A mixer and mixing method

Abstract

The purpose is to provide a mixer which solves the problem of low RNA encapsulation rate of liposomes, comprising a shell and a mixing cavity inside the shell, a first feeding channel and a second feeding channel are arranged on the shell, the first feeding channel and the second feeding channel are communicated with the mixing cavity, the outlet direction of the first feeding channel is staggered with the outlet direction of the second feeding channel, so that the material forms a vortex in the mixing cavity, a discharge channel is arranged at the bottom of the mixing cavity, the discharge channel is communicated with the mixing cavity, and the discharge channel extends to the outer edge of the shell. Since the liposomes and RNA in the high-pressure state are respectively introduced into the first feeding channel and the second feeding channel, the liposomes and RNA rotate in the mixing cavity under the high pressure, and the liposomes and RNA collide and mix in the rotation, so that the liposomes can fully encapsulate the RNA, the encapsulation rate of the liposomes is greatly improved, and the problem of low RNA encapsulation rate of liposomes is solved.

Description

Technical Field

[0001] This invention relates to the field of life science technology, and in particular to a mixer. Background Technology

[0002] RNA and liposomes are usually mixed together using a common three-way structure, which encapsulates the RNA within the liposome, achieving an encapsulation rate of up to 90%.

[0003] Patent publication number CN103906503A discloses a system for the aseptic preparation of lipid-nucleic acid nanoparticles, which is used to collect a concentrated liposome suspension after the removal of organic solvents; wherein the mixing unit contains the drug aqueous solution, and the lipid solution is stably added to the drug solution within the mixing unit for at least 5 minutes to prepare a lipid-drug mixture having a lipid:RNA ratio of up to 12:1.

[0004] To determine the siRNA encapsulation efficiency (EE), expressed as the percentage of siRNA encapsulated in lipid-nucleic acid particles, the following method was used with RiboGreen: This method can be used to determine the concentration of double-stranded and single-stranded RNA or DNA in solution, using instruments including a BioTek Instruments, Inc. FLx800, a variable pipette, and a vortex mixer.

[0005] Although the vortex mixer has been disclosed, the corresponding vortex mixer structure has not been disclosed. Summary of the Invention

[0006] The purpose of this invention is to provide a mixer that solves the problem of low RNA encapsulation rate in liposomes.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] The present invention provides a mixer, comprising a housing and a mixing chamber inside the housing, characterized in that the housing is provided with a first feed channel and a second feed channel, both of which communicate with the mixing chamber, the outlet direction of the first feed channel is offset from the outlet direction of the second feed channel so that the material forms a vortex in the mixing chamber, and a discharge channel is provided at the bottom of the mixing chamber, the discharge channel being connected to the mixing chamber and extending to the outer edge of the housing.

[0009] Optionally, a dome is provided at the top of the mixing chamber, and the dome is located inside the housing.

[0010] Optionally, the first feed channel, the second feed channel, and the discharge channel all have the same inner diameter.

[0011] Optionally, a collection chamber is provided between the mixing chamber and the discharge channel, with both ends of the collection chamber connected to the mixing chamber and the discharge channel, respectively.

[0012] Optionally, the length of the discharge channel is longer than both the first feed channel and the second feed channel.

[0013] Optionally, the material flow rate in the discharge channel is simultaneously faster than the material flow rate in the first feed channel and the material flow rate in the second feed channel.

[0014] Optionally, the outlet extension direction of the first feed channel is consistent with the wall extension direction of the mixing chamber, and the outlet extension direction of the second feed channel is consistent with the wall extension direction of the mixing chamber.

[0015] Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art:

[0016] The mixer of the present invention, in which liposomes and RNA are respectively fed under high pressure in the first feed channel and the second feed channel, both liposomes and RNA rotate in the mixing chamber under high pressure, and collide and mix during rotation, so that the liposomes can fully encapsulate the RNA, thereby greatly improving the encapsulation rate of liposomes and solving the problem of low encapsulation rate of RNA by liposomes.

[0017] Another object of the present invention is to provide a mixing method that makes it easier to encapsulate RNA with liposomes.

[0018] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0019] A hybrid approach:

[0020] S1, high-pressure delivery of liposomes and RNA;

[0021] S2, to create a vortex during the introduction of liposomes and RNA;

[0022] S3. Allow the mixture of liposomes and RNA to enter the discharge channel.

[0023] Optionally, in S1, the high pressure range is 3.6-4.0 MPa.

[0024] Optionally, the mixture of liposomes and RNA is introduced into the discharge channel, which is then further mixed again within the channel, which is sufficiently narrow.

[0025] The mixing method of the present invention solves the problem of insufficient mixing of plasmids and RNA because the plasmids and RNA can be fully mixed in the discharge channel due to the high pressure state and the sufficiently narrow discharge channel. Attached Figure Description

[0026] The following sections will describe some specific embodiments of the invention in detail by way of example and not limitation, with reference to the accompanying drawings. The same reference numerals in the drawings denote the same or similar parts or portions. Those skilled in the art should understand that these drawings are not necessarily drawn to scale. In the drawings:

[0027] Figure 1 This is a perspective view of a mixer according to a preferred embodiment of the present invention;

[0028] Figure 2 yes Figure 1 The diagram shows the internal structure of the mixer.

[0029] Figure 3 yes Figure 1 The image shows a top view of the mixer's internal structure.

[0030] Figure 4 yes Figure 3 The cross-sectional view of AA is shown.

[0031] The reference numerals in the attached figures are explained as follows:

[0032] 1. Shell;

[0033] 2. Mixing chamber;

[0034] 3. First feed channel;

[0035] 4. Second feed channel;

[0036] 5. Discharge channel;

[0037] 6. Dome;

[0038] 7. Collection bin. Detailed Implementation

[0039] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0040] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0041] Furthermore, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0042] like Figure 1 and Figure 2 As shown, the device includes a shell 1 and a mixing chamber 2 inside the shell 1. The shell 1 has a certain thickness and can withstand a pressure of 4.0 MPa. The shell 1 is a block-shaped solid. The shell 1 is provided with a first feed channel 3 and a second feed channel 4. Both the first feed channel 3 and the second feed channel 4 are connected to the mixing chamber 2 and also lead to the outside of the shell 1. The outlet direction of the first feed channel 3 is offset from the outlet direction of the second feed channel 4, meaning that the material coming out of the first feed channel 3 and the material coming out of the second feed channel 4 will not collide or intersect. The materials in this application refer to liposomes and RNA. Liposomes and RNA enter the mixing chamber 2 from the first feed channel 3 and the second feed channel 4, respectively. Only one type of material can enter the first feed channel 3 and the second feed channel 4.

[0043] This is to create a vortex in the mixing chamber 2. Because liposomes and RNA are under high pressure in both the first feed channel 3 and the second feed channel 4, when the liposomes and RNA enter the mixing chamber 2, they do not collide with each other. Instead, they collide with the inner wall of the mixing chamber 2, and then both liposomes and RNA rotate, forming a vortex. In this way, the liposomes and RNA are mixed together. During the mixing process, the liposomes and RNA collide, and the liposomes encapsulate the RNA.

[0044] The bottom of the mixing chamber 2 is provided with a discharge channel 5, which is connected to the mixing chamber 2 and extends to the outer edge of the shell 1. That is, the discharge channel 5 is connected to the outside of the shell 1. The high pressure range in this application is between 3.6MPa and 4.0MPa, and the high pressure in this application reaches 3.8MPa.

[0045] A dome 6 is provided at the top of the mixing chamber 2. The dome 6 is high in the middle and droops around the edges. The dome 6 is located inside the shell 1, which facilitates the rotation of the material.

[0046] The first feed channel 3, the second feed channel 4, and the discharge channel 5 all have the same inner diameter, which ranges from 0.4 mm to 0.6 mm. In this application, the inner diameter of the first feed channel 3, the second feed channel 4, and the discharge channel 5 is all 0.5 mm.

[0047] A collection chamber 7 is provided between the mixing chamber 2 and the discharge channel 5. The two ends of the collection chamber 7 are connected to the mixing chamber 2 and the discharge channel 5 respectively. The inside of the collection chamber 7 is empty, and the outside is an inverted cone structure. The collection chamber 7 is thicker at the top and thinner at the bottom. The collection chamber 7 gradually becomes thinner from top to bottom. After the material is initially mixed, it enters the collection chamber 7. The material falls onto the wall of the collection chamber 7 and slowly slides down before entering the discharge channel 5. The collection chamber 7 is actually a collection channel.

[0048] The length of the discharge channel 5 is longer than both the first feed channel 3 and the second feed channel 4. The length of the discharge channel 5 ranges from 4cm to 6cm, while the lengths of the first feed channel 3 and the second feed channel 4 are between 3cm and 5cm. This length allows RNA and liposomes to mix thoroughly, meaning that RNA and liposomes are further mixed in the discharge channel 5.

[0049] The material flow rate in the discharge channel 5 is faster than the material flow rate in the first feed channel 3 and the second feed channel 4. The material flow rate range in the first feed channel 3 and the second feed channel 4 is between 350 ml / min and 370 ml / min, while the material flow rate range in the discharge channel 5 is between 710 ml / min and 750 ml / min. This prevents the material from becoming congested in the discharge channel 5 and allows the material to flow at a faster speed in the discharge channel 5.

[0050] like Figure 3 and Figure 4 As shown, further, the outlet extension direction of the first feed channel 3 is consistent with the wall extension direction of the mixing chamber 2, and the outlet extension direction of the second feed channel 4 is consistent with the wall extension direction of the mixing chamber 2. In this application, the mixing chamber 2 is a hollow cylinder, so the outlet extension direction of the first feed channel 3 is an arc-shaped surface. The material coming out of the first feed channel 3 starts to rotate along the wall of the mixing chamber 2, and the rotation direction is also circular. The outlet extension direction of the second feed channel 4 is also an arc-shaped surface, and the material coming out of the second feed channel 4 also starts to rotate along the wall of the mixing chamber 2, forming a vortex. During the rotation, the liposomes encapsulate the RNA.

[0051] The first feed channel 3 and the second feed channel 4 are both inclined upwards, and the first feed channel 3 and the second feed channel 4 are inclined in opposite directions. The first feed channel 3 and the second feed channel 4 are symmetrically arranged with respect to the center point of the mixing chamber 2.

[0052] Experimental data show that, using the mixer described in this application, the RNA encapsulation rate of liposomes reaches 98%.

[0053] A mixing method according to the present invention includes:

[0054] S1, high-pressure delivery of liposomes and RNA;

[0055] S2, to create a vortex during the introduction of liposomes and RNA;

[0056] S3. Allow the mixture of liposomes and RNA to enter the discharge channel.

[0057] The high pressure range is 3.6-4.0 MPa. In this application, the high pressure for liposomes and RNA is 3.8 MPa, which is the optimal value.

[0058] The mixture of liposomes and RNA is introduced into the discharge channel. The discharge channel is long and narrow enough that the liposomes and RNA are further mixed. In other words, the liposomes and RNA are first mixed by vortexing. This is a preliminary mixing. Then, the liposomes and RNA enter the long and narrow discharge channel together. Although the liposomes encapsulate the RNA and flow outward, the discharge channel is long enough to allow the liposomes and RNA to be fully mixed again in the discharge channel.

[0059] The above embodiments are only for illustrating the technical concept and features of the present invention. Their purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly. They should not be used to limit the scope of protection of the present invention. All equivalent changes or modifications made in accordance with the spirit and essence of the present invention should be covered within the scope of protection of the present invention.

Claims

1. A mixer comprising a housing (1) and a mixing chamber (2) inside the housing (1), characterized in that The shell (1) is provided with a first feed channel (3) and a second feed channel (4). Both the first feed channel (3) and the second feed channel (4) are connected to the mixing chamber (2). The outlet direction of the first feed channel (3) is offset from the outlet direction of the second feed channel (4) so ​​that the material forms a vortex in the mixing chamber (2). The bottom of the mixing chamber (2) is provided with a discharge channel (5). The discharge channel (5) is connected to the mixing chamber (2). The discharge channel (5) extends to the outer edge of the shell (1). The shell (1) is a block solid. The outlet extension direction of the first feed channel (3) is consistent with the wall extension direction of the mixing chamber (2), and the outlet extension direction of the second feed channel (4) is consistent with the wall extension direction of the mixing chamber (2). Both the first feed channel (3) and the second feed channel (4) are inclined upward. A dome (6) is provided at the top of the mixing chamber (2). The dome (6) is located inside the shell (1). The dome (6) is high in the middle and droops around the edges. A collection chamber (7) is provided between the mixing chamber (2) and the discharge channel (5), and the two ends of the collection chamber (7) are respectively connected to the mixing chamber (2) and the discharge channel (5).

2. A mixer according to claim 1, wherein The first feed channel (3), the second feed channel (4) and the discharge channel (5) all have the same inner diameter, which ranges from 0.4 to 0.6 mm.

3. A mixer according to claim 1, wherein The length range of the first feed channel (3) and the length range of the second feed channel (4) are 3-5cm, and the length range of the discharge channel (5) is 4-6cm.

4. A mixer according to claim 1, wherein The outlet extension direction of the first feed channel (3) is consistent with the wall extension direction of the mixing chamber (2), and the outlet extension direction of the second feed channel (4) is consistent with the wall extension direction of the mixing chamber (2).

5. A mixing method, using the mixer of claim 1, characterized in that, S1, high-pressure delivery of liposomes and RNA; S2, to create a vortex during the introduction of liposomes and RNA; S3. Allow the mixture of liposomes and RNA to enter the discharge channel.

6. A mixing method according to claim 5, wherein, In S1, the high pressure range is 3.6-4.0 MPa.

7. A mixing method according to claim 5, wherein In S2 and S3, the liposomes and RNA are first initially mixed, and then further mixed again in the discharge channel, which is sufficiently narrow and long.

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