Automated apparatus for producing nanoparticles

Abstract

The present invention relates to an automated apparatus for producing nanoparticles, and a nanoparticle production method using same. Specifically, the present invention relates to the automated apparatus for producing nanoparticles, comprising: a microfluidic device; a mounting part for fastening the microfluidic device to the automated apparatus; an inlet part that supplies a fluid containing raw materials; a pneumatic pressure control part that provides pressure for fluid movement; and a recovery part that recovers the produced nanoparticles according to the condition thereof.

Description

Nanoparticle manufacturing automation device

[0001] The present invention relates to an automated device for manufacturing nanoparticles, including lipid nanoparticles (LNPs). Specifically, the present invention relates to an automated device for manufacturing nanoparticles, comprising a microfluidic device, a mounting portion for attaching the microfluidic device to the automated device, an inlet portion for supplying a fluid containing a raw material, a pneumatic control portion for providing pressure for fluid movement, and a recovery portion for recovering the manufactured nanoparticles according to conditions.

[0002] Many nanoparticle-based pharmaceuticals are being developed for the targeted delivery of therapeutics and imaging agents for the treatment and diagnosis of major diseases, including cancer, cardiovascular disease, diabetes, and Alzheimer's disease. Effective drug delivery systems can improve the absorption of poorly soluble and unstable drugs, while enhancing their therapeutic efficacy and reducing their toxic effects. This, in turn, leads to the discovery and development of effective drugs that can improve patient prognosis and quality of life.

[0003] Nanoparticles developed as pharmaceuticals are typically based on phospholipid-based lipid nanoparticles. In the early stages of technology development, emulsion or extruder methods were commonly used. However, these methods had low yields and produced non-uniform particle sizes, limiting their industrial value. Recently, microfluidic manufacturing methods have emerged as a preferred alternative to traditional lipid nanoparticle synthesis methods. This method achieves micrometer-scale microstructures, maximizing the interface between the hydrophobic fluid containing phospholipids and the hydrophilic fluid containing the pharmaceutical ingredient, enabling more efficient nanoparticle production.

[0004] Currently, the most commonly used microstructured mixers are T (or Y) mixers and ring mixers. However, manufacturing nanoparticles using microstructures requires a high level of automation for mass production. To this end, equipment for automated nanoparticle production using microfluidic devices has recently been developed. However, the most crucial aspect of automated nanoparticle manufacturing using microfluidic devices is the ability to mass-produce small, uniform nanoparticles.

[0005] Accordingly, the inventor of the present invention has continued research to achieve the above-mentioned purpose, and as a result, has recognized that a precisely calculated constant flow rate must be provided, and has completed the present invention by introducing a pressure control method to an automated device for manufacturing nanoparticles.

[0006]

[0007] [Patent Document]

[0008] (Patent Document 1) Korean Patent No. 10-2492420

[0009] [Non-patent literature]

[0010] (Non-patent Document 1) Velencia, P. et al. Single-Step Assembly of Homogenous Lipid-Polymeric and Lipid-Quantum Dot Nanoparticles Enabled by Microfluidic Rapid Mixing. ACS Nano 4, 3, 1671-1679 (2010)

[0011] (Non-patent Document 2) Rhee, M. et al. Drop Mixing in a Microchannel for Lab-on-a-Chip Platforms.Langmuir, 24 (2), 590-601 (2008)

[0012] (Non-patent Document 3) Rohit, K et al. Microfluidic Platform for Controlled Synthesis of Polymeric Nanoparticles.Nano Letters, 8, 9, 2906-12, (2008).

[0013]

[0014] The present invention aims to mass-produce uniform nanoparticles using a microfluidic device designed to maximize mixing of different fluids.

[0015] Specifically, the present invention aims to provide an automated device for manufacturing nanoparticles, which can precisely control the flow rate of a fluid using a raw material injection device controlled by a high-precision pneumatic control device and automatically classify each nanoparticle synthesized under various conditions using a rotary sample recovery device.

[0016] The present invention provides a nanoparticle manufacturing device including a microfluidic device, an inlet portion, a recovery portion, and a pneumatic control portion.

[0017] The nanoparticle manufacturing device of the present invention may additionally include a microfluidic device mounting portion.

[0018] The nanoparticle manufacturing device of the present invention may additionally include a cabinet.

[0019] The nanoparticle manufacturing device of the present invention may additionally include a power supply unit.

[0020] The microfluidic device mounting portion of the present invention may include at least one selected from the group consisting of a mounting portion cover, a microfluidic device fastening holder, and an injection valve. Here, the injection valve can control the injection of fluid delivered from the inlet portion.

[0021] The inlet of the present invention may include at least one selected from the group consisting of a tube mounting portion, a raw material injection tube, a sealing electric cylinder, a sealing gasket, an electric cylinder for opening / closing the tube mounting portion, and a pneumatic fitting.

[0022] Here, the raw material injection tube can be sealed from external gas by a sealing electric cylinder and a sealing gasket. Furthermore, the fluid pressure controlled by the pneumatic control unit can be transmitted to the microfluidic device through the pneumatic fitting and the raw material injection tube. Furthermore, the tube mounting portion can be moved forward and backward by the tube mounting portion opening and closing electric cylinder, thereby opening and closing the door.

[0023] The recovery unit of the present invention may include at least one selected from the group consisting of a tube mounting unit, a rotation motor, a recovery valve, and an electric cylinder for opening and closing the tube mounting unit.

[0024] Here, the tube mounting portion can be moved forward and backward by an electric cylinder for opening and closing the tube mounting portion, thereby opening and closing the door. In addition, the tube mounting portion can be rotated by a rotation motor, and nanoparticles synthesized in the microfluidic device can be stored in a separate recovery tube.

[0025] The pneumatic control unit of the present invention may include at least one selected from the group consisting of a primary control unit and a secondary control unit.

[0026] In the nanoparticle manufacturing device of the present invention, a microfluidic device can be fastened to a holder for fastening a microfluidic device.

[0027] In the nanoparticle manufacturing device of the present invention, the inlet and recovery portions may have an openable door structure.

[0028] In the nanoparticle manufacturing device of the present invention, a gas having a constant pressure can be supplied to the inlet through the first control unit and the second control unit. At this time, the gas can be supplied from outside.

[0029] In the nanoparticle manufacturing device of the present invention, the microfluidic device may include one or more microchannels. The height and width of the microchannels may be in the range of 1:1 to 1:5, and preferably in the range of 1:1 to 1:3.

[0030] In the above microchannel, the Reynolds number may be 10 to 500, and preferably 20 to 300. Specifically, when the height and width of the microchannel are 1:1 (X1), the Reynolds number may be 20 or more, and preferably 20 to 300. In addition, when the height and width of the microchannel are 1:3 (X3), the Reynolds number may be 150 or more, and preferably 150 to 300.

[0031] The size of the nanoparticles manufactured by the nanoparticle manufacturing device of the present invention may be 10 to 100 nm, preferably 30 to 90 nm, and more preferably 50 to 80 nm.

[0032] The polydispersity index of nanoparticles manufactured by the nanoparticle manufacturing device of the present invention may be 0.05 to 0.15, and preferably 0.10 to 0.15.

[0033] The encapsulation rate of nanoparticles manufactured by the nanoparticle manufacturing device of the present invention may be 70% or more, preferably 80% or more, and more preferably 90% or more.

[0034] In the nanoparticle manufacturing device of the present invention, a method for manufacturing nanoparticles is provided, comprising the steps of introducing a fluid from an inlet to a microfluidic device at a constant flow rate, and recovering a product generated in the microfluidic device to a recovery unit.

[0035] Here, when fluid is introduced into the microfluidic device from the inlet, the flow rate of the fluid can be controlled by the pneumatic control unit. In addition, each of the products generated in the microfluidic device can be separated and recovered by the recovery unit.

[0036] The above nanoparticle manufacturing device can be installed inside the cabinet.

[0037] The automated device for manufacturing nanoparticles of the present invention enables the uniform synthesis of nanoparticles of a desired size by supplying a fluid containing a raw material at a precisely controlled flow rate to a microfluidic device designed for the purpose of generating micro-vortices.

[0038] Additionally, the rotating rotor can automatically replace the recovery tubes to separate nanoparticles under various conditions. This allows for the identification of the optimal combination of precursors for developing the desired nanoparticles.

[0039] Therefore, the device of the present invention can efficiently perform development and mass production of nanoparticles for various purposes, minimize the cost and time for development, and enable mass production.

[0040] Figure 1 is a perspective view of a nanoparticle manufacturing device.

[0041] Figure 2 is a perspective view of the inlet and return doors of the nanoparticle manufacturing device with the doors open.

[0042] Figure 3 is an internal perspective view of a nanoparticle manufacturing device excluding the cabinet.

[0043] Figure 4 is a perspective view of the microfluidic device mounting portion with the mounting portion cover opened.

[0044] Figure 5 is a bottom perspective view of the microfluidic device mounting portion.

[0045] Figure 6 is a front view of the inlet.

[0046] Figure 7 is a perspective view of the inlet.

[0047] Figure 8 is a front view and a side view of the recovery unit.

[0048] Figure 9 is a plan view of the recovery section.

[0049] Figure 10 is a perspective view of the recovery unit.

[0050] Figure 11 is a graph comparing the sizes of LNPs when the lipid mixture concentrations are 0.66, 6.6, 13, and 20 mg / mL.

[0051] Figure 12 is a graph comparing the polydispersity index (PDI) of LNPs at lipid mixture concentrations of 0.66, 6.6, 13, and 20 mg / mL.

[0052] Figure 13 is a graph comparing the encapsulation efficiency (EE) of LNPs when the lipid mixture concentrations are 0.66, 6.6, 13, and 20 mg / mL.

[0053] Figure 14 shows the DLS results for the size of LNPs in numbers (percentages) according to lipid mixture concentrations of 0.66, 6.6, 13, and 20 mg / mL.

[0054] Figure 15 shows the DLS results for the size of LNPs in terms of volume (percentage) according to lipid mixture concentrations of 0.66, 6.6, 13, and 20 mg / mL.

[0055] Figure 16 shows the DLS results for the size of LNPs according to lipid mixture concentrations of 0.66, 6.6, 13, and 20 mg / mL, expressed as intensity (percentage).

[0056]

[0057] Hereinafter, with reference to the attached drawings, embodiments and examples of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. However, the present invention may be implemented in various forms and is not limited to the embodiments and examples described herein.

[0058] Throughout this specification, whenever a part is said to "include" a component, this means that it may include other components, but not to the exclusion of other components, unless otherwise stated.

[0059] The present invention relates to a nanoparticle manufacturing device including a microfluidic device, an inlet portion, a recovery portion, and a pneumatic control portion.

[0060] The present invention relates to a method for manufacturing nanoparticles, comprising the step of introducing a fluid containing a raw material into a microfluidic device at a constant flow rate through pneumatic pressure from an inlet portion in the nanoparticle manufacturing device of the present invention, and the step of classifying and recovering a product in a recovery portion.

[0061] The term "microfluidic device" as used herein means a device including microchannels, etc., that allow fluid to flow on a substrate made of various materials including plastic, glass, metal or silicon containing organic polymer materials.

[0062] The term "microchannel" as used herein means a microscopic channel having dimensions of millimeters, micrometers, or nanometers through which a fluid can flow, and is also referred to as a "mixed channel" in the present invention.

[0063] The microfluidic device used in the present invention may be the device described in Korean Patent No. 10-2631907.

[0064] The term "resistance" used herein refers to the resistance in Pascal·second / meter cubic (Pa·s / m) when a fluid containing a raw material to be injected during nanoparticle manufacturing is injected at a constant flow rate into a microchannel of a microfluidic device. 3 ) refers to the pressure per unit volume received by a fluid in proportion to the time having the dimension of .

[0065] The term "pressure" or "pneumatic pressure" as used herein refers to the pressure controlled during the production of nanoparticles, which has dimensions of Pascal (Pa), kiloPascal (kPa), millibars (mbar), or bars, and means the power that enables the injection of fluid and the production of nanoparticles through precise control.

[0066] Hereinafter, the nanoparticle manufacturing device of the present invention will be described in detail with reference to the drawings. The present invention may include a nanoparticle manufacturing device embodied in the form of FIGS. 1 to 10, but is not limited thereto, and includes various modifications that can be implemented by a person skilled in the art to which the present invention pertains.

[0067] The nanoparticle manufacturing device (1) of the present invention includes a microfluidic device mounting portion (20), an inlet portion (30), a recovery portion (40), and a pneumatic control portion (50).

[0068] Referring to FIG. 1, the nanoparticle manufacturing device (1) of the present invention can fasten a microfluidic device (201) to a holder (22) for fastening a microfluidic device.

[0069] Referring to FIG. 2, the inlet (30) and the recovery (40) have an open / close door structure, and each may include a raw material tube mounting portion (31) capable of fastening a tube and a rotary recovery tube mounting portion (41).

[0070] Referring to FIG. 3, the nanoparticle manufacturing device (1) may include a pneumatic control unit (50) and may deliver a gas of a constant pressure to the inlet unit (30) through the primary control unit (51) and the secondary control unit (52). At this time, the gas supplied to the pneumatic control unit (50) may be supplied from outside the device.

[0071] The gas supply source supplied to the nanoparticle manufacturing device (1) may be a gas storage tank.

[0072] Referring to FIG. 5, the microfluidic device mounting portion (20) may include an injection valve (23) for controlling the injection of a fluid containing a raw material delivered from the inlet portion (30).

[0073] Referring to Fig. 6, the raw material tube connected to the raw material tube mounting portion (31) can be sealed from the outside air by a sealing electric cylinder (33) and a sealing gasket (34), and a pneumatic fitting (36) connected to a pneumatic secondary control device (52) and a raw material injection tube (32) can be used to apply a constant pressure to the raw material fluid inside the tube and deliver it to the microfluidic device (201). In the present invention, the raw material injection tube can include a double-sided injection tube and a central injection tube.

[0074] Referring to Fig. 7, the raw material tube mounting part (31) can be moved forward / backward by the raw material tube mounting part opening / closing electric cylinder (33) to open and close the door.

[0075] Referring to FIGS. 8 and 10, the rotary recovery tube mounting portion (41) rotates by a rotation motor (42), thereby allowing nanoparticles synthesized under different conditions to be stored in different recovery tubes.

[0076] Referring to Fig. 9, the rotary recovery tube mounting part (41) can be moved forward / backward by the recovery tube mounting part opening / closing electric cylinder (43) to open and close the door.

[0077]

[0078] [Example 1]

[0079] Resistance range according to the mixing channel size and number of channel units in the microfluidic device

[0080]

[0081] The resistance at each channel inlet (both inlets and the central inlet) applied according to the mixing channel size and the number of channel units of the microfluidic device of the present invention is shown in Table 1 below. Even if the mixing channel size is the same, the resistance may vary depending on the number of units, and the dynamic viscosity of all channel parts was based on 20°C.

[0082]

[0083] Resistance value according to the size of the microfluidic device and the number of channel units Number of channel units X 1 (single unit) X 1 (6-unit) X 3 (single unit) X 3 (18-unit) Microchannel size H (mm) 0.2 0.2 0.2 0.2 0.2 W (mm) 0.2 0.2 0.6 0.6 L (mm) 8 10.9 6 5.7 Resistance (Pa s / m) 3 )Aqueous phase(Side inlets)8.1Х10 10 1.8Х10 10 9.5Х10 9 5.3Х10 8 Organic phase(Center inlet)1.6Х10 11 3.7Х10 10 1.9Х10 10 1.1Х10 9

[0084] H: height of microchannel

[0085] W: width of the microchannel

[0086] L: Length of microchannel

[0087]

[0088] [Example 2]

[0089] Pressure range for nanoparticle synthesis depending on the type of microfluidic device

[0090]

[0091] The flow rate and pressure according to the Reynolds number required for driving the microfluidic device of the present invention are shown in Tables 2 to 9 below, depending on the type of each microfluidic device.

[0092] In particular, Tables 3 and 4 show the pressure, flow rate, and Reynolds number applied to each microchannel of a parallel microfluidic device consisting of 6 units, and Tables 8 and 9 show the pressure, flow rate, and Reynolds number applied to each microchannel of a parallel microfluidic device consisting of 18 units.

[0093]

[0094] Pressure and Reynolds number according to the required flow rate of both side inlets of the microfluidic device of X1 (single unit) X1 (single unit) Aqueous phase (side inlets) Pressure (Pa) Flow rate [both side inlets] Re10 -9 m 3 / smL / min7609.30.56101500191.1203000372.2406000734.480120001508.816023000280173003800047028500

[0095]

[0096] Pressure and Reynolds number according to the required flow rate at the center channel inlet of the microfluidic device of X1 (single unit)X1 (single unit)Organic phase (center inlet)Pressure (Pa)Flow rateRe10 -9 m 3 / smL / min2801.70.10105503.40.202011006.70.40402200130.80804300271.61608300513.130014000855.1500

[0097]

[0098] Pressure and Reynolds number according to the required flow rate of both side inlets of the microfluidic device of X1 (6-unit) X1 (6-unit) Aqueous phase (side inlets) Pressure (Pa) Flow rate [Total of both side inlets] Re10 -9 m 3 / smL / min1000563.41020001106.72040002201340810044026801600088053160310001700100300520002800170500

[0099]

[0100] Pressure and Reynolds number according to the required flow rate at the center channel inlet of a microfluidic device of size X1 (6-unit)X1 (6-unit)Organic phase (center inlet)Pressure (Pa)Flow rateRe10 -9 m 3 / smL / min380100.6110750201.2201500402.4403000804.88059001609.616012000310183001900051031500

[0101]

[0102] Pressure and Reynolds number according to the required flow rate of both side inlets of the microfluidic device of X3 (single unit) X3 (single unit) Aqueous phase (side inlets) Pressure (Pa) Flow rate [both side inlets] Re10 -9 m 3 / smL / min25002601610037003902415050005303220075007904730013000130079500

[0103]

[0104] Pressure and Reynolds number according to the required flow rate of the center channel inlet of the microfluidic device of X3 (single unit)X3 (single unit)Organic phase (center inlet)Pressure (Pa)Flow rateRe10 -9 m 3 / smL / min900472.91001400744.31501800955.720028001458.6300460024014500

[0105]

[0106] Pressure and Reynolds number according to flow rate applied to all 36 side inlets of the X3 (18-unit) microfluidic device X3 (18-unit) Aqueous phase (side inlets) Pressure (Pa) Flow rate [Total 36 side inlets] Re10 -9 m 3 / smL / min25004700280100380071004301505000940057020076001400085030013000240001400500

[0107]

[0108] Pressure and Reynolds number according to flow rate applied to all 18 center channel inlets of the X3 (18-unit) microfluidic device X3 (18-unit) Organic phase (center inlet) Pressure (Pa) Flow rate [18 total] Re10 -9 m 3 / smL / min900850521001400130077150180017001002002800260016030046004300260500

[0109]

[0110] The pressures shown in Tables 2 to 9 above were calculated using the equation below according to the resistance of each microfluidic channel shown in Table 1.

[0111]

[0112] In the above equation, Q represents flow velocity, P represents pressure, and R represents resistance.

[0113] Therefore, as shown in Tables 2 to 9 above, since the resistance is different depending on the size of the mixing channel and the number of channel units of the microfluidic device of the present invention, it can be seen that different pressures are applied depending on the type of microfluidic device.

[0114] Additionally, depending on the type of each channel (double-channel inlet and central channel inlet), the pressure must be controlled separately. For the X1 (single unit) microfluidic device, the pressure to be controlled at the double-channel inlet is 760 Pa to 38000 Pa, and the pressure to be controlled at the central channel inlet is 280 Pa to 14000 Pa.

[0115] For the microfluidic device of X1 (6-unit), the pressure to be controlled at the inlets of both channels is 1000 Pa to 52000 Pa, and the pressure to be controlled at the inlet of the central channel is 380 Pa to 19000 Pa.

[0116] For the microfluidic device of X3 (single unit), the pressure to be controlled at the inlets of both channels is 2500 Pa to 13000 Pa, and the pressure to be controlled at the inlet of the central channel is 900 Pa to 4600 Pa.

[0117] For the X3 (18-unit) microfluidic device, the pressure to be controlled at the inlets of both channels is 2500 Pa to 13000 Pa, and the pressure to be controlled at the inlet of the central channel is 900 Pa to 4600 Pa.

[0118]

[0119] [Example 3]

[0120] Optimization of the operating pressure range for nanoparticle synthesis according to the type of microfluidic device.

[0121]

[0122] As shown in Tables 2 to 9 above, the Reynolds number in the microfluidic device X1 is 10 to 500, and the Reynolds number in the microfluidic device X3 is 100 to 500. However, in order to ensure a mixing efficiency of 90% or more for uniform synthesis of nanoparticles, the Reynolds number in the microfluidic device X1 must be 20 or more, and the Reynolds number in the microfluidic device X3 must be 150 or more.

[0123] In addition, when using biomolecules as raw materials for manufacturing nanoparticles, the shear rate should be 10 to minimize damage to the biomolecules. 5 s -1 It is desirable to manufacture within the following range.

[0124] Table 10 below shows the shear rate according to each Reynolds number, and the shear rate is 10 when the Reynolds number is 300 or less. 5 s -1 You can see that below.

[0125]

[0126] Shear rate as a function of Reynolds number (10 5 s -1 )100.03200.07400.1800.31600.53001.05002.0

[0127] Therefore, when using the microfluidic device of X1 (single unit), the pressure at which nanoparticles can be synthesized is 1500 to 38000 Pa for the double channel inlets and 550 to 14000 Pa for the central channel inlet. When using the microfluidic device of X1 (6-unit), the pressure at which nanoparticles can be synthesized is 2000 to 52000 Pa for the double channel inlets and 750 to 19000 Pa for the central channel inlet. In addition, when using the microfluidic device of X1 (single unit), the pressure at which nanoparticles can be synthesized while minimizing damage to biomaterials is 1500 to 23000 Pa for the double channel inlets and 550 to 8300 Pa for the central channel inlet. Using the X1 (6-unit) microfluidic device, the pressure at which nanoparticles can be synthesized while minimizing damage to biomaterials is 2000 to 31000 Pa for the double-channel inlet and 750 to 12000 Pa for the central channel inlet.

[0128] When using a microfluidic device of X3 (single unit), the pressure at which nanoparticles can be synthesized is 3700 to 13000 Pa for the double channel inlets and 1400 to 4600 Pa for the central channel inlet. When using a microfluidic device of X3 (18-unit), the pressure at which nanoparticles can be synthesized is 3800 to 13000 Pa for the double channel inlets and 1400 to 4600 Pa for the central channel inlet.

[0129] When using a microfluidic device of X3 (single unit), the pressure at which nanoparticles can be synthesized while minimizing damage to biomaterials is 3700 to 7500 Pa for the double channel inlets and 1400 to 2800 Pa for the central channel inlet. When using a microfluidic device of X3 (18-unit), the pressure at which nanoparticles can be synthesized while minimizing damage to biomaterials is 3800 to 7600 Pa for the double channel inlets and 1400 to 2800 Pa for the central channel inlet.

[0130]

[0131] [Example 4]

[0132] Production of nanoparticles according to lipid mixture concentration using a microfluidic device

[0133]

[0134] Nanoparticles containing phospholipids (DSPC) and mRNA were manufactured using the microfluidic device of the present invention by the following method.

[0135] A solution of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) in absolute ethanol and an mRNA solution in citrate buffer were prepared. Then, approximately 1 mL of the DSPC solution in absolute ethanol at a concentration of 0.66 to 20 mg / mL was placed in a 50 mL conical tube, and approximately 3 mL of the mRNA solution in citrate buffer at a concentration of 0.0042 to 0.13 mg / mL was placed in another 50 mL conical tube.

[0136] In order to synthesize in the microfluidic device of the present invention, a 50 mL conical tube with a 50 mL conical tube cap was mounted on the precursor rack, and after flowing ethanol to wash the microfluidic device of the present invention, ethanol and DW were additionally flowed to minimize air bubbles in the microfluidic device. Thereafter, using the microfluidic device of the present invention, the injection flow rate of the DSPC solution was set to 0.8 mL / min, and the injection flow rate of the mRNA solution was set to 4.4 mL / min. The manufactured nanoparticles were obtained through the outlet of the microfluidic device of the present invention, and the obtained nanoparticles were placed in a dialysis cassette and dialyzed for 16 hours at refrigeration or room temperature.

[0137] The size distribution change of the manufactured nanoparticles according to the aggregation phenomenon of the nanoparticles was measured using the dynamic light scattering method (DLS) using Zetasizer Nano ZS, and the results are shown in Table 11, Figures 11 and 12 below.

[0138] As shown in Table 11, Figures 11 and 12, the sizes of LNPs at lipid mixture concentrations of 0.66, 6.6, 13 and 20 mg / mL were measured to be 55, 70, 72 and 72 nm, respectively, and the PDI at this time was 0.124, 0.109, 0.111 and 0.143, respectively, indicating that they were synthesized very uniformly.

[0139] Lipid mixture concentration (mM)Lipid mixture concentration (mg / mL)Average LNP size (nm)PDI encapsulation rate (%)1.10.66550.12493116.6700.109992113720.111983220720.14394

[0140] The nucleic acid encapsulation efficiency (EE) was calculated using the values ​​obtained by detecting total mRNA and free mRNA based on the fluorescence intensity in each mRNA LNP using the following formula ( ) was calculated according to the following.

[0141] As shown in Table 11 and Figure 13, the nucleic acid encapsulation rates of each LNP according to the lipid mixture concentrations of 0.66, 6.6, 13, and 20 mg / mL were confirmed to be 93, 99, 98, and 94%, respectively.

[0142] It was confirmed that nanoparticles of a desired concentration can be uniformly synthesized using the microfluidic device of the present invention.

[0143]

[0144] [Explanation of symbols]

[0145] 1: Nanoparticle manufacturing device

[0146] 10: Cabinet

[0147] 11: Cabinet top

[0148] 20: Microfluidic device mounting section

[0149] 201: Microfluidic Devices

[0150] 21: Mounting cover

[0151] 22: Holder for attaching microfluidic devices

[0152] 23: Injection valve

[0153] 30: Inlet

[0154] 31: Tube mounting section for raw materials

[0155] 32: Tube for raw material injection

[0156] 33: Tube sealing electric cylinder for raw materials

[0157] 34: Sealing gasket

[0158] 35: Electric cylinder for opening / closing the tube mounting part for raw materials

[0159] 351: Electric cylinder motor

[0160] 36: Pneumatic fittings

[0161] 40: Recovery Department

[0162] 41: Rotating recovery tube mounting part

[0163] 42: Rotation motor

[0164] 43: Electric cylinder for opening / closing the recovery tube mounting part

[0165] 431: Electric cylinder motor

[0166] 50: Pneumatic control unit

[0167] 51: Pneumatic primary control device

[0168] 52: Pneumatic secondary control device

[0169] 60: Power supply

Claims

1. A nanoparticle manufacturing device comprising a microfluidic device, an inlet section, a recovery section, and a pneumatic control section.

2. A nanoparticle manufacturing device according to claim 1, further comprising a microfluidic device mounting portion.

3. A nanoparticle manufacturing device according to claim 1, further comprising a cabinet.

4. A nanoparticle manufacturing device according to claim 2, wherein the microfluidic device mounting portion includes at least one selected from the group consisting of a mounting portion cover, a microfluidic device fastening holder, and an injection valve.

5. A nanoparticle manufacturing device according to claim 1, wherein the inlet includes at least one selected from the group consisting of a tube mounting portion, a raw material injection tube, a sealing electric cylinder, a sealing gasket, an electric cylinder for opening / closing the tube mounting portion, and a pneumatic fitting.

6. A nanoparticle manufacturing device according to claim 1, wherein the recovery unit includes at least one selected from the group consisting of a tube mounting unit, a rotation motor, a recovery valve, and an electric cylinder for opening / closing the tube mounting unit.

7. A nanoparticle manufacturing device according to claim 1, wherein the pneumatic control unit includes at least one selected from the group consisting of a primary control unit and a secondary control unit.

8. A nanoparticle manufacturing device according to claim 4, wherein the microfluidic device is fastened to a holder for fastening the microfluidic device.

9. A nanoparticle manufacturing device in the first paragraph, wherein the inlet and the return have an openable door structure.

10. A nanoparticle manufacturing device according to claim 7, wherein a gas having a constant pressure is provided to the inlet through the first control unit and the second control unit.

11. A nanoparticle manufacturing device according to claim 10, wherein the gas is supplied from outside.

12. A nanoparticle manufacturing device in the fourth paragraph, wherein the injection valve is for controlling the injection of fluid delivered from the inlet.

13. A nanoparticle manufacturing device according to claim 5, wherein the raw material injection tube can be blocked from external gas by a sealed electric cylinder and a sealing gasket.

14. A nanoparticle manufacturing device in which, in the fifth paragraph, the pressure of the fluid controlled by the pneumatic control unit is transmitted to the microfluidic device through the pneumatic fitting and the raw material injection tube.

15. A nanoparticle manufacturing device in the fifth paragraph, wherein the tube mounting portion is moved forward / backward by an electric cylinder for opening and closing the tube mounting portion, thereby opening and closing the door.

16. A nanoparticle manufacturing device in accordance with claim 6, wherein the tube mounting portion is rotated by a rotation motor, and nanoparticles synthesized in a microfluidic device can be stored in a separate recovery tube.

17. A nanoparticle manufacturing device in accordance with claim 6, wherein the tube mounting portion is moved forward / backward by an electric cylinder for opening and closing the tube mounting portion, thereby opening and closing the door.

18. A nanoparticle manufacturing device according to claim 1, wherein the microfluidic device may include one or more microchannels.

19. A nanoparticle manufacturing device according to claim 18, wherein the height and width of the microchannel have a ratio of 1:1 to 1:

5.

20. A nanoparticle manufacturing device in claim 18, wherein the Reynolds number in the microchannel is 10 to 500.

21. A nanoparticle manufacturing device in claim 18, wherein the Reynolds number in the microchannel is 20 to 300.

22. A nanoparticle manufacturing device according to claim 1, wherein the nanoparticles manufactured by the nanoparticle manufacturing device have a size of 10 to 100 nm.

23. A nanoparticle manufacturing device according to claim 1, wherein the polydispersity index of the nanoparticles manufactured by the nanoparticle manufacturing device is 0.05 to 0.

15.

24. A nanoparticle manufacturing device in accordance with claim 1, wherein the nanoparticles manufactured by the nanoparticle manufacturing device have an encapsulation rate of 90% or more.

25. In the nanoparticle manufacturing device of paragraph 1, A step of introducing a fluid into a microfluidic device from an inlet at a constant flow rate, and A method for manufacturing nanoparticles, comprising a step of recovering a product generated in a microfluidic device to a recovery unit.

26. A method for manufacturing nanoparticles, wherein, in paragraph 25, when a fluid is introduced into a microfluidic device from an inlet, the flow rate of the fluid is controlled by a pneumatic control unit.

27. A method for manufacturing nanoparticles, wherein each of the products in paragraph 25 is separated and recovered in a recovery unit.

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