Method, device, kit, and system for manufacturing particles by centrifugation

Abstract

The present invention relates to a method for manufacturing particles by centrifugation, using one or more devices, each comprising at least two ports for receiving solutions and one channel comprising at least one mixing section; wherein the solutions enter the mixing section when a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device(s), resulting in particles. The present invention also relates to the device, a kit comprising the device, and a system comprising the device and a centrifuge.

Description

[0001] METHOD, DEVICE, KIT, AND SYSTEM FOR MANUFACTURING PARTICLES BY CENTRIFUGATION

[0002] BACKGROUND OF THE IN VENTION

[0003] 1. Field of the Invention

[0004] The present invention relates to a method for manufacturing particles by centrifugation. The present invention also relates to a device that is configured to manufacture particles by centrifugation; and a kit and a system comprising the device.

[0005] 2. Background

[0006] Particles at the nano- and micro-scales are widely utilized across medicine, research, and the cosmetics industry. In drug delivery, many therapeutic materials can either exhibit toxicity or fail to reach target cells. Encapsulation of these therapeutic materials within particles comprising polymers, lipids, or other carrier materials can address these challenges. Such particle-based systems improve the therapeutic index by reducing off-target toxicity, enhancing delivery to diseased tissues, and enabling targeted delivery.

[0007] Among various methods for particle manufacturing, external pressure- driven microfluidic systems have demonstrated the ability to rapidly and controllably mix fluids to form nanoparticles. However, these conventional microfluidic systems I) are not widely accessible as they require specialized equipment such as pumps to control fluid flow, 2) are not high-throughput (preparation of multiple, distinc t particle formulations in short duration), and 3 ) are not suitable for small-volume (microliter to milliliter volumes) particle manufacturing that is particularly important to accelerate drug research and development. Although microfluidics mixing forms particles with significantly more desirable characteristics (e.g. smaller size, lower polydispersity) over particles formed by other conventional methods (e.g. manual mixing, vortex mixing, ethanol precipitation, ultrasonication, membrane extrusion), particle manufacturing in small volumes (microliter to milliliter volumes) for research, development, and point-of-use applications often rely on poorly controlled manual mixing techniques that yield particles with high variability and low reproducibility.

[0008] Challenges remain for manufacturing of high-quality particles at high throughput in small volumes with current microfluidics systems primarily due to the difficulty of uniformly controlling multiple small-volume fluid streams across parallelized devices. Existing systems require specialized equipment, such as syringe pumps, gear pumps, or peristaltic pumps and alike, to impart external pressure that propels fluids through microfluidic devices. These systems lead to long manufacturing duration, low throughput, and may not be readily available in certain locations, such as in resource-limited settings and at point-of-use. In addition, small-volume manufacturing is particularly affected by fluidic “wasted volume,” which includes hold-up volume within device channels and connecting lines. Such disadvantages present a substantial barrier to accessible, high-throughput, small-volume particle manufacturing.

[0009] The present invention addresses these challenges and provides additional related advantages by leveraging centrifugal force to flow fluids in micro fluidic device, so that particles can be robustly manufactured at high- throughput in small volumes using a conventional, widely accessible centrifuge without the need for specialized equipment.

[0010] SUMMARY OF THE INVENTION

[0011] The present invention provides a method for manufacturing particles by centrifugation, specifically a centrifugal microfluidic process for particle manufacturing that enables high-throughput, small-volume production of nanoparticles with improved accessibility and consistency. Specifically, the invention utilizes centrifugal force at or above 50xg to flow one or more fluid streams through a microfluidic device to facilitate the manufacturing of high- quality particles with minimal wasted volume.

[0012] Unlike traditional microfluidic systems that require pumps to apply external pressure to propel fluid flow, this invention relies solely on centrifugal force to induce fluid flow, enabling accessible, high-throughput, and robust execution for consistent small- volume nanoparticle manufacturing. A key benefit of the invention is high accessibility — requiring no specialized equipment, calibration, or maintenance. Additionally, the invention enables high-throughput small-volume particle manufacturing as multiple distinct particle formulations can be manufactured in parallel by centrifuging multiple devices simultaneously, regardless of individual capacities. This invention affords a robust process for particle manufacturing as uniform application of centrifugal force to all devices reduces variability across the resulting particles.

[0013] The microfluidic device described in this invention includes inlet ports that hold liquid solutions. Upon centrifugation, hydrostatic pressure generated by centrifugal force induces fluid flow through the device. The microfluidic device features channels oriented both in-plane and out-of-plane relative to the plane perpendicular to the axis of rotation, configured so that the principal direction of fluid flow is not directed counter to the direction of centrifugal force, enabling enhanced fluid interaction and improved mixing. Effective mixing is achieved through precise control of flow direction within channels with respect to the centrifugal force direction and flow resistance. One application of this invention is the manufacturing (e.g. assembly, modification) of nanoparticles that encapsulates therapeutics materials for drug delivery.

[0014] One objective of the present invention is to provide a method for manufacturing particles, comprising the following steps:

[0015] ( 1) providing a device for manufacturing particles, and the device comprising: a first port for receiving a first solution; a first channel comprising a first inlet in fluid communication with the first port; and a first outlet; a second port for receiving a second solution; a second channel comprising a second inlet in fluid communication with the second port: and a second outlet: a third channel comprising a third inlet in fluid communication with the first outlet and the second outlet; a third outlet; and a terminal mixing section between the third inlet and the third outlet and configured to mix fluid flow in the third channel to manufacture particles; and a first intersection of the first outlet, the second outlet and the third inlet;

[0016] (2) introducing a first solution into the first port, and a second solution into the second port, respectively;

[0017] (3) applying a relative centrifugal force (RCF) of 50xg to 32000xg to the device to initiate fluid flow in the device and manufacture particles.

[0018] Another objective of the present invention is to provide a device for manufacturing particles, and the device comprises: a first port for receiving a first solution; a first channel comprising a first inlet in fluid communication with the first port; and a first outlet; a second port for receiving a second solution; a second channel comprising a second inlet in fluid communication with the second port; and a second outlet; a third channel comprising a third inlet in fluid communication with the first outlet and the second outlet; a third outlet; and a terminal mixing section between the third inlet and the third outlet and configured to mix fluid flow in the third channel to manufacture particles; and a first intersection of the first outlet, the second outlet and the third inlet; wherein the first solution in the first port flows through the first channel and the second solution in the second port flows through the second channel when a relative centrifugal force (RCF) of 50* g to 32000xg is applied to the device.

[0019] Another objective of the present invention is to provide a kit for manufacturing particles, comprising the aforementioned device.

[0020] Still another objective of the present invention is to provide a system for manufacturing particles, comprising the aforementioned device and a centrifuge.

[0021] In some embodiments, the method comprises fluid flow and mixing on a rotating frame.

[0022] In some embodiments, the fluid flow and mixing are achieved only under a centrifugal force without the use of an external pressure source.

[0023] In some embodiments, the device further comprises: at least one addi tional port, and each of the at least one additional port for receiving an additional solution; at least one additional channel, and each of the at least one additional channel comprising: an additional inlet in fluid communication with a corresponding one of the at least one additional port; an additional outlet in fluid communication with the third channel; and wherein the additional solution in each of the at least one additional port flows through the at least one additional channel when the RCF of 50xg to 32000xg is applied to the device.

[0024] In some embodiments, the additional outlet of one or more of the at least one additional channel is connected to the first intersection.

[0025] In some embodiments, the third channel further includes at least one middle mixing section between the third inlet and the terminal mixing section; and the additional outlet of each of the at least one additional channel is connected to an intersection defined on the third channel and disposed between the terminal mixing section and one of the at least one middle mixing sec tion or between two of the middl e mixing section that are disposed next to each other when the at least one middle mixing section includes two or more middle mixing sections.

[0026] In some embodiments, the third channel further includes at least one middle mixing section between the third inlet anti the terminal mixing section; and the additional outlet of each of the at least one additional channel is connected to an intersection defined on the third channel and disposed between the terminal mixing section and one of the at least one middle mixing section or between two adjacent middle mixing sections when the third channel includes two or more middle mixing sections.

[0027] In some embodiments, the relative centrifugal force (RCF) is about 50xg to about 32,000xg. In some embodiments, the RCF is about 50xg to about l,000xg, about 50xg to about 3,000xg, about 50xg to about 5,000xg, about 50xg to about 10, 000x g, about 50xg to about I5,000xg, about 50xg to about 20,000xg, about 50xg to about 25,000xg, about 50xg to about 30,000xg, about 50xg to about 32,000xg, about 1 ,000xg to about 3,000xg, about 1 ,000xg to about 5,000xg, about L000xg to about l O.OOOxg, about l.OOOxg to about 15,000 xg, about 1.000xg to about 20.000xg, about l,000xg to about 25,000xgfabout l,000xg to about 30,000xg, about 1.000xg to about 32,000xg, about 3,000xg to about 5,000xg, about 3,000xg to about 10,000xg, about 3,000xg to about 15.000xg, about 3,000xg to about 20,000xg, about 3,000xg to about 25,000xg, about 3,000xg to about 30,000xg, about 3,000xg to about 32,000xg, about 5,000xg to about I0,000xg, about 5,000xg to about I5,000xg, about 5,000xg to about 20,000xg, about 5,000xg to about 25,000xg, about 5,000xg to about 30,000* g, about 5,000* g to about 32,000xg, about 10,000xg to about 15,000xg, about 10,000xg to about 20,(X)0xg, about 10,000xg to about 25,000xg, about 10,000xg to about 3(),000xg, about lO.OOOxg to about 32,000xg, about 15,000xg to about 20,000xg, about 15,000xg to about 25,000* g, about 15,000xg to about 30,000xg, about 15,000xg to about 32,0(X)xg, about 20,000xg to about 25,(X)0xg, about 20,000xg to about 30,000xg. about 20.000 ' g to about 32,OOOxg, about 25,000xg to about 30,000xg, about 25,000xg to about 32.000xg, or about 30,000xg to about 32,000 - g . In some embodiments, the RCF is about 50xg or more, about l,000xg or more, about 3,000xg or more, about 5,000xg or more, about lO.OOOxg or more, about 15,000xg or more, about 2Q,000xg or more, about 25,000xg or more, about 30,000 xg or more, or about 32,OOOxg or more. Preferably, the RCF is about 50xg or more, about 1 ,000xg or more, about 3,000xg or more, about 5,0()0xg or more, about 10,000xg or more, about 15,00()xg or more, about 20,00()xg or more, about 25,000xg or more, or about 30,000xg. In some embodiments, the RCF is about LOOOxg or less, about 3,000xg or less, about 5,000>;g or less, about 10,000 xg or less, about 15,000xg or less, about 20,000xg or less, about 25,0()0xg or less, about 30,000xg or less, or about 32,000xg or less.

[0028] In some embodiments, the device is centrifuged for I second (s) to 60 minutes (min), I s to 45 min, I s to 30 min, 1 s to 20 min, 1 s to 15 min, 1 s to 10 min, 1 s to 5 min, 1 s to 60 s, 1 s to 45 s, 1 s to 30 s, 1 s to 20 s, Is to 15 s, 1 s to 10 s, or 1 s to 5 s.

[0029] In some embodiments, the method is carried out at a controlled temperature. In some embodiments, the method is carried out by cooling or heating the device prior to centrifugation to temperatures from -20°C to 80”C. In some embodiments, the method is carried out by cooling or heating the one or more solutions prior to centrifugation to temperatures from -20°C io 80°C. In some embodiments, the method is carried out with the one or more solutions at different temperatures. In some embodiments, the centrifugation temperature is controlled from -20oC to 80°C. In some embodiments, the centrifugation temperature is 65°C.

[0030] In some embodiments, the device is centrifuged continuously. In some embodiments, two or more devices are centrifuged simultaneously to manufacture one or more distinct particles. In some embodiments, 24 to 96 devices are centrifuged simultaneously to manufacture one or more distinct particles. In some embodiments, multiple devices can be simultaneously centrifuged to produce multiple particles. In some embodiments, a device is centrifuged to manufacture one or more distinct particles.

[0031] In the present invention, ail solutions are in liquid form. In the present invention, the first solution, second solution, and additional solution may be a lipid solution, an aqueous phase solution, an organic phase solution, or any combination thereof.

[0032] In some embodiments, at least two solutions are introduced into the ports in fluid communication with the first intersection. In some embodiments, one of the at least two solutions is a therapeutic material, and the other is a solation comprising a particle-forming material or particles.

[0033] In some embodiments, the solution introduced into the port in fluid communication with the middle intersection or the terminal intersection may be a solution comprising a therapeutic material, a particle-forming material, particles, a targeting moiety, a solvent, a diluent, or any combination thereof.

[0034] In some embodiments, the solution comprises one or more therapeutic materials. In some embodiments, the solution comprises polynucleotide. In some embodiments, the solution is an aqueous solution. In some embodiments, the aqueous solution comprises a buffer solution. In some embodiments, the therapeutic material comprises a polynucleotide, a polypeptide, a protein, or any combination thereof. In some embodiments, the polynucleotide is a ribonucleic acid (RNA) fragment, a deoxyribonucleic acid (DNA) fragment, or a combination thereof. In some embodiments, the polynucleotide is an oligonucleotide. In some embodiments, the RNA comprises messenger RNA (mRNA), small interfering RNA (siRNA), transfer RNA ((RNA), ribosomal RNA (rRNA), small nuclear RNA (snRNA), small nucleolar RN A (snoRNA), micro RNA (miRNA), long noncoding RNA (IncRN A), circular RN A (circRNA), guide RNA (gRNA), and self-amplifying RNA (saRNA). In some embodiments, the DN A comprises double-stranded DNA (dsDNA), single- stranded DNA(ssDNA), plasmid DNA, linearized DNA, circular DNA, and minicircle DNA, In some embodiments, the therapeutic material is a small molecule.

[0035] In some embodiments, the solution comprises one or more particle- forming materials such as a lipid, a polymer, or a combination thereof. In some embodiments, the solution is in a liquid form, such as a lipid in ethanol or a polymer in acetonitrile. In some embodiments, the lipid comprises an ionizable cationic lipid, such as SM102, ALC-0315 and DLin-MO-DMA. In some embodiments, the polymer comprises a biocompatible polymer. In some embodiments, the polymer comprises poly(lactic-co-glycolic) acid (PLGA), In some embodiments, the solution is a combination of a lipid and a polymer, such as a hybrid of lipid and PLGA, which is used to manufacture hybrid PLGA / lipid nanoparticles.

[0036] In some embodiments, the solution comprises a solvent such as water, buffer solution, ethanol, methanol, isopropanol, chloroform, dichloroniethane, hexane, or acetonitrile. In some embodiments, the solution comprises one or more buffering agents such as Tris (tris(hydroxymethyl)aminomethane), Tris- HC1, sodium acetate, HEPES (4-(2-liydroxyethyl)-l -piperazineethanesulfonic add), or phosphate-buffered saline (PBS). In some embodiments, the solution pH is controlled from 4.5 to 8,5, In some embodiments, the solution comprises one or more stabilizing agents such as sucrose, trehalose, mannitol, sorbitol, or glycine. In some embodiments, the solution comprises one or more excipients such as ascorbic acid, EDTA (ethylenediaminetetraacetic acid), or citric acid.

[0037] In some embodiments, no solution is introduced into one or more ports in fluid communication with the middle intersection or the terminal intersection.

[0038] In the present invention, particle manufacturing is defined as particle formation, assembly, functionalization, loading, encapsulation (i.e. encapsulating therapeutic materials), modifications, or any combination thereof.

[0039] In some embodiments, the particles are nanoparticles or microparticles. In some embodiments, the particles are lipid-based nanoparticles or polymer- based nanoparticles. In some embodiments, the particles are liposomes. In some embodiments, the particles are lipid nanoparticles. in some embodiments, the particles are lipid microparticles or polymer microparticles. In some embodiments, the nanoparticles or microparticles encapsulate a therapeutic material. In some embodiments, the nanoparticle or microparticles are empty. In some embodiments, the nanoparticles encapsulate nucleic acid. In some embodiments, the lipid nanoparticles encapsulate nucleic acid. In some embodiments, the liposomes encapsulate nucleic acid. In some embodiments, the nanoparticles or microparticles are functionalized with one or more targeting moieties.

[0040] In some embodiments, the particles have a diameter of about 200 nm to

[0041] 1000 nm. In some embodiments, the nanoparticles have a diameter of about

[0042] 200 nm or less, as measured by dynamic light scattering. In some embodiments, the nanoparticles have a diameter of about 150 nm or less, as measured by dynamic light scatering. In some embodiments, the nanoparticles have a diameter of about 100 nm or less, as measured by dynamic light scattering. In some embodiments, the nanoparticles have a diameter of about 80 nm or less, as measured by dynamic light scattering. In some embodiments, the nanopartides have a diameter of about 50 nm or less, as measured by dynamic light scattering. In some embodiments, the nanoparticles have a diameter of about 30 nm or less, as measured by dynamic light scattering. In some embodiments, the microparticles have a diameter of about 1000 nm or less, as measured by dynamic light scattering. In some embodiments, the microparticles have a diameter of about 300 nm to about 800 nm, as measured by dynamic light scattering.

[0043] In some embodiments, the particles have a polydispersity index of about 0.3 or less, as measured by dynamic light scattering. In some embodiments, the particles have a polydispersity index of about 0.25 or less, as measured by dynamic light scatering. In some embodiments, the particles have a polydispersity index of about 0.2 or less, as measured by dynamic light scattering. In some embodiments, the particles have a polydispersity index of about 0.1.5 or less, as measured by dynamic light scattering. In some embodiments, the particles have a polydispersity index of about 0.1 or less, as measured by dynamic light scattering. In some embodiments, the particles have a poly dispersity index of about 0.1 to about 0.25, as measured by dynamic light scattering.

[0044] In some embodiments, the particles have an encapsulation efficiency of about 60% or more. In some embodiments, the particles have an encapsulation efficiency of about 70% or more. In some embodiments, the particles have an encapsulation efficiency of about 80% or more. I n some embodiments, the particles have an encapsulation efficiency of about 90% or more.

[0045] In some embodiments, the particles are spherical or non-spherical. In some embodiments, the particles comprise blebs. In some embodiments, the particles do not comprise blebs. In some embodiments, the particles exhibit a distinct core-shell structure. In some embodiments, the particles are multilamellar. In some embodiments, the particles exhibit a multi-layer structure.

[0046] In the present invention, the angle theta (0) is the angle between the principal flow direction and the centrifugal force direction. The principal flow direction is defined as the vector from the starting point to the ending point of a fluid path, irrespective of the actual path taken by the fluid. The principal flow direction represents the straight-line orientation of net fluid transport across a domain. The centrifugal force direction is defined as the outward radial direction from center of rotation. In some embodiments, the angle theta (0) is the angle between the principal flow direction and the centrifugal force direction. In some embodiments, the principal flow direction is parallel to the central axis of the device. In some embodiments, the channels are configured so that the principal flow direction is parallel to or angled within 9(f relative to the centrifugal force direction. In some embodiments, the device is positioned on the plane passing through the axis of rotation and the centrifugal force direction. In some embodiments, the angle theta (0) is 0° (degree) (i.e. principal flow direction is parallel to centrifugal force direction) to 90” (i .e. principal flow direction is perpendicular to centrifugal force direction). In some embodiments, the angle theta (0) is 45° . In some embodiments, the angle theta (0) is 30" .

[0047] In some embodiments, the one or more solutions converge at substantially the same time. In some embodiments, the one or more solutions enter the third channel sequentially. In some embodiments, the orientation of the device with respect to the centrifugal force direction dictates the one or more solutions enter the third channel sequentially or at substantially the same time. In some embodiments, the device is oriented in the centrifuge such that the principal flow direction is parallel to or angled within 90° relative to the centrifugal force direction. In some embodiments, the solution flow decelerates when flowing at a 90“ to 180° angle with respect to the centrifugal force direction. In some embodiments, the solution flow accelerates when flowing at a 0° to 90” angle with respect to the centrifugal force direction. In some embodiments, the entry timing into the third channel of the one or more solutions is controlled by the centrifugal force direction with respect to the device orientation. In some embodiments, the entry timing into the third channel of die one or more solutions is controlled by the relative centrifugal force. In some embodiments, the one or more solutions converge at substantially the same time under a relative centrifugal force of above 500xg.

[0048] In some embodiments, the device further comprises a third port for receiving a third solution, wherein the third port is in fluid communication with the third outlet of the third channel. In some embodiments, the method comprises introducing the third solution to the third port in fluid communication with the third outlet of the third channel. In some embodiments, the third solution comprises a solution for dilution such as water or an aqueous buffer. In some embodiments, the third solution comprises a solution comprising one or more stabilizing agents, such as sucrose, trehalose, mannitol, sorbitol, or glycine. In some embodiments, the third solution comprises a buffer solution with pH from 4.5 to 7.0. In some embodiments, the third solution comprises a buffer solution with pH from 7.0 to 8.5. In some embodiments, the method comprises introducing a third solution to a third port in fluid communication with the third outlet of the third channel prior to introducing solutions to the other ports. In sonic embodiments, the method comprises introducing a third solution to a third port in fluid communication with the third outlet to delay fluid flow in the device during centrifugation. In some embodiments, the third solution dilutes, stabilizes, or modifies the pH of the manufactured particles.

[0049] In some embodiments, the solution introduced into the corresponding port has a volume of about 20 microliters (pL) to about 1000 pL, 20 gJL to about 1000 uL, 20 pL to about 800 pL, 20 pL to about 600 uL, 20 pL to about 400 uL, 20 id to about 200 pL, 20 pL to about 180 uL, 20 pL to about 100 rd 20 uL to about 60 pL, 10 pl. to about 100 pL, 10 pL to about 50 uL, 5 pL to about 20 pL, 2 uL to about 10 uL, or 2 uL to about 5 pL. In some embodiments, the solution introduced into the corresponding port has a volume of about 1 milliliter (mL) to about 10 mL, 5 mL to about 10 mL, 1 mL to about 5 m l... or 1 mL to about 2 ml .. In some embodiments, the solution introduced into the corresponding port has a volume of about 10 mL or more. 15 ml. or more. 30 ml . or more, 50 mL or more, 100 mL or more, 1000 mL or more, or

[0050] 2000 mL or less.

[0051] In some embodiments, the volume ratio of the first solution and the second solution is between about 2: 1 and about 10: 1. In some embodiments, the volume ratio of the first solution and the second solution is about 2:1 , about 2.5: 1, about. 3:1, about. 3,5:1. about 4:1, about 4.5:1, about 5:1, about 5.5: 1, about 6: 1 , about 8:1, or about 10:1.

[0052] In some embodiments, the liquid height of each solution in the corresponding port is 5 millimeters (mm) to 150 mm, 5 mm to 120 mm, 5 ram to 100 mm, 5 mm to 80 mm, 5 mm to 60 mm, 5 mm to 40 mm, or 5 mm to 20 mm.

[0053] In some embodiments, the device is made of a thermoplastic such as poly(methyl methacrylate) (PMMA), polycarbonate (PC), cyclic olefin copolymer (COC), cyclic olefin polymer (COP), polypropylene (PP), or any combination thereof. In some embodiments, the device is made of glass or stainless steel. In some embodiments, the device is made of a biocompatible material such as a thermoplastic with surface treatment.

[0054] In the present invention, ports can play a role in the operation of a device, serving as the entry points through which fluids are introduced into the device or collection points for collecting resulting particles. These ports are designed to accept one or more fluids, which can then be manipulated within the device for various applications such as particle manufacturing, chemical processes, or fluid mixing. The design of these ports, in terms of their size, shape, and location, can influence the overall fluid dynamics within the device. For instance, they can be configured to control the liquid height, the relative distance of solutions to the center of rotation, liquid volume, ensuring the right amount of fluids are mixed at the appropriate time and at appropriate volumes to achieve desired outcomes.

[0055] In some embodiments, the device has 2 or more ports and corresponding channels. In some embodiments, the device has 3 or more ports and corresponding channels. In some embodiments, the device has 4 or more ports and corresponding channels. In some embodiments, the device has 5 or more ports and corresponding channels. In some embodiments, the device has 2 to 1536 ports and corresponding channels. In some embodiments, the device has 2 to 6 ports and corresponding channels. In some embodiments, the device has 2, 6, 12, 24, 48, 96, 384, or 1536 ports and corresponding channels, hi some embodiments, the device has 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 24, 36, 48, 96, 384, or 1536 ports and corresponding channels.

[0056] In the present invention, the solutions flow into the corresponding channels at different RCFs. In the present invention, the first solution flows through the first channel and the second solution flows through the second channel at different RCFs. In some embodiments, the first solution flows through the first channel, the second solution flows through the second channel, and the fourth solution flows through the fourth channel at different RCFs.

[0057] In the present invention, the solutions flow into the corresponding channels at substantially the same RCF. In the present invention, the first solution flows through the first channel and the second solution flows through the second channel at substantially the same RCF. In some embodiments, the first solution flows through the first channel, the second solution flows through the second channel, and the fourth solution flows through the fourth channel at substantially the same RCF.

[0058] In some embodiments, the device has multiple flow resistant sections with a resistance. In the present invention, the one or more resistances delay or selectively prevent fluid flow (such as the fluid flow of a solution) in a channel. In some embodiments, the one or more resistances control the timing of flow as to allow for full or partial contact of the solutions during centrifugation. In some embodiments, the resistance may prevent the fluid flow so that no fluid flow passes the section or region in the channel. In some embodiments, the one or more resistances individually control the flow timing in the corresponding channel under centrifugal force. In some embodiments, the one or more resistances facilitate flow convergence at substantially the same time under centrifugal force.

[0059] In some embodiments, the flow resistant sections are individually formed as a straight, circular, square, U-shaped, C-shaped, V-shaped, W- shaped, helical, serpentine or zig-zag channel. In some embodiments, the flow resistant section comprises one or more baffle structures. In some embodiments, the first How resistant section is formed as a straight, circular, square, U-shaped, C-shaped, V-shaped, W-shaped, helical, serpentine or zigzag channel. In some embodiments, the first flow resistant section comprises one or more baffle structures. In some embodiments, the second flow resistant section is formed as a straight, circular, square, U-shaped, C-shaped, V-shaped, W-shaped, helical, serpentine or zig-zag channel. In some embodiments, the second flow resistant section comprises one or more baffle structures. In some embodiments, the fourth, fifth and sixth flow resistant sections are individually formed as a straight, circular, square, U-shaped, C-shaped, V-shaped, W- shaped, helical, serpentine or zig-zag channel. In some embodiments, the fourth, fifth and sixth flow resistant sections comprise one or more baffle structures.

[0060] In some embodiments, the flow resistant sections individually have an inner diameter of about 10 μm or more. I n some embodiments, the flow resistant sections individually have an inner diameter of about 1 μm to about 1000 μm. In some embodiments, the flow resistant sections individually have an inner diameter of about 1 μm to about 6,000 μm. In some embodiments, the flow resistant sections individually have an inner diameter of about 1 μm to about 200 pm. about 1 μm to about 500 μm, about 1 μm to about 1 ,000 μm, about 1 μm to about 1,500 μm, about I μm to about 2,000 μm, about 1 p.ni to about 2,500 μm, about 1 μm to about 3.000 μm, about I μm to about 4,000 μm, about 1 μm to about 5,000 μm, about 1 μm to about 6,000 μm, about 100 pin to about 200 μm, about 100 μm to about 500 μm, about 100 μm to about 1 ,000 μm, about 100 μm to about 1,500 μm, about 100 μm to about 2,000 μm, about 100 μm to about 2,500 μm, about 100 μm to about 3,000 μm, about 100 μm to about 4,000 μm, about 100 μm to about 5,000 μm, about 100 μm to about 6,000 μm, about 200 μm to about 500 μm, about 200 μm to about 1 ,000 μm, about 200 jim to about 1 ,500 μm, about 200 μm to about 2,000 μm, about 200 μm to about 2,500 μm, about 200 μm to about 3,000 μm, about 200 μm to about 4,000 μm, about 200 μm to about 5,000 μm, about 200 μm to about 6,000 μm, about 500 μm to about 1,000 μm, about 500 μm to about 1 ,500 μm, about 500 jim to about 2,000 μm, about 500 pin to about 2,500 μm, about 500 μm to about 3,000 pm, about 500 μm to about 4,000 μm, about 500 p.m to about 5,000 μm, about 500 jam to about 6,000 μm, about 1,000 μm to about 1,500 μm, about 1,000 μm to about 2,000 μm, about 1,000 μm to about 2,500 μm, about 1,000 μm to about 3,000 μm, about 1 ,000 μm to about 4,000 μm, about 1,000 μm to about. 5,000 μm, about 1,000 μm to about. 6,000 μm, about 1,500 μm to about 2,000 μm, about 1,500 μm to about 2,500 μm, about 1 ,500 μm to about 3,000 μm, about 1 ,500 μm to about 4,000 μm, about 1,500 μm to about 5,000 μm, about 1 ,500 jtm to about 6,000 μm, about 2,000 μm to about 2,500 μm, about 2,000 μm to about 3,000 μm, about 2,000 μm to about 4,000 μm, about 2,000 μm to about 5,000 μm, about 2,000 μm to about 6,000 μm, about 2.500 μm to about. 3,000 pm, about 2,500 μm to about 4,000 μm, about 2,500 μm to about 5,000 jun, about 2,500 μm to about 6,000 μm, about 3,000 μm to about 4,000 μm, about 3,000 μm to about 5,000 μm. about 3,000 μm to about 6,000 μm, about 4,000 μm to about 5,000 jim, about 4.000 μm to about 6,000 jun, or about 5,000 μm to about 6,000 μm. In some embodiments, the flow resistant sections individually have an inner diameter of about 1 μm or more, about 100 μm or more, about 200 μm or more, about 500 μm or more, about 1,000 jun or more, about 1,500 jim or more, about 2,000 μm or more, about 2,500 μm or more, about 3,000 μm or more, about 4,000 jim or more, about 5,000 pm or more, or about 6,000 μm or more. In some embodiments, the flow resistant sections individually have an inner diameter of about 1 μm or more, about 100 μm or more, about 200 μm or more, about 500 pin or more, about 1 ,000 μm or more, about 1 ,500 μm or more, about 2,000 μm or more, about 2,500 μm or more, about 3,000 μm or more, about 4,000 μm or more, or about 5,000 μm or more. In some embodiments, the flow resistant sections individually have an inner diameter of about 100 μm or less, about 200 μm or less, about 500 μm or less, about 1,000 μm or less, about 1 ,500 μm or less, about 2,000 μm or less, about 2,500 μm or less, about 3,000 μm or less, about 4,000 |im or less, about 5,000 μm or less, or about 6,000 μm or less.

[0061] In the present invention, the term “hydraulic diameter” is a parameter in fluid dynamics, used extensively in characterizing the flow behavior within conduits such as pipes, channels. It provides a quantitative measure that can inform the behavior of fluid flow in non-circular conduits or complex-shaped channels. It is defined as four times the cross-sectional area of the flow divided by the wetted perimeter. In simpler terms, hydraulic diameter can be thought of as an equivalent diameter that represents the actual cross-section of a noncircular conduit where fluid is flowing. This parameter plays a crucial role in determining the flow resistance of a channel. Consequently, it operates as a component in designing and optimizing the performance of systems dealing with fluid flow for nano- and micro-particle manufacturing.

[0062] In some embodiments, the flow resistant sections individually comprise a hydraulic diameter between 1 μm (micrometer) and 10cm (centimeter), I μm and I cm, 1 μm and 5mm (millimeter), or 10μm and 5mm. In some embodiments, the first flow resistant section comprises a hydraulic diameter between 1 pm and 10cm, 1μm and 1cm, 1μm and 5mm, or 10μm and 5mm. In some embodiments, the second flow resistant section comprises a hydraulic diameter between 1μm andlOcm, 1μm and 1cm, 1 μm and 5mm, or 10μm and 5mm. In some embodiments, the terminal mixing section comprises a hydraulic diameter between I μm and 10cm, 1 μm and I cm, I μm and 5mm, or 10μm and 5mm. In some embodiments, the terminal mixing section comprises a hydraulic diameter between 100 μm and 800 μm, 100 μm and 600 μm, 100 μm and 500 μm, 100 μm and 300 μm, or 100 μm and 200 μm. In some embodiments, each of the at least one middle mixing section comprises a hydraulic diameter between 1 μm and 10cm, 1μm and 1cm, 1μm and 5 mm , or 10μm and 5mm. In some embodiments, the fourth, fifth and sixth flow resistant sections individually comprise a hydraulic diameter between 1 μm and 10cm, 1 μm and 1cm, 1 μm and 5mm, or 10μm and 5mm.

[0063] In some embodiments, the device may have multiple intersections. In some embodiments, each of the intersections is a Y junction, a T junction, or a junction of n-way. In some embodiments, the first intersection is a Y junction, a T junction, or a junction of n-way. In some embodiments, the terminal intersection is a Y junction, a T junction, or a junction of n-way. In some embodiments, each of the middle intersections is a Y junction, a T junction, or a junction of n-way.

[0064] Upon centrifugation beyond a certain RCF (i.e. greater than i xg-force), the solutions may flow and interact with, each other through a series of connected channels. These channels can consist of junction of n-ways, flow focusing channels, helical, serpentine, zig-zag channels with one or more variable cross sections such as baffle structures, or bifurcating structures to induce cross-flow, chaotic advection, relaminating flow, or Dean vortices, resulting in increased mixing efficiency for manufacturing nanoparticles or in i cro partic les.

[0065] In some embodiments, the flow resistant sections individually comprise one or more passive valves. In some embodiments, the first flow resistant section comprises one or more passive valves. In some embodiments, the second flow resistant section comprises one or more passive valves. In some embodiments, the terminal mixing section comprises one or more passive valves. In some embodiments, each of the at least one middle mixing section comprises one or more passive valves.

[0066] In some embodiments, the passive valve has a hydraulic diameter between about 1 μm and about 1 cm. In some embodiments, the passive valve has a hydraulic diameter between about 10 μm and about 5 mm. In some embodiments, the passive valve has a hydraulic diameter between about I p.m and about 6,000 μm. In some embodiments, the passive valve has a hydraulic diameter between about 1 μm and about 100 μm, about 1 μm and about 200 urn, about I uni and about 500 μm, about 1 μm and about 1 ,000 μm, about 1 μm and about 1 ,500 μm, about I μm and about 2,000 μm, about 1 μm and about 2,500 μm, about 1 μm and about 3,000 μm, about 1 μm and about 4,000 μm, about 1 μm and about 5,000 μm, about 1 μm and about 6,000 μm, about 100 μm and about 200 μm, about 100 μm and about 500 μm, about 100 μm and about 1,000 μm, about 100 μm and about 1 ,500 μm, about 100 μm and about 2.000 μm, about 100 μm and about 2.500 μm, about 100 μm and about 3,000 pin, about 100 uni and about 4,000 μm, about 100 μm and about 5,000 μm, about 100 μm and about 6.000 μm, about 200 μm and about 500 μm, about 200 μm find about 1,000 μm, about 200 μm and about 1 ,500 μm, about 200 μm and about 2,000 μm, about 200 μm and about 2,500 μm, about 200 μm and about 3,000 pm, about 200 μm and about 4,000 μm, about 200 μm and about 5,000 μm, about 200 μm and about 6,000 μm, about 500 μm and about 1,000 μm, about 500 μm and about 1,500 μm, about 500 μm and about 2,000 μm, about 500 μm and about 2,500 μm, about 500 μm and about 3,000 μm, about 500 μm and about 4,000 μm, about 500 μm and about 5,000 μm, about 500 μm and about 6,000 pm, about 1,000 μm and about 1,500 μm, about 1,000 μm and about 2,000 μm, about 1,000 μm and about 2,500 μm, about 1,000 μm and about 3,000 μm, about 1 ,000 μm and about 4,000 μm, about 1,000 μm and about 5,000 μm, about 1 ,000 μm and about 6,000 μm, about 1,500 μm and about 2,000 μm, about 1,500 μm and about 2,500 μm, about 1,500 μm and about 3,000 μm. about 1,500 μm and about 4,000 μm, about 1 ,500 μm and about 5,000 μm, about 1 ,500 μm and about 6,000 μm, about 2,000 μm and about 2,500 μm, about 2,000 μm and about 3,000 μm, about 2,000 gm and about 4,000 μm, about 2,000 μm and about 5,000 μm. about 2,000 gin and about 6,000 μm, about 2,500 μm and about 3,000 μm, about 2,500 μm and about 4,000 μm, about 2,500 μm and about 5,000 μm, about 2,500 μm and about 6,000 μm, about 3.000 μm and about 4,000 μm, about 3,000 gm and about 5,000 μm, about 3,000 μm and about 6,000 μm, about 4,000 gm and about 5,000 μm, about 4,000 μm and about 6,000 μm, or about 5,000 μm and about 6,000 μm. In some embodiments, the passive valve has a hydraulic diameter of about 1 μm, about 100 μm, about 200 μm, about 500 μm, about 1 ,000 μm, about 1 ,500 μm. about 2.000 μm, about 2,500 μm, about 3,000 μm, about 4,000 μm, about 5,000 μm, or about 6,000 μm. In some embodiments, the passive valve has a hydraulic diameter of about 1 μm or more, about 100 μm or more, about 200 μm or more, about 500 μm or more, about 1,000 pm or more, about 1 ,500 μm or more, about 2,000 μm or more, about 2,500 μm or more, about 3.000 μm or more, about 4,000 μm or more, or about 5,000 μm or more. In some embodiments, the passive valve has a hydraulic diameter of about 100 μm or less, about 200 μm or less, about 500 μm or less, about 1,000 μm or less, about 1,500 μm or less, about 2,000 μm or less, about 2,500 μm or less, about 3,000 μm or less, about 4,000 μm or less, about 5,000 μm or less, or about 6,000 μm or less.

[0067] In some embodiments, the device further comprises one or more filtering parts, In some embodiments, the filtering part comprises one or more filter membranes. In some embodiments, the filtering part comprises frits. In some embodiments, the filtering part comprises size-exclusion beads. In some embodiments, the filtering part is a cellular membrane (such as PTFE, PVDF, PES, and cellulose acetate). In some embodiments, the filtering part is disposed below the third outlet to filter the particles. In some embodiments, each port or channel outlet comprises a filtering part.

[0068] In some embodiments, the kit further comprises an outer container. In some embodiments, the outer container collects particles from the device. In some embodiments, the outer container is a centrifuge tube. In some embodiments, the container is a centrifuge tube of about 0.2 ml.. or more volume capacity. In some embodiments, the outer container is a centrifuge tube of about

[0069] 0.2 ml . 0.5 mL. I ml, 1.5 mL, 1.8 ml, 2 mL, 3 mL, or 5 mL. In some embodiments, the outer container is a centrifuge tube of about 1 mL, 50 mL, 100 mL, 500 mL, or 1000 mL capacity.

[0070] In some embodiments, the outer container is a well plate. In some embodiments, the well plate fits American National Standards Institute, Inc. / Society for Laboratory Automation and Screening (ANS1 / SLAS) standard. In some embodiments, the well plate is a non-standard well plate comprising two or more wells, and less than 192 wells. In some embodiments, the outer container is a well plate comprising 6 wells, 12 wells, 24 wells, 48 wells, or 96 wells, hi some embodiments, the standard well plate is a 6-well plate, a 12-well plate, a 24-well plate, a 48-weil plate, or a 96-well plate. In some embodiments, the outer container is a deep well plate or a filter well plate.

[0071] In some embodiments, two or more devices are arranged in a strip well format compatible with well plates. In some embodiments, 2, 4, 8, or 12 devices are arranged in a strip well format compatible with well plates. In some embodiments, 12 to 96 devices are arranged in a strip well format compatible with well plates.

[0072] In some embodiments, the system for manufacturing particles comprises a device in the present invention and a centrifuge. In some embodiments, the system for manufacturing particles comprises a device, a container, and a centrifuge. In some embodiments, the system comprises the kit and a centrifuge. In some embodiments, the centrifuge comprises a fixed-angle rotor or swing-out rotor. In some embodiments, automation is applied to the system. In some embodiments, robotic handling is used.

[0073] Except in the examples, or where otherwise expressly indicated, all numerical quantities in this description indicating amounts of material or conditions of process and / or use are to be understood as modified by the word ''about" in describing the broadest scope of the invention. Practice within the numerical limits stated is generally preferred. Also, unless expressly staled to the contrary the term "polymer" includes "oligomer,'' "copolymer," "terpolymer," and the like; molecular weights provided for any polymers refer to weight average molecular weight unless otherwise indicated; the description of a group or class of materials as suitable or preferred for a given purpose in connection with the invention implies that mixtures of any two or more of the members of the group or class are equally suitable or pre ferred; description of constituents in chemical terms refers to the constituents at the time of addition to any combination specified in the description, and does not necessarily preclude chemical interactions among the constituents of a mixture once mixed; the first definition of an acronym or other abbreviation applies to all subsequent uses herein of the same abbreviation and applies mutatis mutandis to normal grammatical variations of the initially defined abbreviation; arid, unless expressly stated to the contrary, measurement of a property is determined by the same technique as previously or later referenced for the same property.

[0074] It is also to be understood that this invention is not limited to the specific embodiments and methods described below, as specific components and / or conditions may. of course, vary. Furthermore, the terminology used herein is used only for the purpose of describing particular embodiments of the present invention and is not intended to be limiting in any way.

[0075] It must also be noted that, as used in the specification and the appended claims, the singular forms ’’a,” "an,” and "the" comprise plural referents unless the context clearly indicates otherwise. For example, reference to a component in the singular is intended to comprise a plurality of the components.

[0076] The term “comprising” is synonymous with “including,” “having,” “containing,” or “characterized by.” These terms are inclusive and open-ended and do not exclude additional, unrecited elements or method steps.

[0077] The phrase “consisting of* excludes any element, step, or ingredient not specified in the claim. When this phrase appears in a clause of the body of a claim, rather than immediately following the preamble, it limits only the element set forth in that clause; other elements are not excluded from the claim as a whole.

[0078] The phrase “consisting essentially of” limits the scope of a claim to the specified materials or steps, plus those that do not materially affect the basic and novel characteristic! s) of the claimed subject matter.

[0079] With respect to the terms “comprising,” “consisting of,” and “consisting essentially of,” where one of these three terms is used herein, the presently disclosed and claimed subject matter can include the use of either of the other two terms.

[0080] It should also be appreciated that integer ranges explicitly include all intervening integers. For example, the integer range 1-10 explicitly includes 1 , 2, 3, 4, 5, 6, 7, 8, 9, and 10. Similarly, the range 1 to 100 includes 1, 2, 3, 4. . . .

[0081] 97, 98, 99, 100. Similarly, when any range is called for, intervening numbers that are increments of the difference between the upper limit and the lower limit divided by 10 can be taken as alternative upper or lower limits. For example, if the range is 1.1. to 2.1 the following numbers 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9. and 2.0 can be selected as lower or upper limits. In the specific examples sei forth herein, concentrations, temperature, and process conditions (e.g. RCF, pH, volume ratio etc. ) can be practiced with plus or minus 50 percent of the values indicated rounded to three significant figures. In a refinement, concentrations, temperature, and process conditions (e.g., RCF, pH, volume ratio etc.) can be practiced with plus or minus 30 percent of the values indicated rounded to three significant figures of the value provided in the examples. In another refinement, concentrations, temperature, and process conditions (e.g., RCF, pH, volume ratio etc.) can be practiced with plus or m inus 10 percent of the values indicated rounded to three significant figures of the value provided in the examples.

[0082] In the examples set forth herein, concentrations, temperature, and process conditions (e.g,, RCF, pH, volume ratio, etc.) can be practiced with plus or minus 50 percent of the values indicated rounded to or truncated to two significant figures of the value provided in the examples. In a refinement, concentrations, temperature, and process conditions (e.g., RCF, pH, volume ratio, etc.) can be practiced with plus or minus 30 percent of the values indicated rounded to or truncated to two significant figures of the value provided in the examples. In another refinement, concentrations, temperature, and process conditions (e.g., RCF, pl l. volume ratio, etc.) can be practiced with plus or minus 10 percent of the values indicated rounded to or truncated to two significant figures of the value provided in the examples.

[0083] The term “fluid communication” refers to the ability of fluid to move from one part, element, or component to another; or the state of being connected, such that fluid can move from one portion that is connected to another portion.

[0084] The term “particle manufacturing” is defined as particle formation, assembly, functionalization, loading, encapsulation (i.e. encapsulating therapeutic materials), modifications, or any combination thereof.

[0085] The term “nano” means that at least one dimension is at most 500 nm. In a refinement, “nano’’ means that at least one dimension is at most 200 nm.

[0086] The term "solution" refers to any fluidic specimen introduced into the device or system for analysis or processing. These solutions can span a wide variety of substances and can come from numerous sources. For instance, they might be biological fluids such as blood, urine, or saliva used in medical testing. They could also be environmental solutions like water or air, tested for pollution levels. In pharmaceutical or chemical processes, solutions might be gasses, solutions of chemicals, drugs, or other substances undergoing processing, reaction, synthesis, or analysis. Therefore, a solution in this context is any fluidic material that is subject to investigation or manipulation within a fluidic system.

[0087] Throughout this application, where publications are referenced, the disclosures of these publications in their entireties are hereby incorporated by reference into this application to more fully describe the state of the art to which this invention pertains.

[0088] BRIEF DESCRIPTION OF THE DRAWINGS

[0089] FIG. 1 is a schematic view of the device of the present invention.

[0090] FIG. 2 is another schematic view of the device of the present invention.

[0091] FIG, 3 is another schematic view of the device of the present invention.

[0092] FIG. 4 is another schematic view of the device of the present invention.

[0093] FIG. 5 is another schematic view of the device of the present invention,

[0094] FIG. 6 is another schematic view of the device of the present invention.

[0095] FIG. 7 is another schematic view of the device of the present invention.

[0096] FIG. 8 is another schematic view of the device of the present invention.

[0097] FIG. 9 is a schematic view of setting the device of the present invention in a centrifuge tube.

[0098] FIG. 10 is a schematic view of placing the centrifuge tube with the device into a centrifuge for centrifugation.

[0099] FIG. Il is a schematic view of collecting the resulting particles in the centrifuge tube after centrifugation.

[0100] FIG. 12 is a schematic view of the relationship of the principal flow direction that is parallel to the central axis of the device and the centrifugal force direction on the plane passing through the axis of rotation and the central axis of the device.

[0101] FIG. 13 is a schematie view of setting the device of the present invention in a well-plate for centrifugation.

[0102] FIG. 14 shows the results of panicle size distribution of Preparation Example 1 (PEI) and Comparative Preparation Example 1 (CPE1 ).

[0103] FIG. 15 shows the results of particle size and polydispersity index (PDI) of PEl and CPE1.

[0104] FIG. 16 shows the results of transfection efficiency of PEI and CPE1.

[0105] FIG. 17 shows the results of particle size and polydispersity index of Preparation Example 2 (PE2) and Preparation Example 3 (PE3).

[0106] FIG. 18 shows the results of transfection efficiency of PE2 and PE3.

[0107] FIG. 19 shows the results of particle size distribution of Preparation Example 3 (PE3), Preparation Example 4 (PE4) and Preparation Example 5 (PE5).

[0108] FIG. 20 shows the results of particle size and polydispersity index of PE3. PE4 and PES.

[0109] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0110] With reference to FIG. 1, the device 1 of the present invention comprises a first port A, a first channel 10, a second port B, a second channel 20. and a third channel 30. The first port A is for receiving a first solution. The first channel 10 comprises a first inlet and a first outlet, and the first in let is in fluid communication with the first port A. The second port B is for receiving a second solution. The second channel 20 comprises a second inlet and a second outlet. The second inlet is in fluid communication with the second port B. The third channel 30 comprises a third inlet and a third outlet. The device 1 farther comprises a first intersection IS- 1 of the first outlet, the second outlet and the third inlet, such that the third inlet is in fluid communication with the first outlet and the second outlet. The first solution in the first port A flows through the first channel 10 and the second solution in the second port B flows through the second channel 20 when a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device 1.

[0111] With reference to FIG. 2, the device 1 of the present invention is similar to FIG. 1, but further comprises a first flow resistant section 11 and a second flow resistant section 21. The first channel 10 comprises a first flow resistant section 11 disposed between the first, inlet, and the first outlet and is configured to control fluid flow in the first channel 10. The second channel 20 comprises a second flow resistant section 21 disposed between the second inlet and the second outlet and is configured to control fluid flow in the second channel 20. The third channel 30 comprises a third inlet, a third outlet, and a terminal mixing section 31. The terminal mixing section 31 is disposed between the third inlet and the third outlet and is configured to mix fluid flow in the third channel 30 and form particles. The first solution in the first port A flows through the first, channel 10 and the second solution in the second port B flows through the second channel 20 when a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device 1.

[0112] With reference to FIG. 3, the device 1 of the present invention is similar to FIG. 2, but further comprises a third port C disposed between the terminal mixing section 31 and the third outlet.

[0113] When any one of the devices 1 of FIGs. 1 to 3 is provided, a first solution is introduced into the first port A, and a second solution is introduced into the second port B; and, a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device I to make the first solution pass through the first channel 10 and the second solution pass through the second channel 20; the first solution and the second solution contact at the first intersection IS- 1 and mix at the terminal mixing section 31 to form a first fluid comprising first particles comprising the first solution and the second solution.

[0114] With reference to FIG. 4, the device 1 of the present invention is similar to FIG. 2, but further comprises a fourth port D and a fourth channel 40. The fourth port D is for receiving a fourth solution. The fourth channel 40 comprises a fourth inlet, a fourth outlet, and a fourth flow resistant section 41 . The fourth inlet is in fluid comm uni cation with the fourth port D. The fourth outlet is connected to the first intersection IS- 1 and in fluid communication with the third channel. The fourth flow resistant section 41 is disposed between the fourth inlet and the fourth outlet and is configured to control fluid flow in the fourth channel 40. The first solution in the first port A flows through the first channel 10, the second solution in the second port B flows through the second channel 20, and the fourth solution in the fourth port D flow s through the fourth channel 40 when a relative centrifugal force (R.CF) of 5()xg to 32000xg is applied to the device I.

[0115] When the device 1 of FIG. 4 is provided, a first solution is introduced into the first port A, a second solution is introduced into the second port B, and a fourth solution is introduced into the fourth port D; and, a relative centrifugal force (RCF) of 50xg to 32000 xg is applied to the device 1 to make the first solution pass through the first channel 10, the second solution pass through the second channel 20, and the fourth solution pass through the fourth channel 40; the first solution, the second solution and the fourth solution contact at the first intersection IS- 1 and mix at the terminal mixing section 31 to form a second fluid comprising second particles comprising the first solution, the second solution and the fourth solution.

[0116] With reference to FIG. 5, the device 1 of the present invention is similar to FIG. 2, but further comprises a middle mixing section 32, a fifth port E and a fifth channel 50. The fifth port E is for receiving a fifth solution. The fifth channel 50 comprises a fifth inlet, a fifth outlet, and a fifth flow resistant section 51 . The fifth inlet is in fluid communication with the fifth port E. The fifth outlet is connected to a terminal intersection IS-T and in fluid communication with the third channel 30, The fifth flow resistant section 51 is disposed between the fifth inlet and the fifth outlet and is configured to control fluid flow in the fifth channel 50, The first solution in the first port A flows through the first channel 10, the second solution in the second port B flows through the second channel 20, and the fifth solution in the fifth port E flows through the fourth channel 50 when a relative centrifugal force (RCF) of 5(fog to 32000xg is applied to the device 1.

[0117] When the device 1 of FIG. 5 is provided, a first solution is introduced into the first port A, a second solution is introduced into the second port B, and a fifth solution is introduced into the fifth port E; and, a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device 1 to make the first solution pass through the first channel 10, the second solution pass through the second channel 20, and the fifth solution pass through the fifth channel 50; the first solution and the second solution contact at the first intersection IS-1 and mix at the middle mixing section 32 to form a first fluid comprising first particles comprising the first solution and the second solution; and the first fluid and the fifth solution contact at the terminal intersection IS-T and mix at the terminal mixing section. 31 to form a third fluid comprising third particles. The third particles can be the same as the first particles; or, the third particles can be the first particles modified by the fifth solution.

[0118] With reference to F IG. 6, the device 1 of the present invention is similar to FIG. 4, but further comprises a middle mixing section 32, a fifth port E and a fifth channel 50. The fifth port E is for receiving a fifth solution. The fifth channel 50 comprises a fifth inlet, a fifth outlet, and a fifth flow resistant section 51. The fifth inlet is in fluid communication with the fifth port E. The fifth outlet is connected to the terminal intersection IS-T and in fluid communication with the third channel. The fifth flow resistant, section 51 is disposed between the fifth inlet and the fifth outlet and is configured to control fluid flow in the fifth channel 50. The first solution in the first port A flow's through the first channel 10, the second solution in the second port B flows through the second channel 20, the fourth solution in the fourth port D flows through the fourth channel 40, and the fifth so hition in the fifth port E flows through the fourth channel 50 when a relative centrifugal force (RCF) of 50 vg to 32000 x g is applied to the device 1 .

[0119] When the device 1 of FIG. 6 is provided, a first solution is introduced into the first port A, a second solution is introduced into the second port B, a fourth solution is introduced into the fourth port D, and a fifth solution is introduced into the fifth port E; and, a relative centrifugal force (RCF) of 50 xg to 32000 xg is applied to the device 1 to make the first solution pass through the first channel 10, the second solution pass through the second channel 20, the fourth solution pass through the fourth channel 40, and the fifth solution pass through the fifth channel 50: the first solution, the second solution and the fourth solution contact at the first intersection IS-l and mix at the middle mixing section 32 to form a second fluid comprising second particles comprising the first solution, the second solution and the fourth solution; and the second fluid and the fifth solution contact at the terminal intersection IS-T and mix at the terminal mixing section 31 to form a fourth fluid comprising fourth particles. The fourth particles can be the same as the second particles; or, the fourth particles can be the second particles modified by the fifth solution.

[0120] With reference to FIG. 7, the device 1 of the present invention is similar to FIG, 5, but further comprises another middle mixing section 32, a sixth port F and a sixth channel 60, and the fifth outlet is connected to a middle intersection IS-M and in fluid communication with the third channel. The sixth port F is for receiving a sixth solution. The sixth channel 60 comprises a sixth inlet, a sixth outlet, and a sixth flow resistant section 61. The sixth inlet is in fluid communication with the sixth port E The sixth outlet is connected to the terminal intersection IS-T and in fluid communication with the third channel. The sixth flow resistant section 61 is disposed between the sixth inlet and the sixth outlet and is configured to control fluid flow in the sixth channel 60. The first solution in the first port A flows through the first channel 10, the second solution in the second port B flows through the second channel 20, the fifth solution in the fifth port E flows through the fourth channel 50, and the sixth solution in the fourth port F flows through the sixth channel 60 when a relative centrifugal force (RCF) of 50xg to 32000 xg is applied to the device I.

[0121] When the device I of FIG. 7 is provided, a first solution is introduced into the first port A, a second solution is introduced into the second port B, a fifth solution is introduced into the fifth port E, and a sixth solution is introduced into the sixth port F; and, a relative centrifugal force (RCF) of 50xg to 32000 xg is applied to the device 1 to make the first solution pass through the first channel 10, the second solution pass through the second channel 20, the fifth solution pass through the fifth channel 50, and the sixth solution pass through the sixth channel 60; the first solution and the second solution contact at the first intersection IS- 1 and mix at the middle mixing section 32 to form a first fluid comprising first particles comprising the first solution and the second solution; the first fluid and the fifth solution contact at the middle intersection IS-M and mix at the other middle mixing section 32 to form a third fluid comprising third particles; and the third fluid and the sixth solution contact at the terminal intersection IS-T and mix at the terminal mixing section 31 to form a fifth fluid comprising fifth particles. The fifth particles can be the same as the third particles; or, the fifth particles can be the third particles modified by the six solation. In addition, the third particles can be the same as the first particles; or, the third particles can be the first particles modified by the fifth solution.

[0122] With reference to FIG. 8, the device 1 of the present invention is similar to FIG, 6, but further comprises another middle mixing section 32, a sixth port F and a sixth channel 60, and the fifth outlet is connected to the middle intersection IS-M and in fluid communication with the third channel. The sixth port F is for receiving a sixth solution. The sixth channel 60 comprises a sixth inlet, a sixth outlet, and a sixth flow resistant section 61. The sixth inlet is in fluid communication with the sixth port F. The sixth outlet is connected to the terminal intersection IS-T and in fluid communication with the third channel. The sixth flow resistant section 61 is disposed between the sixth inlet and the sixth outlet and is configured to control fluid flow in the sixth channel 60. The first solution in the first port A flows through the first channel 10, and the second solution in the second port B flows through the second channel 20, the fourth solution in the fourth port D flows through the fourth channel 40, the fifth solution in the fifth port E flows through the fourth channel 50, and the sixth solution in the sixth port F flows through the sixth channel 60 when a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device 1.

[0123] When the device 1 of FIG. 8 is provided, a first solution is introduced into the first port A, a second solution is introduced into the second port B, a fourth solution is introduced into the fourth port D, a fifth solution is introduced into the fifth port E, and a sixth solution is introduced into the sixth port F; and, a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device I to make the first solution pass through the first channel 10, the second solution pass through the second channel 20, the fourth solution pass through the fourth channel 40, the fifth solution pass through the fifth channel 50, and the sixth solution pass through the sixth channel 60; the first solution, the second solution and the fourth soltition contact at the first intersection IS- 1 and mix at the middle mixing section 32 to form a second fluid comprising second particles comprising the first solution, the second solution and the fourth solution; and the second fluid and the fifth solution contact at the middle intersection IS-M and mix at the other middle mixing section 32 to tom a fourth fluid comprising fourth particles; and the fourth fluid and the sixth solution contact at the terminal intersection IS-T and mix at the terminal mixing section 31 to form a sixth fluid comprising sixth particles.

[0124] With reference to FIGs. 9 to 13, a device 1 lor manufacturing particles in accordance with the present invention is configured to be set in an outer container. As shown in FIGs. 9 and 13, the outer container may be a centrifuge tube 71 or a well plate 72. The centrifuge tube 71 may be a microcentrifiige tube having a size of 1.5 milliliters (mL) or 2 mL, 5 m.L, or a centrifuge tube having a size of 15 mL or 50 mL. The well plate 72 fits ANSI / SLAS standard and may be a 6-well plate, a 12-well plate, a 24-weil plate, a 48-well plate, or a 96-well plate.

[0125] With reference to FIG.12, on the plane passing through the axis of rotation and the central axis the device I , the principal flow direction (indicated by the arrow with a dash-dotted line) forms an angle 0 with the centrifugal force direction (indicated by the arrow with a solid line).

[0126] The timing of flow is controlled to facilitate the convergence of solutions at substantially the same time. Specifically, the first flow resistant section 11 , the second flow resistant section 21 , the terminal mixing section 31 , the middle mixing section 32, the fourth flow resistant section 41 , the fifth flow resistant section 51 and the sixth flow resistant section 61 may be formed as a straight, circular, square, U-shaped. C-shaped, V-shaped, W-shaped, helical. serpentine or zig-zag channel and may comprise a hydraulic diameter between 10μm (micrometer) and 1000 μm.

[0127] By selectively preventing the fluid flow, the timing of flow is also able to be controlled. Specifically, the first flow resistant section 11 , the second flow resistant section 21, the fourth flow resistant section 41, the fifth flow resistant section 51 and the sixth flow resistant section 61 may be a passive valve.

[0128] As shown in FIGs. 9 to 11, the solutions are introduced into the corresponding ports of the device and the device I is set in a centrifuge tube 71, the centrifuge tube 71 with the device 1 is placed into a centrifuge CM for centrifugation, and the resulting particles can be collected in the centrifuge tube 71. Solutions may be introduced before or after setting the device 1 in the centrifuge tube 71.

[0129] As shown in FIG. 13, the device 1 can be set in a well-plate 72 for centrifugation. The centrifuge CM can use a fixed-angle or swing-out rotor.

[0130] Preparation Example 1: mRNA lipid nanoparticles (mRNA-LNPs) prepared by the device of the present Invention

[0131] In this example, the device as shown in FIG. 2 was used to prepare mRNA lipid nanoparticles by using the solutions below.

[0132] First solution: 200 uL an aqueous mRNA solution with a concentration of 20 nanograms per microliter (ng / pL) in 25 mM sodium acetate (pH 5.2). The mRNA is the Firefly- Luciferase (F-Luc) mRNA purchased from GenScript.

[0133] Second solution: 40 |.iL of mixture containing 50 mol% of SMI 02 (also named 9-Heptadecanyl 8- {(2-hydroxyethyl)[6-oxo-6- (undecyloxy)bexyl]ami no} octanoate), 10 mol% DSPC (also named 1,2- dioctadecanoyl-sn-glycero-3-phosphocholine), 38.5 mol% cholesterol, 1.5 moi% DMG-PEG2000(also named 1 ,2<iimyristoyl-rac-glycero-3- meth oxypolyethylene giycoi-2000) with a combined lipid concentration of 3.6 millimolar (mM) in ethanol. The lipids were purchased from Cayman Chemical.

[0134] The device of the present invention was set in a centrifuge tube. The first solution was added into the first port. The second solution was added into the second port. The centrifuge tube with the device was placed into a centrifuge with a 45-degree fixed angle rotor (0 = 45”). The centrifuge was operated with a RCF of 2000xg for 30 seconds to obtain a solution comprising mRNA lipid nanoparticles (mRNA -LNPs). The centrifuge tube was removed from the centrifuge and the device was removed from the centrifuge tube. The resulting mixture comprising the nanoparticles at the bottom of the centrifuge tube was removed and characterized.

[0135] Preparation Example 2: mRNA lipid nanoparticles (mRNA -LNPs) prepared by the device of the present Invention

[0136] The mRNA lipid nanoparticles were prepared with the method described in Preparation Example 1, except the first solution is 180 itL an aqueous mRNA solution with a concentration of 46 ng / uL, the second solution is 60 pL lipid mixture with a concentration of 5 mM in ethanol, and the volume ratio of port A to port B is 3, and the resistance ratio of the first flow resistant section to the second How resistant section is about 2.5 to 3.5.

[0137] Preparation Example 3: mRNA lipid nanoparticles (mRNA -LNPs) prepared by the device of the present invention The mRNA lipid nanoparticles were prepared with the method described in Preparation Example 1, except the first solution is 200 uL an aqueous mRNA solution with a concentration of 28.3 ng / 'pL, the second solution is 40 pL SM I 02 with a concentration of 5 mM in ethanol, and the volume ratio of port A to port B is 5, and the resistance ratio of the first flow resistant section to the second flow resistant section is about 4 to 6.

[0138] Preparation Example 4: mRNA lipid nanoparticles (mRNA -LNPs) prepared by the device of the present invention

[0139] The mRNA lipid nanoparticles were prepared with the method described in Preparation Example k except the first solution is 200 pL an aqueous mRNA solution with a concentration of 26.2 ng / pL, the second solution is 40 pL a mixture containing 46,3 mol% of ALC-0315 (also named [(4-Hydroxybutyl)azanediyl]di(hexane-6, 1 -diyl) bis(2-hexyidecanoate)), 9,4 mol% DSPC (also named l,2-dioctadecanoyl-sn-glycero-3-phosphocholine), 42.7 mol% Cholesterol, 1.6 mol% ALC-0159 (also named

[0140] Methoxy po ly ethy leneg lycoloxyi 2000 )-N , N -ditetradecy I acetam ide ) with a combined lipid concentration of 5 mM in ethanol, and the volume ratio of port A to port B is 5. The lipids were purchased from Cayman Chemical.

[0141] Preparation Example 5: mRNA lipid nanoparticles (mRNA -LNPs) prepared by the device of the present invention

[0142] The mRNA lipid nanoparticles were prepared with the method described in Preparation Example 1, except the first solution is 200 uL an aqueous mRNA solution with a concentration of 56.6 ng / pL. the second solution is 40 pL of mixture containing 50 mol% of DLin-MC3-DM A (also named 4-(dimethylamino)-butanoic acid, ( 10Z, 13Z)-1 -(9Z, 12Z)-9,12- octadecadien-l-yl-10,13-nonadecadien-l-yl ester), 10 mo1% DS PC (also named 1 ,2-dioetadecanoyl-sn-glycero3'phosphocholine), 38.5 1110!% Cholesterol, 1.5 mol% DMG-PEG2000(also named 1 ,2-dimyristoyl-rac- glyccro-3-metlioxypolyethylene gIycoi-2000) with a combined lipid concentration of 5 mM in ethanol, and the volume ratio of port A to port B is 5. The lipids were purchased from Cayman Chemical.

[0143] Comparative Preparation Example 1: mRNA lipid nanoparticles (mRNA -LNPs) prepared by pipette mixing

[0144] With the same first and second solutions used in Preparation Example 1, niR.NA lipid nanoparticles were also formed using pipette mixing (a manual mixing method, see Nature Protocols, volume 18, pages 265—291 (2023)).

[0145] Testing Example 1: Analysis of physicochemical characteristics and encapsulation efficiency

[0146] The size and polydispersity of the mRNA -LNPs obtained in Preparation Examples 1 to 5 (PEI to PE5) and Comparative Preparation Example 1 (CPE I) were directly analyzed by a dynamic light scattering (DCS) analyzers of Litesizef™ 500 (Anton Paar) with a refractive index of 1.45 and Cumulant Model ISO 22412 , the standard method for DLS analysis is ISO 22412:2017. The encapsulation efficiency was measured via RiboGreen RNA assay (Invitrogen Quant-it RiboGreen RNA Assay Kit, protocol as described in Current Protocols 3, no. 9 (2023): e898), The results of size, polydispersity, and encapsulation efficiency are shown in Table 1 below.

[0147] Table 1. Characteristics of mRNA -LNPs of Preparation Examples

[0148] Testing Example 2: Transfection efficiency of HEK 293 cells

[0149] In this example, human embryonic kidney 293 (HEK 293) cells were seeded in a 96- well culture plate with a concentration of 5000 cells / well. After the cells were attached, the cells were washed and the m RNA -LNPs obtained in PE I and CPE I suspended in 20 mM Tris-Ci (pH 7.4) with a concentration of 20 ng mRNA per well were added separately and incubated for 24 hours. After that, the luminescent intensity (24-HR-F-Luc luminescent intensity) of each well was analyzed by a microplate reader (Agilent BioTek Synergy H 1 ). The higher luminescent intensity indicates a higher transfection efficiency.

[0150] Regarding the mRNA -LNPs of Preparation Example 1 (PHI) and Comparative Preparation Example 1 (CPE1), the size and polydispersity of the mRNA -LNPs of PEI and CPE1 are shown in Table 1 and FIGs. 14 and 15; and the transfection efficiency of the mRNA “ LNPs of PE I and CPE I is shown in FIG. 16. The mRNA -LNPs of PEI have a notably smaller size and lower polydispersity compared to the mRNA -LNPs of CPE 1 , and the transfection efficiency of the mRNA -LNPs of PEI is significantly higher than that of the mRNA -LNPs of CPE I. This comparison demonstrates the utility of the present invention at manufacturing high quality particles compared to the most commonly used manual pipette mixing.

[0151] Regarding the mRNA -LNPs of Preparation Example 2 (PE2) and Preparation Exampie 3 (PE3), the size and polydispersity of the mRNA -LNPs of PE2 and PE3 are shown in Table 1 and FIG. 17; and the transfection efficiency of the mRNA -LNPs of PE2 and PE3 are shown in FIG. 18. The mRNA -LNPs of PE3 exhibit a smaller size and lower polydispersity, and a higher transfection efficiency. This example demonstrates the utility of employing volume ratio to control particle size and polydispersity.

[0152] Regarding the mRNA -LNPs of Preparation Example 3 (PE3) to Preparation Example 5 (PE5), the size and poly dispersity of the mRNA -LNPs of PE3 to PE5 are shown in Table 1 and FIGs. 19 and 20. The mRN A -LNPs of PE3 to PE5 all exhibit small particle size and low polydispersity, which are desirable particle attributes for effective drug delivery. This example demonstrates the use of different ionizable lipids to form different lipid nanoparticle compositions to effectively encapsulate nucleic acid for potent drug delivery.

[0153] Even though numerous characteristics and advantages of the present invention have been set forth in the foregoing description, together with details of the structure and feat ures of the invention, the disclosure is illustrative only. Changes may be made in the details, especially in matters of shape, size, and arrangement of parts within the principles of the invention to the full extent indicated by the broad general meaning of the terms in which the appended claims are expressed.

Claims

WHAT IS CLAIMED IS:

1. A method for manufacturing particles, comprising the following steps:(1) providing a device for manufacturing particles, and the device comprising: a fust port for receiving a first solution; a first channel comprising a first inlet in fluid communication with the first port; and a first outlet; a second port for receiving a second solution; a second channel comprising a second inlet in fluid communication with the second port; and a second outlet; a third channel comprising a third inlet in fluid communication with the first outlet and the second outlet; a third outlet; and a terminal mixing section between the third inlet and the third outlet and configured to mix fluid flow in die third channel to manufacture particles: and a first intersection of the first outlet, the second outlet and the third inlet;(2) introducing a first solution into the first port, and a second solution into the second port;(3) applying a relative centrifugal force (RCF) of 50 xg to 32000 xg to the device to initiate fluid flow in the device and manufacture particles.

2. The method as claimed in claim 1, comprising fluid flow and mixing on a rotating frame,3. The method as claimed in claim 1 or 2fwherein the device is oriented in the centrifuge such that a principal flow direction is parallel to or angled within 90 degrees relative to a centrifugal force direction.

4. The method as claimed in any one of claims 1 to 3, wherein the first solation and second solution converge at substantially the same time.

5. The method as claimed in any one of claims 1 to 4, wherein the fluid flow and mixing are achieved only under a centrifugal force without use of an external pressure source.

6. The method as claimed in any one of claims I to 5, wherein the device is centrifuged for 1 second to 60 minutes.

7. The method as claimed in any one of claims 1 to 6, wherein a volume ratio of the first solution and the second solution is between 2:1 and 10: 1.

8. The method as claimed in any one of claims 1 to 7, wherein the particles are nanoparticles or microparticles.

9. The method as claimed in claim 8, wherein the particles are lipid nanoparticles or polymer nanoparticles.

10. The method as claimed in claim 8 or 9, wherein the nanoparticles have a diameter of about 200 nm or less, as measured by dynamic light scattering.11 . The method as claimed in claim 8, wherein the particles are lipid microparticles or polymer microparticles.

12. The method as claimed in claim 8 or 11, wherein the microparticles have a diameter of about 1000 nm or less, as measured by dynamic light scattering.

13. The method as claimed in any one of claims I to 12, wherein the particles have an encapsulation efficiency of about 60% or more.

14. The method as claimed in any one of claims 1 to 13, wherein the particles have a polydispersity index of about 0.25 or less, as measured by dynamic light scattering.

15. The method as claimed in any one of claims 1. to 14, wherein the particles are spherical or non-spherical.

16. The method as claimed in any one of claims 1 to 15, wherein the first solution comprises nucleic acids, and the second solution comprises lipids or lipid particles.

17. The method as claimed in any one of claims 1 to 15, wherein the first solution comprises aqueous buffer.

18. The method as claimed in any one of claims 1 to 15. wherein the second solution comprises ethanol.

19. The method as claimed in any one of claims 1 to 15, wherein the first solution comprises RNA.

20. The method as claimed in any one of claims 1 to 15, wherein the first solution comprises DNA.

21. The method as claimed in any one of claims 1 to 20, wherein twoor more said devices are centrifuged simultaneously to manufacture one or more distinct particles.

22. The method as claimed in any one of claims 1 to 20, wherein 24 to 96 said devices are centrifuged simultaneously to manufacture one or more distinct particles.

23. A particle made by the method of any one of claims 1 to 22.

24. A device for manufacturing particles, and the device comprising: a first port for receiving a first solution; a first channel comprising a first inlet in fluid communication with the first port; and a first outlet; a second port for receiving a second solution; a second channel comprising a second inlet in fluid communication with the second port; and a second outlet; a third channel comprising a third inlet in fluid communication with the first outlet and the second outlet: a third outlet: and a terminal mixing section between the third inlet and the third outlet and configured to mix fluid flow in the third channel to manufacture particles; and a first intersection of the first outlet, the second outlet and the third inlet;wherein the first solution in the first port flows through the first channel and the second solution in the second port flows through the second channel when a relative centrifugal force (RCF) of 50xg to 32000xg is applied to the device.

25. The device as claimed in claim 24, wherein the channels are configured so that a principal flow' direction is parallel to or angled within 90 degrees relative to a centrifugal force direction.

26. The device as claimed in claim 24 or 25, further comprising: at least one additional port, and each of the at least one additional port for receiving an additional solution; at least one additional channel, and each of the at least one additional channel comprising: an additional inlet in fluid communication with a corresponding one of the at least one additional port; and an additional outlet in fluid communication with the third channel; wherein the additional solution in each of the at least one additional port flows through the at least one additional channel when a RCF of 50xg to 32000xg is applied to the device.

27. The device as claimed in claim 26, wherein the additional outlet of one or more of the a t least one additional channel is connected to the first intersection,28. The device as claimed in claim 26 or 27, wherein the third channel further includes at least one middle mixing sectionbetween the third inlet and the terminal mixing section; and the additional outlet of each of the at least one additional channel is connected to an intersection defined on the third channel and disposed between the terminal mixing section and one of the at least one middle mixing section or between two adjacent middle mixing sections when the third channel includes two or more middle mixing sections.

29. A kit for manufacturing particles, comprising the device as claimed in any one of claims 24 to 28.

30. The kit as claimed in claim 29, further comprising an outer container.

31. The kit as claimed in claim 30, wherein the outer container collects particles from the device.

32. The kit as claimed in claim 30 or 31, wherein the outer container is a centrifuge tube of about 0.2 mL or more volume capacity.

33. The kit as claimed in claim 30 or 31, wherein the outer container is a well plate comprising 6 wells, 12 wells, 24 wells, 48 wells, or 96 wells.

34. A system lor manufacturing particles, comprising the device as claimed in any one of claims 24 to 28 and a centrifuge.

35. A system for manufacturing particles, comprising the kit as claimed in any one of claims 29 to 33 and a centrifuge.

36. The system as claimed in claim 34 or 35, wherein the centrifuge comprises a fixed-angle rotor or a swing-out rotor.