CN217409808U - A high-efficient preparation facilities for exosome - Google Patents
A high-efficient preparation facilities for exosome Download PDFInfo
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- CN217409808U CN217409808U CN202220810800.XU CN202220810800U CN217409808U CN 217409808 U CN217409808 U CN 217409808U CN 202220810800 U CN202220810800 U CN 202220810800U CN 217409808 U CN217409808 U CN 217409808U
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Abstract
The invention discloses a high-efficiency preparation device for exosomes, belonging to the technical field of exosome preparation and comprising the following steps: the chromatography mechanism is used for performing chromatography on substances needing to be removed in the extracellular fluid through the carbon nano tube composite filter layer to obtain an extracellular fluid semi-finished product; a purification unit connected with the chromatography mechanism and used for purifying the external liquid semi-finished product to obtain an exosome; a detection unit; the controller, the high-efficiency preparation device and the preparation method for the exosomes have the advantages that assembly line production is realized, the production efficiency of the exosomes is greatly improved, the consumption of manpower and material resources is reduced, and the occupied area is small; the method is 1/4 of the traditional preparation time, the efficiency is improved by 8 times, the yield is improved by two times, and the engineered collection of exosomes in amniotic membrane stem cells can reach 100ml to prepare 8ml of CB solution. The chromatography effect is improved, and the influence of incomplete chromatography on the purification process is reduced; the real-time data acquisition in the chromatography process is processed by the detection unit through the controller, so that the problem of long production period in the preparation process is solved.
Description
Technical Field
The utility model belongs to the technical field of the exosome preparation, concretely relates to a high-efficient preparation facilities for exosome.
Background
In 1983, exosomes are found in sheep reticulocytes for the first time, and then a plurality of cells can secrete the exosomes under normal and pathological states through research, and the exosomes mainly come from a multivesicular body formed by invagination of intracellular lysosome particles and are released into extracellular matrix after fusion of the multivesicular body outer membrane and cell membranes;
all cultured cell types can also secrete exosomes, and exosomes naturally exist in body fluids including blood, saliva, urine, cerebrospinal fluid and milk, are regarded as specifically secreted membrane vesicles and participate in intercellular communication, and the research on exosomes is increasing, and the function of the exosomes is still understood to be applied to other fields; for example, exosomes play important roles in the pathophysiology such as antigen presentation in immunity, tumor growth and migration, and tissue injury repair, and meanwhile, exosomes secreted by different cells have different components and functions and can be used as biomarkers for disease diagnosis;
the existing methods for extracting exosomes are more, and popular methods comprise an extraction kit, an ultracentrifugation method and the like, wherein the ultracentrifugation method mainly utilizes the difference between the size, the density and the like of the exosomes and substances in the surrounding environment for separation, and has the defects of high requirement on centrifugal equipment, high cost, repeated centrifugation, complex operation steps, large centrifugal force and possibility of damaging the structure of the exosomes, so that the obtained exosomes are low in absorption rate, the extraction kit has the biggest problems of high reagent cost, small extraction amount and incapability of realizing mass production, and the exosomes belong to the laboratory research stage because of the characteristics of multiple types, low content, difficult storage and the like;
therefore, it is necessary to develop an efficient preparation apparatus for extracting biological exosomes to solve the existing problems.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide a high-efficient preparation facilities for exosome to solve the less and long-time problem of output in the exosome preparation process.
In order to achieve the above object, the utility model provides a following technical scheme: a high efficiency production apparatus for exosomes, comprising:
the chromatography mechanism is used for performing chromatography on substances needing to be removed in the extracellular fluid through the carbon nano tube composite filter layer to obtain an extracellular fluid semi-finished product;
a purification unit connected with the chromatography mechanism and used for purifying the external liquid semi-finished product to obtain an exosome;
the detection unit is connected with the chromatographic mechanism and the purification unit and is used for acquiring detection parameters in real time and sending the detection parameters to the controller;
and the controller is connected with the detection unit and used for sending a control instruction.
Preferably, the chromatography mechanism comprises:
the first chromatography tube is internally provided with a carbon nano tube composite filter layer with the aperture of 1400 nm-1200 nm;
the second chromatography tube is internally provided with a carbon nano tube composite filter layer with the aperture of 900nm-1000 nm;
a third chromatography tube, wherein a carbon nano tube composite filter layer with the aperture of 600nm-800 nm is arranged in the third chromatography tube;
the carbon nano tube composite filtering layer is a carbon nano tube composite graphene filtering layer.
Preferably, the detection unit includes:
the bubble detection component is connected with the chromatographic mechanism and is used for detecting the quantity of bubbles flowing through;
the PH detection component is connected with the chromatographic mechanism and is used for acquiring the PH value of the detection body;
the conductivity detection assembly is connected with the chromatographic mechanism and is used for acquiring the conductivity value of the detection body;
the UV detection component is connected with the chromatographic mechanism and used for obtaining a wavelength value according to absorbance by entering incident light with a specific wavelength into the detection body;
a temperature sensor which is arranged in the purification unit and is used for acquiring the temperature value of the detection body;
and the pressure sensor is arranged in the purification unit and is used for acquiring a pressure value.
Preferably, the purification unit comprises:
the first purification mechanism is used for purifying the input external liquid semi-finished product to obtain an exosome with the diameter less than 200 nm;
the second purification mechanism is used for purifying the input external liquid semi-finished product to obtain an exosome with the diameter less than 150 nm;
the flow path selection valve is used for enabling the external liquid semi-finished product to flow into the first purifying mechanism or the second purifying mechanism according to the instruction of the controller;
and the material transferring pump is connected with the flow path selection valve and is used for sucking the external liquid semi-finished product into the input end of the flow path selection valve.
Preferably, the first purifying mechanism comprises:
a first purification tank;
a first spray head connected with the flow path selection valve to spray the external liquid semi-finished product into the first purification box body;
the first purification flow rate sensor is used for acquiring an outflow speed value of exosomes in the first purification box and sending the outflow speed value to the controller;
a first purification carrier having a cut-off value of 500 kD;
a second purification support having a pore size of 200 nm;
a first self-cleaning control valve used for receiving the instruction of the controller and controlling the input of high-pressure gas into the first purifying box body;
the second purifying mechanism includes:
a second purification box;
a second nozzle connected with the flow path selection valve to spray the external liquid semi-finished product into the second purifying box;
the second purification flow rate sensor is used for acquiring the outflow speed value of exosomes in the second purification box and sending the outflow speed value to the controller;
a third purification carrier having a cut-off value of 500 kD;
a fourth purification support having a pore size of 200 nm;
and the second self-cleaning control valve is used for receiving the instruction of the controller and controlling the input of high-pressure gas into the second purifying box body.
Preferably, the first purification box body is of an internal hollow structure, a first spray head is installed at the center of the top of the first purification box body, the distance from the first spray head to the first purification carrier is 2/5 of the height of the whole box body, the distance between the first purification carrier and the second purification carrier is 1/5 of the height of the whole box body, the second purification carrier and the first purification carrier are coaxially distributed, the height from the second purification carrier to the bottom of the second purification carrier is 1/5 of the height of the whole box body, a first purification flow rate sensor is arranged at the bottom of the first purification box body, the first purification flow rate sensor acquires the current outflow speed of the exosome and sends the value to the controller in real time, the controller processes the data of the first purification flow rate sensor and the numerical value of the detection unit and then sends an adjustment instruction to the sample loading pump, and the acquired data of the first purification flow rate sensor is compared with the set parameters, when a trigger condition is met, sending an instruction to a flow path selection valve or a first self-cleaning control valve, and after receiving the instruction, opening a valve by high-pressure gas to sequentially clean a first purification carrier and a second purification carrier from the upper part by the first self-cleaning control valve; the output end of the first self-cleaning control valve is arranged at the top of the first purifying box body, the inner wall of the first purifying box body is provided with a temperature sensor for acquiring the temperature in the box body, and the temperature sensor sends an acquired numerical value to the controller; a pressure sensor for acquiring the pressure of the tank body is arranged at the bottom of the first purifying tank body, and the pressure sensor sends an acquired value to the controller;
the spraying surface of the first spray head is of an arc surface structure, the maximum angle between the arc surface of the first spray head and the horizontal plane is 15 degrees, and the number of spray holes of the spraying surface is 50-80 meshes.
Preferably, the first chromatography tube is of a hollow cylindrical structure, the input end and the output end of the first chromatography tube are both provided with connecting parts for connecting tube plugs, the inner diameter of an opening of each connecting part is 1.2-1.5 times of the inner diameter of the tube body, the wall body of the first chromatography tube is provided with an inverted hook type blocking ring for limiting the carbon nanotube composite graphene, and the distance between the blocking ring and the input end is 2/5 the length of the whole tube body.
Preferably, first chromatography pipe passes through the chromatography support mounting on the installation integrated board, still pass through chromatography support mounting second chromatography pipe and third chromatography pipe on the installation integrated board, just second chromatography pipe, third chromatography pipe, first chromatography pipe are parallel distribution, and with the horizontal plane of installation integrated board is parallel, the internal diameter of first chromatography pipe is 2 times of second chromatography pipe, just the internal diameter of second chromatography pipe is 1.2 times of third chromatography pipe internal diameter, first chromatography pipe, second chromatography pipe, third chromatography pipe highly all the same.
Preferably, the input end of the first chromatography tube is connected with the output end of a sample feeding pump, the sample feeding pump and the pipeline of the first chromatography tube are provided with a bubble detection assembly, the bubble detection assembly sends acquired bubble data to a controller, the controller sends an execution instruction to the sample feeding pump, the output end of the first chromatography tube is provided with a conductance detection assembly and is connected with the input end of a second chromatography tube, the output end of the second chromatography tube is provided with a PH detection assembly, the PH detection assembly sends acquired PH values to the controller, a first flow sensor is arranged between the PH detection assembly and the input end of a third chromatography tube, the output end of the third chromatography tube is connected with a material transferring pump, a UV detection assembly is arranged between the material transferring pump and the third chromatography tube, and sends wavelength values in liquid to the controller, the material transferring pump is connected with the flow path selection valve;
and the output end of the flow path selection valve is provided with a second flow sensor, and the second flow sensor sends a pipeline flow speed value to the controller.
The utility model discloses a technological effect and advantage: the efficient preparation device for the exosome realizes flow line production, greatly improves the production efficiency of the exosome, reduces the consumption of manpower and material resources and occupies less land; 1/4, the efficiency is improved by 8 times, the yield is improved by two times, and 8ml CB solution can be prepared when the engineered collection of the amniotic membrane stem cells reaches 100 ml; the method has the following advantages:
1. the first chromatography tube, the second chromatography tube and the third chromatography tube are used for gradually carrying out chromatography on macromolecules, so that the chromatography effect is improved, and the influence of incomplete chromatography on a purification process is reduced;
2. the carbon nano tube composite graphene filtering layers with different specifications are arranged by the first chromatography tube, the second chromatography tube and the third chromatography tube, the graphene and the carbon nano tube have very good physical and chemical properties, and due to the synergistic effect between the graphene and the carbon nano tube, the electrical conductivity, the mechanical property and other properties of the graphene and carbon nano tube composite material are enhanced, and the chromatography speed is improved;
3. the real-time data in the chromatographic process is acquired by the detection unit and processed by the controller, so that the problem of long production period in the preparation process is solved;
4. different purification processes are carried out on the semi-finished product through the flow path selection valve, so that the purification speed is improved;
5. the purity of the preparation is improved by the arrangement of the first purification carrier and the second purification carrier;
6. through first self-cleaning control valve, solved the jam of purification working process and caused the purification effect to reduce, used high-pressure gas to realize the self-cleaning.
Drawings
Fig. 1 is a schematic structural view of the present invention;
fig. 2 is a front view of the present invention;
FIG. 3 is a right side view of the present invention;
fig. 4 is a left side view of the present invention;
fig. 5 is a structural diagram of the first purifying mechanism and the second purifying mechanism of the present invention without a cover plate;
FIG. 6 is a schematic view of the connection of the electric components of the present invention;
FIG. 7 is a flow chart of the preparation method of the present invention.
In the figure: 11. a first chromatography tube; 12. a second chromatography tube; 13. a third chromatography tube; 14. a sample loading pump; 15. a first chromatography pump; 16. a second chromatography pump; 20. a flow path selection valve; 21. a first purification mechanism; 22. a second purification mechanism; 23. a material transferring pump; 30. a chromatography rack; 40. a connecting portion; 50. a controller; 51. an interactive terminal; 61. a bubble detection assembly; 62. a pH detection component; 63. a conductance detection component; 64. a UV detection component; 65. a pressure sensor; 66. a temperature sensor; 67. a first flow sensor; 68. a second flow sensor; 211. a first purification box; 212. a first nozzle; 213. A first purification vector; 214. a second purification vector; 215. a first self-cleaning control valve; 216. a first purification flow rate sensor; 223. A third purification vector; 224. a fourth purification vector; 225. a second purification flow rate sensor; 226. a second self-purging control valve.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
The utility model provides a high-efficient preparation facilities for exosome as shown in figure 1, include:
the chromatography mechanism is used for separating out substances to be removed from the extracellular fluid through the carbon nano tube composite graphene layer to obtain an extracellular fluid semi-finished product; the chromatography mechanism comprises:
the first chromatography tube 11 is provided with a carbon nano tube composite graphene filter layer with the aperture of 1400-1200 nm;
the second chromatography tube 12 is internally provided with a carbon nano tube composite graphene filtering layer with the aperture of 900nm-1000 nm;
the third chromatography tube 13 is internally provided with a carbon nano tube composite graphene filter layer with the aperture of 600nm-800 nm; the macromolecule is gradually chromatographed, so that the chromatography effect is improved, and the influence of incomplete chromatography on the purification process is reduced; the arranged carbon nanotube composite graphene chromatography bodies with different specifications improve the chromatography speed;
it is worth noting that the first chromatography tube 11 is a hollow cylinder structure, the input end and the output end of the first chromatography tube are both provided with a connecting part 40 for connecting a tube plug, the inner diameter of an opening of the connecting part 40 is 1.2-1.5 times of the inner diameter of the tube body, the wall body of the first chromatography tube 11 is provided with a barb type blocking ring for limiting the carbon nanotube composite graphene, and the length of the blocking ring from the input end is 2/5 of the length of the whole tube body;
as shown in fig. 2, 3 and 4, the first chromatography tube 11 is mounted on a mounting integrated plate through a chromatography bracket 30, a second chromatography tube 12 and a third chromatography tube 13 are further mounted on the mounting integrated plate through the chromatography bracket 30, the second chromatography tube 12, the third chromatography tube 13 and the first chromatography tube 11 are distributed in parallel and parallel to the horizontal plane of the mounting integrated plate, the inner diameter of the first chromatography tube 11 is 2 times that of the second chromatography tube 12, the inner diameter of the second chromatography tube 12 is 1.2 times that of the third chromatography tube 13, and the heights of the first chromatography tube 11, the second chromatography tube 12 and the third chromatography tube 13 are all the same;
the input end of the first chromatographic tube 11 is connected with the output end of the sample loading pump 14, the sample loading pump 14 and the pipeline of the first chromatographic tube 11 are provided with a bubble detection component 61, the bubble detection component 61 sends acquired bubble data to the controller 50, the controller 50 sends an execution instruction to the sample loading pump 14, the output end of the first chromatographic tube 11 is provided with a conductance detection component 63 and is connected with the input end of the second chromatographic tube 12, the output end of the second chromatographic tube 12 is provided with a PH detection component 62, the PH detection component 62 sends acquired PH values to the controller 50, a first flow sensor 67 is arranged between the PH detection component 62 and the input end of the third chromatographic tube 13, the output end of the third chromatographic tube 13 is connected with the material transferring pump 23, a UV detection component 64 is arranged between the material transferring pump 23 and the third chromatographic tube 13, and the UV detection component 64 sends the wavelength values in the liquid to the controller 50, the material transferring pump 23 is connected with the flow path selection valve 20;
wherein, the output end of the flow path selection valve 20 is provided with a second flow sensor 68, and the second flow sensor 68 sends a pipeline flow rate value to the controller 50;
in this embodiment, a first chromatography pump 15 is further disposed on the pipeline between the first chromatography tube 11 and the second chromatography tube 12 to increase the chromatography speed; a second chromatography pump 16 is arranged between the second chromatography tube 12 and the third chromatography tube 13 to improve the speed of chromatography; the first chromatography pump 15 and the second chromatography pump 16 are both connected with the controller 50;
as shown in fig. 5, the purification unit is connected to the chromatography mechanism for purifying the external liquid semi-finished product to obtain exosome; the purification unit comprises:
the first purification mechanism 21 is used for purifying the input external liquid semi-finished product to obtain an exosome with the diameter less than 200 nm; the first purifying mechanism 21 includes:
a first purification tank 211;
a first nozzle 212 connected to the flow path selection valve 20 for spraying the external liquid semi-finished product into the first purification box 211; the flow path selection valve 20 carries out different purification processes on the semi-finished product, so that the purification speed is increased;
a first purification flow rate sensor 216, configured to obtain an outflow rate value of exosomes in the first purification tank 211 and send the outflow rate value to the controller 50;
a first purification carrier 213, said first purification carrier 213 having a cut-off value of 500 kD;
a second purification support 214, the pore size of the second purification support 214 being 200 nm;
a first self-purging control valve 215 for receiving a command from the controller 50 to control the input of the high pressure gas into the first purification tank 211; the problem that the purification effect is reduced due to blockage in the purification working process is solved, and self-cleaning is realized by using high-pressure gas;
the first purifying box 211 is an internal hollow structure, the top center thereof is provided with a first spray head 212, the distance from the first spray head 212 to the first purifying carrier 213 is 2/5 of the whole box height, the distance between the first purifying carrier 213 and the second purifying carrier 214 is 1/5 of the whole box height, the second purifying carrier 214 and the first purifying carrier 213 are coaxially distributed, the distance from the second purifying carrier 214 to the bottom is 1/5 of the whole box height, the bottom of the first purifying box 211 is provided with a first purifying flow rate sensor 216, the first purifying flow rate sensor 216 acquires the current excretion speed and sends the value to the controller 50 in real time, the controller 50 processes the data of the first purifying flow rate sensor 216 and the value of the detecting unit and then sends an adjusting instruction to the sample pump 14, and the acquired data of the first purification flow rate sensor 216 is compared with the set parameters, and when the trigger condition is reached, an instruction is sent to the flow path selection valve 20 or the first self-cleaning control valve 215, and the first self-cleaning control valve 215 opens the valve after receiving the instruction, and the high-pressure gas sequentially cleans the first purification carrier 213 and the second purification carrier 214 from the upper part; the output end of the first self-cleaning control valve 215 is arranged at the top of the first purifying box 211, a temperature sensor 66 for acquiring the temperature in the box is arranged on the inner wall of the first purifying box 211, and the temperature sensor 66 sends the acquired value to the controller 50; and the bottom of the first purifying tank 211 is provided with a pressure sensor 65 for acquiring the tank pressure, and the pressure sensor 65 sends the acquired value to the controller 50, as shown in fig. 6;
the spraying surface of the first nozzle 212 is of an arc surface structure, the maximum angle between the arc surface and the horizontal plane is 15 degrees, and the number of spray holes of the spraying surface is 50-80 meshes;
the second purification mechanism 22 is used for purifying the input external liquid semi-finished product to obtain an exosome with the diameter less than 150 nm; the second purifying mechanism 22 comprises
A second purification box;
a second nozzle; connected with the flow path selection valve 20 to spray the external liquid semi-finished product into the second purification box body; the structure is the same as that of the first nozzle 212;
a second purification flow rate sensor 225, configured to obtain an outflow rate value of exosomes in the second purification tank and send the outflow rate value to the controller 50;
a third purification carrier 223, said third purification carrier 223 having a cut-off value of 500 kD;
a fourth purification support 224, the pore size of the fourth purification support 224 being 200 nm;
a second self-cleaning control valve 226 for receiving the command from the controller 50 to control the input of the high-pressure gas into the second purification tank
A flow path selection valve 20 for making the external liquid semifinished product flow into the first purifying mechanism 21 or the second purifying mechanism 22 according to the instruction of the controller 50; the first purification mechanism 21 or the second purification mechanism 22 works in the same principle, except that the second purification mechanism 22 does not use the second purification carrier 214, and is changed into a fourth purification carrier 224, and the pore diameter of the fourth purification carrier 224 is 200 nm;
and the material transferring pump 23 is connected with the flow path selection valve 20 and is used for sucking the external liquid semi-finished product into the input end of the flow path selection valve 20.
The detection unit is connected with the chromatography mechanism and the purification unit, acquires detection parameters in real time and sends the detection parameters to the controller 50; the detection unit includes:
the bubble detection component 61 is connected with the chromatographic mechanism and is used for acquiring the number of bubbles flowing through;
a PH detection component 62 connected with the chromatography mechanism and used for acquiring the PH value of the detected body;
the conductivity detection assembly 63 is connected with the chromatographic mechanism and is used for acquiring the conductivity value of the detection body;
the UV detection component 64 is connected with the chromatographic mechanism and is used for obtaining a wavelength value according to absorbance when incident light with a specific wavelength enters the detection body;
a temperature sensor 66 provided in the purification unit for acquiring a temperature value of the detection body;
and a pressure sensor 65 arranged in the purification unit for acquiring a pressure value.
The controller 50 is a data processing device with an artificial intelligence processor, and the controller 50 is a data processing device with a storage and networking interface, and is used for analyzing and processing the acquired detection parameters, comparing the acquired detection parameters with set process parameters, and then sending a control instruction, in this embodiment, the controller is connected with the interactive terminal 51; the interactive terminal 51 is used for inputting a process flow and is provided with a communication interface to be connected with a PC;
the utility model discloses the theory of operation, as shown in fig. 7, including following step:
step 1, sucking extracellular fluid into a first chromatographic tube 11 through a sample loading pump 14, acquiring a flowing bubble numerical value by a bubble detection assembly 61 during sucking and sending the flowing bubble numerical value to a controller 50, carrying out chromatography on larger impurities by a carbon nanotube composite graphene filtering layer with the aperture of 1400-1200 nm in the first chromatographic tube 11 and then flowing into a conductance detection assembly 63, acquiring a conductance numerical value by the conductance detection assembly 63 and sending the conductance numerical value to the controller 50;
step 2, the extracellular fluid treated in the step 1 flows into a second chromatography tube 12, a carbon nanotube composite graphene filtering layer with the aperture of 900nm-1000 nm in the second chromatography tube 12 flows into a PH detection component 62 after cell fragments and micro-vesicles are removed by chromatography, after a PH value is obtained, a first flow sensor 67 arranged at the output end of the PH detection component 62 obtains real-time flow information and sends the real-time flow information to a controller 50, and the controller 50 adjusts the power of a sample loading pump 14 according to the bubble value, the conductance value, the PH value and flow data;
step 3, the extracellular fluid treated in the step 2 enters a carbon nano tube composite graphene filtering layer with the aperture of 600nm-800 nm in the third chromatography tube 13 to filter apoptotic bodies and flow into the UV detection assembly 64, if the UV detection assembly 64 detects that the wavelength is less than 400nm, the flow path selection valve 20 is switched to the first purification box body 211, otherwise, the flow path selection valve is switched to the second purification box body;
step 4, flowing into the first purification box 211, passing through the first purification carrier 213 with cut-off value of 500kD, removing small particles, for example, flowing free protein into the second purification carrier 214 with pore size of 200 nm; if the small particles are removed by passing through the third purification carrier 223 with a cut-off value of 500kD after flowing into the second purification box, for example, the free protein flows into the fourth purification carrier 224 with a pore size of 150nm, and then the exosome can be obtained;
the utility model realizes the flow line production, greatly improves the production efficiency of exosomes, reduces the consumption of manpower and material resources and occupies less land; the method is 1/4 of the traditional preparation time, the efficiency is improved by 8 times, the yield is improved by two times, and the engineered collection of exosomes in amniotic membrane stem cells can reach 100ml to prepare 8ml of CB solution.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions on some technical features, and any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of the present invention.
Claims (9)
1. A high-efficient preparation facilities for exosome which characterized in that: the method comprises the following steps:
the chromatography mechanism is used for performing chromatography on substances needing to be removed in the extracellular fluid through the carbon nano tube composite filter layer to obtain an extracellular fluid semi-finished product;
a purification unit connected with the chromatography mechanism and used for purifying the external liquid semi-finished product to obtain an exosome;
the detection unit is connected with the chromatography mechanism and the purification unit and is used for acquiring detection parameters in real time and sending the detection parameters to the controller (50);
and a controller (50) connected to the detection unit for sending control instructions.
2. A high efficiency production apparatus for exosomes according to claim 1, characterised in that: the chromatography mechanism comprises:
the device comprises a first chromatographic tube (11), wherein a carbon nano tube composite filter layer with the aperture of 1400-1200 nm is arranged in the first chromatographic tube (11);
the second chromatography tube (12), the second chromatography tube (12) is internally provided with a carbon nano tube composite filter layer with the aperture of 900nm-1000 nm;
a third chromatography tube (13), wherein a carbon nano tube composite filter layer with the aperture of 600nm-800 nm is arranged in the third chromatography tube (13);
the carbon nano tube composite filtering layer is a carbon nano tube composite graphene filtering layer.
3. A high efficiency production apparatus for exosomes according to claim 1, characterised in that: the detection unit includes:
a bubble detecting unit (61) connected to the chromatographic mechanism for detecting the amount of bubbles flowing therethrough;
a pH detection module (62) connected to the chromatographic mechanism for obtaining a pH value of the specimen;
a conductivity detection module (63) connected to the chromatographic mechanism for acquiring a conductivity value of the detection object;
the UV detection component (64) is connected with the chromatographic mechanism and is used for obtaining a wavelength value according to the absorbance when the incident light with a specific wavelength enters the detection body;
a temperature sensor (66) which is arranged in the purification unit and is used for acquiring the temperature value of the detection body;
a pressure sensor (65) disposed within the purification unit and configured to acquire a pressure value.
4. A high efficiency production apparatus for exosomes according to claim 1, characterised in that: the purification unit comprises:
a first purification mechanism (21) for purifying the input external liquid semi-finished product to obtain an exosome with the diameter less than 200 nm;
a second purification mechanism (22) for purifying the input external liquid semi-finished product to obtain an exosome with the diameter less than 150 nm;
a flow path selection valve (20) for making the external liquid semi-finished product flow into the first purifying mechanism (21) or the second purifying mechanism (22) according to the instruction of the controller (50);
and the material transferring pump (23) is connected with the flow path selection valve (20) and is used for sucking the external liquid semi-finished product to the input end of the flow path selection valve (20).
5. A high efficiency production device for exosomes according to claim 4, characterized in that: the first purifying mechanism (21) comprises:
a first purification tank (211);
a first spray head (212) which is connected with the flow path selection valve (20) and sprays the external liquid semi-finished product into the first purifying box body (211);
a first purification flow rate sensor (216) for acquiring an outflow speed value of exosomes in the first purification box body (211) and sending the outflow speed value to the controller (50);
a first purification carrier (213), said first purification carrier (213) having a cut-off value of 500 kD;
a second purification support (214), the second purification support (214) having a pore size of 200 nm;
a first self-purging control valve (215) for receiving a command of the controller (50) to control the input of the high-pressure gas into the first purification tank (211);
the second purifying mechanism (22) comprises:
a second purification box;
a second nozzle connected with the flow path selection valve (20) to spray the external liquid semi-finished product into the second purifying box;
a second purification flow rate sensor (225) for acquiring an outflow rate value of exosomes in the second purification tank and sending the outflow rate value to the controller (50);
a third purification carrier (223), said third purification carrier (223) having a cut-off value of 500 kD;
a fourth purification support (224), the fourth purification support (224) having a pore size of 200 nm;
a second self-purging control valve (226) for receiving commands from the controller (50) to control the input of high pressure gas into the second purification tank.
6. A high efficiency production device for exosomes according to claim 5, characterized in that: the first purification box body (211) is of an internal hollow structure, a first spray head (212) is installed at the top center of the first purification box body, the distance between the first spray head (212) and a first purification carrier (213) is 2/5 of the whole box body height, the distance between the first purification carrier (213) and a second purification carrier (214) is 1/5 of the whole box body height, the second purification carrier (214) and the first purification carrier (213) are coaxially distributed, the distance between the second purification carrier (214) and the bottom is 1/5 of the whole box body height, a first purification flow rate sensor (216) is arranged at the bottom of the first purification box body (211), the first purification flow rate sensor (216) acquires the current excretion outflow speed and sends the value to the controller (50) in real time, and the controller (50) sends an adjusting instruction to the sample loading pump after processing the data of the first purification flow rate sensor (216) and the value of the detection unit (14) Comparing the acquired data of the first purification flow rate sensor (216) with set parameters, and sending a command to the flow path selection valve (20) or the first self-cleaning control valve (215) when a trigger condition is reached, wherein the first self-cleaning control valve (215) opens the valve after receiving the command, and high-pressure gas sequentially cleans the first purification carrier (213) and the second purification carrier (214) from the upper part; the output end of the first self-cleaning control valve (215) is arranged at the top of the first purification box body (211), a temperature sensor (66) for acquiring the temperature in the box body is arranged on the inner wall of the first purification box body (211), and the temperature sensor (66) sends an acquired value to the controller (50); a pressure sensor (65) for acquiring the tank pressure is arranged at the bottom of the first purification tank body (211), and the pressure sensor (65) sends an acquired value to the controller (50);
the spraying surface of the first spray head (212) is of an arc surface structure, the maximum angle between the arc surface and the horizontal plane is 15 degrees, and the number of spray holes of the spraying surface is 50-80 meshes.
7. A high efficiency production apparatus for exosomes according to claim 2, characterised in that: first chromatography pipe (11) are hollow cylinder structure, and its input and output all are provided with connecting portion (40) that are used for the connecting tube stopper, the opening internal diameter of connecting portion (40) is 1.2-1.5 times of body internal diameter, the wall body of first chromatography pipe (11) is provided with the overhead kick type and is used for the choke ring of spacing compound graphite alkene of carbon nanotube, just choke ring is 2/5 of whole body length apart from the length of input.
8. A high efficiency production device for exosomes according to claim 7, characterized in that: first chromatography pipe (11) are installed on the installation integrated board through chromatography support (30), still install second chromatography pipe (12) and third chromatography pipe (13) through chromatography support (30) on the installation integrated board, just second chromatography pipe (12), third chromatography pipe (13), first chromatography pipe (11) are parallel distribution, and with the horizontal plane of installation integrated board is parallel, the internal diameter of first chromatography pipe (11) is 2 times of second chromatography pipe (12), just the internal diameter of second chromatography pipe (12) is 1.2 times of third chromatography pipe (13) internal diameter, the height homogeneous phase of first chromatography pipe (11), second chromatography pipe (12), third chromatography pipe (13) is the same.
9. A high efficiency production apparatus for exosomes according to claim 8, characterised in that: the input end of the first chromatographic tube (11) is connected with the output end of a sample loading pump (14), the sample loading pump (14) and the pipeline of the first chromatographic tube (11) are provided with a bubble detection component (61), the bubble detection component (61) sends acquired bubble data to a controller (50), the controller (50) sends an execution instruction to the sample loading pump (14), the output end of the first chromatographic tube (11) is provided with a conductance detection component (63) and is connected with the input end of a second chromatographic tube (12), the output end of the second chromatographic tube (12) is provided with a PH detection component (62), the PH detection component (62) sends acquired PH values to the controller (50), a first flow sensor (67) is arranged between the PH detection component (62) and the input end of a third chromatographic tube (13), the output end of the third chromatographic tube (13) is connected with a material transferring pump (23), a UV detection assembly (64) is arranged between the material transferring pump (23) and the third chromatography tube (13), the UV detection assembly (64) sends the wavelength value in the liquid to the controller (50), and the material transferring pump (23) is connected with the flow path selection valve (20);
wherein the output end of the flow path selection valve (20) is provided with a second flow sensor (68), and the second flow sensor (68) sends a pipeline flow speed value to the controller (50).
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| CN202220810800.XU CN217409808U (en) | 2022-04-10 | 2022-04-10 | A high-efficient preparation facilities for exosome |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202220810800.XU CN217409808U (en) | 2022-04-10 | 2022-04-10 | A high-efficient preparation facilities for exosome |
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