CN210856074U - High-flux exosome separating device - Google Patents
High-flux exosome separating device Download PDFInfo
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- CN210856074U CN210856074U CN201921236922.7U CN201921236922U CN210856074U CN 210856074 U CN210856074 U CN 210856074U CN 201921236922 U CN201921236922 U CN 201921236922U CN 210856074 U CN210856074 U CN 210856074U
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Abstract
A high-flux exosome separating device comprises a fixed frame and a deep hole plate; the liquid sample comprises exosomes and small molecule substances; the capture magnetic beads comprise CD63 capture magnetic beads and CD9 capture magnetic beads, the CD63 capture magnetic beads comprise exosomes and CD63 transmembrane proteins, the CD9 capture magnetic beads comprise exosomes and CD9 transmembrane proteins, and the exosomes are vesicle structures secreted by cells; the deep hole plate comprises deep holes, 1 reaction tube is arranged in each deep hole, and a liquid sample or capture magnetic beads are filled in the reaction tubes; the deep hole plate is arranged at the middle upper part of the fixing frame; the bottom of the reaction tube is of an ultrafiltration membrane structure, and the ultrafiltration membrane structure is of a cone shape with an opening at the upper end; the bottom end of the deep hole is provided with 1 small hole; the device also comprises a waste liquid collecting tank, a vacuum cavity and an air pump; the waste liquid collecting tank is arranged at the bottom of the fixing frame. The ultrafiltration membrane is a 100KDa ultrafiltration membrane. Compared with the prior art, the beneficial effects of the utility model are that: the utility model has the advantages that: simple structure, scientific and reasonable, simple and convenient operation, high speed and high efficiency, and is suitable for popularization and application.
Description
Technical Field
The utility model discloses a high flux exosome separator, in particular to high flux exosome separator belongs to the biological detection equipment field.
Background
Exosomes (exosomes) were discovered in 1986, and were bilayer vesicular-like structure bodies with a diameter of about 30-100nm, which were actively secreted by various cells in the body, such as immune cells, stem cells, cardiovascular cells, reticulocytes, platelets, nerve cells, and tumor cells, and were widely distributed in body fluids, such as peripheral blood, urine, saliva, milk, ascites, amniotic fluid, etc. The exosome carries a large number of specific proteins (such as cytokines and growth factors) and functional bioactive substances such as mRNAs and miRNAs, participates in physiological processes such as cell communication, cell migration, angiogenesis promotion, anti-tumor immunity and the like in vivo, and is closely related to the occurrence and the process of various diseases. Due to the special structure and function of the exosome, the exosome has potential application value, can be used as a biological index for diagnosing various diseases on one hand, can also be used as a treatment means on the other hand, and can be possibly used as a natural carrier of a medicament for clinical treatment in the future. The separation and purification of exosomes are always the concerns of researchers, and the acquisition of high-purity exosomes is of great importance to the subsequent research. It is known that at present, people mostly adopt methods such as ultracentrifugation, immunomagnetic beads, ultrafiltration, precipitation or kits to realize the extraction and separation of exosomes.
However, in the prior art, the method for separating exosomes is often very complicated, either special complicated instruments and equipment are needed, or a large amount of time is needed, and a high-flux exosome separation device which has a simple structure and can separate exosomes with high efficiency is not available in the prior art.
SUMMERY OF THE UTILITY MODEL
The utility model aims at the above-mentioned prior art in, the separation needs complicated instrument and equipment, and defect such as inefficiency provides a high flux exosome separator, can reach high efficiency, high flux's purpose.
In order to realize the purpose, the utility model adopts the technical scheme that: a high-flux exosome separating device is used for obtaining exosomes through a liquid sample and capture magnetic beads and comprises a fixed frame and a deep hole plate; the liquid sample comprises exosomes and small molecule substances, such as sugar, salt and small molecule protein; the capture magnetic beads comprise CD63 capture magnetic beads and CD9 capture magnetic beads, the CD63 capture magnetic beads comprise exosomes and CD63 transmembrane proteins, the CD63 transmembrane proteins are attached to the surfaces of the exosomes, the CD9 capture magnetic beads comprise exosomes and CD9 transmembrane proteins, and the CD9 transmembrane proteins are attached to the surfaces of the exosomes; the exosome is a vesicle structure secreted by a cell; the deep hole plate comprises deep holes, 1 reaction tube is arranged in each deep hole, and a liquid sample or capture magnetic beads are filled in the reaction tubes; the deep hole plate is arranged at the middle upper part of the fixing frame; the bottom of the reaction tube is of an ultrafiltration membrane structure, and the ultrafiltration membrane structure is of a cone shape with an opening at the upper end; the bottom end of the deep hole is provided with 1 small hole;
the device also comprises a waste liquid collecting tank, a vacuum cavity and an air pump; the waste liquid collecting tank is arranged at the bottom of the fixing frame, the vacuum cavity is arranged in the fixing frame and between the waste liquid collecting tank and the deep hole plate, the whole vacuum cavity is a sealed container, and the upper end of the vacuum cavity is communicated with the inner cavity of the ultrafiltration membrane through a small hole; the air pump is arranged on one side of the vacuum cavity, and one side of the vacuum cavity is connected with the air pump through a pipeline; when the air pump is started, the small molecular substances in the liquid sample can be driven to pass through the ultrafiltration membrane structure and downwards pass through the small hole at the bottom end of the deep hole plate to enter the waste liquid collecting tank;
the one high-throughput exosome separation device is a 96-throughput exosome separation device.
The ultrafiltration membrane fixing device further comprises a clamp (not shown in the figure), the ultrafiltration membrane is fixed by the clamp, the clamp is conventional technology, and details are not repeated.
The ultrafiltration membrane is a 100KDa ultrafiltration membrane.
The exosome has a vesicle structure with the size of 30-150 nm.
The negative pressure in the vacuum cavity is-10 to-30 KPa.
Between the deep holes, a magnetic bar is arranged, which is connected with an automated magnetic bead separation device (not shown in the figure).
Compared with the prior art, the beneficial effects of the utility model are that: the beneficial effects of the utility model reside in that: simple structure, scientific and reasonable, simple and convenient operation, high speed and high efficiency, and is suitable for popularization and application.
Drawings
FIG. 1 is a schematic diagram of: high throughput exosome separation device side view (upper).
FIG. 2 is a diagram of: high throughput exosome separation device side view (lower).
Description of reference numerals: the device comprises a fixed frame 1, a deep-hole plate 2, a deep hole 201, a small hole 202, CD63 captured magnetic beads 3, CD9 captured magnetic beads 4, a reaction tube 5, an ultrafiltration membrane structure 6, a waste liquid collecting tank 7, a vacuum cavity 8 and an air pump 9.
Detailed Description
The present invention will be further described with reference to the accompanying drawings and specific examples, which are not intended to limit the present invention.
As shown in fig. 1-2, a high-throughput exosome separation device for obtaining exosomes from a liquid sample and capture magnetic beads comprises a fixed frame 1 and a deep-well plate 2, wherein the deep-well plate 2 is a 96-well deep-well plate 2; the liquid sample (not shown) comprises exosomes and small molecule substances, such as sugars, salts, small molecule proteins; the capture magnetic beads comprise a CD63 capture magnetic bead 3 and a CD9 capture magnetic bead 4, the CD63 capture magnetic bead 3 comprises an exosome and a CD63 transmembrane protein, the CD63 transmembrane protein is attached to the surface of the exosome, the CD9 capture magnetic bead 4 comprises an exosome and a CD9 transmembrane protein, and the CD9 transmembrane protein is attached to the surface of the exosome; the exosome is a vesicle structure secreted by a cell; the deep hole plate 2 comprises deep holes 201, 1 reaction tube 5 is arranged in each deep hole 201, and a liquid sample or capture magnetic beads are filled in each reaction tube 5; the deep hole plate 2 is arranged at the middle upper part of the fixing frame 1; the bottom of the reaction tube 5 is provided with an ultrafiltration membrane structure 6, and the ultrafiltration membrane structure 6 is in a cone shape with an opening at the upper end; the bottom end of the deep hole 201 is provided with 1 small hole 202;
the device also comprises a waste liquid collecting tank 7, a vacuum cavity 8 and an air pump 9; the waste liquid collecting tank 7 is arranged at the bottom of the fixing frame 1, the vacuum cavity 8 is arranged in the fixing frame 1 and between the waste liquid collecting tank 7 and the deep hole plate 2, the vacuum cavity 8 is integrally a sealed container, and the upper end of the vacuum cavity 8 is communicated with the inner cavity of the ultrafiltration membrane through a small hole 202; the air pump 9 is arranged on one side of the vacuum cavity 8, and one side of the vacuum cavity 8 is connected with the air pump 9 through a pipeline; when the air pump 9 is started, the small molecular substances in the liquid sample can be driven to pass through the ultrafiltration membrane structure 6 and downwards pass through the small hole 202 at the bottom end of the deep hole 201 plate and enter the waste liquid collecting tank 7;
the one high-throughput exosome separation device is a 96-throughput exosome separation device.
The ultrafiltration membrane fixing device further comprises a clamp (not shown in the figure), the ultrafiltration membrane is fixed by the clamp, the clamp is conventional technology, and details are not repeated.
The ultrafiltration membrane is a 100KDa ultrafiltration membrane.
The exosome has a vesicle structure with the size of 30-150 nm.
The negative pressure in the vacuum cavity is-10 to-30 KPa.
Between the deep holes 201, magnetic rods are arranged, which are connected to an automated bead separation device (not shown).
The action mechanism is as follows: the utility model discloses be the vesicle structure of a class of cell secretion according to the exosome, the size is 30-150nm, and CD63 transmembrane protein attaches to the surface of exosome and forms CD63 and catches magnetic bead 3, CD9 transmembrane protein attaches to the surface of exosome and forms these two properties that CD9 caught magnetic bead 4, separates it;
the action process is as follows:
(1) adding a liquid sample into the ultrafiltration membrane structure 6, and starting the air pump 9 to generate negative pressure in the vacuum cavity 8, so that small molecular substances in the sample, such as sugar, salt and small molecular proteins, can enter the waste liquid collecting tank 7 at the bottom of the fixing frame 1 through the ultrafiltration membrane, and the exosomes can be remained in the inner cavity of the ultrafiltration membrane;
(2) closing the air pump 9, adding cleaning solution, standing for 10min, then restarting the air pump 9, adding antigen-antibody binding buffer solution, and enabling exosomes to be combined with the CD63 capture magnetic beads 3 and the CD9 capture magnetic beads 4 to form CD63 capture magnetic beads 3 and CD9 capture magnetic beads 4 respectively;
(3) the separation of exosomes can be realized by starting the automatic magnetic bead separation device, and the separation of exosomes by using the automatic magnetic bead separation device is conventional technology and is not described in detail.
The above-mentioned embodiments are only one of the preferred embodiments of the present invention, and the general changes and substitutions performed by those skilled in the art within the technical scope of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A high-flux exosome separating device is used for obtaining exosomes through a liquid sample and capture magnetic beads and comprises a fixed frame and a deep hole plate; the liquid sample comprises exosomes and small molecule substances; the capture magnetic beads comprise CD63 capture magnetic beads and CD9 capture magnetic beads, the CD63 capture magnetic beads comprise exosomes and CD63 transmembrane proteins, the CD63 transmembrane proteins are attached to the surfaces of the exosomes, the CD9 capture magnetic beads comprise exosomes and CD9 transmembrane proteins, and the CD9 transmembrane proteins are attached to the surfaces of the exosomes; the exosome is a vesicle structure secreted by a cell; the deep hole plate comprises deep holes, 1 reaction tube is arranged in each deep hole, and a liquid sample or capture magnetic beads are filled in the reaction tubes; the deep hole board sets up upper portion in the mount, its characterized in that: the bottom of the reaction tube is of an ultrafiltration membrane structure, and the ultrafiltration membrane structure is of a cone shape with an opening at the upper end; the bottom end of the deep hole is provided with 1 small hole;
the device also comprises a waste liquid collecting tank, a vacuum cavity and an air pump; the waste liquid collecting tank is arranged at the bottom of the fixing frame, the vacuum cavity is arranged in the fixing frame and between the waste liquid collecting tank and the deep hole plate, and the upper end of the vacuum cavity is communicated with the inner cavity of the ultrafiltration membrane through a small hole; the air pump is arranged on one side of the vacuum cavity, and one side of the vacuum cavity is connected with the air pump through a pipeline; when the air pump is started, the micro-molecular substances in the liquid sample can be driven to pass through the ultrafiltration membrane structure and downwards pass through the small hole at the bottom end of the deep hole plate to enter the waste liquid collecting tank.
2. A high throughput exosome separating device according to claim 1, characterized by: the one high-throughput exosome separation device is a 96-throughput exosome separation device.
3. A high throughput exosome separating device according to claim 1, characterized by: the ultrafiltration membrane fixing device further comprises a clamp, and the ultrafiltration membrane is fixed by the clamp.
4. A high throughput exosome separating device according to claim 1, characterized by: the ultrafiltration membrane is a 100KDa ultrafiltration membrane.
5. A high throughput exosome separating device according to claim 1, characterized by: the exosome has a vesicle structure with the size of 30-150 nm.
6. A high throughput exosome separating device according to claim 1, characterized by: the negative pressure in the vacuum cavity is-10 to-30 KPa.
7. A high throughput exosome separating device according to claim 1, characterized by: and a magnetic bar is arranged between the deep holes and is connected with the automatic magnetic bead separation device.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201921236922.7U CN210856074U (en) | 2019-08-01 | 2019-08-01 | High-flux exosome separating device |
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| CN201921236922.7U CN210856074U (en) | 2019-08-01 | 2019-08-01 | High-flux exosome separating device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113005033A (en) * | 2021-02-24 | 2021-06-22 | 清华大学 | Device and method for capturing and separating exosomes and cells in biological sample |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113005033A (en) * | 2021-02-24 | 2021-06-22 | 清华大学 | Device and method for capturing and separating exosomes and cells in biological sample |
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