Micro-fluidic chip for synthesizing mRNA (messenger ribonucleic acid) lipid nanoparticles
By employing a double-sided herringbone structure in a microfluidic chip, the problem of uneven mixing in a single-sided mixing system was solved, enabling more efficient and uniform synthesis of lipid nanoparticles, thereby improving production efficiency and product quality.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-24
- Publication Date
- 2026-03-10
AI Technical Summary
Existing micro-hybrid chips are single-sided mixing systems, resulting in uneven mixing and unstable parameters, leading to poor mixing effects and impacting production and economic efficiency.
The microfluidic chip design employs a dual-sided herringbone structure. Through the synergistic effect of the top and bottom templates, the fluid is promoted to be divided and merged multiple times within the chip, resulting in thorough mixing and optimized fluid control accuracy.
This improved mixing quality and consistency, reduced reagent costs, increased production efficiency, and synthesized lipid nanoparticles with narrower particle size distribution and better encapsulation effect.
Smart Images
Figure CN223980513U_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The utility model relates to microfluidics and microreactor technical field, concretely to a kind of microfluidic chip for synthesizing mRNA lipid nanoparticle. BACKGROUND
[0002] Messenger RNA vaccines represent a revolutionary approach to infectious disease prevention, bringing great prospects for cancer immunotherapy, since the COVID-19 outbreak and the success of two mRNA vaccines against SARS-COV-2, mRNA technology has rapidly become the focus of vaccine development;
[0003] Although early studies showed that naked mRNA could be directly injected without carriers to express proteins in mouse muscle cells, the inherent instability of mRNA molecules, susceptibility to RNase degradation and negatively charged properties limit their cellular uptake and clinical potential, lipid-based delivery systems and polymer-based delivery systems are two important mRNA delivery platforms, with the emergence of efficient mRNA delivery vectors, especially the success of COVID-19 vaccine delivery systems, lipid nanoparticles containing ionizable cationic lipids have completely changed mRNA therapy;
[0004] The existing preparation methods of LNP mainly include traditional Y-shaped interface and T-shaped interface collision flow design, collision flow design with special structure optimization, separation recombination structure microchannel mixing technology represented by Tesla structure and microchannel mixing technology with embedded structure disturbance increase, there are still many deficiencies as follows:
[0005] Most are single-sided mixing systems, and after intersection, preliminary mixing may only be carried out in local area, which is easy to mix unevenly, and the mixing effect is poor, and the parameters of the chip and its internal structures are unstable, the precision of fluid control is low, which easily leads to poor solution collision effect, affects the overall mixing quality, and the production benefit and economic benefit are poor. UTILITY MODEL CONTENTS
[0006] The utility model aims at providing a kind of microfluidic chip for synthesizing mRNA lipid nanoparticle to solve the problems of the current herringbone micro-mixing chip in the above background technology, which is mostly single-sided mixing system, and after intersection, preliminary mixing may only be carried out in local area, which is easy to mix unevenly, and the mixing effect is poor, and the parameters of the chip and its internal structures are unstable, the precision of fluid control is low, which easily leads to poor solution collision effect, affects the overall mixing quality, and the production benefit and economic benefit are poor.
[0007] In order to achieve the above object, the utility model provides the following technical scheme: A micro -fluidic chip for synthesis mRNA lipid nano -particle, including chip main part, the top of chip main part Fixedly be equipped with top layer template, the bottom of chip main part Fixedly be equipped with bottom layer template, top layer template bears the function of solution introduction and other interactions with the outside world in subsequent use, the structure design of through its own and chip main part, bottom layer template cooperate to influence the solution in the chip's direction, bottom layer template firm the structure of whole chip main part on one side, on the other hand, with top layer template cooperates to guide the mixing of fluid, reaction process etc., promotes the synthesis of bottom layer template mRNA bottom layer template lipid nano -particle.
[0008] Preferably, the bottom surface of the top layer template and the surface of the bottom layer template are provided with a plurality of uniformly distributed fishbone-shaped grooves, the surfaces of the plurality of fishbone-shaped grooves are fixedly connected with herringbone fish; the unique shape of the herringbone fish makes the fluid be divided and merged when flowing, the fluid flowing to the branch of the herringbone fish will be divided into different small streams, and then merged together in the subsequent flowing process, the process is repeated, which is like "stirring" the solution multiple times, so that the mRNA solution and the lipid solution can be more fully mixed, greatly improving the overall mixing performance, the top layer template and the bottom layer template are provided with herringbone fish structures on both sides, compared with single-sided setting, the double-sided setting doubles the promotion effect of fluid mixing, making the mixing in the whole chip more uniform and comprehensive, avoiding the problems of uneven mixing and large local differences that may occur in single-sided structure, and helping to synthesize mRNA lipid nanoparticles with more stable quality and better consistency.
[0009] Preferably, the peak side angle of the herringbone fishbone is 45°, the height of the herringbone fishbone is 30μm, and the spacing between the two herringbone fishbones is 50μm; when the fluid encounters the herringbone fishbone, it will be naturally divided into two relatively uniform tributaries according to the angle of the 45° bottom template, neither too violent disturbance of the fluid due to the angle, leading to fluid flow disorder, affecting the accuracy of mixing, nor the inability to fully disrupt the original flow state of the fluid due to the angle being too small, ensuring that the solutions can effectively interpenetrate and mix, which helps to improve the overall mixing efficiency. When the fluid flows through the herringbone fishbone, the protruding bottom template with a height of 30μm will cause the fluid to change the flow direction and exchange different layers of fluid, etc., prompting different parts of different solutions to mix with each other, thereby strengthening the entire mixing process. At the same time, the height of the 30μm bottom template is not too high and will not cause excessive hindrance to the normal flow of the fluid. On the premise of ensuring effective promotion of mixing, the fluid flows relatively smoothly, ensuring efficient synthesis process, and the 50μm spacing between the two herringbone fishbones can well control the rhythm of fluid mixing and provide a suitable bottom template "buffer" space for the fluid, allowing the fluid that has undergone preliminary mixing to further adjust itself and interact with the surrounding fluid in this space, and then enter the next herringbone fishbone structure for a new round of mixing, making the mixing more thorough and orderly, and avoiding the influence of too frequent or too sparse mixing on the final synthesis effect.
[0010] Preferably, the total height of the chip body is 80μm, and the height of the herringbone fishbone is 30μm. In the overall space of the 80μm bottom template, the fishbone structure has enough "space to play" to effectively disturb the fluid and guide the fluid mixing, and the space is not too cramped, so that the fluid can continue to flow in the expected direction and state when passing through the fishbone structure, avoiding fluid disorder or blockage that is not conducive to mixing due to insufficient space.
[0011] Preferably, the top layer template is internally provided with two sample inlets, and the top layer template is internally provided with a product outlet, the two sample inlets can correspondingly introduce the two different solutions respectively, through different sample inlets, the starting time, flow rate and other parameters of each solution entering the chip can be accurately controlled, so that the subsequent precise and orderly mixing in the chip is created, compared with single inlet or multiple dispersed inlets, the double-side sampling mode is more conducive to realizing the rapid and uniform mixing of the solution in the limited space of the microfluidic chip, the product outlet serves as a product output channel, ensuring that the final synthesized product can be collected for subsequent application, the solution enters from the sample inlet, is fully mixed in the chip body by means of the herringbone fishbone structure and completes the synthesis reaction, and finally flows out through the product outlet, the clear and orderly layout is helpful to realize the precise control and efficient operation of the whole synthesis process, and guarantees the quality and yield of the synthesized bottom layer template mRNA bottom layer template lipid nanoparticles, so that it is more in line with the application requirements in the field of biological medicine.
[0012] Preferably, the length of the sample inlet is 20mm, the included angle of the two sample inlets is 120°, and the length of the product outlet is 5mm; the 20mm sample inlet length is helpful to realize a stable fluid introduction process, so that the solution can have a relatively stable bottom layer template "transition area" before entering the chip body, so that the solution can enter the preset mixing channel in the chip body in a more uniform and stable state, laying a good foundation for subsequent mixing operation, when the bottom layer template mRNA bottom layer template solution and the lipid solution enter the chip from the two sample inlets respectively, they will intersect in the chip at an included angle of 120°, such an intersection angle can promote the two solutions to form a suitable collision and interpenetration mode when meeting, which is helpful to realize more balanced diffusion and flow of the solution in the chip, and the length of the product outlet is set to 5mm, which can neither cause the product to accumulate and block during the flow-out process, nor be too short to make the product flow out too fast, so as to ensure the stability of fluid flow in the whole chip and effective collection of the synthesized product.
[0013] Compared with the prior art, the microfluidic chip for synthesizing mRNA lipid nanoparticles has the following beneficial effects:
[0014] The microfluidic chip for synthesizing mRNA lipid nanoparticles uses a small number of herringbone fishbone mixers to form a double-side mixing promotion system, which can reduce the cost of reagents while ensuring the synthesis quality and achieve better mixing effect, and the chip and its internal structure parameters are carefully optimized, which can accurately control the fluid, help to synthesize bottom layer template mRNA bottom layer template lipid nanoparticles with higher quality, narrower particle size distribution and better wrapping effect, indirectly improve the production efficiency, and further embody the advantages of the prior art in cost control and resource utilization. Attached Figure Description
[0015] Figure 1 This is a schematic diagram of the core hybrid module of this utility model;
[0016] Figure 2 This is a schematic diagram of the structure of the mRNA lipid nanoparticles synthesized according to this invention;
[0017] Figure 3 This is a schematic diagram of the structure of the mRNA lipid nanoparticles synthesized according to this invention;
[0018] Figure 4 This is a schematic diagram of the structure of the LNP of this utility model;
[0019] Figure 5 This is a schematic diagram illustrating the mixing effect of the chip described in Embodiment 1 of this utility model;
[0020] Figure 6 This is a schematic diagram of the mixing effect of the chip described in Embodiment 2 of this utility model;
[0021] Figure 7 This is a schematic diagram illustrating the mixing effect of the single-sided control chip used in this invention.
[0022] In the diagram: 1. Chip body; 2. Top layer template; 3. Bottom layer template; 4. Fishbone-shaped groove; 5. Herringbone pattern; 6. Sample inlet; 7. Output outlet. Detailed Implementation
[0023] 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.
[0024] Example 1:
[0025] This embodiment provides a preferred embodiment 1 of a microfluidic chip for synthesizing mRNA lipid nanoparticles. The chip consists of a top template 2 and a bottom template 3 containing a herringbone structure. The herringbone structures 5 within the serpentine channels between the top template 2 and the bottom template 3 are arranged periodically in an alternating pattern, collectively forming a hybrid module of the microfluidic chip. A schematic diagram of this hybrid module is shown below. Figure 1 As shown, the staggered herringbone structure is arranged in cycles of 24, with each cycle containing 12 herringbone structures 5 on the top layer template 2 and 12 herringbone structures 5 on the bottom layer template 3. The 12 herringbone structures 5 on the top layer template 2 and the bottom layer template 3 are each composed of 6 left-side and 6 right-side herringbone structures 5. This embodiment includes 3 hybrid modules, totaling 72 herringbone structures 5. Figure 2As shown above, in addition, two injection ports 6 are located upstream of the mixing module, and the output port 7 is located downstream of the mixing module.
[0026] Please see Figures 1-7 This invention provides a microfluidic chip for synthesizing mRNA lipid nanoparticles, including a chip body 1, a top template 2 fixedly disposed at the upper end of the chip body 1, and a bottom template 3 fixedly disposed at the bottom end of the chip body 1.
[0027] During use, the chip body 1 provides a stable physical support for the top template 2 and the bottom template 3. The top template 2 ensures that the solution can smoothly enter the chip during use and that the product can be collected, and prevents solution leakage. The bottom template 3 bears the weight of the solution inside the chip and the pressure generated, prevents deformation of the bottom of the chip body 1, and ensures that the internal spatial structure and fluid channels of the chip body 1 remain stable.
[0028] Furthermore, the bottom surface of the top template 2 and the surface of the bottom template 3 are provided with several evenly distributed fishbone-shaped grooves 4, and the surfaces of the several fishbone-shaped grooves 4 are fixedly connected with herringbone-shaped fish bones 5.
[0029] During use, the fishbone-shaped groove 4 increases the contact area between the solution and the internal structure of the chip, allowing the fluid to flow towards the herringbone 5, preparing for subsequent mixing operations. The herringbone 5 divides the fluid into different small streams, allowing the mRNA solution and lipid solution to mix more thoroughly.
[0030] Furthermore, the angle between the peaks of the herringbone 5 is 45°, the height of the herringbone 5 is 30μm, and the distance between the two herringbone 5s is 50μm.
[0031] During use, the 45° peak-edge angle allows the fluid to maintain a relatively stable state during splitting and merging. The 30μm height disturbs the fluid, changing its local flow velocity and direction. This allows for more thorough fusion of solutions without affecting the normal flow of the fluid. The 50μm spacing between the two herringbone-shaped sections controls the frequency of fluid splitting and merging, making the mixing process rhythmic and ensuring that the mixing of fluids within the entire chip is uniform and efficient.
[0032] Furthermore, the total height of the chip body 1 is 80μm;
[0033] When in use, the 80μm height of the chip body 1 can well accommodate the fishbone-shaped grooves 4 and herringbone-shaped fishbone 5 on the surfaces of the top template 2 and the bottom template 3.
[0034] Furthermore, the top template 2 has two sample inlets 6 inside and a production outlet 7 inside;
[0035] In use, the two solutions enter the chip body 1 through the two inlets 6 respectively. After completing a series of reactions such as mixing and encapsulation inside the chip, they leave the chip smoothly through the outlet 7 and are collected for subsequent applications.
[0036] Furthermore, the length of the injection port 6 is 20 mm, and the included angle between the two injection ports 6 is 120°, while the length of the output port 7 is 5 mm.
[0037] During use, when the mRNA solution and lipid solution are delivered from the outside to the injection port 6 through the tubing, the 20mm length provides a buffer zone for the solution, allowing it to gradually adapt to the transition from the external tubing to the inside of the chip. The angle between the two injection ports 6 is 120°, allowing the two solutions to collide and intersect appropriately at the moment of convergence, enabling them to start the mixing process quickly and uniformly. The length of the outlet 7 is 5mm, allowing the product to pass through the outlet 7 efficiently and unimpeded along the natural flow direction of the fluid, maintaining the complete process of the entire chip from sample introduction, synthesis to product export.
[0038] Example 2:
[0039] This embodiment provides a preferred embodiment 2 of a microfluidic chip for synthesizing mRNA lipid nanoparticles. The chip consists of a top template 2 and a bottom template 3 containing a herringbone structure. The herringbone structures 5 within the serpentine channels between the top template 2 and the bottom template 3 are arranged periodically in an alternating pattern, collectively forming a hybrid module of the microfluidic chip. A schematic diagram of this hybrid module is shown below. Figure 1 As shown, the staggered herringbone structures are arranged in cycles of 24, with each cycle containing 12 herringbone structures 5 on the top layer template 2 and 12 herringbone structures 5 on the bottom layer template 3. The 12 herringbone structures 5 on the top layer template 2 and the bottom layer template 3 are each composed of 6 left-side and 6 right-side herringbone structures 5. This embodiment includes two hybrid modules, totaling 48 herringbone structures, as shown... Figure 2 As shown above, in addition, two injection ports 6 are located upstream of the mixing module, and the output port 7 is located downstream of the mixing module.
[0040] Example 3:
[0041] This embodiment, as another embodiment of the present invention, provides a method for synthesizing lipid nanoparticles using the microfluidic chip described in Embodiments 1 and 2. First, a lipid-ethanol solution is prepared using a molar ratio of 50% ALC-0315, 1.5% ALC-0159, 10% DSPC, and 38.5% cholesterol. Then, an mRNA-citric acid buffer is prepared by dissolving the mRNA in an acidic citrate buffer at pH 4 and diluting it to the desired concentration according to the N / P ratio and FRR. Next, the lipid phase solution and aqueous phase solution are respectively aspirated using syringes and connected to inlets A and B. The injection rates of the lipid and aqueous phase solutions are adjusted. The mixed liquid is collected at reactant outlet C (discarding the waste liquid before and after 1-2 seconds). The mixed mRNA lipid nanoparticles are ultrafiltered using Tris-HCl buffer. The particle size and PDI of the LNPs are detected using a nanoparticle size and Zeta potential analyzer. Finally, a Quant-iT... TM RNA reagent was used to determine the encapsulation efficiency of LNPs, and the final results showed that the encapsulation efficiency of LNPs was greater than 98%.
[0042] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A microfluidic chip for the synthesis of mRNA lipid nanoparticles comprising a chip body (1), characterized in that: The upper end of the chip body (1) is fixedly provided with a top template (2), and the bottom end of the chip body (1) is fixedly provided with a bottom template (3); The bottom surface of the top template (2) and the surface of the bottom template (3) are both provided with a plurality of uniformly distributed fishbone-shaped grooves (4), and the surfaces of the plurality of fishbone-shaped grooves (4) are all fixedly connected with herringbone-shaped fishbones (5).
2. The microfluidic chip for synthesizing mRNA lipid nanoparticles according to claim 1, wherein: The peak side angle of the herringbone-shaped fishbone (5) is 45°, the height of the herringbone-shaped fishbone (5) is 30μm, and the spacing between two herringbone-shaped fishbones (5) is 50μm.
3. The microfluidic chip for synthesizing mRNA lipid nanoparticles according to claim 1, wherein: The total height of the chip body (1) is 80μm.
4. The microfluidic chip for synthesizing mRNA lipid nanoparticles according to claim 1, wherein: The inside of the top template (2) is provided with two sample inlets (6), and the inside of the top template (2) is provided with a product outlet (7).
5. The microfluidic chip for synthesizing mRNA lipid nanoparticles according to claim 4, wherein: The length of the sample inlet (6) is 20mm, the included angle between the two sample inlets (6) is 120°, and the length of the product outlet (7) is 5mm.