CN215162612U

CN215162612U - Exosome extraction element

Info

Publication number: CN215162612U
Application number: CN202120795475.XU
Authority: CN
Inventors: 康琳
Original assignee: Peking Union Medical College Hospital Chinese Academy of Medical Sciences
Current assignee: Peking Union Medical College Hospital Chinese Academy of Medical Sciences
Priority date: 2021-04-19
Filing date: 2021-04-19
Publication date: 2021-12-14
Anticipated expiration: 2031-04-19

Abstract

The utility model discloses an exosome extraction element, the device comprises a device body, the top of device body is provided with the introduction port, the bottom is provided with out the appearance mouth, the inside top-down of device body has set gradually filtration membrane, go up the psammitolite, gel filler and lower psammitolite, filtration membrane installs on the inside upper end lateral wall face of device body, go up interval predetermined distance between psammitolite and the filtration membrane, the psammitolite sets up the inside lower extreme at the device body down, form the enclosure space between last psammitolite and the lower psammitolite, it has the gel filler to fill in the enclosure space, be provided with the buffer solution charge door on the outside lateral wall face of device body between filtration membrane and the last psammitolite. The utility model discloses an exosome extraction element can remain exosome vesicle structure and integrality in the at utmost, and the exosome purity that the separation obtained is high, and the disengaging process is simple, and the separation rate is fast, saves time and labour.

Description

Exosome extraction element

Technical Field

The utility model relates to an exosome extraction separation technical field, concretely relates to high-purity integrative exosome extraction element who combines filtration method and gel exclusion chromatography.

Background

Exosomes (exosomes) are small vesicles of about 30-150nm in diameter that can be secreted by most living cells in the body, with a typical lipid bilayer structure, widely found in cell culture supernatants, serum, plasma, saliva, urine, amniotic fluid, cerebrospinal fluid, milk and other biological fluids; exosomes carry various proteins, lipids, RNA and the like, are used as important transfer signal molecules, play an important role in substance and information transfer between cells, and are expected to become early diagnosis markers of various diseases.

At present, the existing exosome extraction and separation equipment mainly separates and extracts exosomes based on methods such as ultracentrifugation, filtration centrifugation, density gradient centrifugation, immunomagnetic beads, a PS affinity method, chromatography and the like. However, these devices still have disadvantages, for example, the purity of exosomes extracted by ultracentrifugation and filtration centrifugation devices is not high; the operation of extracting exosomes by using density gradient centrifugal equipment is complex and time-consuming, and the amount of one-time extraction is small; although expensive instruments are not needed for extracting exosomes by using the immunomagnetic bead method, the non-neutral pH and the non-physiological salt concentration influence the biological activity of exosomes and are not beneficial to carrying out the next experiment; the PS affinity method does not use a denaturant, does not influence the biological activity of exosome, but has slow separation speed and simple operation; the exosome with high purity and structural integrity can be obtained by chromatography extraction, but a complex pretreatment step is required, and integrated exosome extraction cannot be realized.

In summary, in view of the above problems in the prior art, there is a need in the art for a device that is simple in operation, time-saving and labor-saving, and capable of separating exosomes with high purity and complete structure.

SUMMERY OF THE UTILITY MODEL

In view of this, the utility model provides an aim at provides a high-purity integrated exosome extraction element who combines filtration method and gel exclusion chromatography can solve the exosome purity that current equipment extracted not high, the operation is complicated, consuming time, the extraction process influences exosome biological activity scheduling problem.

Based on the above object, an aspect of the embodiment of the utility model provides an exosome extraction element, the device comprises a device body, the top of device body is provided with the introduction port, and the bottom is provided with the appearance mouth, the inside top-down of device body has set gradually filtration membrane, last psammitolite, gel filler and lower psammitolite, filtration membrane installs on the inside upper end lateral wall face of device body, go up the psammitolite with interval predetermined distance between the filtration membrane, the psammitolite sets up down the inside lower extreme of device body go up the psammitolite with form the enclosure down between the psammitolite, the enclosure is filled with the gel filler filtration membrane with go up between the psammitolite be provided with the buffer solution charge door on the outside lateral wall face of device body.

In the exosome extraction device described above, preferably, the device body is a circular cylinder.

In the exosome extraction device as described above, preferably, the filtration membrane is a hydrophilic PVDF membrane.

In the exosome extraction device described above, preferably, the pore size of the hydrophilic PVDF membrane is 0.22 μm.

In the exosome extraction device described above, preferably, the gel filler is a dextran, agarose or polyacrylamide porous material.

In the exosome extraction device, the pore size of the gel filler is preferably 35nm-70 nm.

Advantageous effects

The utility model provides an exosome extraction element is the integrated device who has combined filtration method and gel exclusion chromatography, the device simple structure, low cost, operation labour saving and time saving, inside filtration membrane and the gel filler of having set up of device body, can once only utilize filtration method and gel exclusion chromatography to come the high-purity exosome of quick extraction simultaneously, the separation extraction process is simple, can remain exosome vesicle structure and integrality in the at utmost, the exosome purity that the separation obtained is high, can be used to follow-up treatment and marker detection.

Drawings

Fig. 1 is a schematic diagram of an exosome extraction device according to one embodiment of the present invention.

Detailed Description

The following examples are intended to illustrate the invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

Based on the above-mentioned purpose, the embodiment of the utility model provides an embodiment of exosome extraction element is proposed. Fig. 1 is a schematic diagram of the exosome extraction device of this embodiment. As shown in fig. 1, the exosome-extracting device mainly comprises a pot-shaped housing and an extraction core component inside the housing, thereby constituting the device body of the exosome-extracting device. The top of device body is provided with introduction port 1, and the bottom is provided with outlet 7, and introduction port 1 and outlet 7 can set up between two parties respectively the top and the bottom of device body, the orientation of introduction port 1 are upwards, and the orientation of outlet 7 is downwards. The device body is internally provided with a filtering membrane 2, an upper sand core 4, a gel filler 5 and a lower sand core 6 from top to bottom in sequence. The filtering membrane 2 is installed on the inner upper end side wall surface of the apparatus body, and the upper sand core 4 is spaced from the filtering membrane 2 by a predetermined distance, which may be about half of the height of the apparatus body. The lower sand core 6 is arranged at the lower end of the inner part of the device body, a closed space is formed between the upper sand core 4 and the lower sand core 6, namely the outer edges of the upper sand core 4 and the lower sand core 6 are tightly attached to the inner side wall of the device body. The closed space is filled with the gel filler 5, and the gel filler 5 is formed into a gel column. And a buffer solution feeding port 3 is arranged on the outer side wall surface of the device body between the filtering membrane 2 and the upper sand core 4. The sample inlet 1, the sample outlet 7 and the buffer solution charging hole 3 are communicated with the interior of the device body.

In a preferred embodiment, the device body is a circular cylinder. It is obvious to those skilled in the art that the device body may have other shapes, and those skilled in the art can make the selection as needed as long as the technical problems described above can be solved.

In a preferred embodiment, the filtration membrane is a hydrophilic PVDF membrane having a pore size of 0.22 μm.

In a preferred embodiment, the gel filler is a porous material of dextran, agarose or polyacrylamide, and the pore diameter of the gel filler is 35nm-70 nm.

For those skilled in the art, the above definition of numerical values by the embodiments should not limit the protection scope of the present invention, which is only the preferred embodiments of the present invention. Those skilled in the art can make modifications as needed, as long as they can solve the technical problems described above.

Utilize the utility model discloses a process that exosome extraction element drawed exosome as follows: and adding the solution rich in the exosome into the device body through the sample inlet 1, filtering the solution by using the filtering membrane 2 to remove large-particle impurities such as cell fragments, large vesicles and the like in the solution, and obtaining a filtrate only containing exosome and small-molecule protein. Then, the filtrate flows into a gel exclusion chromatographic column below along the same potential, meanwhile, buffer solution is continuously added through a buffer solution feeding port 3 to wash the gel column, and finally, eluent with fixed volume is collected through a sample outlet 7 to obtain the high-purity exosome enrichment liquid.

To sum up, the utility model provides a combine integrative exosome extraction element of filtration and gel exclusion chromatography, the device simple structure, low cost, operation labour saving and time saving, the inside filtration membrane and the gel filler of having set up of device body, can once only utilize filtration and gel exclusion chromatography to come the high-purity exosome of quick extraction simultaneously, the separation extraction process is simple, can remain exosome vesicle structure and integrality in the at utmost, the exosome purity that the separation obtained is high, can be used to follow-up treatment and marker detection.

Although the invention has been described in detail with respect to the general description and the specific embodiments, it will be apparent to those skilled in the art that modifications and improvements can be made based on the invention. Therefore, such modifications and improvements are intended to be within the scope of the invention as claimed.

Claims

1. The utility model provides an exosome extraction element, a serial communication port, the device comprises a device body, the top of device body is provided with the introduction port, and the bottom is provided with out the appearance mouth, the inside top-down of device body has set gradually filtration membrane, last psammitolite, gel filler and lower psammitolite, filtration membrane installs on the inside upper end lateral wall face of device body, go up the psammitolite with interval predetermined distance between the filtration membrane, the psammitolite sets up down the inside lower extreme of device body go up the psammitolite with form the enclosure down between the psammitolite, it has been filled in the enclosure the gel filler filtration membrane with between the last psammitolite be provided with the buffer solution charge door on the outside lateral wall face of device body.

2. An exosome extraction device according to claim 1, wherein the device body is a circular cylinder.

3. An exosome extraction device according to claim 1, in which the filtration membrane is a hydrophilic PVDF membrane.

4. An exosome extraction device according to claim 3, wherein the pore size of the hydrophilic PVDF membrane is 0.22 μm.

5. An exosome extraction device according to claim 1, in which the gel-packing is a dextran, agarose or polyacrylamide porous material.

6. An exosome extraction device according to claim 5, wherein the pore size of the gel-packing is between 35nm and 70 nm.