A protein solution concentration device

Abstract

This invention discloses a protein solution concentration device, comprising: a concentration mechanism; a pressure mechanism; and fluid pipelines. The concentration mechanism includes: a first housing and a second housing, the first housing having an inlet for the protein solution to enter the concentration mechanism and a first outlet for the concentrated solution to exit the concentration mechanism; the second housing having a second outlet for waste liquid to exit the concentration mechanism; and an ultrafiltration membrane assembly disposed within the first and second housings. One side of the ultrafiltration membrane assembly has a first flow channel connecting the inlet to the outlet, and the other side of the ultrafiltration membrane assembly has a second flow channel connecting to the second outlet. With the above configuration, the concentration mechanism has a simple and practical structure, enabling rapid and efficient protein concentration while ensuring that the protein's activity and structure are not damaged. Simultaneously, the ultrafiltration membrane assembly of this invention exhibits low protein adhesion and high flux, avoiding waste due to protein adhesion during concentration.

Description

Technical Field

[0001] This invention relates to the field of protein purification in structural biology, and more particularly to a protein solution concentration apparatus. Background Technology

[0002] In structural biology protein experiments, protein expression and purification are relatively expensive. The initial volume of the protein solution to be concentrated is usually only a few milliliters to tens of milliliters, while the concentration factor is several to tens of times. Only the concentration of the target protein in the solution is concentrated. The concentration method needs to be fast and efficient to ensure that the activity and structure of the protein are not destroyed so that subsequent protein structure research can be carried out.

[0003] Ultrafiltration centrifugation is currently the most widely used protein concentration method in the field of structural biology protein purification. As a membrane separation technology, ultrafiltration is widely used for the concentration, desalting, and buffer exchange of biological samples. Its principle is that under the action of force, a solution passes through the pores of an ultrafiltration membrane. Large molecular weight particles are retained, while water, solvents, and low molecular weight solutes are allowed to permeate, thus achieving the purposes of concentration, desalting, and buffer exchange. The biggest advantages of this method are its simplicity, speed, and efficiency; it can simultaneously concentrate and purify molecules. Ultrafiltration centrifuge tubes are disposable ultrafiltration centrifugal filtration devices that can be used for protein sample concentration and protein buffer exchange. Compared with methods such as chromatography and dialysis, ultrafiltration is gentler on the molecules being processed, does not require organic extraction, does not cause protein denaturation, and is simple and efficient. However, conventional ultrafiltration centrifugation concentration leaves residual volume, leading to protein loss. Since the cost of obtaining protein samples is extremely high, protein loss due to residual volume is unacceptable. Meanwhile, centrifugation causes a gradient distribution of protein concentration within the ultrafiltration tube. Near the ultrafiltration membrane, there may be localized areas with excessively high protein concentrations, which can easily clog the ultrafiltration membrane. For some high-concentration, unstable proteins, this can lead to protein precipitation and denaturation, resulting in protein loss and concentration failure.

[0004] Chinese patent document CN212504858U discloses a protease extraction ultrafiltration device. It separates the protease from impurities by using a folded ultrafiltration belt within a square cylinder to filter out impurities in the solution. A secondary ultrafiltration operation is performed using a pressure tube and a second ultrafiltration component to further improve the extraction purity and efficiency of the protease. However, with prolonged use, impurities can adhere to the ultrafiltration belt, leading to a decrease in the extraction efficiency and purity of the protease.

[0005] Chinese patent document CN216303809U discloses a protease extraction ultrafiltration device, including an extraction chamber. A filter membrane is fixedly connected between the inner walls of the extraction chamber, dividing the interior of the extraction chamber into a stock solution chamber and an extract solution chamber. A turbulence-inducing mechanism is provided in the stock solution chamber, and an anti-clogging mechanism is provided in the extract solution chamber. A driving mechanism is located on one side of the extraction chamber. During filtration, the second shaft of the anti-clogging mechanism drives an eccentric wheel to rotate, which in turn drives a push plate and a sliding rod to slide. Multiple sliding rods work together to keep the filter membrane dynamic, preventing impurities from adhering to it. Furthermore, the turbulence-inducing mechanism allows the liquid and impurities on the surface of the filter membrane to flow slowly, preventing impurities from settling and accumulating on the membrane surface. However, this type of filter membrane is more susceptible to damage under internal tensile and compressive stress. Additionally, the molecular weight cutoff of the filter membrane is dynamically changing, resulting in a higher impurity content in the extract solution. Moreover, this structure is complex and occupies a large space. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention provides a protein solution concentration device with minimal protein loss.

[0007] A protein solution concentration apparatus, comprising:

[0008] Concentration mechanism, used to concentrate protein solutions;

[0009] Pressure mechanism, used to provide concentrated pressure for protein solutions;

[0010] Fluid piping is used to guide the protein solution into the protein solution concentration device and to connect the concentration mechanism and the pressure mechanism.

[0011] The concentration mechanism includes:

[0012] The first and second shells are covered. The first shell is provided with an inlet for the protein solution to enter the concentration mechanism and a first outlet for the concentrated liquid to flow out of the concentration mechanism. The second shell is provided with a second outlet for the waste liquid to flow out of the concentration mechanism.

[0013] Ultrafiltration membrane assembly disposed in the first housing and the second housing;

[0014] One side of the ultrafiltration membrane module is provided with a first flow channel that connects the inlet to the outlet, and the other side of the ultrafiltration membrane module is provided with a second flow channel that connects to the second outlet and whose flow path covers the flow path of the first flow channel.

[0015] Preferably, the concentration mechanism includes a stirring component, which is at least partially located in the first flow channel and used to stir the protein solution in the first flow channel, thereby promoting a more uniform concentration of the protein solution during the concentration process, and preventing problems such as protein adhesion, precipitation or blockage of the ultrafiltration membrane component caused by excessively high protein concentration near the ultrafiltration membrane component.

[0016] Furthermore, the concentration mechanism includes a substrate, an ultrafiltration membrane assembly, and a first flow plate located between the substrate and the ultrafiltration membrane assembly. The first flow plate has a first perforation extending through itself along its height direction. The substrate, the ultrafiltration membrane assembly, and the side of the first perforation constitute a first flow channel. The first flow channel is separately arranged to facilitate the arrangement of the stirring assembly in the first flow channel.

[0017] Furthermore, the stirring assembly includes a magnetic soft micropillar that can operate under changes in the magnetic field to achieve stirring. The magnetic soft micropillar has a simple structure and occupies little space, making it easy to place in the first flow channel to stir the protein solution.

[0018] Furthermore, the magnetic soft micropillars are cast onto a substrate after mixing magnetic materials with a flexible matrix. The magnetic materials include: iron(II,III) oxide, manganese-zinc ferrite, nickel-zinc ferrite, strontium ferrite, barium ferrite, etc.; the flexible matrix includes: polydimethylsiloxane, EVA resin, acrylic resin, polyacrylamide gel, sodium alginate gel, etc.; the mass ratio of flexible matrix to magnetic material is 4–19:1. This configuration results in a simple connection structure between the magnetic soft micropillars and the substrate, occupying minimal space, and does not negatively impact the flow and concentration of protein solutions or the ultrafiltration membrane assembly. In addition, the magnetic soft micropillars exhibit strong adhesion, reliably attaching to the substrate; simultaneously, they possess high flexibility, enabling efficient agitation of the protein solution under the action of a magnetic stirrer.

[0019] Furthermore, the first flow channel is a serpentine channel with a width of 0.01–1 mm and a height of 0.01–1 mm. The height of the magnetic soft microcolumn is 50%–80% of the height of the serpentine channel, and the diameter of the magnetic soft microcolumn is 10%–50% of the width of the serpentine channel. This configuration of the protein solution concentration device can be applied to the concentration of trace samples. The dimensions of the magnetic soft microcolumn and the serpentine channel are compatible. The magnetic soft microcolumn can efficiently agitate the protein solution while avoiding interference between the magnetic soft microcolumn and the serpentine channel during agitation.

[0020] Preferably, the protein solution concentration apparatus further includes: a first flow sensor for measuring the flow rate of the protein solution entering the concentration mechanism through the inlet; and / or a second flow sensor for measuring the flow rate of the concentrate flowing out of the concentration mechanism through the first outlet; and / or a third flow sensor for measuring the flow rate of the waste liquid flowing out of the concentration mechanism through the second outlet. The concentration factor of the protein solution is calculated based on the detection data from the first flow sensor, the second flow sensor, and the third flow sensor to improve the accuracy of the concentration factor calculation.

[0021] Preferably, the protein solution concentration apparatus further includes a collection mechanism for collecting the concentrate. The collection mechanism includes an ultraviolet detector for detecting the target protein, a controller, and a collector. The collector includes a first type of collection tube and a second type of collection tube. When the ultraviolet detector detects the presence of the target protein in the concentrate, the detector outputs a first signal, and the controller is in a first state, controlling the concentrate to flow into the first type of collection tube. When the ultraviolet detector detects the absence of the target protein in the concentrate, the detector outputs a second signal, and the controller is in a second state, controlling the concentrate to flow into the second type of collection tube. Through this configuration, the collection mechanism can distinguish whether the concentrate contains the target protein and can accurately collect concentrate containing the target protein, improving the reliability of the protein solution concentration apparatus.

[0022] Furthermore, the controller includes a support arm and an injection port. The support arm rotates and / or moves relative to the collector to switch between the first state and the second state. This configuration makes the controller simple and practical, and facilitates the controller to collect the concentrate into the first type of collection tube or the second type of collection tube depending on whether it has been concentrated to a preset concentration.

[0023] Furthermore, when the signal type output by the UV detector changes, there is a lag time in the state transition of the controller. The lag time is the ratio of the volume of the fluid pipeline from the UV detector to the injection port to the flow rate of the concentrate in the fluid pipeline, so that the concentrate in the fluid pipeline can flow completely into its corresponding collection tube before switching the collection tube, thereby avoiding waste of concentrate.

[0024] The beneficial effects of this invention are:

[0025] This invention provides a protein solution concentration apparatus; the concentrated protein solution has an accurate and controllable concentration; and the protein solution concentration apparatus provided by this invention is suitable for concentrating protein solutions with fixed volume and concentration, as well as for concentrating solutions with continuously injected, variable volume and protein concentration. The concentration mechanism of this invention has a simple and practical structure, enabling rapid and efficient protein concentration while ensuring that the protein's activity and structure are not damaged. Simultaneously, the ultrafiltration membrane component of this invention exhibits low protein adhesion and high flux, avoiding waste due to protein adhesion during concentration. Furthermore, using the protein solution concentration apparatus of this application can significantly improve the problems of ultrafiltration membrane clogging caused by excessively high local protein concentrations and the precipitation and denaturation of some proteins at high concentrations, reducing protein loss. Attached Figure Description

[0026] Figure 1 A three-dimensional structural diagram of the protein solution concentrate of the present invention.

[0027] Figure 2 An exploded view of the concentration mechanism of the present invention.

[0028] Figure 3 A three-dimensional structural diagram of the substrate and magnetic soft micropillars of the present invention.

[0029] Figure 4 A three-dimensional structural diagram of the collection mechanism of the present invention. Detailed Implementation

[0030] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] A protein solution concentration device 100 is disclosed. One end of the protein solution concentration device 100 can be connected to the sample outlet of a protein purification device, allowing the protein solution concentration device 100 to directly collect and concentrate the purified protein solution. This improves the convenience of protein purification and concentration, reduces errors from manual operation, and saves on manual operation costs. The other end of the protein solution concentration device 100 can also be connected to an injection device, enabling the protein solution concentration device 100 to be used to concentrate protein solutions of different types and applications, thus providing high flexibility and applicability.

[0032] like Figure 1As shown, the protein solution concentration device 100 includes a concentration mechanism 11, a pressure mechanism 12, a collection mechanism 13, and a fluid pipeline 14. The concentration mechanism 11, the pressure mechanism 12, and the collection mechanism 13 are connected through the fluid pipeline 14. After the protein solution enters the protein solution concentration device 100, it flows through the concentration mechanism 11 under the pressure of the pressure mechanism 12 and is concentrated by the concentration mechanism 11. Then it flows to the collection mechanism 13 to complete the collection.

[0033] like Figure 1 and Figure 2 As shown, the concentration mechanism 11 includes a first housing 111 and a second housing 112, which are mutually capped and sealed together. The first housing 111 is provided with an inlet 1111 and a first outlet 1112, and the second housing 112 is provided with a second outlet 1121. The protein solution enters the concentration mechanism 11 through the inlet 1111 and is concentrated to form a concentrated liquid and a waste liquid. The concentrated liquid flows out of the concentration mechanism 11 through the first outlet 1112, and the waste liquid flows out of the concentration mechanism 11 through the second outlet 1121.

[0034] Specifically, an ultrafiltration membrane assembly 113 is disposed in the first housing 111 and the second housing 112. The ultrafiltration membrane assembly 113 is capable of retaining molecules smaller than a preset molecular weight. As an alternative implementation, the ultrafiltration membrane assembly 113 can be made of polyethersulfone membrane to reduce the low protein adhesion of the ultrafiltration membrane assembly 113 while giving the ultrafiltration membrane assembly 113 a high flux.

[0035] A first flow channel 101 is provided on one side of the ultrafiltration membrane assembly 113, connecting the inlet 1111 to the first outlet 1112; a second flow channel 102 is provided on the other side of the ultrafiltration membrane assembly 113, at least partially connecting to the second outlet 1121, and the flow path of the second flow channel 102 can cover the flow path of the first flow channel 101. When the protein solution enters the concentration mechanism 11 through the inlet 1111 and flows in the first flow channel 101, under the pressure of the pressure mechanism 12, impurities and some liquid in the protein solution pass through the ultrafiltration membrane assembly 113 and enter the second flow channel 102 to form waste liquid. The waste liquid flows in the second flow channel 102 and flows out of the concentration mechanism 11 through the second outlet 1121; the remaining protein solution in the first flow channel 101 forms a concentrated solution and flows out of the concentration mechanism 11 through the first outlet 1112.

[0036] More specifically, the concentration mechanism 11 includes a substrate 114 and a first flow plate 115. The substrate 114 is located between the first flow plate 115 and the first housing 111. The substrate 114 and the first housing 111 are sealed together or integrally formed. The substrate 114 and the first flow plate 115 are sealed together. The ultrafiltration membrane assembly 113 is located on the side of the first flow plate 115 away from the substrate 114 and is at least partially sealed together with the first flow plate 115. Along the height direction of the first flow plate 115, the first flow plate 115 is provided with a first perforation 1151 that penetrates itself. The substrate 114, the ultrafiltration membrane assembly 113, and the side of the first perforation 1151 surround each other to form a first flow channel 101. A first through hole 1141 is provided on the substrate 114 at the positions corresponding to the liquid inlet 1111 and the first liquid outlet 1112. The protein solution enters the first flow channel 101 through the liquid inlet 1111 and the first through hole 1141, and the concentrate flows out of the first flow channel 101 through the first through hole 1141 and the first liquid outlet 1112.

[0037] The concentration mechanism 11 includes a cover plate 116 and a second flow plate 117. The cover plate 116 is located between the second flow plate 117 and the second housing 112. The cover plate 116 and the second housing 112 are sealed together or integrally formed. The cover plate 116 and the second flow plate 117 are also sealed together. The ultrafiltration membrane assembly 113 is located on the side of the second flow plate 117 away from the cover plate 116 and is at least partially sealed together with the second flow plate 117. Along the height direction of the second flow plate 117, a second perforated portion 1171 is provided on the second flow plate 117. The sides of the cover plate 116, the ultrafiltration membrane assembly 113, and the second perforated portion 1171 surround each other to form a second flow channel 102. A second through hole 1161 is provided on the cover plate 116 at the position corresponding to the second liquid outlet 1121. Waste liquid flows out of the second flow channel 102 through the second through hole 1161 and the second liquid outlet 1121.

[0038] like Figure 1 and Figure 3 As shown, the concentration mechanism 11 also includes a stirring component 118, which is at least partially located in the first flow channel 101. The stirring component 118 agitates the protein solution in the first flow channel 101, making the concentration of the protein solution more uniform during the concentration process. At the same time, it can also prevent problems such as protein adhesion, precipitation or blockage of the ultrafiltration membrane component 113 caused by excessively high protein concentration near the ultrafiltration membrane component 113.

[0039] Specifically, the stirring assembly 118 includes magnetic soft micropillars 1181, which are located on the end face of the substrate 114 away from the first housing 111. The magnetic soft micropillars 1181 are uniformly distributed in the first flow channel 101 and can move under changing magnetic fields to achieve stirring. As an alternative implementation, a magnetic stirrer 200 is provided on one side of the concentration mechanism 11. The magnetic stirrer 200 generates a changing magnetic field, and the magnetic soft micropillars 1181 stir the protein solution under the action of the magnetic stirrer 200. As an alternative implementation, the substrate 114 can be made of polymers such as polydimethylsiloxane, polyethylene, or polypropylene, or it can be made of glass, quartz, or silicon wafers, to facilitate microelectromechanical processing or injection molding of the substrate 114; the magnetic soft micropillars 1181 are formed by mixing magnetic materials and flexible matrix and then casting them onto the substrate 114 through a mold. Preferably, the magnetic material is iron(III) oxide nanomaterial, the flexible matrix is ​​polydimethylsiloxane, and the mass ratio of the flexible matrix to the magnetic material is 4 to 19:1. Because the contact area between the magnetic soft micropillar 1181 and the substrate 114 is small and they are far from the magnetic stirrer 200, the connection structure between the magnetic soft micropillar 1181 and the substrate 114 in this configuration is simple and occupies little space. It will not have a negative impact on the flow and concentration of the protein solution or on the ultrafiltration membrane assembly 113. In addition, the magnetic soft micropillar 1181 has strong adhesion and can be firmly attached to the substrate 114. At the same time, the magnetic soft micropillar 1181 has strong flexibility and can efficiently stir the protein solution under the action of the magnetic stirrer 200.

[0040] As an alternative implementation, the flow path of the first flow channel 101 and the flow path of the second flow channel 102 are basically the same, that is, the structures of the first flow plate 115 and the second flow plate 117 are basically the same, so as to facilitate the manufacturing of the first flow plate 115 and the second flow plate 117 and save design costs.

[0041] As an alternative implementation, both the first flow channel 101 and the second flow channel 102 can be serpentine channels to increase the contact area between the protein solution and the ultrafiltration membrane assembly 113, thereby improving the stirring effect of the magnetic soft microcolumn 1181 on the protein solution. Simultaneously, it facilitates support for the ultrafiltration membrane assembly 113, thus improving the concentration effect of the ultrafiltration membrane assembly 113. The width of the serpentine channel is 0.01–1 mm, and the height is 0.01–1 mm. The height of the magnetic soft microcolumn 1181 is 50%–80% of the height of the serpentine channel, and the diameter of the magnetic soft microcolumn 1181 is 10%–50% of the width of the serpentine channel. This configuration of the protein solution concentration device 100 can be applied to the concentration of trace samples. The size matching between the magnetic soft microcolumn 1181 and the serpentine channel allows the magnetic soft microcolumn 1181 to efficiently stir the protein solution while avoiding interference between the magnetic soft microcolumn 1181 and the serpentine channel during stirring. The magnetic soft micropillar 1181 can be cylindrical or prismatic, etc. It is understood that the second flow channel 102 does not necessarily have to have the same shape as the first flow channel 101. Any technical solution where the flow path of the second flow channel 102 covers the flow path of the first flow channel 101 and the ultrafiltration membrane assembly 113 is supported is within the scope of protection of this invention.

[0042] like Figure 1 As shown, the protein solution concentration device 100 also includes a first flow sensor 15 and a second flow sensor 16. The first flow sensor 15 is connected to the inlet 1111 via a fluid pipeline 14 and detects the flow rate of the protein solution in real time. The second flow sensor 16 is connected to the first outlet 1112 via a fluid pipeline 14 and detects the flow rate of the concentrate in real time. Based on the detection data from the first flow sensor 15 and the second flow sensor 16, the concentration factor of the protein solution can be calculated. The pressure value generated by the pressure mechanism 12 is adjusted according to the difference between the calculated concentration factor and the desired concentration factor, thereby controlling the flow rate of the concentrate and the flow rate of the waste liquid. This realizes the control function of the concentration factor by the protein solution concentration device 100, and makes the control of the concentration factor by the protein solution concentration device 100 highly accurate and reliable. As an alternative implementation, the protein solution concentration apparatus 100 further includes a third flow sensor 17. The third flow sensor 17 is connected to the second outlet 1121 via a fluid conduit 14 and monitors the flow rate of the waste liquid in real time. The concentration factor of the protein solution is calculated jointly based on the detection data from the first flow sensor 15, the second flow sensor 16, and the third flow sensor 17, thereby improving the accuracy of the concentration factor calculation. Optionally, the protein solution concentration apparatus 100 may use at least two of the first flow sensor 15, the second flow sensor 16, and the third flow sensor 17 to calculate the concentration factor of the protein solution.

[0043] like Figure 1and Figure 4 As shown, the collection mechanism 13 includes an ultraviolet detector 131, a controller 132, and a collector 133. The concentration of the target protein in the fraction output from the protein purification device is not fixed. Before the target protein peak, the fraction is a buffer solution without the target protein; during the target protein peak, the fraction contains a protein solution; and after the target protein peak, the fraction contains a buffer solution without the target protein. The ultraviolet detector 131 can determine the concentration of the target protein in the concentrate based on the ultraviolet absorption intensity of the concentrate. Based on the target protein concentration detected by the ultraviolet detector 131, the pressure value generated by the pressure mechanism 12 is calculated and dynamically adjusted to ensure that the concentration of the target protein in the concentrate meets the preset requirements. The controller 132 includes a first state and a second state, and the collector 133 includes a first type of collection tube 1331 and a second type of collection tube 1332. When the UV detector 131 detects the presence of the target protein in the concentrate, it outputs a first signal. At this time, the controller 132 is in a first state, controlling the concentrate to flow into the first type of collection tube 1331. When the UV detector 131 detects the absence of the target protein in the concentrate, it outputs a second signal. At this time, the controller 132 is in a second state, controlling the concentrate to flow into the second type of collection tube 1332. Through this configuration, the collection mechanism 13 can distinguish whether the concentrate contains the target protein and can accurately collect concentrate that meets the preset requirements, improving the reliability of the protein solution concentration device 100.

[0044] Specifically, the collector 133 also includes a base 1333, which is a cylindrical structure that can rotate around its own axis and whose end face is close to the collector 133. The end face of the base 1333 close to the collector 133 is provided with a first type of receiving hole 13331 and a second type of receiving hole 13332. The first type of receiving hole 13331 is evenly distributed on a circle with a first radius around the axis of the base 1333, and the first type of collecting tube 1331 is located in the first type of receiving hole 13331. The second type of receiving hole 13332 is evenly distributed on a circle with a second radius around the axis of the base 1333, and the second type of collecting tube 1332 is located in the second type of receiving hole 13332. The controller 132 also includes a support arm 1321 and an injection port 1322 located on the support arm 1321. The support arm 1321 is rotatable and / or movable relative to the base 1333, such that when the controller 132 is in a first state, the axis of the injection port 1322 is substantially coincident with the axis of the first type of receiving orifice 13331; when the controller 132 is in a second state, the axis of the injection port 1322 is substantially coincident with the axis of the second type of receiving orifice 13332. The base 1333 rotates around its own axis, facilitating the collection of the concentrate by the collector 133. The controller 132's structure is simple and practical, allowing it to easily collect the concentrate into the first type of collection tube 1331 or the second type of collection tube 1332 depending on whether it contains the target protein.

[0045] It should be noted that the initial concentration of the target protein in the protein solution output from the injection device is fixed. The protein solution concentration device 100 concentrates the protein solution and collects it in the first collection tube 1331. After injection, protein residue will remain in the protein solution concentration device 100. Therefore, after injection, a certain volume of buffer solution needs to be injected into the protein solution concentration device 100 to further push and concentrate the remaining protein, and collect it in the first collection tube 1331. This ensures that there is no protein residue in the protein solution concentration device 100 and no protein loss during the concentration process.

[0046] As an alternative implementation, when the signal type output by the UV detector 131 changes, the state transition of the controller 132 has a lag time to allow the concentrate in the fluid line 14 to completely flow into its corresponding collection tube before switching the collection tube, thereby avoiding waste of the concentrate. The lag time is calculated as follows: Lag time = Volume of the fluid line 14 from the UV detector 131 to the injection port 1322 / Flow rate of the concentrate in the fluid line 14.

[0047] This invention provides a protein solution concentration device 100 with almost no protein damage. The concentrated protein solution has an accurate and controllable concentration. The protein solution concentration device 100 provided by this invention is suitable for concentrating protein solutions with fixed volume and concentration, as well as for concentrating continuously injected protein solutions with variable volume and concentration. Furthermore, by incorporating a magnetic stirrer 200 and magnetic soft micropillars 1181 in the concentration mechanism 11, and by molding the magnetic soft micropillars 1181 from a mixture of magnetic material and flexible matrix onto a substrate 114 using a mold, the connection structure between the magnetic soft micropillars 1181 and the substrate 114 is simple and occupies little space. This arrangement does not negatively impact the flow and concentration of the protein solution or the ultrafiltration membrane assembly 113. It also ensures that the magnetic soft micropillars 1181 have strong adhesion, allowing them to reliably adhere to the substrate 114, and possess strong flexibility, enabling efficient agitation of the protein solution under the action of the magnetic stirrer 200. The protein solution concentration device 100, through the first flow sensor 15, the second flow sensor 16, and the third flow sensor 17, can control the concentration factor, ensuring high accuracy and reliability in this control. The collection mechanism 13, using the ultraviolet detector 131, can distinguish whether the concentrate contains the target protein and accurately collect concentrate containing the target protein, further improving the reliability of the protein solution concentration device 100.

[0048] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A protein solution concentration apparatus, comprising: Concentration mechanism, used to concentrate protein solutions; Pressure mechanism, used to provide concentrated pressure for protein solutions; A fluid conduit is provided for guiding the protein solution into the protein solution concentration device and for connecting the concentration mechanism and the pressure mechanism. Its features are, The concentration mechanism includes: The first and second housings are covered. The first housing is provided with an inlet for the protein solution to enter the concentration mechanism and a first outlet for the concentrated liquid to flow out of the concentration mechanism. The second housing is provided with a second outlet for the waste liquid to flow out of the concentration mechanism. An ultrafiltration membrane assembly disposed in the first housing and the second housing; One side of the ultrafiltration membrane assembly is provided with a first flow channel that connects the inlet to the outlet, and the other side of the ultrafiltration membrane assembly is provided with a second flow channel that connects to the second outlet and whose flow path covers the flow path of the first flow channel. The concentration mechanism includes a stirring assembly, which is at least partially located in the first flow channel and is used to stir the protein solution in the first flow channel. The stirring assembly includes magnetic soft micropillars that can operate under changes in the magnetic field to achieve stirring; The first flow channel is a serpentine channel with a width of 0.01–1 mm and a height of 0.01–1 mm. The height of the magnetic soft micropillar is 50% to 80% of the height of the serpentine channel, and the diameter of the magnetic soft micropillar is 10% to 50% of the width of the serpentine channel.

2. The protein solution concentration apparatus according to claim 1, characterized in that, The concentration mechanism includes a substrate, an ultrafiltration membrane assembly, and a first flow plate located between the substrate and the ultrafiltration membrane assembly. The first flow plate has a first perforation extending through itself along its height direction. The substrate, the ultrafiltration membrane assembly, and the side of the first perforation constitute the first flow channel.

3. The protein solution concentration apparatus according to claim 2, characterized in that, The magnetic soft micropillars are formed by casting a mixture of magnetic material and flexible matrix onto a substrate. The magnetic material is at least one of the following: iron(II,III) oxide, manganese-zinc ferrite, nickel-zinc ferrite, strontium ferrite, and barium ferrite. The flexible matrix is ​​at least one of the following: polydimethylsiloxane, EVA resin, acrylic resin, polyacrylamide gel, and sodium alginate gel; The mass ratio of the flexible matrix to the magnetic material is 4 to 19:

1.

4. The protein solution concentration apparatus according to claim 1, characterized in that, Also includes: A first flow sensor that measures the flow rate of the protein solution entering the concentration mechanism through the inlet; And / or a second flow sensor that measures the flow rate of the concentrate flowing out of the concentration mechanism through the first outlet; And / or a third flow sensor that measures the flow rate of waste liquid exiting the concentration mechanism through the second outlet.

5. The protein solution concentration apparatus according to claim 1, characterized in that, The protein solution concentration device further includes a collection mechanism for collecting the concentrate. The collection mechanism includes an ultraviolet detector for detecting the target protein, a controller, and a collector. The collector includes a first type of collection tube and a second type of collection tube. When the ultraviolet detector detects that the concentrate contains the target protein, the ultraviolet detector outputs a first signal. At this time, the controller is in a first state, and the controller controls the concentrate to flow into the first type of collection tube. When the ultraviolet detector detects that the concentrate does not contain the target protein, the ultraviolet detector outputs a second signal. At this time, the controller is in a second state, and the controller controls the concentrate to flow into the second type of collection tube.

6. The protein solution concentration apparatus according to claim 5, characterized in that, The controller includes a support arm and an injection port, the support arm rotating and / or moving relative to the collector to achieve the transition between the first state and the second state.

7. The protein solution concentration apparatus according to claim 5, characterized in that, When the signal type output by the ultraviolet detector changes, there is a lag time in the state transition of the controller. The lag time is the ratio of the volume of the fluid pipeline from the ultraviolet detector to the injection port to the flow rate of the concentrate in the fluid pipeline.

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