Exosome separation and purification device

Abstract

The utility model provides an exosome separation and purification device and relates to the technical field of medical instruments. Comprising a chromatographic column and a negative pressure concentration plate, the chromatographic column is internally provided with filler with an adsorption function, and the upper end of the filler is provided with a space serving as a sample adding area; the chromatographic column is provided with a filler position, and at least part of the chromatographic column extends into the ultrafiltration box of the negative pressure concentration plate; wherein the filler is a chromatographic filler with a protein adsorption function or an exosome adsorption function; the negative pressure concentration plate is also provided with a bottom box, the side surface of the ultrafiltration box is hermetically connected to the upper end of the bottom box, and a box groove is formed in the bottom box; and the side surface of the bottom box is provided with an extraction opening above the box groove. The exosome separation and purification device combines the advantages of a chromatographic column and a negative pressure concentration plate, has the characteristics of multifunctionality, simplicity and convenience in operation, less sample loss, good sealing performance and the like, and is suitable for separation and purification processes of exosomes.

Description

Technical Field

[0001] The utility model relates to the technical field of medical devices, and in particular to an exosome separation and purification device. Background Art

[0002] Exosomes are small vesicles released by cells, which contain various biomolecules produced in cells, such as proteins, RNA, DNA, lipids, etc. Exosomes can transmit information and regulate cell activities by interacting with other cells or tissues. This intercellular communication mechanism is widely present in organisms and plays an important role in various physiological and disease processes, such as immune response, cell signaling, tumor formation and progression, etc. The study of exosomes is of great significance for a deeper understanding of cell communication, disease mechanisms, and the development of new treatments.

[0003] In the process of studying exosomes, it is often necessary to separate and purify exosomes, but the current separation equipment is too complicated. For example, in Chinese patent No. 202122021701.1, an exosome separation device is disclosed, which is not only complex in structure, but also easy to lose cell activity by centrifugal separation. Therefore, the utility model proposes an exosome separation and purification device to at least partially solve the problems that may exist in the prior art. Utility Model Content

[0004] The purpose of the utility model is to provide an exosome separation and purification device, which can propose solutions to the deficiencies of the prior art, combines the advantages of a chromatography column and a negative pressure concentration plate, has the characteristics of versatility, simple operation, less sample loss, good sealing, etc., and is suitable for the separation and purification process of exosomes.

[0005] In order to solve the above technical problems, the technical solution adopted by the utility model is:

[0006] An exosome separation and purification device comprises a chromatography column and a negative pressure concentration plate; the chromatography column is provided with a filler with an adsorption function, and the upper end of the filler has a space as a sample addition area; the chromatography column has a position for the filler, which at least partially extends into an ultrafiltration box of the negative pressure concentration plate; wherein the filler is a chromatographic filler with a protein or exosome adsorption function; the negative pressure concentration plate is also provided with a bottom box, the side of the ultrafiltration box is sealed and connected to the upper end of the bottom box, and a box groove is provided in the bottom box; an air suction port is provided on the side of the bottom box above the box groove.

[0007] Furthermore, in the utility model, the chromatography column and the negative pressure concentration plate can be separately arranged.

[0008] Furthermore, in the utility model, a filter outlet is also provided at the lower end of the chromatography column. When the chromatography column is connected to the negative pressure concentration plate, the filter outlet is at least partially located in the ultrafiltration box.

[0009] Furthermore, in the present invention, the portion of the ultrafiltration box located inside the bottom box is at least partially made of ultrafiltration material.

[0010] Furthermore, in the present invention, the chromatography column is a 96-well chromatography column, wherein the 96-well chromatography column includes a single tube arrangement and / or an 8-tube arrangement.

[0011] Furthermore, in the utility model, the air suction port protrudes from the bottom box and is made of hard colloid.

[0012] The present application also discloses an exosome separation and purification device, comprising:

[0013] A tubular container, in which a sample addition area, a filler and an ultrafiltration box are arranged from top to bottom; the filler is a chromatographic filler with the function of adsorbing proteins or exosomes;

[0014] A bottom box, which has a box groove inside and an air extraction port on the side of the bottom box located at the box groove;

[0015] The tubular container and the bottom box ensure that at least the portion of the ultrafiltration box where the ultrafiltration material is located is located inside the bottom box.

[0016] Furthermore, in the present invention, the tubular containers are arranged in an array in number of 96.

[0017] Furthermore, in the present invention, the ultrafiltration material is a nano-ultrafiltration membrane.

[0018] The present application also discloses an exosome separation and purification device, comprising: a chromatography column and a negative pressure concentration plate;

[0019] The chromatography column is tubular and is arranged on the negative pressure concentrator; the negative pressure concentrator has an ultrafiltration component inside, a box slot is provided at the lower end of the ultrafiltration component, and an air extraction port is provided outside the ultrafiltration component for extracting air to form a negative pressure inside;

[0020] The chromatography column has a filler disposed therein and at least one opening disposed at the upper end thereof, wherein the filler is a chromatographic filler having the function of adsorbing proteins or adsorbing exosomes.

[0021] Furthermore, in the utility model, the chromatography column is provided with a piston in its nozzle.

[0022] or,

[0023] The chromatography column is provided with two pipe openings, one of which is provided with a piston, and the other pipe opening is provided with a pipe plug.

[0024] The utility model has at least the following advantages or beneficial effects:

[0025] Through the chromatography column and the negative pressure concentration plate; the chromatography column is provided with a filler with an adsorption function, and the upper end of the filler has a space as a sample addition area; the chromatography column has a position for the filler, which at least partially extends into the ultrafiltration box of the pressure concentration plate; wherein the filler is a chromatographic filler with the function of adsorbing proteins or exosomes; the negative pressure concentration plate is also provided with a bottom box, the side of the ultrafiltration box is sealed and connected to the upper end of the bottom box, and a box slot is provided in the bottom box; the side of the bottom box is provided with an air extraction port at a position above the box slot. The negative pressure concentration plate and the chromatography column can form a sealed connection, and the speed of chromatography can be accelerated while the negative pressure is applied. The exosome separation and purification device combines the advantages of the chromatography column and the negative pressure concentration plate, and has the characteristics of versatility, easy operation, less sample loss, good sealing, etc. It is suitable for the separation and purification of exosomes in batches and without damage, ensuring the integrity, high dispersion and high biological activity of the exosomes. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solutions of the embodiments of the utility model, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the utility model and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying creative work.

[0027] Figure 1 A schematic diagram of the structure of an exosome separation and purification device provided in one embodiment of the utility model;

[0028] Figure 2 A schematic diagram of the working process of an exosome separation and purification device provided in one embodiment of the utility model;

[0029] Figure 3 A schematic diagram of the structure of a three-dimensional perspective of an exosome separation and purification device provided in one embodiment of the utility model;

[0030] Figure 4 A schematic diagram of the structure of various types of chromatography columns of an exosome separation and purification device provided in one embodiment of the utility model.

[0031] In the attached drawings: 1, chromatography column; 2, negative pressure concentration plate; 11, sample addition area; 12, filler; 13, filter outlet; 21, ultrafiltration box; 22, bottom box; 23, air extraction port; 24, box slot. DETAILED DESCRIPTION

[0032] In order to make the objectives, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] Example 1

[0034] Please refer to Figures 1 to 3 As shown in the figure, this embodiment provides an exosome separation and purification device, including: a chromatography column 1 and a negative pressure concentration plate 2; the chromatography column 1 is provided with a filler 12 with an adsorption function, and the upper end of the filler 12 has a space as a sample addition area 11; the chromatography column 1 has a position for the filler 12, which at least partially extends into the ultrafiltration box 21 of the negative pressure concentration plate 2, wherein the filler 12 is a chromatographic filler with a protein adsorption function; the negative pressure concentration plate 2 is also provided with a bottom box 22, the side of the ultrafiltration box 21 is sealed and connected to the upper end of the bottom box 22, and the bottom box 22 is provided with a box groove 24; the side of the bottom box 22 is provided with an air suction port 23 at a position above the box groove 24.

[0035] In this embodiment, the above-mentioned exosome separation and purification device solves the problem of complex structure and design of current separation and purification devices, simplifies the design of the separation and purification device, and saves the device cost; the device combines the functions of the chromatography column and the negative pressure concentration plate, can simultaneously separate and concentrate exosomes, and reduces the operation steps and equipment costs; through the filler adsorption function, the above-mentioned chromatography column 1 is provided with a filler 12 with the function of adsorbing proteins or exosomes. In this application, the filler 12 with the function of adsorbing proteins is preferred, so as to realize the separation of proteins in the mixture and achieve the purification of exosomes. The position of the filler 12 in the above-mentioned chromatography column 1 extends to the ultrafiltration box 21 of the pressure concentration plate 2, so that the separated exosomes can directly enter the ultrafiltration box for concentration treatment, avoiding the secondary transmission and loss of the exosomes; the side of the ultrafiltration box 21 is sealed with the upper end of the bottom box 22 to ensure that there will be no gas leakage during the concentration process, ensure the suction force at the bottom, and improve the safety and stability of the operation; the side of the above-mentioned bottom box 22 is provided with an exhaust port 23, and the negative pressure of the ultrafiltration plate can be adjusted by exhausting air to achieve better concentration effect and flow control.

[0036] In addition, when different fillers for adsorbing exosomes are used with adsorbed proteins, the negative pressure concentration plate 2 can be replaced by a waste liquid collection plate. A filler 12 with the function of adsorbing exosomes can also be set in the above-mentioned chromatography column 1, and the required exosomes are adsorbed to the above-mentioned filler 12, and the exosomes are selectively adsorbed to remove protein impurities in the sample. At this time, a waste liquid collection plate can be placed under the chromatography column 1 for waste liquid collection (not shown in the figure), and the impurity liquid is collected and discarded by setting the chromatography column 1, and after adding the eluent to the chromatography column 1, the exosomes are eluted into the ultrafiltration box 21 for ultrafiltration concentration.

[0037] It should be noted that the application scope of the above-mentioned exosome separation and purification device is not only in the purification and extraction of exosomes, but also includes virus purification, blood product pretreatment and other directions, which makes it have a wider range of application scenarios and market promotion value.

[0038] Example 2

[0039] This embodiment provides an exosome separation and purification device, wherein, referring to Figure 1 As shown, the chromatography column 1 and the negative pressure concentrator plate 2 can be separately arranged, so that the chromatography column 1 and the negative pressure concentrator plate 2 can be operated independently, which is convenient for different experimental needs. You can choose to use only one part or use both parts at the same time as needed, which improves the flexibility and diversity of operation.

[0040] As a preferred embodiment, the lower end of the chromatography column 1 is also provided with a filter outlet 13. When the chromatography column 1 and the negative pressure concentration plate 2 are connected, the filter outlet 13 is at least partially located in the ultrafiltration box 21. By virtue of the filter outlet 13 at the lower end of the chromatography column 1 being located in the ultrafiltration box 21, the substance separated from the chromatography column 1 can directly enter the ultrafiltration box 21 for further processing, thereby reducing the transmission process of the substance, avoiding the loss and contamination of the substance, and improving the purification efficiency and the purity of the sample.

[0041] As a preferred embodiment, the bottom and / or side of the ultrafiltration box 21 located in the bottom box 22 is made of ultrafiltration material, wherein the bottom and / or side of the ultrafiltration box 21 located in the bottom box 22 is made of ultrafiltration material, and the ultrafiltration material is a nano-ultrafiltration membrane, which can achieve selective separation of molecular size; the ultrafiltration material can exclude larger molecules by screening molecular size, thereby better purifying the target substance.

[0042] As an example, the above-mentioned nano-ultrafiltration membrane is preferably a hydrophilic nanofiltration membrane, which can be specifically a polycarbonate membrane, a nylon membrane, and a glass fiber membrane; in addition, the above-mentioned ultrafiltration material can be one of the following; among them, polyamide membrane (Polyamide membrane) is a polymer synthetic membrane with high separation performance and chemical stability, and is usually used in biotechnology, drinking water treatment, medicine and other fields; polysulfide membrane (Polysulfone membrane) is a material with good chemical resistance and heat resistance, and is usually used in seawater desalination, industrial wastewater treatment and food processing and other fields; polyethersulfone membrane (Polyethersulfone membrane) has good chemical resistance and high temperature stability, and is often used in medicine, electronics and chemical industries. Field); polyamide ether membrane (Polyamide ether membrane) has excellent separation performance and chemical stability, and is often used in bioengineering and water treatment and other fields.

[0043] As a preferred embodiment, the chromatography column 1 is a 96-well chromatography column 1, wherein the 96-well chromatography column 1 includes a single tube arrangement and / or an 8-tube arrangement, referring to Figure 3 As shown, the chromatography column 1 is a 96-well chromatography column 1; the chromatography column 1 is designed as a 96-well chromatography column 1, which can process multiple groups of samples at the same time, improving the high throughput and efficiency of sample analysis. The purified exosomes can be directly eluted into a 96-well plate, which is convenient for high-throughput downstream applications.

[0044] As a preferred implementation method, refer to Figure 3 As shown, the air suction port 23 protrudes from the bottom box 22 and is made of a hard colloid; wherein the air suction port 23 made of a hard colloid has higher pressure resistance and sealing performance, can better control the pressure and flow in the bottom box 22, and realize a more stable and precise negative pressure concentration process.

[0045] Example 3

[0046] The present embodiment provides an exosome separation and purification device, which includes: a tubular container, in which a sample addition area 11, a filler 12 and an ultrafiltration box 21 are arranged from top to bottom; the filler 12 is a chromatographic filler with a protein adsorption function; a bottom box 22, in which a box groove 24 is arranged, and a suction port 23 is arranged on the side of the box groove 24, which is used to evacuate the gas in the bottom box to form a vacuum therein, thereby accelerating the filtration speed during ultrafiltration; the box groove 24 is arranged in the bottom box, which can be used to store liquid after ultrafiltration, and the upper suction port 23 is horizontally located on the box groove 24, so that the liquid in the box groove 24 will not leak from the suction port when the gas is pumped; the tubular container and the bottom box 22 are such that at least the part of the ultrafiltration box 21 provided with the ultrafiltration material is located in the bottom box 22.

[0047] The exosome separation and purification device of the present invention has a working process such as Figure 2 As shown, the sample is first added to the sample adding area 11 of the device, and then the sample is initially adsorbed and filtered through the chromatographic filler with protein adsorption function. Multiple filtrations can also be formed through the above-mentioned multiple sample adding areas 11 and fillers 12. The solution after preliminary filtration and purification is ultrafiltered and separated by the ultrafiltration box 21 below, and the separated and purified liquid is collected by the box slot 24 above. It can also be made into a multi-level chromatography column connected from top to bottom, and the middle is made sealed, so that the overall speed is faster when the negative pressure is drawn below.

[0048] Furthermore, the tubular containers are arranged in an array with 96 pieces, which can simultaneously process 96 groups of exosomes to be separated and purified, thereby improving the processing efficiency.

[0049] Example 4

[0050] This embodiment provides an exosome separation and purification device, referring to Figure 1 As shown, it includes: a chromatography column 1, which is tubular and has a filler 12 therein, and at least one opening is provided at the upper end thereof; when the filler 12 is a chromatographic filler having the function of adsorbing proteins but not adsorbing exosomes, a negative pressure concentration plate 2 is provided at the lower end of the chromatography column 1; the negative pressure concentration plate 2 has an ultrafiltration component therein, and a box slot 24 is provided at the lower end of the ultrafiltration component, and an air suction port 23 is provided outside the negative pressure concentration plate 2 for pumping air to form a negative pressure therein. The negative pressure concentration plate 2 is pumped through the air suction port 23 to form a negative pressure (the internal pressure is less than the external pressure), which is more efficient during filtration. The tubular chromatography column 1 makes it easy for users to add samples to be processed to the device, and the operation is simple. The ultrafiltration box 21 is located in the inner area of ​​the bottom box 22, and is at least partially made of ultrafiltration material.

[0051] As a preferred embodiment, when the filler 12 is a chromatographic filler that adsorbs exosomes but does not adsorb free proteins, a waste liquid collection plate is provided at the lower end of the chromatography column 1. The negative pressure concentration plate 2 is replaced by the waste liquid collection plate, and the waste liquid containing free proteins is collected by the waste liquid collection plate because the exosomes are adsorbed instead of the free proteins. After the process is completed, the lower end of the above-mentioned column can be connected to the negative pressure concentration plate 2 again, and the exosomes are eluted to the negative pressure concentration plate 2 by the eluent for purification and filtration; during the intermediate elution process, the adsorbed exosomes in the chromatography column 1 can also be eluted to other containers, such as a measuring cup, and then transferred to the negative pressure concentration plate 2, or transferred to a vessel with ultrafiltration function for ultrafiltration purification.

[0052] As a preferred implementation method, refer to Figure 4 The chromatography column 1 is also provided with a piston 14 in its nozzle.

[0053] As a preferred implementation method, refer to Figure 4 As shown, the chromatography column 1 is provided with two pipe openings, one of which is provided with a piston 14 and the other pipe opening is provided with a pipe plug 15.

[0054] In this embodiment, the piston 14 is used to perform reciprocating adsorption, repeatedly aspirating and pumping liquid to achieve the best effect of removing impurities. The chromatographic column 1 and the concentrator plate 2 are separated, so that the number of cycles of back-absorption ranges from 10 to 20 times; the filler volume can reach 1 mL / hole, thereby improving the utilization rate of the chromatographic column 1 and the purity after filtration.

[0055] It should be noted that the above-mentioned repeated suction method requires adding a collection plate to the above-mentioned chromatography column 1 and the negative pressure concentration plate 2 so that it can be adsorbed repeatedly, thereby improving the purity.

[0056] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may be subject to various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

[0057] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0058] Although the preferred embodiments of the utility model embodiments have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the utility model embodiments.

[0059] Finally, it should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or terminal device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or terminal device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or terminal device including the elements.

[0060] The above is a detailed introduction to an exosome separation and purification device provided by the utility model. In this article, specific examples are used to illustrate the principle and implementation method of the utility model. The description of the above embodiments is only used to help understand the method and core idea of ​​the utility model; at the same time, for those skilled in the art, according to the idea of ​​the utility model, there will be changes in the specific implementation method and application scope. In summary, the content of this specification should not be understood as a limitation on the utility model.

Claims

1. An exosome separation and purification device, characterized in that: include: Chromatographic column (1) and negative pressure concentration plate (2); The chromatography column (1) is provided with a filler (12) having an adsorption function, and the upper end of the filler (12) has a space serving as a sample addition area (11); The chromatography column (1) has a position for a filler (12), which at least partially extends into the ultrafiltration box (21) of the negative pressure concentration plate (2); wherein the filler (12) is a chromatographic filler having the function of adsorbing proteins or exosomes; The negative pressure concentrator plate (2) is also provided with a bottom box (22), the side of the ultrafiltration box (21) is sealed and connected to the upper end of the bottom box (22), and a box groove (24) is provided in the bottom box (22); and an air suction port (23) is provided on the side of the bottom box (22) at a position above the box groove (24).

2. The exosome separation and purification device according to claim 1, characterized in that: The chromatography column (1) and the negative pressure concentration plate (2) can be arranged separately.

3. The exosome separation and purification device according to claim 1 or 2, characterized in that: The lower end of the chromatography column (1) is also provided with a filter outlet (13). When the chromatography column (1) is connected to the negative pressure concentration plate (2), the filter outlet (13) is at least partially located in the ultrafiltration box (21).

4. The exosome separation and purification device according to claim 1 or 2, characterized in that: The portion of the ultrafiltration box (21) located inside the bottom box (22) is at least partially made of ultrafiltration material.

5. The exosome separation and purification device according to claim 1, characterized in that: The chromatography column (1) is a 96-well chromatography column (1), wherein the 96-well chromatography column (1) comprises a single tube arrangement and / or an 8-tube arrangement.

6. The exosome separation and purification device according to claim 1 or 2, characterized in that: The air suction port (23) protrudes from the bottom box (22) and is made of hard colloid.

7. An exosome separation and purification device, characterized in that: include: A tubular container, in which a sample addition area (11), a filler (12) and an ultrafiltration box (21) are arranged from top to bottom; the filler (12) is a chromatographic filler that has the function of adsorbing proteins but not adsorbing exosomes; or the filler (12) is a chromatographic filler that adsorbs exosomes but does not adsorb free proteins; A bottom box (22) is provided with a box groove (24) inside, and a suction port (23) is provided on the side of the bottom box (22) located at the box groove (24); The tubular container and the bottom box (22) enable at least the portion of the ultrafiltration box (21) where the ultrafiltration material is located to be located inside the bottom box (22).

8. The exosome separation and purification device according to claim 7, characterized in that: The ultrafiltration material is a nano-ultrafiltration membrane.

9. An exosome separation and purification device, characterized in that: include: A chromatography column (1) is tubular and has a filler (12) disposed therein and at least one opening disposed at its upper end; When the filler (12) is a chromatographic filler that has the function of adsorbing proteins but does not adsorb exosomes, a negative pressure concentration plate (2) is provided at the lower end of the chromatography column (1); an ultrafiltration component is provided inside the negative pressure concentration plate (2), and a box groove (24) is provided at the lower end of the ultrafiltration component, and an air extraction port (23) is provided outside the ultrafiltration component for extracting air to form a negative pressure inside; or, When the filler (12) is a chromatographic filler that adsorbs exosomes but does not adsorb free proteins, a waste liquid collection plate is provided at the lower end of the chromatography column (1).

10. The exosome separation and purification device according to claim 9, characterized in that: The chromatography column (1) is also provided with a piston (14) in its nozzle. or, The chromatography column (1) is provided with two pipe openings, one of which is provided with a piston (14) and the other pipe opening is provided with a pipe plug (15).

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