Purification apparatus for preparing nucleic acid drugs

Abstract

The application relates to the technical field of pharmaceutical equipment, and provides a purification equipment for preparing nucleic acid drugs, which comprises a first purification process module, the first purification process module comprises a first separation and purification equipment, a first mixed container, a first displacement liquid input end and a pump assembly, the first separation and purification equipment has a first inlet end, a first outlet end and a first waste liquid outlet end, the first separation and purification equipment is arranged in a first circulation loop, the first circulation loop is provided with a first purification input end, the first mixed container is arranged in the first circulation loop, and the inlet of the first mixed container is connected to the first outlet end and the outlet is connected to the first inlet end, the first displacement liquid input end is connected to the inlet of the first mixed container, and the pump assembly is used for driving solution flow. Through the arrangement of the first purification process module, effective purification of the solution can be realized. After the solution is introduced into the first circulation loop, the solution can flow repeatedly in the first circulation loop alone and / or together with other auxiliary liquids, so that purification is realized.

Description

Technical Field

[0001] This application relates to the field of pharmaceutical equipment technology, specifically to a purification device for preparing nucleic acid drugs. Background Technology

[0002] mRNA (messenger RNA) is a single-stranded ribonucleic acid that carries genetic information (DNA transcription products). Its main function is to accurately transcribe the genetic information from DNA and translate it into proteins on ribosomes. mRNA stock solutions are pharmaceutical preparations made with mRNA as the core component. They regulate protein expression by transmitting genetic information and are currently mainly used in vaccine development, gene therapy, and protein replacement therapy.

[0003] Currently, the production process of mRNA stock solutions requires a lot of manual intervention, which not only results in low production efficiency but also makes the solution susceptible to contamination during the transfer process. Therefore, a purification device for preparing nucleic acid drugs is proposed to address these issues. Utility Model Content

[0004] This application provides a purification device for preparing nucleic acid drugs, thereby improving the preparation effect of nucleic acid drugs.

[0005] This application provides an apparatus for preparing nucleic acid drugs, including a first purification process module, the first purification process module comprising, The first separation and purification device has a first inlet end, a first outlet end and a first waste liquid outlet end. The first separation and purification device is configured in a first circulation loop, and the first circulation loop has a first purification input end. A first mixing container is configured in the first circulation loop, with its inlet connected to the first outlet and its outlet connected to the first inlet. The first displacement fluid inlet is connected to the inlet of the first mixing container; and Pump assembly used to drive the flow of solution.

[0006] In some embodiments, the first purification process module further includes, A first purification and collection end is connected to the first mixing container; and A first filtration component is disposed between the first mixing container and the first purification collection end.

[0007] In some embodiments, the first purification process module further includes, The first gas inlet is connected to the first filter assembly; and The first detection sensor is used to detect pressure changes in the pipeline where the first filter assembly is located when gas is input into the first filter assembly at the first gas inlet end.

[0008] In some embodiments, the first purification process module further includes, At least one first pre-prepared inlet; and a second waste liquid collection end; The first separation and purification device is configured via a pipeline between the first pre-preparation inlet and the second waste liquid collection end.

[0009] In some embodiments, the first mixing container includes... Container body; The inlet pipe extends into the container body and then extends toward the side wall of the container body; An outlet pipe extends to the bottom of the container body; and The stirring component includes a stirring drive mechanism and a stirring part that are connected in a mating manner, with the stirring part located within the container body.

[0010] In some embodiments, the first mixing container is disposed on a mixing mechanism, the mixing mechanism comprising: Mixing drive component; An eccentric connector is eccentrically connected to the mixing drive. The support member is rotatably connected to the eccentric connector; When the mixing drive unit drives the eccentric connector to move, it can cause the support and the first mixing container to rotate and shake, so as to mix the medicine liquid in the first mixing container.

[0011] In some embodiments, the second purification process module includes, The second separation and purification device has a second inlet end, a second outlet end and a second waste liquid outlet end. The second separation and purification device is configured in a second circulation loop. The second circulation loop has a second purification input end. The second mixing container is configured in the second circulation loop, and its inlet is connected to the second outlet end and its outlet is connected to the second inlet end; At least two auxiliary liquid inlets are connected to the inlet of the first mixing container; and Pump assembly used to drive the flow of solution.

[0012] In some embodiments, the second purification process module further includes, The second purification and collection end is connected to the second mixing container; and The second filtration device is disposed between the second mixing container and the second purification collection end.

[0013] In some embodiments, at least one set of pre-purification process modules is provided upstream of the first purification process module.

[0014] In some embodiments, the pre-purification process module includes a chromatography process module, the chromatography process module including, The chromatography main pipeline includes a first port and a second port for fluid connection, as well as a third port and a fourth port for fluid connection; The chromatography inlet pipe is connected to the first port; A chromatography column, the inlet of which is connected to the second port and the outlet of which is connected to the third port; The chromatography outlet pipe is connected to the fourth port.

[0015] In some embodiments, the first port may be selectively connected to the third port, and the second port may be selectively connected to the fourth port.

[0016] In some embodiments, the first port may be selectively connected to the fourth port.

[0017] In some embodiments, a bubble sensor is provided in the chromatography inlet line; the apparatus for preparing nucleic acid drugs further includes a bubble removal line, which is connected to the chromatography inlet line via a bubble trap.

[0018] In some embodiments, when two or more chromatography inlet lines are provided, the two or more chromatography inlet lines are connected together to a static mixer and then to the chromatography main line, the static mixer having a bent flow channel.

[0019] In some embodiments, the pre-purification process module includes a magnetic bead purification process module, which includes, At least one magnetic bead is used for purification at the inlet end; A magnetic bead purification container, with its inlet and outlet connected by pipes to form a magnetic bead purification circulation loop; and A magnetic source, located at the magnetic bead purification container, is used to selectively adsorb magnetic beads.

[0020] In some embodiments, an oscillation mechanism is further included, connected to the magnetic bead purification container, and drives the solution and magnetic beads within the magnetic bead purification container to oscillate.

[0021] In some embodiments, the magnetic source includes a magnetic body and a magnetic source driving mechanism, the magnetic source driving mechanism being connected to the magnetic body to drive the magnetic body to move between positions of adsorption and desorption.

[0022] In some embodiments, the purification device for preparing nucleic acid drugs further includes at least one mounting bracket, the mounting bracket including a frame, and a plurality of first connectors and a plurality of second connectors disposed on the frame, wherein the first connectors extend along a first direction, the second connectors extend along a second direction, and the first and second directions are alternately arranged.

[0023] In some embodiments, the first direction is a vertical direction, the second direction is a horizontal direction, and the bottom end of the first connector is also provided with a hook structure.

[0024] This application provides the following beneficial effects: It offers a purification device for preparing nucleic acid drugs, which, through the setup of a first purification process module, enables effective purification of the solution. After the solution is introduced into the first circulation loop, it can flow repeatedly in the first circulation loop, either alone or together with other auxiliary solutions, thereby achieving purification.

[0025] In addition, the first purification process module can be combined and worked together with the second purification process module, the pre-purification process module, etc., to achieve better purification results.

[0026] It can also work in conjunction with other components to enable the preparation of nucleic acid drugs. Attached Figure Description

[0027] Figure 1 An exemplary schematic diagram of an in vitro transcription process module is shown.

[0028] Figure 2 An exemplary top view of an in vitro transcription process module is shown.

[0029] Figure 3 An exemplary schematic diagram of a transfer unit is shown.

[0030] Figure 4 Example shown Figure 2 A magnified view of part A in the diagram.

[0031] Figure 5 An exemplary schematic diagram of a hybrid unit is shown.

[0032] Figure 6 An exemplary schematic diagram of a hybrid drive mechanism is shown.

[0033] Figure 7 An exemplary schematic diagram of an insulated container is shown.

[0034] Figure 8 An example is shown. Figure 7 Top half-section view.

[0035] Figure 9Example shown Figure 8 A schematic diagram of the PP cross-section.

[0036] Figure 10 An exemplary schematic diagram of a buffer container and a base in conjunction is shown.

[0037] Figure 11 An exemplary schematic diagram of a buffer container is shown.

[0038] Figure 12 An exemplary schematic diagram of a base for adapting a buffer container is shown.

[0039] Figure 13 An exemplary schematic diagram of the steps of an in vitro transcription method is shown.

[0040] Figure 14 An exemplary diagram of the piping for the first purification process module is shown.

[0041] Figure 15 An exemplary piping diagram of the second purification process module is shown.

[0042] Figure 16 An exemplary schematic diagram of the mixing bottle is shown.

[0043] Figure 17 An exemplary piping diagram of a chromatography process module is shown.

[0044] Figure 18 An exemplary schematic diagram of a magnetic bead purification process module is shown.

[0045] Figure 19 An exemplary piping diagram of a magnetic bead purification process module is shown.

[0046] Figure 20 An exemplary schematic diagram of a mixing mechanism is shown.

[0047] Figure 21 An exemplary top view of a mixing mechanism is shown.

[0048] Figure 22 Example shown Figure 21 A sectional view of section AA.

[0049] Figure 23 An exemplary structural schematic diagram of an eccentric connector is shown.

[0050] Figure 24 An exemplary schematic diagram of a static mixer is shown.

[0051] Figure 25 An exemplary top half-sectional view of a static mixer is shown.

[0052] Figure 26 An exemplary schematic diagram of an installation frame is shown.

[0053] Figure 27 An exemplary flowchart of a method for preparing nucleic acid drugs is shown.

[0054] Explanation of reference numerals in the figure: 11-Working platform, 12-Reaction dish, 100-Transfer unit, 110-First transfer drive mechanism, 111-First lateral movement mechanism, 112-First rotation mechanism, 113-First vertical movement mechanism, 120-First transfer component, 121-Pipette tip, 122-Pressure source, 123-Tip mounting part, 130-Second transfer drive mechanism, 131-Second lateral movement mechanism, 132-Second rotation mechanism, 133 - Second vertical moving mechanism, 140-Second transfer component, 150-Clamping mechanism, 151-Clamping component, 160-Refrigeration chamber, 170-Waste liquid chamber, 180-Buffer container, 1801-Magnetic rotor, 1802-Second positioning structure, 181-Base, 1811-Magnetic drive component, 1812-First positioning structure, 1813-Weighing assembly, 191-Refrigerated preparation storage area, 192-Ambient temperature preparation storage area, 193-Pipette tip storage area, 20 0-Mixing unit, 210-Bearing component, 211-Insulated base, 212-Insulated container, 213-Insulated cover, 214-Elastic component, 220-Mixing drive mechanism, 221-Power source, 222-Drive eccentric wheel, 223-First eccentric transmission component, 224-Passive eccentric wheel, 225-Second eccentric transmission component, 226-Base, 240-Tilting assembly, 241-Tilting motor, 242-Reducer, 243-Connecting frame, 244-Positioning assembly. 300 - Chromatography process module; 310 - Chromatography inlet pipeline; 320 - Chromatography main pipeline; 321 - First port; 322 - Second port; 323 - Third port; 324 - Fourth port; 330 - Chromatography outlet pipeline; 340 - Waste liquid collection container; 350 - Static mixer; 351 - Flow channel; 352 - Body; 353 - Reagent inlet; 354 ​​- Reagent outlet; 355 - Sealing cap; 356 - Sealing ring; 360 - Bubble removal pipeline.361 - Bubble Trap; 400 - Magnetic Bead Purification Process Module; 410 - Magnetic Bead Purification Inlet; 420 - Magnetic Bead Purification Container; 430 - Exhaust End; 440 - Waste Liquid Outlet; 450 - Magnetic Bead Purification Outlet; 460 - Magnetic Source; 500 - First Purification Process Module; 510 - First Purification Input; 520 - First Separation and Purification Device; 521 - First Inlet; 522 - First Outlet; 523 - First Waste Liquid Outlet; 524 - First Waste... 525 - First mixing container; 526 - First replacement fluid input; 530 - First purification collection end; 531 - First filter assembly; 532 - First bubble sensor; 540 - First pre-preparation inlet; 541 - Second waste liquid collection end; 542 - First sensing assembly; 550 - First gas inlet; 551 - First detection sensor; 700 - Second purification process module; 710 - Second purification input; 720 - Second separation and purification device 721-Second Inlet, 722-Second Outlet, 723-Second Waste Liquid Outlet, 724-Third Waste Liquid Collection, 725-Second Mixing Container, 726-Auxiliary Liquid Inlet, 730-Second Purification Collection, 731-Second Filter Component, 732-Second Bubble Sensor, 740-Second Preparatory Inlet, 741-Fourth Waste Liquid Collection, 742-Second Sensing Component, 750-Second Gas Inlet, 751-Second Detection Sensor 760-Metering auxiliary component, 810-Container body, 820-Inlet pipe, 830-Outlet pipe, 840-Agitator, 910-Mixing drive component, 920-Eccentric connector, 921-Shaft, 922-Base, 930-Bearing, 940-Support component, 950-Flexible wall, 960-Position sensor, 970-Weight sensor, 1001-Frame, 1002-First connector, 1003-Second connector, 1004-Hook structure. Detailed Implementation

[0055] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application. In addition, it should be understood that the specific embodiments described herein are only for illustration and explanation of this application and are not intended to limit this application. In this application, unless otherwise stated, directional terms such as "up," "down," "left," and "right" generally refer to up, down, left, and right in the actual use or working state of the device, specifically the drawing directions in the accompanying drawings.

[0056] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined with "first" or "second" may explicitly or implicitly include at least one of that feature.

[0057] This application provides an apparatus and a method for preparing nucleic acid drugs, which are described in detail below. It should be noted that the order of description of the following embodiments is not intended to limit the preferred order of the embodiments of this application. Furthermore, the descriptions of each embodiment have their own emphasis; parts not described in detail in a certain embodiment can be referred to in the relevant descriptions of other embodiments.

[0058] This application provides an apparatus for preparing nucleic acid drugs, including a first purification process module 500 (see [link]). Figure 14 ), lipid nanoparticle preparation process module and second purification process module 700 (see also...) Figure 15 ).

[0059] The first purification process module 500 is used to purify the incoming drug solution; A lipid nanoparticle preparation process module is located downstream of the first purification process module 500 for preparing lipid nanoparticles; and The second purification process module 700 is located downstream of the lipid nanoparticle preparation process module to purify the solution containing lipid nanoparticles.

[0060] Here, by way of example, each process module is equipped with a corresponding disposable consumable component, and the upstream disposable consumable component is connected to the downstream disposable consumable component via a sterile connection method. For example, the disposable consumable component may include pipes and / or pipe connection structures, etc.

[0061] An automatic control system can also be set up here. The automatic control system is connected to the first purification process module 500, the lipid nanoparticle preparation process module and the second purification process module 700 by signal connection. In this way, the automatic control system can realize the linkage control of the first purification process module 500, the lipid nanoparticle preparation process module and the second purification process module 700.

[0062] The automatic control system can be a central processing unit (CPU), microcontroller (MCU), digital signal processor (DSP), field-programmable gate array (FPGA), application-specific integrated circuit (ASIC), or other devices with data processing capabilities.

[0063] For example, the automatic control system can control each process module to carry out the corresponding process flow according to the needs of different processes and formulas, and transport the raw liquid obtained by the upstream process module to the downstream process module, and make each process module process the raw liquid in sequence to obtain the desired product.

[0064] The lipid nanoparticle preparation process module uses existing equipment. For example, CN120242853A discloses a nanomedicine preparation system and process, which can be used as a lipid nanoparticle preparation process module. Of course, the lipid nanoparticle preparation process module used in this application is not necessarily the same as that one. This is only an example of the structure of that disclosure text. There are various lipid nanoparticle preparation process modules in the prior art.

[0065] In some embodiments, please refer to Figure 14 The first purification process module 500 includes a hollow fiber column for performing purification operations. It is understood that a hollow fiber column has been used as an example here; however, in other embodiments, the hollow fiber column may be replaced with other first separation and purification devices 520, such as membrane packs, and the examples in this application do not constitute an undue limitation.

[0066] Exemplarily, the first inlet end 521 and the first outlet end 522 of the hollow fiber column are interconnected by a pipeline to form a first circulation loop, and the first circulation loop has a first purification input end 510 for inputting a solution from the upstream component. For example, the first inlet end 521 and the first outlet end 522 may be located at the bottom and top of the hollow fiber column, respectively, and a first waste liquid outlet end 523 is also provided on one side of the hollow fiber column, which may be connected to a first waste liquid collection end 524. Of course, this example does not constitute an undue limitation on this application, and in other embodiments, the positions of the first inlet end 521 and the first outlet end 522 may also be different.

[0067] In use, the solution from the upstream component, such as the solution from the in vitro transcription device, is input into the first circulation loop and then into the hollow fiber column for concentration. The waste liquid is discharged through the first waste liquid outlet 523 to the first waste liquid collection end 524, which may be equipped with a waste liquid collection container, etc.

[0068] For example, a displacement fluid is required during the purification operation, so a first displacement fluid input terminal 526 is provided, and a first mixing container 525 is provided in the aforementioned first circulation loop. For example, the inlet of the first mixing container 525 is connected to the first displacement fluid input terminal 526 and the first outlet terminal 522 of the aforementioned hollow fiber column, and the outlet of the first mixing container 525 is connected to the first inlet terminal 521 of the aforementioned hollow fiber column.

[0069] Therefore, the solution from the upstream equipment, or the solution and displacement fluid after the aforementioned concentration operation, can be input into the first mixing container 525. Furthermore, after the displacement fluid is added, the solution and displacement fluid are fed together into the hollow fiber column through the first circulation loop for cyclic concentration, and the waste liquid is discharged through the first waste liquid outlet 523 to the first waste liquid collection container 340.

[0070] Therefore, in this embodiment, the solution from the upstream is first concentrated by circulating it through a hollow fiber column, and then the concentrated solution and the replacement solution are concentrated together by circulating them, thereby achieving a better purification effect.

[0071] Of course, in some embodiments, the solution from upstream and the replacement fluid can be directly circulated and concentrated together, but in this case, some impurities may not be eliminated.

[0072] In some embodiments, a sensor may be installed in the pipeline between the first waste liquid outlet end 523 and the first waste liquid collection end 524 of the hollow fiber column. For example, a flow sensor may be installed. When the reading of the flow sensor reaches a preset value, it indicates that the discharged waste liquid has reached a preset amount, that is, the preset purification requirement has been met, and the circulation concentration stops. Alternatively, a UV sensor may be installed. When the reading of the UV sensor is within a preset range, it indicates that the concentration of the discharged waste liquid meets the requirements, that is, the preset purification requirement has been met, and the circulation concentration stops. Of course, in some examples, multiple sensors may be installed simultaneously. When the readings of multiple sensors all meet the preset requirements, it is determined that the preset purification requirement has been met, and the circulation concentration stops.

[0073] In some embodiments, the first displacement fluid inlet 526 is also connected to the first outlet 522 of the hollow fiber column, so as to subsequently push out the drug solution in the hollow fiber column and / or nearby pipelines through the displacement fluid.

[0074] In some embodiments, the first purification process module 500 further includes a first purification collection end 530, which is connected to the aforementioned first mixing container 525 to facilitate the collection of the purified solution to the first purification collection end 530. Exemplarily, various first filtration components 531, such as a butterfly filter or a capsule filter, can be provided between the first purification collection end 530 and the first mixing container 525 to achieve sterile filtration.

[0075] For example, a first bubble sensor 532 may also be provided upstream of the first filter assembly 531, which stops the collection of solution when bubbles are detected.

[0076] In addition, a purification and collection container may be provided at the first purification and collection end 530, or it may be directly connected to other downstream components, etc. The examples in this application embodiment do not impose undue limitations on it.

[0077] In some embodiments, at least one first pre-preparation inlet 540 and a second waste liquid collection end 541 may be provided. The hollow fiber column is disposed between the first pre-preparation inlet 540 and the second waste liquid collection end 541. The first pre-preparation inlet 540 is used to input the solution required for the pre-preparation stage into the hollow fiber column and other components to perform the pre-preparation steps. For example, the pre-preparation steps may include pure water cleaning, equilibrium liquid rinsing, alkaline solution rinsing, etc.

[0078] For example, three first pre-preparation inlet ends 540 are provided here. In use, the three first pre-preparation inlet ends 540 can be used to input different solutions to achieve the pre-preparation step. The waste liquid formed after the different solutions pass through the first inlet end 521 and the first outlet end 522 of the hollow fiber column in sequence flows to the second waste liquid collection end 541 and is discharged.

[0079] For example, the first pre-preparation inlet 540 inputs pure water, which flows through the pipeline and components within it to achieve pure water cleaning; the second pre-preparation inlet 540 inputs balancing fluid, which flows through the pipeline and components within it to achieve balancing fluid rinsing, and the balancing fluid can mainly be used to calibrate the sensors in the pipeline; the third pre-preparation inlet 540 inputs alkaline solution, which flows through the pipeline and components within it to achieve alkaline solution rinsing.

[0080] For example, a first sensing component 542 may be provided between the first pre-preparation inlet 540 and the second waste liquid collection end 541, and / or a first sensing component 542 may also be provided between the first pre-preparation inlet 540 and the first waste liquid collection end 524. The first sensing component 542 is used to detect various parameter values ​​of the solution. For example, the first sensing component 542 may include any of the following: a flow sensor, a mass sensor, a pH sensor, a UV value sensor, etc. Furthermore, the first sensing component 542 may be located downstream of the hollow fiber column. Also, the aforementioned equilibration liquid may be used to calibrate the first sensing component 542; that is, the state of the first sensing component 542 is determined by the reading of the first sensing component 542 as the equilibration liquid flows through it.

[0081] In some embodiments, at least one first pre-preparation inlet 540 is connected to the aforementioned first filter assembly 531 for rinsing the first filter assembly 531.

[0082] For example, a first pre-preparation inlet 540, a first filter assembly 531, and a second waste liquid collection end 541 are connected in sequence via pipelines.

[0083] In some embodiments, a first gas inlet 550 is also provided, which is connected to the aforementioned first filter assembly 531. Thus, by inputting gas, a first detection sensor 551, such as a pressure sensor, can detect whether the first filter assembly 531 is damaged. This is because when the membrane or other structure of the first filter assembly 531 is damaged, the reading of the first detection sensor 551 when gas passes through will differ from the reading under normal conditions.

[0084] For example, the first gas inlet 550, the first detection sensor 551, the first filter assembly, and the second waste liquid collection end 541 are connected in sequence through pipelines.

[0085] It is understood that, in addition, several pump assemblies configured in various pipelines can be provided in the first purification process module 500 to drive the solution and allow it to flow in each pipeline. The pump assemblies can be peristaltic pumps, etc. For example, the pump assemblies provided in this embodiment may include P1, P2, and P3. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the pump assemblies may also employ other configuration methods, locations, and quantities.

[0086] It is understood that, in addition, several valve assemblies configured in various pipelines can be provided in the first purification process module 500 to control the opening and closing of the pipelines. Exemplarily, the valve assemblies provided in the embodiments of this application may include V1 to V16. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the valve assemblies may also adopt other configuration methods, locations, and quantities, for example, being configured in various components that provide and collect solutions.

[0087] Therefore, based on the first purification process module 500 provided in the embodiments of this application, a pipeline setup and corresponding structure for purifying solutions are provided. On this basis, those skilled in the art can easily add corresponding pump components and / or valve components according to actual needs, thereby achieving the required control functions.

[0088] In some embodiments, please refer to Figure 15 The second purification process module 700 includes a hollow fiber column for performing purification operations.

[0089] Here, a hollow fiber column is used as an example for illustration. However, it is understood that in other embodiments, the hollow fiber column may be replaced with other second separation and purification devices 720, such as membrane packs. The examples in this application do not constitute an undue limitation on them.

[0090] Exemplarily, the second inlet end 721 and the second outlet end 722 of the hollow fiber column are interconnected by a pipeline to form a second circulation loop, and the second circulation loop has a second purification input end 710 for inputting a solution from the upstream component. For example, the second inlet end 721 and the second outlet end 722 can be located at the bottom and top ends of the hollow fiber column, respectively, and a second waste liquid outlet end 723 is also provided on one side of the hollow fiber column, which can be connected to a third waste liquid collection end 724. Of course, this example does not constitute an undue limitation of this application, and in other embodiments, the positions of the second inlet end 721 and the second outlet end 722, etc., may also be different.

[0091] In use, the solution from the upstream component, such as the solution from the lipid nanoparticle preparation process module, is input into the second circulation loop and then into the hollow fiber column for concentration. The waste liquid is discharged through the second waste liquid outlet 723 to the third waste liquid collection end 724, which may be equipped with a waste liquid collection container, etc.

[0092] For example, some auxiliary liquids are required during purification operations, such as displacement fluid, mother liquor, and auxiliary material mother liquor. The amount of auxiliary liquid required may vary depending on the actual process requirements.

[0093] Therefore, at least two auxiliary liquid input terminals 726 can also be provided here for inputting auxiliary liquids such as replacement fluid, mother liquor, and excipient mother liquor. In the figure, an embodiment with three auxiliary liquid input terminals 726 is shown as an example.

[0094] Furthermore, a second mixing container 725 is provided in the aforementioned second circulation loop. For example, the inlet of the second mixing container 725 is connected to the auxiliary liquid input end 726 and the second outlet end 722 of the aforementioned hollow fiber column, and the outlet of the second mixing container 725 is connected to the second inlet end 721 of the aforementioned hollow fiber column.

[0095] Therefore, the solution from the upstream equipment, or the solution and auxiliary liquid after the aforementioned concentration operation, can be fed into the second mixing container 725. Furthermore, after adding at least one auxiliary liquid, the solution and auxiliary liquid are fed together into the hollow fiber column through the second circulation loop for cyclic concentration, and the waste liquid is discharged through the second waste liquid outlet 723 to the second waste liquid collection container 340.

[0096] Therefore, in this embodiment, the solution from the upstream is first concentrated by circulating it through a hollow fiber column, and then the concentrated solution and the auxiliary liquid are concentrated together by circulating it, thereby achieving a better purification effect.

[0097] Of course, in some embodiments, the solution and auxiliary liquid from the upstream can be directly circulated and concentrated together, but in this case, some impurities may not be eliminated.

[0098] In some embodiments, in order to calculate the amount of auxiliary liquid more accurately, a metering component may be provided at the auxiliary liquid input end 726. For example, the metering component may be a weighing component, which can weigh the weight change of the auxiliary liquid container containing the auxiliary liquid in real time, thereby determining the amount of auxiliary liquid added.

[0099] Because of the presence of pipelines, although metering components such as weighing devices can determine the amount of auxiliary liquid added, some of the auxiliary liquid actually remains in the pipeline and does not completely enter the second mixing container 725. Therefore, in order to more accurately measure the amount of auxiliary liquid added, a metering auxiliary component 760 can also be provided. The metering auxiliary component 760 is disposed in the pipeline to measure the flow rate of the auxiliary liquid flowing through the pipeline.

[0100] For example, a weighing component can be installed at the auxiliary liquid inlet 726 as a metering component; a bubble sensor or flow sensor can be installed in the pipeline between the auxiliary liquid inlet 726 and the second mixing container 725 as a metering auxiliary component 760. During initial input, before the auxiliary liquid reaches the metering auxiliary component 760, only the existing air in the pipeline flows through it, and the metering auxiliary component 760 does not generate a reading until the auxiliary liquid reaches it. Therefore, only when both the metering component and the metering auxiliary component 760 reach their preset reading ranges is it confirmed that sufficient auxiliary liquid has been added.

[0101] Furthermore, in another scenario, such as when one auxiliary solution is added first and then another, the later-added auxiliary solution needs to push the previously added auxiliary solution forward before it can enter the second mixing container 725. In this case, the metering auxiliary component 760 is continuously generating readings, but the actually added auxiliary solution has not yet flowed into the second mixing container 725.

[0102] In this case, a more accurate addition amount can be determined by combining the liquid capacity in the calculation pipeline, the readings of the metering component, and the metering auxiliary component 760. For example, it would be more accurate to use the subsequent reading as the amount of auxiliary liquid added by subtracting the pre-calculated liquid capacity in the pipeline from the reading of the metering auxiliary component 760, based on the pre-calculated liquid capacity in the pipeline. Furthermore, the actual addition amount can also be determined by subtracting the pre-calculated capacity of the pipeline from the auxiliary liquid inlet 726 to the metering auxiliary component 760 or the second mixing container 725 from the reading of the metering component.

[0103] As another example, the metering auxiliary component 760 can be positioned near the second mixing container 725. For instance, multiple auxiliary liquid inlets 726 are interconnected, connected to the metering auxiliary component 760, and then connected to the second mixing container 725.

[0104] In some embodiments, a sensor may also be installed in the pipeline between the second waste liquid outlet end 723 and the second waste liquid collection end 724 of the hollow fiber column. For example, a flow sensor may be installed. When the reading of the flow sensor reaches a preset value, it indicates that the discharged waste liquid has reached a preset amount, that is, the preset purification requirement has been met, and the cycle concentration stops. Alternatively, a UV sensor may be installed. When the reading of the UV sensor is within a preset range, it indicates that the concentration of the discharged waste liquid meets the requirements, that is, the preset purification requirement has been met, and the cycle concentration stops. Of course, in some examples, multiple sensors may be installed simultaneously. When the readings of multiple sensors all meet the preset requirements, it is determined that the preset purification requirement has been met, and the cycle concentration stops.

[0105] In some embodiments, the auxiliary liquid inlet 726 is also connected to the second outlet 722 of the hollow fiber column, so as to subsequently push out the drug solution in the hollow fiber column and / or nearby pipelines by the replacement liquid.

[0106] In some embodiments, the second purification process module 700 further includes a second purification collection end 730, which is connected to the aforementioned second mixing container 725 to collect the purified solution to the second purification collection end 730. Exemplarily, various second filtration components 731, such as a butterfly filter or a capsule filter, can be provided between the second purification collection end 730 and the second mixing container 725 to achieve sterile filtration.

[0107] For example, a second bubble sensor 732 may also be provided upstream of the second filter assembly 731, which stops the collection of solution when bubbles are detected.

[0108] In addition, a purification and collection container may be provided at the second purification and collection end 730, or it may be directly connected to other downstream components, etc. The examples in this application embodiment do not impose undue limitations on it.

[0109] In some embodiments, at least one second pre-preparation inlet 740 and a fourth waste liquid collection end 741 may be provided. The hollow fiber column is disposed between the second pre-preparation inlet 740 and the fourth waste liquid collection end 741. The second pre-preparation inlet 740 is used to input the solution required for the pre-preparation stage into the hollow fiber column and other components to perform the pre-preparation steps. For example, the pre-preparation steps may include pure water cleaning, equilibrium liquid rinsing, alkaline solution rinsing, etc.

[0110] For example, three second pre-preparation inlet ends 740 are provided here. In use, the three second pre-preparation inlet ends 740 can be used to input different solutions to achieve the pre-preparation step. The waste liquid formed after the different solutions pass through the second inlet end 721 and the second outlet end 722 of the hollow fiber column flows to the fourth waste liquid collection end 741 and is discharged.

[0111] For example, the second pre-preparation inlet 740 inputs pure water, which flows through the pipeline and components within it to achieve pure water cleaning; the second pre-preparation inlet 740 inputs balancing fluid, which flows through the pipeline and components within it to achieve balancing fluid rinsing, and the balancing fluid can mainly be used to calibrate the sensors in the pipeline; the third pre-preparation inlet 740 inputs alkaline solution, which flows through the pipeline and components within it to achieve alkaline solution rinsing.

[0112] As another example, a second sensing component 742 may be provided between the second pre-preparation inlet 740 and the fourth waste liquid collection end 741, and / or a second sensing component 742 may also be provided between the second pre-preparation inlet 740 and the fourth waste liquid collection end 741. The second sensing component 742 is used to detect various parameter values ​​of the solution. For example, the second sensing component 742 may include any of the following: a flow sensor, a mass sensor, a pH sensor, a UV value sensor, etc. Furthermore, the second sensing component 742 may be located downstream of the hollow fiber column. Also, the aforementioned equilibration liquid may be used to calibrate the second sensing component 742; that is, the state of the second sensing component 742 is determined by the reading of the second sensing component 742 as the equilibration liquid flows through it.

[0113] In some embodiments, at least one second pre-preparation inlet 740 is connected to the aforementioned second filter assembly 731 for rinsing the second filter assembly 731.

[0114] For example, a second pre-prepared inlet 740, a second filter assembly 731, and a second sensing assembly 742 are connected in sequence via pipelines.

[0115] In some embodiments, a first gas inlet 550 is also provided, which is connected to the aforementioned second filter assembly 731. Thus, by inputting gas, a second detection sensor 751, such as a pressure sensor, can detect whether the second filter is damaged. This is because when the membrane or other structure of the second filter is damaged, the reading of the second detection sensor 751 when gas passes through will differ from the reading under normal conditions.

[0116] For example, the second gas inlet 750, the second detection sensor 751, the second filter assembly 731 and the fourth waste liquid collection end 741 are connected in sequence through pipelines.

[0117] It is understood that, in addition, several pump assemblies configured in various pipelines can be provided in the second purification process module 700 to drive the solution and allow it to flow in each pipeline. The pump assemblies can be peristaltic pumps, etc. For example, the pump assemblies provided in this embodiment may include P4, P5, and P6. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the pump assemblies may also employ other configuration methods, locations, and quantities.

[0118] It is understood that, in addition, several valve assemblies configured in various pipelines can be provided in the second purification process module 700 to control the on / off state of the pipelines. Exemplarily, the valve assemblies provided in the embodiments of this application may include V17 to V33. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the valve assemblies may also adopt other configuration methods, locations, and quantities, for example, being configured in various components that provide and collect solutions.

[0119] Therefore, based on the second purification process module 700 provided in the embodiments of this application, a pipeline setup and corresponding structure for purifying solutions are provided. Furthermore, those skilled in the art can easily add corresponding pump components and / or valve components according to actual needs, thereby achieving the required control functions.

[0120] In some embodiments, please refer to Figure 16 Either the first mixing container 525 or the second mixing container 725 may include a container body, an inlet pipe, an outlet pipe, and a stirring element. The inlet pipe extends into the container body and then extends towards the side wall of the container body, while the outlet pipe extends to the bottom of the container body. The stirring element includes a cooperating stirring drive mechanism and a stirring section, which is located within the container body for stirring and mixing the solution.

[0121] In other embodiments, either the first mixing container 525 or the second mixing container 725 described above can be mounted on the mixing mechanism. Driven by the mixing mechanism, the first mixing container 525 or the second mixing container 725 can be moved to achieve mixing of the medicinal liquid inside the first mixing container 525 or the second mixing container 725.

[0122] Please see Figure 20 The mixing mechanism may include a mixing drive 910 and an eccentric connector 920. The eccentric connector 920 is eccentrically connected to the mixing drive 910 and can move eccentrically under the drive of the mixing drive 910. The support member 940 is connected to the eccentric connector 920 and can also be driven by the eccentric connector 920 to move eccentrically. See here. Figure 21 and Figure 22The support member 940 is used to accommodate the aforementioned first mixing container 525 or second mixing container 725. The first mixing container 525 and / or the second mixing container 725 are mixing bags. The support member 940 is provided with an installation groove, which can accommodate and fix the first mixing container 525 or the second mixing container 725. In addition, the connection between the two can be reinforced with straps or the like. Of course, the example in this embodiment does not constitute an improper limitation.

[0123] Furthermore, in some embodiments, the support member 940 may be rotatably connected to the eccentric connector 920, for example, the eccentric connector 920 having a shaft portion 921, please refer to... Figure 23 A shaft portion 921 is mounted on a base 922, and a support member 940 is rotatably connected to the shaft portion 921 via a bearing 930. Alternatively, the shaft portion 921 may be inclined to a horizontal plane. Thus, when the mixing drive member 910 drives the eccentric connector 920, the support member 940 rotatably connected to the shaft portion 921 rotates and wobbles, thereby mixing the first mixing container 525 or the second mixing container 725 mounted thereon.

[0124] In further embodiments, please refer to... Figure 22 Furthermore, a flexible wall 950 can be provided circumferentially on the underside of the support member 940. The flexible wall 950 can be made of materials such as rubber. As the support member 940 moves, the flexible wall 950 can deform accordingly. The space formed by the flexible wall 950 is not easily invaded by external dust and components.

[0125] In a further embodiment, a position sensor 960 may be provided on one side of the eccentric connector 920. The position sensor 960 can be used to identify the position of the support member 940. For example, it can be used to identify the number of rotations of the support member 940.

[0126] In a further embodiment, a weight sensor 970 may be provided on the lower side of the support 940. The weight sensor 970 is used for weighing, so that the mass of the liquid medicine in the first mixing container 525 or the second mixing container 725 can be measured.

[0127] In some embodiments, at least one set of pre-purification process modules may be provided upstream of the first purification process module 500.

[0128] In some examples, please refer to Figure 17The pre-purification process module includes a chromatography process module 300, which includes a chromatography pipeline, a chromatography column (not shown) disposed within the chromatography pipeline, and a pump assembly for driving the movement of the solution within the chromatography pipeline. The chromatography pipeline includes a chromatography inlet pipeline 310, a chromatography outlet pipeline 330, and a chromatography main pipeline 320. The chromatography main pipeline 320 includes a first port 321 and a second port 322 for fluid connection, and a third port 323 and a fourth port 324 for fluid connection.

[0129] The first port 321 is connected to the chromatography inlet pipe 310, the second port 322 and the third port 323 are connected to the inlet and outlet of the chromatography column, respectively, and the fourth port 324 is connected to the chromatography outlet pipe 330. Thus, after the solution enters the chromatography line through the chromatography inlet pipe 310, it can flow to the chromatography column through the first port 321 and the second port 322, and flow to the chromatography outlet pipe 330 through the third port 323 and the fourth port 324.

[0130] For example, the first port 321 may be selectively connected to the third port 323, and the second port 322 may be selectively connected to the fourth port 324. Thus, the solution may enter the chromatography line through the chromatography inlet line 310, flow to the chromatography column through the first port 321 and the third port 323, and flow to the chromatography outlet line 330 through the second port 322 and the fourth port 324.

[0131] As another example, the first port 321 may be selectively connected to the fourth port 324, thereby allowing the solution to flow directly from the chromatography inlet line 310 through the first port 321 and the fourth port 324 into the chromatography outlet line 330 in some cases. For example, during calibration, cleaning, or other operations of the chromatography process module 300, the liquid may flow directly through this passage.

[0132] In some embodiments, a sensing component is configured in the chromatography pipeline. To achieve different functions, the sensing component may include any of the following: a bubble sensor, a pressure sensor, a flow sensor, a mass sensor, a pH sensor, and a UV sensor. For example, a bubble sensor, a pressure sensor, and a flow sensor may be configured in the chromatography inlet pipeline 310. As another example, a pH sensor and a UV sensor may be configured in the chromatography outlet pipeline 330.

[0133] In some embodiments, based on the example in which a bubble sensor is provided in the chromatography pipeline, the chromatography pipeline further includes a bubble removal pipeline 360, which is connected to the chromatography inlet pipeline 310 via a bubble trap 361.

[0134] It is understandable that valves can be installed between the various ports, thereby enabling selective connection or disconnection between the ports.

[0135] The chromatography inlet pipe 310 is configured as two pipes, with inlet ends A1 and A2 in one pipe and inlet ends B1 and B2 in the other. However, in other embodiments, the chromatography inlet pipe 310 may have more or fewer inlet ends, and / or, in other embodiments, the chromatography inlet pipe 310 may have one or other numbers of pipes. The examples in the embodiments of this application do not constitute an undue limitation.

[0136] In embodiments with two or more chromatography inlet lines 310, these inlet lines 310 are connected to a static mixer 350. The static mixer 350 has at least one bend in the flow channel 351, whereby when various different pharmaceutical solutions flow into the static mixer 350, the solutions collide with the wall of the bend in the flow channel 351, thereby promoting mixing. Exemplarily, to achieve better mixing, the flow channel 351 may have at least two bends.

[0137] This static mixer 350 can achieve mixing effects without power.

[0138] For example, please refer to Figure 24 and Figure 25 The static mixer 350 also includes a body 352, which has a liquid inlet 353 and a liquid outlet 354. A flow channel 351 is disposed on the body 352, and the flow channel 351 fluidly connects the liquid inlet 353 and the liquid outlet 354. Furthermore, a sealing cap 355 is provided on the body 352 to cooperate with the body 352 and close the flow channel 351. As another example, a sealing ring 356 may also be provided between the flow channel 351 and the sealing cap 355.

[0139] The chromatography outlet pipe 330 is configured as a single line with outlet ends C1 and C2. Outlet end C1 is connected to the waste liquid collection container 340, and outlet end C2 can be used to output the chromatographically purified solution, which can be connected to other downstream pipes or components. However, in other embodiments, the chromatography inlet pipe 310 may have more outlet ends, and / or, in other embodiments, the chromatography outlet pipe 330 may have multiple lines. The examples in the embodiments of this application do not constitute an undue limitation.

[0140] It is understood that, in addition, the chromatography process module 300 may also include several pump assemblies configured in various pipelines to drive the solution and allow the solution to flow in the various pipelines. The pump assemblies may be peristaltic pumps, etc. For example, the pump assemblies provided in this embodiment may include P9 and P10. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the pump assemblies may also employ other configurations, locations, and quantities.

[0141] It is understood that, in addition, the chromatography process module 300 may also include several valve assemblies configured in various pipelines to control the on / off state of the pipelines. Exemplarily, the valve assemblies provided in the embodiments of this application may include V41 to V57. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the valve assemblies may also employ other configuration methods, locations, and numbers, for example, being configured in various components that provide and collect solutions.

[0142] In other examples, please refer to Figure 18 and Figure 19 The pre-purification process module includes a magnetic bead purification process module 400. The magnetic bead purification process module 400 can utilize specific ligands modified on the surface of magnetic beads to bind with target molecules, and use a magnetic field to separate the magnetic beads from impurities, ultimately releasing a high-purity sample.

[0143] Here, the magnetic bead purification process module 400 includes at least one magnetic bead purification inlet 410 and a magnetic bead purification container 420. The magnetic bead purification inlet 410 is fluidly connected to the magnetic bead purification container 420, so that the solution can be input into the magnetic bead purification container 420 through the magnetic bead purification inlet 410.

[0144] The magnetic bead purification inlet 410 is used to input solutions from upstream and various other required solutions, such as solutions from the in vitro transcription device, as well as elution buffers and washing buffers. Multiple magnetic bead purification inlets 410 can be provided, or multiple solutions can be shared; this application does not impose undue limitations on this. For example, three inlets can be provided to respectively input the solution used for in vitro transcription, the elution buffer, and the washing buffer. Furthermore, in some examples, a storage container for storing solutions can be provided at the magnetic bead purification inlet 410.

[0145] For example, please refer to Figure 19 The inlet and outlet of the magnetic bead purification container 420 can be connected by pipelines to form a magnetic bead purification circulation loop, thereby allowing the solution to be circulated multiple times to achieve better magnetic bead purification effect.

[0146] As another example, in addition to the inlet and outlet for supplying solution, the magnetic bead purification container 420 may also be provided with an exhaust port, which is connected to an exhaust end 430. Furthermore, a gas collection container or the like may be provided at the exhaust end 430.

[0147] During use, the gas in the pipeline and various components can be discharged through the exhaust port.

[0148] In some examples, a liquid sensor may also be installed on the pipeline between the vent and the vent end 430.

[0149] When the magnetic bead purification process module 400 starts working, driven by the pump assembly and other components, the solution begins to enter the pipeline and move towards the magnetic bead purification container 420. Gas in the pipeline and the magnetic bead purification container 420 is expelled and exits through the exhaust port. Once the magnetic bead purification container 420 is nearly full, the solution begins to flow out from the exhaust port. When the liquid sensor detects the solution, it indicates that venting is complete, and the valve assembly can close the pipeline between the exhaust port and the exhaust end 430.

[0150] In addition, a magnetic source 460 is provided at the magnetic bead purification container 420. The magnetic source 460 can be a magnet or the like, and is used to selectively adsorb magnetic beads. When the magnetic beads in the solution pass through the magnetic bead purification container 420, the magnetic source 460 can apply a magnetic force to adsorb the magnetic beads and retain them in the magnetic bead purification container 420.

[0151] Here, the outlet of the magnetic bead purification container 420 is also connected to a waste liquid outlet 440, which may be equipped with a collection container. After the desired components are combined, for example, after the solution passes through the magnetic bead purification container 420 or through the magnetic bead purification loop multiple times, the solution is discharged to the waste liquid outlet 440.

[0152] The magnetic beads are then washed by introducing washing solution into the magnetic bead purification container 420. Here, the magnetic source 460 selectively adsorbs and desorbs the magnetic beads, thereby enabling the magnetic beads to achieve a better washing effect in the washing solution.

[0153] For example, the magnetic source 460 is movably positioned, thereby enabling selective adsorption and desorption by moving the position of the magnetic source 460. For example, the magnetic source 460 is directly or indirectly connected to a magnetic source 460 driving mechanism, such as a motor, which drives the magnetic source 460 to move, thereby moving it between the adsorption and desorption positions.

[0154] As another example, the magnetic bead purification container 420 is also provided with an oscillation mechanism, such as an oscillation motor, so that when the adsorption is desorbed, an oscillation action is applied to the magnetic bead purification container 420, so that the magnetic beads can be better washed in the washing liquid.

[0155] After washing is completed, the washing liquid is discharged to the waste liquid outlet at 440.

[0156] A magnetic bead purification outlet 450 is also provided here, which is connected to the outlet of the magnetic bead purification container 420. For example, the active ingredients bound on the magnetic beads can be eluted into the solution by introducing eluent through the magnetic bead purification inlet 410, and then the solution is transported to other downstream components through the magnetic bead purification outlet 450, such as the first purification process module 500 downstream.

[0157] Furthermore, the magnetic beads in the aforementioned solution can be automatically added to the solution by means of a robotic arm or by setting up a magnetic bead supply pipeline connected to the magnetic bead purification process module 400, or they can be added manually. The examples in this application embodiment do not impose undue limitations on this.

[0158] It is understood that, in addition, the magnetic bead purification process module 400 may also include several pump assemblies configured in various pipelines to drive the solution and allow the solution to flow in each pipeline. The pump assemblies may be peristaltic pumps, etc. For example, the pump assemblies provided in this embodiment may include P7 and P8. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the pump assemblies may also employ other configuration methods, locations, and quantities.

[0159] It is understood that, in addition, the magnetic bead purification process module 400 may also include several valve assemblies configured in various pipelines to control the on / off state of the pipelines. Exemplarily, the valve assemblies provided in the embodiments of this application may include V34 to V40. It is understood that this does not constitute an undue limitation on this application; in other embodiments, the valve assemblies may also employ other configuration methods, locations, and quantities, for example, being configured in various components that provide and collect solutions.

[0160] It is understood that the pre-purification process module may include both the magnetic bead purification process module 400 and the chromatography process module 300, or it may include only one of them. When both the magnetic bead purification process module 400 and the chromatography process module 300 are included, their order of arrangement can be selected as needed, and the examples in this application embodiment do not constitute an undue limitation.

[0161] In some embodiments, the apparatus for preparing nucleic acid drugs in this application may further include at least one mounting bracket, which can be used to mount process modules, such as a magnetic bead purification process module 400.

[0162] Please see here. Figure 26The mounting bracket includes a frame 1001 and a plurality of first connectors 1002 and a plurality of second connectors 1003 disposed on the frame. The first connectors 1002 extend along a first direction, and the second connectors 1003 extend along a second direction, with the first and second directions alternately arranged. For example, the first direction is vertical, and the second direction is horizontal. Of course, in other embodiments, the first and second directions are not limited to this arrangement.

[0163] Here, because the first connector 1002 and the second connector 1003 are staggered, multiple staggered points are formed in the frame, which can be used to install various components, such as at least one of the magnetic bead purification container 420, valve assembly, pump assembly, etc. Therefore, after the process flow is completed, consumables such as tubing can be quickly replaced. Since the positions of frequently used components are fixed, the operation of replacing consumables such as tubing is very convenient and can be easily achieved by mechanized methods such as robotic arms.

[0164] In some embodiments, the bottom end of the first connector 1002 may also be provided with a hook structure 1004 for connecting the required components.

[0165] It is understood that the mounting bracket is not limited to the installation of the magnetic bead purification process module 400; it can also be used to install other process modules, and the principle is the same.

[0166] Please see Figure 1 and Figure 2 The equipment for preparing nucleic acid drugs may also include an in vitro transcription device, which is located upstream to provide the drug solution for in vitro transcription. The transcribed drug solution can then be supplied to the first purification process module 500, or it can be supplied first to the pre-purification process module and then to the first purification process module 500. That is to say, the in vitro transcription device can be located upstream of the pre-purification process module and the first purification process module 500.

[0167] This may include a transfer unit 100 and a mixing unit 200.

[0168] The transfer unit 100 can be used to transfer at least two reactants into a reactor dish 12 (reactor dish 12 is visible). Figure 7 As shown, the mixing unit 200 can be used to control the reactor dish 12 within a preset temperature range and to mix the reactants in the reactor dish 12 to achieve in vitro transcription.

[0169] Therefore, by setting up the transfer unit 100 and the mixing unit 200, in vitro transcription can be effectively realized, which is more efficient than the existing technology of manually adding various reactants to the stirred tank.

[0170] The reactants are typically liquid reagents, which can be stored in a reactant container before in vitro transcription. The reactant container can be a test tube, other glass bottle, or other container, such as a vial. The quantity and type of reactants added will vary depending on the actual process requirements. The subsequent exemplary descriptions in this application do not constitute undue limitation, and those skilled in the art can adjust the quantity and type of reactants according to actual needs.

[0171] Please see here. Figure 1 To facilitate the arrangement of various components, a working platform 11 can be provided, on which each component can be mounted. For example, the transfer unit 100 and the mixing unit 200 can both be mounted on the working platform 11. Of course, in other embodiments, the working platform 11 may not be provided, and the examples in this application do not constitute an undue limitation.

[0172] For example, the in vitro transcription process module may also include a cooling chamber 160, a waste liquid chamber 170, and a buffer container 180. The cooling chamber 160 is used to refrigerate the reactants, as some reactants need to be stored within a suitable temperature range. The waste liquid chamber 170 is used to collect the waste liquid generated during the process operation. The waste liquid outlet of the in vitro transcription process module or other process modules can be connected to the waste liquid chamber 170. The buffer container 180 can be used to store the qualified drug solution after the in vitro transcription process is completed, so that it can be subsequently supplied to other process modules.

[0173] As another example, at least a portion of the refrigeration chamber 160 and the waste liquid chamber 170 are located below the work platform 11 to fully utilize the space below the work platform, improve the overall compactness of the process module, and reduce the occupied area. The buffer container 180 is disposed above the work platform for easy access and provision to other process modules.

[0174] In addition, the work platform 11 is also equipped with a refrigerated preparation storage area 191, a room temperature preparation storage area 192, and a pipette tip storage area 193. The refrigerated preparation storage area 191 can be located at the aforementioned refrigeration chamber 160. As another example, the refrigerated preparation storage area 191, the room temperature preparation storage area 192, and the pipette tip storage area 193 are all located on the same side of the transfer unit 100. Furthermore, the refrigerated preparation storage area 191, the room temperature preparation storage area 192, the pipette tip storage area 193, and the waste liquid chamber 170 are all located on the same side of the transfer unit 100 to facilitate the transfer unit 100 in taking and disposing of waste.

[0175] In addition, a laminar flow device can be installed above the work platform 11 so that each device can be located in a laminar flow environment, thereby achieving sterile isolation.

[0176] Please see here. Figure 3 The transfer unit 100 includes a first transfer member 120 and a first transfer driving mechanism 110 connected in a mating manner. The first transfer driving mechanism 110 is used to drive the first transfer member 120 to move so as to transfer at least two reactants into the reactor dish 12.

[0177] For example, the first transfer drive mechanism 110 may include a first lateral movement mechanism 111, a first rotation mechanism 112, and a first vertical movement mechanism 113. The first rotation mechanism 112 is disposed on the first lateral movement mechanism 111, thereby driving the first rotation mechanism 112 to move laterally. The first vertical movement mechanism 113 is disposed on the first rotation mechanism 112, thereby driving the first vertical movement mechanism 113 to rotate. The first vertical movement mechanism 113 is connected to the first transfer member 120, thereby driving the first transfer member 120 to move vertically up and down.

[0178] Furthermore, through the provision of the first transfer drive mechanism 110, the first transfer member 120 can drive the first transfer drive mechanism 110, thereby realizing lateral movement, rotation in the horizontal plane, and vertical movement.

[0179] For example, the first lateral moving mechanism 111 may include a horizontally arranged linear module; the first rotating mechanism 112 may be a motor, which is directly or indirectly connected to the slider of the first lateral moving mechanism 111 through a connector; the first vertical moving mechanism 113 may include a vertically arranged linear module, which is directly or indirectly connected to the first rotating mechanism 112 through a connector; and the first picking member 120 is connected to the slider of the first vertical moving mechanism 113.

[0180] Of course, it is understood that the structure of the first transfer drive mechanism 110 and the structure and connection positions of its components are not limited to the above examples, as long as they can drive the first transfer member 120 to move to the desired position.

[0181] Exemplarily, the first transfer member 120 may include a pipette tip 121 and a pressure source 122, the pressure source 122 being connected to the pipette tip 121 to create a negative pressure for adsorbing the reactants. For example, the pressure source 122 may be a pump or the like. By changing the pressure of the pressure source 122, the adsorption and release of the reactants can be achieved.

[0182] For example, the first transfer member 120 also includes a pipette tip mounting portion 123, which can be detachably connected to the pipette tip 121 by means of plugging or the like. In addition, the pipette tip mounting portion 123 may also be provided with an air passage so that the pipette tip 121 mounted on the pipette tip mounting portion 123 can be connected to the pressure source 122.

[0183] For example, the pipette tip 121 can be a disposable tip, and different pipette tips 121 can be used when transferring different reactants.

[0184] In some examples, a rotary encoder may also be provided at the first rotating mechanism 112. The rotary encoder can obtain rotational parameters, which may include the angular displacement, angular velocity, etc. of the output shaft of the first rotating mechanism 112. Furthermore, a pipette tip placement area is pre-set, with several pipette tips 121 arranged therefor connecting with the tip mounting part 123. Position parameters of the several pipette tips 121 are obtained through pre-positioning. By calculating the position of the tip mounting part 123, the position of the first lateral moving mechanism 111, and the position parameters of the pipette tips 121, the rotational parameters required to move to the position of each pipette tip 121 can be obtained. Therefore, by controlling the rotational parameters of the rotary encoder, the tip mounting part 123 can accurately move to the position of the pipette tip 121 and connect with it.

[0185] In addition, in some examples, the aforementioned pressure source 122 is also used to intermittently create negative pressure, thereby repeatedly drawing in and releasing reactants by changing between negative and positive pressure, that is, stirring the reactants by changing the air pressure, thereby mixing the reactants.

[0186] In some embodiments, please continue reading Figure 3 The aforementioned transfer unit 100 may further include a second transfer member 140 and a second transfer drive mechanism 130 that are connected in a mating manner. The second transfer drive mechanism 130 is used to drive the second transfer member 140 to move in order to transfer the reactant container. Thus, the reactant container can be transferred to the desired location.

[0187] Furthermore, by way of example, the reactant container may also include a lid (not shown), and the second transfer member 140 may also be used to transfer the lid, thereby enabling the lid opening operation. That is, the second transfer member 140 may be used to transfer at least one of the reactant container and the lid.

[0188] The connection between the cover and the reactant container can be a threaded fit or an interference fit, etc. It is understood that the example in this embodiment does not constitute an undue limitation.

[0189] As another example, the second transfer drive mechanism 130 may include a second lateral movement mechanism 131, a second rotation mechanism 132, and a second vertical movement mechanism 133. The second rotation mechanism 132 is disposed on the second lateral movement mechanism 131, thereby allowing the second lateral movement mechanism to drive the second rotation mechanism 132 to move laterally. The second vertical movement mechanism 133 is disposed on the second rotation mechanism 132, thereby allowing the second rotation mechanism 132 to drive the second vertical movement mechanism 133 to rotate. The second vertical movement mechanism 133 is connected to the second transfer member 140, thereby allowing the second transfer member 140 to move vertically up and down.

[0190] Furthermore, through the provision of the second transfer drive mechanism 130, the second transfer member 140 can drive the second transfer drive mechanism 130, thereby realizing lateral movement, rotation in the horizontal plane, and vertical movement.

[0191] For example, the second lateral moving mechanism 131 may include a horizontally arranged linear module; the second rotating mechanism 132 may be a motor, which is directly or indirectly connected to the slider of the second lateral moving mechanism 131 through a connector; the second vertical moving mechanism 133 may include a vertically arranged linear module, which is directly or indirectly connected to the second rotating mechanism 132 through a connector; and the second picking member 140 is connected to the slider of the second vertical moving mechanism 133.

[0192] Of course, it is understood that the structure of the second transfer drive mechanism 130 and the structure and connection positions of its components are not limited to the above examples, as long as they can drive the second transfer member 140 to move to the desired position.

[0193] For example, the second transfer member 140 may be a structure such as an electric or pneumatic gripper or a robotic arm.

[0194] In some embodiments, please refer to Figure 2 The aforementioned transfer unit 100 may further include a clamping mechanism 150 for clamping and securing the reactant container. For example, the second transfer member 140 can transfer the reactant container to the position of the clamping mechanism 150, and the clamping mechanism 150 clamps and secures the reactant container.

[0195] Here, for example, please refer to Figure 4 The clamping mechanism 150 includes a clamping member 151 and a clamping drive mechanism (not shown) that are connected to each other. The clamping member 151 can be a gripper or the like, and the clamping drive mechanism can be a motor or the like. When the second transfer member 140 transfers the reactant container to the clamping member 151, the clamping drive mechanism drives the clamping member 151 to move in order to clamp and fix the reactant container.

[0196] Here, multiple clamping mechanisms 150 can be provided as needed, for example in... Figure 4 In the schematic diagram shown, two clamping mechanisms 150 are arranged side by side. Of course, the examples in this application embodiment do not constitute an undue limitation. In other embodiments, the clamping mechanisms 150 may be configured in other numbers, and their arrangement positions may also be different.

[0197] Please see Figure 5 The aforementioned mixing unit 200 may include a carrier 210, a mixing drive mechanism 220, and a temperature control mechanism (not shown).

[0198] The carrier 210 is used to detachably fix the reactor dish 12. The mixing drive mechanism 220 is connected to the carrier 210 to drive the carrier 210 to reciprocate. The temperature control mechanism is directly or indirectly connected to the reactor dish 12 to transfer heat to the reactor dish 12.

[0199] Thus, at the mixing unit 200, the reactants can be controlled within a preset temperature range by the temperature control mechanism and driven by the mixing drive mechanism 220 to achieve the enzyme reaction and thus achieve in vitro transcription.

[0200] Please refer to the following for details. Figure 6 The mixing drive mechanism 220 includes a power source 221 and an eccentric transmission structure connected in conjunction, with the carrier 210 connected to the eccentric transmission structure. Thus, driven by the power source 221, the carrier 210 can rotate via the transmission of the eccentric transmission structure. Since the reactor dish 12 is detachably fixed to the carrier 210, it can further rotate, thereby achieving uniform mixing of the reactants within the reactor dish 12.

[0201] For example, the power source 221 can be a motor or the like. Also for example, the eccentric transmission mechanism includes a drive eccentric wheel 222, which is driveably connected to the output shaft of the aforementioned power source 221. An eccentrically configured first eccentric transmission member 223 is connected to the drive eccentric wheel 222. Specifically, the first eccentric transmission member 223 may be non-coaxial with the output shaft of the power source 221. The first eccentric transmission member 223 is used to connect the carrier member 210. In some examples, the end of the first eccentric transmission member 223 away from the drive eccentric wheel 222 may extend radially to form a bearing surface for connecting the carrier member 210, thereby improving the stability of the connection.

[0202] In a further embodiment, the eccentric transmission mechanism may also include a passive eccentric wheel 224, which is rotatably mounted on a base 226. An eccentrically positioned second eccentric transmission member 225 is connected to the passive eccentric wheel 224. Specifically, the second eccentric transmission member 225 may be non-coaxial with the center of the passive eccentric wheel 224. The second eccentric transmission member 225 also connects to the carrier member 210 to improve the stability of the carrier member 210 during movement. In some examples, the second eccentric transmission member 225 may extend radially at one end near the carrier member 210 to form a bearing surface for connecting to the carrier member 210, thereby improving the stability of the connection. Furthermore, the area of ​​the bearing surface of the second eccentric transmission member 225 may be larger than the area of ​​the bearing surface of the first eccentric transmission member 223.

[0203] In some examples, multiple passive eccentric wheels 224 are configured, arranged circumferentially around the active eccentric wheel, for example, they can be evenly spaced. For example, in... Figure 6 In this embodiment, three passive eccentric wheels 224 are evenly spaced around one active eccentric wheel in the circumference. Of course, the example in this embodiment does not constitute an undue limitation.

[0204] Please refer to the following: Figure 5 The carrier 210 may include a heat-insulating container 212, which may be disposed on the aforementioned mixing drive mechanism 220 and driven by the mixing drive mechanism 220 to reciprocate. For example, the heat-insulating container 212 may be directly or indirectly disposed on the first eccentric transmission member 223 and the second eccentric transmission member 225. Exemplarily, the carrier 210 includes a heat insulation seat 211, a heat-insulating container 212, and a heat-insulating cover 213. The heat insulation seat 211 is disposed on the first eccentric transmission member 223 and the second eccentric transmission member 225. The heat-insulating container 212 is assembled inside the heat insulation seat 211, and the heat-insulating container 212 has an openable heat-insulating cover 213.

[0205] Please refer to the following for details. Figure 7 The illustration shows a reactor dish 12 placed in an insulated container 212, with the insulated lid 213 opened and removed. The reactor dish 12 is exemplarily a conical flask. The insulated lid 213 can be removed via the aforementioned second removal member 140 to achieve opening and removal.

[0206] For another example, please combine Figure 8 And mainly refer to Figure 9 The insulated container 212 may be provided with at least two elastic elements 214, such as spring screws, and the elastic elements 214 may be spaced apart circumferentially. For example, three elastic elements 214 are provided circumferentially here.

[0207] Therefore, when the reactor dish 12 is located inside the insulated container 212, the elastic element 214 abuts against the insulated container 212 and the reactor dish 12. When the mixing drive mechanism 220 drives the insulated container 212 and the reactor dish 12 to move, the elastic element 214 helps to achieve a buffering and positioning function, preventing the inner wall of the insulated container 212 and the outer wall of the reactor dish 12 from colliding with each other. At the same time, the elastic element 214 also plays a positioning role for the reactor dish 12, so as to facilitate the gripping and removal of the reactor dish 12.

[0208] The temperature control mechanism may include a heating component (not shown in the figure), which may be located inside the insulated container 212 or at the bottom of the insulated container 212, etc., to heat the reactor dish 12 inside the insulated container 212.

[0209] In addition, in some embodiments, please refer to [further details]. Figure 5 The mixing unit 200 may also include a tilting component 240, which is connected to one side of the support member 210 so as to drive one side of the support member 210 to lift it, thereby tilting the support member 210 and the reactor vessel 12 thereon, which makes it easier to remove the solution that has completed the reaction.

[0210] For example, the tilting assembly 240 may include a tilting motor 241, which may be directly or indirectly connected to a connecting frame 243. The aforementioned hybrid drive mechanism 220 and the support member 210 may be directly or indirectly mounted on the connecting frame 243. Thus, when the tilting motor 241 is started, the hybrid drive mechanism 220, the support member 210, etc., can be tilted. Here, a reducer 242 may be driven between the tilting motor 241 and the support member 210.

[0211] In some embodiments, please refer to Figure 5 The mixing unit 200 may also include a positioning component 244 for gripping and fixing the reactor dish 12 so that the second transfer member 140 can remove the lid of the reactor dish 12. The positioning component 244 may be an electric gripper, a pneumatic gripper, or the like.

[0212] When the second transfer member 140 removes the lid of the reactor dish 12, the positioning component 244 can clamp the neck of the reactor dish 12 to achieve fixation, thereby facilitating the removal of the lid of the reactor dish 12 by the second transfer member 140.

[0213] In some embodiments, please refer to Figure 10The buffer container 180 is also provided. For example, the buffer container 180 can be connected to the aforementioned reactor dish 12 via a pipette assembly. For instance, after the tilting assembly 240 tilts the reactor dish 12, the pipette tip is driven by a motor, a robotic arm, or other mechanism and moves into the reactor dish 12, thereby drawing the liquid medicine into the buffer container 180 through the pipette assembly's tubing and through components such as a pump configured in the tubing.

[0214] In some embodiments, the buffer container 180 is mounted on a base 181. See also Figure 11 A magnetic rotor 1801 is installed at the bottom of the buffer container 180. Please combine this with... Figure 12 The base 181 is provided with a magnetic drive component 1811, which is used to generate a magnetic field driving force to drive the magnetic rotor 1801 to rotate, thereby achieving the mixing of the liquid medicine.

[0215] The base 181 may be provided with a first positioning structure 1812, and the buffer container 180 may be provided with a second positioning structure 1802, so that the two can be quickly engaged and disassembled. For example, one of the first positioning structure 1812 and the second positioning structure 1802 is a protrusion and the other is a groove, and the two can be interference-fitted to achieve snap-fit, etc.

[0216] In some embodiments, please refer to Figure 10 A weighing component 1813 may also be provided on the base 181 to weigh the liquid medicine in the buffer container 180.

[0217] In some embodiments, to better achieve in vitro transcription, a premixing unit may be provided upstream of the mixing unit 200 to better mix the reactants before the reaction. Furthermore, in some embodiments, the premixing component may also be used to perform IVT and co-transcriptional capping reactions. In some embodiments, reactants may be further added and mixed to complete the tailing reaction. After these reactions are completed, an enzyme termination reaction is performed.

[0218] The premixing unit may include a premixing container and a premixing assembly. The premixing assembly may include a premixing carrier and a premixing drive mechanism. The premixing drive mechanism is connected to the premixing carrier to drive the premixing carrier to reciprocate, thereby achieving mixing. For example, the premixing drive mechanism includes a premixing motor, which is connected to the premixing carrier via an eccentric transmission component to drive the premixing carrier to move. The premixing carrier may be a platform or a gripper, etc.

[0219] In some examples, the structure of the premixed component may also be set to be the same as or similar to the structure of the aforementioned mixing drive mechanism 220.

[0220] In practical use, the reactants are loaded into reactant containers before being removed by the first transfer member 120. The reactant container of any reactant can be used as a premixing container. Thus, the reactant container can be used as a premixing container, thereby reducing consumables and also reducing the number of process steps for transferring reactants.

[0221] In some embodiments, the carrier 210 reciprocates along a first trajectory, and the premixed carrier reciprocates along a second trajectory, the first trajectory being different from the second trajectory. For example, the first trajectory is set along a horizontal plane, and the second trajectory is set along a vertical plane. By performing mixing operations in different directions, a better mixing effect can be achieved.

[0222] In some embodiments, the reciprocating frequency of the carrier 210 is a first frequency, and the reciprocating frequency of the premixing carrier is a second frequency. The first frequency is higher than the second frequency, thereby enabling better premixing in the early stage and meeting the reaction requirements of the enzyme reaction in the later stage through the reciprocating movement of the second frequency.

[0223] Accordingly, this application also provides a method for preparing nucleic acid drugs, which can use the equipment for preparing nucleic acid drugs provided in any of the foregoing embodiments.

[0224] Please see Figure 27 The method includes the following steps S400 to S600.

[0225] In step S400, the input solution is purified by the first purification process module 500.

[0226] For example, step S400 includes a purification step, which includes the following steps.

[0227] The solution is passed through the first separation and purification device 520 multiple times via the first circulation loop.

[0228] Displacement solution is added to the first mixing container 525 through the first displacement solution inlet 526, and the displacement solution and the solution are circulated together in the first circulation loop and flow through the first separation and purification device 520 multiple times.

[0229] Following the purification step, a solution output step is also provided, which includes the following steps.

[0230] Replacement fluid is added to the first separation and purification device 520 through the first replacement fluid inlet 526, so as to push the purified solution out of the first separation and purification device 520.

[0231] The solution flows to the first filtration component 531 and, after filtration, flows to the first purification collection end 530.

[0232] A post-processing step is provided after the solution output step, which includes the following steps.

[0233] Gas is input through the first gas inlet 550, and after flowing through the first filter assembly 531, it flows to the second waste liquid collection end 541.

[0234] When gas flows through the first filter component 531, the first detection sensor 551 detects whether the first filter component 531 is damaged.

[0235] In step S500, lipid nanoparticles are prepared using a lipid nanoparticle preparation process module based on the purified solution.

[0236] The method for preparing lipid nanoparticles is existing technology, and will not be described in detail in the embodiments of this application.

[0237] In step S600, the solution containing lipid nanoparticles is purified by the second purification process module 700.

[0238] Step S600 includes a pre-preparation step, which includes the following steps.

[0239] Pure water is input through the second pre-preparation inlet 740, and after flowing through the second separation and purification device 720, it flows to the third waste liquid collection end 724.

[0240] Alkali solution is introduced through the second pre-preparation inlet 740. After flowing through the second separation and purification device 720, the alkali solution flows to the third waste liquid collection end 724.

[0241] Pure water is reintroduced through the second pre-preparation inlet 740, and after flowing through the second separation and purification device 720, it flows to the third waste liquid collection end 724.

[0242] The rinsing is performed by introducing equilibration fluid through the second pre-preparation inlet 740.

[0243] In some examples, the other steps after the pre-preparation step are similar to step S400. For details, please refer to the descriptions in other embodiments of this application specification. To avoid making the application document too lengthy, they will not be described in detail here.

[0244] In some embodiments, before purifying the solution by the first purification process module 500, a magnetic bead purification operation is also performed by the magnetic bead purification process module 400, which includes the following steps.

[0245] The solution is introduced into the magnetic bead purification circulation loop through the magnetic bead purification inlet 410 and circulated several times.

[0246] The magnetic beads are adsorbed by the magnetic source 460, and the magnetic beads are retained in the magnetic bead purification container 420. The solution is discharged to the waste liquid outlet 440.

[0247] Washing liquid is introduced through the magnetic bead purification inlet 410. The magnetic source 460 is removed, so that the magnetic beads are released from fixation and come into contact with the washing liquid. At the same time, the washing liquid and the magnetic beads are vibrated by the oscillation mechanism. After washing for a first preset time, the magnetic source 460 is moved back and fixes the magnetic beads. Then the washing liquid is discharged to the waste liquid outlet 440.

[0248] Eluent is introduced through the magnetic bead purification inlet 410. The magnetic source 460 is removed, which releases the magnetic beads from fixation and brings them into contact with the eluent. The oscillation mechanism causes the eluent and the magnetic beads to vibrate. After elution for a second preset time, the magnetic source 460 is moved back and fixes the magnetic beads. The eluent is then transported to other downstream components through the magnetic bead purification outlet 450.

[0249] The first and second preset times are set according to process requirements and actual conditions. They can be the same or different. The examples in this application do not constitute an undue limitation on them.

[0250] In some embodiments, before purifying the solution by the first purification process module 500, a chromatography operation is performed by the chromatography process module 300, the chromatography operation including the following steps.

[0251] The solution is fed into the chromatography column.

[0252] The chromatography column is subjected to at least two gradient elutions using an elution solution of different concentrations for each gradient elution.

[0253] The chromatographic solution is obtained from the chromatography column.

[0254] In some embodiments, before performing magnetic bead purification, chromatography, or a first purification operation, a solution after in vitro transcription is obtained using an in vitro transcription device, and the above-described operations are performed on the solution.

[0255] In some embodiments, the preparation method of the nucleic acid drug solution further includes an in vitro transcription method, which can employ the in vitro transcription process module provided in any of the foregoing embodiments. The in vitro transcription method is used to prepare the drug solution for subsequent first purification and / or pre-purification operations.

[0256] Here, the in vitro transcription method includes the following steps S100 to S200.

[0257] In S100, at least two reactants are transferred into the reactor vessel 12 by the transfer unit 100.

[0258] Here, in some embodiments, prior to S100, at least two reactants may be premixed using a premixing component. Exemplarily, during premixing, the reactant container of any reactant is used as the premixing container.

[0259] Here, in some embodiments, when any of the reactants is removed by the transfer unit 100, a negative pressure is intermittently formed to mix the reactants.

[0260] In S200, the mixing unit 200 drives the reactor dish 12 to move back and forth and controls the temperature of the reactor dish 12 within a preset temperature range to achieve the enzyme reaction.

[0261] For example, the mixing unit 200 controls the temperature of the reactor dish 12 at 37 degrees Celsius and incubates the reactants inside for 3 hours. Afterwards, the transfer unit 100 injects other solutions into the reactor dish 12 again, and the reactor dish 12 undergoes another isothermal oscillation operation. It is understood that the specific temperature and duration of the isothermal oscillation are related to the specific process and can be selected differently in different embodiments. The examples in this application do not constitute an undue limitation.

[0262] In some embodiments, after the enzyme reaction is completed, one side of the carrier 210 is lifted by the tilting component 240 to facilitate the removal of the solution after the reaction is completed.

[0263] In the in vitro transcription process module and method provided in this application embodiment, the in vitro transcription process module includes a first transfer element 120 and a first transfer driving mechanism 110 connected in cooperation. The first transfer driving mechanism 110 drives the first transfer element 120 to move, thereby transferring at least two reactants into the reactor dish 12. Then, the mixing unit 200 drives the reactor dish 12 to reciprocate, and the temperature control mechanism of the mixing unit 200 transfers heat to the reactor dish 12 to realize the enzyme reaction. Thus, a highly efficient in vitro transcription operation is achieved.

[0264] In addition, premixing can be performed at least once before the enzyme reaction, thereby improving the effectiveness and efficiency of in vitro transcription.

[0265] Furthermore, the efficiency of in vitro transcription operations can be further improved through the cooperation of the second transfer element 140 and the second transfer drive mechanism 130.

[0266] After completing the in vitro transcription operation, the solution was purified and prepared into lipid nanoparticles through the first purification operation, lipid nanoparticle preparation, and second purification operation. The two purification operations improved the effective components of the final nucleic acid drug.

[0267] Furthermore, the relevant operations can be carried out using automated equipment for preparing nucleic acid drugs, eliminating the need for manual operation of the entire process and thus improving preparation efficiency. Example

[0268] Here, the in vitro transcription operation in the preparation method of nucleic acid drugs is explained using the example of two reactants. It is understood that unless otherwise specified, the order of the steps described in this application does not represent a restriction on their order.

[0269] In specific operation, the second transfer member 140 transfers the two reactant containers to the clamping mechanism 150, which clamps and secures the two reactant containers. Furthermore, the second transfer member 140 can also be used to remove the lids of the reactant containers to open them.

[0270] The first transfer element 120 moves to the position of one of the reactant containers and repeatedly draws in and releases the reactants in the reactant container by intermittently creating negative pressure, thereby achieving mixing.

[0271] After mixing, the first transfer member 120 transfers the reactant to the reactant container of another reactant.

[0272] The second transfer member 140 transfers a reactant container containing two reactants to a premixing unit, which shakes the reactant container to achieve premixing.

[0273] Exemplarily, the aforementioned mixing and / or premixing steps can achieve the mixing of at least two reactants, thereby enabling the IVT reaction and co-transcriptional capping reaction. In some embodiments, reactants may be further added and mixed to complete the tailing reaction. After these reactions are completed, an enzyme termination reaction is performed.

[0274] After premixing, the first transfer device 120 transfers the solution from the reactant container to the reactor dish 12, and the second transfer device 140 closes the insulated lid 213 and / or the lid of the reactor dish 12. Here, the drug solution is incubated and the enzyme-terminating reaction is initiated, for example, by incubating at 37°C for 3 hours.

[0275] The reactor vessel 12 is placed on the support 210. The temperature control mechanism heats the reactor vessel to a preset temperature range, and the mixing drive mechanism 220 drives the reactor vessel 12 to reciprocate to realize the enzyme reaction.

[0276] After the enzyme reaction is completed, the second transfer member 140 opens the heat preservation cover 213 and / or the lid of the reactor dish 12. When opening the lid of the reactor dish 12, the reactor dish 12 can be clamped by the positioning component 244 so that the second transfer member 140 can remove the lid of the reactor dish 12.

[0277] After the lid is removed, the tilting component 240 tilts the reactor vessel 12 to allow the solution to be removed. Example

[0278] When more reactants are required according to process requirements, more reactants can be added through the second transfer member 140 according to process requirements.

[0279] For example, the three reactants can be transferred into the reactant container of any one of the reactants via the second transfer member 140, after premixing.

[0280] For example, before or after the first transfer member 120 transfers the solution from the premixed reactant container to the reactor vessel 12, other reactants can be added to the reactor vessel 12 through the first transfer member 120. Example

[0281] Before performing the first purification operation, a preparatory step is first taken.

[0282] In the pre-preparation step, pure water can be input through the first pre-preparation inlet 540. After the pure water flows through the first separation and purification device, it flows to the second waste liquid collection end 541, thereby achieving the cleaning of hollow fibers, etc.

[0283] Then, alkaline solution is introduced through the first pre-preparation inlet 540. After flowing through the first separation and purification device, the alkaline solution flows to the second waste liquid collection end 541. Then, pure water can be introduced again for cleaning, and the flow path is the same.

[0284] Then, the equilibration solution can be introduced through the first pre-preparation inlet 540 for rinsing. After flowing through the first separation and purification device, the equilibration solution is divided into two streams, flowing to the first waste liquid collection end 524 and the second waste liquid collection end 541 respectively.

[0285] During the rinsing process with the equilibration fluid, each of the first sensing components 542 in the pipeline can be calibrated. For example, the first sensing component 542 includes a pH sensor, which can be calibrated by comparing the value when the equilibration fluid flows through it with a reference value.

[0286] After the preparatory steps are completed, the purification step is carried out.

[0287] In the purification step, the solution from the upstream, such as the solution from the in vitro transcription device, can be circulated in the first loop and then flow through the first separation and purification device multiple times to achieve purification.

[0288] Then, the replacement liquid can be added to the first mixing container 525 through the first replacement liquid inlet 526, and the replacement liquid and the aforementioned solution can be circulated together in the first circulation loop, so that they can flow through the first separation and purification device multiple times to achieve purification.

[0289] After the purification step is completed, the solution output step is performed.

[0290] In the solution output step, replacement fluid can be added to the first separation and purification device through the first replacement fluid inlet 526 to fully push the purified solution out of the first separation and purification device. This step helps to reduce losses.

[0291] The replacement fluid has a preset flow rate, which is determined by prior calculation to prevent excessive addition of replacement fluid and its flow to the first purification collection end 530 and the first filter assembly 531.

[0292] Subsequently, the solution flows to the first filtration assembly 531 and, after filtration, flows to the first purification collection end 530.

[0293] After the solution output step is completed, a post-processing step can be performed.

[0294] In the post-processing step, pure water can be introduced through the first pre-preparation inlet 540. After the pure water flows through the first separation and purification device, it flows to the second waste liquid collection end 541, thereby achieving cleaning.

[0295] In addition, gas can be introduced through the first gas inlet 550, and after flowing through the first filter assembly, the gas flows to the second waste liquid collection end 541, which helps to drain the liquid in the pipeline. Furthermore, when the gas flows through the first filter assembly, a first detection sensor 551, such as a pressure sensor, can detect whether the first filter assembly is damaged.

[0296] After the first extraction operation is completed, the produced liquid is transported through pipelines to the downstream lipid nanoparticle preparation process module for the preparation of lipid nanoparticles.

[0297] After the preparation of lipid nanoparticles is completed, a second purification operation can be performed through the second purification process module 700.

[0298] Before proceeding with the second purification operation, a preparatory step is first performed.

[0299] In the pre-preparation step, pure water can be input through the second pre-preparation inlet 740. After the pure water flows through the second separation and purification device, it flows to the third waste liquid collection end 724, thereby achieving the cleaning of hollow fibers, etc.

[0300] Then, alkaline solution is introduced through the second pre-preparation inlet 740. After flowing through the second separation and purification device, the alkaline solution flows to the second waste liquid collection end 541. Then, pure water can be introduced again for cleaning, and the flow path is the same.

[0301] Then, the equilibration solution can be introduced through the second pre-preparation inlet 740 for rinsing. After flowing through the second separation and purification device, the equilibration solution is divided into two streams, flowing to the third waste liquid collection end 724 and the fourth waste liquid collection end 741 respectively.

[0302] During the rinsing process with the equilibration fluid, each of the second sensing components 742 in the pipeline can be calibrated. For example, the second sensing component 742 includes a pH sensor, which can be calibrated by comparing the value when the equilibration fluid flows through it with a reference value.

[0303] After the preparatory steps are completed, the purification step is carried out.

[0304] In the purification step, the solution from the upstream, such as the solution from the lipid nanoparticle preparation process module, can be circulated in the second circulation loop and then flow through the second separation and purification device multiple times to achieve purification.

[0305] Then, replacement liquid, mother liquor, auxiliary material mother liquor, etc. can be added to the second mixing container 725 through the second replacement liquid inlet, and the replacement liquid and the aforementioned solutions can be circulated together in the second circulation loop, so that they can flow through the second separation and purification device multiple times to achieve purification.

[0306] After the purification step is completed, the solution output step is carried out.

[0307] In the solution output step, replacement fluid can be added to the second separation and purification device through the second replacement fluid inlet to fully push the purified solution out of and / or close to the second separation and purification device. This step helps to reduce losses.

[0308] The replacement fluid has a preset flow rate, which is determined by prior calculation to prevent excessive addition of replacement fluid and its flow into the second purification and collection container 730 and the second filter assembly 731.

[0309] Subsequently, the solution flows to the second filtration assembly 731 and, after filtration, flows to the second purification collection container 730.

[0310] After the solution output step is completed, a post-processing step can be performed.

[0311] In the post-processing step, pure water can be introduced through the second pre-preparation inlet 740. After flowing through the second separation and purification device, the pure water flows to the fourth waste liquid collection end 741, thereby achieving cleaning.

[0312] In addition, gas can be introduced through the second gas inlet 750. After flowing through the second filter, the gas flows to the fourth waste liquid collection end 741, which helps to drain the liquid in the pipeline. Furthermore, when the gas flows through the second filter, a second detection sensor 751, such as a pressure sensor, can be used to detect whether the second filter is damaged. Example

[0313] Before performing the first purification operation, magnetic bead purification can be performed using the magnetic bead purification process module 400.

[0314] The solution, which originates from upstream sources such as in vitro transcription equipment, contains magnetic beads. These beads are then introduced into the magnetic bead purification loop through the magnetic bead purification inlet 410 and circulated several times.

[0315] Then, the magnetic source 460 at the magnetic bead purification container 420 is activated. During the solution flow, the magnetic beads are adsorbed by the magnetic source 460, thereby separating the magnetic beads from the solution. The magnetic beads are retained in the magnetic bead purification container 420, and the solution is discharged to the waste liquid outlet 440.

[0316] Then, washing liquid is introduced through the magnetic bead purification inlet 410. During this process, the magnetic source 460 is removed, allowing the magnetic beads to be released from fixation and come into contact with the washing liquid. At the same time, the oscillation mechanism causes the washing liquid and the magnetic beads to vibrate for better contact. After washing is completed, the magnetic source 460 is moved back to fix the magnetic beads, and then the washing liquid is discharged to the waste liquid outlet 440.

[0317] Subsequently, eluent is introduced through the magnetic bead purification inlet 410. During this process, the magnetic source 460 is removed, allowing the magnetic beads to be released from fixation and come into contact with the eluent. Simultaneously, an oscillation mechanism promotes vibration between the eluent and the magnetic beads for better contact. After elution is complete, the magnetic source 460 is moved back to fix the magnetic beads, and the eluent is then transported through the magnetic bead purification outlet 450 to other downstream components, such as the first purification process module. Example

[0318] Before performing the first purification operation, a chromatography operation can be performed using the chromatography process module 300.

[0319] During the chromatography operation, a calibration solution is first introduced through the chromatography inlet line 310. The calibration solution flows from the first port 321 to the fourth port 324, bypassing the chromatography column, and then enters the chromatography outlet line 330. During this process, the readings of various sensing components 350 in the lines are calibrated to confirm that the readings of the sensing components 350 are consistent with the parameters that the calibration solution should have, thereby confirming the status of the sensing components 350.

[0320] Then, the equilibrium solution and other solutions are introduced through the chromatography inlet pipe 310. The solution flows to the chromatography column through the first port 321 and the second port 322, and then enters the chromatography outlet pipe 330 through the third port 323 and the fourth port 324.

[0321] The above steps are the preparation steps for the chromatography operation. If the chromatography process module 300 has completed the relevant calibration, column equilibration and other operations in advance through other means, the above steps can be omitted.

[0322] A solution from upstream, such as that from an in vitro transcription device, is introduced into the chromatography inlet line 310. The solution flows through the first port 321 and the second port 322 to the chromatography column, and then through the third port 323 and the fourth port 324 into the chromatography outlet line 330. At this point, the desired substance is retained in the chromatography column, while other solutions are discharged.

[0323] Then, the eluent is introduced into the chromatography column through the chromatography inlet line 310 for elution. The eluent can be an alkaline solution to remove any remaining solutions from the chromatography column. In some examples, elution can be performed more than twice, each time using an eluent of a different concentration to achieve gradient elution.

[0324] Then, the eluent is introduced through the chromatography inlet line 310. The eluent flows to the chromatography column through the first port 321 and the second port 322, and then enters the chromatography outlet line 330 through the third port 323 and the fourth port 324. This eluent removes the desired substance from the chromatography column and dissolves it in the eluent. This eluent containing the dissolved substance is then introduced into downstream components, such as the first purification process module 500, through the chromatography outlet line 330.

[0325] It is understood that, in the embodiments described in this application, in order to illustrate their function, the names of some of the required solutions in steps such as chromatography, magnetic bead purification, first purification, and second purification may be similar or the same, such as eluent. However, in different components, the specific solutions used in the actual process will be determined according to the specific process requirements, and it does not mean that the composition of the solutions used in different components is the same or different.

[0326] It is understood that the terms used in the embodiments of this application have the same meaning. For any content not described in detail in a certain embodiment, the specific implementation details can be referred to the descriptions in other embodiments. The examples and technical effects shown in the foregoing embodiments can be implemented accordingly. For repeated parts, this embodiment will not elaborate further.

[0327] The above provides a detailed description of the equipment and method for preparing nucleic acid drugs provided in this application. Specific examples have been used to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the methods and core ideas of this application. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the ideas of this application. Therefore, the content of this specification should not be construed as a limitation of this application.

Claims

1. A purification apparatus for preparing nucleic acid drugs, characterized in that, It includes a first purification process module, which includes, The first separation and purification device has a first inlet end, a first outlet end and a first waste liquid outlet end. The first separation and purification device is configured in a first circulation loop, and the first circulation loop has a first purification input end. A first mixing container is configured in the first circulation loop, with its inlet connected to the first outlet and its outlet connected to the first inlet. The first displacement fluid inlet is connected to the inlet of the first mixing container; and Pump assembly used to drive the flow of solution.

2. The purification apparatus for preparing nucleic acid drugs according to claim 1, characterized in that, The first purification process module also includes, The first purification and collection end is connected to the first mixing container; as well as A first filtration component is disposed between the first mixing container and the first purification collection end.

3. The purification apparatus for preparing nucleic acid drugs according to claim 2, characterized in that, The first purification process module also includes, The first gas inlet is connected to the first filter assembly; as well as The first detection sensor is used to detect pressure changes in the pipeline where the first filter assembly is located when gas is input into the first filter assembly at the first gas inlet end.

4. The purification apparatus for preparing nucleic acid drugs according to claim 1, characterized in that, The first purification process module also includes, At least one first pre-prepared inlet end; And, the second waste liquid collection end; The first separation and purification device is configured via a pipeline between the first pre-preparation inlet and the second waste liquid collection end.

5. The purification apparatus for preparing nucleic acid drugs according to claim 1, characterized in that, The first mixing container includes, Container body; The inlet pipe extends into the container body and then extends toward the side wall of the container body; An outlet pipe extends to the bottom of the container body; as well as The stirring component includes a stirring drive mechanism and a stirring part that are connected in a mating manner, with the stirring part located within the container body.

6. The purification apparatus for preparing nucleic acid drugs according to claim 1, characterized in that, The first mixing container is disposed on a mixing mechanism, the mixing mechanism comprising: Mixing drive component; An eccentric connector is eccentrically connected to the mixing drive. The support member is rotatably connected to the eccentric connector; When the mixing drive unit drives the eccentric connector to move, it can cause the support and the first mixing container to rotate and shake, so as to mix the medicine liquid in the first mixing container.

7. The purification apparatus for preparing nucleic acid drugs according to claim 1, characterized in that, It also includes a second purification process module, located downstream of the first purification process module. The second purification process module includes... The second separation and purification device has a second inlet end, a second outlet end and a second waste liquid outlet end. The second separation and purification device is configured in a second circulation loop. The second circulation loop has a second purification input end. The second mixing container is configured in the second circulation loop, and its inlet is connected to the second outlet end and its outlet is connected to the second inlet end; At least two auxiliary liquid inlet terminals are connected to the inlet of the first mixing container; as well as Pump assembly used to drive the flow of solution.

8. The purification apparatus for preparing nucleic acid drugs according to claim 7, characterized in that, The second purification process module also includes, The second purification and collection end is connected to the second mixing container; as well as The second filtration device is disposed between the second mixing container and the second purification collection end.

9. The purification apparatus for preparing nucleic acid drugs according to claim 1, characterized in that, At least one pre-purification process module is also provided upstream of the first purification process module.

10. The purification apparatus for preparing nucleic acid drugs according to claim 9, characterized in that, The pre-purification process module includes a chromatography process module, which includes, The chromatography main pipeline includes a first port and a second port for fluid connection, as well as a third port and a fourth port for fluid connection; The chromatography inlet pipe is connected to the first port; A chromatography column, the inlet of which is connected to the second port and the outlet of which is connected to the third port; The chromatography outlet pipe is connected to the fourth port.

11. The purification apparatus for preparing nucleic acid drugs according to claim 10, characterized in that, The first port may be selectively connected to the third port, and the second port may be selectively connected to the fourth port.

12. The purification apparatus for preparing nucleic acid drugs according to claim 10, characterized in that, The first port may be selectively connected to the fourth port.

13. The purification apparatus for preparing nucleic acid drugs according to claim 10, characterized in that, A bubble sensor is installed in the chromatography inlet pipeline; The equipment for preparing nucleic acid drugs also includes a bubble removal pipeline, which is connected to the chromatography inlet pipeline via a bubble trap.

14. The purification apparatus for preparing nucleic acid drugs according to claim 10, characterized in that, With two or more chromatography inlet lines provided, the two or more chromatography inlet lines are connected together to a static mixer and then to the chromatography main line, the static mixer having a bend in the flow channel.

15. The purification apparatus for preparing nucleic acid drugs according to claim 9, characterized in that, The pre-purification process module includes a magnetic bead purification process module, which includes... At least one magnetic bead is used for purification at the inlet end; The magnetic bead purification container has its inlet and outlet connected by pipelines to form a magnetic bead purification circulation loop; as well as A magnetic source, located at the magnetic bead purification container, is used to selectively adsorb magnetic beads.

16. The purification apparatus for preparing nucleic acid drugs according to claim 15, characterized in that, It also includes an oscillation mechanism connected to the magnetic bead purification container and driving the solution and magnetic beads inside the magnetic bead purification container to oscillate.

17. The purification apparatus for preparing nucleic acid drugs according to claim 15, characterized in that, The magnetic source includes a magnetic body and a magnetic source driving mechanism. The magnetic source driving mechanism is connected to the magnetic body to drive the magnetic body to move between adsorption and desorption positions.

18. The purification apparatus for preparing nucleic acid drugs according to claim 1, characterized in that, It also includes at least one mounting bracket, the mounting bracket comprising a frame, and a plurality of first connectors and a plurality of second connectors disposed on the frame, wherein the first connectors extend along a first direction, the second connectors extend along a second direction, and the first and second directions are alternately arranged.

19. The purification apparatus for preparing nucleic acid drugs according to claim 18, characterized in that, The first direction is vertical, the second direction is horizontal, and the bottom end of the first connector is also provided with a hook structure.

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