Microfluidic device for nanoencapsulating mRNA in lipid nanoparticles

Abstract

The utility model relates to devices for producing nanocomponents of a controllable composition and makes it possible to carry out the encapsulation of mRNA in lipid nanoparticles, which is one of the key processes in the development and production of mRNA vaccines. The technical result is that of improving the performance of a process of nanoencapsulating mRNA in lipid nanoparticles. What is proposed is a device consisting of a microfluidic cartridge containing a microstructure of channels for allowing the nanoencapsulation of mRNA in lipid nanoparticles by microfluidic mixing, an elastic sealing element and an adapter. The channels of the cartridge are hermetically sealed by mechanically clamping the cartridge, together with the sealing element, to a flat surface of a plate of the adapter and tightening the plates of the adapter by means of screws. This technical solution makes it possible to carry out a continuous process of nanoencapsulating mRNA in lipid nanoparticles at high flow speeds and with large volumes of liquid.

Description

[0001] A microfluidic device for high-throughput nanoencapsulation of mRNA into lipid nanoparticles with tailored characteristics

[0002] The utility model relates to the field of devices for producing nanocomponents of controlled composition.

[0003] A device for mixing liquids on a chip is known, consisting of two plates hermetically sintered together with a channel cut into one of the plates. The channel has two input points, two destabilization channels for the supplied flows, a channel mixing intersection with a mixing point, a mixing channel, and a product outlet point [1]. However, this device does not allow for mixing liquids at high flow rates and does not allow for high-throughput nanoencapsulation of mRNA in lipid nanoparticles.

[0004] A device for producing lipid nanoparticles of a given size is also known [2]. This device uses a cartridge containing a microstructure with ribbed channels for mixing liquids. However, this device also does not accommodate mixing liquids at high flow rates. Furthermore, the microstructure used contains elements with characteristic dimensions smaller than 50 µm, making it difficult to manufacture the cartridge using injection molding, which is preferred for mass production.

[0005] The closest to the proposed utility model in terms of its technical essence and achievable effect is a microfluidic mixing device based on toroidal mixing elements [3]. This device utilizes a cartridge containing a microstructure consisting of a series of toroidal mixing elements, within which fluids are mixed using Dean eddy currents. This device enables nanoencapsulation of mRNA in lipid nanoparticles of controlled size with high efficiency.

[0006] The disadvantages of this device include the inability to mix liquids at high flow rates and, as a consequence, the low productivity of the process of nanoencapsulation of mRNA into lipid nanoparticles.

[0007] The technical result, which the utility model is aimed at obtaining, is an increase in the productivity of the process of nanoencapsulation of mRNA in lipid nanoparticles with specified characteristics (geometric and electrokinetic parameters, nanoencapsulation efficiency).

[0008] The technical result is achieved in a microfluidic device for the high-throughput nanoencapsulation of mRNA into lipid nanoparticles with specified characteristics, consisting of a microfluidic cartridge containing a microstructure of channels that enables nanoencapsulation of mRNA into lipid nanoparticles via microfluidic mixing, an elastic sealing element, and an adapter. The device is characterized in that the cartridge channels are sealed by mechanically pressing the cartridge and sealing element against the flat surface of an adapter plate while tightening the adapter plates with screws. The microfluidic cartridge and sealing element are made of polymeric materials with high bioinertness and chemical resistance. The adapter can be made of stainless steel. The sealing element can be attached to the cartridge using ultrasonic welding or heat sealing.

[0009] The described device design allows for mixing reagents at flow rates of at least 100 ml / min for at least 4 hours, without depressurizing the cartridge or disturbing its geometry, and without altering the specified characteristics of the nanocomponents in the final product. The reagents only come into contact with the microfluidic cartridge and sealing element, which are made of polymeric materials with high bioinertness and chemical resistance.

[0010] Fig. 1 shows a schematic view of the disassembled device.

[0011] Microfluidic cartridge 1, in which a microstructure of channels 2 is formed, providing nanoencapsulation of mRNA in lipid nanoparticles by microfluidic mixing, is positioned using pins 3 relative to the bottom plate of adapter 4 such that two inlet through-holes 5 and one outlet through-hole 6 of the microfluidic cartridge coincide with two inlet ports 7 and one outlet port 8 of the adapter, intended for connecting tubes for introducing reagents and removing the product. Sealing of the cartridge channels is achieved by mechanically pressing the cartridge with sealing element 9 against the flat surface of the top plate of adapter 10 while tightening the adapter plates with screws 11.

[0012] Fig. 2 shows the assembled device. The device synthesizes lipid nanoparticles with specified characteristics using microfluidic mixing. Reagent injection tubes are connected to the device's inlet ports, and a product output tube is connected to the outlet port. Reagents are fed into the device at specified flow rates, which are necessary to produce output nanocomponents with the desired characteristics. The cartridge's channel microstructure ensures efficient mixing of the reagents and highly effective nanoencapsulation of mRNA into lipid nanoparticles of controlled size. The device's design allows for mixing reagents at high flow rates over extended periods of time, without depressurizing the cartridge, compromising its geometry, or altering the desired characteristics of the output nanocomponents.

[0013] For example, using the device it is possible to obtain lipid nanoparticles with the following characteristics: hydrodynamic particle diameter from 80 to 90 nm, polydispersity index from 0.1 to 0.2, electrokinetic potential from -10 to -5 mV, nanoencapsulation efficiency from 80% to 90%, by mixing an aqueous solution of mRNA and an alcoholic solution of lipids at a cartridge outlet rate of 100 ml / min in a volume of 20 liters of product at the outlet. At the same time, when carrying out a similar process in a cartridge manufactured using the technology [3], after obtaining 2 liters of product, the boundaries of the specified geometric parameters are exceeded.

[0014] Thus, the technical result achieved using the claimed utility model consists in increasing the productivity of the process of nanoencapsulation of mRNA in lipid nanoparticles.

[0015] SOURCES OF INFORMATION:

[0016] 1. Russian Federation Patent No. RU2724254C 1

[0017] 2. US Patent No. US 10843194V2

[0018] 3. US Patent No. US10835878B2 - prototype.

Claims

Invention formula 1. A microfluidic device for high-performance nanoencapsulation of mRNA into lipid nanoparticles with specified characteristics, consisting of a microfluidic cartridge in which a microstructure of channels is formed that ensures nanoencapsulation of mRNA into lipid nanoparticles by microfluidic mixing, an elastic sealing element and an adapter, characterized in that the sealing of the cartridge channels is carried out by mechanically pressing the cartridge with the sealing element to the flat surface of the adapter plate when tightening the adapter plates with screws, wherein the microfluidic cartridge and the sealing element are made of polymeric materials that have high bioinertness and chemical resistance.

2. A microfluidic device according to claim 1, characterized in that the adapter is made of stainless steel.

3. The microfluidic device according to claim 1, characterized in that the sealing element is attached to the cartridge using ultrasonic welding.

4. The microfluidic device according to claim 1, characterized in that the sealing element is attached to the cartridge by means of thermal sealing. 4 SUBSTITUTE SHEET (RULE 26)

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