CN220696690U - Magnetic bead microsphere preparation device for exosome separation - Google Patents
Magnetic bead microsphere preparation device for exosome separation Download PDFInfo
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- CN220696690U CN220696690U CN202322096799.6U CN202322096799U CN220696690U CN 220696690 U CN220696690 U CN 220696690U CN 202322096799 U CN202322096799 U CN 202322096799U CN 220696690 U CN220696690 U CN 220696690U
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- magnetic bead
- microspheres
- exosome separation
- microsphere
- pressure
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- 239000011324 bead Substances 0.000 title claims abstract description 83
- 239000004005 microsphere Substances 0.000 title claims abstract description 75
- 238000000926 separation method Methods 0.000 title claims abstract description 39
- 210000001808 exosome Anatomy 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 239000000463 material Substances 0.000 claims abstract description 45
- 238000007493 shaping process Methods 0.000 claims abstract description 21
- 238000002156 mixing Methods 0.000 claims abstract description 18
- 238000004140 cleaning Methods 0.000 claims description 9
- 239000012670 alkaline solution Substances 0.000 claims description 8
- 230000006835 compression Effects 0.000 claims description 6
- 238000007906 compression Methods 0.000 claims description 6
- 238000003760 magnetic stirring Methods 0.000 claims description 6
- 238000003860 storage Methods 0.000 claims description 5
- 229920001661 Chitosan Polymers 0.000 abstract description 17
- 239000002245 particle Substances 0.000 abstract description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 18
- 238000000034 method Methods 0.000 description 9
- 239000002994 raw material Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- 239000003570 air Substances 0.000 description 6
- 238000010586 diagram Methods 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 4
- 230000002378 acidificating effect Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002086 nanomaterial Substances 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 108010039918 Polylysine Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000012080 ambient air Substances 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- -1 hydrogen ions Chemical class 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000007885 magnetic separation Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000008558 metabolic pathway by substance Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 238000000710 polymer precipitation Methods 0.000 description 1
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- 238000011084 recovery Methods 0.000 description 1
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- 238000001542 size-exclusion chromatography Methods 0.000 description 1
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- 239000007858 starting material Substances 0.000 description 1
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- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
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Abstract
The utility model discloses a magnetic bead microsphere preparation device for exosome separation, and belongs to the field of exosome separation. The utility model relates to a magnetic bead microsphere preparation device for exosome separation, which comprises a pressure control system, a material mixing system and a magnetic bead microsphere shaping system. The preparation device provided by the utility model realizes semi-automatic and stable operation, can ensure stable quality of the prepared magnetic bead microspheres, and solves the problems of instability, low efficiency and the like existing in manual preparation of chitosan magnetic bead microspheres; meanwhile, the magnetic bead microspheres with different particle sizes can be prepared by adjusting the system pressure provided by a pressure control system and replacing microshunts with different apertures; in addition, the preparation device provided by the utility model has the advantages of simple structure, easiness in disassembly and low field requirement, and is suitable for various occasions.
Description
Technical Field
The utility model relates to a magnetic bead microsphere preparation device for exosome separation, and belongs to the field of exosome separation.
Background
Studies have shown that exosomes play an important role in the metabolism of substances and signaling between cells, and are involved in disease development and immunomodulation. However, efficient separation of exosomes is challenging due to their small size and similar characteristics to other substances in blood. The most commonly used separation method at present is an ultracentrifugation method based on density separation, but the method is long in time consumption, high in cost and low in recovery rate and separation purity. In addition, there are other exosome separation methods such as immunoseparation based on antigen-antibody reaction, ultrafiltration separation based on size and size exclusion chromatography, polymer precipitation separation based on hydrophilic-hydrophobic properties, and microfluidic method depending on fluid mechanics, which have advantages and disadvantages, respectively.
Because exosome membranes carry negative charges, charge-based methods have been increasingly valued in recent years. However, the method of applying an electric field is complicated in operation, and the method using polylysine adsorption is expensive and unsuitable for treating a part of biological samples, so that research on the method of separating exosomes based on charges is still very promising. Chitosan is a natural bioactive compound with amino groups that combine with hydrogen ions under acidic conditions to positively charge chitosan. The chitosan magnetic bead microsphere has the characteristics of high selectivity, high sensitivity, magnetic separation and low injury in exosome separation, can provide an effective and reliable method for purifying and enriching exosomes, and lays a foundation for research and application of exosomes. At present, the preparation of chitosan magnetic bead microspheres lacks special devices, and generally adopts a manual preparation mode, which comprises the processes of solution preparation, stirring and mixing, precipitation and separation, washing, drying and the like.
The chitosan magnetic bead microspheres are manually prepared, each step needs accurate operation and tolerance so as to ensure the preparation quality of the microspheres, and the steps are very complicated. And the quality of the prepared chitosan magnetic bead microspheres may be different due to unavoidable subjective factors of manual operation. This may result in microspheres that vary in size, shape, and magnetic properties, affecting the stability and effectiveness of subsequent applications.
To this end, a magnetic bead microsphere preparation device for exosome separation was developed against the above problems.
Disclosure of Invention
The utility model provides a magnetic bead microsphere preparation device for exosome separation, which aims to solve the problems of complicated steps, different quality, low preparation efficiency and the like in manual preparation of chitosan magnetic beads.
The utility model provides a magnetic bead microsphere preparation device for exosome separation, which comprises a pressure control system, a material mixing system and a magnetic bead microsphere shaping system.
The pressure control system comprises a pressure reducing valve, a flat valve and a pressure input control unit; the pressure reducing valve, the flat valve and the pressure input control unit are sequentially connected through pipelines, one end of the pressure reducing valve is connected with the flat valve through a pipeline, the other end of the pressure reducing valve is externally connected with the compression pump, and the compression pump provides a pressure source.
The material mixing system comprises a material mixer main body; wherein the upper end of the material mixer main body is provided with 3 interfaces; the first interface is an air inlet and is communicated with the pressure control system; the second interface is a deflation port and is connected with a pressure balance switch; the third interface is a discharge port and is communicated with a feeding port of the shaping system through a pipeline extending to the lower end of the material mixer main body.
In one embodiment, the material mixing device body further comprises a magnetic stirring rod, and the magnetic stirring rod is positioned at the bottom of the material mixing device body.
In one embodiment, all 3 interfaces in the material mixing system are connected with the outside through hoses.
In one embodiment, the material mixer comprises a raw material for preparing magnetic bead microspheres for exosome separation; the starting materials are routinely selected by the person skilled in the art in view of the prior art. For example, the raw materials are chitosan and Fe as disclosed in example 2 of CN116376124A 3 O 4 A mixed solution of particles, or a mixed solution of Fe/chitosan in example 1 of patent document CN108607517B, or the like.
In one embodiment, the material mixer is chitosan/Fe 3 O 4 A granular liquid mixer.
The magnetic bead microsphere shaping system comprises a shaping system feed inlet, a micro-diverter connected with the tail end of the feed inlet, and a magnetic bead microsphere shaper positioned at the lower end of the micro-diverter.
In one embodiment, the microsyringe has a pore diameter of less than 0.7mm.
In one embodiment, the microsyringe is in the form of a needle having a diameter of less than 0.7mm.
In one embodiment, the magnetic bead microsphere shaping system further comprises a magnetic bead rapid cleaning hair receiving plate. The device can be used as a magnetic bead microsphere cleaning device, and the magnetic beads in alkaline solution are cleaned and then dried for use.
In one embodiment, the magnetic bead microsphere shaper is an alkaline solution tank, such as a sodium hydroxide solution tank, alternatively a tank containing 3M NaOH solution as disclosed in example 2 of CN116376124 a.
The utility model has the advantages that:
(1) According to the magnetic bead microsphere preparation device for exosome separation, the pressure is provided by the pressure control system, so that the materials of the material mixing device automatically enter the micro-flow divider to be extruded, and enter the magnetic bead microsphere shaping device to be shaped to obtain the magnetic bead microsphere; the device realizes semi-automatic and stable operation, can ensure stable quality of the prepared magnetic bead microspheres, and solves the problems of instability, low efficiency and the like in the manual preparation of the chitosan magnetic bead microspheres.
(2) The magnetic bead microsphere preparation device for exosome separation can be used for preparing magnetic bead microspheres with different particle sizes, and can be realized by adjusting the system pressure provided by a pressure control system and replacing microshunts with different apertures. The exosome separation magnetic bead microsphere preparation device can be used for preparing customized magnetic bead microspheres.
(3) The device has the advantages of easy acquisition of components required by construction, low cost, simple structure, easy disassembly and the like.
(4) The magnetic bead microsphere preparation device for exosome separation has low requirements on sites, is suitable for various occasions, can be used in school scientific research institutions, small laboratories and can also be used for industrial production by amplifying systems.
Drawings
FIG. 1 is a schematic diagram of a magnetic bead microsphere preparation apparatus for exosome separation;
FIG. 2 is a schematic diagram of a material blending system;
FIG. 3 is a schematic diagram of a magnetic bead microsphere shaping system;
wherein: 1 is a pressure reducing valve; 2 is a flat valve; 3 is a pressure input control unit; 4 is a pressure balance switch; 5 is a material mixer main body; 6 is a magnetic stirring rod; 7 is a micropipette; 8 is a magnetic bead microsphere shaper; 9 is a magnetic bead rapid cleaning hair receiving disc; 10 is an air inlet; 11 is a discharge hole; 12 is taken as a raw material; 13 is a magnetic stirrer; 14 is a feeding port of the shaping system; 15 is an alkaline solution.
Detailed Description
The present utility model will be described in detail below.
Example 1
FIG. 1 is a schematic diagram of a magnetic bead microsphere preparation apparatus for exosome separation according to the present utility model.
The utility model relates to a magnetic bead microsphere preparation device for exosome separation, which comprises a pressure control system, a material mixing system and a magnetic bead microsphere shaping system.
The pressure control system comprises a pressure reducing valve 1, a flat valve 2 and a pressure input control unit 3; the pressure reducing valve 1, the flat valve 2 and the pressure input control unit 3 are sequentially connected through pipelines, one end of the pressure reducing valve 1 is connected with the flat valve 2 through a pipeline, the other end of the pressure reducing valve is externally connected with the compression pump, and the compression pump provides a pressure source.
As shown in fig. 2, a schematic diagram of the material mixing system includes a material mixer body 5; wherein the upper end of the material mixer main body is provided with 3 interfaces; the first interface is an air inlet 10 which is communicated with a pressure control system; the second interface is a deflation port and is connected with the pressure balance switch 4; the third interface is a discharge port 11 which is communicated with a feeding port of the shaping system through a pipeline extending to the lower end of the material mixer main body 5.
Fig. 3 shows a schematic diagram of the magnetic bead microsphere shaping system, which includes a shaping system feed inlet 14, a micro-diverter 7 connected to the tail end of the feed inlet, and a magnetic bead microsphere shaper 8 located at the lower end of the micro-diverter 7.
The material mixer main body also comprises a magnetic stirring rod 6, is positioned at the bottom of the material mixer main body 5 and is used together with a magnetic stirrer 13 which is externally arranged below the material mixer main body 5.
The material mixer comprises a raw material 12 for preparing exosome separation magnetic bead microspheres; those skilled in the art can routinely choose from the prior art. Polymers (which may be chitosan) and magnetic nanomaterials (which may be magnetic Fe) mentioned for preparing polymer modified magnetic nanomaterials as disclosed in CN 115554992A 3 O 4 Nanoparticles), again such as the chitosan and Fe involved as disclosed in example 2 of CN116376124a 3 O 4 The mixed solution of particles is also, for example, the Fe/chitosan mixed solution in example 1 of patent document CN 108607517B.
The micro-shunt 7 is a needle with a diameter smaller than 0.7mm.
The magnetic bead microsphere shaper 8 is a sodium hydroxide solution storage tank, and can be a storage tank containing 3M NaOH solution disclosed in the example 2 of CN 116376124A.
Optionally, the magnetic bead microsphere shaping system further comprises a magnetic bead rapid cleaning and hair receiving disc 9.
Optionally, all 3 interfaces in the material mixing system are connected with the outside through hoses.
The working principle of the utility model is as follows:
the pressure reducing valve 1, the flat valve 2, the pressure input control unit 3, the material mixer system and the micro-dispenser 7 are sequentially arranged into a closed and communicated space according to the figure 1, wherein the pressure reducing valve 1, the flat valve 2, the pressure input control unit 3 and the pressure balance switch 4 are air pressure passages, and power is provided by an external pressure supply system to control the pressure of the whole closed space; the material mixer body 5 is internally provided with a raw material for preparing magnetic bead microspheres, and can be routinely selected by a person skilled in the art according to the prior art; the magnetic stirring rod 6 can be used together with a magnetic stirrer 13 arranged outside below the material mixer main body 5 and is used for mixing materials in the material mixer main body 5; the aperture of the micro-shunt 7 is smaller than 0.7mm, and the micro-shunt is used for separating magnetic bead microspheres; the bead shaper 8 is filled with a certain amount of an alkaline solution, such as NaOH solution, which can be routinely chosen by the person skilled in the art according to prior art. Based on the characteristics of chitosan dissolution under acidic conditions and precipitation under alkaline conditions, the beads dripped in the microporous shunt 7 can form magnetic bead microspheres with good integrity and uniform size in the magnetic bead microsphere shaper 8 (NaOH solution storage tank); the magnetic bead rapid cleaning and receiving disc 9 can be used as a magnetic bead microsphere cleaning device, and the magnetic beads in alkaline solution are cleaned and dried for use.
The working principle and working process of the device of the utility model are as follows:
(1) Before preparing exosome separation magnetic bead microspheres: the raw material 12 for preparing chitosan magnetic bead microspheres is added into the material mixer body 5, 3 ports are connected in the above manner, then the magnetic stirrer 13 is turned on to stir the raw material, and then the pressure supply system is turned on: firstly, the power supply is switched on, the tightness of the positive pressure flow pipeline and the material mixing system is checked (the air release valve 4 needs to be firstly adjusted to the maximum gear), the compression pump switch is turned on, and the pressure switch is adjusted to be zero. The technician adjusts the pressure provided by the pressure control system according to the actual needs.
(2) When preparing exosome separation magnetic bead microspheres: in the material mixer, as shown in fig. 2, the aerodynamic direction is from the air inlet to the discharge outlet, so as to achieve a certain balance. Raw materials in the material mixer 5 flow out from a discharge hole 11 under the action of system pressure, and drop into a magnetic bead microsphere shaper 8 through a shaping system feed hole 14 and a micro-diverter 7 connected with the tail end of the feed hole. The pressure control system can control the speed of the liquid drops flowing out of the micro-diverter 7, thereby affecting the preparation efficiency of the magnetic bead microspheres. Meanwhile, the microshunt 7 with different apertures can be replaced, and the size of the magnetic bead microsphere can be controlled by combining the pressure provided by the pressure control system. The magnetic bead microsphere shaper 8 is an alkaline solution storage tank, can play a role in shaping magnetic bead microspheres, and needs to pay attention to that the alkaline solution needs to be overflowed from the microspheres. In addition, the magnetic rod 6 at the inner bottom of the material mixer body 5 combines with the external magnetic stirrer 13 to complete the material mixing action, and simultaneously, the material mixer body 5 is fixed.
(3) After preparing exosome separation magnetic bead microspheres: the pressure balance switch 4 is connected to balance the ambient air pressure, so that potential safety hazards are avoided, and then the pressure supply system is closed, so that the device stops working. And cleaning and drying the prepared magnetic bead microspheres through a magnetic bead rapid cleaning and hair receiving disc 9.
While the utility model has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the utility model as defined in the appended claims.
Claims (8)
1. The magnetic bead microsphere preparation device for exosome separation is characterized by comprising a pressure control system, a material mixing system and a magnetic bead microsphere shaping system;
the material mixing system comprises a material mixer main body; wherein the upper end of the material mixer main body is provided with 3 interfaces; the first interface is an air inlet and is communicated with the pressure control system; the second interface is a deflation port and is connected with a pressure balance switch; the third interface is a discharge port and is communicated with a feed port of the shaping system through a pipeline extending to the lower end of the material mixer main body;
the magnetic bead microsphere shaping system comprises a shaping system feed inlet, a micro-diverter connected with the tail end of the feed inlet, and a magnetic bead microsphere shaper positioned at the lower end of the micro-diverter.
2. The apparatus for preparing magnetic bead microspheres for exosome separation according to claim 1, wherein the pressure control system comprises a pressure reducing valve, a flat valve, and a pressure input control unit; the pressure reducing valve, the flat valve and the pressure input control unit are sequentially connected through pipelines, one end of the pressure reducing valve is connected with the flat valve through a pipeline, and the other end of the pressure reducing valve is externally connected with the compression pump.
3. The apparatus for preparing magnetic bead microspheres for exosome separation according to claim 1, wherein the material mixer body further comprises a magnetic stirring rod positioned at the bottom inside the material mixer body.
4. The device for preparing magnetic bead microspheres for exosome separation according to claim 1, wherein the 3 interfaces in the material mixing system are all connected with the outside through hoses.
5. The apparatus for preparing magnetic bead microspheres for exosome separation according to claim 1, wherein the aperture of the micro-diverter is less than 0.7mm.
6. The apparatus for preparing magnetic bead microspheres for exosome separation according to claim 1, wherein the micro-diverter is in the form of a needle with a diameter of less than 0.7mm.
7. The device for preparing magnetic bead microspheres for exosome separation according to claim 1, wherein the magnetic bead microsphere shaping system further comprises a magnetic bead rapid cleaning and hair receiving disc.
8. The apparatus for preparing magnetic bead microspheres for exosome separation according to claim 1, wherein the magnetic bead microsphere shaper is an alkaline solution storage tank.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322096799.6U CN220696690U (en) | 2023-08-03 | 2023-08-03 | Magnetic bead microsphere preparation device for exosome separation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322096799.6U CN220696690U (en) | 2023-08-03 | 2023-08-03 | Magnetic bead microsphere preparation device for exosome separation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN220696690U true CN220696690U (en) | 2024-04-02 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202322096799.6U Active CN220696690U (en) | 2023-08-03 | 2023-08-03 | Magnetic bead microsphere preparation device for exosome separation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN220696690U (en) |
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2023
- 2023-08-03 CN CN202322096799.6U patent/CN220696690U/en active Active
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