A mixing unit, a microfluidic chip and a mixing system

Abstract

The application discloses a kind of 3D microfluidic chip for producing nanobiomedicine and its preparation product method and application, the mixing channel of 3D mixing unit includes first channel and second channel, the first channel and the second channel of mixing channel are located in the same plane, any adjacent two mixing channels are located in different planes.Because in 3D mixing unit, any adjacent two mixing channels are located in different planes, i.e. multiple mixing channels form 3D layout, so that when fluid in mixing channel enters the next mixing channel, it will enter another spatial plane, increasing the probability of turbulent flow and fluid components colliding with each other, can improve fluid mixing efficiency and improve mixing effect, to solve various applications such as lipid nanoparticle water phase and organic phase mixing efficiency and poor mixing effect problem, to accurately control nanoparticle production, improve drug embedding efficiency, improve drug delivery and treatment effect.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, specifically to a hybrid unit, a microfluidic chip, and a hybrid system. Background Technology

[0002] Nanoparticles are widely used for the delivery of biomolecules to prepare vaccines and biopharmaceuticals for disease prevention and treatment. Biomacromolecules such as proteins and nucleic acid drugs have more clearly defined disease mechanisms and therapeutic effects, and have seen faster development in recent years. However, protein and nucleic acid drugs, especially nucleic acid drugs, have poor stability, requiring effective delivery technologies to protect the drug and enhance efficacy. Lipid nanoparticles have been developed in recent years for the delivery of nucleic acid drugs, including mRNA, siRNA, and ASO.

[0003] Lipid nanoparticles require mixing aqueous and organic phases to form stable, high-quality nanoparticles, making their production challenging. In recent years, various microfluidic mixers have been widely used in the production of lipid nanoparticles, but different designs exhibit significant differences in mixing efficiency, nanoparticle quality, production scale-up, and cost. Summary of the Invention

[0004] This invention provides a mixing unit, a microfluidic chip, and a mixing system to solve the problems of low mixing efficiency and poor mixing effect of lipid nanoparticles in aqueous and organic phases.

[0005] In one embodiment, a mixing unit is provided, comprising a plurality of mixing channels connected in sequence: The hybrid channel includes a first channel and a second channel, wherein the inlet of the first channel is connected to the inlet of the second channel to form a common inlet, and the outlet of the first channel is connected to the outlet of the second channel to form a common outlet; The common inlet of the mixing channel at the foremost end is used to inject at least the first raw material and the second raw material, and the common outlet of the mixing channel at the rear end is used to discharge the mixture formed by the mixing of the first raw material and the second raw material. The common inlet of the mixing channel in the middle is connected to the common outlet of the adjacent mixing channel, and the common outlet of the mixing channel in the middle is connected to the common inlet of the adjacent mixing channel. The first and second channels of the hybrid channel are located in the same plane, and any two adjacent hybrid channels are located in different planes.

[0006] In one embodiment, the included angle between the planes of two adjacent mixing channels is 15°-165°.

[0007] In one embodiment, the included angle between the planes of two adjacent mixing channels is 45°-135°.

[0008] In one embodiment, the included angle between the planes of two adjacent mixing channels is 90°.

[0009] In one embodiment, the plurality of the mixing channels are arranged spirally along a straight line.

[0010] In one embodiment, the first channel and / or the second channel are bent, and the angle between the inlet of the first channel and the inlet of the second channel is 15°-165°, and the angle between the outlet of the first channel and the outlet of the second channel is 15°-165°.

[0011] In one embodiment, the angle between the inlet of the first channel and the inlet of the second channel is 45-135°, and the angle between the outlet of the first channel and the outlet of the second channel is 45°-135°.

[0012] In one embodiment, the angle between the inlet of the first channel and the inlet of the second channel is 90°, and the angle between the outlet of the first channel and the outlet of the second channel is 90°.

[0013] In one embodiment, the inner diameters of the first channel and the second channel are 5.0 mm to 0.05 mm; Alternatively, the inner diameters of the first channel and the second channel are 2.0 mm to 0.1 mm; Alternatively, the inner diameter of the first channel and the second channel is 1.5mm-0.2mm.

[0014] In one embodiment, the inner diameters of the first channel and the second channel satisfy the following regression formula requirement with respect to the particle size of the mixture: y = -17.612x1 2 +72.706x1+47.987 Where x1 is the inner diameter of the first channel and the second channel, and y is the particle size of the mixture.

[0015] In one embodiment, a microfluidic chip is provided, comprising: a first inlet channel, a second inlet channel, an outlet channel, and the aforementioned mixing unit. The first inlet channel and the second inlet channel are connected to a common inlet of the mixing channel located at the foremost end, and the outlet channel is connected to a common outlet of the mixing channel located at the rearmost end.

[0016] In one embodiment, a third entrance channel is further included, wherein the exits of the first entrance channel and the second entrance channel are connected to the entrance of the third entrance channel, and the exit of the third entrance channel is connected to the common entrance of the hybrid channel located at the foremost end.

[0017] In one embodiment, the first inlet channel and / or the second inlet channel are distributed on different axes from the third inlet channel.

[0018] In one embodiment, the angle between the first inlet channel and / or the second inlet channel and the third inlet channel is 15°-165°.

[0019] In one embodiment, the angle between the first inlet channel and / or the second inlet channel and the third inlet channel is 90°.

[0020] In one embodiment, the system further includes a diversion channel and a merging channel, with a plurality of mixing units disposed between the diversion channel and the merging channel; the diversion channel includes one or two inlets and a plurality of outlets, the first inlet channel and the second inlet channel being connected to the inlet of the diversion channel, and the plurality of outlets of the diversion channel being connected to a plurality of mixing units in a one-to-one correspondence; the merging channel includes a plurality of inlets and an outlet, the plurality of inlets of the merging channel being connected to a plurality of mixing units in a one-to-one correspondence, and the outlet of the merging channel being connected to the outlet channel.

[0021] In one embodiment, a plurality of the mixing units are arranged in parallel at intervals.

[0022] In one embodiment, the microfluidic chip further includes a substrate, and the first inlet channel, the second inlet channel, the outlet channel, and the mixing unit are located within the substrate.

[0023] In one embodiment, the substrate is a flat plate structure, and the inlet of the first inlet channel, the inlet of the second inlet channel, and the outlet of the outlet channel are located on the same side of the substrate.

[0024] In one embodiment, a first fluid connector, a second fluid connector, and a third fluid connector are mounted on the same side of the substrate. The first fluid connector is connected to the inlet of the first inlet channel, the second fluid connector is connected to the inlet of the second inlet channel, and the third fluid connector is connected to the outlet of the outlet channel.

[0025] In one embodiment, the first fluid connector, the second fluid connector, and the third fluid connector are female Luer type fluid connectors.

[0026] In one embodiment, a hybrid system is provided, comprising: The aforementioned microfluidic chip; A first raw material supply device is used to store a first raw material. The first raw material supply device is connected to the first inlet channel and is used to inject the first raw material into the first inlet channel. A second raw material supply device, used for storing a second raw material, is connected to a second inlet channel and is used to inject the second raw material into the second inlet channel; and A collection device, connected to the outlet channel, is used to collect the mixture generated by mixing the first raw material and the second raw material.

[0027] In one embodiment, a driving device is further included, which is connected to the first raw material supply device and the second raw material supply device. The driving device is used to drive the first raw material to be injected into the first inlet channel and to drive the second raw material to be injected into the second inlet channel.

[0028] In one embodiment, the system further includes a cleaning device and a waste liquid device. The cleaning device is connected to the first inlet channel and the second inlet channel and is used to inject cleaning fluid. The waste liquid device is connected to the outlet channel and is used to collect cleaning waste liquid.

[0029] In one embodiment, a method for preparing a product using the above-described microfluidic chip or the above-described hybrid system is provided, comprising the following steps: The first raw material and the second raw material are respectively injected into the microfluidic chip; The first raw material and the second raw material are mixed in the mixing unit to form a mixture.

[0030] In one embodiment, the flow rates of the first and second raw materials within the mixing unit satisfy the following regression equation with respect to the particle size of the mixture: y=0.0718x2 2 -5.4452x2+185.47 Where x2 is the flow rate of the first raw material and the second raw material in the mixing unit, and y is the particle size of the mixture.

[0031] In one embodiment, an application is provided in the preparation of products using the microfluidic chip or the hybrid system described above; Alternatively, the product may be particles or a pharmaceutical composition carrying drug molecules or cells; Alternatively, the particles may be lipid nanoparticles for drug delivery.

[0032] According to the above embodiments, the mixing unit, microfluidic chip, and mixing system, since any two adjacent mixing channels in the mixing unit are located in different planes, that is, multiple mixing channels form a 3D layout, when the fluid in the mixing channel enters the next mixing channel, it will enter another spatial plane, which increases the probability of turbulence and collision between fluid components, thereby improving the fluid mixing efficiency and mixing effect, and thus solving the problems of low mixing efficiency and poor mixing effect of lipid nanoparticles in aqueous and organic phases. Attached Figure Description

[0033] Figure 1 This is a schematic diagram of the structure of the hybrid unit in one embodiment; Figure 2 This is a schematic diagram of the structure of a microfluidic chip in one embodiment; Figure 3 This is a schematic diagram of the structure of a microfluidic chip in one embodiment; Figure 4 This is a diagram showing the relationship between the chip structure, LNP particles, and PDI in one embodiment; Figure 5 This is a diagram showing the relationship between the chip inner diameter and LNP particles and PDI in one embodiment; Figure 6 This is a diagram showing the relationship between the chip's inner diameter and the LNP particles in one embodiment; Figure 7 This is a schematic diagram of the structure of a microfluidic chip in one embodiment; Figure 8 This is a structural block diagram of a hybrid system in one embodiment; Figure 9 This is a structural block diagram of a hybrid system in one embodiment; Figure 10 This is a structural block diagram of a hybrid system in one embodiment; Figure 11 This is a graph showing the relationship between the preparation flow rate and LNP particles and PDI in one embodiment; Figure 12 This is a graph showing the relationship between the flow rate and LNP particles in one embodiment; The accompanying diagrams are labeled as follows: 10-Mixing unit, 11-Mixing channel, 111-First channel, 112-Second channel, 20-First inlet channel, 30-Second inlet channel, 40-Outlet channel, 50-Third inlet channel, 61-First fluid connector, 62-Second fluid connector, 63-Third fluid connector, 70-Diverting channel, 80-Merging channel; 100-Microfluidic chip, 101-Substrate, 200-First raw material supply device, 300-Second raw material supply device, 400-Collection device, 500-Drive device, 600-Cleaning device, 700-Waste liquid device; P1 - the plane where the previous blending channel is located, P2 - the plane where the next blending channel is located. Detailed Implementation

[0034] The present invention will now be described in further detail with reference to specific embodiments and accompanying drawings. Similar elements in different embodiments are referred to by associated similar element reference numerals. In the following embodiments, many details are described to facilitate a better understanding of this application. However, those skilled in the art will readily recognize that some features may be omitted in different situations, or may be replaced by other elements, materials, or methods. In some cases, certain operations related to this application are not shown or described in the specification. This is to avoid obscuring the core parts of this application with excessive description. For those skilled in the art, detailed description of these related operations is not necessary; they can fully understand the related operations based on the description in the specification and general technical knowledge in the art.

[0035] Furthermore, the features, operations, or characteristics described in the specification can be combined in any suitable manner to form various embodiments. At the same time, the order of the steps or actions in the method description can be changed or adjusted in a manner obvious to those skilled in the art. Therefore, the various orders in the specification and drawings are only for the clear description of a particular embodiment and do not imply a necessary order, unless otherwise stated that a particular order must be followed.

[0036] The serial numbers assigned to components in this document, such as "first" and "second," are used only to distinguish the described objects and have no sequential or technical meaning. The terms "connection" and "linkage" used in this application, unless otherwise specified, include both direct and indirect connections (linkages). Example 1:

[0037] This embodiment provides a mixing unit for mixing multiple raw materials to obtain a mixture. The first raw material and the second raw material can be mixed to obtain a mixture, or a larger number of different raw materials can be injected into the mixing unit to obtain a mixed product, hereinafter referred to as a mixture. For example, mixing an aqueous phase and an organic phase can obtain lipid nanoparticles (LNPs). This mixing unit can also mix other substances to prepare vaccines or other biological drugs.

[0038] Please refer to Figure 1 The mixing unit 10 in this embodiment mainly includes a plurality of mixing channels 11 connected in sequence, and the plurality of mixing channels 11 are connected end to end in sequence to form a single chain structure.

[0039] The hybrid channel 11 includes a first channel 111 and a second channel 112. The first channel 111 and the second channel 112 are combined to form a structure in which the head and tail are connected and the middle part is separated. The first channel 111 and the second channel 112 are combined to form a square, circular or elliptical structure.

[0040] The first channel 111 and the second channel 112 have entrances and exits at their respective ends. The entrance of the first channel 111 is connected to the entrance of the second channel 112 to form a common entrance, and the exit of the first channel 111 is connected to the exit of the second channel 112 to form a common exit.

[0041] Multiple mixing channels 11 are connected end-to-end in sequence. The common inlet of the mixing channel 11 at the foremost end is used to inject the first and second raw materials, and the common outlet of the mixing channel at the rearmost end is used to discharge the mixture formed by the first and second raw materials. The common inlet of the mixing channel 11 between the foremost and rearmost ends is connected to the common outlet of the preceding mixing channel 11, and the common outlet of the mixing channel 11 between the foremost and rearmost ends is connected to the common inlet of the following mixing channel 11. The first and second raw materials will be mixed sequentially in the multiple mixing channels 11.

[0042] In this embodiment, the first channel 111 and the second channel 112 of each mixing channel 11 are located in the same plane, and the central axis of the first channel 111 and the central axis of the second channel 112 are located in the same plane. Two adjacent mixing channels 11 are located in different planes. For example, the first mixing channel 11 is located in plane P1, and the second mixing channel 11 is located in plane P2. There is an angle between plane P1 and plane P2. They are not coplanar and are not parallel.

[0043] The included angle between plane P1 and plane P2 is 15°-165°. For example, the included angle between plane P1 and plane P2 is 15°, 25°, 30°, 45°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 135°, 145°, 155° and 165°, etc., which ensures that plane P1 and plane P2 are at least ±15° apart. This ensures that when the first and second raw materials mixed in the previous mixing channel 11 enter the next mixing channel 11, they will undergo at least a 15° turning collision, thereby improving the mixing efficiency and uniformity of the first and second raw materials.

[0044] In a preferred embodiment, the included angle between plane P1 and plane P2 is 45°-135°, for example, the included angle between plane P1 and plane P2 is 45°, 60°, 70°, 80°, 90°, 100°, 110°, 120° and 135°, etc., that is, to ensure that plane P1 and plane P2 are at least ±45° apart, so that when the first raw material and the second raw material mixed in the previous mixing channel 11 enter the next mixing channel 11, they will undergo at least a 45° turning collision, which significantly improves the mixing efficiency and uniformity of the first raw material and the second raw material.

[0045] In a more preferred embodiment, the angle between plane P1 and plane P2 is 90°, so that when the first and second raw materials mixed in the previous mixing channel 11 enter the next mixing channel 11, they will undergo a 90° turn and collision, which significantly improves the mixing efficiency and uniformity of the first and second raw materials.

[0046] In this embodiment, multiple mixing channels 11 are spirally arranged along a straight line, which is beneficial for the layout of the mixing units. Furthermore, the included angles between the multiple mixing channels 11 are arranged in a sequentially increasing or decreasing manner. For example, the angles between the planes containing the multiple mixing channels 11 and the horizontal plane, from the front end to the back end, are 0°, 90°, 180°, 270°, 0°… arranged spirally. This arrangement ensures that the mixing units are distributed along a straight line, facilitating the side-by-side arrangement of multiple mixing units 10. The planes containing adjacent mixing channels 11 have the same included angle, allowing the mixing units to produce different numbers of mixing channels 11.

[0047] In other embodiments, the multiple mixing channels 11 may also be arranged spirally along a curve, and the planes where adjacent mixing channels 11 are located may have different included angles to meet the needs of different application scenarios.

[0048] In this embodiment, one or both of the first channel 111 and the second channel 112 of the mixing channel 11 can be bent, and the first channel 111 and the second channel 112 can be bent into a broken line or a curve. The bending of at least one of the first channel 111 and the second channel 112 allows for collision mixing when the first channel 111 and the second channel 112 meet, thereby improving the mixing effect.

[0049] The angle between the inlet of the first channel 111 and the inlet of the second channel 112 is 15°-165°, for example, the angle is 15°, 25°, 30°, 45°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 135°, 145°, 155° and 165°, etc., to ensure that the inlet of the first channel 111 and the inlet of the second channel 112 differ by at least ±15°, so that when the first raw material and the second raw material meet, they can achieve collision mixing, and will undergo at least a 15° turning collision, thereby improving the mixing efficiency and uniformity of the first raw material and the second raw material.

[0050] In a preferred embodiment, the angle between the inlet of the first channel 111 and the inlet of the second channel 112 is 45°-135°, for example, the angle is 45°, 60°, 70°, 80°, 90°, 100°, 110°, 120° and 135°, etc., to ensure that the inlet of the first channel 111 and the inlet of the second channel 112 differs by at least ±45°, so that when the first raw material and the second raw material meet, they can achieve collision mixing, and will collide after at least 45° of turning collision, which significantly improves the mixing efficiency and uniformity of the first raw material and the second raw material.

[0051] In a more preferred embodiment, the angle between the inlet of the first channel 111 and the inlet of the second channel 112 is 90°, ensuring that the inlet of the first channel 111 and the inlet of the second channel 112 are 90° apart, so that the first raw material and the second raw material can be mixed by collision when they meet. After the 90° turn and collision, the mixing efficiency and uniformity of the first raw material and the second raw material are significantly improved.

[0052] In this embodiment, the inner diameter of the first channel 111 and the second channel 112 is related to the particle size of the resulting mixture. The inner diameter of the first channel 111 and the second channel 112 can be designed according to the mixture to be produced. For example, the inner diameter of the first channel 111 and the second channel 112 is 1.5mm-0.2mm. The inner diameters of the first channel 111 and the second channel 112 can be the same or different.

[0053] In this embodiment, the mixing unit 10 has two adjacent mixing channels 11 located in different planes, forming a 3D layout. This allows the fluid in the mixing channel 11 to enter another spatial plane when it enters the next mixing channel 11, increasing the probability of fluid collisions and improving the mixing efficiency and effect. This solves the problems of low mixing efficiency and poor mixing effect between lipid nanoparticles in aqueous and organic phases. Example 2:

[0054] Please refer to Figure 2This embodiment provides a microfluidic chip 100, which includes a first inlet channel 20, a second inlet channel 30, an outlet channel 40, and the mixing unit 10 in the above embodiment 1.

[0055] The first inlet channel 20 and the second inlet channel 30 are located at the front end of the mixing unit 10. The outlets of the first inlet channel 20 and the second inlet channel 30 are connected to the common inlet of the foremost mixing channel 11. The first inlet channel 20 is used to inject a first raw material into the mixing channel 11, and the second inlet channel 30 is used to inject a second raw material into the mixing channel 11. The outlet channel 40 is located at the rear end of the mixing channel 11. The inlet of the outlet channel 40 is connected to the common outlet of the rearmost mixing channel 11, and the outlet channel 40 is used to discharge the mixture formed by mixing.

[0056] The microfluidic chip 100 also includes a substrate 101, which can be a flat plate structure. The mixing unit 10, the first inlet channel 20, the second inlet channel 30 and the outlet channel 40 are located inside the substrate 101. The inlet of the first inlet channel 20, the inlet of the second inlet channel 30 and the outlet of the outlet channel 40 can be exposed on the same side of the substrate 101 to realize the injection of the first raw material and the second raw material, as well as the discharge of the mixture.

[0057] The substrate 101 may include an upper plate and a lower plate, which are combined to form a mixing unit 10, a first inlet channel 20, a second inlet channel 30, and an outlet channel 40. The substrate 101 may also be an integral structure, with the mixing unit 10, the first inlet channel 20, the second inlet channel 30, and the outlet channel 40 fabricated on the substrate 101 by injection molding or 3D printing.

[0058] Please refer to Figure 2 In this embodiment, the microfluidic chip 100 may further include a third inlet channel 50, which is a transition channel. The first inlet channel 20 and the second inlet channel 30 are connected to the mixing unit 10 through the third inlet channel 50. The inlet of the third inlet channel 50 is connected to the outlet of the first inlet channel 20 and the outlet of the second inlet channel 30, and the outlet of the third inlet channel 50 is connected to the common inlet of the mixing channel 11 located at the foremost end of the mixing unit 10. The first raw material and the second raw material can be pre-mixed in the third inlet channel 50. The setting of the third inlet channel 50 can improve the uniformity of the mixing of the first raw material and the second raw material.

[0059] The third inlet channel 50 is distributed along different axes from the first inlet channel 20 and the second inlet channel 30, so that the connection between the first inlet channel 20, the second inlet channel 30 and the third inlet channel 50 forms a turning point, so that the first raw material and the second raw material can collide when they meet, thereby achieving a certain mixing effect.

[0060] The angle between the first entrance channel 20 and the third entrance channel 50 can be 15°-165°, and the angle between the second entrance channel 30 and the third entrance channel 50 can be 15°-165°.

[0061] For example, the angle between the first inlet channel 20 and the third inlet channel 50 can be 90°, and the angle between the second inlet channel 30 and the third inlet channel 50 can be 90°. The third inlet channel 50 forms a T-shaped distribution with the first inlet channel 20 and the second inlet channel 30, which can significantly improve the mixing effect of the first raw material and the second raw material.

[0062] In other embodiments, the third inlet channel 50 is at an angle to one of the first inlet channel 20 and the second inlet channel 30, and is parallel or collinear with the other, which can also achieve a certain mixing effect between the first raw material and the second raw material.

[0063] In other embodiments, the microfluidic chip 100 may also exclude the third inlet channel 50, and the first inlet channel 20 and the second inlet channel 30 may be directly connected to the mixing unit 10. Through the mixing of the mixing unit 10, multiple raw materials can also be mixed to obtain a mixture.

[0064] Please refer to Figure 3 In this embodiment, a first fluid connector 61, a second fluid connector 62, and a third fluid connector 63 are mounted on the same side of the substrate 101 of the microfluidic chip 100. The first fluid connector 61, the second fluid connector 62, and the third fluid connector 63 can be female Luer type fluid connectors or other connectors with good sealing connection effect.

[0065] The first fluid connector 61, the second fluid connector 62, and the third fluid connector 63 are respectively vertically installed on the same side of the base 101. The first fluid connector 61 is connected to the inlet of the first inlet channel 20, the second fluid connector 62 is connected to the inlet of the second inlet channel 30, and the third fluid connector 63 is connected to the outlet of the third inlet channel 50. The first fluid connector 61, the second fluid connector 62, and the third fluid connector 63 serve as adapters. The first fluid connector 61 is used to connect to a device containing a first raw material, the second fluid connector 62 is used to connect to a device containing a second raw material, and the third fluid connector 63 is used to connect to a device for collecting the mixture.

[0066] In other embodiments, the first fluid connector 61, the second fluid connector 62, and the third fluid connector 63 may also be disposed on different sides of the base 101. For example, the first fluid connector 61 and the second fluid connector 62 may be disposed on the same side of the base 101, and the third fluid connector 63 may be disposed on the other side of the base 101. Correspondingly, the inlet of the first inlet channel 20 and the inlet of the second inlet channel 30 are located on the same side of the base 101, and the outlet of the outlet channel 40 is located on the other side of the base 101.

[0067] In this embodiment, the microfluidic chip includes a 3D hybrid unit 10, making the microfluidic chip a 3D microfluidic chip.

[0068] The differences between the 3D microfluidic chip and the traditional 2D ring microfluidic mixer in LNP preparation in this embodiment were investigated. Four chip sizes were used: 2D-0.2 mm-9 rings, 2D-0.5 mm-9 rings, 3D-0.2 mm-8 rings, and 3D-0.2 mm-16 rings. The effects of these four chips on empty LNP preparation were compared at flow rates of 4 ml / min, 8 ml / min, 12 ml / min, and 16 ml / min, respectively. Furthermore, the effects on mRNA-LNP preparation were compared. The number of rings refers to the number of mixing channels 11; for example, 9 rings means that the mixing unit 10 includes 9 mixing channels 11.

[0069] Please refer to Figure 4 ,from Figure 4 The comparison shows that for the same chip, the LNP particle size gradually decreases with increasing flow rate; the 0.2mm particle size of the 2D chip is smaller than that of the 0.5mm chip. Similarly, for the same 0.2mm particle size, the 3D chip has a smaller particle size than the 2D chip. Also, for the same 0.2mm 3D chip, the 16-ring particle size is slightly smaller than the 8-ring particle size.

[0070] To further compare the effects of 3D and 2D, 3D-0.2mm and 2D-0.2mm were used to prepare mRNA-LNP under the same mRNA, lipid, and process conditions. The results are shown in Table 1. Before and after purification, the 3D particles were smaller than the 2D particles, and the encapsulation efficiency of the 3D particles was significantly higher than that of the 2D particles.

[0071] Table 1. Comparison of mRNA-LNP preparation results between 3D-0.2mm and 2D-0.2mm.

[0072] Referring to Table 1 above, it is clear that compared with the traditional 2D toroidal microfluidic mixer, the 3D microfluidic chip can produce a mixture with a smaller particle size and has a better mixing effect.

[0073] The microfluidic chip 100 of this embodiment has an inner diameter that can be set to 5.0mm-0.05mm, meaning the inner diameters of the first channel 111 and the second channel 112 are 5.0mm-0.05mm. A preferred embodiment is that the inner diameters of the first channel 111 and the second channel 112 are 2.0mm-0.1mm; and a more preferred embodiment is that the inner diameters of the first channel 111 and the second channel 112 are 1.5mm-0.2mm. For example, the inner diameters of the first channel 111 and the second channel 112 can be 0.2mm, 0.5mm, 0.75mm, 1.0mm, 1.25mm, and 1.5mm to more precisely produce LNPs of different sizes to match the delivery requirements of different tissues and organs.

[0074] Targeted delivery to tissues and organs is a key objective of drug delivery. Passive tissue localization primarily depends on the size and charge of low-density polymeric nipple (LNP) particles. The LNP particle size is influenced by the microfluidic chip, lipid composition, and manufacturing process conditions. Please refer to [reference needed]. Figure 5 This study demonstrates the impact of chip inner diameter on LNP particle size and PDI. Under the same resin and process conditions, as the chip inner diameter decreases, the produced LNP particle size also decreases. From an inner diameter of 1.5 mm to 0.2 mm, the LNP particle size decreases from ~120 nm to ~60 nm, while the PDI is controlled below 0.1, reflecting a high degree of particle uniformity. Please refer to [reference needed]. Figure 6 The trend lines of LNP particle size y and chip inner diameter x satisfy the following regression equation: y = -17.612x1 2 + 72.706x1 + 47.987 Where x1 is the inner diameter of the microfluidic chip 100, that is, x1 is the inner diameter of the first channel 111 and the second channel 112, and y is the particle size of the mixture (LNP particle size).

[0075] The regression equation above achieves a very high regression coefficient R0. 2 = 0.9965. Therefore, LNPs of different sizes can be precisely manufactured based on the chip's inner diameter and process control, as shown in Table 2.

[0076] Table 2. Comparison of theoretical and experimental values ​​of LNP particle size

[0077] The microfluidic chip 100 in this embodiment has a 3D structure. The 3D microfluidic chip 100 is equipped with a 3D mixing unit 10. Any two adjacent mixing channels 11 are located in different planes, that is, multiple mixing channels 11 form a 3D layout. When the fluid in the mixing channel 11 enters the next mixing channel 11, it will enter another spatial plane, which increases the probability of fluid collision and can improve the fluid mixing efficiency and mixing effect. Example 3:

[0078] This embodiment provides a microfluidic chip 100. The difference between this microfluidic chip 100 and the above embodiment 2 is that the microfluidic chip 100 in this embodiment includes multiple mixing units 10, which can increase the throughput of the microfluidic chip 100 and improve production efficiency.

[0079] Please refer to Figure 7 In this embodiment, the microfluidic chip 100 further includes a diversion channel 70 and a merging channel 80. A plurality of mixing units 10 are disposed between the diversion channel 70 and the merging channel 80. The front end of the diversion channel 70 is connected to a first inlet channel 20 and a second inlet channel 30, and the rear end of the merging channel 80 is connected to an outlet channel 40.

[0080] The diversion channel 70 includes one or two inlets and multiple outlets. The first inlet channel 20 and the second inlet channel 30 are connected to one inlet of the diversion channel 70, or the first inlet channel 20 and the second inlet channel 30 are each connected to one of the two inlets of the diversion channel 70. The number of outlets of the diversion channel 70 is the same as the number of mixing units 10, and the multiple outlets of the diversion channel 70 are connected to multiple mixing units 10 in a one-to-one correspondence. The merging channel 80 includes multiple inlets and one outlet. The number of inlets of the merging channel 80 is the same as the number of mixing units 10, and the multiple inlets of the merging channel 80 are connected to multiple mixing units 10 in a one-to-one correspondence. The outlet of the merging channel 80 is connected to the outlet channel 40.

[0081] For example, the microfluidic chip 100 includes 5 mixing units 10, and the diversion channel 70 has 5 outlets and the confluence channel 80 has 5 outlets connected to them, so that the 5 mixing units 10 can mix and produce mixtures at the same time.

[0082] Multiple mixing units 10 within the microfluidic chip 100 can be arranged in parallel at intervals, with the central axes of the mixing units 10 distributed in the same plane. This makes the layout of the multiple mixing units 10 more compact, enabling the microfluidic chip 100 to achieve high throughput and miniaturization. Of course, the multiple mixing units 10 can also be arranged in multiple parallel planes.

[0083] The 3D microfluidic chip 100 in this embodiment uses a chip with the same mechanism to solve the problem of production scale-up, ensuring the repeatability of the process and quality before and after scale-up, and avoiding adverse effects on the quality of the scaled-up product. For most applications, the ideal LNP particle size range is about 80-120nm, with a typical initial size of about 80nm, gradually increasing during the production process and storage and transportation. As shown in Table 3, the ideal preparation flow rate should be 40ml / min, which is equivalent to 2.4L / hour. Based on 4 hours / batch, 9.6L / batch can be produced. Based on 0.1ml / dose vaccine, this is equivalent to 96,000 doses / batch, which is sufficient to meet clinical needs. For large-scale commercial production, we designed a unique parallel chip. Figure 7 Taking the 5-cell hybrid unit 10 as an example, the consistent structure and function of each channel ensures product quality stability during production scale-up. Using the 5-cell chip, at a batch rate of 10 hours / batch, 2.4 × 5 × 10 = 96 L can be produced, equivalent to 1.2 million doses / batch at 0.1 ml / dose. In an emergency, at one batch per day, 330 batches per year could produce 396 million doses. The 5-cell chip can be further augmented with parallel channels to increase production capacity. Another scale-up approach is to simply repeat the production of the 5-cell chip; for example, three 5-cell chips can produce 1.188 billion doses, which can meet the needs of large-scale vaccine administration in emergency situations.

[0084] Table 3. Comparison of scale-up routes for laboratory preparation, clinical production, and commercial production of microfluidic chip 100

[0085] As shown in Table 3 above, the multi-channel sampling microfluidic chip 100 can greatly increase production to meet market demand. Example 4:

[0086] Please refer to Figure 8 This embodiment provides a mixing system, which includes the microfluidic chip 100 in the above embodiment three or four, and also includes a first raw material supply device 200, a second raw material supply device 300 and a collection device 400. When the microfluidic chip 100 needs more raw materials to be mixed, the mixing system can also be provided with more raw material supply devices.

[0087] The first raw material supply device 200 is used to store the first raw material. The first raw material supply device 200 is connected to the first fluid connector 61 through a pipe. The first raw material supply device 200 is used to inject the first raw material into the microfluidic chip 100.

[0088] The second raw material supply device 300 is used to store the second raw material. The second raw material supply device 300 is connected to the second fluid connector 62 through a pipe. The second raw material supply device 300 is used to inject the second raw material into the microfluidic chip 100.

[0089] The collection device 400 is connected to the third fluid connector 63 via a pipe, and the collection device 400 is used to collect the mixture prepared by mixing the microfluidic chip 100.

[0090] Please refer to Figure 9 In this embodiment, the mixing system may further include a driving device 500. The driving device 500 may include two devices: one connected to the first raw material supply device 200 for driving the injection of the first raw material into the microfluidic chip 100, and the other connected to the second raw material supply device 300 for driving the injection of the second raw material into the microfluidic chip 100. The driving device 500 enables automated injection of the first and second raw materials, control of the injection ratio of the first and second raw materials, and control of the injection flow rate of the first and second raw materials.

[0091] In other embodiments, the first raw material supply device 200 and the second raw material supply device 300 are equipped with driving components, which can also realize the automatic injection of the first raw material and the second raw material.

[0092] Please refer to Figure 10 In other embodiments, the mixing system may further include a cleaning device 600 and a waste liquid device 700. The cleaning device 600 is used to store cleaning fluid and is connected to a first fluid connector 61 and a second fluid connector 62 of the microfluidic chip 100 via pipes. The cleaning device 600 is used to inject cleaning fluid into the microfluidic chip 100 to clean the flow channels within the microfluidic chip 100. The waste liquid device 700 is connected to a third fluid connector 63 via pipes and is used to collect cleaning waste liquid.

[0093] The pipelines connected by the first fluid connector 61, the second fluid connector 62, and the third fluid connector 63 are equipped with multi-port valves, which can be used to switch the injection of raw materials or cleaning fluid, as well as to switch the discharge of mixed liquid or waste liquid to different devices.

[0094] In this embodiment, the mixing system includes the microfluidic chip 100 in the above embodiment. Since any two adjacent mixing channels 11 in the mixing unit 10 are located in different planes, that is, multiple mixing channels 11 form a 3D layout, when the fluid in the mixing channel 11 enters the next mixing channel 11, it will enter another spatial plane, which increases the probability of fluid collision and can improve the fluid mixing efficiency and mixing effect. Example 5:

[0095] This embodiment provides a method for preparing a product by mixing. This method is implemented using the microfluidic chip 100 or the mixing system described in the above embodiment.

[0096] This method mainly includes the following two steps: The first and second raw materials are respectively injected into the microfluidic chip 100; The first raw material and the second raw material are mixed in the mixing unit 10 to form a mixture.

[0097] The two steps described above can be automated by a controller to control the injection ratio of the first and second raw materials, as well as the injection speed of the first and second raw materials, thereby controlling the flow rate of the first and second raw materials being mixed.

[0098] The particle size of the product can be controlled by controlling the flow rate within the microfluidic chip 100.

[0099] Please refer to Figure 11 and Figure 12 With a fixed chip inner diameter, the process flow rate affects the LNP particle size (product); increasing the flow rate can significantly reduce the LNP particle size. With the same chip inner diameter and flow rate, increasing the number of rings (the number of mixing channels) can slightly reduce the LNP particle size.

[0100] Raw material parameters such as the N / P ratio (lipoRNA ratio) and PEG-lipoprotein ratio can affect LNP particle size, but changes in these raw material parameters can significantly alter the efficacy and safety of the product, thus requiring strict control within a small range of variation. With raw material parameters remaining constant, chip inner diameter and flow rate become the key control factors for LNP particle size.

[0101] Linear regression was performed on the flow velocity and LNP particle size data of the 3D-0.5MM-12 ring, yielding Table 4 and the following regression equation: y=0.0718x2 2 -5.4452x2+185.47 Where x2 is the flow rate of the first and second raw materials in the mixing unit 10, y is the particle size of the mixture (LNP particle size), and the regression coefficient R of the above equation is... 2 = 0.9955.

[0102] Table 4. Calculated values ​​of LNP particle size for different flow rates

[0103] It is known that the faster the flow rate, the smaller the LNP particle size. The flow rate can be controlled to produce LNPs of different particle sizes to meet different production needs. Example 6:

[0104] This embodiment provides an application of the microfluidic chip 100 or the product prepared by the hybrid system in the above embodiments.

[0105] Microfluidic chip 100 can be used to produce particles or drug compositions carrying drug molecules or cells, such as lipid nanoparticles. Microfluidic chip 100 can also be used to produce other forms of nano or microparticles for application in various fields.

[0106] As shown in Table 5, the microfluidic chip 100 can be used to produce nano or micron-sized particles based on lipids or polymers for the delivery of small molecules, biomacromolecules, and cells for various medical, therapeutic, diagnostic, and detection applications.

[0107] Table 5. Application areas of 3D microfluidic chips

[0108] The above examples illustrate the present invention only to aid in understanding it and are not intended to limit the scope of the invention. Those skilled in the art can make various simple deductions, modifications, or substitutions based on the principles of this invention.

Claims

1. A hybrid unit, characterized in that, Includes multiple sequentially connected mixing channels: The hybrid channel includes a first channel and a second channel, wherein the inlet of the first channel is connected to the inlet of the second channel to form a common inlet, and the outlet of the first channel is connected to the outlet of the second channel to form a common outlet; The common inlet of the mixing channel at the foremost end is used to inject at least the first raw material and the second raw material, and the common outlet of the mixing channel at the rear end is used to discharge the mixture formed by the mixing of the first raw material and the second raw material. The common inlet of the mixing channel in the middle is connected to the common outlet of the adjacent mixing channel, and the common outlet of the mixing channel in the middle is connected to the common inlet of the adjacent mixing channel. The first and second channels of the hybrid channel are located in the same plane, any two adjacent hybrid channels are located in different planes, and there is an angle between the planes of the two adjacent hybrid channels.

2. The hybrid unit as described in claim 1, characterized in that, The included angle between the planes of two adjacent mixing channels is 15°-165°.

3. The hybrid unit as described in claim 2, characterized in that, The included angle between the planes of two adjacent mixing channels is 45°-135°.

4. The hybrid unit as described in claim 3, characterized in that, The included angle between the planes of two adjacent mixing channels is 90°.

5. The hybrid unit as claimed in claim 1, characterized in that, The multiple mixing channels are arranged spirally along a straight line.

6. The hybrid unit as claimed in claim 1, characterized in that, The first channel and / or the second channel are bent, and the angle between the inlet of the first channel and the inlet of the second channel is 15°-165°, and the angle between the outlet of the first channel and the outlet of the second channel is 15°-165°.

7. The hybrid unit as described in claim 6, characterized in that, The angle between the entrance of the first channel and the entrance of the second channel is 45-135°, and the angle between the exit of the first channel and the exit of the second channel is 45°-135°.

8. The hybrid unit as claimed in claim 7, characterized in that, The angle between the entrance of the first channel and the entrance of the second channel is 90°, and the angle between the exit of the first channel and the exit of the second channel is 90°.

9. The hybrid unit as claimed in claim 1, characterized in that, The inner diameters of the first channel and the second channel are 5.0mm-0.05mm; Alternatively, the inner diameters of the first channel and the second channel are 2.0 mm to 0.1 mm; Alternatively, the inner diameter of the first channel and the second channel is 1.5mm-0.2mm.

10. The hybrid unit as claimed in claim 9, characterized in that, The inner diameters of the first and second channels and the particle size of the mixture satisfy the following regression formula requirement: y=-17.612x1 2 +72.706x1+47.987 Where x1 is the inner diameter of the first channel and the second channel, and y is the particle size of the mixture.

11. A microfluidic chip, characterized in that, include: The system comprises a first inlet channel, a second inlet channel, an outlet channel, and a mixing unit as described in any one of claims 1 to 10, wherein the first inlet channel and the second inlet channel are connected to a common inlet of the mixing channel located at the foremost end, and the outlet channel is connected to a common outlet of the mixing channel located at the rearmost end.

12. The microfluidic chip as described in claim 11, characterized in that, It also includes a third entrance channel, the exit of the first entrance channel and the exit of the second entrance channel are connected to the entrance of the third entrance channel, and the exit of the third entrance channel is connected to the common entrance of the hybrid channel located at the foremost end.

13. The microfluidic chip as described in claim 12, characterized in that, The first inlet channel and / or the second inlet channel are distributed on different axes from the third inlet channel.

14. The microfluidic chip as described in claim 13, characterized in that, The angle between the first inlet channel and / or the second inlet channel and the third inlet channel is 15°-165°.

15. The microfluidic chip as described in claim 14, characterized in that, The angle between the first inlet channel and / or the second inlet channel and the third inlet channel is 90°.

16. The microfluidic chip as described in claim 11, characterized in that, It also includes a diversion channel and a merging channel, with multiple mixing units provided between the diversion channel and the merging channel; the diversion channel includes one or two inlets and multiple outlets, the first inlet channel and the second inlet channel are connected to the inlet of the diversion channel, and the multiple outlets of the diversion channel are connected to the multiple mixing units in a one-to-one correspondence; the merging channel includes multiple inlets and one outlet, the multiple inlets of the merging channel are connected to the multiple mixing units in a one-to-one correspondence, and the outlet of the merging channel is connected to the outlet channel.

17. The microfluidic chip as described in claim 16, characterized in that, The multiple mixing units are arranged in parallel at intervals.

18. The microfluidic chip according to any one of claims 11 to 17, characterized in that, The microfluidic chip also includes a substrate, and the first inlet channel, the second inlet channel, the outlet channel, and the mixing unit are located within the substrate.

19. The microfluidic chip as described in claim 18, characterized in that, The substrate is a flat plate structure, and the inlet of the first inlet channel, the inlet of the second inlet channel, and the outlet of the outlet channel are located on the same side of the substrate.

20. The microfluidic chip as described in claim 19, characterized in that, A first fluid connector, a second fluid connector, and a third fluid connector are installed on the same side of the substrate. The first fluid connector is connected to the inlet of the first inlet channel, the second fluid connector is connected to the inlet of the second inlet channel, and the third fluid connector is connected to the outlet of the outlet channel.

21. The microfluidic chip as described in claim 20, characterized in that, The first fluid connector, the second fluid connector, and the third fluid connector are female Luer type fluid connectors.

22. A hybrid system, characterized in that, include: The microfluidic chip as described in any one of claims 11 to 21; A first raw material supply device is used to store a first raw material. The first raw material supply device is connected to the first inlet channel and is used to inject the first raw material into the first inlet channel. The second raw material supply device is used to store the second raw material. The second raw material supply device is connected to the second inlet channel and is used to inject the second raw material into the second inlet channel. as well as A collection device, connected to the outlet channel, is used to collect the mixture generated by mixing the first raw material and the second raw material.

23. The hybrid system as claimed in claim 22, characterized in that, It also includes a driving device, which is connected to the first raw material supply device and the second raw material supply device. The driving device is used to drive the first raw material to be injected into the first inlet channel and to drive the second raw material to be injected into the second inlet channel.

24. The hybrid system as claimed in claim 22, characterized in that, It also includes a cleaning device and a waste liquid device. The cleaning device is connected to the first inlet channel and the second inlet channel and is used to inject cleaning fluid. The waste liquid device is connected to the outlet channel and is used to collect cleaning waste liquid.

25. A method for preparing a product using a microfluidic chip as described in any one of claims 11 to 21 or a hybrid system as described in any one of claims 22 to 24, characterized in that, Includes the following steps: The first raw material and the second raw material are respectively injected into the microfluidic chip; The first raw material and the second raw material are mixed in the mixing unit to form a mixture.

26. The method as described in claim 25, characterized in that, The flow rates of the first and second raw materials within the mixing unit and the particle size of the mixture satisfy the following regression equation: y=0.0718x2 2 -5.4452x2+185.47 Where x2 is the flow rate of the first raw material and the second raw material in the mixing unit, and y is the particle size of the mixture.

27. An application of a product prepared using a microfluidic chip as described in any one of claims 11 to 21 or a hybrid system as described in any one of claims 22 to 24; Alternatively, the product may be particles or a pharmaceutical composition carrying drug molecules or cells; Alternatively, the particles may be lipid nanoparticles for drug delivery.

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