High throughput lnp preparation system

Abstract

The utility model provides high -flux LNP preparation system, including water -phase flow path for LNP package, organic phase flow path for LNP package, mixer group, aftereffect flow path, water -phase flow path includes water -phase sample feeder, water -phase sample feed switch valve, and water -phase sample feeder is used for storing and selecting water -phase sample, water -phase sample feed switch valve and water -phase sample feeder intercommunication, water -phase flow path still includes water -phase pump group, and water -phase pump group is linked with water -phase sample feed switch valve intercommunication. The utility model discloses the automatic sample feeding of water -phase / organic phase, multiple different volume constant -flow injection pump, multiple different micro -fluidic chip arbitrary selection and automatic switching, on -line dilution and automatic collection etc. Key steps are integrated in the same automation platform. With control system, can realize the automatic operation preparation LNP of serialization, including the automatic cleaning of system, greatly reduces the manual intervention, reduces the pollution risk, can realize the high efficiency, high -flux preparation of LNP.

Description

Technical Field

[0001] This utility model relates to the field of high-throughput LNP preparation, specifically a high-throughput LNP preparation system. Background Technology

[0002] mRNA-LNPs have shown great potential in both preventative and therapeutic vaccines. However, mRNA is prone to degradation in vitro, has a short half-life in vivo, low intracellular conversion rate (due to its charged nature), and activates the innate immune system, resulting in poor drug-likeness. Therefore, LNPs are indispensable delivery systems. LNPs can protect RNA from degradation by physicochemical and enzymatic factors, increase its intracellular delivery and endosome escape efficiency, and inhibit the host immune system's recognition of exogenous mRNA. With the expansion of mRNA-LNP applications, such as different indications and routes of administration, there is a need for sequence optimization and modification of mRNA, screening and establishing ionizable lipid libraries, screening the liposome composition and ratio of LNPs, and optimizing and screening LNP preparation processes. All of these are essential factors determining speed and success. However, these essential factors involve a significant amount of time-consuming and labor-intensive work and are prone to errors. Therefore, there is an urgent need for automated high-throughput mRNA-LNP preparation systems.

[0003] Currently, research and development or process development still relies on manual methods such as changing syringes and cleaning chips, resulting in low efficiency in preparing or screening LNPs. Some companies use automated pipetting workstations for high-throughput preparation of mRNA-LNPs, which can greatly improve the efficiency of mRNA-LNP preparation. However, automated pipetting workstations use the method of blowing liquid to promote the mixing of oil and water phases rather than a microfluidic mode. This can lead to differences in the size, morphology, and internal structure of LNPs, which in turn affect their efficacy and toxicity. This makes it impossible to bridge to industrial production, rendering the high-throughput effort useless. There are also high-throughput mRNA-LNP systems based on microfluidics, such as Sunscreen. However, the power of the oil-water phase fluid in this system comes from the agitation of the incompatible liquid, which poses risks, high screening costs, and environmental pollution problems. Utility Model Content

[0004] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a high-throughput LNP preparation system. This system, through a highly integrated automated design, solves problems such as low efficiency in large-scale mRNA-LNP screening and the inability to directly transform or bridge to scale-up and production.

[0005] Technical solution: To achieve the above objectives, this utility model provides a high-throughput LNP preparation system, including an aqueous phase flow path for LNP encapsulation, an organic phase flow path for LNP encapsulation, a mixer assembly for oil-water phase mixing to form LNP, and a post-effect flow path and collection valve for LNP dilution, LNP pre-disposal or collection. The aqueous phase flow path includes an aqueous phase injector and an aqueous phase injection switching valve. The aqueous phase injector is used to store and select aqueous phase samples. The aqueous phase injection switching valve is connected to the aqueous phase injector. The aqueous phase flow path also includes an aqueous phase pump set, which is connected to the aqueous phase injection switching valve. The organic phase flow path includes an organic phase injector and an organic phase injection switching valve. The organic phase injector is used to store and select organic phase samples. The organic phase injection switching valve is connected to the organic phase injector. The organic phase flow path also includes an organic phase pump assembly, which is connected to the organic phase injection switching valve. The mixer assembly is connected to both the aqueous phase pump assembly and the organic phase pump assembly; Preferably, the aqueous flow path injector includes a variety of aqueous sample containers.

[0006] Preferably, the organic phase flow path injector includes a variety of organic phase sample containers.

[0007] Preferably, the aqueous phase pump set includes a low-volume aqueous phase pump, a medium-volume aqueous phase pump, and a high-volume aqueous phase pump.

[0008] Preferably, the organic phase pump group includes a low-volume organic phase pump, a medium-volume organic phase pump, and a high-volume organic phase pump.

[0009] Preferred, The pump set is a continuous injection pump, which can be selected for continuous injection or single injection; The injection volume of the low-volume aqueous phase pump is 5 ml; The injection volume of the medium-volume aqueous phase pump is 10 ml; The injection volume of the high-volume aqueous phase pump is 25 ml.

[0010] Preferred, The pump set is a continuous injection pump, which can be selected for continuous injection or single injection; The injection volume of the low-volume organic phase pump is 1 ml; The injection volume of the medium-volume organic phase pump is 5 ml; The injection volume of the high-volume organic phase pump is 10 ml.

[0011] Preferably, the mixer group includes multiple mixers with different structural dimensions, and each mixer is connected to a chip selection valve.

[0012] Preferably, the aftereffect flow path further includes a collector and a dilution pump, characterized in that the LNP can be continuously collected or collected in separate pipes according to time and volume; Preferably, the aqueous phase flow path further includes an aqueous phase waste liquid flow channel; the organic phase flow path further includes an organic phase waste liquid flow channel.

[0013] Compared with the prior art, the present invention has the following significant advantages: 1. High Flexibility and Adaptability: This invention, by setting up multiple specifications (low, medium, and high volume) of aqueous phase pump sets and organic phase pump sets, combined with corresponding multi-specification mixer sets, allows the system to flexibly select different pump and mixer combinations according to needs. For example, it can achieve the preparation of LNPs with different total volumes such as 1-4ml, 4-10ml to 10-33ml through single injection mode, and can also achieve unlimited volume preparation through continuous injection mode. This design greatly improves the adaptability to batch scale, meeting the needs from formulation screening in the R&D stage to process scale-up research.

[0014] 2. Precise proportion control: The system uses parallel operation of the aqueous phase pump group and the organic phase pump group, and precisely controls the flow ratio of the two, thereby achieving flexible and high-precision control of the LNP preparation proportion (such as N / P ratio).

[0015] 3. High degree of automation and integration: This invention integrates key steps such as automatic sampling of aqueous / organic phases, multi-channel pumping, multi-stage mixing, online dilution, and automatic collection into a single automated platform. Combined with a control system, it enables sequential automatic operation, including automatic system cleaning, greatly reducing manual intervention and lowering the risk of contamination and errors. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 This is a pipeline diagram of one embodiment of the present invention.

[0018] Explanation of reference numerals in the attached figures: 1. Aqueous phase flow path; 11. Aqueous phase injector; 12. Aqueous phase injection switching valve; 13. Aqueous phase pump set; 131. Low volume aqueous phase pump; 132. Medium volume aqueous phase pump; 133. High volume aqueous phase pump; 2. Organic phase flow path; 21. Organic phase injector; 22. Organic phase injection switching valve; 23. Organic phase pump assembly; 231. Low-volume organic phase pump; 232. Medium-volume organic phase pump; 233. High-volume organic phase pump; 3. Aftereffect flow path; 31. Mixer assembly; 32. Dilution pump; 33. Collector; 34. Mixer outlet selector valve. Detailed Implementation

[0019] The accompanying drawings are for illustrative purposes only and should not be construed as limiting the scope of this patent. To better illustrate this embodiment, some parts in the drawings may be omitted, enlarged, or reduced, and do not represent the actual dimensions of the product.

[0020] It will be understood by those skilled in the art that certain well-known structures and their descriptions may be omitted in the accompanying drawings. The technical solution of this utility model will be further described below with reference to the accompanying drawings and embodiments.

[0021] The high-throughput LNP preparation system of this invention mainly includes an aqueous phase flow path 1, an organic phase flow path 2, an aftereffect flow path 3, and a mixer group 31.

[0022] The aqueous phase flow path 1 is used for the precise delivery of the aqueous phase solution (such as a buffer for mRNA) required for LNP encapsulation. This flow path includes an aqueous phase injector 11, an aqueous phase injection switching valve 12, and an aqueous phase pump assembly 13.

[0023] The aqueous phase injector 11 is used for storing and automatically selecting aqueous phase samples. In one embodiment, the aqueous phase injector 11 is compatible with various sample rack sizes, such as racks for ≥72 4ml sample vials, racks for ≥24 10ml sample vials and ≥24 20ml sample vials, as well as racks for 48-well plates and 4 24-well plates.

[0024] The aqueous phase injection switching valve 12 is used to select the source of the aqueous phase. The aqueous phase injection switching valve 12 can be connected to the aqueous phase injector 11, or to multiple large-volume aqueous phase storage bottles and cleaning / purging lines.

[0025] The aqueous phase pump assembly 13 is connected to the aqueous phase injection switching valve 12. To accommodate different batch (scale) LNP preparation requirements, the pump assembly is preferably composed of multiple continuous injection pumps of different volumes. In one embodiment, the aqueous phase pump assembly 13 includes: Single injection mode: Low-volume aqueous phase pump 131: for example, a 1 ml syringe pump for preparation of 1-4 ml total volume.

[0026] Medium volume aqueous phase pump 132: for example, a 5ml syringe pump for preparing 4-10ml total volume.

[0027] High-volume aqueous phase pump 133: e.g., a 10ml syringe pump for preparing 10-33ml total volume.

[0028] Organic phase flow path 2 is used for precise delivery of the organic phase solution (such as an ethanol solution containing dissolved lipids) required for LNP encapsulation. This flow path structure corresponds to the aqueous phase flow path 1 in mirror and includes an organic phase injector 21, an organic phase injection switching valve 22, and an organic phase pump assembly 23.

[0029] Organic phase injector 21 is used for storing and automatically selecting organic phase samples. In one embodiment, injector 2 is compatible with, for example, a holder for ≥120 2ml sample vials, ≥72 4ml and ≥24 10ml sample vials, as well as four 96-well plates and four 48-well plates.

[0030] The organic phase injection switching valve 22 is used to select the source of the organic phase. It can be connected to the injector 2, or to multiple large-volume organic phase storage bottles and cleaning / purging lines.

[0031] The organic phase pump assembly 23 is connected to the organic phase injection switching valve 22. Similarly, the organic phase pump assembly 23 is preferably composed of multiple continuous injection pumps of different volumes. In one embodiment, the organic phase pump assembly 23 includes a low-volume organic phase pump 231, a medium-volume organic phase pump 232, and a high-volume organic phase pump 233.

[0032] The aftereffect flow path 3 is used for the preparation of LNP by mixing organic and aqueous phases, as well as the online dilution and collection of LNP. This flow path includes a mixer assembly 31, a dilution pump 32, and a collector 34.

[0033] The mixer assembly 31 is connected to both the aqueous phase pump assembly 13 and the organic phase pump assembly 23. Aqueous fluid enters the mixer assembly 31 via an aqueous phase output switching valve; organic fluid enters the mixer assembly 31 via an organic phase output switching valve. System control valves ensure that both aqueous and organic phases select the same mixer at the same time. Preferably, the mixer assembly 31 is also configured with low, medium, and high specifications to match pump assemblies of different volumes, ensuring efficient mixing at different flow rates and batch sizes.

[0034] The mixer outlet is connected to mixer outlet selector valve 33, which introduces the initial LNP solution downstream.

[0035] Downstream, the flow path is selected via valves: one path can discharge waste; one path can directly enter collector 34 via valves; and the other path enters the online dilution pipeline.

[0036] The dilution pump 32 is used to pump the dilution buffer. After the initial LNP solution is mixed with the dilution buffer, it flows through a valve, where it can be either discharged as waste or enters the collector 34 for automatic continuous or stepwise collection.

[0037] Collector 34 is preferably compatible with a variety of collection tubes, such as ≥42 50ml centrifuge tubes and ≥120 15ml centrifuge tubes, as well as ≥4 48-well plates and ≥4 24-well plates.

[0038] In actual operation, the system prepares batches and formulations based on preset LNPs, and executes them automatically through the control system: Select Batch Size: Choose between the injection pump group and single injection mode for automatic preparation of LNPs in different small volumes (with varying flow rates and corresponding precision). For example, when performing small-batch screening (e.g., 1-4 ml), select the low-volume aqueous phase pump 131 and the low-volume organic phase pump 231. Alternatively, a continuous injection mode can be selected to achieve unlimited volume preparation.

[0039] LNP preparation (ratio control): The aqueous phase pump group 13 and the organic phase pump 23 operate in parallel (constant-rate injection), pushing the two-phase solutions into one of the selected mixer groups 31 according to the preset flow rate ratio for mixing to form LNP. By precisely controlling the flow rate ratio of the two pumps, the LNP preparation ratio can be precisely controlled.

[0040] In some embodiments, the output port of the aqueous phase pump group 13 is also connected to an aqueous phase output mixer switching valve, which is used to select the output of the aqueous phase pump group 13 to be input to one of the mixers in the mixer group 31. The output port of the organic phase pump group 23 is also connected to an organic phase output mixer switching valve, which is used to select the output of the organic phase pump group 23 to be input to a certain mixer in the mixer group 31.

[0041] Dilution and collection: The generated initial LNP liquid flows through the mixer outlet selection valve 33 and is mixed with the diluent from the dilution pump 32. Then it enters the collector 34 through the valve pipeline and is automatically collected to the designated collection tube or orifice plate position according to the preset volume or time.

[0042] Automatic cleaning: After a formulation is prepared, the system can automatically switch to cleaning solution (water, ethanol, alkali) and gas line to automatically clean and purge the injection needle, tubing, pump and mixer.

[0043] Through the aforementioned highly automated integrated design, this system achieves full automation of the entire process, from sample injection, multi-batch LNP preparation, online dilution to automatic collection and system cleaning, perfectly meeting the flexibility, automation, and continuity requirements of high-throughput LNP formulation screening and process development.

[0044] Obviously, the above embodiments of this utility model are merely examples for clearly illustrating this utility model, and are not intended to limit the implementation of this utility model. Those skilled in the art can make other variations or modifications based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this utility model should be included within the protection scope of the claims of this utility model.

Claims

1. A high-throughput LNP preparation system, characterized in that, It includes an aqueous phase flow path for LNP encapsulation, an organic phase flow path for LNP encapsulation, a mixer assembly for mixing oil and water phases to form LNP, and an aftereffect flow path and collection valve for LNP dilution, LNP pre-disposal or collection. The aqueous phase flow path includes an aqueous phase injector and an aqueous phase injection switching valve. The aqueous phase injector is used to store and select aqueous phase samples. The aqueous phase injection switching valve is connected to the aqueous phase injector. The aqueous phase flow path also includes an aqueous phase pump set, which is connected to the aqueous phase injection switching valve. The organic phase flow path includes an organic phase injector and an organic phase injection switching valve. The organic phase injector is used to store and select organic phase samples. The organic phase injection switching valve is connected to the organic phase injector. The organic phase flow path also includes an organic phase pump assembly, which is connected to the organic phase injection switching valve. The mixer assembly is connected to both the aqueous phase pump assembly and the organic phase pump assembly.

2. The high-throughput LNP preparation system according to claim 1, characterized in that, Aqueous flow path samplers include a variety of aqueous sample containers.

3. The high-throughput LNP preparation system according to claim 1, characterized in that, Organic phase flow path injectors include a variety of organic phase sample containers.

4. The high-throughput LNP preparation system according to claim 1, characterized in that, The aqueous phase pump set includes a low-volume aqueous phase pump, a medium-volume aqueous phase pump, and a high-volume aqueous phase pump.

5. The high-throughput LNP preparation system according to claim 1, characterized in that, The organic phase pump group includes a low-volume organic phase pump, a medium-volume organic phase pump, and a high-volume organic phase pump.

6. The high-throughput LNP preparation system according to claim 4, characterized in that, The pump set is a continuous injection pump, which can be selected for continuous injection or single injection; The injection volume of the low-volume aqueous phase pump is 5 ml; The injection volume of the medium-volume aqueous phase pump is 10 ml; The injection volume of the high-volume aqueous phase pump is 25 ml.

7. The high-throughput LNP preparation system according to claim 5, characterized in that, The pump set is a continuous injection pump, which can be selected for continuous injection or single injection; The injection volume of the low-volume organic phase pump is 1 ml; The injection volume of the medium-volume organic phase pump is 5 ml; The injection volume of the high-volume organic phase pump is 10 ml.

8. The high-throughput LNP preparation system according to claim 1, characterized in that, The mixer group includes multiple mixers with different structural dimensions, and each mixer is connected to a chip selection valve.

9. The high-throughput LNP preparation system according to claim 1, characterized in that, The aftereffect flow path also includes a collector and a dilution pump. The collector can collect LNP continuously or in separate pipes according to time and volume.

10. The high-throughput LNP preparation system according to claim 1, characterized in that, The aqueous phase flow path also includes an aqueous phase waste liquid flow path; the organic phase flow path also includes an organic phase waste liquid flow path.

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