In vitro transcription process module

Abstract

The application relates to the pharmaceutical technical field, and provides an in vitro transcription process module, which comprises a first pipetting piece and a first pipetting driving mechanism in cooperation connection. The first pipetting piece is driven to move by the first pipetting driving mechanism, so as to pipette at least two reactants into a reaction vessel. Then, the reaction vessel is driven to reciprocate by a mixing unit, and heat is transmitted to the reaction vessel by a temperature control mechanism of the mixing unit, so as to realize enzyme reaction. Therefore, efficient in vitro transcription operation is realized. In addition, at least one premixing can be carried out before the enzyme reaction, so that the effect and efficiency of in vitro transcription are improved.

Description

Technical Field

[0001] This application relates to the field of pharmaceutical technology, specifically to an in vitro transcription process module. 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 drugs prepared with mRNA as the core component. They regulate protein expression by transmitting genetic information and are currently mainly used in vaccine development (such as COVID-19 vaccines), gene therapy, and protein replacement therapy.

[0003] In the production process of mRNA stock solution, the existing in vitro transcription method involves adding various reagents to a reaction vessel sequentially and stirring them together. This not only results in low mixing efficiency but also makes the solution susceptible to contamination during the transfer process. Summary of the Invention

[0004] This application provides an in vitro transcription process module to improve the efficiency of in vitro transcription.

[0005] This application provides an in vitro transcription process module, including a reactor dish; a transfer unit including a first transfer element and a first transfer driving mechanism connected in cooperation, the first transfer driving mechanism being used to drive the first transfer element to move, so as to transfer at least two reactants into the reactor dish; and a mixing unit including a carrier, a mixing driving mechanism and a temperature control mechanism, the reactor dish being detachably fixed to the carrier, the mixing driving mechanism being connected to the carrier to drive the carrier to reciprocate, and the temperature control mechanism being connected to the reactor dish to transfer heat to the reactor dish.

[0006] In some embodiments, the in vitro transcription process module further includes a premixing unit disposed upstream of the mixing unit. The premixing unit includes a premixing container and a premixing assembly, including 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.

[0007] In some embodiments, the reactant is loaded into a reactant container before being removed by the first transfer member, and the reactant container of any of the reactants is used as the premix container.

[0008] In some embodiments, the carrier reciprocates along a first trajectory, and the premixed carrier reciprocates along a second trajectory, wherein the first trajectory and the second trajectory are different.

[0009] In some embodiments, the reactant is loaded into a reactant container before being removed by the first transfer member, the reactant container including a lid; The transfer unit further includes a second transfer member and a second transfer driving mechanism that are connected in conjunction. The second transfer driving mechanism is used to drive the second transfer member to move in order to transfer the reactant container and / or the cover.

[0010] In some embodiments, the transfer unit further includes a clamping mechanism, which includes a clamping member and a clamping drive mechanism that are connected in a mating manner. When the second transfer member transfers the reactant container to the clamping member, the clamping drive mechanism is used to drive the clamping member to move so as to clamp and fix the reactant container.

[0011] In some embodiments, the first pipette includes a pipette tip and a pressure source connected to the pipette tip for creating a negative pressure to adsorb the reactants. The pressure source is also used to intermittently create a negative pressure to mix the reactants.

[0012] In some embodiments, the hybrid drive mechanism includes a power source and an eccentric transmission structure that are connected in a mating manner. The eccentric transmission mechanism includes a drive eccentric wheel that is driveably connected to the output shaft of the power source. An eccentrically configured first eccentric transmission member is connected to the drive eccentric wheel, and the first eccentric transmission member is used to connect the carrier.

[0013] In some embodiments, the in vitro transcription process module further includes a passive eccentric wheel, which is rotatably mounted on a base. An eccentrically mounted second eccentric transmission member is connected to the passive eccentric wheel, and the second eccentric transmission member is also used to connect to the carrier.

[0014] In some embodiments, the mixing unit further includes a tilting component connected to one side of the carrier to drive one side of the carrier to lift, thereby facilitating the removal of the completed reaction solution.

[0015] In some embodiments, the in vitro transcription process module further includes a buffer container for fluid connection to the reactor vessel, the buffer container being detachably mounted on a base, the buffer container having a magnetic rotor disposed therein, and the base having a magnetic drive unit for generating magnetic force to drive the magnetic rotor to rotate and mix the drug solution in the buffer container.

[0016] This application offers the following advantages: It provides an in vitro transcription process module, comprising a first transfer element and a first transfer driving mechanism connected in cooperation. The first transfer driving mechanism drives the first transfer element to move, transferring at least two reactants into a reactor dish. Then, a mixing unit drives the reactor dish to reciprocate, and a temperature control mechanism of the mixing unit transfers heat to the reactor dish to achieve the enzyme reaction. This achieves highly efficient in vitro transcription. Furthermore, at least one pre-mixing can be performed before the enzyme reaction, thereby improving the effectiveness and efficiency of in vitro transcription. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 An exemplary schematic diagram of an in vitro transcription process module is shown.

[0019] Figure 2 An exemplary top view of an in vitro transcription process module is shown.

[0020] Figure 3 An exemplary schematic diagram of a transfer unit is shown.

[0021] Figure 4 Example shown Figure 2 A magnified view of part A in the diagram.

[0022] Figure 5 An exemplary schematic diagram of a hybrid unit is shown.

[0023] Figure 6 An exemplary schematic diagram of a hybrid drive mechanism is shown.

[0024] Figure 7 An exemplary schematic diagram of the structure of an insulated container is shown.

[0025] Figure 8 An example is shown. Figure 7 Top half-section view.

[0026] Figure 9 Example shown Figure 8 A schematic diagram of the PP cross-section.

[0027] Figure 10 An exemplary schematic diagram of a buffer container and a base in conjunction is shown.

[0028] Figure 11 An exemplary schematic diagram of a buffer container is shown.

[0029] Figure 12 An exemplary schematic diagram of a base for adapting a buffer container is shown.

[0030] Figure 13 An exemplary schematic diagram of the steps of an in vitro transcription method is shown.

[0031] Explanation of reference numerals in the figure: 11-Working platform, 12-Reaction dish, 100-Pipette unit, 110-First pipette drive mechanism, 111-First lateral movement mechanism, 112-First rotation mechanism, 113-First vertical movement mechanism, 120-First pipette component, 121-Pipette tip, 122-Pressure source, 123-Tip mounting part, 130-Second pipette drive mechanism, 131-Second lateral movement mechanism, 132-Second rotation mechanism, 133-Second vertical movement mechanism, 140-Second pipette component, 150-Clamping mechanism, 151-Clamping component, 160-Refrigeration chamber, 170-Waste liquid chamber, 180-Buffer container, 1801-Magnetic rotor, 1802-Second fixed... Positioning structure, 181-base, 1811-magnetic drive component, 1812-first positioning structure, 1813-weighing component, 191-refrigerated preparation storage area, 192-room temperature preparation storage area, 193-pipette tip storage area, 200-mixing unit, 210-bearing component, 211-heat insulation seat, 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 component, 241-tilting motor, 242-reducer, 243-connecting frame, 244-positioning component. Detailed Implementation

[0032] 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.

[0033] 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.

[0034] Please see Figure 1 and Figure 2 This application provides an in vitro transcription process module, which may include a transfer unit 100 and a mixing unit 200.

[0035] 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.

[0036] 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.

[0037] 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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] For example, the pipette tip 121 can be a disposable tip, and different pipette tips 121 can be used when transferring different reactants.

[0051] 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.

[0052] 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.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 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.

[0059] 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.

[0060] For example, the second transfer member 140 may be a structure such as an electric or pneumatic gripper or a robotic arm.

[0061] 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.

[0062] 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.

[0063] 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.

[0064] 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).

[0065] 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.

[0066] 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.

[0067] Please refer to the following for details. Figure 6The 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] Please refer to the following: Figure 5The 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.

[0072] 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.

[0073] 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.

[0074] 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.

[0075] 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.

[0076] 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.

[0077] 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.

[0078] 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.

[0079] 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.

[0080] In some embodiments, please refer to Figure 10 The 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.

[0081] 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 it 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 medicine liquid.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] 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.

[0090] See another aspect of this application. Figure 13 The embodiments of this application also provide an in vitro transcription method, which can employ the in vitro transcription process module provided in any of the foregoing embodiments.

[0091] Here, the in vitro transcription method includes the following steps S100 to S200.

[0092] In S100, at least two reactants are transferred into the reactor vessel 12 by the transfer unit 100.

[0093] 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.

[0094] 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.

[0095] 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.

[0096] 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.

[0097] 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.

[0098] This application provides an in vitro transcription process module, including 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 a reactor dish 12. Then, a mixing unit 200 drives the reactor dish 12 to reciprocate, and a 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.

[0099] In addition, premixing can be performed at least once before the enzyme reaction, thereby improving the effectiveness and efficiency of in vitro transcription.

[0100] 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. Example

[0101] Here, the example of using two reactants is used for illustration. It is understood that unless otherwise stated, the order of the steps described in this application does not represent a limitation on their order.

[0102] 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.

[0103] 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.

[0104] After mixing, the first transfer member 120 transfers the reactant to the reactant container of another reactant, and repeatedly draws in and releases at least two reactants in the reactant container by intermittently forming a negative pressure, thereby achieving mixing.

[0105] The second transfer member 140 transfers a reactant container containing two reactants to a premixing unit, which shakes the reactant container to achieve premixing.

[0106] 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.

[0107] After premixing, the first transfer member 120 transfers the drug solution from the reactant container to the reactor dish 12, and the second transfer member 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.

[0108] 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.

[0109] 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.

[0110] After the lid is removed, the tilting component 240 tilts the reactor vessel 12 to allow the solution to be removed. Example

[0111] When more reactants are required according to process requirements, more reactants can be added through the second transfer member 140 according to process requirements.

[0112] 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.

[0113] 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.

[0114] 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.

[0115] The in vitro transcription process module provided in this application has been described in detail above. 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 method 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. An in vitro transcription process module, characterized in that, include, Reactor dish; The transfer unit includes a first transfer member and a first transfer driving mechanism that are connected in a mating manner. The first transfer driving mechanism is used to drive the first transfer member to move so as to transfer at least two reactants into the reactor dish. as well as The mixing unit includes a support, a mixing drive mechanism, and a temperature control mechanism. The reactor dish is detachably fixed to the support. The mixing drive mechanism is connected to the support to drive the support to reciprocate. The temperature control mechanism is connected to the reactor dish to transfer heat to the reactor dish.

2. The in vitro transcription process module according to claim 1, characterized in that, It also includes a premixing unit disposed upstream of the mixing unit, the premixing unit comprising, Premixed containers; as well as A premixed assembly includes a premixed carrier and a premixed drive mechanism, the premixed drive mechanism being connected to the premixed carrier to drive the premixed carrier to reciprocate.

3. The in vitro transcription process module according to claim 2, characterized in that, Before being removed by the first transfer member, the reactants are loaded into reactant containers, and the reactant container of any of the reactants is used as the premix container.

4. The in vitro transcription process module according to claim 2, characterized in that, The carrier moves back and forth along a first trajectory, and the premixed carrier moves back and forth along a second trajectory, the first trajectory being different from the second trajectory.

5. The in vitro transcription process module according to claim 1 or 2, characterized in that, Before being removed by the first transfer member, the reactant is loaded into a reactant container, which includes a lid; The transfer unit further includes a second transfer member and a second transfer driving mechanism that are connected in conjunction. The second transfer driving mechanism is used to drive the second transfer member to move in order to transfer the reactant container and / or the cover.

6. The in vitro transcription process module according to claim 5, characterized in that, The transfer unit further includes a clamping mechanism, which includes a clamping member and a clamping drive mechanism that are connected in a mating manner. When the second transfer member transfers the reactant container to the clamping member, the clamping drive mechanism is used to drive the clamping member to move so as to clamp and fix the reactant container.

7. The in vitro transcription process module according to claim 1, characterized in that, The first transfer device includes a pipette tip and a pressure source. The pressure source is connected to the pipette tip to create a negative pressure for adsorbing the reactants. The pressure source is also used to intermittently create a negative pressure to agitate the reactants by changing the air pressure.

8. The in vitro transcription process module according to claim 1, characterized in that, The hybrid drive mechanism includes a power source and an eccentric transmission structure that are connected in a cooperating manner. The eccentric transmission structure includes a drive eccentric wheel that is driveably connected to the output shaft of the power source. An eccentrically configured first eccentric transmission member is connected to the drive eccentric wheel, and the first eccentric transmission member is used to connect the load-bearing member.

9. The in vitro transcription process module according to claim 8, characterized in that, It also includes a passive eccentric wheel, which is rotatably mounted on a base. A second eccentric transmission component is connected to the passive eccentric wheel, and the second eccentric transmission component is also used to connect the load-bearing component.

10. The in vitro transcription process module according to claim 1, characterized in that, The mixing unit also includes a tilting component connected to one side of the carrier to drive one side of the carrier to lift, thereby facilitating the removal of the solution that has completed the reaction.

11. The in vitro transcription process module according to claim 1, characterized in that, It also includes a buffer container for fluid connection to the reactor vessel, the buffer container being detachably mounted on a base, the buffer container having a magnetic rotor inside, and the base having a magnetic drive unit for generating magnetic force to drive the magnetic rotor to rotate and mix the drug solution in the buffer container.

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