Protein purification device and method

Abstract

Disclosed in the present application are a protein purification device and method. The protein purification device comprises a rotary disc, a liquid adding module and a mixing module; a plurality of positioning seats are provided on one side of the rotary disc, the positioning seats can rotate together with the rotary disc, and the positioning seats are used for placing at least one container; the liquid adding module comprises at least one liquid outlet, the liquid outlet being used for adding a reagent into the container; and the mixing module comprises a magnetic rod sleeve and a magnetic rod, the magnetic rod being used for extending into the magnetic rod sleeve. The mixing module and the liquid adding module are both independent modules, and cooperate with the rotary disc having a plurality of stations, such that a mixing operation and a liquid adding operation can be automatically performed, thereby saving the total time for protein purification and improving the protein purification efficiency.

Description

Protein purification apparatus and methods

[0001] Cross-reference information

[0002] This application claims priority to Chinese patent application No. 202411538243.0, filed on October 30, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This application relates to the field of protein purification technology, specifically to a protein purification apparatus and method. Background Technology

[0004] A protein purification instrument is a laboratory instrument used to separate and purify proteins. Through various protein separation techniques, it isolates proteins from complex biological samples to obtain high-purity target proteins. Traditional protein purification steps include cell disruption, clarification, precipitation, chromatography, concentration, and storage. These steps typically require significant time and experimental expertise and demand highly skilled operators.

[0005] In recent years, novel technologies such as magnetic bead purification have been gradually introduced to improve protein purification efficiency and ease of operation. The semi-automatic purification system is an automated device based on magnetic bead purification. Because cell debris in cells and fermentation broth does not affect the binding of magnetic beads to the target protein and antibody, no additional centrifugation or filtration is required before adding the magnetic beads to the sample, thus significantly saving preliminary preparation work. After sample incubation is complete, the centrifuge tubes are transferred to the instrument for washing, elution, and regeneration.

[0006] However, the elution process requires manual sample collection and centrifuge tube replacement, which remains inconvenient and inefficient. Furthermore, commercially available protein purification instruments require manual liquid addition or the use of pre-packaged reagent kits (provided by the manufacturer), resulting in high operating costs. Users who use their own reagents require additional time to add the reagents to the containers one by one. Summary of the Invention

[0007] This application provides a protein purification apparatus and method to improve protein purification efficiency.

[0008] In a first aspect, this application provides a protein purification apparatus, including a turntable, a liquid addition module, and a mixing module. The turntable has multiple positioning seats on one axial side, spaced circumferentially along the turntable. The positioning seats are rotatable with the turntable and are used to hold at least one container. The liquid addition module and the positioning seats are located on the same axial side of the turntable; the liquid addition module includes at least one liquid outlet for adding reagents to the container. The mixing module and the liquid addition module are located on the same axial side of the turntable, spaced circumferentially along the turntable; the mixing module includes a magnetic rod sleeve and a magnetic rod, the magnetic rod extending into the magnetic rod sleeve.

[0009] In one specific implementation scheme, a plurality of the positioning seats are equally spaced along the circumference of the turntable; along the circumference of the turntable, the spacing angle between the mixing module and the liquid addition module is the same as the spacing angle between adjacent positioning seats.

[0010] In one specific implementation, the mixing module further includes a first mounting bracket and a second mounting bracket, which are respectively movable along the axial direction of the turntable. The projections of the first mounting bracket and the second mounting bracket at least partially overlap in the axial direction of the turntable. The magnetic rod is disposed on the side of the first mounting bracket away from the second mounting bracket, and the magnetic rod is disposed on the side of the second mounting bracket facing the first mounting bracket.

[0011] In one specific implementation, there are multiple magnetic rod sleeves, which are arranged in at least one row along the tangent of the turntable, with each row of magnetic rod sleeves including at least one magnetic rod sleeve; and there are multiple magnetic rods, which are arranged in at least one row along the tangent of the turntable, with each row of magnetic rods including at least one magnetic rod.

[0012] In one specific implementation, the number of magnetic rods is the same as the number of magnetic rod sleeves, and one magnetic rod is used to extend into one magnetic rod sleeve along the axial direction of the turntable.

[0013] In one specific implementation, along the axial direction of the turntable, a first end of the magnetic rod sleeve is provided with a insertion hole, the size of which is smaller than the size of the magnetic rod sleeve, and the projection of the magnetic rod is located within the insertion hole; the magnetic rod sleeve and the magnetic rod are respectively capable of moving along the axial direction of the turntable, the second end of the magnetic rod sleeve is used to extend into the container, and the magnetic rod is used to extend from the first end of the magnetic rod sleeve into the insertion hole to extend into the magnetic rod sleeve.

[0014] In one specific implementation, the turntable is provided with a mounting through hole, which is coaxially arranged with the turntable; the mixing module further includes a column extending along the axial direction of the turntable, the projection of the column on the axial direction of the turntable being located within the mounting through hole, and the turntable being rotatable relative to the column; a first driving part and a second driving part are provided inside the column, the first mounting bracket is connected to the first driving part, and the first driving part is used to drive the first mounting bracket to move along the axial direction of the turntable; the second mounting bracket is connected to the second driving part, and the second driving part is used to drive the second mounting bracket to move along the axial direction of the turntable.

[0015] In one specific implementation scheme, a mounting base is provided in the mounting through hole, and the turntable is rotatable relative to the mounting base; the column and the liquid addition module are located on the same side of the turntable in the axial direction, and one end of the column is fixedly connected to the mounting base.

[0016] In one specific implementation, the positioning seat is provided with a positioning frame, the positioning frame is provided with at least one positioning hole, the axis of the positioning hole is parallel to the axis of the turntable, and the positioning hole is used to accommodate the container.

[0017] In one specific implementation, the turntable is provided with multiple positioning protrusions, which form multiple positioning protrusion groups. The number of positioning protrusion groups is the same as the number of positioning seats. Each positioning protrusion group includes multiple positioning protrusions, and the multiple positioning protrusions included in each positioning protrusion group are arranged around a positioning seat.

[0018] In one specific implementation, the liquid filling module further includes a liquid filling needle for connection to a liquid filling pipeline. The liquid filling needle is capable of moving along the axial and radial directions of the turntable, and the liquid outlet is disposed on the liquid filling needle.

[0019] In one specific implementation, the liquid dispensing module further includes a third drive unit, a fourth drive unit, and a liquid dispensing needle holder. The third drive unit is located on one side of the turntable in the radial direction. The fourth drive unit and the positioning seat are located on the same side of the turntable in the axial direction. The fourth drive unit is connected to the third drive unit, and the third drive unit can drive the fourth drive unit to move along the axial direction of the turntable. The liquid dispensing needle holder is connected to the fourth drive unit, and the fourth drive unit can drive the liquid dispensing needle holder to move radially along the turntable. The liquid dispensing needle is disposed on the liquid dispensing needle holder.

[0020] In one specific implementation, the liquid injection needle holder is provided with a plurality of limiting holes, which are arranged in at least one row along the tangent of the turntable; and a liquid injection needle is disposed in one of the limiting holes.

[0021] In one specific implementation, the liquid dispensing module further includes a needle washing tank, which is disposed on the third drive unit. The needle washing tank is located on the side of the liquid dispensing needle holder facing the turntable. The side of the needle washing tank facing the liquid dispensing needle holder has an opening, through which the liquid dispensing needle is inserted into the needle washing tank.

[0022] Secondly, this application provides a method for purifying proteins using the aforementioned protein purification apparatus, comprising the following steps:

[0023] 1) Place the positioning seat containing the container at one station of the turntable. The container contains a sample to be purified and the sample contains magnetic beads. The remaining stations of the turntable contain the positioning seat containing an empty container.

[0024] 2) Control the magnetic rod sleeve of the mixing module to mix the sample in the container, and control the liquid addition module to add reagents to the empty container adjacent to the container in the rotation direction of the turntable;

[0025] 3) Control the magnetic rod of the mixing module to magnetically attract the sample that has been mixed in the container;

[0026] 4) Control the turntable to rotate one station, and control the magnetic rod of the mixing module to release the magnetically attracted sample into the container where the reagent has been added;

[0027] 5) Repeat steps 2) to 4) until the entire setup process is complete.

[0028] In one specific implementation, the reagent added to the container by the liquid addition module is a washing solution, an elution solution, a magnetic bead regeneration buffer, a washing buffer, or a storage buffer.

[0029] In one specific implementation, before replacing the reagent added to the container, the liquid dispensing module controls the liquid outlet to move to the needle washing tank, rinses the liquid outlet with the replaced reagent, or simultaneously rinses the liquid dispensing pipeline connected to the liquid outlet.

[0030] Compared with the prior art, the beneficial effects of this application are as follows:

[0031] The protein purification device provided in this application employs a dual-module design (liquid addition module and mixing module) and a turntable with multiple stations. The mixing and liquid addition modules are independent and can automatically perform mixing and liquid addition operations, thereby saving total protein purification time and improving purification efficiency. Furthermore, the required reagents can be added to containers at different stations via the liquid addition module to achieve different purposes such as washing, elution, or regeneration, thus increasing the automation level of the entire protein purification process. Attached Figure Description

[0032] Figure 1 shows a perspective view of the protein purification apparatus provided in this application in a certain state;

[0033] Figure 2 shows a perspective view of the protein purification apparatus provided in this application in another state;

[0034] Figure 3 shows a top view of the protein purification apparatus provided in this application;

[0035] Figure 4 shows a schematic diagram of other layout configurations of the workstation of the protein purification apparatus provided in this application;

[0036] Figure 5 shows a schematic diagram of other layout configurations of the workstation of the protein purification apparatus provided in this application;

[0037] Figure 6 shows a schematic diagram of other layout configurations of the workstation of the protein purification apparatus provided in this application;

[0038] Figure 7 shows a schematic diagram of a positioning frame of the protein purification apparatus provided in this application;

[0039] Figure 8 shows another structural schematic diagram of the positioning frame of the protein purification device provided in this application;

[0040] Figure 9 shows a partial structural schematic diagram of the protein purification apparatus provided in this application;

[0041] Figure 10 shows a schematic diagram of the mixing module of the protein purification apparatus provided in this application;

[0042] Figure 11 shows a schematic diagram of the liquid addition module of the protein purification apparatus provided in this application;

[0043] Figure 12 shows a schematic diagram of the liquid addition module of the protein purification apparatus provided in this application.

[0044] Reference numerals: 1-Turntable; 2-Mixing module; 3-Liquid addition module; 4-Positioning frame; 11-Positioning seat; 12-Positioning protrusion; 21-Second mounting frame; 211-Magnetic rod; 22-First mounting frame; 221-Magnetic rod sleeve; 23-Mounting base; 24-Column; 31-Liquid addition needle; 32-Third drive unit; 33-Needle washing tank; 34-Fourth drive unit; 35-Liquid addition needle holder; 41-First positioning frame; 42-Second positioning frame. Detailed Implementation

[0045] To make the objectives, technical solutions, and advantages of this application clearer, the embodiments of this application will be described in further detail below with reference to the accompanying drawings. However, the exemplary embodiments can be implemented in many forms and should not be construed as limited to the embodiments set forth herein. The same reference numerals in the figures denote the same or similar structures, and therefore repeated descriptions of them will be omitted. The terms expressing position and direction described in the embodiments of this application are illustrative based on the accompanying drawings, but changes can be made as needed, and all such changes are included within the scope of protection of this application. The accompanying drawings of the embodiments of this application are only for illustrating relative positional relationships and do not represent actual scale.

[0046] Specific details are set forth in the following description to aid in understanding this application; however, embodiments of this application can be implemented in various ways other than those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the embodiments of this application. Therefore, this application is not limited to the specific embodiments disclosed below.

[0047] The protein purification apparatus provided in this application embodiment can utilize magnetic bead purification to purify specific target proteins.

[0048] Figure 1 shows a perspective view of the protein purification apparatus provided in this application in one state, Figure 2 shows a perspective view of the protein purification apparatus provided in this application in another state, and Figure 3 shows a top view of the protein purification apparatus provided in this application. As shown in Figures 1 to 3, the protein purification apparatus provided in this application embodiment may include a turntable 1, a liquid addition module 3, and a mixing module 2. The turntable 1 can rotate around its own axis under the drive of a motor. Multiple positioning seats 11 are provided on one side of the turntable 1 along its own axis, and the multiple positioning seats 11 are spaced apart circumferentially along the turntable 1. The positioning seats 11 can rotate together with the turntable 1, and are used to place at least one container. Exemplarily, the number of positioning seats 11 is six, forming six workstations, which can be labeled as workstations S, A, B, C, D, and E respectively. The container can be a centrifuge tube.

[0049] The liquid addition module 3 and the positioning seat 11 are located on the same side of the turntable 1 in the axial direction. The liquid addition module 3 includes at least one liquid outlet for adding reagents into the container. The mixing module 2 and the liquid addition module 3 are located on the same side of the turntable 1 in the axial direction and are spaced apart circumferentially along the turntable 1.

[0050] In specific implementation, the mixing module 2 includes a magnetic rod sleeve 221 and a magnetic rod 211. Along the axial direction of the turntable 1, the first end of the magnetic rod sleeve 221 has an insertion hole, the size of which is smaller than the size of the magnetic rod sleeve 221 (it can be understood that the magnetic rod sleeve 221 is a tubular structure with a closed bottom). The projection of the magnetic rod 211 is located within the insertion hole. The magnetic rod sleeve 221 and the magnetic rod 211 can move along the axial direction of the turntable 1. The second end of the magnetic rod sleeve 221 is used to extend into the container, and the magnetic rod 211 is used to extend into the insertion hole from the first end of the magnetic rod sleeve 221, thus allowing the magnetic rod 211 to extend into the magnetic rod sleeve 221. The magnetic rod 211 has a magnetic attraction function. Specifically, the magnetic rod 211 can be a structure in which a magnet is encased in a metal shell. The magnet can be located at the bottom of the metal shell, meaning the bottom of the magnetic rod 211 has a relatively obvious magnetic attraction function. There can be multiple magnets; when the number of magnets is large, the overall magnetic attraction force of the magnetic rod 211 is greater.

[0051] The reagents in the container can be stirred by the reciprocating motion of the magnetic rod sleeve 221 along the axis of the turntable 1. When the magnetic rod 211 is inserted into the magnetic rod sleeve 221, the magnetic beads contained in the reagent can be adsorbed onto the outer surface of the magnetic rod sleeve 221 under the action of magnetic attraction. When the magnetic rod 211 is withdrawn from the magnetic rod sleeve 221, the magnetic beads are released from the magnetic rod sleeve 221. In this way, the adsorption and release of magnetic beads in the reagent can be achieved by the relative motion of the magnetic rod 211 and the magnetic rod sleeve 221. Thus, different reagents in the container can be used to achieve the purpose of washing and elution.

[0052] The protein purification apparatus provided in this application adopts a design that combines a dual-module (liquid addition module 3 and a mixing module 2) with a turntable 1 having multiple stations. Both the mixing module 2 and the liquid addition module 3 are independent modules, capable of automatically performing mixing and liquid addition operations. This saves the total time for protein purification and improves purification efficiency. Furthermore, the liquid addition module 3 can add the required reagents to containers at different stations to achieve different purposes such as washing, elution, or regeneration, thereby increasing the automation level of the entire protein and antibody purification process. Figure 1 illustrates the state where the mixing module 2 is mixing and the liquid addition module 3 is adding liquid; Figure 2 illustrates the state where the mixing module 2 is not mixing and the liquid addition module 3 is not adding liquid.

[0053] As one possible implementation, multiple positioning seats 11 can be equally spaced along the circumference of the turntable 1. For example, six positioning seats 11 form six workstations (S, A, B, C, D, and E workstations), with adjacent workstations spaced 60° apart. Along the circumference of the turntable 1, the interval angle between the mixing module 2 and the liquid addition module 3 can be the same as the interval angle between adjacent positioning seats 11. That is, the mixing module 2 corresponds to one of the six workstations, and the liquid addition module 3 corresponds to the other of the six workstations. This allows for simultaneous liquid addition and mixing, eliminating the need to separate the mixing and liquid addition operations, further improving process efficiency, and saving module space.

[0054] Figures 4, 5, and 6 illustrate other layout configurations of the workstations in the protein purification apparatus provided in this application. From the perspective of Figures 4, 5, and 6, the positioning seat 11 is obscured by the positioning frame 4. Although the positioning frame 4 is directly shown in the figures, the intended representation is the layout of the positioning seat 11, i.e., the layout of the workstations. As shown in Figure 4, two positioning seats 11 form two workstations. In this case, the mixing module 2 corresponds to one of the workstations, and the liquid addition module 3 corresponds to the other workstation. As shown in Figure 5, three positioning seats 11 form three workstations. In this case, the mixing module 2 corresponds to one of the three workstations, and the liquid addition module 3 corresponds to the other workstation. As shown in Figure 6, four positioning seats 11 form four workstations. In this case, the mixing module 2 corresponds to one of the four workstations, and the liquid addition module 3 corresponds to the other workstation. It is understood that the protein purification apparatus provided in the embodiments of this application may also have other numbers of workstations, such as nine or twelve workstations.

[0055] In practical implementation, the positioning seat 11 can be equipped with a positioning frame 4, which has at least one positioning hole. The axis of the positioning hole is parallel to the axis of the turntable 1, and the positioning hole can accommodate containers such as centrifuge tubes. In actual applications, the positioning frame 4 can accommodate containers of different volumes, such as 50mL and 100mL containers.

[0056] Figure 7 shows a schematic diagram of one structure of the positioning frame of the protein purification apparatus provided in this application, and Figure 8 shows another schematic diagram of the positioning frame of the protein purification apparatus provided in this application. Generally, when the container volume is large, the radial dimension of the container is also large. Adaptively, as shown in Figure 7, a first positioning frame 41 with a larger radial dimension of the positioning hole can be used. When the container volume is large, the total volume of the magnetic beads in the reagent added to the container can be large. In this case, a magnetic rod 211 with a larger magnetic attraction force can be used, that is, a magnetic rod 211 with a larger radial dimension can be used. Correspondingly, a magnetic rod sleeve 221 with a larger radial dimension can be used. Conversely, when the container volume is small, as shown in Figure 8, a second positioning frame 42 with a smaller radial dimension of the positioning hole can be used. When the container volume is small, the total volume of the magnetic beads in the reagent added to the container can be small. In this case, a magnetic rod 211 with a smaller magnetic attraction force can be used, that is, a magnetic rod 211 with a smaller radial dimension can be used. Correspondingly, a magnetic rod sleeve 221 with a smaller radial dimension can be used.

[0057] Figure 9 shows a partial structural schematic diagram of the protein purification apparatus provided in this application. As shown in Figure 9, the turntable 1 can be provided with multiple positioning protrusions 12, which form multiple positioning protrusion groups. The number of positioning protrusion groups is the same as the number of positioning seats 11. A positioning protrusion group includes multiple positioning protrusions 12, and the multiple positioning protrusions 12 included in a positioning protrusion group are arranged around a positioning seat 11. Thus, the positioning frame 4 on each positioning seat 11 can be individually limited by a positioning protrusion group, which can prevent the positioning frame 4 on each positioning seat 11 from shaking or displacing during the rotation of the turntable 1.

[0058] Figure 10 shows a schematic diagram of the mixing module of the protein purification apparatus provided in this application. As shown in Figure 10, the mixing module 2 may further include a first mounting frame 22 and a second mounting frame 21. The first mounting frame 22 and the second mounting frame 21 are respectively movable along the axial direction of the turntable 1. In the axial direction of the turntable 1, the projections of the first mounting frame 22 and the second mounting frame 21 at least partially overlap. A magnetic rod sleeve 221 is disposed on the side of the first mounting frame 22 away from the second mounting frame 21, and a magnetic rod 211 is disposed on the side of the second mounting frame 21 facing the first mounting frame 22. The relative movement of the first mounting frame 22 and the second mounting frame 21 allows the magnetic rod 211 to extend into or retract from the magnetic rod sleeve 221.

[0059] In specific implementation, there can be multiple magnetic rod sleeves 221, arranged in at least one row along the tangent of the turntable 1, with each row including at least one magnetic rod sleeve 221. Magnetic rod sleeves 221 belonging to a row can be connected as a whole, allowing for the installation and removal of the entire row of magnetic rod sleeves 221. Correspondingly, there can also be multiple magnetic rods 211, arranged in at least one row along the tangent of the turntable 1, with each row including at least one magnetic rod 211.

[0060] In practice, the number of magnetic rods 211 is the same as the number of magnetic rod sleeves 221. Along the axial direction of the turntable 1, one magnetic rod 211 can be inserted into or withdrawn from one magnetic rod sleeve 221. This allows for simultaneous mixing of reagents in multiple containers. For example, there are twelve magnetic rod sleeves 221 arranged in three rows, with four sleeves per row; the number and arrangement of the magnetic rods 211 are the same as those of the magnetic rod sleeves 221.

[0061] In actual setup, the turntable 1 can be provided with a mounting through hole, which is coaxially arranged with the turntable 1. The mixing module 2 can also include a column 24, which extends along the axial direction of the turntable 1. The projection of the column 24 on the axial direction of the turntable 1 is located within the mounting through hole, allowing the turntable 1 to rotate relative to the column 24. A first driving unit and a second driving unit are provided inside the column 24. A first mounting bracket 22 is connected to the first driving unit, which can drive the first mounting bracket 22 to move along the axial direction of the turntable 1. A second mounting bracket 21 is connected to the second driving unit, which can drive the second mounting bracket 21 to move along the axial direction of the turntable 1. By driving the first mounting bracket 22 and the second mounting bracket 21 to move respectively through the first driving unit and the second driving unit, the magnetic rod 211 can be inserted into the magnetic rod sleeve 221 or withdrawn from the magnetic rod sleeve 221.

[0062] In practical implementation, the turntable 1 can be a circular structure, and the column 24 can be set at the center of the turntable 1 to make full use of the space on the turntable 1 and reduce the space occupied by the module. Both the first drive unit and the second drive unit can be ball screw motors, which independently drive the first mounting bracket 22 and the second mounting bracket 21 to move along the axial direction of the turntable 1, that is, independently drive the first mounting bracket 22 and the second mounting bracket 21 to move up and down.

[0063] In specific implementation, a mounting base 23 is provided in the mounting through hole, and the turntable 1 can rotate relative to the mounting base 23. The column 24 and the liquid addition module 3 are located on the same side of the turntable 1 in the axial direction. One end of the column 24 is fixedly connected to the mounting base 23, so that the column 24 is installed in the center of the turntable 1.

[0064] Figures 11 and 12 show schematic diagrams of the liquid addition module of the protein purification apparatus provided in this application. As shown in Figures 11 and 12, the liquid addition module 3 may further include a liquid addition needle 31, which can be connected to a liquid addition pipeline to supply liquid to the liquid addition needle 31. The liquid addition needle 31 can move axially and radially along the turntable 1, and the liquid outlet is located on the liquid addition needle 31. In a specific implementation, the liquid addition module 3 may further include a third drive unit 32, a fourth drive unit 34, and a liquid addition needle holder 35. The third drive unit 32 is located on one side of the turntable 1 in the radial direction, and the third drive unit 32 may be a lead screw motor. The fourth drive unit 34 and the positioning seat 11 are located on the same side of the turntable 1 in the axial direction. The fourth drive unit 34 may also be a lead screw motor. The fourth drive unit 34 is connected to the third drive unit 32, and the third drive unit 32 can drive the fourth drive unit 34 to move axially along the turntable 1. Corresponding to the direction in the figure, that is, the third drive unit 32 can drive the fourth drive unit 34 to move up and down. The liquid injection needle holder 35 is connected to the fourth drive unit 34. The fourth drive unit 34 can drive the liquid injection needle holder 35 to move radially along the turntable 1. Corresponding to the direction in the figure, that is, the fourth drive unit 34 can drive the liquid injection needle holder 35 to move back and forth.

[0065] In actual setup, the dispensing needle 31 is mounted on the dispensing needle holder 35. Through the cooperation of the third drive unit 32 and the fourth drive unit 34, the dispensing needle 31 can move axially and radially along the turntable 1, that is, the dispensing needle 31 can move up and down and back and forth. In practical application, the dispensing needle 31 can be moved to the top of the container and aligned vertically with the container, and then moved downward into the container, thereby enabling the dispensing of liquid into the container.

[0066] In practice, the dispensing needle holder 35 is provided with multiple limiting holes, which are arranged in at least one row along the tangent of the turntable 1. One dispensing needle 31 is disposed in one limiting hole, and one dispensing needle 31 can add reagent to one container.

[0067] In a specific implementation, the liquid addition module 3 may further include a needle washing tank 33, which is mounted on the third drive unit 32. The needle washing tank 33 is located on the side of the liquid addition needle holder 35 facing the turntable 1, and has an opening on the side facing the liquid addition needle holder 35, through which the liquid addition needle 31 can extend into the needle washing tank 33. When it is necessary to change the type of reagent added to the container, the liquid addition needle 31 can first move above the needle washing tank 33, and then move downward into the needle washing tank 33. The cleaning solution and the new reagent pass through the liquid addition pipeline and the liquid addition pump in sequence and are discharged from the liquid addition needle 31, thereby rinsing the liquid addition needle 31 and achieving the purpose of cleaning the pipeline and washing the needle, which can avoid cross-contamination between different reagents. After the liquid addition needle 31 is rinsed, the new reagent is added to the container.

[0068] The following example illustrates the protein purification process using a turntable 1 comprising six positioning seats 11, forming six workstations (S, A, B, C, D, and E), with each workstation holding twelve containers (such as centrifuge tubes) and purifying magnetic beads of 1 mL or more. Workstation S can be selected as the sample position to hold samples, such as samples that have been incubated after adding magnetic beads. The other workstations can hold positioning racks 4 containing empty centrifuge tubes. The program can be set according to the expression level and volume of the samples to perform multiple operations, such as three washes and two elutions. Afterward, the purified protein solution is collected, and the magnetic beads are returned to the sample tubes.

[0069] (1) Place the twelve samples requiring purification with added magnetic beads into the positioning rack 4, and place the positioning rack 4 in position S. Place empty centrifuge tubes in the remaining positions. Install twelve new magnetic rod sleeves 221 on the first mounting rack 22, and install twelve magnetic rods 211 on the second mounting rack 21. Select the pre-set experimental program, click start, and the turntable 1 will rotate, moving position S below the mixing module 2. At this time, position A is below the liquid addition module 3.

[0070] (2) The magnetic rod 211 of the mixing module 2 moves down and extends into the magnetic rod sleeve 221. Then the magnetic rod 211 and the magnetic rod sleeve 221 move down together and extend into the centrifuge tube of the S station to adsorb the magnetic beads in the centrifuge tube.

[0071] (3) The liquid injection needle 31 of the liquid injection module 3 moves to the top of the centrifuge tube at station A, and the liquid injection needle 31 moves down and extends into the centrifuge tube at station A, and the first washing liquid is added according to the program settings.

[0072] (4) When the magnetic beads in the centrifuge tube at station S are adsorbed onto the magnetic rod sleeve 221, station A has completed automatic liquid addition, and the magnetic rod 211 and magnetic rod sleeve 221 move upward. Turntable 1 rotates clockwise one station (e.g., rotates 60°), at which point station A rotates to below the mixing module 2, and station B is below the liquid addition module 3, and the first washing liquid is automatically added to the centrifuge tube at station B.

[0073] (5) The magnetic rod 211 and the magnetic rod sleeve 221 move down and extend into the centrifuge tube at station A. The magnetic rod 211 moves up and exits the magnetic rod sleeve 221. The magnetic beads adsorbed on the magnetic rod sleeve 221 are released into the centrifuge tube at station A. The magnetic rod sleeve 221 moves up and down according to the amplitude and speed set in the program. After the magnetic beads are fully mixed with the reagents in the centrifuge tube at station A, the magnetic rod 211 moves down and extends into the magnetic rod sleeve 221 to adsorb the magnetic beads in the centrifuge tube at station A, thus completing the first cleaning.

[0074] (6) Turntable 1 continues to rotate clockwise by one station. At this time, station B is below the mixing module 2 and station C is below the liquid addition module 3. The liquid addition needle 31 moves to the needle washing tank 33 and is rinsed with the second washing liquid. Then the liquid addition needle 31 moves to the top of the centrifuge tube at station C and moves down into the centrifuge tube at station C. The second washing liquid is added according to the program settings.

[0075] (7) At this time, station B, which is located below the mixing module 2, repeats step (5) to complete the second washing of impurities.

[0076] (8) Turntable 1 continues to rotate clockwise one station. At this time, station C is below mixing module 2 and station D is below liquid addition module 3. Liquid addition needle 31 moves to needle washing tank 33 and uses cleaning solution and elution solution to rinse the pipeline and liquid addition needle 31. Then liquid addition needle 31 moves to the top of centrifuge tube at station D and moves down into centrifuge tube at station D. Elution solution is added according to the program settings.

[0077] (9) At this time, station C, which is located below the mixing module 2, repeats step (5) to complete the third washing of impurities.

[0078] (10) Turntable 1 continues to rotate clockwise by one station. At this time, station D is below mixing module 2 and station E is below liquid addition module 3. Liquid addition needle 31 moves down and extends into the centrifuge tube of station E, and eluent is added according to the program settings.

[0079] (11) At this time, station D, which is located below the mixing module 2, repeats step (5) to complete the first elution.

[0080] (12) Rotary disk 1 continues to rotate clockwise by one station. At this time, station E is below mixing module 2. Repeat step (5) to complete the second elution. At this time, the centrifuge tube at station E contains the final collection liquid.

[0081] (13) Turntable 1 continues to rotate clockwise one station, at which point station S is below mixing module 2. Magnetic rod 211 and magnetic rod sleeve 221 move down and extend into the centrifuge tube at station S. Then, magnetic rod 211 moves up, and magnetic rod sleeve 221 mixes at a set amplitude and speed, releasing the magnetic beads adsorbed on magnetic rod sleeve 221 into the centrifuge tube, and the process ends.

[0082] (14) Remove the collected liquid from the centrifuge tube at station E.

[0083] In the protein purification process described above, the washing buffer (first washing buffer and second washing buffer) can be PBS buffer or water, and the elution buffer can be an acidic solution (such as acetic acid).

[0084] The magnetic bead regeneration process will be explained below.

[0085] (1) Place the used magnetic beads in the centrifuge tube at station S, and place empty centrifuge tubes at the other stations. Select the regeneration process, and station S moves to below mixing module 2. Magnetic rod 211 and magnetic rod sleeve 221 move down and extend into the centrifuge tube at station S to adsorb the magnetic beads in the centrifuge tube. Magnetic rod 211 and magnetic rod sleeve 221 then move up. At this time, station A is below liquid addition module 3. Add magnetic bead regeneration buffer to the centrifuge tube at station A.

[0086] (2) Rotate turntable 1 clockwise by one station. At this time, station A is below mixing module 2. Magnetic rod 211 and magnetic rod sleeve 221 move down and are inserted into the centrifuge tube at station A. Magnetic rod 211 moves up, and the magnetic beads adsorbed on magnetic rod sleeve 221 are released into the centrifuge tube at station A. Magnetic rod sleeve 221 moves up and down slightly in the centrifuge tube. After the magnetic beads on magnetic rod sleeve 221 are fully mixed with the reagent in the centrifuge tube, magnetic rod 211 moves down and extends into magnetic rod sleeve 221 to adsorb the magnetic beads, thus completing one rinse. At this time, station B is below liquid addition module 3. Magnetic bead regeneration buffer is added to the centrifuge tube at station B.

[0087] (3) Turntable 1 continues to rotate clockwise one station. At this time, station B is below mixing module 2, and magnetic rod 211 and magnetic rod sleeve 221 move down to complete the second rinsing. Repeat the above actions to complete two more rinsing with washing buffer and one rinsing with storage buffer. Finally, the regenerated magnetic beads are stored in storage buffer.

[0088] In the magnetic bead regeneration process described above, the magnetic bead regeneration buffer can be an alkaline solution (such as NaOH aqueous solution), the washing buffer can be PBS buffer solution or water, and the storage buffer can be ethanol, etc.

[0089] As described above, the protein purification device provided in this application embodiment has multiple workstations, and the function of each workstation can be highly customized. The reagents added by the liquid addition module 3 are controlled according to a customized program, enabling operations such as incubation, mixing, liquid addition, washing, elution, regeneration, and magnetic bead recovery at each workstation. It achieves a high degree of automation with a small space footprint. It also supports customized single-sample parameters and can simultaneously process multiple (e.g., twelve) samples with different liquid addition parameters (twelve centrifuge tubes, each containing one sample). Furthermore, when the sample position rotates to the mixing module 2, the magnetic rod sleeve 221 descends and begins the mixing operation. At this time, the adjacent workstation is already automatically adding liquid under the liquid addition module 3, allowing for simultaneous mixing and automatic liquid addition, saving total protein purification time and improving purification efficiency.

[0090] Furthermore, by selecting magnetic rods 211 with different magnetic attraction forces, the purification needs of small (0.1-1 mL) and medium (1-8 mL) magnetic beads can be met, and the purification volume of magnetic beads covers a wide range (0.1-8 mL; the total volume of magnetic beads added to the sample).

[0091] Meanwhile, the sample incubation step before protein purification can be completed by the small up-and-down movement of the magnetic rod sleeve 221 of the mixing module 2, thereby achieving the binding of the magnetic beads with the target protein.

[0092] Furthermore, the processing solution generated at each step can be retained in the centrifuge tubes at each workstation, instead of being directly discharged into the waste liquid line. This facilitates later tracking to check for protein residue in the waste liquid, and the purification efficiency can be further improved by adjusting the volume of the added magnetic beads and the program settings. The liquid addition needle 31 is only used to add liquid to the centrifuge tubes and does not aspirate the processing solution from the centrifuge tubes back into the waste liquid line, thus avoiding contamination of the liquid addition needle 31 and preventing cross-contamination between samples from different centrifuge tubes.

[0093] The above are merely specific embodiments of this application, but the scope of protection of this application is not limited to the above embodiments. Obviously, those skilled in the art can make various modifications and variations to this application without departing from the scope and spirit of this application. If these modifications and variations fall within the scope of the claims of this application and their equivalents, then the intent of this application also includes these modifications and variations.

Claims

1. A protein purification apparatus, characterized in that, Includes a turntable, a liquid addition module, and a mixing module; The turntable has multiple positioning seats on one side in the axial direction. The multiple positioning seats are spaced apart along the circumference of the turntable. The positioning seats can rotate together with the turntable. The positioning seats are used to place at least one container. The liquid addition module and the positioning seat are located on the same side of the turntable in the axial direction; the liquid addition module includes at least one liquid outlet for adding reagents into the container; The mixing module and the liquid addition module are located on the same side of the turntable in the axial direction, and the mixing module and the liquid addition module are arranged at intervals along the circumference of the turntable; the mixing module includes a magnetic rod sleeve and a magnetic rod, and the magnetic rod is used to extend into the magnetic rod sleeve.

2. The protein purification apparatus according to claim 1, characterized in that, The plurality of positioning seats are arranged at equal intervals along the circumference of the turntable; Along the circumference of the turntable, the interval angle between the mixing module and the liquid addition module is the same as the interval angle between the adjacent positioning seats.

3. The protein purification apparatus according to claim 1 or 2, characterized in that, The mixing module further includes a first mounting bracket and a second mounting bracket, which are respectively capable of moving along the axial direction of the turntable. The projections of the first mounting bracket and the second mounting bracket at least partially overlap along the axial direction of the turntable. The magnetic rod sleeve is disposed on the side of the first mounting bracket away from the second mounting bracket, and the magnetic rod is disposed on the side of the second mounting bracket facing the first mounting bracket.

4. The protein purification apparatus according to any one of claims 1 to 3, characterized in that, The number of magnetic rod sleeves is multiple, and the multiple magnetic rod sleeves are arranged in at least one row along the tangent of the turntable, with each row of magnetic rod sleeves including at least one magnetic rod sleeve; The number of magnetic bars is multiple, and the multiple magnetic bars are arranged in at least one row along the tangent of the turntable, with each row of magnetic bars including at least one magnetic bar.

5. The protein purification apparatus according to claim 4, characterized in that, The number of magnetic rods is the same as the number of magnetic rod sleeves, and one magnetic rod is used to extend into one magnetic rod sleeve along the axial direction of the turntable.

6. The protein purification apparatus according to any one of claims 1 to 5, characterized in that, Along the axial direction of the turntable, the first end of the magnetic rod sleeve is provided with a insertion hole, the size of which is smaller than the size of the magnetic rod sleeve, and the projection of the magnetic rod is located within the insertion hole; the magnetic rod sleeve and the magnetic rod are respectively capable of moving along the axial direction of the turntable, the second end of the magnetic rod sleeve is used to extend into the container, and the magnetic rod is used to extend from the first end of the magnetic rod sleeve into the insertion hole to extend into the magnetic rod sleeve.

7. The protein purification apparatus according to claim 3, characterized in that, The turntable is provided with a mounting through hole, and the mounting through hole is coaxially arranged with the turntable. The mixing module also includes a column that extends along the axial direction of the turntable. The projection of the column on the axial direction of the turntable is located within the mounting through hole, and the turntable is rotatable relative to the column. The column is provided with a first driving part and a second driving part. The first mounting bracket is connected to the first driving part, and the first driving part is used to drive the first mounting bracket to move along the axial direction of the turntable. The second mounting bracket is connected to the second driving part, and the second driving part is used to drive the second mounting bracket to move along the axial direction of the turntable.

8. The protein purification apparatus according to claim 7, characterized in that, A mounting base is provided in the mounting through hole, and the turntable can rotate relative to the mounting base. The column and the liquid filling module are located on the same side of the turntable in the axial direction, and one end of the column is fixedly connected to the mounting base.

9. The protein purification apparatus according to any one of claims 1 to 8, characterized in that, The positioning seat is provided with a positioning frame, and the positioning frame is provided with at least one positioning hole. The axial direction of the positioning hole is parallel to the axial direction of the turntable, and the positioning hole is used to accommodate the container.

10. The protein purification apparatus according to any one of claims 1 to 9, characterized in that, The turntable is provided with multiple positioning protrusions, which form multiple positioning protrusion groups. The number of positioning protrusion groups is the same as the number of positioning seats. Each positioning protrusion group includes multiple positioning protrusions, and the multiple positioning protrusions in each positioning protrusion group are arranged around a positioning seat.

11. The protein purification apparatus according to any one of claims 1 to 10, characterized in that, The liquid filling module also includes a liquid filling needle, which is used to connect to the liquid filling pipeline. The liquid filling needle can move along the axial and radial directions of the turntable, and the liquid outlet is located on the liquid filling needle.

12. The protein purification apparatus according to claim 11, characterized in that, The liquid dispensing module further includes a third drive unit, a fourth drive unit, and a liquid dispensing needle holder, wherein the third drive unit is located on one side of the turntable in the radial direction; The fourth drive unit and the positioning seat are located on the same side of the turntable in the axial direction. The fourth drive unit is connected to the third drive unit, and the third drive unit can drive the fourth drive unit to move along the axial direction of the turntable. The liquid dispensing needle holder is connected to the fourth driving unit, which can drive the liquid dispensing needle holder to move radially along the turntable; The injection needle is mounted on the injection needle holder.

13. The protein purification apparatus according to claim 12, characterized in that, The liquid dispensing needle holder is provided with multiple limiting holes, and the multiple limiting holes are arranged in at least one row along the tangent of the turntable; One of the liquid injection needles is disposed within one of the limiting holes.

14. The protein purification apparatus according to claim 12 or 13, characterized in that, The liquid dispensing module also includes a needle washing tank, which is disposed on the third drive unit. The needle washing tank is located on the side of the liquid dispensing needle holder facing the turntable. The side of the needle washing tank facing the liquid dispensing needle holder has an opening, and the liquid dispensing needle is used to extend into the needle washing tank through the opening.

15. A method for purifying proteins using the protein purification apparatus according to any one of claims 1-14, characterized in that, Includes the following steps: 1) Place the positioning seat containing the container at one station of the turntable. The container contains a sample to be purified and the sample contains magnetic beads. The remaining stations of the turntable contain the positioning seat containing an empty container. 2) Control the magnetic rod sleeve of the mixing module to mix the sample in the container, and control the liquid addition module to add reagents to the empty container adjacent to the container in the rotation direction of the turntable; 3) Control the magnetic rod of the mixing module to magnetically attract the sample that has been mixed in the container; 4) Control the turntable to rotate one station, and control the magnetic rod of the mixing module to release the magnetically attracted sample into the container where the reagent has been added; 5) Repeat steps 2) to 4) until the entire setup process is complete.

16. The method as described in claim 15, characterized in that, The reagents added to the container by the liquid addition module are washing solution, elution solution, magnetic bead regeneration buffer, washing buffer or storage buffer.

17. The method as described in claim 16, characterized in that, Before replacing the reagent added to the container, the liquid dispensing module controls the liquid outlet to move to the needle washing tank, and rinses the liquid outlet with the replaced reagent, or simultaneously rinses the liquid dispensing pipeline connected to the liquid outlet.

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