MRNA (messenger ribonucleic acid) production device
By using a heat-conducting fluid-driven shaking plate to optimize reaction mixture mixing, the problem of mRNA structure destruction during IVT reaction amplification was solved, enabling large-scale production of high-quality mRNA.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-21
- Publication Date
- 2026-03-03
AI Technical Summary
In current IVT reaction scale-up processes, traditional mixing techniques result in significant internal shear forces in the solution, which damage the mRNA structure and limit the large-scale industrial production of mRNA.
An mRNA production device is adopted, which includes a production tank, heat transfer fluid, linkage components and solution disturbance components. The flow of heat transfer fluid drives the shaking of the placement plate, which indirectly affects the flexible reaction bag, so as to achieve uniform mixing of the reaction solution and reduce the damage to the mRNA structure.
While maintaining the homogeneity of the reaction system, the yield and quality of mRNA were improved, enabling the industrial-scale production of mRNA.
Smart Images
Figure CN223963479U_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the technical field of mRNA production and manufacturing, and in particular relates to an mRNA production device. Background Technology
[0002] Messenger ribonucleic acid (mRNA) is a single-stranded ribonucleic acid molecule that efficiently translates its genetic information into corresponding proteins on ribosomes within cells. mRNA vaccines and mRNA drugs deliver RNA molecules encoding specific antigen proteins through specific delivery systems, expressing the corresponding antigens in vivo or modulating gene expression within cells, thereby stimulating a specific immune response or altering metabolism to achieve disease prevention and / or treatment. mRNA vaccines and mRNA drugs respond rapidly to changes in diseases and pathogens and have high safety profiles, thus showing broad application prospects.
[0003] Currently, mRNA production mainly involves the design and production of linear plasmids, in vitro transcription (IVT) of mRNA, mRNA purification, and encapsulation. IVT is a crucial step in mRNA production, involving the transcription and synthesis of mRNA using linear plasmids as templates in a cell-free system. In scale-up IVT reactions, mixing is essential to ensure the homogeneity of the reaction system and achieve better preparation results. However, in scaled-up IVT reaction systems, traditional mixing techniques often lead to significant internal shear forces in the solution, severely damaging the mRNA structure and causing preparation failure. This limits the large-scale industrial production of mRNA and presents significant limitations. Utility Model Content
[0004] The purpose of this invention is to resolve the contradiction between the homogeneity of the reaction system and the integrity of the mRNA structure in existing IVT reaction scale-up processes, and to provide an mRNA production device. This mRNA production device can maintain good homogeneity of the IVT reaction system while reducing damage to the mRNA structure, thereby producing large quantities and high-quality mRNA, which is beneficial for the industrial-scale production of mRNA.
[0005] Specifically, the mRNA production apparatus provided by this utility model includes: a production tank, wherein a heat-conducting liquid is disposed inside the production tank; a linkage component, wherein the linkage component is disposed inside the production tank, the linkage component includes an elastic connector and a placement plate, the elastic connector is connected to the peripheral wall of the production tank, and the placement plate is connected to the elastic connector to be suspended inside the production tank; a flexible reaction bag, wherein the flexible reaction bag is disposed on the placement plate, and the flexible reaction bag and the placement plate are immersed in the heat-conducting liquid; and a solution agitation component, wherein the solution agitation component is used to drive the flow of the heat-conducting liquid to cause the placement plate to shake, and the solution agitation component maintains a non-contact state with the flexible reaction bag and the placement plate.
[0006] Furthermore, the production tank includes a tank body and a top cover; the top cover is connected to one end of the elastic connector, and the shelf is connected to the other end of the elastic connector, so that the elastic connector is in a stretched state.
[0007] Furthermore, the bottom wall of the production tank is connected to one end of the elastic member, and the placement plate is connected to the other end of the elastic connector, so that the elastic connector is in a compressed state.
[0008] Furthermore, the shelf is tilted.
[0009] Furthermore, the shelf is a grid board.
[0010] Furthermore, the solution disturbance component is a magnetic stirring component, which includes a magnetic stirring platform and a magnetic rotor. The production tank is disposed on the magnetic stirring platform, and the magnetic rotor is disposed inside the production tank to drive the flow of the heat transfer liquid. The magnetic rotor is kept in a non-contact state with the flexible reaction bag and the placement plate.
[0011] Furthermore, the flexible reaction bag is disposed on the side of the placement plate opposite to the magnetic rotor.
[0012] Furthermore, the solution agitation component is a mechanical stirring component, which includes a stirring rod and a stirring motor. The stirring motor drives the stirring rod to rotate. The stirring rod is inserted into the production tank to drive the flow of the heat transfer liquid, and the stirring rod remains in a non-contact state with the flexible reaction bag and the placement plate.
[0013] Furthermore, the flexible reaction bag is disposed on the side of the placement plate opposite to the stirring rod.
[0014] Furthermore, a temperature controller is installed inside the production tank.
[0015] Beneficial effects:
[0016] The mRNA production apparatus provided by this invention includes a production tank containing a heat-conducting liquid, a linkage component disposed within the production tank, and a solution agitation component for driving the flow of the heat-conducting liquid. The linkage component includes an elastic connector and a shelf suspended within the production tank via the elastic connector. A flexible bag containing the IVT reaction solution is disposed on the shelf. During mRNA preparation, the solution agitation component causes the heat-conducting liquid within the production tank to flow. The kinetic energy generated by the flow of the heat-conducting liquid indirectly acts on the flexible reaction bag through the shelf, causing it to sway. The elastic connector connected to the shelf generates a reaction force when subjected to the combined forces of the heat-conducting liquid and the shelf, altering the swaying state of the shelf. This optimizes the force exerted by the shelf on the flexible reaction bag, ensuring that the swaying of the flexible reaction bag maintains the integrity of the mRNA while achieving thorough mixing of the reaction solution. This effectively resolves the contradiction between reaction system uniformity and mRNA structural integrity present in existing IVT reaction scale-up methods, facilitating large-scale industrial production of mRNA and demonstrating promising application prospects. Attached Figure Description
[0017] Figure 1 This is a schematic diagram of the structure of the mRNA production apparatus provided in the first embodiment of this utility model;
[0018] Figure 2 for Figure 1 Sectional view of AA;
[0019] Figure 3 This is a schematic diagram of the mRNA production apparatus provided in the second embodiment of the present invention;
[0020] Figure 4 for Figure 3 A cross-sectional view of BB.
[0021] Reference numerals: 1. Production tank; 11. Tank body; 12. Top cover; 2. First linkage assembly; 21. Spring; 22. First placement plate; 3. Flexible reaction bag; 4. Magnetic stirring assembly; 41. Magnetic stirring table; 42. Magnetic rotor; 5. Temperature controller; 6. Second linkage assembly; 61. U-shaped spring plate group; 62. Second placement plate; 7. Mechanical stirring assembly; 71. Stirring rod; 72. Stirring motor. Detailed Implementation
[0022] Although the present invention can be readily embodied in various forms, only some specific embodiments are shown in the accompanying drawings and will be described in detail in this specification. It is understood that this specification should be regarded as an exemplary illustration of the principles of the present invention and is not intended to limit the present invention to what is described herein.
[0023] Therefore, a feature pointed out in this specification is used to describe one feature of one embodiment of the present invention, and does not imply that every embodiment of the present invention must have the described feature. Furthermore, it should be noted that this specification describes many features. Although certain features may be combined to illustrate possible system designs, these features may also be used in other combinations not explicitly stated. Therefore, unless otherwise stated, the described combinations are not intended to be limiting.
[0024] In the embodiments shown in the accompanying drawings, the directional indications (such as up, down, left, right, front, and back) used to explain the structure and movement of the various elements of this invention are relative rather than absolute. These descriptions are appropriate when these elements are in the positions shown in the drawings. If the descriptions of the positions of these elements change, these directional indications also change accordingly.
[0025] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention.
[0026] Please see Figure 1 and Figure 2 . Figure 1 This is a schematic diagram of the structure of the mRNA production device according to the first embodiment of this utility model. Figure 2 yes Figure 1 A cross-sectional view of AA. The mRNA production apparatus provided in the first embodiment includes: a production tank 1, a first linkage assembly 2, a flexible reaction bag 3, and a magnetic stirring assembly 4.
[0027] The production tank 1 includes a tank body 11 and a top cover 12. The tank body 11 contains a heat-conducting liquid. The first linkage assembly 2 includes a spring 21 and a first placement plate 22. One end of the spring 21 is fixedly connected to the inner wall of the top cover 12, and the other end is fixedly connected to a clamp. The clamp is used to clamp and fix the first placement plate 22, so that the first placement plate 22 is immersed in the heat-conducting liquid in the tank body 11 and kept in a suspended state, and the spring 21 is in a stretched state at this time. The flexible reaction bag 3 is fixed relative to the upper surface of the first placement plate 22 by a fixing rope, so that the flexible reaction bag 3 can shake with the shaking of the first placement plate 22.
[0028] The magnetic stirring assembly 4 includes a magnetic stirring table 41 and a magnetic rotor 42. The production tank 1 is placed on the magnetic stirring table 41, and the magnetic rotor 42 is placed at the bottom of the production tank 1. The magnetic rotor 42 stirs the heat transfer liquid under the drive of the magnetic stirring table 41 to make it flow. During the rotation, the magnetic rotor 42 remains in a non-contact state with the flexible reaction bag 3 and the first placement plate 22.
[0029] A temperature controller 5 is also installed inside the tank 11. The temperature controller 5 is used to detect and control the temperature of the heat transfer fluid inside the tank 11 so that the reaction temperature is kept relatively constant during the IVT reaction, so as to achieve better mRNA preparation effect.
[0030] In this invention, the number of springs 21 in the first linkage component 2 can be adaptively selected according to the placement state of the first shelf 22. The specific number can be 1, 2, 3, 4, or any other integer value greater than these. Furthermore, those skilled in the art can adaptively select the number of springs 21, their connection method with the first shelf 22, and their fixed position as needed. This invention does not impose any particular limitations on these aspects. That is, Figure 2 The number of springs 21, the connection method with the first placement plate 22, and the connection position shown are only one example (first embodiment) of the mRNA production device described in this utility model, but should not be construed as a limitation on the scope of protection of this utility model.
[0031] In this invention, the first placement plate 22 is limited to the placement and fixing of the flexible reaction bag 3, and its specific shape and structure are not particularly limited. In this first embodiment, the first placement plate 22 is preferably a mesh plate, which effectively increases the contact area between the flexible reaction bag 3 and the heat-conducting liquid while effectively fixing the flexible reaction bag 3.
[0032] In this invention, the first placement plate 22 is kept suspended within the tank 11 to allow it to sway with the flow of the heat-conducting liquid within the tank 11. Specifically, it can be placed horizontally, vertically, or tilted within the tank 11. In this first embodiment, the first placement plate 22 is preferably tilted within the tank 11, so that the flexible reaction bag 3 is also tilted. This further optimizes the force acting on the flexible reaction bag 3, achieving good mixing of the reaction liquid within the flexible reaction bag 3 while reducing damage to the mRNA.
[0033] In this utility model, the relative fixing method of the flexible reaction bag 3 and the first placement plate 22 shown in the first embodiment is only an example. Those skilled in the art can select a suitable fixing method from the prior art to achieve the fixing between the flexible reaction bag 3 and the first placement plate 22 according to actual needs. This utility model does not impose any special limitations on it.
[0034] In this invention, the heat-conducting liquid contained in the tank 11 is a type of liquid with heat transfer function commonly used in the prior art, and it is not corrosive to the various components in the mRNA production device. Those skilled in the art can make adaptive selections according to actual usage needs, and this invention does not impose any particular limitations on it. Specific examples can be, but are not limited to, water and / or heat-conducting oil. Specifically, the heat-conducting liquid contained in the tank 11 provided in the first embodiment is water.
[0035] In this invention, the temperature controller 5 is used to monitor and regulate the temperature of the heat transfer fluid inside the tank 11. It is a type of device with temperature detection and control that is commonly used in the prior art. Those skilled in the art can make an adaptive selection according to the actual needs of use. This invention does not impose any special limitations on it.
[0036] In the first embodiment, when using the mRNA production device for amplification IVT reaction of mRNA, the raw materials required for the reaction are added to the flexible reaction bag 3 and fixed on the first placement plate 22. The first placement plate 22 is suspended in the tank 11 by clips on the spring 21, and an appropriate amount of water is added to the tank 11 to completely submerge the first placement plate 22 and the flexible reaction bag 3. Then, the magnetic stirring table 41 is turned on, so that the magnetic rotor 42 in the tank 11 rotates at a speed of 5-200 rpm, thereby agitating the water flow. The internal shear force generated by the water flow acts on the first placement plate 22, causing it to shake. The elasticity of the spring 21 in the stretched state can play a buffering role, optimizing the shaking state of the first placement plate 22. The transmission action of the first placement plate 22 allows the internal shear force generated by the water flow to indirectly act on the flexible reaction bag 3, making the reaction solution inside it more homogeneous and minimizing damage to the mRNA. At the same time, the rotation of the magnetic rotor 42 also helps to maintain the uniformity of the solution temperature throughout the tank 11.
[0037] Please see Figure 3 and Figure 4 . Figure 3 This is a schematic diagram of the structure of the mRNA production device according to the second embodiment of this utility model. Figure 4 yes Figure 3 A BB cross-sectional view. The mRNA production apparatus provided in the second embodiment includes: a production tank 1, a second linkage assembly 6, a flexible reaction bag 3, and a mechanical stirring assembly 7.
[0038] The production tank 1 includes a tank body 11 and a top cover 12. A temperature controller 5 is installed inside the tank body 11, which contains a heat-conducting liquid. The second linkage assembly 6 includes a U-shaped spring plate group 61 and a second placement plate 62. The U-shaped spring plate group 61 is formed by multiple U-shaped springs connected in series, with one side placed at the bottom of the tank body 11 and the other side fixedly connected to the second placement plate 62. This allows the second placement plate 62 to be immersed in the heat-conducting liquid inside the tank body 11 and remain suspended, while the U-shaped spring plate group 61 is in a compressed state. The flexible reaction bag 3 is fixed relative to the lower surface of the second placement plate 62 by a fixing rope, allowing the flexible reaction bag 3 to sway with the movement of the second placement plate 62.
[0039] The mechanical stirring assembly 7 includes a stirring rod 71 and a stirring motor 72. The stirring rod 71 passes through the top cover 12, and the stirring head of the stirring rod 71 is immersed in the heat-conducting liquid, and is in a non-contact state with the flexible reaction bag 3 and the second placement plate 62.
[0040] In the second embodiment, when using the mRNA production device for amplification IVT reaction of mRNA, the raw materials required for the reaction are added to the flexible reaction bag 3 and fixed on the second placement plate 62. The second placement plate 62 is suspended in the tank 11 by a U-shaped spring sheet assembly 61, and an appropriate amount of water is added to the tank 11 to completely submerge the second placement plate 62 and the flexible reaction bag 3. Then, the stirring motor 72 is turned on, so that the stirring rod 71 rotates at a speed of 5-200 rpm, thereby agitating the water flow. The internal shear force generated by the water flow acts on the second placement plate 62, causing it to shake. The elasticity of the U-shaped spring sheet assembly 61 in the compressed state can play a buffering role, optimizing the shaking state of the second placement plate 62. The transmission action of the second placement plate 62 allows the internal shear force generated by the water flow to indirectly act on the flexible reaction bag 3, making the reaction solution inside mixed evenly and minimizing damage to the mRNA. At the same time, the rotation of the stirring rod 71 also helps to maintain the uniformity of the solution temperature throughout the tank 11.
[0041] mRNA was prepared using the mRNA production apparatus provided in the first and second embodiments. The total volume of the reaction solution was 20 mL, and the specific amounts of each reaction raw material and the reaction conditions are shown in Table 1.
[0042] Table 1.
[0043]
[0044] The DNA template includes nucleotide fragments with sequences such as SEQ ID NO:1, as shown in Table 2.
[0045] Table 2.
[0046]
[0047]
[0048] Take 50 μL of the final IVT reaction solution, and use the RNA purification kit (NEB, catalog number T2050L) according to the instructions to obtain the mRNA purification product. Then, use the RNA mode of the Nanodrop micro spectrophotometer to detect the total amount of mRNA in the IVT reaction solution (in μg), and calculate the mRNA yield (mg / mL) of 50 μL of IVT reaction solution provided in each example according to the following formula.
[0049] mRNA yield (mg / mL) = X * 21 / 1000
[0050] X represents the total mRNA obtained after purification from a 50 μL sample of IVT reaction solution. The results are shown in Table 3.
[0051] Table 3.
[0052] Group mRNA yield (mg / mL) Example 1 4.9 Example 2 5.1
[0053] As shown in Table 3, the test results indicate that the mRNA production devices provided in Examples 1 and 2 of this invention can effectively solve the contradiction between the uniformity of the reaction system and the integrity of the mRNA structure in the existing IVT reaction amplification. The mRNA yields are 4.9 mg / mL and 5.1 mg / mL, respectively, achieving milligram-level preparation of mRNA. This is beneficial for the industrial-scale amplification of mRNA production and has good application prospects.
[0054] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention without departing from the principles and spirit of the present invention.
Claims
1. An mRNA production apparatus, characterized by, The mRNA production device comprises: a production tank, a heat-conducting liquid is arranged in the production tank; a linkage assembly is arranged in the production tank, the linkage assembly comprises an elastic connecting piece and a placing plate, the elastic connecting piece is connected with the peripheral wall of the production tank, and the placing plate is connected with the elastic connecting piece to be suspended in the production tank; the placing plate is a grid plate, and the placing plate is arranged in an inclined manner; a flexible reaction bag is arranged on the placing plate, and the flexible reaction bag and the placing plate are soaked in the heat-conducting liquid; a solution disturbance assembly is used to drive the heat-conducting liquid to flow to shake the placing plate, and the solution disturbance assembly keeps a non-contact state with the flexible reaction bag and the placing plate; wherein the linkage assembly comprises one or more of the following features: (1) the production tank comprises a tank body and a top cover; one end of the elastic connecting piece is connected with the top cover, and the other end of the elastic connecting piece is connected with the placing plate, so that the elastic connecting piece is in a tensile state; (2) the bottom wall of the production tank is connected with one end of the elastic connecting piece, and the other end of the elastic connecting piece is connected with the placing plate, so that the elastic connecting piece is in a compression state.
2. The mRNA production device of claim 1, wherein, The solution disturbance assembly is a magnetic stirring assembly, the magnetic stirring assembly comprises a magnetic stirring table and a magnetic rotor, the production tank is arranged on the magnetic stirring table, the magnetic rotor is arranged in the production tank to drive the heat-conducting liquid to flow, and the magnetic rotor keeps a non-contact state with the flexible reaction bag and the placing plate.
3. The mRNA production device of claim 2, wherein, The flexible reaction bag is arranged on the side of the placing plate away from the magnetic rotor.
4. The mRNA production device of claim 1, wherein, The solution disturbance assembly is a mechanical stirring assembly, the mechanical stirring assembly comprises a stirring rod and a stirring motor, the stirring motor drives the stirring rod to rotate, the stirring rod is inserted into the production tank to drive the heat-conducting liquid to flow, and the stirring rod keeps a non-contact state with the flexible reaction bag and the placing plate.
5. The mRNA production device of claim 4, wherein, The flexible reaction bag is arranged on the side of the placing plate away from the stirring rod.
6. The mRNA production device of claim 1, wherein, A temperature controller is arranged in the production tank.