Exosome semi-tangential filtration purification device

Abstract

An exosome semi-cutting direction filtration purification device, comprising a filter frame with an inner cavity, and a first filter membrane, a second filter membrane, a raw liquid tank, an enrichment tank and a waste liquid tank, the first filter membrane and the second filter membrane are respectively arranged in the inner cavity and are spaced from each other, so that the inner cavity is divided into a pre-filter cavity, an enrichment cavity and a post-filter cavity; the second filter membrane is inclined from front to back, and the filter pore size of the first filter membrane is larger than that of the second filter membrane; the front side and the rear side of the pre-filter cavity are respectively provided with a liquid inlet pipe and a liquid outlet pipe which are communicated to the raw liquid tank, and the liquid inlet pipe is provided with a pump body for pumping the liquid in the raw liquid tank to the filter frame; the rear side of the enrichment cavity is communicated to the enrichment tank through an enrichment pipe, and the rear side of the post-filter cavity is communicated to the waste liquid tank through a waste discharge pipe. The above scheme can effectively speed up the efficiency of the filtrate and small particle impurities in the sample passing through the second filter membrane and the target exosome being enriched in the enrichment cavity through the inclined arrangement of the second filter membrane, and has good practicability.

Description

Technical Field

[0001] This utility model relates to the technical field of biological purification devices, and more specifically to an exosome semi-tangential filtration purification device. Background Technology

[0002] Exosomes are small membrane vesicles containing complex RNA and proteins. Depending on the research and detection needs, exosomes require purification from samples. The main techniques include centrifugation, polymer precipitation, ultrafiltration, immunoaffinity chromatography, size exclusion chromatography, and ultrafiltration. Ultrafiltration is a method based on exosome size separation. Specifically, it uses the membrane pore size and molecular weight cutoff (MWCO) to allow small particles to pass through the pores while retaining larger particles on the membrane surface. Ultrafiltration is advantageous for processing large volumes of samples, as it allows for the selection of specific exosome sizes by designing the pore diameter. However, its main drawback is that the liquid flow direction is parallel to the pore direction, causing large particles to clog the pores. The resulting shear forces can also deform or break down exosomes.

[0003] To address the aforementioned issues, tangential flow filtration has been proposed. This method changes the primary flow direction of the sample liquid from perpendicular to the filter membrane to parallel to it. This effectively reduces the probability of membrane pore blockage and increases the filter membrane's lifespan while avoiding exosome damage. Tangential flow filtration first uses a filter membrane with a pore size larger than the exosome particle size to remove large particulate impurities from the sample. Then, a filter membrane with a pore size smaller than the exosome particle size removes the filtrate and small particulate impurities from the sample, thus achieving exosome retention and enrichment, resulting in purification. However, because the sample flow direction in tangential flow filtration is parallel to the filter membrane with a smaller pore size, the enrichment efficiency of exosomes on the smaller pore membrane is relatively low. The sample often requires a long period of circulation to achieve exosome enrichment and purification. Utility Model Content

[0004] The purpose of this invention is to solve the problem that in the existing tangential flow filtration method, the sample flow direction is parallel to the filter membrane with a small pore size, resulting in low enrichment efficiency of exosomes on the filter membrane with a small pore size. As a result, the sample often needs to undergo a long period of circulation to achieve enrichment and purification of exosomes.

[0005] To address the aforementioned problems, this utility model provides an exosome semi-tangential filtration purification device, comprising a filter frame with an inner cavity, a first filter membrane, a second filter membrane, a stock solution tank, an enrichment tank, and a waste solution tank. The first and second filter membranes are respectively disposed in the inner cavity from front to back and spaced apart from each other, such that the first filter membrane and one side of the inner cavity enclose a pre-filtration cavity, the second filter membrane and the other side of the inner cavity enclose a post-filtration cavity, and the first and second filter membranes enclose an enrichment cavity. The second filter membrane is inclined from front to back in such a direction that the distance between the second filter membrane and the first filter membrane gradually decreases from front to back, and the pore size of the first filter membrane is larger than that of the second filter membrane.

[0006] The front and rear sides of the pre-filter chamber are respectively provided with an inlet pipe and an outlet pipe connected to the raw liquid tank, and the inlet pipe is provided with a pump body for pumping the liquid from the raw liquid tank to the filter frame; the rear side of the enrichment chamber is connected to the enrichment tank through an enrichment pipe, and the rear side of the post-filter chamber is connected to the waste liquid tank through a waste discharge pipe.

[0007] In the above scheme, the pore size of the first filter membrane is larger than the outer diameter of the target exosome, and the pore size of the second filter membrane is smaller than the outer diameter of the target exosome. The stock solution tank is used to store the sample containing exosomes. When exosome purification is performed, the pump body, combined with the inlet and outlet pipes, realizes the circulation of the sample between the stock solution tank and the pre-filter chamber. The target exosomes in the sample in the pre-filter chamber can pass through the first filter membrane to reach the enrichment chamber, while large particulate impurities are always blocked in the pre-filter chamber. Simultaneously, the above scheme sets a second filter membrane that slopes from front to back in the filter frame. The distance between the membrane and the first filter membrane gradually decreases from front to back, and the enrichment tube is connected to the rear side of the enrichment chamber. Therefore, the sample in the enrichment chamber will flow from front to back. At this time, the sample flow direction in the enrichment chamber will form an angle with the second filter membrane, which greatly accelerates the efficiency of the filtrate and small particulate impurities in the sample passing through the second filter membrane and the enrichment of the target exosomes in the enrichment chamber. When the sample in the enrichment chamber flows to the rear side, relatively pure target exosomes can be obtained. At this time, the enrichment tube will transport the relatively pure target exosomes to the enrichment tank to complete the purification of exosomes.

[0008] In an improved embodiment, an oscillator is also included, which is positioned at the front of the enrichment cavity and is used to emit vibration waves backward, thereby suppressing the scaling effect by emitting vibration waves to the sample in the enrichment cavity.

[0009] In an improved scheme, the angle α between the second filter membrane and the first filter membrane is 5° to 10°. This angle range allows the filtrate and small particulate impurities in the sample to pass through the second filter membrane more quickly, while ensuring that exosomes do not adhere to the second filter membrane and cause blockage.

[0010] In an improved embodiment, the side of the second filter membrane facing the first filter membrane is coated with a negatively charged layer. Since the outer membrane of exosomes is usually negatively charged, by coating the side of the second filter membrane facing the first filter membrane with a negatively charged layer, the adhesion of exosomes to the second filter membrane can be reduced, making it easier for exosomes to flow to the rear side of the enrichment cavity and be discharged from the enrichment tube.

[0011] In an improved embodiment, the front side of the filter frame is provided with a first slot for inserting the front sides of the first filter membrane and the second filter membrane respectively, and the rear side of the filter frame is provided with a second slot for inserting the rear sides of the first filter membrane and the second filter membrane respectively, making installation simple and convenient.

[0012] In an improved embodiment, the pump body is a peristaltic pump, which operates stably and does not affect the sample. Attached Figure Description

[0013] Figure 1 This is a schematic diagram of an exosome semi-tangential filtration and purification device.

[0014] Figure 2 This is a schematic diagram of a semi-tangential filtration and purification device for exosomes, with the upper side of the filter frame hidden.

[0015] Figure 3 This is a top view of a semi-tangential filtration and purification device for exosomes, with the upper side of the filter frame hidden.

[0016] Explanation of reference numerals in the attached figures.

[0017] 1. Filter frame; 11. Pre-filter chamber; 12. Enrichment chamber; 13. Post-filter chamber; 14. First slot; 15. Second slot; 2. First filter membrane; 3. Second filter membrane; 4. Raw material tank; 41. Inlet pipe; 42. Outlet pipe; 43. Pump body; 5. Enrichment tank; 51. Enrichment pipe; 6. Waste liquid tank; 61. Waste discharge pipe; 7. Shaker. Detailed Implementation

[0018] It should be understood by those skilled in the art that the following embodiments are merely illustrative of the technical principles of the embodiments of this application and are not intended to limit the scope of protection of the embodiments of this application. Those skilled in the art can make adjustments as needed to adapt to specific application scenarios.

[0019] In the following description of the embodiments, it should be noted that, unless otherwise explicitly specified and limited, the terms "connected" and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium. Those skilled in the art can understand the specific meaning of the above terms in the embodiments of this application based on the specific circumstances.

[0020] In the embodiments of this application, unless otherwise expressly specified and limited, "above" or "below" the second feature can mean that the first feature is in direct contact with the second feature, or that the first feature is in indirect contact with the second feature through an intermediate medium. Furthermore, "above," "on top of," and "over" the second feature can mean that the first feature is directly above or diagonally above the second feature, or simply that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature can mean that the first feature is directly below or diagonally below the second feature, or simply that the first feature is at a lower horizontal level than the second feature.

[0021] The present application will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0022] Please see Figures 1-3 An embodiment of this utility model provides an exosome semi-tangential filtration and purification device, including a filter frame 1 with an inner cavity, a first filter membrane 2, a second filter membrane 3, a stock solution tank 4, an enrichment tank 5, and a waste solution tank 6. The first filter membrane 2 and the second filter membrane 3 are respectively arranged from front to back in the inner cavity and are spaced apart from each other, so that the first filter membrane 2 and one side of the inner cavity enclose a pre-filtration cavity 11, the second filter membrane 3 and the other side of the inner cavity enclose a post-filtration cavity 13, and the first filter membrane 2 and the second filter membrane 3 enclose an enrichment cavity 12. The second filter membrane 3 is inclined from front to back, and the inclination direction is such that the distance between the second filter membrane 3 and the first filter membrane 2 gradually decreases from front to back, and the filtration pore size of the first filter membrane 2 is larger than the filtration pore size of the second filter membrane 3.

[0023] The front and rear sides of the pre-filtration chamber 11 are respectively provided with an inlet pipe 41 and an outlet pipe 42 connected to the raw liquid tank 4, and the inlet pipe 41 is provided with a pump body 43 for pumping the liquid from the raw liquid tank 4 to the filter frame 1; the rear side of the enrichment chamber 12 is connected to the enrichment tank 5 through an enrichment pipe 51, and the rear side of the post-filtration chamber 13 is connected to the waste liquid tank 6 through a waste discharge pipe 61.

[0024] In the above scheme, the pore size of the first filter membrane 2 is larger than the outer diameter of the target exosome, and the pore size of the second filter membrane 3 is smaller than the outer diameter of the target exosome. The stock solution tank 4 is used to store the sample containing exosomes. When exosome purification is performed, the pump body 43, combined with the inlet pipe 41 and the outlet pipe 42, realizes the circulation of the sample between the stock solution tank 4 and the pre-filter chamber 11. The target exosomes in the sample in the pre-filter chamber 11 can pass through the first filter membrane 2 to reach the enrichment chamber 12, while large particulate impurities are always blocked in the pre-filter chamber 11. At the same time, the above scheme sets a second filter membrane 3 that is inclined from front to back in the filter frame 1. The distance between the filter membrane 3 and the first filter membrane 2 gradually decreases from front to back, and the enrichment tube 51 is connected to the rear side of the enrichment chamber 12. Therefore, the sample in the enrichment chamber 12 will flow from front to back. At this time, the sample flow direction in the enrichment chamber 12 will form an angle with the second filter membrane 3, which greatly accelerates the passage of filtrate and small particulate impurities in the sample through the second filter membrane 3 and the enrichment efficiency of the target exosomes in the enrichment chamber 12. When the sample in the enrichment chamber 12 flows to the rear side, relatively pure target exosomes can be obtained. At this time, the enrichment tube 51 transports the relatively pure target exosomes to the enrichment tank 5, and the purification operation of exosomes can be completed.

[0025] In this embodiment, the filter frame 1 has a cuboid structure, and the interior of the filter frame 1 is a closed cavity. Figure 2 Based on this, the inner cavity of the filter frame 1 is divided from left to right by the first filter membrane 2 and the second filter membrane 3 into a pre-filtration cavity 11, an enrichment cavity 12, and a post-filtration cavity 13.

[0026] As an improvement to this embodiment, an oscillator 7 is also included. The oscillator 7 is disposed on the front side of the enrichment cavity 12 and is used to emit vibration waves backward, thereby emitting vibration waves to the sample in the enrichment cavity 12 through the oscillator 7 to suppress the scaling effect.

[0027] In this embodiment, the angle α between the second filter membrane 3 and the first filter membrane 2 is 5° to 10°. If the angle α is too small, the filtrate and small particulate impurities in the sample will pass through the second filter membrane 3 more slowly; if the angle α is too large, exosomes will easily adhere to the second filter membrane 3 and cause blockage.

[0028] As another improvement to this embodiment, the side of the second filter membrane 3 facing the first filter membrane 2 is coated with a negative charge layer. Since the outer membrane of exosomes is usually negatively charged, by coating the side of the second filter membrane 3 facing the first filter membrane 2 with a negative charge layer, the adhesion of exosomes on the second filter membrane 3 can be reduced, making it easier for exosomes to flow to the rear side of the enrichment cavity 12 and be discharged from the enrichment tube 51.

[0029] In this embodiment, the front side of the filter frame 1 is provided with a first slot 14 for the front sides of the first filter membrane 2 and the second filter membrane 3 to be inserted respectively, and the rear side of the filter frame 1 is provided with a second slot 15 for the rear sides of the first filter membrane 2 and the second filter membrane 3 to be inserted respectively, making installation simple and convenient.

[0030] In this embodiment, the pump body 43 is a peristaltic pump, which operates stably and does not affect the sample.

[0031] It should be noted that in the description of this application, the terms "inner" and "outer," etc., indicating directions or positional relationships, are based on the directions or positional relationships shown in the accompanying drawings. This is only for the convenience of description and does not indicate or imply that the device or component must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation of this application. All directional indications (such as up, down, left, right, front, back, inner, and outer) are only used to explain the relative positional relationships and movement between components in a specific posture. If the specific posture changes, the directional indication will also change accordingly.

[0032] In the description of this application, the references to terms such as "an embodiment," "some embodiments," "in this embodiment," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0033] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A semi-tangential filtration and purification device for exosomes, characterized in that, The system includes a filter frame (1) with an inner cavity, a first filter membrane (2), a second filter membrane (3), a raw liquid tank (4), an enrichment tank (5), and a waste liquid tank (6). The first filter membrane (2) and the second filter membrane (3) are respectively arranged in the inner cavity from front to back and spaced apart from each other, so that the first filter membrane (2) and one side of the inner cavity enclose a pre-filtration cavity (11), the second filter membrane (3) and the other side of the inner cavity enclose a post-filtration cavity (13), and the first filter membrane (2) and the second filter membrane (3) enclose an enrichment cavity (12). The second filter membrane (3) is inclined from front to back, and the inclination direction is such that the distance between the second filter membrane (3) and the first filter membrane (2) gradually decreases from front to back, and the filtration pore size of the first filter membrane (2) is larger than that of the second filter membrane (3). The front and rear sides of the pre-filter chamber (11) are respectively provided with an inlet pipe (41) and an outlet pipe (42) connected to the raw liquid tank (4), and the inlet pipe (41) is provided with a pump body (43) for pumping the liquid from the raw liquid tank (4) to the filter frame (1); the rear side of the enrichment chamber (12) is connected to the enrichment tank (5) through an enrichment pipe (51), and the rear side of the post-filter chamber (13) is connected to the waste liquid tank (6) through a waste discharge pipe (61).

2. The exosome semi-tangential filtration and purification device according to claim 1, characterized in that, It also includes an oscillator (7), which is located on the front side of the enrichment cavity (12) and is used to emit vibration waves backward.

3. The exosome semi-tangential filtration and purification apparatus according to claim 1 or 2, characterized in that, The angle α between the second filter membrane (3) and the first filter membrane (2) is 5° to 10°.

4. The exosome semi-tangential filtration and purification device according to claim 1, characterized in that, The second filter membrane (3) is coated with a negative charge layer on the side facing the first filter membrane (2).

5. The exosome semi-tangential filtration and purification device according to claim 1, characterized in that, The front side of the filter frame (1) is provided with a first slot (14) for the front side of the first filter membrane (2) and the second filter membrane (3) to be inserted respectively, and the rear side of the filter frame (1) is provided with a second slot (15) for the rear side of the first filter membrane (2) and the second filter membrane (3) to be inserted respectively.

6. The exosome semi-tangential filtration and purification apparatus according to claim 1, characterized in that, The pump body (43) is a peristaltic pump.

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