MIXER FOR GENERATING PARTICLES

Abstract

A mixer for generating particles, comprising a first mixing unit, wherein the first mixing unit comprises a first channel (702) and a second channel (701), the first channel (702) comprises a rectilinear channel, the second channel (701) comprises a curvilinear channel. The mixer is particularly suitable for producing nanoparticles, and the mixing efficiency can be improved. A microfluidic hybrid chip cartridge prepared by the mixer is also provided.

Claims

1. A mixer for generating particles, comprising a first mixing unit, wherein the first mixing unit comprises a first channel and a second channel, and the first channel comprises a rectilinear channel, and the second channel comprises a curvilinear channel.

2. The mixer according to claim 1, wherein the first channel comprises a first inlet and a first outlet, the second channel comprises a second inlet and a second outlet, the first inlet being in fluid communication with the second inlet, and the first outlet being in fluid communication with the second outlet.

3. The mixer according to claim 1, wherein the mixing unit further comprises a first converging region, the first converging region being in communication with the first inlet of the first channel and the second inlet of the second channel to divert a fluid.

4. The mixer according to claim 3, wherein the mixing unit further comprises a second converging region, the second converging region being in communication with the first outlet of the first channel and the second outlet of the second channel to converge fluids.

5. The mixer according to claim 1, wherein the curvilinear channel of the second channel comprises a semi-circular or arc-shaped channel.

6. The mixer according to claim 1, wherein the second channel further comprises a rectilinear initial channel, the initial channel being disposed in the upstream of the curvilinear channel.

7. The mixer according to claim 6, wherein a length of the initial segment channel is less than or equal to ⅓ of a length of the second channel.

8. The mixer according to claim 6, wherein an included angle between the initial channel and the first channel is an acute angle of less than 90 degrees.

9. The mixer according to claim 3, wherein the mixer further comprises a premixing channel, the premixing channel being in communication with the first converging region in configure to mix two different fluids.

10. The mixer according to claim 9, wherein the mixer further comprises a first transporting channel for transporting a first fluid and a second transporting channel for transporting a second fluid, the first and second transporting channels being in fluid communication with the premixing channel.

11. The mixer according to claim 2, wherein the mixer further comprises a second mixing unit comprising a third channel and a fourth channel, wherein the third channel comprises a curvilinear channel and the fourth channel comprises a rectilinear channel.

12. The mixer according to claim 11, wherein the third channel comprises a third inlet and the fourth channel comprises a fourth inlet.

13. The mixer according to claim 12, wherein the inlet of the fourth channel is adjacent to the outlet of the second channel of the first mixing unit, or the inlet of the fourth channel and the outlet of the second channel of the first mixing unit are on the same side of the channel, or the third inlet of the third channel is disposed opposite the outlet of the first channel of the first mixing unit.

14. The mixer according to claim 12, wherein the fourth channel is disposed at an obtuse angle of greater than 90 degrees with the first channel.

15. The mixer according to claim 12, wherein the third channel further comprises a rectilinear initial channel in an upstream side of the curvilinear channel, the initial channel being a partial extension of the first rectilinear channel.

16. The mixer according to claim 12, wherein the mixer comprises a third converging region, a part of a fluid in the third converging region enters into the third channel and a part of the fluid in the third converging region enters into the second channel.

17. The mixer according to claim 1, wherein the mixer further comprises a second mixing unit comprising a third channel and a fourth channel, wherein the third channel comprises a curvilinear channel and the fourth channel comprises a rectilinear channel, the third channel and the first channel are on the same side of the mixing unit, and the fourth channel and the second channel are on the other same side of the mixing unit.

18. The mixer according to claim 1, wherein the mixer further comprises a second mixing unit, wherein the first mixing unit is located upstream side of the second mixing unit, and the second mixing unit comprises a third channel and a fourth channel, wherein the third channel comprises a curvilinear channel and the fourth channel comprises a rectilinear channel; and the fourth channel is taken as a reference, the curvilinear channel of the first mixing unit and the curvilinear channel of the second mixing unit are respectively positioned on either side of the fourth channel.

19. A mixer according to any one of claims 1 to 18, wherein all channels are of the same widths or the same depths.

20. A mixer according to any one of claims 1 to 19, wherein a cross-sections of the channels are rectangular.

21. A mixer for generating a nanoparticle, comprising N mixing units, wherein each of the mixing units comprises a first channel comprising a rectilinear channel, and a second channel comprising a curvilinear channel, the first channel having a first inlet and a first outlet, the second channel having a second inlet and a second outlet, the first inlet and the second inlet being in fluid communication, wherein N is a natural integer from 1 to 6.

22. A mixer for generating a microparticle, comprising a first mixing unit, wherein the first mixing unit comprises a first channel for receiving a first fluid and a second channel for receiving a second fluid, wherein a flow path of the first fluid in the first channel is smaller than a flow path of the second fluid in the second channel.

23. A mixer for generating a microparticle, comprising a first mixing unit, wherein the first mixing unit comprises a first channel for receiving a first fluid and a second channel for receiving a second fluid, wherein a length of the first channel is less than a length of the second channel.

24. A mixer for a nanoparticle, comprising N+1 mixing units, the N.sup.th mixing unit comprising an a.sup.th rectilinear channel and an a+1.sup.th curvilinear channel, the a.sup.th rectilinear channel comprising an a.sup.th fluid inlet and an a.sup.th fluid outlet, the a+1.sup.th curvilinear channel comprising an a+1.sup.th flow inlet and an a+1.sup.th fluid outlet, wherein N is a natural integer equal to or greater than 1, and a is a natural number greater than or equal to 1.

25. The mixer according to claim 24, wherein the fluid inlet of the a.sup.th rectilinear channel and the fluid inlet of the a+1.sup.th curvilinear channel comprises an a.sup.th converging region to divert fluids at the converging region; or, the fluid outlet of the a.sup.th rectilinear channel and the fluid outlet of the a+1.sup.th curvilinear channel being in communication with an a+1.sup.th converging region to mix or converge or merge a fluid from the two channels.

26. The mixer according to claim 24, wherein the N+1.sup.th mixing unit comprises an a+2.sup.th rectilinear channel and an a+3.sup.th curvilinear channel, the a+2.sup.th rectilinear channel comprises an a+2.sup.th fluid inlet and an a+2.sup.th fluid outlet, and the a+3.sup.th curvilinear channel comprises an a+3.sup.th fluid inlet and an a+3.sup.th fluid outlet.

27. The mixer according to claim 26, wherein the a.sup.th fluid outlet is disposed opposite to the a+3.sup.th fluid inlet.

28. The mixer according to claim 26, wherein an a+1.sup.th fluid outlet is disposed adjacent to an a+2.sup.th fluid inlet or on the same side of a channel.

29. The mixer according to claim 26, wherein an upstream side of the curvilinear channel comprises a rectilinear channel comprising the fluid inlet of the curvilinear channel.

30. The mixer of any one of claims 21-29, wherein the mixer comprises a pre-premixing channel for flowing fluid into the first and second channels, the pre-premixed fluid channel being in the upstream sides of the first and second channels, or in the upstream of the rectilinear channel and the a+1.sup.th curvilinear channel, where a=1.

31. The mixer according to claim 30, wherein the pre-premixing channel comprises a mixed fluid of the first and second fluids.

32. The mixer according to claim 31, wherein the first fluid comprises a nucleic acid and the second fluid comprises a polymer.

33. The mixer according to claim 31, wherein the first fluid comprises a nucleic acid and the second fluid comprises a lipid component.

34. The mixer according to claim 31, wherein the first fluid comprises a microparticle formed from a nucleic acid and a polymer, and the second fluid comprises a lipid component.

35. A method for preparing a microparticle, the method comprising: providing the mixer according to any one of claims 1 to 34, passing a fluid from a premixing channel into a first mixing unit, wherein one part of the fluid enters a first channel of the first mixing unit and another part of the fluid enters a second channel of the first mixing unit.

36. The method according to claim 35, wherein a premixed fluid flows in through a first inlet of a first channel in communication with a first converging region and then through a second inlet of a second channel.

37. The method according to claim 30, wherein a fluid passing through the first and second channels of the first mixing unit converges at a second converging region.

38. The method according to claim 37, wherein a fluid from the first converging region enters the third and fourth channels, respectively, through inlets of the third and fourth channels of the second mixing unit in communication with the third converging region.

39. The method according to claim 36, wherein the fluid in the first mixing unit is flowed by externally applying pressure to the channel externally applying pressure to the channel.

40. The method according to claim 37, wherein the first and second fluids are premixed in a premixing channel.

41. A method for preparing a microparticle, the method comprising providing a mixed fluid, passing one part of the fluid through a first channel, and passing a rest part of the fluid through a second channel, wherein a path through which the fluid passes in the first channel is less than a path through which the fluid passes in the second channel.

42. The method according to claim 41, wherein the fluid comprises one or more of a nucleic acid, a polymer, or a lipid component substance.

43. The method according to claim 41, wherein the first channel comprises a rectilinear channel and the second channel comprises a curvilinear channel.

44. The method according to claim 41, wherein a premixing channel is provided in the upstream sides of the first and second channels, a first fluid and a second fluid being mixed into a mixed fluid in the premixing channel.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0250] FIG. 1 is a schematic diagram showing the structure of a microfluidic hybrid chip cartridge containing microfluidic chips in one Example;

[0251] FIG. 2 is a schematic diagram showing the structure of a microfluidic chip of a mixer in one Example;

[0252] FIG. 3 is a schematic perspective view of a mixing unit of microparticle channels in one Example

[0253] FIG. 4 is an enlarged structural view of a mixing unit in one Example;

[0254] FIG. 5 is a schematic view showing the structure of the mixer provided in Example 1;

[0255] FIG. 6 is a cross-sectional view A-A of FIG. 5, and FIG. 3 is a schematic view showing the flow direction of the sample in the mixer;

[0256] FIG. 7 is a schematic view showing the flow direction of the sample in the mixer provided in Example 1;

[0257] FIG. 8 is a schematic view showing the structure of the microfluidic hybrid chip provided in Example 2;

[0258] FIG. 9 is a schematic view showing the structure of the microfluidic hybrid chip cartridge provided in Example 2;

[0259] FIG. 10 is a schematic view showing the back structure of the microfluidic hybrid chip cartridge provided in Example 2;

[0260] FIG. 11 is a schematic side view showing a microfluidic hybrid chip cartridge provided in Example 2;

[0261] FIG. 12 is a schematic view showing a use state of the microfluidic hybrid chip cartridge provided in Example 2;

[0262] FIG. 13 is a schematic diagram of a microfluidic hybrid chip cartridge for generating a microparticle in parallel with high-throughput composed of a plurality of microfluidic hybrid chip cartridges provided in Example 3 in parallel;

[0263] FIG. 14 is a graph showing the continuous stability test results of a lipid nanoparticle prepared by the microfluidic hybrid chip provided in Example 5 of the present invention and a fishbone chip commercially available from a manufacturer PNI;

[0264] FIG. 15 is a comparison of fluorescence intensity of an in vitro transfection of eGFP-LPP prepared by the microfluidic hybrid chip provided in Example 6 of the present invention with that prepared by a fishbone chip commercially available from a manufacturer PNI;

[0265] FIG. 16 is a comparison of the expression levels of GFP proteins of an in vitro transfection of eGFP-LPP prepared by the microfluidic hybrid chip provided in Example 6 of the present invention with that prepared by a fishbone chip commercially available from a manufacturer PNI; and

[0266] FIG. 17 is a schematic diagram of various other curvilinear configurations of mixing cell channels in a particular embodiment.

DETAILED DESCRIPTION OF THE INVENTION

[0267] Preferred embodiments of the present invention will be described in further detail below with reference to the accompanying drawings, and it should be noted that the following embodiments are intended to facilitate understanding of the present invention without any limitation thereto. The raw materials and equipment used in the particular embodiment of the present invention are all known products and are obtained by purchasing commercially available products.

[0268] In the description of the present invention, it is to be understood that the terms “central”, “longitudinal”, “lateral”, “upper”, “lower”, “front”, “rear”, “left”, “right”, “vertical”, “horizontal”, “top”, “bottom”, “inner”, “outer” and the like are used in the orientations and positional relationships indicated in the drawings, which are based on the orientations and positional relationships indicated in the drawings, and are used for convenience in describing the present invention and for simplicity in description, but do not indicate or imply that the device or element so referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus should not be construed as limiting the present invention. Furthermore, the terms “first”, “second”, and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, features defining “first”, “second”, etc. may explicitly or implicitly include one or more such features. In the description of the present invention, unless otherwise specified, the meaning of “a plurality of” means two or more.

[0269] In describing the present invention, it is to be understood that the terms “mounted”, “coupled”, and “connected” are to be interpreted broadly, for example, either fixedly or removable, or integrally, unless expressly specified and limited otherwise, can be mechanically or electrically, directly or indirectly through an intermediary, and may be internally between two elements. The specific meaning of the above terms in the present invention can be understood by a person skilled in the art under specific circumstances.

Example 1 Mixer Provided by the Present Invention

[0270] A schematic diagram of the mixer provided by the Example is shown in FIGS. 5, 6 and 7, wherein FIG. 5 is a schematic diagram showing the structure of the mixer, FIG. 6 is a sectional view A-A of FIG. 5, and FIG. 7 is a schematic diagram showing the flow direction of a sample in the mixer.

[0271] As shown in FIG. 5, the mixer provided by the example includes a mixing unit 1 provided with a first channel 2 being curvilinear and a second channel 3 being rectilinear, which are connected head to end, i.e. connected head to head, and end to end, respectively.

[0272] Preferably, the second channel 3 is semi-circular arc-shaped and has a rectilinear initial segment 4.

[0273] Preferably, a length of the segment 4 is smaller than or equal to ⅓ of a length of the second channel.

[0274] Preferably, the mixer includes two or more mixing units, and each of the mixing units is connected end to end; two adjacent mixing units are a mixing unit A 1 and a mixing unit B 5, the second passage 3 of the mixing unit A is positioned on a right side of the first passage 2, and the second passage 6 of the mixing unit B 5 is positioned on a left side of the first passage 7.

[0275] Preferably, all channel widths 8 are consistent.

[0276] As shown in FIG. 6, preferably, a channel section 9 of the mixer provided by the present invention is rectangular, all the channel section lengths 10 are consistent, and all the channel widths 8 are uniform. The channel section 9 of the mixer can be made in various shapes as desired, such as circular, semi-circular, square, rectangular, triangular, trapezoidal, etc., and for convenience, the channel section of this example is preferably rectangular or square.

[0277] Preferably, the first channel 2 of each of the mixing units communicates in line with the segment 4 of the second channel 6 of the next mixing unit.

[0278] Preferably, the mixer includes six mixing units 1.

[0279] Preferably, the mixing effect can also be further improved by adding more mixing units in series.

[0280] Preferably, a plurality of mixers provided in this example may also be used in series to improve the mixing effect, depending on a need to prepare the product.

[0281] The flow direction of the sample fluid in the mixer is shown in FIG. 7, and the sample flows up and down and is thoroughly mixed in the mixer.

Example 2 Microfluidic Hybrid Chip Cartridge Provided by the Present Invention

[0282] The microfluidic hybrid chip cartridge provided by the Example is shown in FIGS. 8-12, wherein FIG. 8 is a structural schematic diagram of the microfluidic hybrid chi, FIG. 9 is a structural schematic diagram of the microfluidic hybrid chip cartridge with the packaging cartridge, FIG. 10 is a back structural schematic diagram of the microfluidic hybrid chip cartridge with the packaging cartridge, FIG. 11 is a side structural schematic diagram of the microfluidic hybrid chip cartridge with the packaging cartridge, and FIG. 12 is a schematic view showing a state of use of the microfluidic hybrid chip cartridge.

[0283] The microfluidic hybrid chip cartridge provided by the example includes the microfluidic mixer provided by the Example 1.

[0284] As shown in FIGS. 8-11, the example provides a microfluidic hybrid chip cartridge which includes a chip 11 provided thereon with liquid inlets 12 and 312, a liquid outlet 313, liquid inlet conduits 14 and 314, a liquid outlet conduit 15 and a mixer 16, and the liquid inlets 12 and 312 and the liquid outlet 313 are perpendicular to a side wall of the chip; the liquid inlet conduit 14 is connected with the liquid inlet 12 and the mixer 16, the liquid inlet conduit 314 is connected with the liquid inlet 312 and the mixer 16, the liquid outlet conduit 15 is connected with the liquid outlet 313 and the mixer 16, and the packaging cartridge 17 is arranged outside the chip. The liquid inlets 12 and 312 and the liquid outlet 13 are respectively located at either side of the chip 11.

[0285] Preferably, the inlets 12 and 312 and the inlet conduits 14 and 314 are in the same plane, and the liquid outlet 313 and the outlet conduit 15 in the same plane.

[0286] Preferably, the inlets 12 and 312, the inlet conduits 14 and 314, the liquid outlet 313, the outlet conduit 15 and the chip 11 are all substantially in the same plane. The sample is applied by injection only from a side of the chip 11.

[0287] As shown in the FIG. 12, the microfluidic hybrid chip cartridge provided by the present invention has the advantages that liquid inlets 12 and 312 and a liquid outlet 13 are arranged perpendicular to a side wall of a chip 11. When it is used, a syringe is disposed vertically downward for injection, the chip 11 and the syringe are in the same plane, and the syringe is placed vertically downward after it extracts a liquid sample, such that bubbles naturally float to the top inside the syringe, then the syringe is inserted vertically downward into the liquid inlets 12 and 312 of the chip 11, and the liquid in the syringe is completely injected into the liquid inlets 12 and 312. The bubbles float up to the top of the syringe, thus it is not needed to worry about injection of the bubbles, and waste of expensive sample liquid due to manual removal of bubbles at the head of a syringe was avoided.

Example 3 Microfluidic Hybrid Chip Cartridge Provided by the Present Invention for Generating a Microparticle in Parallel with High-Throughput

[0288] As shown in the FIG. 13, the present invention provides a microfluidic mixing cartridge for generating nanoparticles in parallel with high-throughput, which is composed of a plurality of microfluidic hybrid chip cartridges provided in Example 2 in parallel. Due to the fact that the liquid inlets 12 and 312, the liquid outlet 313 and the chip 11 are in the same plane, injection only needs to be carried out from the side surface of the chip 11 during sample application, a plurality of microfluidic hybrid chips can be stacked, thus the microfluidic hybrid chips can be used in parallel with high-throughput and can be used for generating microparticles in parallel with high-throughput.

Example 4 Performance Comparison of Different Chips

[0289] The microfluidic hybrid chip provided by the Example 2 and the commercially available fishbone chip manufactured by PNI are respectively used for preparing a lipid nanoparticle, and the influence of different mixing flow rates on the particle size of the lipid nanoparticle is investigated. To be specific, an appropriate amount of lipid solution (ionizable lipid MC3, DSPC, cholesterol, mPEG2000-DMG prepared into 10 mg/ml lipid solution according to a molar ratio of 50:10:38.5:1.5) is mixed with eGFP-mRNA (dissolved in 1 mM of citric acid-sodium citrate buffer at pH 6.4, mRNA sequence of GFP:

TABLE-US-00001 AUGGUGAGCA AGGGCGAGGA GCUGUUCACC GGGGUGGUGC CCAUCCUGGU CGAGCUGGAC GGCGACGUAA ACGGCCACAA GUUCAGCGUG UCCGGCGAGG 101 GCGAGGGCGA UGCCACCUAC GGCAAGCUGA CCCUGAAGUU CAUCUGCACC ACCGGCAAGC UGCCCGUGCC CUGGCCCACC CUCGUGACCA CCCUGACCUA 201 CGGCGUGCAG UGCUUCAGCC GCUACCCCGA CCACAUGAAG CAGCACGACU UCUUCAAGUC CGCCAUGCCC GAAGGCUACG UCCAGGAGCG CACCAUCUUC 301 UUCAAGGACG ACGGCAACUA CAAGACCCGC GCCGAGGUGA AGUUCGAGGG CGACACCCUG GUGAACCGCA UCGAGCUGAA GGGCAUCGAC UUCAAGGAGG 401 ACGGCAACAU CCUGGGGCAC AAGCUGGAGU ACAACUACAA CAGCCACAAC GUCUAUAUCA UGGCCGACAA GCAGAAGAAC GGCAUCAAGG UGAACUUCAA 501 GAUCCGCCAC AACAUCGAGG ACGGCAGCGU GCAGCUCGCC GACCACUACC AGCAGAACAC CCCCAUCGGC GACGGCCCCG UGCUGCUGCC CGACAACCAC 601 UACCUGAGCA CCCAGUCCGC CCUGAGCAAA GACCCCAACG AGAAGCGCGA UCACAUGGUC CUGCUGGAGU UCGUGACCGC CGCCGGGAUC ACUCUCGGCA 701 UGGACGAGCU GUACAAGUAA),
mixed at different flow rates of 1, 6, 12, and 20 ml/min, fixed mixing ratio of 3 (mRNA solution):1 (lipid solution), constant temperature of 37° C. to obtain a lipid nanoparticle, and a particle size is measured by a dynamic light scattering particle size analyzer and repeated three times, with the results shown in Table 1.

TABLE-US-00002 TABLE 1 Comparison of the particle sizes of lipid nanoparticles made from different chips Sequence Mixing flow rate PNI fishbone Microfluidic hybrid chip number (ml/min) chip (nm) (nm) of Example 1. 1 1 164.7 ± 1.1  156.9 ± 7.9  2 6 88.7 ± 8.1 90.6 ± 5.4 3 12 87.1 ± 4.1 87.2 ± 4.9 4 20 76.7 ± 1.8 83.4 ± 8.5

[0290] As can be seen from table 1, a particle size of the lipid nanoparticle prepared by the chip provided in Example 1 of the present invention is not much different from that of the lipid nanoparticle prepared by a fishbone chip commercially available from a manufacturer PNI within each flow rate range (1, 6, 12, 20 ml/min), but the particle size of the lipid nanoparticle prepared by the chip provided in Example 1 is more stable, and the difference in the particle size is smaller at different flow rates. Therefore, the nanoparticles prepared by the microfluidic hybrid chip provided by the present invention is more uniform and stable, the flow resistance are smaller, the mixing efficiency is higher, production in parallel with high-throughput can also be carried out, and the effect achieved is obviously superior to that achieved by the existing microfluidic chip.

Example 5 Continuous Stability Testing of Chips

[0291] The microfluidic hybrid chip provided by the Example 2 is used for preparing lipid nanoparticles to investigate stability of the chip under continuous mixing preparation. To be specific, an appropriate amount of the lipid solution was mixed with eGFP-mRNA, respectively, at a fixed mixing ratio of 20 ml/min, at a fixed flow rate of mixing 3 (mRNA solution):1 (lipid solution), at a constant temperature of 37° C., mixed for 40 min, points were taken every 10 min to obtain lipid nanoparticles, and a particle size was tested with a dynamic light scattering particle sizer, repeated three times, and the results are as shown in FIG. 14 (composition of lipid solution and eGFP-mRNA refers to Example 4).

[0292] The test results in the FIG. 14 show that the chip structure provided by the present invention is good in continuous stability, the particle size obtained after the chip is continuously operated for 40 minutes is equivalent to an initial value, and the polydispersity index PDI is smaller than 0.05.

Example 6 Fluorescence Imaging and GFP Expression Quantification of eGFP-LPP Prepared by Different Chips

[0293] The microfluidic mixed chip provided in Example 2 and a fishbone chip commercially available from a manufacturer PNI are respectively used for preparing a lipid nanoparticle, a prepared eGFP-LPP is transfected in vitro, fluorescence imaging and GFP expression quantitative results of the eGFP-LPP prepared by different chips are investigated. To be specific, the prepared lipid nanoparticles containing 100 ng of eGFP-mRNA prepared by different chips (containing fluorescent mRNA) were incubated with 2×104 DC2.4 cells for 24 hours, then observed GFP expression of same using a fluorescence microscope, as shown in FIG. 15, and finally GFP expression is quantified using a GFP quantification kit, as shown in FIG. 16 (the composition of the lipid solution and eGFP-mRNA refers to example 4).

[0294] As can be seen in FIG. 15, the test results show that the fluorescence intensity of the in vitro transfection of eGFP-LPP prepared by the chip provided in Example 1 is comparable to that of eGFP-LPP prepared by a fishbone chip commercially available from a manufacturer PNI.

[0295] As shown by the test results of FIG. 16, the expression levels of GFP proteins of an in vitro transfection of eGFP-LPP prepared by the microfluidic hybrid chip provided in Example 1 is comparable to that of the GFP proteins of an in vitro transfection of eGFP-LPP from a fishbone chip commercially available from a manufacturer PNI without much significant difference.

Example 7 Comparison of Mixing Effects of Different Number of Mixing Units

[0296] The microfluidic hybrid chip provided in Example 2 is adopted, wherein the number of the mixing units is 2, 4, 6, 8, and 10, and the lipid nanoparticles are prepared respectively. To be specific, an appropriate amount of the lipid solution is mixed with the eGFP-mRNA, respectively ((composition of the lipid solution and eGFP-mRNA refers to Example 4). Same is continuously mixed at a fixed mixing flow rate of 20 ml/min, a fixed mixing ratio of 3 (mRNA solution):1 (lipid solution), at a fixed temperature of 37° C. for 40 min, points are taken every 10 min to obtain lipid nanoparticles, a particle size was tested by using a dynamic light scattering particle sizer, a dispersion index PDI and encapsulation efficiency were calculated, repeated three times, and the results are as shown in Table 2.

TABLE-US-00003 TABLE 2 Comparison of polymer/mRNA nanoparticle generation using different numbers of units Mixing unit PDI (Dispersion Encapsulation number Particle size Index) efficiency (%) 2 72.4 ± 2.5 0.145 90.3% 4 79.2 ± 3.7 0.112 92.7% 6 83.5 ± 8.5 0.040 99.6% 8 84.1 ± 6.3 0.042 99.4% 10 84.9 ± 5.0 0.043 99.1%

[0297] As can be seen from Table 2, the mixing effect when a mixing pipe using 6 mixing units is used for preparing the nanoparticles can completely satisfy the mixing effect requirement required for preparing nanoparticles.

[0298] Although the present invention is disclosed above, the present invention is not limited thereto. For example, the present invention can be extended according to the application range of the microfluidic field. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention, and therefore, the scope of the present invention should be determined by the scope of the claims.

[0299] The invention shown and described herein may be implemented in the absence of any element or elements, limitation or limitations specifically disclosed herein. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, and it is recognized that various modifications are possible within the scope of the present invention. It is therefore to be understood that, although the present invention has been particularly disclosed by various embodiments and optional features, modifications and variations of the concepts herein described may be resorted to by a person skilled in the art, and that such modifications and variations are considered to fall within the scope of the present invention as defined by the appended claims.

[0300] The contents of the articles, patents, patent applications, and all other documents and electronically available information described or described herein are hereby incorporated by reference in their entirety to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. Applicants hereby incorporate into this application any and all materials and information retained from any such article, patent, patent application or other document.