An exosome purification system
By combining large and small particle filtration components with a chromatography column purification system, the problem of separating exosomes from other particles in existing technologies has been solved, enabling the acquisition of high-purity exosomes and improving the yield and bioactivity of exosomes.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-18
- Publication Date
- 2026-07-31
AI Technical Summary
In existing technologies, ultrafiltration is difficult to effectively separate exosomes from other particles of the same size, such as apoptotic bodies or microvesicles, resulting in low exosome purity.
A purification system employing a combination of large-particle and small-particle filtration components with a chromatography column removes large and small particulate impurities through circulating filtration and chromatographic separation, ultimately yielding high-purity exosomes.
This method achieves efficient separation and purification of exosomes, improving the purity and yield of exosomes and ensuring the structural integrity and biological activity of exosomes.
Smart Images
Figure CN224578274U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the technical field of biological equipment, and more specifically to an exosome purification system. Background Technology
[0002] Exosomes, as nanoscale (typically 30-150 nm in diameter) extracellular vesicles actively secreted by cells, have shown great potential for application in fields such as intercellular communication, disease diagnosis (e.g., liquid biopsy), drug delivery carriers, and regenerative medicine due to their rich content of bioactive substances such as proteins, lipids, and nucleic acids. With further research, obtaining high-purity, highly active, and structurally intact exosome samples has become a crucial prerequisite for related basic research and clinical applications.
[0003] Currently, the isolation and purification of exosomes mainly rely on ultracentrifugation and ultrafiltration. Ultracentrifugation separates exosomes of different weights by controlling different centrifugation speeds in a centrifuge. However, this method is prone to damaging exosomes, resulting in low exosome yield. Ultrafiltration is a method based on exosome size separation, that is, by designing the diameter of the membrane pores to fit the corresponding size of exosome vesicles. However, since this method is based on size for purification, it is difficult to separate exosomes from other particles of the same size (such as apoptotic bodies or microvesicles). Utility Model Content
[0004] The purpose of this invention is to solve the problem that conventional ultrafiltration methods are difficult to separate exosomes from other particles of the same size (such as apoptotic bodies or microvesicles) when purifying exosomes in the prior art.
[0005] To address the above problems, this utility model provides an exosome purification system, characterized in that it comprises: A large particle filtration assembly includes a first material tank, a first pump body, and a first membrane filter. The first material tank is used to hold raw materials and is connected to the first membrane filter through a first inlet pipe and a first outlet pipe. The filtration pore size of the first membrane filter is larger than the outer diameter of the target material. The first pump body is disposed on the first inlet pipe and is used to transport the raw materials from the first material tank to the first membrane filter. The first outlet pipe is used to transport the raw materials that have not passed through the first membrane filter back to the first material tank. The small particle filtration assembly includes a second material tank, a second pump body, a second membrane filter, a receiving pipeline, and a waste discharge pipeline. The receiving pipeline connects the first membrane filter and the second material tank and is used to transport the raw material filtered by the first membrane filter to the second material tank. The second material tank is connected to the second membrane filter through a second inlet pipe and a second outlet pipe. The pore size of the second membrane filter is smaller than the outer diameter of the target material. The second pump body is located on the second inlet pipe and is used to transport the raw material from the second material tank to the second membrane filter. The second outlet pipe is used to transport the raw material that has not passed through the second membrane filter back to the second material tank. The waste discharge pipeline is used to discharge the raw material that has passed through the second membrane filter to the outside. The purification component includes a chromatography column for receiving raw materials from a second feed tank and performing chromatographic separation.
[0006] Compared with existing technologies, the above solution sets up a large particle filtration component and a small particle filtration component. The large particle filtration component uses a first pump to continuously circulate the sample through a first tank, a first inlet pipe, a first membrane filter, and a first outlet pipe. Exosomes in the sample continuously pass through the first membrane filter and enter the second tank. Subsequently, the second pump of the small particle filtration component continuously circulates the sample through a second tank, a second inlet pipe, a second membrane filter, and a second outlet pipe. Small particulate impurities in the sample are filtered by the second membrane filter and discharged through the waste pipe. Exosomes in the sample, unable to pass through the second membrane filter, return to the second tank through the second outlet pipe, achieving continuous enrichment of the sample in the second tank. Once the enrichment level reaches the required level, the enriched sample in the second tank is further separated by chromatography using a purification component's chromatography column. This further removes impurities of the same size as the target exosomes, ultimately obtaining the target exosomes with excellent purity.
[0007] In an improved embodiment, the purification assembly further includes a purification tank, a transfer pipeline, and a transfer valve. The transfer pipeline connects the second tank and the purification tank and is used to transport the raw material from the second tank to the purification tank. The transfer valve is connected to the transfer pipeline and is used to control the switching of the transfer pipeline. The purification tank delivers the raw material to the chromatography column through the purification pipeline, thereby using the purification tank as an intermediate medium to temporarily store the enriched sample. At this time, the large particle filtration assembly and the small particle filtration assembly can perform cyclic filtration of the next batch of samples, improving efficiency.
[0008] In an improved embodiment, the purification assembly further includes a rinsing tank and a rinsing pump. The purification pipeline is sequentially connected to the purification tank, the rinsing tank, and the chromatography column. The rinsing pump is located in the rinsing tank and is used to deliver eluent into the rinsing tank, thereby rinsing the sample through the rinsing tank and the rinsing pump to facilitate more accurate chromatographic separation of the subsequent chromatography column.
[0009] In an improved embodiment, the purification assembly further includes a pH meter for measuring the pH of the raw material after column chromatography, thereby verifying the purity of the exosome sample obtained after column chromatography separation by pH measurement.
[0010] In an improved embodiment, a temperature control system is also included, which acts on the large particle filtration component, the small particle filtration component, and the purification component to keep the ambient temperature stable at 2–6°C, thereby ensuring stable constant temperature.
[0011] In an improved embodiment, the first inlet pipe is equipped with a first one-way valve directing the flow from the first material tank to the first membrane filter, the second inlet pipe is equipped with a second one-way valve directing the flow from the second material tank to the second membrane filter, and the transfer pipe is equipped with a third one-way valve directing the flow from the second material tank to the purification tank. This design prevents backflow during filtration by the first one-way valve, prevents backflow during filtration by the second one-way valve, and ensures that the sample in the purification tank does not flow back into the second material tank by the third one-way valve.
[0012] In an improved embodiment, a freeze dryer is also included, which receives raw materials from the purification component and freeze-dries them to achieve freeze-drying preservation of the final target exosome sample. Attached Figure Description
[0013] Figure 1 This is a schematic diagram of an exosome purification system.
[0014] Explanation of reference numerals in the attached figures. 1. First material tank; 11. First inlet pipe; 12. First outlet pipe; 13. First check valve; 2. First pump body; 3. First membrane filter; 31. Receiving pipeline; 4. Second material tank; 41. Second inlet pipe; 42. Second outlet pipe; 43. Second check valve; 5. Second pump body; 6. Second membrane filter; 61. Waste discharge pipeline; 7. Chromatography column; 71. Washing tank; 72. Washing pump; 73. pH meter; 8. Purified material tank; 81. Transfer pipeline; 82. Transfer valve; 83. Third check valve; 9. Freeze dryer. Detailed Implementation
[0015] 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.
[0016] 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.
[0017] 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.
[0018] The present application will now be described in further detail with reference to the accompanying drawings and specific embodiments.
[0019] Please see Figure 1 An embodiment of this utility model provides an exosome purification system, comprising: A large particle filtration assembly includes a first material tank 1, a first pump body 2, and a first filter membrane 3. The first material tank 1 is used to hold raw materials. The first material tank 1 is connected to the first filter membrane 3 through a first inlet pipe 11 and a first outlet pipe 12. The filter pore size of the first filter membrane 3 is larger than the outer diameter of the target material. The first pump body 2 is disposed on the first inlet pipe 11 and is used to transport the raw materials of the first material tank 1 to the first filter membrane 3. The first outlet pipe 12 is used to transport the raw materials that have not passed through the first filter membrane 3 back to the first material tank 1. The small particle filtration assembly includes a second material tank 4, a second pump body 5, a second membrane filter 6, a receiving pipeline 31, and a waste discharge pipeline 61. The receiving pipeline 31 connects the first membrane filter 3 and the second material tank 4 and is used to transport the raw material filtered by the first membrane filter 3 to the second material tank 4. The second material tank 4 is connected to the second membrane filter 6 through a second inlet pipe 41 and a second outlet pipe 42. The filtration pore size of the second membrane filter 6 is smaller than the outer diameter of the target material. The second pump body 5 is located in the second inlet pipe 41 and is used to transport the raw material from the second material tank 4 to the second membrane filter 6. The second outlet pipe 42 is used to transport the raw material that has not passed through the second membrane filter 6 back to the second material tank 4. The waste discharge pipeline 61 is used to discharge the raw material that has passed through the second membrane filter 6 to the outside. The purification component includes a chromatography column 7, which is used to receive the raw material from the second feed tank 4 and perform chromatographic separation.
[0020] Compared with existing technologies, the above solution sets up a large particle filtration component and a small particle filtration component. The large particle filtration component uses a first pump 2 to continuously circulate the sample through a first tank 1, a first inlet pipe 11, a first membrane filter 3, and a first outlet pipe 12. The exosomes in the sample continuously pass through the first membrane filter 3 and enter the second tank 4. Subsequently, the second pump 5 of the small particle filtration component continuously circulates the sample through a second tank 4, a second inlet pipe 41, a second membrane filter 6, and a second outlet pipe 42. The small particulate impurities in the sample are filtered by the second membrane filter 6 and discharged by the waste discharge pipe 61. The exosomes in the sample cannot pass through the second membrane filter 6 and return to the second tank 4 from the second outlet pipe 42, achieving continuous enrichment of the sample in the second tank 4. After the enrichment reaches the required level, the enriched sample in the second tank 4 is further separated by chromatography using the chromatography column 7 of the purification component, thereby further removing impurities of the same size as the target exosomes, and finally obtaining the target exosomes with excellent purity.
[0021] In this embodiment, the purification assembly further includes a purification tank 8, a transfer pipeline 81, and a transfer valve 82. The transfer pipeline 81 connects the second tank 4 and the purification tank 8 and is used to transport the raw material from the second tank 4 to the purification tank 8. The transfer valve 82 is connected to the transfer pipeline 81 and is used to control the opening and closing of the transfer pipeline 81. The purification tank 8 transports the raw material to the chromatography column 7 through the purification pipeline. Thus, the purification tank 8 serves as an intermediate medium to temporarily store the enriched sample. At this time, the large particle filtration assembly and the small particle filtration assembly can perform cyclic filtration on the next batch of samples to improve efficiency.
[0022] Furthermore, the purification assembly also includes a rinsing tank 71 and a rinsing pump 72. The purification pipeline is connected in sequence to the purification tank 8, the rinsing tank 71 and the chromatography column 7. The rinsing pump 72 is located in the rinsing tank 71 and is used to deliver the eluent into the rinsing tank 71. Thus, the sample is rinsed through the rinsing tank 71 and the rinsing pump 72, so that the subsequent chromatography separation of the chromatography column 7 is more accurate.
[0023] In addition, the purification assembly may also include a pH meter 73, which is used to measure the pH of the raw material after treatment by the chromatography column 7, thereby verifying the purity of the exosome sample obtained after chromatographic separation by the chromatography column 7 through pH measurement.
[0024] As an optimization of this embodiment, a constant temperature system is also included. The constant temperature system acts on the large particle filtration component, the small particle filtration component, and the purification component to keep the ambient temperature stable at 2-6°C, thereby ensuring stable constantness.
[0025] In this embodiment, the first inlet pipe 11 is provided with a first one-way valve 13 running from the first material tank 1 to the first membrane filter 3, the second inlet pipe 41 is provided with a second one-way valve 43 running from the second material tank 4 to the second membrane filter 6, and the transfer pipe 81 is provided with a third one-way valve 83 running from the second material tank 4 to the purification material tank 8. Thus, the first one-way valve 13 prevents backflow of the first membrane filter 3 during filtration, the second one-way valve 43 prevents backflow of the second membrane filter 6 during filtration, and the third one-way valve 83 ensures that the sample in the purification material tank 8 will not flow back to the second material tank 4.
[0026] As an extension of this embodiment, a freeze dryer 9 is also included. The freeze dryer 9 is used to receive the raw materials provided by the purification component and freeze-dry them, thereby realizing the freeze-drying preservation of the final target exosome sample.
[0027] 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.
[0028] 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.
[0029] 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. An exosome purification system, characterized by, include: A large particle filtration assembly includes a first material tank, a first pump body, and a first membrane filter. The first material tank is used to hold raw materials and is connected to the first membrane filter through a first inlet pipe and a first outlet pipe. The filtration pore size of the first membrane filter is larger than the outer diameter of the target material. The first pump body is disposed on the first inlet pipe and is used to transport the raw materials from the first material tank to the first membrane filter. The first outlet pipe is used to transport the raw materials that have not passed through the first membrane filter back to the first material tank. The small particle filtration assembly includes a second material tank, a second pump body, a second membrane filter, a receiving pipeline, and a waste discharge pipeline. The receiving pipeline connects the first membrane filter and the second material tank and is used to transport the raw material filtered by the first membrane filter to the second material tank. The second material tank is connected to the second membrane filter through a second inlet pipe and a second outlet pipe. The pore size of the second membrane filter is smaller than the outer diameter of the target material. The second pump body is located on the second inlet pipe and is used to transport the raw material from the second material tank to the second membrane filter. The second outlet pipe is used to transport the raw material that has not passed through the second membrane filter back to the second material tank. The waste discharge pipeline is used to discharge the raw material that has passed through the second membrane filter to the outside. The purification component includes a chromatography column for receiving raw materials from a second feed tank and performing chromatographic separation.
2. The exosome purification system of claim 1, wherein, The purification assembly also includes a purification tank, a transfer pipeline, and a transfer valve. The transfer pipeline connects the second tank and the purification tank and is used to transport the raw material from the second tank to the purification tank. The transfer valve is connected to the transfer pipeline and is used to control the opening and closing of the transfer pipeline. The purification tank transports the raw material to the chromatography column through the purification pipeline.
3. The exosome purification system of claim 2, wherein, The purification assembly also includes a rinsing tank and a rinsing pump. The purification pipeline is connected in sequence to the purification tank, the rinsing tank and the chromatography column. The rinsing pump is located in the rinsing tank and is used to deliver the eluent into the rinsing tank.
4. The exosome purification system of claim 3, wherein, The purification assembly also includes a pH meter, which is used to measure the pH of the raw material after it has been treated by the chromatography column.
5. The exosome purification system according to any one of claims 1-4, wherein, It also includes a temperature control system, which acts on the large particle filtration component, the small particle filtration component and the purification component to keep the ambient temperature stable at 2-6°C.
6. The exosome purification system of any one of claims 2-4, wherein, The first inlet pipe is equipped with a first check valve that runs from the first material tank to the first membrane filter, the second inlet pipe is equipped with a second check valve that runs from the second material tank to the second membrane filter, and the transfer pipe is equipped with a third check valve that runs from the second material tank to the purification material tank.
7. The exosome purification system of any one of claims 1-4, wherein, It also includes a freeze dryer, which is used to receive raw materials provided by the purification component and freeze-dry them.