System and method for the production of mRNA

Abstract

A system and method for the production of mRNA pertain to the technical field of mRNA manufacturing, and they can shorten the manufacturing cycle while avoiding the degradation of an mRNA product due to the introduction of RNase by manual pipetting. The mRNA manufacturing system comprises: a PCR amplification device configured for DNA amplification; several test tubes, each configured to hold a single reagent; and at least one reaction device configured to enable the reaction of reagents; and specifically, the reaction device comprises: a test tube rack configured to hold reaction tubes; a semiconductor cooling plate connected to the test tube rack; a finned condenser positioned below the test tube rack; and a cooling fan positioned below the finned condenser.wherein the reagent tube and the reaction tube communicate with each other via a solenoid valve assembly and a peristaltic pump assembly, and the solenoid valve assembly and the peristaltic pump assembly are connected to a control device, and it is controlled by a control program of a higher-level computer.

Description

Cross-Reference to Related ApplicationsThe present disclosure claims priority to the Chinese patent application filed on Jul. 17, 2019, in the Chinese patent office having application no. 201910648409.7, and entitled "System and Method for Producing mRNA".Technical FieldThe present disclosure relates to the technical field of mRNA production, and more particularly relates to a system and method for producing mRNA.Background ArtmRNA vaccines are nucleic acid vaccines with immunity, safety, and flexibility. Compared to conventional vaccines, mRNA vaccines are distinguished by high safety, rapid research and development, low research and development costs, simple mass production, and ready-to-use application.In the production of mRNA vaccines, reaction materials in test tubes and reaction reagents in reaction tubes mostly contain enzymes to be stored in an environment of the temperature 4°C; moreover, the existing method for the production of mRNA vaccines requires several facilities and more labor, resulting in the relatively longer production cycle. Further, in conventional methods for producing mRNA, liquid replenishing and pipetting is manually realized by a pipette, whereby RNase is easily introduced, resulting in degradation of mRNA and causing low yield.Therefore, there are the following technical problems that the skilled person must solve strongly, such as providing a system and a method for producing mRNA to shorten the production cycle and at the same time avoid contamination due to manual pipetting.Subject Matter of the DisclosureIt is an object of the present disclosure to provide, for example, a system and a method for producing mRNA which can shorten the production cycle and at the same time avoid degradation of mRNA due to the introduction of RNase by manual pipetting.Embodiments of this disclosure are to be carried out as follows:A system for producing mRNA comprises: a PCR amplification device configured to amplify DNA; a plurality of test tubes, each test tube of which is configured to provide a single reagent; and at least one reaction device configured to allow reaction of reagents; and the reaction device comprises: a test tube rack configured to support test tubes; a semiconductor cooling plate in communication with the test tube rack; a finned cooler disposed below the test tube rack; and a cooling fan disposed below the finned cooler; wherein the test tubes and the reaction tube are in communication with each other via a solenoid valve assembly and a tube pump assembly, and the solenoid valve assembly and the tube pump assembly are in communication with a control device under control of a master computer.Optionally, the PCR amplification device comprises a PCR heating column that is simultaneously in communication with the solenoid valve assembly and the tube pump assembly and configured for amplification of template DNA.Optionally, the test tubes comprise a first test tube, a second test tube, a third test tube, and a fourth test tube; wherein the first test tube is configured to receive a capture reagent; the second test tube is configured to receive a transcription reagent; the third test tube is configured to receive an elution reagent; and the fourth test tube is configured to receive a washing reagent.Optionally, any form of wound fins, series fins, solder tails, and rolled fins are used for the fin cooler.Optionally, the reaction device further comprises a housing sheathed around the outside of the test tube rack and the semiconductor cooling plate and abutting the fin cooler; and an end of the reaction tube protrudes from the housing.Optionally, the housing and the fin cooler together define a cavity configured to receive the test tube rack.Optionally, the reaction device is equipped with a plunger pump assembly in communication with the reaction tube.Optionally, any of a single plunger pump, horizontal plunger pump, axial plunger pump, and radial plunger pump is used for the plunger pump assembly.Optionally, a plurality of reaction devices are present, the mRNA production system further comprises a magnetic attraction assembly, and each of two adjacent reaction devices among the plurality of reaction devices are communicated with each other through a common magnetic attraction assembly.Optionally, there are four reaction devices.Optionally, the magnetic attraction assembly includes an electric machine and a magnet connected to the electric machine, the electric machine configured to drive the magnet to reciprocate between adjacent reaction devices.Optionally, the system for producing mRNA in the present disclosure further comprises a waste liquid collection bottle in communication with the reaction tube and configured to collect the waste liquid in the reaction tube.Optionally, the waste liquid collection bottle is in communication with the reaction tube through the tube pump assembly and the solenoid valve assembly sequentially.Optionally, the system for producing mRNA further comprises a base, and the reaction device and the PCR amplification device are both in communication with the base.Optionally, the mRNA production system further comprises a support frame, wherein the support frame communicates with the base, a receiving space is provided between the support frame and the base, and the PCR amplification device is located in the receiving space; and the test tubes, the solenoid valve assembly, and the tube pump assembly are all in communication with the support frame.Optionally, the support frame comprises a head plate, a first side plate, and a second side plate, wherein the first side plate and the second side plate are both in communication with the head plate, and the first side plate and the second side plate are both in communication with the base; the test tubes, the solenoid valve assembly, and the tube pump assembly are all in communication with the head plate; and the controller is in communication with the second side plate.The embodiments of the present disclosure have the following advantageous effects, for example:The semiconductor cooling plate in the reaction apparatus is configured to cool or heat the test tube rack, the reaction tube is inserted into the test tube rack, and control over the temperature of the reaction tube is realized by the cooling fan and the fin cooler in combination with the semiconductor cooling plate, the storage temperature of the reagent thereof is 4°C, and the temperature required for the reaction of the reagent is quickly achievable. In addition, the test tubes and the reaction device are connected to each other inside the reaction tube through the solenoid valve assembly and the tube pump assembly, and the PCR amplification device is simultaneously communicated with the solenoid valve assembly and the tube pump assembly, whereby good control over the flow of a liquid can be realized by controlling ON or OFF of the solenoid valve assembly and the tube pump assembly. For this reason, degradation of mRNA due to introduction of RNase by manual pipetting can be effectively prevented, and the yield of mRNA is effectively increased; besides, DNA from the liquid is amplified during the flow by the PCR amplification device, and good amplification effect is obtained. Moreover, multiple components are integrated into the mRNA production system, potentially reducing the volume of the instrument, facilitating observation of the entire experiment process by experimenters and enabling a short production cycle.Moreover, the following advantages are obtained by using the semiconductor cooling plate: applicability in a limited space, high reliability and rapid refrigeration, no contamination due to refrigerant, and eliminating an additional cold source. By utilizing the Peltier effect of a semiconductor material, heat absorption and heat emission can be respectively realized quickly at both ends of a galvanic pair consisting of two different semiconductor materials connected in series when a direct current flows through the galvanic pair, thereby realizing the purpose of quick refrigeration and effectively shortening the manufacturing cycle.It is a further object of the present disclosure to provide a method for the production of mRNA which enables facilitated and rapid production of mRNA vaccines.An embodiment of this disclosure is to be implemented as follows:In the method for producing mRNA, the system for producing mRNA is used for production and production as described above under any item; and the method comprises the steps of: performing amplification of template DNA using the PCR amplification device; performing a biotin affinity assay using the solenoid valve assembly and the tube pump assembly; performing cap modification using the solenoid valve assembly and the tube pump assembly; performing tailing transcription using the solenoid valve assembly and the tube pump assembly; and performing purification of an mRNA product using the solenoid valve assembly and the tube pump assembly.The embodiment of the present disclosure has the following advantageous effects, for example:By controlling ON or OFF of the solenoid valve assembly and the tube pump assembly, good control over the flow of a liquid can be realized; for this reason, degradation of mRNA due to the introduction of RNase by manual pipetting can be effectively prevented, and the yield of mRNA is effectively increased, thereby facilitating and rapidly realizing the production of mRNA vaccines.Here, it is to be additionally explained that mRNA, also referred to as messenger RNA (messenger RNA), is obtained by transcription of a DNA strand as a template, usually comprises structures such as 5' untranslated region (UTR), open reading frame (ORF), and 3' UTR, etc., and is a kind of single-stranded ribonucleic acids which carry genetic information and can lead protein synthesis.Furthermore, DNA is also referred to as deoxyribonucleic acid, which is an organic compound having a complex molecular structure. As a component of the chromosomes, it is present in the nucleus. The function of this is to store the erb information. DNA molecules are huge and consist of nucleotides. The nitrogenous bases of nucleotides are adenine, guanine, cytosine, and thymine; and pentose is deoxyribose. In 1953 the structure of DNA from James Dewey Watson of the USA, Crick and Wilkins of the United Kingdom has been described: the structure of DNA is formed by a pair of polynucleotide chains wound around a common central axis. The sugar-phosphate chain is located on the outside of the helical structure with the bases directed inwardly. The two polynucleotide chains are joined by hydrogen bonding between the bases and form a quite stable combination.DESCRIPTION OF THE DRAWINGSIn order to more clearly describe technical solutions of the embodiments of this disclosure, the drawings necessary for the embodiments are briefly recited below; and it is to be understood that the following drawings show only some embodiments of this disclosure, and therefore should not be considered limitations on the scope, and other related drawings may be available to those skilled in the art from these drawings without expending inventive activity. FIG. 1 is a schematic structural diagram of a system for producing mRNA according to the present disclosure; FIG. 2 is an exploded schematic structural diagram of a reaction apparatus of the mRNA production system according to the present disclosure; FIG. 3 is a schematic flow diagram of a method for preparing mRNA according to the present disclosure; and Figure 4 is a schematic illustration of a system for automated and rapid synthesis of mRNA vaccines in accordance with the present disclosure.In the figures:1-reaction device; 11-reaction tube; 12-tube rack; 13-semiconductor cooling plate; 14-fin cooler; 15-cooling fan; 16-housing; 2-plunger pump assembly; 3-solenoid attracting assembly; 5-tube; 6-solenoid valve assembly; 7-tube pump assembly; 8-control device; 9-liquid waste collection bottle; 10-PCR heating column; 100-base; 200-support frame; 210-head plate; 220-first side plate; 230-second side plate; 300-base; 400-receiving space; and 500-PCR amplification device.Detailed Description of EmbodimentsIn order to clarify the objects, the technical solutions, and the advantages of the embodiments of the present disclosure, technical solutions in the embodiments of this disclosure will be described clearly and comprehensively below with reference to the drawings in the embodiments of this disclosure, and obviously the described embodiments are partial embodiments instead of all of the embodiments of the disclosure. In general, the assemblies of the embodiments of this disclosure described and illustrated herein in the drawings may be arranged and configured in a number of different configurations.Therefore, the following detailed description of the embodiments of this disclosure provided in the drawings should only represent selected embodiments of this disclosure, rather than limiting the claimed scope of the present disclosure. All other embodiments that would be available to those skilled in the art based on the embodiments in this disclosure without inventive activity fall within the scope of the disclosure.Note that like reference numerals and letters in the following drawings refer to like items, and thus an item is not to be further defined and explained in subsequent drawings as long as the item has been once defined in a drawing.In the explanation of this disclosure, it is to be explained that orientation or positional relationships indicated by terms such as "central", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", and "outer" based on the drawings are orientation or positional relationships or orientation or positional relationships in which a product of this disclosure is conventionally placed, and the terms are only for the convenience of explanation of the disclosure and for the convenience of explanation, instead of suggesting or suggesting that the mentioned device has a specific orientation and should be constructed and operated in a specific orientation, and therefore should not be understood as limitations of the disclosure. Further, terms such as "first", "second", and "third" are only for the purpose of different explanation, and should not be interpreted as indicating importance in relativeness.Further, terms such as "horizontal", "vertical", and "overhanging", etc. do not require a component to be arranged absolutely horizontally or overhanging, but may be slightly inclined. For example, the term "horizontal" only refers to a direction that is more horizontal than "vertical" rather than to an absolutely horizontal structure, but this structure may be slightly inclined.In explaining this disclosure, it is to be further explained that terms such as "arranging", "mounting", "connecting", and "connecting" should be understood in a broad sense unless otherwise expressly stated and defined. For example, it could be either a fixed connection, or a removable connection, or an integrated connection; it could be either a mechanical connection, or an electrical connection; and it could be either a direct connection, or an indirect connection through an intermediate member, or an internal communication between two elements. Those skilled in the art could understand the specific meanings of the above terms in this disclosure according to specific circumstances.Some embodiments of the present disclosure will be described in detail below with reference to the accompanying drawings. The following embodiments and the features in the embodiments can be combined with each other without contradiction.Referring to FIGS. 1 and 2, the present disclosure provides a system for producing mRNA, comprising: a PCR amplification device 500, at least one reagent tube 5, and at least one reaction device 1, the PCR amplification device 500 is configured to amplify DNA; the number of the reagent tubes 5 is adjusted as needed, for example, there are a plurality of the reagent tubes 5, and each of the reagent tubes 5 is configured to provide a single reagent; and the reaction device 1 is configured to enable reaction of reagents. Optionally, the reaction device 1 includes a reaction tube 11, a test tube rack 12, a semiconductor cooling plate 13, a fin cooler 14, and a cooling fan 15. the test tube rack is configured to support reaction tubes 11, the test tube rack 12 communicates with the semiconductor cooling plate 13, the fin cooler 14 is disposed below the test tube rack 12, and the cooling fan 15 is disposed below the fin cooler 14; here, the test tubes 5 and the reaction tube 11 communicate with each other via a solenoid valve assembly 6 and a tube pump assembly 7, and the solenoid valve assembly 6 and the tube pump assembly 7 both communicate with a control device 8, and it is controlled by a control program of a host computer.Optionally, the mRNA production system further comprises a waste liquid collection bottle 9 in sequential communication with the reaction tube 11 through the tube pump assembly 7 and the solenoid valve assembly 6 and configured to collect the waste liquid in the reaction tube 11. By disposing the liquid waste collection bottle 9, not only the collection of the liquid waste generated from the reaction tube 11 is facilitated, but the environmental pollution due to the liquid waste can be effectively reduced.Optionally, the mRNA manufacturing system further comprises a base 100 and a support frame 200 disposed on the base 100, wherein the base 100 could be a flat plate structure, the support frame 200 is disposed on a plate surface of the base 100, and the support frame 200 and the base 100 can be firmly connected to each other by using a mode such as a screw, a bolt, a rivet, or by welding, wherein a receiving space 400 is formed between the support frame 200 and the base 100. The reaction device 1 is disposed on the base 100. Optionally, the cooling fan 15 of the reaction device 1 is connected to the base 100, and the reaction device 15 may be connected to the base 100 by a screw, a bolt, or a rivet.Optionally, the support frame 200 includes a head plate 210, a first side plate 220, and a second side plate 230, wherein the first side plate 220 and the second side plate 230 are both connected to the head plate 210, the first side plate 220 and the second side plate 230 are simultaneously connected to a bottom plate, the head plate 210 is parallel to the bottom plate, and the head plate 210 is spaced apart from the bottom plate. Optionally, the first side plate 220 and the second side plate 230 are vertical. On the head plate 2105, a socket 300 configured to plug the tube 5 is disposed; and the number of the sockets 300 corresponds to the number of the tubes 5, and the two are respectively in one-to-one plug cooperation. The solenoid valve assembly 6, the tube pump assembly 7, and the liquid waste collection bottle 9 are all disposed on the top plate 210, and the control device 8 is disposed on the second side plate 230.Referring to FIG. 1, in the present disclosure, the PCR amplification device 500 may include a PCR heating column 10 that simultaneously communicates with the solenoid valve assembly 6 and the tube pump assembly 7, thereby enabling amplification of the DNA by the PCR amplification device 500. In other words, heat exchange takes place between the heated PCR heating column 10 and a reagent flowing in the solenoid valve assembly 6 and the tube pump assembly 7, so that the reagent is heated, and then amplification of DNA is realized.Here, in addition, it is to be explained that polymerase chain reaction is abbreviated as PCR (polymerase chain reaction), and PCR is a method for in vitroenzymatic synthesis of specific DNA fragments, reactions such as high temperature denaturation, low temperature annealing (renaturation), and extension at an adjusted temperature, etc. are cycled and periodically performed so that target DNA is rapidly amplified; and therefore, PCR has peculiarities such as high specificity, high sensitivity, easy operation, and time saving.In the present disclosure, the tubes 5 may include a first tube, a second tube, a third tube, and a fourth tube; optionally, the first tube may be configured to receive a capture reagent, the second tube may be configured to receive a transcription reagent, the third tube may be configured to receive an elution reagent, and the fourth tube may be configured to receive a washing reagent.It is to be explained that the number of tubes 5 need not be limited to 4 as described above, for example, 3 or 5 tubes 5 etc. could be present.In the present disclosure, any form of wound fins, series fins, solder tails, and rolled fins may be used for the fin cooler 14.Referring to FIGS. 1 and 2, the reaction device 1 in the present disclosure may further include a housing 16 that may be sheathed around the outside of the test tube rack 12 and the semiconductor cooling plate 13 and abut on the fin cooler 14. The housing 16 and the fin cooler 14 collectively define a cavity in which the temperature is comparatively stable and in which the test tube rack 12 is located; in addition, the reaction tube 11 is inserted into the test tube rack 12 and one end thereof protrudes from the housing 6, thereby promoting maintenance of the temperature of the reaction tube 11 and facilitating the production of mRNA.Referring to FIGS. 1 and 2, the reaction device 1 in the present disclosure may be equipped with a plunger pump assembly 2 disposed on the base 100 and in communication with the reaction tube 11, thereby replacing conventional agitation and mixing with the plunger pump assembly 2, then effectively reducing the volume of the instrument and reducing the complexity of the instrument, and reducing the cost.In the present disclosure, any of a single plunger pump, a horizontal plunger pump, an axial plunger pump, and a radial plunger pump may be used for the plunger pump assembly 2.Referring to FIGS. 1 and 2, in the present disclosure, the number of the reaction device 1 may be set to four, two reaction devices may be configured to realize a magnetic attraction function, and the two reaction devices 1 to be realized the magnetic attraction function may be connected to each other by a common magnetic attraction assembly 3.Referring to FIGS. 1 and 2, the magnetic attraction assembly 3 in the present disclosure may include an electric machine and a magnet connected to this electric machine, the electric machine being disposed on the base 100, and the electric machine may drive the magnet to move in parallel by rotating, that is, the magnet may reciprocate between the two reaction devices 1 to realize the magnetic attraction function under the drive of the electric machine. Both reaction devices 1 share the same magnet, effectively reducing the volume of the instrument and reducing the cost.The system for producing mRNA provided in the present disclosure has the following advantageous effects, for example:The semiconductor cooling plate 13 in the reaction apparatus 1 is configured to cool or heat the test tube rack 12, the reaction tube 11 is inserted into the test tube rack 12, and the temperature of the reaction tube 11 is controlled by the cooling fan 15 and the fin cooler 14 in combination with the semiconductor cooling plate 13, thereby ensuring a temperature of the reaction tube 11 stabilized at the required temperature. In addition, the PCR amplification device 500, the test tubes 5, and the reaction device 1 are connected to each other through the solenoid valve assembly 6 and the tube pump assembly 7, and the PCR amplification device 500 simultaneously communicates with the solenoid valve assembly 6 and the tube pump assembly 7, whereby good control over the flow of a liquid can be realized by controlling ON or OFF of the solenoid valve assembly 6 and the tube pump assembly 7. For this reason, degradation of mRNA due to introduction of RNase by manual pipetting can be effectively prevented, and the yield of mRNA is effectively increased; besides, DNA from the liquid is amplified during the flow by the PCR amplification device 500, and good amplification effect is obtained. Moreover, multiple components are integrated into the mRNA production system, potentially reducing the volume of the instrument, facilitating observation of the entire experiment process by experimenters and enabling a short production cycle.Moreover, the following advantages are obtained by using the semiconductor cooling plate 13: applicability in a limited space, high reliability and rapid refrigeration, no contamination due to refrigerant, and eliminating an additional cold source. By utilizing the Peltier effect of a semiconductor material, heat absorption and heat emission can be respectively realized quickly at both ends of a galvanic pair consisting of two different semiconductor materials connected in series when a direct current flows through the galvanic pair, thereby realizing the purpose of quick refrigeration and effectively shortening the manufacturing cycle.Referring to FIG. 3, the present disclosure provides a method for producing mRNA using the system for producing mRNA mentioned in the present disclosure for production and production of mRNA; and the method comprises the following steps:Step S 1: Performing amplification of template DNA using the PCR amplification device; Step S 2: Performing a biotin affinity assay in the reaction device 1 using the solenoid valve assembly 6 and the tube pump assembly 7; Step S 3: Performing cap transcription in the reaction device 1 using the solenoid valve assembly 6 and the tube pump assembly 7; Step S 4: Performing a tailing modification in the reaction device 1 using the solenoid valve assembly 6 and the tube pump assembly 7; and Step S 5: Performing purification of an mRNA product in the reaction device 1 using the solenoid valve assembly 6 and the tube pump assembly 7.During practical operations, the following specific steps may be included, as shown in FIG. 4:Step S1: Amplification for 0.5 ml using a PCR amplification device:maintaining temperature 95°C for 5 minutes - [maintaining temperature 95°C for 40 s - maintaining temperature 58°C for 45 s - maintaining temperature 72°C for 1 minute] (30 cycles) - maintaining temperature 72°C for 10 minutes;Step S21: 0.5 ml amplification reagent - opening only valve A and pump A - 2.5 ml capture reagent:Shake and maintain [magnetic adsorption] 3 ml of supernatant for 45 minutes to 1 hour at 30°C - open only valve F and pump F - waste liquid collecting bottle [magnetic removal];Step S22: 2.5 ml capture reagent - opening valve B and pump A only - capture reagent:Shake [Magnetic adsorption] 2.5 ml of supernatant - Open only valve F and pump F - Waste liquid collection bottle [Magnetic removal] (repeating for 2 times);Step S23: 2.5 ml eluent - opening valve M only, valve D, and pump A - capture reagent:Shake [Magnetic adsorption] 2.5 ml of supernatant - Open only valve F and pump F - Liquid waste collection bottle [Magnetic removal];Step S31: 0.5 ml of transcription reagent - opening of valve C and pump A only - capture reagent:Shake and maintain for 4 hours [magnetic adsorption] at 37°C;Step S32: 0.5 ml capture reagent (after transcription) and supernatant - opening valve E and pump B only - 30 μl DNase;Shake and maintain for 15 minutes [magnetic adsorption] at 37°C;Step S4: Add 0.53 ml DNase and supernatant - open valve G and pump C only - 0.54 ml modifying reagent;maintaining for 1 hour at 37°C, and rapidly heating to 65°C and maintaining for 5 minutes;Step S51: Addition of 1.07 ml of modifying reagent and supernatant - Opening of valve H and pump D only - 1 ml of cleaning reagent:Shake and maintain for 5-10 minutes [magnetic adsorption] at room temperature;Step S52: Addition of 2.07 ml of cleaning reagent and supernatant - Opening of only valve L and pump G - Liquid waste collection bottle [magnetic removal];Step S53: 2 ml of washing liquid - opening only valve J and pump D - cleaning reagent:Shake [Magnetic adsorption] 2 ml of supernatant - opening only valve L and pump G - Liquid waste collection bottle [Magnetic removal] (repeating for 2 times);Step S54: 0.2 ml eluent - opening valve N only, valve I, and pump D - cleaning reagent:Maintain for 2 minutes and shake [magnetic adsorption] 0.2 ml of supernatant at 73°C - open valve K and pump E only - product [magnetic removal] (repeat for 3 times).mRNA vaccines - 10 dosesTest tubes: Reaction tubesCapture reagent: 5 ml 0.72 M binding buffer at normal temperature Amplification reagent: 0.5 ml PCRTranscription reagent: 0.5 ml cap transcription system at 4°C capture reagent: 2.5 ml resuspended streptavidin magnetic beads after 3 times washing 1 ml 0.72 M binding bufferEluent: 3.5 ml of 0.1% DEPC water at normal temperature DNase: 30 μlWash liquor: 4 ml washing at 4°C Modifying reagent: 0.54 ml Tailing transcription systemCleaning reagent: 1 ml of oligo d(T) magnetic beads resuspended from 100 μl after a single washing by 100 μl binding.The present disclosure has the following advantageous effects, for example: by controlling ON or OFF of the solenoid valve assembly 6 and the tube pump assembly 7, good control over the flow of a liquid can be realized, for this reason, degradation of mRNA due to the introduction of RNase by manual pipetting can be effectively prevented, and the yield of mRNA is effectively increased, thereby facilitating and rapidly realizing the production of mRNA vaccines.Here, it is to be additionally explained that the enzyme DNase, namely deoxyribonuclease, is an endonuclease capable of digesting single-stranded or double-stranded DNA to produce monodeoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides. RNase is an enzyme which acts only on RNA and is widely used in the environment, has very good stability, and has no sequence specificity, resulting in unprotected RNA being easily degraded therefrom.Although only preferred embodiments of the present disclosure are provided and are not configured to limit the disclosure, the present disclosure may be variously changed and modified by those skilled in the art. Any changes, equivalent substitutions, and alterations made within the spirit and principle of the disclosure should fall within the scope of the disclosure.Industrial applicability:In summary, the present disclosure provides a system and method for producing mRNA that enables a high yield of mRNA.

Claims

A system for producing mRNA, characterized in that the system comprises: a PCR amplification device configured to amplify DNA; a plurality of test tubes, each test tube being configured to provide a single reagent; and at least one reaction device configured to allow reaction of reagents; the reaction device comprises: a test tube rack configured to support test tubes; a semiconductor cooling plate in communication with the test tube rack; a fin cooler disposed below the test tube rack; and a cooling fan disposed below the fin cooler; the test tubes and the reaction tube are in communication with each other via a solenoid valve assembly and a tube pump assembly, and the solenoid valve assembly and the tube pump assembly are in communication with a control device under control of a master computer.The system for producing mRNA of claim 1, characterized in that the PCR amplification device comprises a PCR heating column that is simultaneously in communication with the solenoid valve assembly and the tube pump assembly and configured for amplification of template DNA.The system for producing mRNA according to claim 1 or 2, characterized in that the test tubes comprise a first test tube, a second test tube, a third test tube, and a fourth test tube; the first test tube is configured to accommodate a capture reagent; the second test tube is configured to accommodate a transcription reagent; the third test tube is configured to accommodate an elution reagent; and the fourth test tube is configured to accommodate a washing reagent.The system for producing mRNA according to any one of claims 1-3, characterized in that any form of wound fins, series fins, solder tails, and rolled fins is used for the fin cooler.The mRNA production system according to any one of claims 1-4, characterized in that the reaction device further comprises a housing sheathed around the outside of the test tube rack and the semiconductor cooling plate and communicating with the fin cooler; and an end of the reaction tube protrudes from the housing.The mRNA production system of any one of claims 1-4, characterized in that the housing and the air fin cooler together define a cavity configured to receive the test tube rack.The mRNA production system of any one of claims 1-6, characterized in that the reaction device is equipped with a plunger pump assembly in communication with the reaction tube.The mRNA production system of claim 7, characterized in that any one of single plunger pump, horizontal plunger pump, axial plunger pump, and radial plunger pump is used for the plunger pump assembly.The mRNA production system according to any one of claims 1-8, characterized in that a plurality of reaction devices are present, the mRNA production system further comprises a magnetic attraction assembly, and every two adjacent reaction devices among the plurality of reaction devices are communicated with each other through a common magnetic attraction assembly.The system for producing mRNA according to claim 9, characterized in that four reaction devices are present.The mRNA manufacturing system of claim 9 or 10, characterized in that the magnetic attraction assembly comprises an electric machine and a magnet connected to the electric machine, wherein the electric machine is configured to drive the magnet to reciprocate between adjacent reaction devices.The system for producing mRNA according to any one of claims 1-11, characterized in that a waste liquid collection bottle is still included, which is in communication with the reaction tube and is configured to collect the waste liquid in the reaction tube.The mRNA production system of claim 12, wherein the waste liquid collection bottle is in communication with the reaction tube through the peristaltic pump assembly and the solenoid valve assembly in sequence.The mRNA production system of any one of claims 1-13, characterized in that the mRNA production system further comprises a base, and the reaction device and the PCR amplification device are both in communication with the base.The mRNA production system of claim 14, characterized in that the mRNA production system further comprises a support frame, the support frame communicating with the base, a receiving space is provided between the support frame and the base, and the PCR amplification device is located in the receiving space; and the test tubes, the solenoid valve assembly, and the tube pump assembly are all communicating with the support frame.The mRNA manufacturing system of claim 15, characterized in that the support frame comprises a head plate, a first side plate, and a second side plate, wherein the first side plate and the second side plate are both in communication with the head plate, and the first side plate and the second side plate are both in communication with the base; the test tubes, the solenoid valve assembly, and the tube pump assembly are all in communication with the head plate; and the controller is in communication with the second side plate.A method for producing mRNA, characterized in that a system for producing mRNA according to any one of claims 1-16 as described above is used for production and production; and the method specifically comprises the steps of: performing amplification of template DNA using the PCR amplification device; performing a biotin affinity assay using the solenoid valve assembly and the tube pump assembly; performing cap transcription using the solenoid valve assembly and the tube pump assembly; performing tailing modification using the solenoid valve assembly and the tube pump assembly; and performing purification of an mRNA product using the solenoid valve assembly and the tube pump assembly.

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