A magnetic bead continuous separation system and control method

Abstract

The application relates to the technical field of nanometer 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 in a hollow tube structure and can be magnetized and demagnetized at the tube wall, one end of the magnetic bead separation device is provided with a first liquid inlet and a second liquid inlet, the other end is provided with a first liquid outlet and a second liquid outlet, a raw liquid tank is connected to the first liquid inlet through a first flow channel, a waste liquid tank is connected to the first liquid outlet through a fifth flow channel, the fifth flow channel is provided with a first magnetic bead sensor, a product tank is connected to the second liquid outlet through a sixth flow channel, the sixth flow channel is provided with a second magnetic bead sensor, a 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 the second flow channel; a washing liquid tank is connected to the first liquid inlet through a third flow channel and connected to the second liquid inlet through a fourth flow channel. The system has the advantages of meeting the requirements of continuous separation and purification of magnetic beads in an automatic and unlimited batch mode.

Description

TECHNICAL FIELD

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

[0002] Magnetic beads are micron or nanometer particle structures with good magnetic response characteristics. The core is usually composed of magnetic inorganic materials (such as ferroferric oxide), the outer layer is coated with polymer or siliceous material, and specific functional groups such as carboxyl or amino are introduced on the surface, thereby having the ability to specifically bind to target molecules. Due to the magnetic control operability and high flexibility of functional modification, magnetic beads are widely used in nucleic acid extraction, protein purification, cell sorting, immunoassay and magnetic control drug delivery in many biological and industrial fields. After synthesis, magnetic beads often have problems such as uneven magnetism, impurity residues and inconsistent particle size distribution. If not purified and selected, it will directly affect the magnetic response and binding specificity, and then reduce the separation efficiency and repeated use performance in subsequent applications. Therefore, after the preparation of magnetic beads is completed, it is usually necessary to separate and enrich them to remove particles with weak magnetism, abnormal size or high impurity content, and 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. The common method includes dispensing the magnetic bead suspension into centrifuge tubes, realizing magnetic attraction and discarding liquid on the fixed magnetic stand by manual operation, and then completing rough screening through multiple washing operations. Each link is usually completed in different centrifuge tubes, magnetic stands or other containers in sequence, which not only has a complicated process and relies on manual operation, but also is easy to cause magnetic bead loss, captured material residues and cross contamination in the transfer process. In addition, due to the different standards for determining the washing endpoint by manual judgment, the product purity and recovery rate often fluctuate, which is difficult to meet the needs of large-scale and high-consistency industrial production. SUMMARY

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

[0005] In a first aspect, the application provides a magnetic bead continuous separation system, which adopts the following technical scheme:

[0006] A magnetic bead continuous separation system comprises:

[0007] The magnetic bead separation device is a hollow tube structure, and the tube wall 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 is provided with a first liquid outlet and a second liquid outlet. The first liquid inlet is away from the tube wall area that can be magnetized and demagnetized relative to the second liquid inlet. The first liquid outlet is away from the tube wall area that can be magnetized and demagnetized relative to the second liquid outlet.

[0008] The raw liquid tank is connected to the first liquid inlet through a first flow channel.

[0009] The waste liquid tank is connected to the first liquid outlet through a fifth flow channel, and the fifth flow channel is provided with a first magnetic bead sensor. The product tank is connected to the second liquid outlet through a sixth flow channel, and the sixth flow channel is provided with a second magnetic bead sensor.

[0010] The stirring kettle is connected to the second liquid outlet through a seventh flow channel, and the outlet of the stirring kettle is connected to the first liquid inlet through a second flow channel.

[0011] The washing liquid tank is connected to the first liquid inlet through a third flow channel and connected to the second liquid inlet through a fourth flow channel.

[0012] 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.

[0013] Optionally, the first flow channel is provided with a first valve, the second flow channel is provided with a second valve, the third flow channel is provided with a third valve, and the end of the first valve away from the raw liquid tank, the end of the second valve away from the stirring kettle and the end of the third valve away from the washing liquid tank are connected to one end of a fourth valve through a first pump body. The other end of the fourth valve is connected to the first liquid inlet. The fourth flow channel is provided with a second pump body and a fifth valve. The fifth flow channel is provided with a flow sensor and a sixth valve. The sixth flow channel is provided with an eighth valve, and the seventh flow channel is provided with a ninth valve. The end of the eighth valve away from the product tank and the end of the ninth valve away from the waste liquid tank are connected to the second liquid outlet through a seventh valve.

[0014] Optionally, a liquid level sensor is arranged in the stirring kettle, and the liquid level sensor is correspondingly provided with a second preset upper limit and a second preset lower limit.

[0015] Optionally, the magnetic bead separation device is used for separating the target magnetic beads in the raw liquid to be separated. The target magnetic beads are particles that can be effectively captured by an 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.

[0016] In a second aspect, the application provides a control method of a magnetic bead continuous separation system, which adopts the following technical scheme: a control method of a magnetic bead continuous separation system, for a magnetic bead continuous separation system, comprising the following steps:

[0017] Single batch of raw liquid feeding step: control the third valve to be closed and the first valve to be opened, and detect whether the concentration of the target magnetic beads detected by the first magnetic bead sensor reaches the first preset magnetic bead concentration, if not, continue to detect, if yes, proceed to the next step; wherein the target magnetic beads are particles that can be effectively captured by the magnetic field of the magnetic bead separation device;

[0018] Single batch of magnetic bead pipetting step: control the first pump body to stop, the fourth valve to be closed, and the sixth valve to be closed; control the fifth valve, the seventh valve, and the ninth valve to be opened in a chain control manner, and control the magnetic bead separation device to be demagnetized; control the second pump body to be started in a chain control manner, and control the stirring tank to be started; acquire 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 the second preset upper limit, if not, continue to detect, if yes, proceed to the next step;

[0019] Single batch of magnetic bead stirring and washing step: control the second pump body to stop; control the fifth valve, the seventh valve, and the ninth valve to be closed in a chain control manner; start the stirring tank and stir for a predetermined time;

[0020] Waste liquid discharge step: control the first valve to be closed, the second valve to be opened, the fourth valve to be opened, and the sixth valve to be opened; control the magnetic bead separation device to be magnetized, and control the first pump body to be started; acquire 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 lower than the second preset lower limit, if not, continue to detect, 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 the target magnetic beads detected by the first magnetic bead sensor in the waste liquid discharge step; detect whether the peak value of the data graph is lower than the second preset magnetic bead concentration, if not, return to the single batch of magnetic bead pipetting step, if yes, proceed to the next step;

[0021] Single batch of magnetic bead liquid product pipetting step: control the first pump body to stop, and control the fourth valve and the sixth valve to be closed; control the fifth valve, the seventh valve, and the eighth valve to be opened in a chain control manner; control the magnetic bead separation device to be demagnetized; control the second pump body to be started, and detect whether the concentration of the target magnetic beads detected by the second magnetic bead sensor is lower than the third preset magnetic bead concentration, if not, continue to detect, if yes, proceed to the next step;

[0022] Batch cycle step: control the fifth valve, the seventh valve, and the eighth valve to be closed, and control the second pump body to stop; acquire 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, if not, return to the single batch of raw liquid feeding step.

[0023] Optionally, the method further comprises the following steps:

[0024] The pipeline self-cleaning step: control the raw liquid tank to stop working, start the stirring tank, control the fifth valve to open, the seventh valve to open, the ninth valve to open, and start the second pump body; obtain the liquid level detection signal of the liquid level height sensor in the stirring tank and judge whether it is higher than the second preset upper limit, until it is reached, then control the fourth valve to open, the sixth valve to open, and the first pump body to start; based on the liquid level detection signal, judge whether the liquid level height in the stirring tank is lower than the second preset lower limit height, if not, continue to detect, if yes, complete the pipeline self-cleaning.

[0025] Optionally, the method further comprises the following steps:

[0026] The preset step sets the first set value, the second set value, and the third set value; sets the first preset lower limit height, the second preset upper limit height, and the second preset lower limit height; and sets the stirring speed and stirring time of the raw liquid tank and the stirring tank.

[0027] Optionally, before the single-batch raw liquid feeding step, the method further comprises the following steps:

[0028] The magnetic bead separator pre-liquid step: 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 add magnet after starting the first pump body P1; detect whether the flow sensor detects the flow, if not, continue to detect, if yes, go to the next step.

[0029] In summary, the present application includes at least one of the following beneficial technical effects:

[0030] 1. Realize the continuous, automatic and unlimited batch separation operation of magnetic beads, break through the problems of process fragmentation, large manual operation error, limited system flux, and parameter dependent manual adjustment in traditional magnetic bead separation technology; the device can complete the whole process automatic control from raw material input, magnetic separation, washing and flushing to the separation and recovery of target magnetic bead liquid, significantly improving the efficiency and stability of the magnetic bead separation process.

[0031] 2. The continuous automatic magnetic bead separation system adopts a multi-parameter adjustable structure and supports modular magnetic component configuration, so that users can flexibly adapt to different particle sizes and different magnetic response characteristics of magnetic bead types without replacing the main equipment, significantly improving the adaptability and processing scale expansion ability of the device, and greatly reducing the labor cost.

[0032] 3. By introducing liquid level control, online detection and valve interlocking logic in the washing process, the system can realize automatic determination and precise control of the washing steps, reduce over-washing or under-washing phenomenon, ensure the recovery rate and purity of the target object in the optimal interval, and significantly improve the batch consistency and repeatability of the magnetic bead separation product. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 is a pipeline connection diagram of a magnetic bead continuous separation system in an embodiment of the present application.

[0034] Figure 2 is a structural schematic diagram of a magnetic bead separation device in an embodiment of the present application.

[0035] Figure 3 is a whole schematic diagram of a magnetic field generating device in an embodiment of the present application Figure 1 .

[0036] Figure 4 is a whole schematic diagram of a magnetic field generating device in an embodiment of the present application Figure 2 .

[0037] Figure 5 is a flow chart of a control method of a magnetic bead continuous separation system in an embodiment of the present application Figure 1 .

[0038] Figure 6 is a flow chart of a control method of a magnetic bead continuous separation system in an embodiment of the present application Figure 2 .

[0039] Figure 7 is a flow chart of a control method of a magnetic bead continuous separation system in an embodiment of the present application Figure 3 .

[0040] Reference signs:

[0041] 1, magnetic bead separator; 11, first liquid inlet; 12, second liquid inlet; 13, first liquid outlet; 14, second liquid outlet;

[0042] 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;

[0043] 2, magnetic field generating device; 21, anti-exfoliation layer; 22, embedded magnetic layer; 221, support plate; 222, magnetic tile; 223, limiting groove; 23, shielding layer;

[0044] 3, driving device; 31, motor module; 32, support rod; 33, transmission rod;

[0045] 4, support; 5, support column; 6, sliding structure. DETAILED DESCRIPTION

[0046] Embodiments of the present application are described in detail below, examples of which are shown in the accompanying drawings.

[0047] In the description of the present specification, the description of the terms "certain embodiments", "one embodiment", "some embodiments", "illustrative embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials or characteristics described in connection with the described embodiments or examples are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Also, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0048] Magnetic beads are a kind of magnetic microspheres, whose core is usually composed of inorganic materials with magnetism (such as ferroferric oxide), and the shell is wrapped with high polymer materials or siliceous materials. By introducing functional groups such as carboxyl, amino, streptavidin, etc. on the surface, the magnetic beads can have the ability to specifically bind biological molecules, and are widely used in the fields of biomedicine and industry such as nucleic acid extraction, protein purification, cell sorting, immune detection and magnetic controlled drug delivery. The size of magnetic beads is generally between nanometer and micrometer, and they have good magnetic response, which can quickly aggregate under the action of an external magnetic field, realizing efficient capture and transfer of target objects.

[0049] However, after the synthesis of magnetic beads is completed, there is often a phenomenon of uneven magnetism in the system, that is, there is a difference in the magnetic response strength between different particles. At the same time, some magnetic beads may be attached with incomplete removal of reaction by-products, free monomers or inorganic impurities, which will affect the separation efficiency and binding specificity of the magnetic beads in actual application. Therefore, strict separation and screening operation is needed to eliminate particles with weak magnetism, high impurity content or size deviating from the set range.

[0050] In this process, the magnetic beads with different magnetic strengths can be effectively separated and 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 then place them on a fixed magnetic stand. Particles with sufficient magnetic response are captured on the tube wall by the magnetic field, and the remaining non-magnetic or low-magnetic impurities are discarded with the supernatant. Then, the remaining magnetic beads are resuspended with liquid, and the rough enrichment and purification are realized through multiple "capture-liquid discard-liquid addition-mixing" operations. If a magnetic rod type device is used, manual or mechanical arm cooperation is needed to immerse the magnetic rod into the magnetic bead suspension, capture it, and then transfer it to the washing liquid to complete further washing.

[0051] However, the above-mentioned traditional magnetic bead purification operation has obvious limitations. First, its operation process presents a highly fragmented feature, and each link often depends on manual step-by-step completion, which can easily lead to increased magnetic bead loss, reduced processing efficiency, and heavy burden on the operator, and the single batch processing volume is also small.

[0052] In practical applications, although the existing magnetic bead continuous separation system can cover the needs of part of the standard process, the overall design generally lacks systematic consideration of processing flexibility and scalability, especially when dealing with magnetic beads of different magnetic response levels or varying single batch processing volumes. For example, when dealing with micro-magnetic particles or expanding the batch volume, the existing system can usually only be dealt with by replacing the magnet with a stronger one or increasing the device volume, which leads to complex device structure, increased system cost and decreased running stability. In addition, since the magnetic attraction efficiency is closely related to the magnetic field gradient distribution, blindly increasing the magnetic strength often cannot effectively solve the problem of poor magnetic bead selectivity, but may cause excessive enrichment of non-target particles.

[0053] Further, the lack of automation in the washing process is one of the key pain points that the existing technology needs to solve. Traditional magnetic bead separation equipment often relies on manual liquid addition, mixing and capture processes in the washing operation, lacks reliable termination criteria, and especially when removing non-specific binding components, repeated washing is needed to ensure complete removal of residual impurities. In this process, the operator needs to judge whether the washing is sufficient based on experience, which is easy to cause excessive washing or insufficient washing due to judgment deviation, thereby affecting the purity of the magnetic beads.

[0054] In order to solve the problems of low separation efficiency, unstable washing process and insufficient automation in the existing magnetic bead separation process, the embodiments of the present application provide a magnetic bead continuous separation system. The overall structure of the system is shown in Figure 1 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, each component is connected to each other through multiple flow channels, and is equipped with a valve assembly for controlling process switching.

[0055] The magnetic bead separation device M1 contains a hollow flow channel, and the pipe wall region of the flow channel can stably provide and eliminate a controllable magnetic field to achieve 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 outlet 13 and a second liquid outlet 14 at the other end. The first liquid inlet 11 and the first liquid outlet 13 are respectively located on the side away from the magnetic field action region, and the second liquid inlet 12 and the second liquid outlet 14 are located on the side close to the magnetic field action region relative to the first liquid inlet 11 and the first liquid outlet 13. Different inlet and outlet selection and combination are performed through the first liquid outlet 13, the second liquid outlet 14, the second liquid inlet 12 and the second liquid outlet 14, so that the liquid containing the magnetic beads to be processed is circulated in the same magnetic bead separation device M1, and different purposes such as capture, separation and washing are achieved in different stages. The raw liquid tank R1 is used to store the raw liquid containing the 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.

[0056] The waste liquid tank T3 is connected to the first liquid outlet 13 of the magnetic bead separation device M1 through the fifth flow channel 105, and the first magnetic bead sensor AE1 is arranged on the flow channel to detect the residual magnetic bead concentration in the waste liquid in real time. The product tank T2 is connected to the second liquid outlet 14 through the sixth flow channel 106, and the second magnetic bead sensor AE2 is arranged on the flow channel to monitor the concentration of the target magnetic beads in the finished product liquid.

[0057] The stirring tank R2 is an execution device of the resuspension link, and the inlet is connected to the second liquid outlet 14 through the seventh flow channel 107 to receive the target magnetic bead liquid after separation and purification; and the outlet is connected back to the first liquid inlet 11 through the second flow channel 102 to realize closed-loop washing operation of the target magnetic beads. The washing tank T1 is connected to the first liquid inlet 11 through the third flow channel 103 and connected to the second liquid inlet 12 through the fourth flow channel 104 to provide the system with the ability to inject washing liquid through different paths.

[0058] In order to accurately control the fluid transmission process, independent valves are arranged on the first to seventh flow channels, and pump bodies are arranged on the first to fourth flow channels to switch and control the opening and closing state of the liquid path and the liquid flow in each operation stage.

[0059] Firstly, the magnetic bead separation device M1 arranged in the system adopts an adjustable modular magnetic structure, which can be flexibly configured according to the processing volume in different processing tasks. By increasing or decreasing the magnetic modules or adjusting the magnetic field distribution length, the processing capacity of each batch in the processing task can be expanded or contracted, which can adapt to the needs of multiple scenes from small-scale experiments to large-scale preparation.

[0060] Secondly, the degree of automation of the washing process is greatly improved. The liquid level height of the stirring tank R2 in the system is monitored in real time by a liquid level sensor, which is used as a criterion for starting and stopping the washing cycle. The system realizes automatic closed-loop control of the whole process from liquid transfer, magnetic bead resuspension to washing liquid replacement, significantly reducing the dependence on manual operation. In combination with the online detection unit, such as conductivity sensor or fluorescent probe, deployed at the outlet end of the washing device, the system can evaluate the impurity concentration in the washing liquid in real time, and adjust the three-way valve to control the washing cycle and the washing liquid volume, thereby realizing quantitative management and standardized scheduling of the washing process. The control accuracy of the washing parameters is high, and the coefficient of variation in the system can be controlled below 1%. Compared with the fluctuation amplitude of more than 15% in traditional manual operation, the consistency and process stability between batches are significantly enhanced, meeting the quality system requirements of GMP and GLP in the field of drug production.

[0061] Specifically, referring to Figure 2 , the magnetic bead separation device M1 includes a magnetic bead separator 1, a magnetic field generating device 2, and a driving device 3, which cooperatively form a closed separation channel in a controllable magnetic field environment, capable of realizing automatic capture, separation and washing of target magnetic beads in the original liquid to be separated. The target magnetic beads are particles that can be effectively captured by an external magnetic field, and the non-magnetic components in the non-magnetic waste liquid are impurity components that are not captured by an external magnetic field.

[0062] Specifically, the magnetic bead separator 1 is a hollow structure for processing the original liquid containing target magnetic beads. The magnetic field generating device 2 is arranged outside 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 pipe wall. The driving device 3 is linked with the magnetic field generating device 2, which is used to adjust the distance between the magnetic field generating device 2 and the side wall of the magnetic bead separator 1, so as to change the magnetic field strength in the magnetic bead separator 1. In different embodiments, the magnetic bead separator 1 can be fixed by different forms of supports 4, and the form of the support 4 can be adjusted according to the arrangement of the magnetic bead separator 1. When the magnetic bead separator 1 is horizontal or vertical, the support 4 should be adjusted accordingly, which is not limited in this application.

[0063] Referring to Figure 2 , one end of the magnetic bead separator 1 is provided with a first liquid inlet 11 and a second liquid inlet 12, and the other end is provided with a first liquid outlet 13 and a second liquid outlet 14. The first liquid inlet 11 and the second liquid inlet 12 are used to introduce the original liquid containing target magnetic beads and the washing liquid, realizing step-by-step alternating liquid input. The first liquid outlet 13 is used to discharge the impurities contained in the original liquid and the suspended waste liquid, and the second liquid outlet 14 is used to guide the magnetic bead liquid containing target magnetic beads. With the above interface configuration, the whole process of original liquid processing, magnetic bead capture and impurity flushing can be completed in a single cavity structure.

[0064] In various embodiments of the present application, the magnetic bead separator 1 can be arranged in various orientations with respect to the horizontal plane. The magnetic bead separator 1 can be arranged in a vertical orientation, a horizontal orientation, or an inclined orientation. The vertical orientation can also be referred to as a standing orientation, and the horizontal orientation can also be referred to as a lying orientation.

[0065] In an exemplary embodiment, the magnetic bead separator 1 is arranged in a horizontal orientation, and the magnetic field generating device 2 is attached to the outer side of the tube. In this arrangement, the first inlet 11 and the second inlet 12 are arranged at one end of the magnetic bead separator 1, and the first outlet 13 and the second outlet 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 lumen, the capture of the target magnetic beads can be continuously achieved. Further, the second inlet 12 is arranged on the side of the magnetic bead separator 1 closer to the magnetic field generating device 2 relative to the first inlet 11, and the second outlet 14 is arranged on the side of the end of the magnetic bead separator 1 closer to the magnetic field generating device 2 relative to the first outlet 13.

[0066] In a preferred embodiment, the edge of the second inlet 12 is tangent to the wall of the magnetic bead separator 1, so that the injected liquid can flow along the axial direction of the magnetic bead separator 1. The second outlet 14 is also arranged in a similar manner to be tangent to the wall of the magnetic bead separator 1, so as to guide the target magnetic beads to be discharged in the axial direction. The advantage of this arrangement is that during the injection of the washing liquid, an axial impact is quickly formed through the second inlet 12, so that the deposited target magnetic beads can be efficiently flushed to the second outlet 14.

[0067] In a preferred embodiment of the present application, the magnetic field generating device 2 is arranged outside the magnetic bead separator 1, and the structure and arrangement of the magnets are designed to generate a magnetic field in the magnetic bead separator 1 that gradually weakens from the wall to the middle of the tube.

[0068] 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 in the axial direction of the magnetic bead separator 1. The two ends of the magnetic bead separation zone are connected to the first inlet 11 and the first outlet 13, and 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 connected to the second inlet 12 and the second outlet 14, respectively, for injecting the washing liquid and outputting the magnetic bead liquid. 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 towards the magnetic bead capture zone; and the magnetic bead capture zone is close to the magnetic field action region and has a higher magnetic field strength, which can capture and fix the migrated magnetic beads.

[0069] It should be noted that the size of the first liquid inlet 11 and the first liquid outlet 13 does not strictly correspond to the width of the magnetic bead separation zone, and the size of the second liquid inlet 12 and the second liquid outlet 14 does not strictly correspond to the width of the magnetic bead capture zone. The magnetic bead separation zone and the magnetic bead capture zone do not actually exist, but are functional divisions of the interior of the magnetic bead separator 1 according to the behavior of the magnetic beads in the magnetic bead separation liquid.

[0070] During operation, the target magnetic beads in the liquid to be separated maintain a certain spatial dispersion between particles in the magnetic bead separation zone and do not form agglomeration or deposition, but gradually move towards the magnetic bead capture zone under the action of the magnetic field. As it enters the magnetic bead capture zone, the magnetic beads are quickly captured to the surface of the tube wall near the magnetic field generating device and are compressed on the tube wall to form a layer of densely arranged magnetic bead layer, and the magnetic bead particles are fixed to each other by the magnetic field force. In the operation of removing the magnetic field, the magnetic field strength is rapidly weakened, and the capture force between the particles is suddenly reduced to an almost disappeared state, at which point only a certain volume of washing liquid needs to be injected to quickly detach the magnetic beads on the tube wall and bring them into the fluid using the impact force of the liquid.

[0071] In different embodiments, the magnetic field generating device 2 and the driving device 3 can be arranged in one group to achieve partial coverage of the magnetic bead separator 1, or arranged in multiple groups to surround the magnetic bead separator 1 in a manner to increase the effective area of the magnetic field action region. In specific implementation, multiple groups of magnetic field generating devices 2 can be distributed in sequence around the circumference of the magnetic bead separator 1, or arranged in sequence along the axial direction of the magnetic bead separator 1, or arrayed on the surface of the magnetic bead separator 1, forming a composite magnetic field system with directionality and gradient characteristics.

[0072] Specifically, in one embodiment, the magnetic field generating device 2 adopts a full-coverage configuration, that is, through the combination of two or more groups of magnetic field generating devices 2 and driving devices 3, the magnetic field generating device 2 forms a closed ring structure along the outer wall of the magnetic bead separator 1, fully covering the entire circumferential surface of the magnetic bead separator 1. This full-coverage arrangement can build a higher strength and more uniform magnetic field in the magnetic bead separator 1, ensuring that target magnetic beads at any position in the tube cavity are effectively attracted.

[0073] Correspondingly, in another easy-to-assemble embodiment, the magnetic field generating device 2 can adopt a half-coverage configuration, that is, only one or more groups of magnetic field generating devices 2 are arranged on part of the outer wall of the magnetic bead separator 1, so that it forms an effective magnetic field capture region while leaving part of the space for device observation or other functional module expansion.

[0074] In different embodiments, the magnetic field generating device 2 can adopt the principle of electromagnet to generate a magnetic field in the magnetic bead separation device 1, or adopt 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 structure of the permanent magnet, and the magnetic field generating device 2 is described with reference to Figure 3 In the embodiments of the present application, the magnetic field generating device 2 comprises an anti-falling layer 21, an embedded magnetic layer 22 and a shielding layer 23 arranged in sequence from inside to outside, and the three-layer structure is used to establish a suitable magnetic field environment in the magnetic bead separator 1 to achieve the capture and release of the target magnetic beads.

[0075] The first layer is the anti-falling layer 21, that is, the inner layer fitting 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 tightly contact with the outer wall of the magnetic bead separator 1. The anti-falling 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 to prevent the magnetic tiles 222 from displacement or falling off, and on the other hand, it will not significantly block or weaken the magnetic induction lines, ensuring that the magnetic field can effectively penetrate into the inside of the separation tube. In other embodiments, the anti-falling layer 21 can also be made of non-magnetic titanium alloy, engineering plastic or aluminum alloy, as long as it can have good mechanical strength, wear resistance and magnetic field penetration performance. The geometric profile of the anti-falling layer 21 is adapted to the shape of the outer wall surface of the magnetic bead separator 1 to achieve a tight covering effect.

[0076] It should be noted that in some other embodiments, when the embedded magnetic layer 22 can stably fix the magnetic tiles 222 alone, the anti-falling layer 21 can be removed, that is, the magnetic field generating device 2 comprises a two-layer structure of the embedded magnetic layer 22 and the shielding layer 23 arranged in sequence from inside to outside.

[0077] The second layer is the embedded magnetic layer 22, which is the core functional layer of the magnetic field generating device 2, and a plurality of groups of magnetic tiles 222 are embedded therein. The common materials of the magnetic tiles 222 include neodymium iron boron, samarium cobalt or other high-performance rare earth magnetic materials. The embedded magnetic layer 22 is also made of stainless steel material to take into account the structure support and magnetic field transmission characteristics. Correspondingly, the structure form of the embedded magnetic layer 22 can be divided into two types of half-enclosed type and full-enclosed type according to the magnetic field distribution requirements. The full-enclosed structure covers the magnetic bead separation device by a continuous ring-shaped magnet module, which can significantly improve the capture area.

[0078] The third layer is the shielding layer 23, which is the outermost protective layer of the magnetic field generating device 2, and is used to shield the external leakage magnetic field. The shielding layer 23 can limit the magnetic induction lines from spreading to the area outside the embedded magnetic layer 22, preventing the magnetic field from interfering with the surrounding electronic elements, signal lines or operating equipment. At the same time, the shielding layer 23 also has a certain protection strength, which can resist external mechanical impact.

[0079] Referring to Figure 4 In a further embodiment of the present application, the embedded magnetic layer 22 comprises a support plate 221 and a plurality of magnetic tiles 222, the support plate 221 being used to carry and fix the magnetic tiles 222. The support plate 221 is integrally connected or separately connected with the shielding layer 23, and a plurality of limiting grooves 223 for limiting the movement of the magnetic tiles 222 are formed on the support plate 221. Each magnetic tile 222 is fixed by being embedded in the corresponding limiting groove 223, thereby ensuring that the magnetic tiles 222 will not deviate or loosen during the operation of the device.

[0080] In an optional embodiment of the present application, referring to Figure 2 , the driving device 3 comprises 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 internally provided with the transmission rod 33. The motor module 31 outputs through the transmission rod 33 and cooperates with the transmission rod 33 in the bracket 4 through a bevel gear, the rotation of the transmission rod 33 drives the sliding structure 6 connected with it by screwing to move upward, thereby pushing the magnetic field generating device 2 to move up and down. In different embodiments, the motor module can be a pneumatic cylinder, an electric push rod or other different forms. In addition, the transmission form of the transmission rod 33 here can also be embodied in different forms, such as equivalent replacement of worm gear and worm cooperation transmission, or equivalent replacement of gear and rack cooperation transmission, or equivalent replacement of hydraulic transmission, screw transmission, etc. The motor module 31 is installed on a support column located in the middle of the bracket 4, the support rod 32 is arranged at the top end of the support column and is slidably connected with the top end of the support column in the axial direction of the support column, which plays a directional role for the translation of the magnetic field generating device 2.

[0081] In summary, the magnetic bead separation device M1 adopts a fully closed structure, the whole cavity is not provided with traditional stirring shaft, blade and other rotating movement components, and the internal space form design has no dead angle, which effectively avoids the problem of magnetic bead retention or accumulation caused by stirring mechanism, and improves the separation efficiency and sample recovery rate. In the prior art, many magnetic bead processing devices with inclined insertion pipe design cause the magnetic beads to be retained in the separation cavity due to the frequent interference of the stirring components with the magnetic bead deposition area, thereby causing the yield to decrease or the separation to fail. The present application simplifies the inner cavity structure and cancels the mechanical stirring device, which significantly reduces the interference with the target magnetic beads while maintaining the uniform distribution and sufficient contact of the magnetic beads with the magnetic field.

[0082] In the magnetic bead continuous separation system described in the present application, the stock tank R1 serves as a storage unit for the liquid to be treated, and its structure should meet the requirements of storage stability and continuous liquid supply for large quantities of magnetic bead raw materials. The stock tank R1 is in communication with the first liquid inlet 11 of the magnetic bead separation device M1 through the first flow channel 101, and is used to deliver the pre-configured magnetic bead-containing stock solution to the magnetic bead separation device M1 as needed for subsequent capture and separation of target magnetic beads.

[0083] Specifically, the raw liquid tank R1 is a non-process type liquid storage container, and the whole adopts a double-elliptical head structure. Compared with a conventional flat-bottom or single-elliptical structure tank body, the double-elliptical structure has a larger total volume under the condition of the same diameter, and is particularly suitable for long-term storage of large batches of porcelain bead separation liquid. In the internal structure, the raw liquid tank R1 is equipped with multiple sets of stirring paddle devices driven by an external motor, which can be started and stopped through a “one-key start” signal and system interlocking, to realize periodic stirring operation of the magnetic bead raw liquid in the tank, prevent the magnetic beads from settling and gathering after long-term standing, and thus ensure that the raw liquid to be processed in the magnetic bead separation device M1 always maintains a state of uniform particle distribution.

[0084] In different embodiments, the raw liquid tank R1 can be a single-layer wall structure or a double-layer jacket structure for forming an adjustable temperature control system. Temperature adjusting medium is introduced into the jacket through a heating and refrigeration circulating machine to realize real-time adjustment and control of the temperature of the magnetic bead raw liquid in the tank. To further ensure the accuracy of temperature control, a temperature measuring instrument fixedly connected to the tank wall is arranged in the tank body, and the measurement result forms a closed-loop linkage with the external temperature control system to ensure that the temperature in the tank always maintains within the set range.

[0085] In addition, to facilitate system cleaning operation, the raw liquid tank R1 is provided with a spray port on the top, which is designed in cooperation with the position and angle of the tank wall to cover most of the surface area in the tank, so as to effectively flush the liquid hanging on the tank wall or the magnetic bead residues. A liquid level meter interface is also arranged in the middle of the tank body, which can be used to connect a liquid level sensor device to monitor the inventory of the raw liquid to be processed in real time. When the liquid level is lower than the preset lower limit, the sensor outputs an alarm signal, and forms a chain triggering mechanism with the pump valve linkage logic of the whole system, so as to automatically terminate the subsequent liquid transfer and processing flow.

[0086] In the magnetic bead continuous separation system described in the present application, the waste liquid tank T3 serves as a receiving and temporary storage unit for supernatant in the magnetic separation process, and is communicated with the first liquid discharge port 13 of the magnetic bead separation device M1 through the fifth flow channel 105, for collecting the waste solution discharged in the separation operation without containing target magnetic beads. Such waste liquid mainly includes impurity components not captured by the magnetic field and non-target particles in the magnetic bead raw liquid.

[0087] The waste liquid tank T3 can adopt a vertical or horizontal double-elliptical head container in the structural design, and the specific structural form can be flexibly adjusted according to the installation space and flux demand. The tank body is provided with a liquid level meter interface, which can be externally connected to a liquid level sensor to realize dynamic monitoring of the liquid level in the tank. When the waste liquid level reaches the set upper limit value, an automatic control signal is output through the interlocking alarm system to trigger the pump body, valve or control device associated with the system to stop running, so as to avoid the risk of liquid overflow or downstream pipeline back pressure problem caused by overfilling of the tank body.

[0088] A first magnetic bead sensor AE1 is arranged on the fifth flow channel 105 to detect the concentration level of the target magnetic beads in the waste liquid flowing through the path in real time. The sensor is configured in an online continuous monitoring mode and can be linked with the data acquisition and control module to realize process evaluation of separation efficiency and feedback determination of magnetic bead loss. In different embodiments, the first magnetic bead sensor AE1 can be a spectrophotometer, a particle size analyzer, a clarity detector, a turbidity detector, etc. Although the detection principles of these instruments are different, the system indirectly measures the target magnetic bead concentration by processing the measurement data.

[0089] 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. The inlet of the product tank T2 is connected to the second outlet 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.

[0090] In different embodiments, the product tank T2 can have a single-layer wall structure or a double-layer jacket structure. The single-layer wall structure has the advantage of low cost, and the inner cavity of the double-layer jacket can be connected to a heating and refrigeration circulation system for precise temperature control of the inside of the tank. By arranging a temperature sensor in the tank and realizing closed-loop control with the circulation device, the magnetic bead liquid can always be in a suitable constant temperature state. To further improve the cleaning ability of the equipment, a spray cleaning interface is arranged on the top of the tank to perform internal flushing operation after the system operation ends, effectively removing the magnetic bead liquid that may be left on the tank wall, and avoiding batch cross contamination.

[0091] At the same time, the product tank T2 is provided with a liquid level meter interface on the outer wall, which is used to connect a liquid level monitoring sensor to realize real-time detection of the change in the volume of the tank. When the filling of the magnetic bead liquid reaches the preset upper limit, the system sends an alarm signal through the interlocking control logic and simultaneously controls the front fluid input path to stop running. A second magnetic bead sensor AE2 is arranged on the sixth flow channel 106 to monitor the concentration of the magnetic bead liquid entering the product tank T2. The real-time data measured can be used to judge 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, etc. The system indirectly measures the magnetic bead concentration by processing the measurement data.

[0092] In the magnetic bead continuous separation system described in the present application, the stirred tank R2 is used as an intermediate container for the magnetic bead washing process. The inlet of the stirred tank R2 is connected to the second outlet 14 of the magnetic bead separation device M1 through the seventh flow channel 107 to receive the magnetic bead liquid after preliminary purification. The outlet of the stirred tank R2 is connected to the first inlet 11 of the magnetic bead separation device M1 through the second flow channel 102 to realize the circulation and washing operation of the magnetic beads.

[0093] The stirred tank R2 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°, i.e., the angle of repose is greater than the natural inclination angle formed by the magnetic bead particles in the 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 the loss of magnetic beads during the washing process.

[0094] The stirred tank R2 is internally configured with multiple groups of stirring paddles driven by a motor. The stirring module can be automatically operated by a "one-key start" mode for uniformly dispersing the magnetic bead particles. In different embodiments, the stirred tank R2 can also be a common single-walled structure or a double-jacketed structure. The jacket is connected to the heat exchange medium output by the heating and refrigeration circulating machine, and a temperature sensor is inserted into the inside of the tank to realize real-time temperature monitoring and precise constant temperature control during stirring. The top of the stirred tank R2 is provided with a spray port to facilitate cleaning of the magnetic bead liquid adhered to the inner wall of the tank after batch replacement or long-term shutdown.

[0095] In addition, to real-time control the liquid level state in the tank, the upper part of the stirred tank R2 is equipped with a liquid level meter interface, and a chain alarm mechanism is set at two states of preset full liquid and preset empty liquid. When the liquid level is over limit or the liquid is empty, the control system can immediately respond and intervene in the start-stop instruction of the upstream liquid inlet or downstream liquid outlet process through the chain mechanism.

[0096] The washing tank T1 is used to store and supply the washing liquid required in the magnetic bead separation process. Its liquid outlets are respectively 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 mode, 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 realize continuous or multiple rounds of magnetic bead washing operation in the system. The washing liquid can be different according to the different magnetic bead raw materials, such as deionized water, ethanol, salt solution, and high molecular polymer.

[0097] The washing tank T1 is also preferably a double-elliptical-head container, which can be flexibly configured as a vertical or horizontal arrangement according to the process layout. The lower part of the tank body is provided with two independent discharge outlets, one of which is connected to a centrifugal pump interface for efficiently extracting the washing liquid before washing the magnetic beads, and the other is used to provide the magnetic bead separator with a pre-liquid filling function to ensure that the magnetic bead separator is pre-filled with sufficient liquid before the processed raw liquid is introduced, so as to optimize the distribution state and desorption path of the magnetic bead particles under the action of the magnetic field.

[0098] In the present 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 in the raw liquid tank R1 to the magnetic bead separation device M1, and is provided with a first valve KV-1 arranged 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.

[0099] The second flow channel 102 is used to circulate the magnetic bead washing liquid in the stirred tank R2 back to the magnetic bead separation device M1, and is provided with a second valve KV-2 arranged away from the stirred tank R2. The end of the second valve KV-2 away from the stirred tank R2 is connected to the inlet end of the fourth valve KV-4 through the first pump body P1.

[0100] The third flow channel 103 is used to deliver the washing liquid from the washing tank T1 to the first liquid inlet 11 of the magnetic bead separation device M1, and is provided with a third valve KV-3 arranged away from the washing tank T1. 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, so as to be combined with the first flow channel 101 and the second flow channel 102 near the magnetic bead separation device M1, and the raw liquid and the washing liquid can be switched in at this position.

[0101] 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, and is provided with a second pump body P2 and a fifth valve KV-5. The second pump body P2 and the fifth valve KV-5 cooperatively control the injection amount of the washing liquid and provide driving force.

[0102] The fifth flow channel 105 is used to discharge the waste liquid from the first liquid outlet 13 of the magnetic bead separation device M1 and guide it to the waste liquid tank T3. The fifth flow channel 105 is provided with a first magnetic bead sensor AE1 and a sixth valve KV-6. The sixth valve KV-6 is used to control the flow before and after the waste liquid is discharged, and cooperates with the sensor signal to judge the effective capture rate of the target magnetic beads, so as to avoid the target magnetic beads being discharged with the waste liquid.

[0103] The sixth flow channel 106 is used to connect the second liquid outlet 14 of the magnetic bead separation device M1 to the product tank T2, and is provided with a second magnetic bead sensor AE2 and an eighth valve KV-8. The eighth valve KV-8 is arranged near the magnetic bead separation device M1 to ensure that only the magnetic bead liquid meeting the concentration standard is introduced into the product tank T2.

[0104] The seventh flow channel 107 is used to connect the second liquid outlet 14 of the magnetic bead separation device M1 to the stirring tank R2, and the ninth valve KV-9 is arranged on the seventh flow channel 107. The end of the ninth valve KV-9 away from the stirring tank R2 is connected to the end of the eighth valve KV-8 away from the product tank T2, and the seventh valve KV-7 is connected to the second liquid outlet 14 of the magnetic bead separation device M1, thereby forming a split-flow control node of the product and circulating flow path.

[0105] The pump bodies arranged in the above flow channels include the first pump body P1 (for example, a peristaltic pump) arranged on the first flow channel 101, the second flow channel 102 and the third flow channel 103, and the second pump body P2 (for example, a centrifugal pump) arranged on the fourth flow channel 104, which is used to provide flow driving capacity required in different stages. It should be noted that the flow channels can be independently arranged or adaptively combined according to process requirements in different embodiments. For example, the first flow channel 101, the second flow channel 102 and the third flow channel 103 can share a combined pipeline near the magnetic bead separation device M1, so as to simplify the pipeline structure, reduce the liquid stagnation volume and improve the overall integration.

[0106] In the continuous automatic magnetic bead separation system described in the present application, the first pump body P1 is linked with the fourth valve KV-4 and the sixth valve KV-6. When the system stops running, the magnetic bead liquid can be discharged only from the second liquid outlet 14 of the magnetic bead separation device M1 by closing the valves, so as to avoid backflow. The second pump body P2 is used to provide a large flow liquid driving force when the pipeline is flushed, and 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 of different pipeline structures. In order to realize automatic control, the second pump body P2 and the first pump body P1 are mutually arranged in a start-stop interlocking manner. When the centrifugal pump runs, the first pump body P1 is in a stopped state, so as to avoid forming disturbance flow or pressure fluctuation in the pipeline system. In addition, the second pump body P2 is also linked with the fifth valve KV-5 and the seventh valve KV-7. When the second pump body P2 stops, only waste liquid (supernatant) is allowed to flow out from the first liquid outlet 13; and when the second pump body P2 starts, the magnetic bead liquid is flushed out from the second liquid outlet 14, so as to realize directional transfer of the magnetic beads. Further, the start-stop operation of the second pump body P2 also constitutes a logic interlocking with the demagnetization / magnetization behavior of the magnetic field generating device. When the magnetic bead release operation (demagnetization) is performed, the second pump body P2 is started; and when the magnetic separation process (magnetization) is resumed, the second pump body P2 stops running, and the system returns to the magnetic bead circulating washing process driven by the first pump body P1.

[0107] The system also has a plurality of detection instruments, such as a first magnetic bead sensor AE1 and a second magnetic bead sensor AE2, which include a spectrophotometer, a particle size analyzer, a clarity detector, and a turbidity detector, and are used to monitor the concentration of target substances in the magnetic bead liquid, the particle size of the magnetic beads, the clarity of the solution, and the turbidity of the washing liquid, so as to calculate the concentration of the magnetic beads according to the detection results and automatically adjust the working state of the pump body and the valve body. In addition, the first pump body P1 can be replaced by a metering pump in different embodiments, or can be operated in interlocking with an independent flow control circuit to adapt to different processing requirements. The operating parameters of all pump bodies in the system can be set according to different separation conditions, including the feeding rate of the magnetic bead liquid and the flushing flow rate of the washing liquid.

[0108] The embodiments of the present application also disclose a control method of a magnetic bead continuous separation system, referring to Figures 5-7 , comprising the following steps:

[0109] The device reset step: in response to a reset operation instruction, the components are controlled to enter an initial state; including sequentially controlling the first valve KV-1 to be closed, the second valve KV-2 to be closed, the third valve KV-3 to be closed, the fourth valve KV-4 to be closed, the fifth valve KV-5 to be closed, the sixth valve KV-6 to be closed, the seventh valve KV-7 to be closed, the eighth valve KV-8 to be closed, and the ninth valve KV-9 to be closed; 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; and controlling the magnetic bead separation device M1 to perform a demagnetization (demagnetization) operation.

[0110] The preset step: setting a first preset magnetic bead concentration, a second preset magnetic bead concentration, and a third preset magnetic bead concentration; setting a first preset lower limit height, a second preset upper limit height, and a second preset lower limit height; and setting the stirring speed and stirring time of the raw liquid tank R1 and the stirring kettle R2.

[0111] The magnetic bead separation device pre-liquid step: controlling the second valve KV-2 to be closed, the third valve KV-3 to be opened, the fourth valve KV-4 to be opened, and the sixth valve KV-6 to be opened, and controlling the magnetic bead separation device M1 to start the first pump body P1 after being magnetized; detecting whether the flow sensor detects flow, if not, continue to detect, if yes, go to the next step.

[0112] The single-batch raw liquid feeding step: controlling the third valve KV-3 to be closed, the first valve KV-1 to be opened, and detecting whether the concentration of the target magnetic beads detected by the first magnetic bead sensor AE1 reaches the first preset magnetic bead concentration, if not, continue to detect, if yes, go to the next step; wherein the target magnetic beads are particles that can be effectively captured by the magnetic field of the magnetic bead separation device M1.

[0113] Single batch magnetic bead pipetting step: control the first pump body P1 to stop, the fourth valve KV-4 to close, and the sixth valve KV-6 to close; control the fifth valve KV-5, the seventh valve KV-7, and the ninth valve KV-9 to open in a chain control manner, and control the magnetic bead separation device M1 to demagnetize; control the second pump body P2 to start, and control the stirring kettle R2 to start; acquire the liquid level detection signal of the liquid level sensor in the stirring kettle R2, and determine whether the liquid level of the stirring kettle R2 is higher than the second preset upper limit height; if not, continue to detect; if yes, proceed to the next step.

[0114] Single batch magnetic bead stirring and washing step: control the second pump body P2 to stop; 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 in a chain control manner; control the stirring kettle R2 to start and stir for a predetermined time.

[0115] 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 control the first pump body P1 to start; acquire the liquid level detection signal of the liquid level sensor in the stirring kettle R2, and determine whether the liquid level of the stirring kettle R2 is lower than the second preset lower limit height; if not, continue to detect; if yes, proceed to the next step.

[0116] Cleanliness detection step: stop the stirring kettle R2, and draw a two-dimensional data graph about time based on the concentration of the target magnetic beads detected by the first magnetic bead sensor AE1 in the waste liquid discharge step; determine whether the peak value 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.

[0117] Single batch magnetic bead liquid product pipetting step: control the first pump body P1 to stop, and 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 in a chain control manner; control the magnetic bead separation device M1 to demagnetize; control the second pump body P2 to start, and determine whether the concentration of the target magnetic beads detected by the second magnetic bead sensor AE2 is lower than the third preset magnetic bead concentration; if not, continue to detect; if yes, proceed to the next step.

[0118] Batch cycle step: control the fifth valve KV-5, the seventh valve KV-7, and the eighth valve KV-8 to close, and control the second pump body P2 to stop; acquire 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 magnetic bead separation device pre-charging step.

[0119] Pipe self-cleaning step: 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 height sensor in the stirring kettle R2 and judge whether it is higher than the second preset upper limit, until it is controlled to open the fourth valve KV-4, the sixth valve KV-6, and the first pump body P1 is started; based on the liquid level detection signal, and judge whether the liquid level height in the stirring kettle R2 is lower than the second preset lower limit height, if not, continue to detect, if yes, complete the pipe self-cleaning.

[0120] Device shutdown step: after detecting the production task end signal, perform the shutdown operation, 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, the ninth valve KV-9 to close; control the first pump body P1 to stop, the second pump body P2 to stop; control the raw liquid tank R1 to stop, the stirring kettle R2 to stop; control the magnetic bead separation device M1 to perform demagnetization operation, so as to complete the system shutdown and maintain the safe and static state of the device.

[0121] It should be noted that in different embodiments, the magnetic bead separation device pre-charging step of the above magnetic bead continuous separation system control method can be deleted, which plays an improving effect on the operation of the whole method but is not decisive, and correspondingly, the return step in the batch cycle step corresponds to the single batch raw liquid feeding step.

[0122] The application also discloses the following three different configurations of magnetic bead continuous separation system operation examples.

[0123] Example 1: vertical six-sided magnetic tile magnetic bead continuous separation system (deionized water washing)

[0124] Configuration characteristics

[0125] 1. Magnetic bead separation device: vertical structure, electric push rod lifting

[0126] 2. Magnetic attraction device: permanent magnet six-sided tile (magnetic field strength 1.0T)

[0127] 3. Washing liquid: deionized water

[0128] 4. Detection instrument: clarity detector (AE1 / AE2, unit: NTU (scattering turbidity))

[0129] 5. Magnetic bead parameters: particle size 1.5 μm, material Fe3O4

[0130] Key parameter setting

[0131]

[0132]

[0133] Flow time statistics:

[0134] 1. Single washing cycle time consumption: 60 min (including steps 4→9)

[0135] 2. Product purity: 98.9%

[0136] 3. Product recovery rate: 97.3%

[0137] Example 2: Horizontal electromagnetic magnetic bead continuous separation system (ethanol washing)

[0138] Configuration characteristics

[0139] 1. Magnetic bead separation device: horizontal structure, air cylinder lifting

[0140] 2. Magnetic attraction device: electromagnetic device (maximum magnetic field strength 30T)

[0141] 3. Washing liquid: ethanol (concentration 70%)

[0142] 4. Detection instrument: spectrophotometer (AE1 / AE2, unit OD600 (600nm wavelength optical density)) 5. Magnetic bead parameters: particle size 60μm, material carboxyl modified Fe3O4

[0143] Key parameter settings

[0144] Parameter name Set value AE1 high concentration value 0.8 OD600 AE1 low concentration value 0.1 OD600 AE2 low concentration value 0.05 OD600 LIT1 low liquid level value 0.2L LIT2 high liquid level value 100L LIT2 low liquid level value 0.2L R1 agitation speed 50 rpm R2 agitation speed 120 rpm R2 agitation duration 30 min P1 speed 80 mL / min

[0145] Flow time statistics

[0146] 1. Single washing cycle time consumption: 120 min (including steps 4→9)

[0147] 2. Product purity: 99.2%

[0148] 3. Product recovery rate: 97.6%

[0149] Example 3: Horizontal permanent magnet rhombic magnetic tile magnetic bead continuous separation system (high polymer polymer washing)

[0150] Configuration characteristics

[0151] 1. Magnetic bead separation device: horizontal structure, hydraulic transmission lifting

[0152] 2. Magnetic attraction device: permanent magnet rhombic tile (magnetic field strength 0.4T)

[0153] 3. Washing liquid: PEG-6000 (molecular mass of 6000) solution (concentration 5% w / v)

[0154] 4. Detection instrument: Turbidity detector (AE1 / AE2, unit FTU (scattering turbidity unit))

[0155] 5. Magnetic bead parameters: Particle size 0.8 pm, material SiO2 coated Fe3O4

[0156] Key parameter settings

[0157] Parameter name Set value AE1 high concentration value 100 FTU AE1 low concentration value 10 FTU AE2 low concentration value 2 FTU LIT1 low liquid level value 0.5L LIT2 high liquid level value 1000L LIT2 low liquid level value 0.5L R1 agitation speed 50 rpm R2 agitation speed 120 rpm R2 agitation duration 30 min P1 speed 80 mL / min

[0158] Flow time statistics:

[0159] 1. Single washing cycle time: 240 min (high viscosity washing liquid prolongs pipetting time) (including magnetic bead separation device pre-charging → cleanliness detection step)

[0160] 2. Product purity: 96.4%

[0161] 3. Product recovery rate: 98.5%

[0162] It can be seen that the continuous automatic magnetic bead separation system provided in the application has obvious advantages. The traditional magnetic bead separation process relying on manual operation has significant economic and efficiency disadvantages. First, in manual repetitive operation, as the processing scale expands, the required labor cost increases exponentially. For example, taking the processing of 1000 samples per day as an example, at least 10 operators are required to work in shifts, each working 8 hours a day, and at a rate of 50 yuan / hour, the daily labor cost is 5000 yuan. Using the automatic magnetic bead separation system, only one monitor is needed to complete the same work load, and the cost is reduced to about 200 yuan / day, which fundamentally saves labor costs. In addition, continuous manual operation also faces the problems of operation fatigue and psychological burden, and the error rate increases significantly from 0.1% to 5% after a long period of operation. The sample retesting, repeated reagent investment and management burden caused thereby further increase the monthly operating cost by an additional 10,000 yuan.

[0163] In terms of throughput, the processing capacity of manual operation is also difficult to meet the industrial sample processing demand, and usually each operator can process at most 200 samples per day, and the overall throughput is limited and difficult to expand. Industrial applications usually require continuous high-throughput output of thousands of samples per day, which is obviously beyond the capacity of manual operation. At the same time, in terms of purity and separation efficiency, the manual mode is prone to uneven distribution of magnetic bead particles due to large container size and uncontrollable fluid turbulence, which can cause significant fluctuations in target recovery rate, with a fluctuation amplitude of more than 30%, seriously affecting the consistency and quality stability of the final product.

[0164] To solve the above problems, the continuous automatic magnetic bead separation system provided by the application realizes the "raw material in-product out" full-process automatic processing path from raw liquid feeding to magnetic bead liquid product output by systematically integrating the magnetic bead separation device, the stirred tank, the automatic valve and the fluid control network, combining the online concentration monitoring and the liquid level control strategy, and constructing the multi-level automatic control logic. The core parameters such as flow, concentration, temperature and liquid level are controlled in real time through the logical linkage between the processing nodes, the interference of human operation intervention on the separation effect is effectively avoided, the processing efficiency and sample consistency are greatly improved, and the flux bottleneck and precision limitation of manual operation are fundamentally broken through.

[0165] Although the embodiments of the application have been shown and described above, it should be understood that the above embodiments are exemplary and should not be construed as limiting the application, and those of ordinary skill in the art can make changes, modifications, replacements and variations to the above embodiments within the scope of the application.

Claims

1. A magnetic bead continuous separation system, characterized by, The application relates to a magnetic bead separation device and a magnetic bead separation method. 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), the other end is provided with a first liquid outlet (13) and a second liquid outlet (14), the first liquid inlet (11) is away from the tube wall area capable of being magnetized and demagnetized relative to the second liquid inlet (12), and the first liquid outlet (13) is away from the tube wall area capable of being magnetized and demagnetized relative to the second liquid outlet (14); The raw liquid tank R1 is connected to the first liquid inlet (11) through a first flow channel (101); The waste liquid tank T3 is connected to the first liquid outlet (13) through a fifth flow channel (105), and the fifth flow channel (105) is provided with a first magnetic bead sensor AE1; The product tank T2 is connected to the second liquid outlet (14) through a sixth flow channel (106), and the sixth flow channel (106) is provided with a second magnetic bead sensor AE2; The stirring kettle R2 is connected to the second liquid outlet (14) through a seventh flow channel (107), and the liquid outlet of the stirring kettle R2 is connected to the first liquid inlet (11) through a second flow channel (102); The washing liquid tank T1 is connected to the first liquid inlet (11) through a third flow channel (103) and 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 of claim 1, wherein, The first valve KV-1 is arranged on the first flow channel (101), the second valve KV-2 is arranged on the second flow channel (102), the third valve KV-3 is arranged on the third flow channel (103), 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 kettle 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, the other end of the fourth valve KV-4 is connected to the first liquid inlet (11); the second pump body P2 and the fifth valve KV-5 are arranged on the fourth flow channel (104); the flow sensor and the sixth valve KV-6 are arranged on the fifth flow channel (105); the eighth valve KV-8 is arranged on the sixth flow channel (106), the ninth valve KV-9 is arranged on the seventh flow channel (107), and one end of the eighth valve KV-8 away from the product tank T2 and one end of the ninth valve KV-9 away from the waste liquid tank T3 are connected to the second liquid outlet (14) through the seventh valve KV-7.

3. The magnetic bead serial separation system according to claim 1, characterized in that, The liquid level sensor is correspondingly provided with a second preset upper limit and a second preset lower limit.

4. The magnetic bead serial separation system according to claim 1, characterized in that, The magnetic bead separation device M1 is used for separating a to-be-separated raw liquid containing target magnetic beads, the target magnetic beads are particles that can be effectively captured by an 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 serial separation system of claim 1, wherein, The magnetic bead separation device M1 comprises a magnetic bead separator (1), a magnetic field generating device (2), and a driving device (3) for driving the magnetic field generating device (2) to approach or move 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 outlet (13) and the second liquid outlet (14) are arranged at the other end of the magnetic bead separator (1).

6. A control method of a magnetic bead continuous separation system, for the magnetic bead continuous separation system according to any one of claims 2, comprising the following steps: Single-batch raw liquid feeding step: control the third valve KV-3 to be closed, the first valve KV-1 to be opened, and detect whether the concentration of the target magnetic beads detected by the first magnetic bead sensor AE1 reaches the first preset magnetic bead concentration, if not, continue to detect, if yes, proceed to the next step; wherein the target magnetic beads are particles that can be effectively captured by the magnetic field of the magnetic bead separation device M1. Single-batch magnetic bead pipetting step: control the first pump body P1 to stop, the fourth valve KV-4 to be closed, and the sixth valve KV-6 to be closed; control the fifth valve KV-5, the seventh valve KV-7, and the ninth valve KV-9 to be opened in a chain control manner, and control the magnetic bead separation device M1 to be demagnetized; control the second pump body P2 to be started in a chain control manner, and control the stirring kettle R2 to be started; acquire the liquid level detection signal of the liquid level sensor in the stirring kettle R2, and judge whether the liquid level of the stirring kettle R2 is higher than the second preset upper limit, if not, continue to detect, if yes, proceed to the next step. Single-batch magnetic bead stirring and washing step: control the second pump body P2 to stop; control the fifth valve KV-5, the seventh valve KV-7, and the ninth valve KV-9 to be closed in a chain control manner; start the stirring kettle R2 and stir for a predetermined time. Waste liquid discharging step: control the first valve KV-1 to be closed, the second valve KV-2 to be opened, the fourth valve KV-4 to be opened, and the sixth valve KV-6 to be opened; control the magnetic bead separation device M1 to be magnetized, and control the first pump body P1 to be started; acquire the liquid level detection signal of the liquid level sensor in the stirring kettle R2, and judge whether the liquid level of the stirring kettle R2 is lower than the second preset lower limit, if not, continue to detect, if yes, proceed to the next step. Cleanliness detection step: stop the stirring kettle R2, and draw a two-dimensional data graph about time based on the concentration of the target magnetic beads detected by the first magnetic bead sensor AE1 in the waste liquid discharging step; detect whether the peak value 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. Single batch magnetic bead liquid product pipetting step: control the first pump body P1 to stop, control the fourth valve KV-4 and the sixth valve KV-6 to close; interlock 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 the target magnetic bead detected by the second magnetic bead sensor AE2 is lower than the third preset magnetic bead concentration, if not, continue to detect, if yes, go to the next step; Batch cycle step: control the fifth valve KV-5, the seventh valve KV-7 and the eighth valve KV-8 to close, control the second pump body P2 to stop, obtain the liquid level height detection signal of the liquid level height sensor in the raw liquid tank R1, and judge whether the liquid level height 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 by, Further comprising the following steps: Pipeline self-cleaning step: control the raw liquid tank R1 to stop working, start the stirring kettle R2, control the fifth valve KV-5, the seventh valve KV-7 and the ninth valve KV-9 to open, start the second pump body P2; obtain the liquid level height detection signal of the liquid level height sensor in the stirring kettle R2, and judge whether the liquid level height in the stirring kettle R2 is higher than the second preset upper limit, until it is controlled to open the fourth valve KV-4 and the sixth valve KV-6, and the first pump body P1 starts; based on the liquid level height detection signal and judging whether the liquid level is lower than the second preset lower limit height, if not, continue to detect, if yes, complete the pipeline self-cleaning.

8. The control method of the magnetic bead continuous separation system according to claim 7, characterized by, Further comprising the following steps: Presetting step, setting the first setting value, the second setting value and the third setting value; set the first preset lower limit height, the second preset upper limit height and the second preset lower limit height; set 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, wherein, Before the single batch raw liquid feeding step, further comprising the following steps: Magnetic bead separator pre-liquid step: control the second valve KV-2 to close, the third valve KV-3 to open, the fourth valve KV-4 to open, the sixth valve KV-6 to open, control the magnetic bead separation device M1 to start the first pump body P1 after magnetizing; detect whether the flow sensor detects flow, if not, continue to detect, if yes, go to the next step.

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