Disposable membrane-coated exosome purification device

Abstract

The utility model discloses a disposable membrane exosome purification device which comprises a sample bottle, a PBS (Phosphate Buffer Solution) bottle and a purification chip, a first infusion pipeline connected with the sample bottle is provided with a first pinch valve for controlling the connection and disconnection of the first infusion pipeline, and a second infusion pipeline connected with the PBS bottle is provided with a second pinch valve for controlling the connection and disconnection of the second infusion pipeline; a third infusion pipeline connected with the outlet end of the purification chip is provided with a third pinch valve for controlling the connection and disconnection of the third infusion pipeline; a peristaltic pump for providing infusion power is arranged on a fourth infusion pipeline connected with the inlet end of the purification chip; and the sample bottle, the PBS bottle, the purification chip and the matched infusion pipeline form a disposable membrane bag. The exosome purification device solves the problems of complicated operation, high cost, limited treatment capacity, easiness in pollution and the like in the existing exosome purification technology.

Description

TECHNICAL FIELD

[0001] The utility model relates to a one -time membrane package exosome purification device belongs to biological sample purification technical field. BACKGROUND

[0002] Exosome is nanometer -sized vesicle that cell secretes, contains various biological active molecules, has important role in cell -to -cell communication, disease diagnosis and treatment etc. Therefore, it is crucial to purify exosome from biological sample efficiently and accurately.

[0003] At present, common exosome purification technology includes ultrasonic centrifugation (UC), tangential flow filtration (TFF) and chromatography separation (SEC) etc., but these technologies have certain limitations. Ultrasonic centrifugation is complex, time -consuming and energy -consuming, and it is difficult to meet the demand of large -scale sample processing;Tangential flow filtration is prone to membrane pollution problems, resulting in reduced purification efficiency and increased cost;Chromatography separation handles large volume sample slowly, high cost, and cannot adapt to large -scale production.

[0004] In addition, the traditional purification equipment cleaning and maintenance process is complicated, easy to cause cross contamination, affect the purity and quality of exosome. Therefore, it has important practical significance to develop a kind of operation simple, high purification efficiency, low cost and small pollution risk exosome purification device. UTILITY MODEL CONTENT

[0005] Therefore, the utility model provides a one -time membrane package exosome purification device, solves the problems of existing exosome purification technology, such as complicated operation, high cost, limited processing capacity and easy pollution.

[0006] In order to realize the above -mentioned purpose, the utility model provides the following technical scheme: a one -time membrane package exosome purification device, including sample bottle, PBS bottle and purification chip;

[0007] The first liquid transfer line of sample bottle is configured with the first pinch valve for controlling the on-off of the first liquid transfer line, the second liquid transfer line of PBS bottle is configured with the second pinch valve for controlling the on-off of the second liquid transfer line, and the third liquid transfer line connected with the outlet end of the purification chip is configured with the third pinch valve for controlling the on-off of the third liquid transfer line;

[0008] The fourth liquid transfer line connected with the inlet end of the purification chip is configured with the peristaltic pump for providing liquid transfer power;

[0009] The sample bottle, the PBS bottle, the purification chip and the matched liquid transfer line constitute a one -time membrane package.

[0010] As a preferred scheme of a one -time membrane package exosome purification device, the purification chip is provided with a first negative pressure outlet and a second negative pressure outlet;

[0011] The first negative pressure outlet is connected to the female interface of the first electromagnetic valve through a transfusion pipeline;

[0012] The second negative pressure outlet is connected to the female interface of the second electromagnetic valve through a transfusion pipeline.

[0013] As a preferred solution of the disposable membrane-packaged exosome purification device, the first transfusion pipeline is provided with a first optical coupling; the first optical coupling is used to monitor the flow state of the sample in the first transfusion pipeline.

[0014] As a preferred solution of the disposable membrane-packaged exosome purification device, the second transfusion pipeline is provided with a second optical coupling; the second optical coupling is used to monitor the flow state of the PBS buffer in the second transfusion pipeline.

[0015] As a preferred solution of the disposable membrane-packaged exosome purification device, the female interface of the first electromagnetic valve is provided with a first quick connector, and the female interface of the second electromagnetic valve is provided with a second quick connector.

[0016] As a preferred solution of the disposable membrane-packaged exosome purification device, a waste liquid in-machine collection vacuum tank is further provided;

[0017] A third quick connector and a fourth quick connector are provided on the transfusion pipeline between the female interface of the first electromagnetic valve / the female interface of the second electromagnetic valve and the waste liquid in-machine collection vacuum tank, and the third quick connector is connected or disconnected to the fourth quick connector.

[0018] As a preferred solution of the disposable membrane-packaged exosome purification device, a waste liquid out-of-machine recovery vacuum tank is further provided, and the waste liquid out-of-machine recovery vacuum tank is provided with a fifth quick connector and a sixth quick connector;

[0019] When the third quick connector and the fourth quick connector are disconnected:

[0020] The third quick connector and the fifth quick connector are connected, and the fourth quick connector and the sixth quick connector are connected.

[0021] As a preferred solution of the disposable membrane-packaged exosome purification device, an out-of-machine waste liquid barrel is further provided, and the out-of-machine waste liquid barrel and the waste liquid in-machine collection vacuum tank are connected through a transfusion pipeline. A waste liquid pump is provided on the transfusion pipeline between the out-of-machine waste liquid barrel and the waste liquid in-machine collection vacuum tank.

[0022] As a preferred solution of the disposable membrane-packaged exosome purification device, the waste liquid in-machine collection vacuum tank is connected with a vacuum extraction pipeline, and a third electromagnetic valve is provided on the vacuum extraction pipeline.

[0023] The utility model has the advantages that: the peristaltic pump can stably and continuously convey sample and PBS liquid, cooperates accurate negative pressure control, optimizes the flow of target liquid on the ultrafiltration membrane of the purification chip, reduces the membrane pollution and blockage, significantly improves the purification speed; the disposable membrane bag is simple and convenient to install and disassemble, avoids complex cleaning and maintenance operation, reduces human intervention and operation difficulty; the disposable membrane bag can be directly discarded after use, effectively avoids cross contamination, ensures the purity and quality of the exosome; the waste liquid recovery mode can be quickly switched through the quick plug connector, different experimental requirements are met, the flexibility and convenience of the experiment are improved; the low -cost equipment and disposable consumables are adopted, the high -energy centrifugal equipment and expensive chromatography column are avoided, and the production cost is reduced. BRIEF DESCRIPTION OF DRAWINGS

[0024] In order to more clearly illustrate the embodiment of the utility model or the technical scheme in the prior art, the drawings needed in the embodiment or the prior art description will be briefly introduced below. Obviously, the drawings in the following description are only exemplary, and those skilled in the art can also obtain other implementation drawings according to the provided drawings without creating labor.

[0025] Figure 1 The utility model discloses a disposable membrane bag exosome purification device structure schematic view provided in the embodiment thereof;

[0026] Figure 2 The utility model discloses a disposable membrane bag schematic view provided in the embodiment thereof.

[0027] In the drawing, 1, sample bottle;2, PBS bottle;3, purification chip;4, peristaltic pump;5, first pinch valve;6, second pinch valve;7, third pinch valve;8, first electromagnetic valve;9, second electromagnetic valve;10, waste liquid machine in -collection vacuum tank;11, first negative pressure outlet;12, second negative pressure outlet;13, first pair of light coupling;14, second pair of light coupling;15, first quick plug connector;16, second quick plug connector;17, third quick plug connector;18, fourth quick plug connector;19, waste liquid machine -out recovery vacuum tank;20, fifth quick plug connector;21, sixth quick plug connector;22, machine -out waste liquid barrel;23, vacuumizing pipeline;24, third electromagnetic valve;25, waste liquid pump. DETAILED DESCRIPTION

[0028] The embodiments of the present application will be described in detail below with specific examples, and other advantages and effects of the present application can be easily understood by those skilled in the art from the disclosure of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of the present application.

[0029] Referring to Figure 1 and Figure 2 The present embodiment provides a disposable membrane-pack exosome purification device, comprising a sample bottle 1, a PBS bottle 2 and a purification chip 3.

[0030] The first infusion pipeline connected with the sample bottle 1 is provided with a first pinch valve 5 for controlling the on-off of the first infusion pipeline, the second infusion pipeline connected with the PBS bottle 2 is provided with a second pinch valve 6 for controlling the on-off of the second infusion pipeline, and the third infusion pipeline connected with the outlet end of the purification chip 3 is provided with a third pinch valve 7 for controlling the on-off of the third infusion pipeline.

[0031] The fourth infusion pipeline connected with the inlet end of the purification chip 3 is provided with a peristaltic pump 4 for providing infusion power.

[0032] The sample bottle 1, the PBS bottle 2, the purification chip 3 and the matched infusion pipelines constitute a disposable membrane-pack.

[0033] Specifically, by setting the first pinch valve 5, the second pinch valve 6 and the third pinch valve 7, the conveying path and flow of the sample and the PBS buffer can be flexibly controlled. When the sample needs to be conveyed, the first pinch valve 5 is opened, the second pinch valve 6 is closed, and the peristaltic pump 4 is started, so that the sample is pushed by the peristaltic pump 4 along the first infusion pipeline into the purification chip 3; when the PBS buffer needs to be conveyed, the second pinch valve 6 is opened, the first pinch valve 5 is closed, and the PBS buffer enters the purification chip 3. The design of the disposable membrane-pack makes the whole sample processing process more convenient, reduces the possibility of cross contamination, and the membrane-pack can be directly discarded after use without complex cleaning and maintenance.

[0034] In a possible embodiment, the purification chip 3 is provided with a first negative pressure outlet 11 and a second negative pressure outlet 12; the first negative pressure outlet 11 is inserted into the counter interface of a first electromagnetic valve 8 through an infusion pipeline; and the second negative pressure outlet 12 is inserted into the counter interface of a second electromagnetic valve 9 through an infusion pipeline.

[0035] Specifically, during the purification process, the waste liquid machine in-cabin collection vacuum tank 10 establishes a negative pressure environment through the first electromagnetic valve 8 and the second electromagnetic valve 9. When the sample or PBS buffer enters the purification chip 3, under the action of negative pressure, small molecular substances and liquid pass through the ultrafiltration membrane in the purification chip 3, enter the first electromagnetic valve 8 and the second electromagnetic valve 9 through the first negative pressure outlet 11 and the second negative pressure outlet 12 respectively, and then flow into the waste liquid machine in-cabin collection vacuum tank 10. Exosomes and other macromolecular substances are retained in the purification chip 3 due to their size being greater than the pore size of the ultrafiltration membrane, thereby realizing the separation of exosomes and other small molecular impurities and achieving the purpose of purifying exosomes.

[0036] In a possible embodiment, the first infusion pipeline is configured with a first pair of light couplings 13; the first pair of light couplings 13 are used to monitor the sample flow state in the first infusion pipeline; the second infusion pipeline is configured with a second pair of light couplings 14; the second pair of light couplings 14 are used to monitor the PBS buffer flow state in the second infusion pipeline.

[0037] Specifically, the light coupling detects the presence and flow of liquid by emitting and receiving light. When liquid flows through, the light is blocked or reflected, and the light coupling will produce corresponding changes in electrical signals. These signals are transmitted to the control system, which judges the position, flow and flow rate of the liquid according to the signals, and then accurately controls the start and stop of the peristaltic pump 4 and the opening and closing of the pinch valve. For example, when the first pair of light couplings 13 detects that the sample is about to be delivered, the control system can control the peristaltic pump 4 to stop working to prevent idling, and at the same time, the first pinch valve 5 is closed to prevent air from entering the pipeline and affecting the purification process; for the second pair of light couplings 14, similar monitoring and control functions are also achieved when delivering PBS buffer, ensuring that the entire purification process runs accurately according to the predetermined program.

[0038] In a possible embodiment, the first electromagnetic valve 8 is provided with a first quick connector 15 at the butt joint, and the second electromagnetic valve 9 is provided with a second quick connector 16 at the butt joint; the third quick connector 17 and the fourth quick connector 18 are arranged on the infusion pipeline between the butt joint of the first electromagnetic valve 8 / the butt joint of the second electromagnetic valve 9 and the waste liquid machine in-cabin collection vacuum tank 10, and the third quick connector 17 is connected or disconnected to the fourth quick connector 18.

[0039] Specifically, the setting of the quick connector makes the connection and disconnection of the device more convenient and fast. The first quick connector 15 and the second quick connector 16 facilitate the connection and separation between the purification chip 3 and the first electromagnetic valve 8 and the second electromagnetic valve 9, and facilitate the replacement of the disposable membrane package. The third quick connector 17 and the fourth quick connector 18 are used to switch the waste liquid recycling mode. By default, the third quick connector 17 and the fourth quick connector 18 are connected, and the waste liquid flows into the waste liquid machine through the infusion pipeline to collect the vacuum tank 10; when it is needed to switch to the external recycling mode, the third quick connector 17 and the fourth quick connector 18 can be conveniently disconnected, and the pipeline is reconnected to realize the change of the waste liquid recycling mode, thereby improving the flexibility of the use of the device

[0040] In a possible embodiment, a waste liquid machine internal collection vacuum tank 10 is further configured; a waste liquid machine external recycling vacuum tank 19 is further configured, and the waste liquid machine external recycling vacuum tank 19 is provided with a fifth quick connector 20 and a sixth quick connector 21; when the third quick connector 17 and the fourth quick connector 18 are disconnected: the third quick connector 17 and the fifth quick connector 20 are connected, and the fourth quick connector 18 and the sixth quick connector 21 are connected. An external waste liquid barrel 22 is further configured, and the external waste liquid barrel 22 and the waste liquid machine internal collection vacuum tank 10 are connected through an infusion pipeline. The infusion pipeline between the external waste liquid barrel 22 and the waste liquid machine internal collection vacuum tank 10 is provided with a waste liquid pump 25.

[0041] Specifically, when a larger waste liquid storage space is needed or the waste liquid needs to be classified and recycled, the external recycling mode can be switched to. In the external recycling mode, the waste liquid flows into the waste liquid machine external recycling vacuum tank 19 through the connected pipeline. The waste liquid pump 25 between the external waste liquid barrel 22 and the waste liquid machine internal collection vacuum tank 10 is used to transport the waste liquid in the waste liquid machine internal collection vacuum tank 10 to the external waste liquid barrel 22 for centralized treatment. Such a design not only meets the needs of waste liquid treatment in different experimental scenarios, but also ensures that the waste liquid is properly treated and environmental pollution is avoided.

[0042] In a possible embodiment, the waste liquid machine internal collection vacuum tank 10 is connected with an evacuation pipeline 23, and the evacuation pipeline 23 is provided with a third electromagnetic valve 24.

[0043] Specifically, the third electromagnetic valve 24 is used to adjust the vacuum degree of the waste liquid machine internal collection vacuum tank 10. In the purification process, a suitable vacuum degree is crucial for establishing a stable negative pressure environment. By controlling the opening and closing time and degree of the third electromagnetic valve 24, the vacuum degree in the vacuum tank can be accurately adjusted. For example, in the initial stage of sample purification, a higher vacuum degree is needed to quickly separate the exosomes from small molecule substances; and in the washing stage, appropriately reducing the vacuum degree can reduce the pressure on the ultrafiltration membrane of the purification chip 3, thereby prolonging the service life of the membrane. By adjusting the vacuum degree in real time, the efficient and stable progress of the exosome purification process is ensured.

[0044] The assembling process of the utility model is as follows:

[0045] The sample bottle 1 and PBS bottle 2 are respectively installed on the corresponding first infusion pipeline, second infusion pipeline, and ensure that the connection is tight, no leakage. First clamp valve 5, second clamp valve 6, third clamp valve 7 are installed in turn, ensure that its installation position is correct, can normally control the on-off of infusion pipeline. The peristaltic pump 4 is installed on the fourth infusion pipeline, connect the power supply and carry out debugging, ensure that it can stably provide infusion power. The purification chip 3 is installed at the appropriate position, and the interface of the first infusion pipeline, the second infusion pipeline, the third infusion pipeline and the fourth infusion pipeline is connected, and the connection is firm.

[0046] The first electromagnetic valve 8, second electromagnetic valve 9 are installed, the first negative pressure outlet 11, second negative pressure outlet 12 of the purification chip 3 are connected with the docking interface of the first electromagnetic valve 8, second electromagnetic valve 9 through the infusion pipeline respectively, ensure that the sealing is good. The first quick plug connector 15, second quick plug connector 16 are installed at the docking interface of the first electromagnetic valve 8, second electromagnetic valve 9.

[0047] The first pair of light coupling 13 is installed on the first infusion pipeline, the second pair of light coupling 14 is installed on the second infusion pipeline, and it is connected to the control system.

[0048] The waste liquid machine inside collection vacuum tank 10 is connected with the first electromagnetic valve 8, second electromagnetic valve 9 through the infusion pipeline, the third quick plug connector 17, fourth quick plug connector 18 are installed on the infusion pipeline. The waste liquid machine outside recovery vacuum tank 19 is connected with the corresponding infusion pipeline through the fifth quick plug connector 20, sixth quick plug connector 21, ensure that when needed, the waste liquid recovery mode can be conveniently switched. The infusion pipeline between the waste liquid machine outside waste liquid barrel 22 and the waste liquid machine inside collection vacuum tank 10 is connected, and the waste liquid pump 25 is installed. The vacuum pipeline 23 of the waste liquid machine inside collection vacuum tank 10 is connected to the third electromagnetic valve 24, and then connected to the vacuum pump, to ensure that the whole vacuum system is sealed well.

[0049] The working principle of the utility model is as follows:

[0050] First, the prewashing stage: open the second clamp valve 6, close the first clamp valve 5, start the peristaltic pump 4. The PBS buffer solution in the PBS bottle 2 is under the action of the peristaltic pump 4, along the second infusion pipeline into the purification chip 3. At this time, the waste liquid machine inside collection vacuum tank 10 establishes negative pressure through the first electromagnetic valve 8, second electromagnetic valve 9, the PBS buffer solution penetrates the ultrafiltration membrane of the purification chip 3 under the action of negative pressure, and enters the first electromagnetic valve 8, second electromagnetic valve 9 through the first negative pressure outlet 11, second negative pressure outlet 12 respectively, and then flows into the waste liquid machine inside collection vacuum tank 10, to prewash the purification chip 3 and remove possible impurities.

[0051] Second, sample loading stage: close the second pinch valve 6, open the first pinch valve 5, start the peristaltic pump 4. The sample in the sample bottle 1 is pushed by the peristaltic pump 4, and enters the purification chip 3 along the first infusion pipeline. Under the action of negative pressure, small molecules and liquid pass through the ultrafiltration membrane into the waste liquid machine to collect the vacuum tank 10, and the exosome macromolecules are retained in the purification chip 3, realizing the preliminary purification of the exosome. The first pair of light coupling 13 monitors the flow state of the sample in real time, and when the sample is transported, the control system controls the peristaltic pump 4 to stop working, and the first pinch valve 5 is closed.

[0052] Third, cleaning stage: close the first pinch valve 5, open the second pinch valve 6 again, and start the peristaltic pump 4. The PBS buffer enters the purification chip 3 again to clean the ultrafiltration membrane and recover the residual exosome. The cleaned liquid flows into the waste liquid machine to collect the vacuum tank 10 under the action of negative pressure. The second pair of light coupling 14 monitors the flow of the PBS buffer to ensure that the cleaning process proceeds smoothly.

[0053] Fourth, waste liquid recovery mode switching (if needed): if the experiment needs to be switched to the off-machine recovery mode, after the cleaning stage is completed, the third quick connector 17 and the fourth quick connector 18 are disconnected, the third quick connector 17 and the fifth quick connector 20 are connected, and the fourth quick connector 18 and the sixth quick connector 21 are connected. After that, the waste liquid will flow into the off-machine waste liquid recovery vacuum tank 19.

[0054] Fifth, waste liquid discharge stage: when the waste liquid in the waste liquid machine inside the vacuum tank 10 reaches a certain amount, the waste liquid pump 25 is started to transport the waste liquid to the off-machine waste liquid barrel 22 through the infusion pipeline for centralized treatment.

[0055] Sixth, end of purification: after the above steps are completed, the peristaltic pump 4, the pinch valve and the electromagnetic valve are closed, the disposable membrane bag is disassembled and properly handled, the device is simply cleaned, and the next purification operation is prepared.

[0056] Although the utility model has been described in detail above with general description and specific embodiments, some modifications or improvements can be made on the basis of the utility model, which is obvious to those skilled in the art. Therefore, these modifications or improvements made on the basis of not deviating from the spirit of the utility model, all belong to the scope of protection required by the utility model.

Claims

1. A disposable membrane-packed exosome purification device, characterized by, It comprises a sample bottle (1), a PBS bottle (2) and a purification chip (3); The first infusion pipeline connected with the sample bottle (1) is provided with a first pinch valve (5) for controlling the opening and closing of the first infusion pipeline, the second infusion pipeline connected with the PBS bottle (2) is provided with a second pinch valve (6) for controlling the opening and closing of the second infusion pipeline, and the third infusion pipeline connected with the outlet end of the purification chip (3) is provided with a third pinch valve (7) for controlling the opening and closing of the third infusion pipeline. The fourth infusion pipeline connected with the inlet end of the purification chip (3) is provided with a peristaltic pump (4) for providing infusion power. The sample bottle (1), the PBS bottle (2), the purification chip (3) and the matched infusion pipeline constitute a disposable membrane package.

2. A disposable membrane-packed exosome purification device according to claim 1, wherein, The purification chip (3) is provided with a first negative pressure outlet (11) and a second negative pressure outlet (12); The first negative pressure outlet (11) is inserted into the counter interface of the first electromagnetic valve (8) through the infusion pipeline. The second negative pressure outlet (12) is inserted into the counter interface of the second electromagnetic valve (9) through the infusion pipeline.

3. The disposable membrane-packed exosome purification device of claim 1, wherein, The first infusion pipeline is provided with a first pair of light couplings (13), and the first pair of light couplings (13) are used for monitoring the sample flow state in the first infusion pipeline.

4. The disposable membrane-packed exosome purification device of claim 1, wherein, The second infusion pipeline is provided with a second pair of light couplings (14), and the second pair of light couplings (14) are used for monitoring the PBS buffer flow state in the second infusion pipeline.

5. The disposable membrane-packed exosome purification device of claim 2, wherein, The counter interface of the first electromagnetic valve (8) is provided with a first quick plug (15), and the counter interface of the second electromagnetic valve (9) is provided with a second quick plug (16).

6. A disposable membrane-packed exosome purification device according to claim 5, wherein, A waste liquid in-machine collection vacuum tank (10) is further arranged; A third quick plug (17) and a fourth quick plug (18) are arranged on the infusion pipeline between the counter interface of the first electromagnetic valve (8) / the counter interface of the second electromagnetic valve (9) and the waste liquid in-machine collection vacuum tank (10), and the third quick plug (17) is in communication or disconnection with the fourth quick plug (18).

7. A disposable membrane-packed exosome purification device according to claim 6, wherein, A waste liquid out-of-machine recovery vacuum tank (19) is further arranged, and the waste liquid out-of-machine recovery vacuum tank (19) is provided with a fifth quick plug (20) and a sixth quick plug (21); When the third quick plug (17) and the fourth quick plug (18) are disconnected: The third quick plug (17) and the fifth quick plug (20) are in communication, and the fourth quick plug (18) and the sixth quick plug (21) are in communication.

8. A disposable membrane-packed exosome purification device according to claim 7, wherein, An out-of-machine waste liquid tank (22) is further arranged, and the out-of-machine waste liquid tank (22) and the waste liquid in-machine collection vacuum tank (10) are connected through an infusion pipeline, and the infusion pipeline between the out-of-machine waste liquid tank (22) and the waste liquid in-machine collection vacuum tank (10) is provided with a waste liquid pump (25).

9. A disposable membrane-packed exosome purification device according to claim 8, wherein, The waste liquid in-machine collection vacuum tank (10) is connected with an evacuation pipeline (23), and the evacuation pipeline (23) is provided with a third electromagnetic valve (24).

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