Magnetic bead incubation method, device, equipment and medium

Abstract

The invention relates to the technical field of biomedical treatment, and discloses a magnetic bead incubation method which comprises the following steps: carrying out cell sorting and concentration on a cell sample in a centrifugal container to obtain concentrated cell sap; adding an incubation solution into the centrifugal container containing the concentrated cell sap, mixing, and performing magnetic bead incubation on the obtained mixed cell sap to obtain a target cell sap; and sub-packaging the target cell sap into a sub-packaging container. Separation of a large number of cell samples and concentration of the cell samples are achieved, non-specific binding of magnetic bead incubation is reduced by adding incubation liquid, the effect of magnetic bead incubation is improved, then the efficiency of magnetic bead incubation is improved, and meanwhile the pollution risk in the magnetic bead incubation process is greatly reduced.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to a method, device, equipment and medium for magnetic bead incubation. Background Art

[0002] At present, cell therapy is a key area of focus in the international medical frontier. During the process of cell therapy, magnetic bead incubation technology is one of the key steps.

[0003] In the prior art, immunomagnetic bead technology is currently widely used in immunodetection, cell separation, biomacromolecule passivation, biology and other aspects. Currently, the magnetic bead incubation method often manually binds target cells to magnetic bead-antibodies, and uses the complex of target cells and magnetic beads or the complex of target cells, antibodies and magnetic beads as the final product to achieve magnetic bead incubation. The target cells need to be manually sorted from cell samples, and there is an intermediate transfer process, resulting in problems such as contamination risk, poor incubation effect, low cell activity and low recovery rate. Therefore, how to provide a fully enclosed process from cell samples to magnetic bead incubation and an automated process with good incubation effect is an urgent problem for those skilled in the art in the current field. Summary of the Invention

[0004] Embodiments of the present invention provide a method, equipment and medium for magnetic bead incubation to solve the problems of contamination risk, poor incubation effect, low cell activity and low recovery rate existing in the magnetic bead incubation method in the prior art during the operation process.

[0005] A magnetic bead incubation method includes:

[0006] Performing cell sorting and concentration on a cell sample in a centrifuge container to obtain a concentrated cell solution;

[0007] Adding an incubation solution to the centrifuge container containing the concentrated cell solution, mixing, and then performing magnetic bead incubation on the obtained mixed cell solution to obtain a target cell solution;

[0008] Sub-packaging the target cell solution into a sub-packaging container.

[0009] A magnetic bead incubation device includes:

[0010] A sorting module for performing cell sorting and concentration on a cell sample in a centrifuge container to obtain a concentrated cell solution;

[0011] An incubation module for adding an incubation solution to the centrifuge container containing the concentrated cell solution, mixing, and then performing magnetic bead incubation on the obtained mixed cell solution to obtain a target cell solution;

[0012] A sub-packaging module for sub-packaging the target cell solution into a sub-packaging container.

[0013] A magnetic bead incubation device includes a centrifuge container and a controller connected to the centrifuge container, and the controller is used to execute the magnetic bead incubation method described above.

[0014] A computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the above-mentioned magnetic bead incubation method is implemented.

[0015] In the above-mentioned magnetic bead incubation method, device and medium, in the magnetic bead incubation method of the present invention, cell sorting and concentration are performed on the cell sample in the centrifuge container to obtain a concentrated cell solution; after adding an incubation solution to the centrifuge container containing the concentrated cell solution and mixing, magnetic bead incubation is performed on the obtained mixed cell solution to obtain a target cell solution; the target cell solution is dispensed into a dispensing container.

[0016] The magnetic bead incubation method of the present invention performs magnetic bead incubation operations in a fully enclosed device, thus avoiding the risk of contamination in manual operations and improving the safety during the magnetic bead incubation process. By performing sorting operations through this device, large-scale processing of cell samples is achieved, and the separation time of cell samples is reduced. By adding an incubation solution, the incubation effect is improved, and the recovery rate and cell activity of target cells in the final product are relatively high, improving the efficiency of magnetic bead incubation. Furthermore, the magnetic bead incubation method not only has strong compatibility with the instruments and consumables used, but also the instruments and consumables used can meet the requirements of different production lines and are easy to operate. Description of the Drawings

[0017] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the description of the embodiments of the present invention will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present invention, and those of ordinary skill in the art can also obtain other drawings without creative efforts based on these drawings.

[0018] Figure 1 is a flowchart of the magnetic bead incubation method in an embodiment of the present invention;

[0019] Figure 2 is a principle block diagram of the magnetic bead incubation device in an embodiment of the present invention;

[0020] Figure 3 is a schematic diagram of the magnetic bead incubation device in an embodiment of the present invention. Detailed Embodiments

[0021] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0022] In one embodiment, as Figure 1 shown, a method for incubating magnetic beads is provided, including the following steps:

[0023] S10: Perform cell sorting and concentration on the cell sample in the centrifuge container to obtain concentrated cell liquid.

[0024] It can be understood that the concentrated cell liquid refers to the high-concentration cell liquid obtained by centrifugal washing. The centrifuge container is a container for holding cell samples, including but not limited to centrifuge cups, centrifuge tanks, centrifuge bottles, etc. The cell sample is a sample including one or more of T cells, T cell subsets, and / or T cell progenitor cells.

[0025] Specifically, after the cell sample is input into the centrifuge container, the centrifuge container 10 is rotated and centrifuged at a preset speed so that various cells in the cell sample are separated and different cells are in different separation layers. By extracting the separated PBMC layer into the intermediate product liquid bag 16 or discharging other separation layers into the waste liquid bag 17, an intermediate product liquid is obtained. The centrifuge container 10 is washed with a cleaning solution to reduce the risk of contamination and cross-contamination. Then, the intermediate product liquid in the intermediate product liquid bag 16 is pumped back into the washed centrifuge container 10 by the peristaltic pump 7, and a washing and concentration operation is performed on the centrifuge container 10 to obtain concentrated cell liquid.

[0026] S20: After adding an incubation solution to the centrifuge container containing the concentrated cell liquid and mixing, perform magnetic bead incubation on the obtained mixed cell liquid to obtain a target cell liquid.

[0027] It can be understood that the target cell liquid is obtained by incubating target cells. The target cells include T cells and / or T cell subsets. The incubation solution refers to a magnetic bead incubation Buffer, and the magnetic bead incubation Buffer is a special solution used to promote the binding of magnetic beads to cells during the magnetic bead incubation process.

[0028] Specifically, after obtaining the concentrated cell solution, an incubation solution is added to the centrifuge container 10 containing the concentrated cell solution, and then the centrifuge container 10 is started to mix the concentrated cell solution and the incubation solution evenly. Then, magnetic bead incubation is performed on the obtained mixed cell solution, that is, a mixed solution of an antibody and magnetic beads is added to the centrifuge container 10. Among them, the magnetic beads can be micron-sized magnetic beads (magnetic beads with a particle size in the range of 1 to 5 microns) or nano-sized magnetic beads (magnetic beads with a particle size in the range of 5 to 100 nanometers). During the process of the pipeline transporting to the centrifuge container 10, the control valve 2 is opened, and the antibody and magnetic beads flow into the pipeline and then are input into the centrifuge container 10. The centrifuge container 10 can rotate back and forth at a specific speed so that the cells in the mixed cell solution bind to the antibody and magnetic beads, that is, the antibody and magnetic beads specifically bind to the antigen molecules on the cell surface in the concentrated cell solution, thereby obtaining the target cell solution.

[0029] S30: Dispense the target cell solution into a dispensing container.

[0030] Understandably, the dispensing container refers to a container for storing the target cell solution. For example, product bags of different specifications, etc.

[0031] Specifically, after performing magnetic bead incubation on the obtained mixed cell solution to obtain the target cell solution, at this time, the obtained target cell solution only contains T cells and / or T cell subsets; furthermore, the target cell solution can be dispensed into the dispensing container 15, for example, dispensed into multiple preparation bags. Further, directly dispensing the target cell solution into the dispensing container 15 may cause the cell activity in the target cell solution to decrease over time. Therefore, before dispensing the target cell solution into the dispensing container 15, a cryopreservation solution is input into the centrifuge container 10 containing the target cell solution to perform cryopreservation treatment on the target cell solution through the cryopreservation solution, so that the cryopreservation solution and the target cell solution can be mixed and then dispensed into the dispensing container 15 to maintain cell activity. Among them, the cryopreservation solution is resuspended with the target cells, and the centrifuge container is rotated left and right to mix the cryopreservation solution and the target cell solution, and then dispensed into the dispensing container 15.

[0032] Among them, sensors can be set on all pipeline paths in the present invention according to requirements for pipeline monitoring, and control valves can be set on all pipeline paths in the present invention according to requirements for pipeline opening and closing. For example, solenoid valves can be used for opening and closing the input / output of each pipeline.

[0033] The magnetic bead incubation method in the present invention performs the magnetic bead incubation operation in a fully enclosed device, thus avoiding the contamination risk in manual operation and improving the safety during the magnetic bead incubation process. By using this device to perform the sorting operation, large-scale processing of cell samples is achieved, and the separation time of cell samples is reduced. By adding the incubation solution, the incubation effect is enhanced, and the recovery rate and cell activity of target cells in the final product are relatively high, improving the efficiency of magnetic bead incubation. Furthermore, the magnetic bead incubation method not only has strong compatibility with the instruments and consumables used, but also the instruments and consumables used can meet the requirements of different production lines and are easy to operate.

[0034] In one embodiment, in step S10, that is, for the cell sample in the centrifuge container to perform cell sorting and concentration to obtain the concentrated cell solution, it includes:

[0035] S101, perform the product separation operation to separate the intermediate product solution from the cell sample in the centrifuge container.

[0036] It is understandable that the intermediate product solution refers to the PBMC solution or the T cell solution.

[0037] The PBMC solution is a cell solution containing peripheral blood mononuclear cells (PBMC). PBMC refers to the cells with a single nucleus in peripheral blood, including lymphocytes, monocytes, dendritic cells and other small amounts of cells. Lymphocytes include B cells and T cells.

[0038] Specifically, the cell sample is input from the sample liquid bag into the centrifuge container in the target device (the pipelines installed on this device are closed and sterile, and can make the centrifuge container in a constant rotating state) through the pipeline connected to the centrifuge container, that is, by starting the peristaltic pump 7, opening the control valves 4 and 13 of the pipeline, so as to pump the cell sample into the centrifuge container 10. Close the control valve 4 of this pipeline, open the control valve 5 on the pipeline connected to the sample separation liquid, add an appropriate sample separation liquid to the centrifuge container 10, and control the centrifuge container 10 to perform centrifugation at a preset speed. That is, by the density gradient centrifugation method, the cell sample will be stratified according to density and cell size, separating out the PBMC layer (mainly composed of mononuclear cells) and the supernatant layer (mainly composed of plasma and platelets). The intermediate product solution is extracted from the liquid outlet near or located in the PBMC layer. During the process, the speed can be continuously adjusted to fully extract the intermediate product solution in the centrifuge container 10, that is, open the control valve 6, and transfer the intermediate product solution to the intermediate product bag 16 through the peristaltic pump 7 via the connecting pipeline.

[0039] S102, clean the centrifuge container after the intermediate product solution has been separated.

[0040] Further, after separating the intermediate product liquid, the centrifuge container 10 is cleaned, that is, a cleaning liquid is introduced to clean the centrifuge container 10 and the pipeline. The flow direction of the cleaning liquid can be adjusted according to the design requirements of the pipeline. A control valve is provided in the pipeline (this control valve can be a solenoid valve to automatically control the opening and closing. Subsequently, when referring to the control valve, it can be a solenoid valve). By controlling the control valve, the opening and closing of different positions in the pipeline path are controlled, so that the cleaning liquid can clean each pipeline path, and the unnecessary components in the centrifuge container 10 are flushed into the waste liquid bag 17. Among them, the cleaning liquid can be a mixture of physiological saline and 0.5% HSA (human serum albumin).

[0041] S103, after the intermediate product liquid is transfused back into the centrifuge container, a washing operation is performed on the intermediate product liquid in the centrifuge container to obtain concentrated cell liquid.

[0042] Specifically, after the cell sample is input into the centrifuge container, the control valve 6 and the control valve 14 are opened. Then, the intermediate product liquid in the intermediate product bag 16 is refluxed into the centrifuge container 10 through the peristaltic pump 7. The peristaltic pump 7 can be reversed for transmission (for example: when the peristaltic pump draws, it rotates clockwise, and when it refluxes, it rotates counterclockwise). Then, the centrifuge container 10 is controlled to rotate centrifugally, so that the intermediate product liquid in the centrifuge container 10 is re-layered. That is, the opened control valve 1 is used to input at least one portion of the cleaning liquid into the centrifuge container 10, so that the components in the supernatant layer combine with the cleaning liquid (hydrophilic combination) to form a layer. Thus, the liquid containing the combined layer of the above components can be drawn from the liquid outlet into the waste liquid bag 17. By continuously adjusting the rotation speed of the centrifuge container 10, the liquid of the concentrated PBMC layer can be left, and this liquid is determined as the concentrated cell liquid. That is, in this embodiment, through the product separation operation, the separation of the intermediate product liquid is realized. By cleaning the container, the pollution risk is reduced. By performing the washing operation after transfusing the intermediate product liquid back into the centrifuge container, the concentration of the cell liquid is realized, and the acquisition of the concentrated cell liquid is realized.

[0043] In one embodiment, in step S101, that is, performing a product separation operation to separate the intermediate product liquid from the cell sample in the centrifuge container, the centrifuge container includes at least one liquid pushing plate that can rotate synchronously with the centrifuge container; the intermediate product liquid includes a first intermediate product liquid and a second intermediate product liquid; it includes:

[0044] S1011, after adding a first separation volume of sample separation liquid into the centrifuge container, the cell sample in the sample liquid bag is input into the centrifuge container.

[0045] S1012, the residual cell sample in the sample liquid bag is cleaned with the cleaning liquid, and the obtained cleaning mixture after cleaning is input into the centrifuge container.

[0046] Understandably, the centrifugal container 10 includes at least one liquid pushing plate that can rotate synchronously with the centrifugal container 10. The liquid pushing plate is arranged inside the centrifugal container 10. One end of the liquid pushing plate is close to the rotation axis, and the other end extends towards the inner side wall of the centrifugal container 10; there is a gap between one end of the liquid pushing plate close to the inner side wall of the centrifugal container 10 and the inner side wall; one end of the liquid pushing plate close to the bottom of the centrifugal container 10 is in contact with the bottom; when there are multiple liquid pushing plates, the multiple liquid pushing plates are circumferentially distributed relative to the rotation axis. The intermediate product liquid includes a first intermediate product liquid and a second intermediate product liquid. Among them, the intermediate product liquid may further include a third intermediate product liquid and a fourth intermediate product liquid, and the number of times of extracting the intermediate product liquid can be set according to the actual situation. The first intermediate product liquid refers to the intermediate product liquid separated for the first time. The second intermediate product liquid refers to the intermediate product liquid separated for the second time or the last time.

[0047] Specifically, install the consumables, input the consumable information, and perform the consumable self-check items, that is, detect the target device after connecting the consumables to complete the airtightness detection of the consumables and the sensor detection to ensure the aseptic sealing of the consumables. First, add a first separation volume (the first separation volume refers to a volume that can cover the entire bottom of the centrifugal container 10 and has a certain height, for example, 100 ml to 150 ml, etc.) of sample separation liquid into the centrifugal container 10, that is, start the centrifugal container 10 and rotate it at a preset speed (for example, 2100 rpm). Then, open the control valves between the sample separation liquid bag and the centrifugal container 10, that is, open the fifth control valve 5 and the fourteenth control valve 14 required for the connecting pipeline, and pump the first separation volume of sample separation liquid into the centrifugal container 10 through the peristaltic pump 7 to buffer the subsequent input cell samples. Then, close the fifth control valve 5 and the fourteenth control valve 14, and input the cell samples in the sample liquid bag (the input volume of the cell samples is set according to the specific situation, for example, 0 - 200 ml) into the centrifugal container 10, that is, keep the rotation speed of the centrifugal container 10, open the control valves between the sample liquid bag and the centrifugal container 10, that is, open the fourth control valve 4 and the thirteenth control valve 13 required for the connecting pipeline, and pump the cell samples into the centrifugal container 10 at a certain flow rate (this flow rate can be set according to the actual situation) through the peristaltic pump 7. In this way, the first separation volume of sample separation liquid helps the cell samples to cope with the changes in the internal and external environments, maintain the stability of the internal environment, and maintain the activity of the cells. Further, open the control valves between the cleaning liquid bag and the sample liquid bag, that is, open the first control valve 1, the fourth control valve 4, and the eighth control valve 8, backflush the cleaning liquid in the cleaning liquid bag into the sample liquid bag for cleaning, and pump the cleaned cleaning mixture into the centrifugal container 10 through the peristaltic pump 7 to ensure that all the cell samples are input into the centrifugal container 10. Among them, the entire input process is carried out under a constant temperature condition, for example, 4°C to 8°C, etc.

[0048] After adding a second separation volume of sample separation liquid to the centrifuge container containing the cell sample, control the rotation of the centrifuge container at a first separation rotation speed to perform a first sample separation operation on the cell sample, and push the cell sample to rotate synchronously along the same rotation direction as the centrifuge container through the liquid pushing plate.

[0049] After the first sample separation operation is performed for a first separation duration, extract a first volume of a first intermediate product liquid from the centrifuge container.

[0050] Understandably, the first separation rotation speed refers to the rotation speed for performing the first sample separation operation, which is set according to the actual situation. For example, it is 2500 rpm. The first separation duration refers to the time for performing the first sample separation operation, such as 200 s, etc.

[0051] Specifically, maintain the rotation speed of the centrifuge container 10, open the control valves between the sample separation liquid bag and the centrifuge container 10, that is, open the fifth control valve 5 and the fourteenth control valve 14 required for connecting the pipeline, and add a second separation volume of sample separation liquid to the centrifuge container 10 through the peristaltic pump 7. Then, close the fifth control valve 5 and the fourteenth control valve 14, and control the rotation of the centrifuge container 10 at a first separation rotation speed to perform a first sample separation operation on the cell sample, that is, the rotation speed can be increased to separate the target cells in the cell sample. Among them, the liquid pushing plate pushes the cell sample to rotate synchronously along the same rotation direction as the centrifuge container 10, thereby improving the efficiency and viability of the cells during the layering process of the cell sample. After the first sample separation operation is performed for a first separation duration (set this time according to the actual situation, such as 180 s), extract a first volume of a first intermediate product liquid from the centrifuge container 10, that is, the centrifuge container 10 rotates at the first separation rotation speed for the first separation duration, and then, reduce the rotation speed, open the fourteenth control valve 14 and the sixth control valve 6, and extract a certain volume (this volume can be set in advance, such as 100 ml) of the first intermediate product liquid to the intermediate product liquid bag 16 through the peristaltic pump 7 at a fixed flow rate.

[0052] After adding a third separation volume of sample separation liquid to the centrifuge container, control the rotation of the centrifuge container at a second separation rotation speed to perform a second sample separation operation on the cell sample, and push the cell sample to rotate synchronously along the same rotation direction as the centrifuge container through the liquid pushing plate.

[0053] After the second sample separation operation is performed for a second separation duration, extract a second volume of a second intermediate product liquid from the fully separated centrifuge container.

[0054] Understandably, the first separation speed refers to the speed at which the first sample separation operation is performed, which is set according to the actual situation. For example, it is 1500 rpm. The first separation duration refers to the time for performing the first sample separation operation, such as 80 s, etc.

[0055] Specifically, after the first intermediate product liquid is output, the sixth control valve 6 is closed, the rotation speed is maintained, the fifth control valve 5 is opened, and the sample separation liquid of the third separation volume is added to the centrifuge container 10 again through the peristaltic pump 7, and then the fifth control valve 5 and the fourteenth control valve 14 are closed. Then, the centrifuge container 10 is rotated by the second separation speed (this speed can be the same as or different from the first separation speed) to perform the second sample separation operation on the cell sample, that is, to fully separate the target cells in the cell sample, and the cell sample is pushed by the liquid pushing plate to rotate synchronously along the same rotation direction as the centrifuge container 10, so that the cell sample and the sample separation liquid are fully contacted. Further, after the second separation duration of the second sample separation operation is performed, the second separation speed is maintained, the fourteenth control valve 14 and the sixth control valve 6 are opened, and a certain volume (which can be determined according to the volume of the cell sample and the volume extracted for the first time. For example, if the volume of the cell sample is 150 ml and the volume extracted for the first time is 100 ml, then the volume extracted for the second time is 60 ml; if the volume of the cell sample is 120 ml and the volume extracted for the first time is 80 ml, then the volume extracted for the second time is 50 ml) of the second intermediate product liquid is extracted into the intermediate product liquid bag 16 through the peristaltic pump 7 at a fixed flow rate, that is, the second volume of the second intermediate product liquid is extracted from the fully separated centrifuge container 10. That is, in this embodiment, by adding the sample separation liquid of the first separation volume to the centrifuge container, the buffering of the cell sample is realized, avoiding cell damage and reducing cell activity. By adding the sample separation liquid multiple times, the full separation of the cells in the cell sample is realized, and the preparation of the intermediate product liquid is realized.

[0056] In one embodiment, in the step S103, that is, performing a washing operation on the intermediate product liquid in the centrifuge container to obtain a concentrated cell liquid, includes:

[0057] S1031, adding a cleaning liquid to the centrifuge container containing the intermediate product liquid, and controlling the rotation of the centrifuge container by the first cleaning speed to perform a primary cleaning on the intermediate product liquid to obtain a cleaned cell liquid, and recording the number of cleaning times as one.

[0058] S1032. Perform the repeated cleaning operation for a preset number of times. The repeated cleaning operation includes: adding the cleaning solution to the cell solution after cleaning again, and controlling the centrifuge container to rotate at a second cleaning rotation speed to repeatedly clean the cell solution after cleaning, obtaining the cell solution after repeated cleaning, and incrementing the cleaning count by one. The second cleaning rotation speed is greater than the first cleaning rotation speed, and the second cleaning rotation speed corresponding to each repeated cleaning operation is greater than the second cleaning rotation speed corresponding to the previous repeated cleaning operation.

[0059] S1033. After the cleaning count reaches the preset cleaning count, confirm that the repeated cleaning operation is completed.

[0060] S1034. Perform a washing operation on the cell solution obtained after the last repeated cleaning in the centrifuge container to obtain a concentrated cell solution.

[0061] Understandably, the preset number of times refers to the number of cleaning times set in advance, for example, 3 times, etc. The second cleaning rotation speed is greater than the first cleaning rotation speed, and the second cleaning rotation speed corresponding to each repeated cleaning operation is greater than the second cleaning rotation speed corresponding to the previous repeated cleaning operation. The first cleaning rotation speed refers to the rotation speed of the centrifuge container when cleaning the intermediate product solution for the first time, for example, 400 rpm. The second cleaning rotation speed refers to the rotation speed of the centrifuge container when cleaning the intermediate product solution again, for example, 600 rpm. The first cleaning time refers to the duration of the centrifuge container when cleaning the intermediate product solution for the first time. The second cleaning time refers to the duration of the centrifuge container when cleaning the intermediate product solution again.

[0062] Specifically, after the separated intermediate product liquid is transfused back from the intermediate product liquid bag 16 to the centrifuge container 10, that is, after the centrifuge container 10 and the pipeline are cleaned and the waste liquid is rinsed into the waste liquid bag 17, the sixth control valve 6 and the thirteenth control valve 13 are opened, and the separated intermediate product liquid is transfused back into the centrifuge container 10 through the peristaltic pump 7, and then the sixth control valve 6 is closed. A cleaning liquid is added to the centrifuge container 10 containing the intermediate product liquid, that is, the first control valve 1 is opened, and a certain volume (set according to the actual situation, for example, 30 ml) of the cleaning liquid is added to the centrifuge container 10 through the peristaltic pump 7. After the input is completed, the first control valve 1 and the thirteenth control valve 13 are closed. Then, the centrifuge container 10 is started and the rotation speed is increased to the first cleaning rotation speed, and the centrifuge container 10 is controlled to rotate at the first cleaning rotation speed (set according to the actual situation, for example, 300 rpm) to perform the primary cleaning of the intermediate product liquid, that is, to dissolve the metabolites and impurities in the intermediate product liquid in the cleaning liquid. After maintaining the first cleaning time, the liquid is layered, and the waste liquid discharging operation is performed on the centrifuge container 10, that is, the fourteenth control valve 14 and the eleventh control valve 11 are opened, and the waste liquid is discharged into the waste liquid bag 17 through the peristaltic pump 7, so as to obtain the cleaned cell liquid retained in the centrifuge container 10, and the cleaning times are recorded as one. Then, it is judged whether the cleaning times reach the preset times. When the cleaning times reach the preset times, the cleaning operation ends and the next process is entered. When the cleaning times do not reach the preset times, the repeated cleaning operation of the preset times is performed. The repeated cleaning operation includes: adding the cleaning liquid to the cleaned cell liquid again, and controlling the centrifuge container 10 to rotate at the second cleaning rotation speed to perform repeated cleaning on the cleaned cell liquid to obtain the repeatedly cleaned cell liquid, and increasing the cleaning times by one, that is, raising the first cleaning rotation speed to the second cleaning rotation speed, opening the first control valve 1 and the thirteenth control valve 13, and adding a certain volume of the cleaning liquid to the centrifuge container 10 at a fixed flow rate through the peristaltic pump 7, and maintaining the second cleaning rotation speed. After centrifuging for the second cleaning time, the waste liquid discharging operation is performed on the centrifuge container 10, that is, the fourteenth control valve 14 and the eleventh control valve 11 are opened, and the waste liquid is discharged into the waste liquid bag 17 through the peristaltic pump 7, and concentrated to the minimum volume (this volume is set in advance), that is, a preset volume of cell liquid is retained in the centrifuge container 10. After the cleaning times do not reach the preset cleaning times, the rotation speed is continuously increased and the volume of the cleaning liquid is reduced for cleaning. After the cleaning times reach the preset cleaning times, it is confirmed that the repeated cleaning operation is completed. Further, a washing operation is performed on the cell liquid obtained after the last repeated cleaning in the centrifuge container 10, that is, the metabolites or impurities on the cells in the cell liquid are washed, and the fourteenth control valve 14 and the twelfth control valve 12 are opened to perform the waste liquid discharging operation, and a preset volume of cell liquid is retained in the centrifuge container 10, so as to obtain the concentrated cell liquid.

[0063] That is, in this embodiment, by cleaning the intermediate product liquid a preset number of times, the pollutants in the intermediate product liquid are removed, and the purity of the cells in the intermediate product liquid is improved. By performing a washing operation on the cell liquid obtained after the last repeated cleaning in the centrifuge container, the pollutants and impurities on the cell surface are removed, and at the same time, the cells are concentrated, and the cell recovery rate is improved.

[0064] In one embodiment, in step S1034, that is, performing a washing operation on the cell liquid obtained after the last repeated cleaning in the centrifuge container to obtain a concentrated cell liquid, includes:

[0065] S1031, adding a cleaning liquid with a first washing volume to the cell liquid after the last repeated cleaning in the centrifuge container, and controlling the centrifuge container to rotate at a first washing rotation speed to wash the immune cells in the cell liquid to obtain a washed cell liquid.

[0066] S1032, adding a buffer solution with a second washing volume to the washed cell liquid, and controlling the centrifuge container to rotate at a second washing rotation speed to obtain the concentrated cell liquid after the washing is completed.

[0067] It can be understood that the washed cell liquid refers to the cell liquid obtained by washing the cell liquid. The first washing volume refers to the volume of the cleaning liquid set in advance, for example, 120 ml, etc. The second washing volume refers to the volume of the incubation solution or buffer solution set in advance, for example, 200 ml, etc.

[0068] Specifically, after obtaining the cell fluid after the last repeated washing, increase the centrifugation speed, for example, increase it to the first washing speed (e.g., 1900 rpm). Then, open the first control valve 1 and the thirteenth control valve 13, add a first washing volume of cleaning fluid to the cell fluid after the last repeated washing in the centrifuge container 10, and precisely control the volume through the sensor in the pipeline. Close the first control valve 1 and the thirteenth control valve 13, and control the centrifuge container 10 to perform centrifugal rotation at the first washing speed to wash the immune cells in the cell fluid, that is, maintain the first washing speed. After performing the first washing time (e.g., 300 s), perform the waste liquid discharging operation, that is, open the fourteenth control valve 14 and the twelfth control valve 12, discharge the waste liquid in the centrifuge container 10 into the waste liquid bag 17, and leave a preset reserved volume (e.g., 30 ml, etc.) of cell fluid in the centrifuge container 10. After the waste liquid discharging ends, close the fourteenth control valve 14 and the twelfth control valve 12, thereby obtaining the washed cell fluid. Further, switch the liquid bag at the interface corresponding to the first control valve 1 to the buffer solution bag (or the magnetic bead incubation Buffer solution bag), open the first control valve 1 and the thirteenth control valve 13, add a second washing volume of buffer solution to the washed cell fluid through the peristaltic pump 7, and control the centrifuge container 10 to perform centrifugal rotation at the second washing speed, that is, first rotate the centrifuge container 10 at a low speed for a certain time to mix the washed cell fluid and the buffer solution. Then, increase the centrifugation speed, maintain a certain rotation speed for a certain time, and then decrease the centrifugation speed. Open the fourteenth control valve 14 and the eleventh control valve 11, perform the waste liquid discharging operation, and after the waste liquid discharging ends, close the fourteenth control valve 14 and the eleventh control valve 11, and leave a preset volume of cell fluid in the centrifuge container 10, thereby obtaining the concentrated cell fluid after washing is completed.

[0069] That is, in this embodiment, by respectively performing the cleaning and washing operations on the cell fluid after the last repeated washing, the removal of contaminants and impurities on the cell surface is achieved, and at the same time, the concentration of the target cells is realized, the purity and recovery rate of the target cells are improved, and then the efficiency and effect of subsequent magnetic bead incubation are improved by using the magnetic bead incubation Buffer.

[0070] In one embodiment, in the step S20, that is, performing magnetic bead incubation on the obtained mixed cell fluid to obtain the target cell fluid, includes:

[0071] S201, add magnetic beads and antibodies to the centrifuge container so that the magnetic beads and antibodies specifically bind to the cells in the mixed cell fluid to obtain the intermediate cell fluid.

[0072] S202. Add a buffer solution to the centrifuge container containing the intermediate cell solution to centrifugally remove the unbound magnetic beads and antibodies in the intermediate cell solution. After centrifugation, draw out the supernatant and drain the waste liquid to a preset retention volume to obtain the target cell solution.

[0073] Understandably, the intermediate cell solution refers to the cell solution after the magnetic beads and antibodies are coupled to the target cells. The preset retention volume refers to the volume retained in the centrifuge container after draining the waste liquid, which is set in advance.

[0074] Specifically, after obtaining the concentrated cell solution, lower the temperature of the surrounding environment of the centrifuge container 10 to keep the environment around the centrifuge container 10 at 4°C to 8°C, which can reduce non-specific binding and make the effect of magnetic bead incubation better. Then, add the incubation solution to the centrifuge container 10 containing the concentrated cell solution and mix them, that is, add the incubation solution to start the centrifuge container 10 to mix evenly to promote the binding of the magnetic beads and antibodies to the cells. Then, add magnetic beads and antibodies to the centrifuge container 10 so that the magnetic beads and antibodies specifically bind to the cells in the mixed cell solution, that is, the antibody is directly or indirectly coupled to the surface of the magnetic bead to form an affinity magnetic bead complex, so that the magnetic bead complex binds to the target cells in the mixed cell solution. That is, rotate the centrifuge container 10 to evenly mix the mixed cell solution with the magnetic bead complex, and the liquid pushing plate pushes the mixed cell solution so that the magnetic bead complex binds to the target cells. That is, the magnetic bead complex is directly or indirectly coupled with an antibody on the surface, specifically recognizes the antigen on the surface of the target cell, and binds to the antigen on the surface of the target cell, so that the magnetic bead complex is directly connected to the target cell to form a magnetic label, thereby obtaining the intermediate cell solution. Among them, the magnetic beads include nano magnetic beads and micro magnetic beads. The nano magnetic beads can be separated or not separated during subsequent use, and the micro magnetic beads must be separated during subsequent use.

[0075] Further, a certain volume (set according to actual conditions, for example, 200 ml) of buffer solution is added to the centrifuge container 10 containing the intermediate cell fluid. Then, a preset volume of cleaning solution is added to clean the intermediate cell fluid after specific binding. That is, the cleaning operation is repeatedly performed by adding the cleaning solution for a preset number of cleaning times to first remove impurities and metabolites in the cell fluid, and then centrifugally remove the unbound magnetic beads and antibodies in the intermediate cell fluid. After a period of time, the liquid is layered, and then the supernatant is drawn out, and the fourteenth control valve 14 and the twelfth control valve 12 are opened to perform the waste liquid discharging operation until the waste liquid is discharged to the preset remaining volume, thereby obtaining the target cell fluid. That is, in this embodiment, by adding magnetic beads and antibodies to the centrifuge container 10, specific binding of the magnetic beads and antibodies to the cells in the mixed cell fluid is achieved, and magnetic bead incubation of the target cells is realized. By adding buffer solution and cleaning solution to the centrifuge container 10 containing the intermediate cell fluid, cleaning of impurities is achieved, and removal of unbound magnetic beads and antibodies is realized, thereby realizing the preparation of the target cell fluid, reducing the pollution risk, and improving the purity and recovery rate.

[0076] In another embodiment, in step S20, that is, performing magnetic bead incubation on the obtained mixed cell fluid to obtain the target cell fluid, it further includes:

[0077] S203, adding magnetic beads and antibodies to the centrifuge container containing the mixed cell fluid, and performing primary incubation on the mixed cell fluid to obtain an immunocyte fluid.

[0078] S204, adding magnetic beads and antibodies to the centrifuge container containing the immunocyte fluid again, and performing secondary incubation on the immunocyte fluid to obtain the target cell fluid.

[0079] It can be understood that the immunocyte fluid refers to the one obtained by the first binding of magnetic beads and antibodies to target cells.

[0080] Specifically, magnetic beads and antibodies are added to the centrifuge container 10 containing the mixed cell solution. That is, first, the interface corresponding to the second control valve 2 is connected to the liquid bag containing the magnetic beads and antibodies, and then the second control valve 2, the ninth control valve 9, and the thirteenth control valve 13 are opened. The magnetic beads and antibodies are input into the centrifuge container 10 containing the mixed cell solution through a peristaltic pump. Among them, the interface corresponding to the fifth control valve 5 is connected to the liquid bag containing the buffer solution. The fifth control valve 5 and the eighth control valve 8 are opened to wash the liquid bag containing the magnetic beads and antibodies, and the washed liquid is input into the centrifuge container 10 through a pipeline. Then, the mixed cell solution is incubated once. That is, the antibody is directly or indirectly coupled to the surface of the magnetic beads to form an affinity magnetic bead complex. Then, the centrifuge container 10 is rotated to uniformly mix the mixed cell solution with the magnetic bead complex, so that the magnetic bead complex binds to the target cells, thereby obtaining an immunocyte solution. Further, magnetic beads and antibodies are added again to the centrifuge container 10 containing the immunocyte solution, and the immunocyte solution is incubated twice. That is, the second control valve 2, the ninth control valve 9, and the thirteenth control valve 13 are opened, magnetic beads and antibodies are added again, and then the rotation speed of the centrifuge container 10 is increased to uniformly mix the magnetic bead complex in the immunocyte solution, so as to fully bind the magnetic beads and antibodies to the target cells, thereby obtaining a target cell solution. Among them, the processes of the two incubations are the same, and the volume of the mixed solution of the added magnetic beads and antibodies and the rotation speed of the centrifuge container can be the same or different. That is, in this embodiment, by incubating the mixed cell solution multiple times, the preparation of the target cell solution is realized, and the effect of magnetic bead incubation and the recovery rate of cells are improved.

[0081] In one embodiment, the magnetic bead incubation method, that is, after adding an incubation solution to the centrifuge container containing the concentrated cell solution and mixing, before performing magnetic bead incubation on the obtained mixed cell solution, further includes:

[0082] S401, input buffer solution into the concentrated cell solution and mix, and control the concentrated cell solution in the centrifuge container to perform centrifugal rotation at a first preset rotation speed. After a first centrifugation time, sample a first sampling volume of the mixed cell solution.

[0083] S402, perform cell image acquisition on the sampled mixed cell solution through an image acquisition device to obtain a cell image.

[0084] S403, perform cell counting and recognition on the cell image through a cell recognition device to determine the counting result in the cell image.

[0085] Understandably, the first sampling volume refers to the volume of the extracted mixed cell fluid set in advance. The mixed cell fluid is obtained by mixing a buffer solution into the concentrated cell fluid. The counting result refers to the number of cells in the cell image. The cell image is obtained by photographing the sampled mixed cell fluid. The cell density refers to the number of cells per unit volume or unit area. The first preset rotation speed refers to the rotation speed for controlling the centrifuge container to mix the concentrated cell fluid and the buffer solution. The first sampling volume refers to the volume of the sampled mixed cell fluid, for example, 1 ml, etc.

[0086] Specifically, after obtaining the concentrated cell fluid, add a buffer solution to the centrifuge container, that is, open the first control valve 1 and the fourteenth control valve 14 of the pipeline, and start the peristaltic pump 7 to rotate counterclockwise to pump the buffer solution into the centrifuge container 10 at a fixed flow rate, and control the concentrated cell fluid and the buffer solution in the centrifuge container 10 to rotate centrifugally at the first preset rotation speed so that the concentrated cell fluid and the buffer solution are mixed evenly. After the first centrifugation time (for example, 200 s), sample the mixed cell fluid with the first sampling volume, that is, extract the mixed cell fluid with the first sampling volume. Then, perform cell image acquisition on the sampled mixed cell fluid through an image acquisition device, that is, photograph the preset detection device through the image acquisition device to obtain a cell image. Further, input the cell image into the cell recognition model, and extract the cell shape features in the cell image through the cell recognition model. The cell shape features are the features presented by the edge of the cell and the shape enclosed by the edge. Then, the cell recognition model performs cell recognition on the cell image according to the extracted cell shape features to obtain a cell region with cell shape features. The cell region can be the region of a single cell or the region enclosed by cell stacking or adjacency. Count the number of the recognized cell regions, that is, output the corresponding number of the regions of cell stacking or adjacency to obtain the counting result in the cell image. In this way, the cell recognition model can more accurately recognize the number of cells and improve the reliability of cell number recognition.

[0087] S404, obtain the image parameters corresponding to the cell image, and determine the cell density based on the counting result and the image parameters.

[0088] Understandably, the image parameters are the parameters required for density conversion of the cell number, such as the size of the cell image, etc. The cell density refers to the number of cells per unit volume.

[0089] Specifically, after determining the counting result in the cell image, the image parameters corresponding to the cell image are obtained. Based on the counting result and the image parameters, the cell density is calculated. When the image parameters are the image area and the image depth of field, by obtaining the set image area and image depth of field, the image volume of the collected image is calculated, and the counting result is divided by the image volume to obtain the cell density. That is, in this embodiment, by calculating the density of the mixed cell solution, the determination of the cell density is realized, and further the judgment of whether the number of cells meets the standard is realized, ensuring the number of target cells during magnetic bead incubation.

[0090] It should be understood that the magnitudes of the sequence numbers of the steps in the above embodiments do not mean the order of execution. The order of execution of each process should be determined according to its function and internal logic, and should not constitute any limitation to the implementation process of the embodiments of the present invention.

[0091] In one embodiment, a magnetic bead incubation device is provided, and this magnetic bead incubation device corresponds one-to-one to the magnetic bead incubation method in the above embodiment. As Figure 2 shown, this magnetic bead incubation device includes a sorting module 10, an incubation module 20, and a dispensing module 30. The detailed description of each functional module is as follows:

[0092] The sorting module 10 is used to perform cell sorting and concentration on the cell sample in the centrifuge container to obtain a concentrated cell solution;

[0093] The incubation module 20 is used to add an incubation solution to the centrifuge container containing the concentrated cell solution and then perform magnetic bead incubation on the obtained mixed cell solution to obtain a target cell solution;

[0094] The dispensing module 30 is used to dispense the target cell solution into a dispensing container.

[0095] Preferably, the sorting module 10 includes:

[0096] A separation unit for performing a product separation operation to separate an intermediate product solution from the cell sample;

[0097] A cleaning unit for cleaning the centrifuge container after the intermediate product solution has been separated;

[0098] A washing unit for performing a washing operation on the intermediate product solution in the centrifuge container after the intermediate product solution is returned to the centrifuge container to obtain a concentrated cell solution.

[0099] Preferably, the incubation module 20 includes:

[0100] A specific binding unit for adding magnetic beads and antibodies to the centrifuge container so that the magnetic beads and antibodies specifically bind to the cells in the mixed cell solution to obtain an intermediate cell solution;

[0101] A centrifugal removal unit for adding a buffer solution to the centrifuge container containing the intermediate cell solution to centrifugally remove the unbound magnetic beads and antibodies in the intermediate cell solution, and after centrifugation, extracting the supernatant and discharging the waste liquid to a preset retention volume to obtain the target cell solution.

[0102] Preferably, the incubation module 20 further includes:

[0103] A primary incubation unit for adding magnetic beads and antibodies to the centrifuge container containing the mixed cell solution and performing a primary incubation on the mixed cell solution to obtain an immunized cell solution;

[0104] A secondary incubation unit for adding magnetic beads and antibodies again to the centrifuge container containing the immunized cell solution and performing a secondary incubation on the immunized cell solution to obtain the target cell solution.

[0105] Preferably, the centrifuge container includes at least one liquid pushing plate that can rotate synchronously with the centrifuge container; the intermediate product solution includes a first intermediate product solution and a second intermediate product solution; the separation unit includes:

[0106] A sample input unit for adding a first separation volume of sample separation liquid to the centrifuge container and then inputting the cell sample in the sample liquid bag into the centrifuge container;

[0107] A residual cleaning unit for cleaning the residual cell sample in the sample liquid bag with a cleaning solution and inputting the obtained cleaning mixture into the centrifuge container;

[0108] A first sample separation unit for adding a second separation volume of sample separation liquid to the centrifuge container containing the cell sample, controlling the rotation of the centrifuge container at a first separation speed to perform a first sample separation operation on the cell sample, and pushing the cell sample to rotate synchronously along the same rotation direction as the centrifuge container through the liquid pushing plate;

[0109] A first separation unit for extracting a first volume of the first intermediate product solution from the centrifuge container after the first sample separation operation is performed for a first separation duration;

[0110] The second sample separation unit is configured to, after adding a third separation volume of sample separation liquid into the centrifuge container, control the rotation of the centrifuge container at a second separation speed to perform a second sample separation operation on the cell sample, and push the cell sample to rotate synchronously along the same rotation direction as the centrifuge container through the liquid pushing plate;

[0111] The second separation unit is configured to, after the second sample separation operation is performed for a second separation duration, extract a second volume of second intermediate product liquid from the sufficiently separated centrifuge container.

[0112] Preferably, the washing unit includes:

[0113] The primary cleaning unit is configured to add cleaning liquid into the centrifuge container containing the intermediate product liquid, control the rotation of the centrifuge container at a first cleaning speed to perform primary cleaning on the intermediate product liquid, obtain the cleaned cell liquid, and record the number of cleaning times as one;

[0114] The repeated cleaning unit is configured to perform a preset number of repeated cleaning operations. The repeated cleaning operation includes: adding cleaning liquid into the cleaned cell liquid again, and controlling the rotation of the centrifuge container at a second cleaning speed to perform repeated cleaning on the cleaned cell liquid, obtain the repeatedly cleaned cell liquid, and increment the number of cleaning times by one; the second cleaning speed is greater than the first cleaning speed, and the second cleaning speed corresponding to each repeated cleaning operation is greater than the second cleaning speed corresponding to the previous repeated cleaning operation;

[0115] The confirmation completion unit is configured to confirm the completion of the repeated cleaning operation after the number of cleaning times reaches the preset number of cleaning times;

[0116] The washing operation unit is configured to perform a washing operation on the cell liquid obtained after the last repeated cleaning in the centrifuge container to obtain concentrated cell liquid.

[0117] Preferably, the washing operation unit includes:

[0118] The cell washing sub-unit is configured to add a first washing volume of cleaning liquid into the cell liquid obtained after the last repeated cleaning in the centrifuge container, control the rotation of the centrifuge container at a first washing speed to wash the immune cells in the cell liquid, and obtain the washed cell liquid;

[0119] The completion washing sub-unit is configured to add a second washing volume of buffer solution into the washed cell liquid, and control the rotation of the centrifuge container at a second washing speed to obtain the concentrated cell liquid after completion of washing.

[0120] In one embodiment, as Figure 3As shown, a magnetic bead incubation device includes a centrifuge container and a controller connected to the centrifuge container. The controller is configured to execute the magnetic bead incubation method described above.

[0121] Specific limitations regarding the magnetic bead incubation device, the controller, and their respective units and modules can be referred to the limitations of the magnetic bead incubation method in the foregoing text and will not be elaborated herein. Each module in the above controller can be implemented in whole or in part by software, hardware, and their combination. Understandably, the controller includes a processor, a memory, a network interface, and a database connected through a device bus. Each module of the controller can be embedded in or independent of the processor in the form of hardware, or stored in the memory in the form of software for the processor to call and execute the operations corresponding to the above respective modules. Among them, the processor is used to provide computing and control capabilities. The memory includes a non-volatile storage medium and an internal memory. The non-volatile storage medium stores an operating device, a computer program, and a database. The internal memory provides an environment for the operation of the operating device and the computer program in the non-volatile storage medium. The database is used to store the data used in the magnetic bead incubation method in the above embodiments. The network interface is used to communicate with an external terminal through a network connection. The computer program, when executed by the processor, implements a magnetic bead incubation method.

[0122] In one embodiment, a computer-readable storage medium is provided. The computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the above magnetic bead incubation method is implemented.

[0123] Those of ordinary skill in the art can understand that all or part of the processes in the methods of the above embodiments can be completed by instructing relevant hardware through a computer program. The computer program can be stored in a non-volatile computer-readable storage medium. When the computer program is executed, it can include the processes of the embodiments of the above methods. Among them, any reference to a memory, storage, database, or other medium used in the embodiments provided in the present application can include non-volatile and / or volatile memories. Non-volatile memories can include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM), or flash memory. Volatile memories can include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM is available in various forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDR SDRAM), enhanced SDRAM (ESDRAM), synchronous link DRAM (SLDRAM), Rambus direct RAM (RDRAM), direct memory bus dynamic RAM (DRDRAM), and Rambus dynamic RAM (RDRAM), etc.

[0124] Those skilled in the art can clearly understand that for the convenience and brevity of description, only the above division of each functional unit and module is used as an example. In actual applications, the above functions can be allocated to different functional units and modules as needed, that is, the internal structure of the device can be divided into different functional units or modules to complete all or part of the functions described above.

[0125] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention, and should all be included in the protection scope of the present invention.

Claims

1. A method for incubating magnetic beads, characterized in that, Comprising: Performing cell sorting and concentration on a cell sample in a centrifuge container to obtain a concentrated cell solution; Adding an incubation solution to the centrifuge container containing the concentrated cell solution, mixing, and then performing magnetic bead incubation on the resulting mixed cell solution to obtain a target cell solution; Dispensing the target cell solution into a dispensing container.

2. The method for incubating magnetic beads according to claim 1, characterized in that, The performing cell sorting and concentration on a cell sample in a centrifuge container to obtain a concentrated cell solution includes: Performing a product separation operation to separate an intermediate product solution from the cell sample; Cleaning the centrifuge container after the intermediate product solution has been separated; After returning the intermediate product solution to the centrifuge container, performing a washing operation on the intermediate product solution in the centrifuge container to obtain a concentrated cell solution.

3. The method for incubating magnetic beads according to claim 1, characterized in that, The performing magnetic bead incubation on the resulting mixed cell solution to obtain a target cell solution includes: Adding magnetic beads and an antibody to the centrifuge container so that the magnetic beads and the antibody specifically bind to the cells in the mixed cell solution to obtain an intermediate cell solution; Adding a buffer solution to the centrifuge container containing the intermediate cell solution to centrifuge and remove the unbound magnetic beads and antibody in the intermediate cell solution, and after centrifugation, withdrawing the supernatant and discharging the waste liquid to a preset retention volume to obtain the target cell solution.

4. The method for incubating magnetic beads according to claim 1, characterized in that, The performing magnetic bead incubation on the resulting mixed cell solution to obtain a target cell solution further includes: Adding magnetic beads and an antibody to the centrifuge container containing the mixed cell solution, performing a first incubation on the mixed cell solution to obtain an immunized cell solution; Adding magnetic beads and an antibody again to the centrifuge container containing the immunized cell solution, performing a second incubation on the immunized cell solution to obtain the target cell solution.

5. The method for incubating magnetic beads according to claim 2, characterized in that, The centrifuge container includes at least one liquid pushing plate that can rotate synchronously with the centrifuge container; the intermediate product solution includes a first intermediate product solution and a second intermediate product solution; The performing a product separation operation to separate an intermediate product solution from a cell sample in a centrifuge container includes: After adding a first separation volume of a sample separation solution to the centrifuge container, inputting the cell sample in the sample liquid bag into the centrifuge container; Cleaning the residual cell sample in the sample liquid bag with a cleaning solution and inputting the resulting cleaning mixed solution into the centrifuge container; After adding a second separation volume of the sample separation solution to the centrifuge container containing the cell sample, controlling the rotation of the centrifuge container at a first separation speed to perform a first sample separation operation on the cell sample, and pushing the cell sample to rotate synchronously along the same rotation direction as the centrifuge container by the liquid pushing plate; After the first sample separation operation has been performed for a first separation duration, extracting a first volume of the first intermediate product solution from the centrifuge container; After adding a third separation volume of the sample separation solution to the centrifuge container, controlling the rotation of the centrifuge container at a second separation speed to perform a second sample separation operation on the cell sample, and pushing the cell sample to rotate synchronously along the same rotation direction as the centrifuge container by the liquid pushing plate; After the second sample separation operation is performed for a second separation duration, a second volume of a second intermediate product liquid is extracted from the centrifugation container after sufficient separation.

6. The method for incubating magnetic beads according to claim 2, characterized in that, Performing a washing operation on the intermediate product liquid in the centrifugation container to obtain a concentrated cell liquid, including: Adding a cleaning liquid to the centrifugation container containing the intermediate product liquid, controlling the centrifugation container to rotate at a first cleaning rotation speed to perform a primary cleaning on the intermediate product liquid, obtaining a cleaned cell liquid, and recording the number of cleaning times as one; Performing a preset number of repeated cleaning operations, the repeated cleaning operation including: adding the cleaning liquid to the cleaned cell liquid again, and controlling the centrifugation container to rotate at a second cleaning rotation speed to perform repeated cleaning on the cleaned cell liquid, obtaining a repeatedly cleaned cell liquid, and incrementing the number of cleaning times by one; the second cleaning rotation speed is greater than the first cleaning rotation speed, and the second cleaning rotation speed corresponding to each repeated cleaning operation is greater than the second cleaning rotation speed corresponding to the previous repeated cleaning operation; After the number of cleaning times reaches a preset number of cleaning times, confirm that the repeated cleaning operation is completed; Performing a washing operation on the cell liquid obtained after the last repeated cleaning in the centrifugation container to obtain a concentrated cell liquid.

7. The method for incubating magnetic beads according to claim 6, characterized in that, Performing a washing operation on the cell liquid obtained after the last repeated cleaning in the centrifugation container to obtain a concentrated cell liquid, including: Adding a cleaning liquid with a first washing volume to the cell liquid obtained after the last repeated cleaning in the centrifugation container, controlling the centrifugation container to rotate at a first washing rotation speed to wash the immune cells in the cell liquid, obtaining a washed cell liquid; Adding a buffer solution with a second washing volume to the washed cell liquid, controlling the centrifugation container to rotate at a second washing rotation speed to obtain the concentrated cell liquid after completion of washing.

8. A magnetic bead incubation device, characterized in that, Including: A sorting module for performing cell sorting and concentration on a cell sample in a centrifugation container to obtain a concentrated cell liquid; An incubation module for adding an incubation liquid to the centrifugation container containing the concentrated cell liquid, and after mixing, performing magnetic bead incubation on the obtained mixed cell liquid to obtain a target cell liquid; A dispensing module for dispensing the target cell liquid into a dispensing container.

9. A magnetic bead incubation device, characterized in that, Including a centrifugation container and a controller connected to the centrifugation container, the controller being configured to execute the magnetic bead incubation method according to any one of claims 1 to 7.

10. A computer-readable storage medium storing a computer program, characterized in that, The computer program, when executed by a processor, implements the magnetic bead incubation method according to any one of claims 1 to 7.

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