Exosome purification device and method thereof

Abstract

The present invention discloses an exosome purification device and a method thereof, which relate to the technical field of exosome extraction, and include a centrifugal mechanism, a filtering mechanism, and a station switching mechanism arranged between the centrifugal mechanism and the filtering mechanism; the station switching mechanism is respectively provided with a first lifting assembly and a second lifting assembly corresponding to the centrifugal mechanism and the filtering mechanism; the centrifugal mechanism includes a centrifugal bucket, a test tube rack arranged inside the centrifugal bucket and used to store test tube units, a barrel cover matched with the first lifting assembly, and a blocking assembly arranged inside the barrel cover and used to block the pipe opening in the test tube unit; the second lifting assembly is also provided with a liquid taking mechanism, and the liquid taking mechanism is used to absorb the supernatant inside the test tube unit in the centrifugal mechanism and transfer it to the filtering mechanism; the collection result of the present invention has good repeatability, high recovery rate, and greatly improves the efficiency of exosome purification and collection.

Description

Technical Field

[0001] The present invention relates to the technical field related to exosome extraction, and specifically to an exosome purification device and a method thereof. Background Art

[0002] Exosomes are vesicles with a phospholipid bilayer structure secreted by cells. They are between 30-200nm in size and are widely present in plasma, urine, and cell culture fluid. They carry a lot of important protein information and participate in many important biological processes such as intercellular communication and immune response. Due to their small size and origin from the organism itself, they have low toxicity and are non-immunogenic to the organism. Therefore, they have broad prospects for being used as carriers for drug delivery into the body to achieve disease treatment.

[0003] For example, the patent document with patent number CN114134109B discloses a method for purifying EGF mesenchymal stem cell exosomes, comprising the following steps: step 1: centrifuge the cell culture fluid containing EGF mesenchymal stem cell exosomes and take the supernatant; step 2: filter and concentrate the supernatant collected in step 1 with a tangential flow ultrafiltration system to obtain a concentrated solution; step 3: separate and purify the concentrated solution in step 2 with an anion exchange chromatography column and collect the elution peak solution; step 4: separate and purify the elution peak solution collected in step 3 with a heparin affinity chromatography column and collect the elution peak solution to obtain EGF mesenchymal stem cell exosomes. This method can specifically separate and purify EGF mesenchymal stem cell exosomes, and can effectively separate exosomes carrying EGF protein and exosomes not carrying EGF protein. Compared with the traditional ultracentrifugation method, the extracted EGF mesenchymal stem cell exosomes have a higher EGF protein content and can maintain the integrity and biological function of the exosomes.

[0004] However, in actual use, the inventors found that after centrifuging the culture medium, the operator is required to take out the test tubes one by one and then transfer the supernatant to the filtration equipment for filtration. This method is highly dependent on the operator's experience, the collection results have poor reproducibility, the recovery rate is low, and the efficiency of exosome purification and collection is reduced. Summary of the invention

[0005] In order to solve the defects of the prior art, the present invention provides an exosome purification device and a method thereof.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0007] The present invention provides an exosome purification device, comprising:

[0008] A centrifugal mechanism, a filtering mechanism, and a station switching mechanism arranged between the centrifugal mechanism and the filtering mechanism;

[0009] The station switching mechanism is provided with a first lifting assembly and a second lifting assembly corresponding to the centrifugal mechanism and the filtering mechanism respectively;

[0010] The centrifugal mechanism includes a centrifugal bucket, a test tube rack disposed inside the centrifugal bucket and used to store the test tube unit, a bucket cover matched with the first lifting assembly, and a plugging assembly disposed inside the bucket cover and used to plug the tube opening in the test tube unit;

[0011] The second lifting assembly is also provided with a liquid taking mechanism, which is used to absorb the supernatant in the test tube unit in the centrifugal mechanism and transfer it to the filtering mechanism.

[0012] As a preferred technical solution of the present invention, the test tube unit includes a tube body and a test tube plug which cooperates with the tube body and has a through hole in the middle.

[0013] As a preferred technical solution of the present invention, the blocking component includes:

[0014] A rotating disk is rotatably arranged with the barrel cover via a rotating rod;

[0015] A blocking rod is fixedly arranged at the lower end of the rotating disk and is arranged one-to-one corresponding to the through holes;

[0016] The synchronization rods are provided in several groups and are fixedly arranged at the lower end of the rotating disk. The upper end of the test tube rack is provided with several grooves corresponding to the positions of the synchronization rods.

[0017] As a preferred technical solution of the present invention, the first lifting assembly includes:

[0018] A bracket, fixedly arranged on the workstation switching mechanism;

[0019] A threaded rod, rotatably disposed on the bracket and one end of which is transmission-disposed to the first driving unit;

[0020] A fixing plate is fixedly connected to the barrel cover.

[0021] As a preferred technical solution of the present invention, a limiting groove is provided at the upper end of the rotating disk, and a limiting rod is connected to the lower end of the bracket at a position corresponding to the limiting groove through a buffer spring.

[0022] As a preferred technical solution of the present invention, the liquid taking mechanism comprises:

[0023] a second driving unit connected to the second lifting assembly;

[0024] A support plate, fixedly arranged at the lower end of the second lifting assembly and having a plurality of groups of dropper units arranged one-to-one with the test tube units placed therein, and a surrounding plate tightly arranged with the capsule in the dropper unit fixedly arranged at the upper end of the support plate;

[0025] A rotating plate, which is in transmission arrangement with the second driving unit and has a plurality of sliding grooves on its upper end;

[0026] A slide bar is slidably arranged inside the slide groove and a pressing plate is fixedly arranged at the lower end of the slide bar.

[0027] As a preferred technical solution of the present invention, the filtering mechanism comprises:

[0028] A material receiving barrel, the upper end of which is provided with a funnel-shaped receiving plate;

[0029] A driving pump unit, one end of which is connected to the material receiving barrel;

[0030] A filter assembly, wherein a feed pipe is connected to the driving pump unit and a discharge pipe is connected to the receiving barrel;

[0031] The collecting tube is arranged at the lower end of the filtering assembly and is used to collect the enriched liquid.

[0032] As a preferred technical solution of the present invention, the filter assembly includes:

[0033] The outer shell has a side surface on which the feed pipe and the discharge pipe are fixedly arranged via a connecting ring;

[0034] A plurality of filter tubes are arranged side by side and placed inside the outer shell, and both ends of the filter tubes are rotatably arranged with the connecting rings;

[0035] U-shaped tube, used to connect multiple groups of filter tubes;

[0036] A transmission gear is arranged at one end of the filter tube and a plurality of groups of the transmission gears are connected to each other in a transmission manner;

[0037] The driving gear is transmission-connected to one of the transmission gears and is transmission-disposed to the third driving unit.

[0038] As a preferred technical solution of the present invention, the station switching mechanism includes:

[0039] A support platform, on the upper end of which a fourth driving unit is placed;

[0040] A rotating unit, which is arranged in transmission with the fourth driving unit and has a rotating disk fixedly connected to its upper end;

[0041] A rotating wheel, which is arranged on the supporting platform and has an upper end abutting against the rotating disk;

[0042] The connecting rod is used to connect the first lifting assembly and the second lifting assembly to each other.

[0043] The present invention also provides a method for purifying exosomes, comprising the following steps:

[0044] Preparing a cell culture medium containing exosomes;

[0045] Centrifuging the cell culture fluid using a centrifuge mechanism to form a supernatant;

[0046] The liquid taking mechanism sucks the supernatant in the centrifugal mechanism;

[0047] Cooperate with the station switching mechanism to send the supernatant into the interior of the filtering mechanism for filtration;

[0048] The enriched solution obtained from the filtration mechanism is collected.

[0049] The beneficial effects of the present invention are:

[0050] 1. In the present invention, a station switching mechanism is provided to cooperate with a centrifugal mechanism and a filtering mechanism. After the centrifugal mechanism completes the centrifugal work, the station switching mechanism can automatically transfer the supernatant inside the test tube unit in the centrifugal mechanism to the inside of the filtering mechanism without manual operation, so that the collection results have good repeatability and high recovery rate, and the efficiency of exosome purification and collection is greatly improved.

[0051] 2. In the present invention, by arranging the culture fluid to be tangentially filtered inside the filter mechanism, multiple filter tubes can also be rotated synchronously, which is more conducive to some intercepted materials deposited on the filter tubes being washed away as the liquid flows forward, so as to avoid larger intercepted materials from accumulating at the bottom of the filter tube under the action of gravity. Compared with ordinary tangential flow filtration, the filter membrane of the filter tube in the present invention is less likely to be blocked and can be reused. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0053] Figure 1 It is a schematic diagram of the overall structure of the present invention.

[0054] Figure 2 It is a structural schematic diagram of the centrifugal mechanism and the first lifting assembly.

[0055] Figure 3 It is a structural diagram of the centrifugal mechanism.

[0056] Figure 4 It is a schematic diagram of the structure of the plugging component.

[0057] Figure 5 A partial cross-sectional schematic diagram of the plugging component.

[0058] Figure 6 It is a structural schematic diagram of the liquid taking mechanism.

[0059] Figure 7 It is a schematic diagram of the local structure of the liquid taking mechanism.

[0060] Figure 8 It is a structural diagram of the filtering mechanism.

[0061] Fig. 9 A schematic diagram of the structure of the filter component.

[0062] Fig.10 It is a structural schematic diagram of the workstation switching mechanism.

[0063] Fig.11 It is a schematic diagram of the structure of the cleaning mechanism.

[0064] In the figure: 1. centrifugal mechanism; 11. centrifugal bucket; 12. test tube unit; 121. tube body; 122. test tube plug; 1221. through hole; 13. test tube rack; 131. groove; 14. bucket cover; 141. limit sleeve; 15. plugging assembly; 151. rotating disk; 1511. limit groove; 152. rotating rod; 153. plugging rod; 154. synchronous rod; 2. filtering mechanism; 21. receiving barrel; 22. receiving plate; 23. driving pump unit; 24. filtering assembly; 241. outer shell; 242. connecting ring; 243. filter tube; 244. U-shaped tube; 245. transmission gear; 246. driving gear; 247. third driving unit; 25. feeding pipe; 26. discharging pipe; 27. collecting pipe; 3. station switching mechanism; 31. first lifting assembly; 311. bracket; 312. threaded rod; 313. first drive unit; 314. fixing plate; 315. buffer spring; 316. limit rod; 32. second lifting assembly; 33. support platform; 34. fourth drive unit; 35. rotating unit; 36. turntable; 37. rotating wheel; 38. connecting rod; 4. liquid taking mechanism; 41. second drive unit; 42. support plate; 43. dropper unit; 431. capsule; 44. enclosure; 45. rotating plate; 451. slide; 46. slide rod; 47. extrusion plate; 5. cleaning mechanism; 51. cleaning bucket; 52. cleaning water pump; 53. water spray head. DETAILED DESCRIPTION

[0065] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0066] Embodiment 1

[0067] like Figure 1-Figure 8 As shown, an exosome purification device comprises a centrifugal mechanism 1, a filtering mechanism 2 and a station switching mechanism 3 arranged between the centrifugal mechanism 1 and the filtering mechanism 2; the station switching mechanism 3 is respectively provided with a first lifting assembly 31 and a second lifting assembly 32 corresponding to the centrifugal mechanism 1 and the filtering mechanism 2; the centrifugal mechanism 1 comprises a centrifugal bucket 11, a test tube rack 13 arranged inside the centrifugal bucket 11 and used to store a test tube unit 12, a barrel cover 14 matched with the first lifting assembly 31 and a blocking assembly 15 arranged inside the barrel cover 14 and used to block the tube opening in the test tube unit 12; the second lifting assembly 32 is also provided with a liquid taking mechanism 4, and the liquid taking mechanism 4 is used to absorb the supernatant inside the test tube unit 12 in the centrifugal mechanism 1 and transfer it to the filtering mechanism 2.

[0068] Among them, by setting a station switching mechanism 3 to cooperate with the centrifugal mechanism 1 and the filtering mechanism 2, after the centrifugal mechanism 1 completes the centrifugal work, the station switching mechanism 3 can automatically transfer the supernatant inside the test tube unit 12 in the centrifugal mechanism 1 to the inside of the filtering mechanism 2 without manual operation, so that the collection result has good repeatability and high recovery rate, and greatly improves the efficiency of exosome purification and collection, which is particularly suitable for large-scale production of exosomes.

[0069] In detail, during operation, the operator puts the prepared culture solution into the test tube unit 12, and then places the test tube unit 12 on the test tube rack 13 and balances it. At this time, the operator completes the manual processing part, and the subsequent work control equipment can be automatically processed. Specifically, first control the station switching mechanism 3 to drive the first lifting component 31 and the bucket cover 14 to move to the upper end of the centrifugal bucket 11, and then control the first lifting component 31 to drive the bucket cover 14 to descend and close with the centrifugal bucket 11, so that the centrifugal bucket 11 can be controlled to perform centrifugal work;

[0070] After completing the centrifugation work, open the barrel cover 14, and then control the workstation switching mechanism 3 to drive the second lifting component 32 and the liquid taking mechanism 4 to move to the upper end of the centrifuge barrel 11, and then the second lifting component 32 drives the liquid taking mechanism 4 to approach the supernatant of the test tube unit 12 and absorb and transfer this part of the supernatant to the filtering mechanism 2. After the supernatant is transferred to the filtering mechanism 2, the filtering mechanism 2 can filter this part of the supernatant, and the filtered liquid is the required enriched liquid, and finally this part of the enriched liquid can be subjected to subsequent processing.

[0071] Further, if Figure 2-Figure 5 As shown, the test tube unit 12 includes a tube body 121 and a test tube plug 122 matched with the tube body 121 and having a through hole 1221 in the middle;

[0072] The blocking assembly 15 includes a rotating disk 151, a rotating rod 152, a blocking rod 153 and a synchronization rod 154. The rotating disk 151 is rotatably arranged with the barrel cover 14 through the rotating rod 152; the blocking rod 153 is fixedly arranged at the lower end of the rotating disk 151 and is arranged one-to-one corresponding to the through holes 1221; the synchronization rod 154 is provided with several groups and is fixedly arranged at the lower end of the rotating disk 151, and the upper end of the test tube rack 13 is provided with several grooves 131 corresponding to the position of the synchronization rod 154.

[0073] In order to facilitate the collection of the supernatant inside the test tube unit 12, a through hole 1221 is provided on the test tube plug 122. During the centrifugal treatment, the rotating disk 151 drives the blocking rod 153 to extend into the through hole 1221 and block the through hole 1221. At the same time, in order to make the rotating disk 151 stably rotate synchronously with the test tube rack 13, a plurality of grooves 131 are provided on the test tube rack 13. When the blocking rod 153 on the rotating disk 151 extends into the through hole 1221, the synchronous rod 154 arranged at the upper end thereof also extends into the groove 131. The tight fit between the synchronous rod 154 and the groove 131 reduces the pressure applied by the blocking rod 153 to the test tube plug 122.

[0074] When the liquid is being taken, the dropper unit 43 in the liquid taking mechanism 4 can also pass through the through hole 1221 and absorb the supernatant.

[0075] Further, if Figure 4 As shown, the first lifting assembly 31 includes a bracket 311, a threaded rod 312, a first driving unit 313 and a fixed plate 314. The bracket 311 is fixedly arranged on the workstation switching mechanism 3; the threaded rod 312 is rotatably arranged on the bracket 311 and one end thereof is transmission-arranged with the first driving unit 313; the fixed plate 314 is fixedly connected to the barrel cover 14.

[0076] When working, the first driving unit 313 works and drives the threaded rod 312 to rotate. After the threaded rod 312 rotates, it drives the fixing plate 314 to slide on the bracket 311, and then drives the barrel cover 14 to move up and down.

[0077] It should be noted that the structures of the first lifting assembly 31 and the second lifting assembly 32 in the present invention are basically the same, and the difference is that the fixing plate 314 in the second lifting assembly 32 is connected to the second driving unit 41 in the liquid taking mechanism 4.

[0078] Further, if Figure 4-Figure 5 As shown, a limiting groove 1511 is formed at the upper end of the rotating disk 151 , and a limiting rod 316 is connected to the lower end of the bracket 311 at a position corresponding to the limiting groove 1511 through a buffer spring 315 .

[0079] Since the rotating disk 151 is rotatably connected to the barrel cover 14 through the rotating rod 152, when the barrel cover 14 is separated from the centrifugal barrel 11, if the rotating disk 151 rotates randomly, the blocking rod 153 on the rotating disk 151 will not be able to align with the through hole 1221 when the barrel cover 14 is matched with the centrifugal barrel 11 next time. In order to avoid the above situation, a limiting rod 316 is provided at the lower end of the bracket 311 of the first lifting assembly 31. When the rotating disk 151 rises with the barrel cover 14 and the blocking rod 153 is about to be separated from the test tube plug 122, the limiting rod 316 will just extend into To the inside of the limiting groove 1511, and under the action of the buffer spring 315, as the rotating disk 151 continues to rise, the limiting rod 316 will always be placed in the limiting groove 1511. In this way, even if the rotating disk 151 rises, the rotating disk 151 will not rotate at will, thereby ensuring that the blocking rod 153 on the rotating disk 151 can accurately extend into the inside of the test tube plug 122 every time it works (because the test tube rack 13 will stop at the same position every time it stops under the drive of the centrifugal bucket 11, therefore, it is only necessary to control the rotating disk 151 not to rotate at will);

[0080] The limiting rod 316 passes through the barrel cover 14 and extends into the limiting groove 1511 . To ensure that the limiting rod 316 does not deflect, a limiting sleeve 141 may be fixedly provided at the lower end of the barrel cover 14 corresponding to the outer side of the limiting rod 316 .

[0081] Further, if Figure 6-Figure 7 As shown, the liquid taking mechanism 4 includes a second driving unit 41, a support plate 42, a dropper unit 43, a surrounding plate 44, a rotating plate 45, a sliding rod 46 and a squeezing plate 47. The second driving unit 41 is connected to the second lifting assembly 32; the supporting plate 42 is fixedly arranged at the lower end of the second lifting assembly 32 and has a plurality of groups of dropper units 43 arranged one-to-one with the test tube units 12 placed therein, and the upper end of the supporting plate 42 is fixedly provided with a surrounding plate 44 which is tightly arranged with the capsule 431 in the dropper unit 43; the rotating plate 45 is transmission-arranged with the second driving unit 41 and has a plurality of groups of slide grooves 451 opened at the upper end; the sliding rod 46 is slidably arranged inside the slide groove 451 and has a squeezing plate 47 fixedly arranged at the lower end of the sliding rod 46.

[0082] The liquid taking mechanism 4 can be provided to extract the supernatant in the test tube unit 12. Before the dropper unit 43 is extended into the test tube unit 12 under the drive of the second lifting assembly 32, the second driving unit 41 works and drives the rotating plate 45 to rotate. After the rotating plate 45 rotates, the plurality of sliding rods 46 slide in the slide groove 451 and drive the plurality of squeezing plates 47 to move toward the capsule 431 in the dropper unit 43 and squeeze the capsule 431 (the capsule 431 is squeezed on the squeezing plate 47). and the enclosure 44), when the dropper unit 43 extends into the test tube unit 12, the second driving unit 41 works in the reverse direction, thereby driving the multiple squeezing plates 47 to move away from the capsule 431. At this time, the supernatant in the test tube unit 12 will be absorbed by the dropper unit 43. Subsequently, when the dropper unit 43 is transferred to the upper end of the filtering mechanism 2 under the drive of the station switching mechanism 3, the squeezing plates 47 are controlled again to squeeze the capsule 431, so that the supernatant in the dropper unit 43 can be sent to the filtering mechanism 2.

[0083] Further, if Figure 8 As shown, the filtering mechanism 2 includes a material receiving barrel 21, a receiving plate 22, a driving pump unit 23, a filtering assembly 24 and a collecting pipe 27. A funnel-shaped receiving plate 22 is provided at the upper end of the material receiving barrel 21; one end of the driving pump unit 23 is connected to the material receiving barrel 21; the feeding pipe 25 in the filtering assembly 24 is connected to the driving pump unit 23 and the discharging pipe 26 is connected to the material receiving barrel 21; the collecting pipe 27 is provided at the lower end of the filtering assembly 24 and is used to collect the enriched liquid.

[0084] During operation, the supernatant will be sent into the receiving plate 22 by the dropper unit 43, and fall into the receiving bucket 21 along the receiving plate 22. Then, driven by the driving pump unit 23, the supernatant inside the receiving bucket 21 is sent into the filtering component 24 along the feeding pipe 25. After the filtration is completed, the unfiltered part will return to the receiving bucket 21 along the discharging pipe 26 and be filtered again. The filtered part of the exosomes will pass through the filtering component 24 and be discharged from the collecting tube 27.

[0085] Further, if Fig. 9As shown, the filter assembly 24 includes an outer shell 241, a connecting ring 242, a filter tube 243, a U-shaped tube 244, a transmission gear 245, a driving gear 246 and a third driving unit (247, one side of the outer shell 241 is fixedly provided with the feed tube 25 and the discharge tube 26 through the connecting ring 242; a plurality of groups of filter tubes 243 are arranged side by side and placed inside the outer shell 241, and both ends of the filter tubes 243 are rotatably arranged with the connecting ring 242 respectively; the U-shaped tube 244 is used to connect multiple groups of filter tubes 243, the transmission gear 245 is arranged at one end of the filter tube 243 and the multiple groups of the transmission gears 245 are mutually transmission-connected; the driving gear 246 is transmission-connected with one group of the transmission gears 245 and is transmission-arranged with the third driving unit 247.

[0086] Among them, by arranging the culture liquid to be tangentially filtered inside the filter mechanism 2, multiple filter tubes 243 can also rotate synchronously, which is more conducive to some intercepted materials deposited on the filter tube 243 being washed away as the liquid flows forward, so as to avoid larger intercepted materials from accumulating at the bottom of the filter tube 243 under the action of gravity. Compared with ordinary tangential flow filtration, the filter membrane of the filter tube 243 in the present invention is less likely to be blocked and can be reused.

[0087] It should be noted that there are multiple connecting rings 242, which are used to connect the feed pipe 25, the discharge pipe 26 and the multiple U-shaped pipes 244 on the one hand, and are also used to connect with the multiple filter pipes 243 on the other hand;

[0088] During filtering, the third driving unit 247 works and synchronously drives the plurality of transmission gears 245 to rotate through the driving gear 246 , thereby driving the plurality of filter tubes 243 to rotate.

[0089] Further, if Fig.10 As shown, the workstation switching mechanism 3 includes a support platform 33, a fourth driving unit 34, a rotating unit 35, a turntable 36, a rotating wheel 37 and a connecting rod 38. The fourth driving unit 34 is placed on the upper end of the support platform 33; the rotating unit 35 is arranged in a transmission manner with the fourth driving unit 34 and the upper end is fixedly connected with the turntable 36; the rotating wheel 37 is arranged on the support platform 33 and the upper end is abutted against the turntable 36; the connecting rod 38 is used to connect the first lifting assembly 31 and the second lifting assembly 32 to each other.

[0090] When working, the fourth driving unit 34 drives the rotating unit 35 to work, and the rotating unit 35 drives the rotating disk 36 to rotate after working. The rotating disk 36 rotates stably on the multiple rotating wheels 37, and then drives the first lifting assembly 31 and the second lifting assembly 32 to rotate through the connecting rod 38;

[0091] It should be noted that the rotating unit 35 in this embodiment is mainly used to change the power transmission direction of the fourth driving unit 34. Therefore, the rotating unit 35 can be composed of a group of bevel gears, which is a common mechanical structure and its specific structure is not repeated here.

[0092] Embodiment 2

[0093] The components that are the same or corresponding to those in the first embodiment are marked with the same reference numerals as those in the first embodiment. For the sake of simplicity, only the differences from the first embodiment are described below. The second embodiment differs from the first embodiment in that:

[0094] Further, if Fig.11 As shown, a cleaning mechanism 5 is also provided on one side of the workstation switching mechanism 3, which includes a cleaning bucket 51, a cleaning water pump 52 arranged inside the cleaning bucket 51, and a water spray head 53 connected to the cleaning water pump 52 and arranged obliquely upward.

[0095] Among them, by setting up the cleaning mechanism 5, the blocking rod 153 and the dropper unit 43 at the lower end of the barrel cover 14 can be cleaned to facilitate the next use.

[0096] In detail, when cleaning is required, the rotating disk 151 at the lower end of the barrel cover 14 or the rotating plate 45 in the liquid taking mechanism 4 is moved to the upper end of the cleaning barrel 51, and then the rotating disk 151 or the rotating plate 45 and the cleaning barrel 51 are controlled to close by the first lifting assembly 31 or the second lifting assembly 32, so that the cleaning water pump 52 can be controlled to work, and the water flow is sprayed on the blocking rod 153 and the dropper unit 43 through the water spray head 53;

[0097] It should be noted that in order to improve the cleaning efficiency, after using the cleaning mechanism 5 to wash the sealing rod 153 and the dropper unit 43 with water, a drying / air-drying device can also be set near the cleaning mechanism 5 to dry or air-dry the sealing rod 153 and the dropper unit 43. The specific structure will not be repeated here.

[0098] Embodiment 3

[0099] The components identical or corresponding to those in the second embodiment are marked with the same reference numerals as those in the second embodiment. For the sake of simplicity, only the differences from the second embodiment are described below. The third embodiment differs from the second embodiment in that:

[0100] The present invention also provides a method for purifying exosomes, comprising the following steps:

[0101] Preparing a cell culture medium containing exosomes;

[0102] Centrifuge the cell culture fluid using the centrifuge mechanism 1 to form a supernatant;

[0103] The liquid taking mechanism 4 absorbs the supernatant in the centrifugal mechanism 1;

[0104] Cooperate with the station switching mechanism 3 to send the supernatant into the interior of the filtering mechanism 2 for filtration;

[0105] The enriched liquid obtained from the filtering mechanism 2 is collected. It should be noted that the enriched liquid obtained by this method is not the purified exosomes finally obtained, and a chromatography process can be used to further obtain exosomes with higher purity.

[0106] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An exosome purification device, characterized in that: include: A centrifugal mechanism (1), a filtering mechanism (2), and a workstation switching mechanism (3) arranged between the centrifugal mechanism (1) and the filtering mechanism (2); The workstation switching mechanism (3) is provided with a first lifting assembly (31) and a second lifting assembly (32) corresponding to the centrifugal mechanism (1) and the filtering mechanism (2), respectively; The centrifuge mechanism (1) comprises a centrifuge bucket (11), a test tube rack (13) arranged inside the centrifuge bucket (11) and used to store the test tube unit (12), a bucket cover (14) matched with the first lifting assembly (31), and a blocking assembly (15) arranged inside the bucket cover (14) and used to block the tube opening in the test tube unit (12); The second lifting assembly (32) is also provided with a liquid taking mechanism (4), and the liquid taking mechanism (4) is used to suck the supernatant liquid inside the test tube unit (12) in the centrifugal mechanism (1) and transfer it to the filtering mechanism (2); The test tube unit (12) comprises a tube body (121) and a test tube plug (122) matched with the tube body (121) and having a through hole (1221) in the middle; The blocking component (15) comprises: A rotating disk (151) is rotatably arranged with the barrel cover (14) via a rotating rod (152); A blocking rod (153) is fixedly arranged at the lower end of the rotating disk (151) and is arranged in a one-to-one correspondence with the through holes (1221); A plurality of synchronization rods (154) are provided and fixedly arranged at the lower end of the rotating disk (151); a plurality of grooves (131) are provided at the upper end of the test tube rack (13) at positions corresponding to the synchronization rods (154); The first lifting assembly (31) comprises: A bracket (311) fixedly arranged on the workstation switching mechanism (3); A threaded rod (312) rotatably disposed on the bracket (311) and having one end thereof in transmission connection with the first driving unit (313); A fixing plate (314) fixedly connected to the barrel cover (14); The upper end of the rotating disk (151) is provided with a limiting groove (1511), and the lower end of the bracket (311) is connected to a limiting rod (316) via a buffer spring (315) at a position corresponding to the limiting groove (1511); The filtering mechanism (2) comprises: A material receiving barrel (21) having a funnel-shaped receiving plate (22) disposed at its upper end; A driving pump unit (23), one end of which is connected to the material receiving barrel (21); A filter assembly (24), wherein the feed pipe (25) is connected to the drive pump unit (23) and the discharge pipe (26) is connected to the receiving barrel (21); A collecting tube (27), arranged at the lower end of the filtering assembly (24) and used for collecting the enriched liquid; The filtering component (24) comprises: An outer shell (241), one side of which is fixedly provided with the feed pipe (25) and the discharge pipe (26) via a connecting ring (242); A plurality of filter tubes (243) are arranged side by side and placed inside the outer shell (241); both ends of the filter tubes (243) are rotatably arranged with connection rings (242); A U-shaped tube (244) for connecting multiple groups of filter tubes (243); A transmission gear (245) is arranged at one end of the filter tube (243), and a plurality of groups of the transmission gears (245) are in transmission connection with each other; The driving gear (246) is transmission-connected to one of the groups of transmission gears (245) and is transmission-disposed to the third driving unit (247).

2. An exosome purification device according to claim 1, characterized in that: The liquid taking mechanism (4) comprises: A second driving unit (41) connected to the second lifting assembly (32); A support plate (42) is fixedly arranged at the lower end of the second lifting assembly (32) and contains a plurality of groups of dropper units (43) arranged one-to-one with the test tube units (12); a surrounding plate (44) is fixedly arranged at the upper end of the support plate (42) and is closely attached to the capsule (431) in the dropper unit (43); A rotating plate (45) is arranged in transmission with the second driving unit (41) and has a plurality of groups of sliding grooves (451) formed on the upper end; A sliding rod (46) is slidably disposed inside the sliding groove (451), and a pressing plate (47) is fixedly disposed at the lower end of the sliding rod (46).

3. An exosome purification device according to claim 1, characterized in that: The workstation switching mechanism (3) comprises: A support platform (33), with a fourth driving unit (34) placed on the upper end thereof; A rotating unit (35) which is arranged in transmission with the fourth driving unit (34) and has a rotating disk (36) fixedly connected to its upper end; A rotating wheel (37) is arranged on the support platform (33) and the upper end of the rotating wheel is in contact with the rotating disk (36); The connecting rod (38) is used to connect the first lifting assembly (31) and the second lifting assembly (32) to each other.

4. A method for purifying exosomes, comprising the exosome purification device according to any one of claims 1 to 3, characterized in that: The following steps are involved: Preparing a cell culture medium containing exosomes; Centrifuging the cell culture fluid using a centrifuge mechanism (1) to form a supernatant; The liquid taking mechanism (4) takes the supernatant in the centrifugal mechanism (1); Cooperating with the station switching mechanism (3), the supernatant is sent into the interior of the filtering mechanism (2) for filtration; The enriched liquid obtained from the filtering mechanism (2) is collected.

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