter, or other device. Although these instruments utilize different detection principles, the system processes their measurement data to indirectly measure the target magnetic bead concentration.

[0067] In the magnetic bead continuous separation system described in the present application, the product tank T2 is used to receive and store the target magnetic bead liquid product obtained after the separation and washing process, and its liquid inlet is connected to the second liquid discharge port 14 of the magnetic bead separation device M1 through the sixth flow channel 106, so that the target magnetic bead liquid can finally enter the storage link.

[0068] In different embodiments, the product tank T2 can be a common single-wall structure or a double-jacketed structure. The single-wall structure has the advantage of low cost, while the inner cavity of the double-jacket can be connected to a heating and cooling circulation system for precise control of the internal temperature of the tank. By setting a temperature sensor in the tank and implementing closed-loop control with the circulation device, the magnetic bead liquid can always be kept at a suitable constant temperature. To further improve the cleaning ability of the equipment, a spray cleaning interface is provided on the top of the tank, which can be used to implement internal flushing operations after the system operation is completed, effectively removing any magnetic bead liquid that may remain on the tank wall and avoiding cross-contamination of batches.

[0069] At the same time, a liquid level gauge interface is reserved on the outer wall of the product tank T2 for connecting a liquid level monitoring sensor to achieve real-time detection of changes in the volume of the tank. When the magnetic bead liquid filling reaches the preset upper limit liquid level, the system sends an alarm signal through the interlocking control logic and synchronously controls the front-stage fluid input path to stop running. A second magnetic bead sensor AE2 is also provided on the sixth flow channel 106 to monitor the concentration of the magnetic bead liquid entering the product tank T2. The measured real-time data can be used to determine the residual degree of the target magnetic beads in the magnetic bead separation device M1. In different embodiments, the second magnetic bead sensor AE2 can be a spectrophotometer, a particle size analyzer, a clarity detector, a turbidity detector, and other devices. The system processes its measurement data to obtain an indirect measurement of the magnetic bead concentration.

[0070] In the magnetic bead continuous separation system described in the present application, the stirring tank R2 serves as an intermediate container for the magnetic bead washing process, and its liquid inlet is connected to the second liquid discharge port 14 of the magnetic bead separation device M1 through the seventh flow channel 107, for receiving the magnetic bead liquid after preliminary purification; its liquid outlet is refluxed through the second flow channel 102 and connected to the first liquid inlet 11 of the magnetic bead separation device M1 to realize the cyclic washing operation of the magnetic beads.

[0071] The R2 stirred tank body is designed as a vertical structure with an upper elliptical head and a lower conical head. The design angle of the lower conical head is greater than 60°, that is, the repose angle is greater than the natural inclination angle formed by the accumulation of magnetic beads in a static state, which can effectively prevent the magnetic beads from accumulating and settling at the bottom of the tank, thereby reducing the amount of residual liquid and magnetic bead loss during the washing process.

[0072] The stirring kettle R2 is internally equipped with multiple sets of stirring paddles driven by a motor. The stirring module can be automatically operated by a "one-button start" method to evenly disperse the magnetic beads. In different embodiments, the stirring kettle R2 can also be a common single-layer wall structure or a double-layer jacket structure. The jacket is connected to the heat exchange medium output by the heating and refrigeration cycle machine, and a temperature sensor is provided to be inserted into the tank body to achieve real-time temperature monitoring and precise constant temperature control during the stirring process. A spray port is provided on the top of the stirring kettle R2 to facilitate the cleaning of the magnetic bead liquid hanging on the wall of the tank after batch changes or long-term shutdown.

[0073] Furthermore, to monitor the tank's liquid level in real time, the top of the stirred tank R2 is equipped with a level gauge interface and interlocking alarm mechanisms for both full and empty states. When the liquid level exceeds the specified limit or the tank is empty, the control system responds immediately, intervening through the interlocking mechanism to initiate start / stop commands for the upstream inlet or downstream outlet processes.

[0074] The washing tank T1 is used to store and supply the washing liquid required for the magnetic bead separation process. Its liquid outlet is connected to the first liquid inlet 11 of the magnetic bead separation device M1 through the third flow channel 103 and to the second liquid inlet 12 of the magnetic bead separation device M1 through the fourth flow channel 104. Through this connection method, the washing tank T1 can provide the initial washing liquid and the washing liquid for desorbing the magnetic beads to the magnetic bead separation device M1, respectively, to achieve continuous or multiple rounds of magnetic bead washing operations in the system. The washing liquid can be selected from different options depending on the raw material of the magnetic beads, such as deionized water, ethanol, saline solution, high molecular polymer, etc.

[0075] Washing tank T1 is also preferably a double-elliptical head container, which can be flexibly configured in a vertical or horizontal configuration according to the process layout. Two independent discharge ports are set at the bottom of the tank. One discharge port is connected to the centrifugal pump interface for efficiently extracting washing liquid before washing the magnetic beads; the other discharge port is used to provide a pre-liquid filling function for the magnetic bead separator, ensuring that the magnetic bead separator is pre-filled with liquid before the raw liquid to be processed is introduced, thereby optimizing the distribution state and desorption path of the magnetic beads under the action of the magnetic field.

[0076] In this embodiment, the system is provided with a plurality of valves and pump bodies, which are respectively arranged on a plurality of flow channels to control the flow direction and on-off relationship of different fluids. Specifically, the first flow channel 101 is used to transport the raw liquid to be separated in the raw liquid tank R1 to the magnetic bead separation device M1, and a first valve KV-1 is provided thereon, which is located near one end of the magnetic bead separation device M1 to control the injection process of the raw liquid. The end of the first flow channel 101 away from the raw liquid tank R1 is connected to the fourth valve KV-4 through the first pump body P1, and the other end of the fourth valve KV-4 is connected to the first liquid inlet 11 of the magnetic bead separation device M1, thereby forming a controlled delivery path for the raw liquid to be processed.

[0077] The second flow channel 102 is used to circulate the magnetic bead washing liquid in the stirring tank R2 back to the magnetic bead separation device M1. A second valve KV-2 is provided on it. The end of the second valve KV-2 away from the stirring tank R2 is connected to the inlet end of the above-mentioned fourth valve KV-4 through the first pump body P1.

[0078] The third flow channel 103 is used to transport washing liquid from the washing tank T1 to the first liquid inlet 11 of the magnetic bead separation device M1. A third valve KV-3 is provided on the third flow channel 103. The end of the third valve KV-3 away from the washing tank T1 is also connected to the fourth valve KV-4 through the first pump body P1, thereby merging with the first flow channel 101 and the second flow channel 102 near the magnetic bead separation device M1. The raw liquid to be processed and the washing liquid can be switched and input here.

[0079] The fourth flow channel 104 is used to deliver the liquid in the washing tank T1 to the second liquid inlet 12 of the magnetic bead separation device M1. A second pump body P2 and a fifth valve KV-5 are provided on the fourth flow channel 104. The second pump body P2 and the fifth valve KV-5 cooperate to control the injection amount of the washing liquid and provide driving force.

[0080] The fifth flow channel 105 is used to discharge the waste liquid from the first liquid discharge port 13 of the magnetic bead separation device M1 and guide it to the waste liquid tank T3, on which the first magnetic bead sensor AE1 and the sixth valve KV-6 are provided. The sixth valve KV-6 is used to perform fluid cut-off control before and after the waste liquid is discharged, and to determine the effective capture rate of the target magnetic beads in conjunction with the sensor signal to prevent the target magnetic beads from being discharged with the waste liquid.

[0081] The sixth flow channel 106 is used to connect the second liquid discharge port 14 of the magnetic bead separation device M1 to the product tank T2, on which the second magnetic bead sensor AE2 and the eighth valve KV-8 are provided. The eighth valve KV-8 is arranged on the side close to the magnetic bead separation device M1 to ensure that only magnetic bead liquid that meets the concentration standard is introduced into the product tank T2.

[0082] The seventh flow channel 107 is used to connect the second liquid discharge port 14 of the magnetic bead separation device M1 to the stirring tank R2, and a ninth valve KV-9 is provided on it. The end of the ninth valve KV-9 away from the stirring tank R2 and the end of the eighth valve KV-8 away from the product tank T2 are jointly converged into the seventh valve KV-7. The seventh valve KV-7 is connected to the second liquid discharge port 14 of the magnetic bead separation device M1. The three constitute the diversion control node of the product and the circulation flow path.

[0083] The pump bodies provided in each of the above-mentioned flow channels include a first pump body P1 (e.g., a peristaltic pump), which is deployed on the first flow channel 101, the second flow channel 102, and the third flow channel 103. The second pump body P2 (e.g., a centrifugal pump) is deployed on the fourth flow channel 104 to provide the flow driving capacity required at different stages. It should be noted that the flow channels can be independently provided in different embodiments or adaptively combined according to process requirements. For example, in an area near the magnetic bead separation device M1, the first flow channel 101, the second flow channel 102, and the third flow channel 103 can share a combined pipeline to simplify the pipeline structure, reduce the stagnant liquid volume, and improve the overall integration.

[0084] In the continuous automated magnetic bead continuous separation system described in the present application, the first pump body P1 is linked to the fourth valve KV-4 and the sixth valve KV-6. When the operation stops, the magnetic bead liquid can be discharged only from the second liquid discharge port 14 of the magnetic bead separation device M1 by closing the valve to avoid backflow. The second pump body P2 is used to provide a large flow liquid driving force when flushing the pipeline. The second pump body P2 is preferably a centrifugal pump. The centrifugal pump has good flow control performance and can accurately set the liquid output rate to match the flow resistance characteristics under different pipeline structures. In order to achieve automatic control, the second pump body P2 and the first pump body P1 are interlocked with each other. When the centrifugal pump is running, the first pump body P1 is in a stopped state to avoid the formation of interference flow or pressure fluctuations in the pipeline system. Furthermore, the second pump body P2 is linked to the fifth valve KV-5 and the seventh valve KV-7. When the second pump body P2 is stopped, only the waste liquid (supernatant) is allowed to flow out of the first drain port 13. When the second pump body P2 is started, the magnetic bead liquid is flushed out of the second drain port 14, achieving directional transfer of the magnetic beads. Furthermore, the start and stop operations of the second pump body P2 are logically linked to the magnetization / magnetization behavior of the magnetic field generator. When the magnetic bead release operation (magnetization) is executed, the second pump body P2 is activated; when the magnetic separation process (magnetization) is resumed, the second pump body P2 stops, and the system returns to the magnetic bead circulation washing process driven by the first pump body P1.

[0085] The system also includes multiple detection instruments, such as the first magnetic bead sensor AE1 and the second magnetic bead sensor AE2. These instruments include spectrophotometers, particle size analyzers, clarity meters, and turbidity meters, respectively, for monitoring parameters such as target concentration in the magnetic bead solution, magnetic bead particle size, solution clarity, and wash solution turbidity. These instruments then calculate the magnetic bead concentration based on the test results and automatically adjust the operating conditions of the pumps and valves. Furthermore, the first pump P1 can be replaced with a metering pump in various implementations or operated in conjunction with an independent flow control circuit to accommodate different processing requirements. The operating parameters of all pumps in the system can be set according to different separation conditions, including the feed rate of the magnetic bead solution and the flushing flow rate of the wash solution.

[0086] The present application also discloses a method for controlling a continuous magnetic bead separation system. Figure 5-Figure 7 , including the following steps: Device reset steps: respond to the reset operation instruction and control each component to enter the initial state; including controlling the first valve KV-1 to close, the second valve KV-2 to close, the third valve KV-3 to close, the fourth valve KV-4 to close, the fifth valve KV-5 to close, the sixth valve KV-6 to close, the seventh valve KV-7 to close, the eighth valve KV-8 to close, and the ninth valve KV-9 to close in sequence; controlling the first pump body P1 to stop and the second pump body P2 to stop; controlling the raw liquid tank R1 to stop and the stirring kettle R2 to stop; controlling the magnetic bead separation device M1 to perform demagnetization (demagnetization) operation.

[0087] Pre-setting steps: setting the first preset magnetic bead concentration, the second preset magnetic bead concentration, and the third preset magnetic bead concentration; setting the first preset lower limit height, the second preset upper limit height, and the second preset lower limit height; setting the stirring speed and stirring time of the stock liquid tank R1 and the stirring kettle R2.

[0088] Pre-filling steps of the magnetic bead separation device: control the second valve KV-2 to close, the third valve KV-3 to open, the fourth valve KV-4 to open, and the sixth valve KV-6 to open, control the magnetic bead separation device M1 to magnetize and then start the first pump body P1; check whether the flow sensor detects the flow passing, if not, continue to test, if yes, go to the next step.

[0089] Single batch stock solution feeding steps: control the third valve KV-3 to close, the first valve KV-1 to open, and detect whether the concentration of target magnetic beads detected by the first magnetic bead sensor AE1 reaches the first preset magnetic bead concentration. If not, continue the detection; if so, proceed to the next step; wherein, the target magnetic beads are particles that can be effectively captured by the magnetic field applied by the magnetic bead separation device M1.

[0090] Single batch magnetic bead pipetting steps: control the first pump body P1 to stop, the fourth valve KV-4 to close, and the sixth valve KV-6 to close; interlock control the fifth valve KV-5, the seventh valve KV-7, and the ninth valve KV-9 to open, and control the magnetic bead separation device M1 to demagnetize; interlock control the second pump body P2 to start, and the stirring tank R2 to start; obtain the liquid level detection signal of the liquid level sensor in the stirring tank R2, and determine whether the liquid level of the stirring tank R2 is higher than the second preset upper limit height. If not, continue to detect, and if so, proceed to the next step.

[0091] Single batch magnetic bead stirring and washing steps: control the second pump body P2 to stop; chain control the fifth valve KV-5 to demagnetize and close, the seventh valve KV-7 to close, and the ninth valve KV-9 to close; start the stirring tank R2 and stir for a predetermined time.

[0092] Waste liquid discharge steps: control the first valve KV-1 to close, the second valve KV-2 to open, the fourth valve KV-4 to open, and the sixth valve KV-6 to open; control the magnetic bead separation device M1 to magnetize and start the first pump body P1; obtain the liquid level detection signal of the liquid level sensor in the stirring tank R2, and determine whether the liquid level of the stirring tank R2 is lower than the second preset lower limit height. If not, continue to detect, and if so, proceed to the next step.

[0093] Cleanliness detection step: stop the stirring tank R2, and draw a two-dimensional data graph with respect to time based on the concentration of the target magnetic beads detected by the first magnetic bead sensor AE1 in the waste liquid discharge step; detect whether the peak of the data graph is lower than the second preset magnetic bead concentration. If not, return to the single batch magnetic bead pipetting step; if so, proceed to the next step.

[0094] The steps for pipetting a single batch of finished magnetic bead liquid are as follows: the first pump body P1 is controlled to stop, and the fourth valve KV-4 and the sixth valve KV-6 are controlled to close; the fifth valve KV-5, the seventh valve KV-7 and the eighth valve KV-8 are controlled to open in a chain manner; the magnetic bead separation device M1 is controlled to be demagnetized; the second pump body P2 is controlled to start, and the concentration of the target magnetic beads detected by the second magnetic bead sensor AE2 is detected to be lower than the third preset magnetic bead concentration. If not, the detection is continued; if so, the next step is entered.

[0095] Batch cycle steps: control the fifth valve KV-5 to close, the seventh valve KV-7 to close, and the eighth valve KV-8 to close, control the second pump body P2 to stop, obtain the liquid level detection signal of the liquid level sensor in the raw liquid tank R1, and determine whether the liquid level in the raw liquid tank R1 is lower than the first preset lower limit height. If not, return to the pre-filling step of the magnetic bead separation device.

[0096] Pipeline self-cleaning steps: control the raw liquid tank R1 to stop working, start the mixing tank R2, control the fifth valve KV-5 to open, the seventh valve KV-7 to open, the ninth valve KV-9 to open, and start the second pump body P2; obtain the liquid level detection signal of the liquid level sensor in the mixing tank R2 and determine whether it is higher than the second preset upper limit. When it reaches the second preset upper limit, control the fourth valve KV-4 to open, the sixth valve KV-6 to open, and start the first pump body P1; based on the liquid level detection signal, determine whether the liquid level in the mixing tank R2 is lower than the second preset lower limit. If not, continue to detect. If so, complete the pipeline self-cleaning.

[0097] Equipment shutdown steps: After detecting the production task end signal, the shutdown operation is performed, including controlling the first valve KV-1 to close, the second valve KV-2 to close, the third valve KV-3 to close, the fourth valve KV-4 to close, the fifth valve KV-5 to close, the sixth valve KV-6 to close, the seventh valve KV-7 to close, the eighth valve KV-8 to close, and the ninth valve KV-9 to close in sequence; controlling the first pump body P1 to stop and the second pump body P2 to stop; controlling the raw liquid tank R1 to stop and the stirring kettle R2 to stop; controlling the magnetic bead separation device M1 to perform demagnetization operation to complete the system shutdown and maintain the equipment in a safe static state.

[0098] It should be noted that, in different embodiments, the pre-filling step of the magnetic bead separation device in the above control method of the magnetic bead continuous separation system can be deleted. It plays a role in improving the operation of the entire method rather than a decisive role. Accordingly, the return step in the batch circulation step corresponds to the single batch raw liquid feeding step.

[0099] This application also discloses the following three operating examples of magnetic bead continuous separation systems with different configurations.

[0100] Example 1: Vertical hexagonal magnetic tile and magnetic bead continuous separation system (deionized water washing) Configuration Features 1. Magnetic bead separation device: vertical structure, electric push rod lifting 2. Magnetic device: permanent magnetic hexagonal tiles (magnetic field strength 1.0T) 3. Washing liquid: deionized water 4. Testing instrument: Clarity tester (AE1 / AE2, unit: NTU (scattered turbidity)) 5. Magnetic bead parameters: particle size 1.5μm, material Fe3O4 Key parameter settings Process time statistics: 1. Single washing cycle time: 60 minutes (including steps 4→9) 2. Product purity: 98.9% 3. Product recovery rate: 97.3% Example 2: Horizontal electromagnetic bead continuous separation system (ethanol washing) Configuration Features 1. Magnetic bead separation device: horizontal structure, cylinder lifting 2. Magnetic device: electromagnetic device (maximum magnetic field strength 30T) 3. Washing solution: ethanol (concentration 70%) 4. Detection instrument: spectrophotometer (AE1 / AE2, unit OD600 (600nm wavelength optical density)) 5. Magnetic bead parameters: particle size 60μm, material carboxyl modified Fe3O4 Key parameter settings Parameter name Setting value AE1 high concentration value 0.8OD600 AE1 low concentration value 0.1OD600 AE2 low concentration value 0.05OD600 LIT1 low level value 0.2L LIT2 high level value 100L LIT2 low level value 0.2L R1 stirring speed 50rpm R2 stirring speed 120rpm R2 stirring time 30min P1 speed 80mL / min Process time statistics 1. Single washing cycle time: 120 minutes (including steps 4→9) 2. Product purity: 99.2% 3. Product recovery rate: 97.6% Example 3: Horizontal Permanent Magnetic Diamond Tile Magnetic Bead Continuous Separation System (Polymer Washing) Configuration Features 1. Magnetic bead separation device: horizontal structure, hydraulic transmission lifting 2. Magnetic device: permanent magnetic diamond tiles (magnetic field strength 0.4T) 3. Washing solution: PEG-6000 (molecular weight 6000) solution (concentration 5% w / v (mass concentration)) 4. Testing instrument: turbidity detector (AE1 / AE2, unit FTU (scattered turbidity unit)) 5. Magnetic bead parameters: particle size 0.8μm, material SiO2 coated Fe3O4 Key parameter settings Parameter name Setting value AE1 high concentration value 100FTU AE1 low concentration value 10FTU AE2 low concentration value 2FTU LIT1 low level value 0.5L LIT2 high level value 1000L LIT2 low level value 0.5L R1 stirring speed 50rpm R2 stirring speed 120rpm R2 stirring time 30min P1 speed 80mL / min Process time statistics: 1. Single wash cycle time: 240 minutes (high viscosity wash solution prolongs pipetting time) (including magnetic bead separation device pre-filling → cleanliness test step) 2. Product purity: 96.4% 3. Product recovery rate: 98.5% It can be seen that the continuous automated magnetic bead continuous separation system provided by this application has obvious advantages. The traditional magnetic bead separation process that relies on manual operation has significant economic and efficiency disadvantages. First, in manual repetitive operations, as the scale of processing expands, the required labor costs increase exponentially. For example, taking the processing of 1,000 samples per day as an example, at least 10 operators need to be configured in shifts, each working 8 hours a day, and at 50 yuan / hour, the daily labor cost is 5,000 yuan. Using an automated magnetic bead continuous separation system, only one monitor is required to complete the same workload, and the cost is reduced to about 200 yuan / day, which fundamentally saves manpower expenses. In addition, continuous manual operation also faces problems of operational fatigue and psychological load. The error rate increases significantly from 0.1% to 5% after long-term operation. The resulting sample retesting, repeated reagent input and management burden further push up monthly operating costs, and a conservative estimate of an additional increase of 10,000 yuan.

[0101] In terms of throughput, the processing capacity of manual operations is also difficult to meet the needs of industrial-grade sample processing. Usually, each operator can only process a maximum of 200 samples per day. The overall throughput is limited and difficult to expand. Industrial-grade applications usually need to meet the continuous high-throughput output requirements of tens of thousands of samples per day, and the manual mode is obviously not competent. At the same time, in terms of purity and separation efficiency, the manual mode is prone to uneven distribution of magnetic beads due to the large container size and uncontrollable fluid turbulence, which in turn causes significant fluctuations in the recovery rate of the target object, with a fluctuation range of more than 30%, seriously affecting the consistency and quality stability of the final product.

[0102] In response to the above problems, the continuous automated magnetic bead continuous separation system provided by this application systematically integrates the magnetic bead separation device, stirring tank, automatic valve and fluid control network, and combines online concentration monitoring and liquid level control strategies to build a multi-level chain of automated control logic, which can realize the "raw material in - product out" full-process automated processing path from the feed of raw liquid to the output of magnetic bead liquid finished product. By setting logical linkage between each processing node, real-time control of core parameters such as flow, concentration, temperature, and liquid level can be achieved, effectively avoiding the interference of human intervention on the separation effect, greatly improving processing efficiency and sample consistency, and fundamentally breaking through the throughput bottleneck and precision limitations of manual operation.

[0103] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations on the present application. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present application.

Claims

1. A magnetic bead continuous separation system, characterized in that: include: A magnetic bead separation device M1, wherein the magnetic bead separation device M1 is a hollow tube structure and the tube wall can be magnetized and demagnetized. One end of the magnetic bead separation device M1 is provided with a first liquid inlet (11) and a second liquid inlet (12), and the other end is provided with a first liquid discharge port (13) and a second liquid discharge port (14). The first liquid inlet (11) is away from the tube wall area that can be magnetized and demagnetized relative to the second liquid inlet (12), and the first liquid discharge port (13) is away from the tube wall area that can be magnetized and demagnetized relative to the second liquid discharge port (14); A raw liquid tank R1, the raw liquid tank R1 being connected to the first liquid inlet (11) via a first flow channel (101); A waste liquid tank T3, the waste liquid tank T3 is connected to the first liquid discharge port (13) via a fifth flow channel (105), and a first magnetic bead sensor AE1 is provided on the fifth flow channel (105); A product tank T2, wherein the liquid inlet of the product tank T2 is connected to the second liquid discharge port (14) via a sixth flow channel (106), and a second magnetic bead sensor AE2 is provided on the sixth flow channel (106); A stirred tank R2, wherein the liquid inlet of the stirred tank R2 is connected to the second liquid outlet (14) through a seventh flow channel (107), and the liquid outlet of the stirred tank R2 is connected to the first liquid inlet (11) through a second flow channel (102); A washing liquid tank T1, wherein the liquid outlet of the washing liquid tank T1 is connected to the first liquid inlet (11) through a third flow channel (103), and is connected to the second liquid inlet (12) through a fourth flow channel (104); The first flow channel (101), the second flow channel (102), the third flow channel (103), the fourth flow channel (104), the fifth flow channel (105), the sixth flow channel (106) and the seventh flow channel (107) are all provided with valves, and the first flow channel (101), the second flow channel (102), the third flow channel (103) and the fourth flow channel (104) are provided with pump bodies.

2. The magnetic bead continuous separation system according to claim 1, characterized in that The first flow channel (101) is provided with a first valve KV-1, the second flow channel (102) is provided with a second valve KV-2, and the third flow channel (103) is provided with a third valve KV-3. One end of the first valve KV-1 away from the raw liquid tank R1, one end of the second valve KV-2 away from the stirring tank R2, and one end of the third valve KV-3 away from the washing liquid tank T1 are connected to one end of the fourth valve KV-4 through the first pump body P1, and the other end of the fourth valve KV-4 is connected to the first liquid inlet. (11); the fourth flow channel (104) is provided with a second pump body P2 and a fifth valve KV-5; the fifth flow channel (105) is provided with a flow sensor and a sixth valve KV-6; the sixth flow channel (106) is provided with an eighth valve KV-8, and the seventh flow channel (107) is provided with a ninth valve KV-9, and the end of the eighth valve KV-8 away from the product tank T2 and the end of the ninth valve KV-9 away from the waste liquid tank T3 are connected to the second liquid discharge port (14) through the seventh valve KV-7.

3. The magnetic bead continuous separation system according to claim 1, characterized in that A liquid level sensor is provided in the stirring tank R2, and the liquid level sensor is correspondingly set with a second preset upper limit height and a second preset lower limit height.

4. The magnetic bead continuous separation system according to claim 1, characterized in that The magnetic bead separation device M1 is used to separate the raw liquid to be separated containing target magnetic beads, which are particles that can be effectively captured by the external magnetic field, and the non-magnetic components in the non-magnetic waste liquid are impurity components that are not captured by the external magnetic field.

5. The magnetic bead continuous separation system according to claim 1, characterized in that The magnetic bead separation device M1 comprises a magnetic bead separator (1), a magnetic field generating device (2) and a driving device (3), wherein the driving device (3) is used to drive the magnetic field generating device (2) to move closer to or away from the side wall of the magnetic bead separator (1), the first liquid inlet (11) and the second liquid inlet (12) are arranged at one end of the magnetic bead separator (1), and the first liquid discharge port (13) and the second liquid discharge port (14) are arranged at the other end of the magnetic bead separator (1).

6. A method for controlling a continuous magnetic bead separation system, for use in the continuous magnetic bead separation system according to any one of claims 1 to 5, comprising the following steps: Single batch stock solution feeding step: Control the third valve KV-3 to close, the first valve KV-1 to open, and detect whether the concentration of target magnetic beads detected by the first magnetic bead sensor AE1 reaches the first preset magnetic bead concentration. If not, continue the detection; if so, proceed to the next step; the target magnetic beads are particles that can be effectively captured by the magnetic field applied by the magnetic bead separation device M1; Single-batch magnetic bead pipetting steps: Control the first pump body P1 to stop, the fourth valve KV-4 to close, and the sixth valve KV-6 to close; interlock control the fifth valve KV-5, the seventh valve KV-7, and the ninth valve KV-9 to open, and control the magnetic bead separation device M1 to demagnetize; interlock control the second pump body P2 to start, and start the stirring tank R2; obtain the liquid level detection signal of the liquid level sensor in the stirring tank R2, and determine whether the liquid level of the stirring tank R2 is higher than the second preset upper limit. If not, continue to detect; if so, proceed to the next step; Single batch magnetic bead stirring and washing steps: control the second pump body P2 to stop; chain control the fifth valve KV-5 to close, the seventh valve KV-7 to close, and the ninth valve KV-9 to close; start the stirring tank R2 and stir for a predetermined time; Waste liquid discharge step: control the first valve KV-1 to close, the second valve KV-2 to open, the fourth valve KV-4 to open, and the sixth valve KV-6 to open; control the magnetic bead separation device M1 to magnetize and start the first pump body P1; obtain the liquid level detection signal of the liquid level sensor in the stirring tank R2, and determine whether the liquid level of the stirring tank R2 is lower than the second preset lower limit. If not, continue the detection; if so, proceed to the next step; Cleanliness detection step: Stop the stirred tank R2 and draw a two-dimensional data graph with respect to time based on the concentration of the target magnetic beads detected by the first magnetic bead sensor AE1 in the waste liquid discharge step; detect whether the peak of the data graph is lower than the second preset magnetic bead concentration. If not, return to the single batch magnetic bead pipetting step; if so, proceed to the next step; Single batch of finished magnetic bead liquid pipetting steps: control the first pump body P1 to stop, control the fourth valve KV-4 and the sixth valve KV-6 to close; control the fifth valve KV-5, the seventh valve KV-7 and the eighth valve KV-8 to open; control the magnetic bead separation device M1 to demagnetize; control the second pump body P2 to start, and detect whether the concentration of target magnetic beads detected by the second magnetic bead sensor AE2 is lower than the third preset magnetic bead concentration. If not, continue the detection; if so, proceed to the next step; Batch cycle steps: control the fifth valve KV-5 to close, the seventh valve KV-7 to close, and the eighth valve KV-8 to close, control the second pump body P2 to stop, obtain the liquid level detection signal of the liquid level sensor in the raw liquid tank R1, and determine whether the liquid level in the raw liquid tank R1 is lower than the first preset lower limit height. If not, return to the single batch raw liquid feeding step.

7. The control method of the magnetic bead continuous separation system according to claim 6, characterized in that: The following steps are also included: Pipeline self-cleaning steps: control the raw liquid tank R1 to stop working, start the stirring kettle R2, control the fifth valve KV-5 to open, the seventh valve KV-7 to open, the ninth valve KV-9 to open, and start the second pump body P2; obtain the liquid level detection signal of the liquid level sensor in the stirring kettle R2, and judge whether the liquid level in the stirring kettle R2 is higher than the second preset upper limit. When it reaches the second preset upper limit, control the fourth valve KV-4 to open, the sixth valve KV-6 to open, and start the first pump body P1; based on the liquid level detection signal, judge whether the liquid level is lower than the second preset lower limit. If not, continue to detect. If so, complete the pipeline self-cleaning.

8. The control method of the magnetic bead continuous separation system according to claim 7, characterized in that: The following steps are also included: Pre-setting step, setting the first set value, the second set value, the third set value; setting the first preset lower limit height, the second preset upper limit height and the second preset lower limit height; setting the stirring speed and stirring time of the raw liquid tank R1 and the stirring kettle R2.

9. The control method of the magnetic bead continuous separation system according to claim 7, characterized in that: Before the single batch stock solution feeding step, the following steps are also included: Magnetic bead separator pre-filling steps: control the second valve KV-2 to close, the third valve KV-3 to open, the fourth valve KV-4 to open, and the sixth valve KV-6 to open, control the magnetic bead separator M1 to magnetize and then start the first pump body P1; check whether the flow sensor detects the flow passing, if not, continue to test, if yes, go to the next step.

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