An apparatus for large-scale purification and concentration of exosomes

Abstract

The utility model discloses a kind of device of a large number of purified concentrated exosome, belong to exosome purification technical field, including substrate, the top of substrate is fixedly connected with base bar, the outer wall slidingly connected with the clamping structure for positioning to purified concentration structure of base bar, the inner chamber of clamping structure clamps the outer tube for conveying exosome mixed solution flow, the inner chamber of outer tube is fixedly installed with hollow fiber column for exosome purification, the utility model is used, by setting hollow fiber column in the inner chamber of outer tube, and using hollow fiber column to separate exosome and impurity, combined with peristaltic pump stable feed, continuously process a large number of exosome mixed solution, improve the continuity and efficiency of batch purification, satisfy the preparation demand of a large number, and reduce manual intervention, simultaneously through the pressure difference regulation of valve one and valve two, maintain stable purification environment, ensure that exosome is evenly stressed in filtration process, separation is consistent, reduce artificial error, improve product uniformity.

Description

Technical Field

[0001] This invention relates to the field of exosome purification technology, specifically a device for medium-to-large-scale purification and concentration of exosomes. Background Technology

[0002] Exosomes are 30-150nm vesicles secreted by cells that carry bioactive molecules such as proteins and nucleic acids, participate in intercellular communication, and have broad potential for medical applications. They can be used as diagnostic markers, drug carriers, or developed into potential regenerative medicine therapies. They can also serve as efficient drug delivery carriers to achieve targeted delivery of therapeutic components.

[0003] Chinese patent discloses an exosome separation and purification device (authorization announcement number CN222524535U), including a chromatography column and a negative pressure concentration plate. The chromatography column is filled with a packing material with adsorption function. The upper end of the packing material has a space as a sample loading area. The chromatography column has a position for the packing material, which at least partially extends into the ultrafiltration box of the negative pressure concentration plate. The packing material is a chromatographic packing material 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 to the upper end of the bottom box. The bottom box has a box groove. The side of the bottom box is provided with an air extraction port located above the box groove.

[0004] When using the above-mentioned device to purify and separate exosomes, although the device achieves parallel separation and purification of exosomes by setting up multiple chromatography columns, when a large number of exosomes need to be prepared, the adsorption capacity of a single chromatography column is limited, and there are upper limits to the sample loading and separation speed of each chromatography column. A large number of samples need to be processed in batches multiple times, which makes it difficult to achieve continuous processing and results in long processing time. At the same time, when preparing a large number of exosomes, multiple chromatography columns need to be loaded, monitored, and collected simultaneously. The frequency of manual intervention is high, and the separation effect of each chromatography column is easily inconsistent due to differences in operation, which affects the uniformity of the product. Therefore, this utility model provides a device for medium-to-large-scale purification and concentration of exosomes to solve the above problems. Utility Model Content

[0005] The purpose of this invention is to provide a device for purifying and concentrating exosomes in large quantities, so as to solve the problems mentioned in the background art.

[0006] To achieve the above objectives, this utility model provides the following technical solution: A device for medium-volume purification and concentration of exosomes includes a base plate. A base rod is fixedly connected to the top of the base plate. A clamping structure for positioning the purification and concentration structure is slidably connected to the outer wall of the base rod. The inner cavity of the clamping structure holds an outer tube for conveying the flow of an exosome mixture. A hollow fiber column for purifying exosomes is fixedly installed in the inner cavity of the outer tube. A draining structure for diverting waste liquid is fixedly installed on the outer wall of the outer tube. A valve one for testing the exosome injection pressure is fixedly connected to the bottom of the outer tube. A storage structure for providing exosomes to be purified is fixedly connected to the inlet end of the valve one. A valve two for testing the exosome reflux pressure is fixedly connected to the top of the outer tube. The flow rate of exosomes is adjusted by comparing the pressure difference between valve one and valve two to maintain a stable purification and concentration environment. A storage structure for collecting exosomes is fixedly connected to the outlet end of the valve two.

[0007] As a further embodiment of this utility model, the drainage structure includes a drainage pipe, the inlet end of which is fixedly connected to the outer wall of the outer tube, and a storage tank for collecting small molecule impurity waste liquid filtered out by the hollow fiber column is provided below the end of the drainage pipe away from the outer tube.

[0008] As a further embodiment of this utility model, the storage structure includes a conveying pipe, the top of which is fixedly connected to the bottom of valve one, and a peristaltic pump for controlling the delivery rate of the exosomes to be purified is fixedly connected to the outer wall of the conveying pipe. The peristaltic pump stably pushes the liquid to be purified in the storage tank into the outer pipe, and a storage tank for storing the mixture of exosomes to be purified is fixedly connected to the bottom of the peristaltic pump.

[0009] As a further embodiment of this utility model, the storage structure includes a discharge pipe, which is fixedly connected to the top of valve two. A one-way valve two is fixedly connected to the bottom of the outer wall of the discharge pipe to restrict the backflow of purified exosomes. The one-way valve two restricts the one-way flow of the purified and concentrated exosomes. A storage tank for storing the purified exosomes is fixedly connected to the bottom of the one-way valve two, and the purified exosomes are sealed and preserved in the storage tank.

[0010] As a further embodiment of this utility model, the clamping structure includes a support block, which is slidably connected to the outer wall of the base rod. The inner cavity of the support block is threadedly connected to a threaded knob for positioning itself. The inner cavity of the support block is rotatably connected to an arc-shaped secondary rod for clamping the outer tube. The support block and the arc-shaped secondary rod are both fixedly connected to an extension block for clamping the outer wall of the outer tube at the end near the outer tube. The extension blocks are arranged in two groups of four, and the two groups of extension blocks are respectively fixedly connected to the support block and the arc-shaped secondary rod at the end near the outer tube.

[0011] As a further embodiment of this utility model, a conical cup for storing small sterilization tools is fixedly connected to the top of the substrate, and the sterilized small tools are collected and stored in the conical cup.

[0012] Compared with the prior art, the beneficial effects of this utility model are: In use, this invention employs a hollow fiber column within the inner cavity of the outer tube to separate exosomes from impurities. Combined with a peristaltic pump for stable feeding, it continuously processes large-volume exosome mixtures, improving the continuity and efficiency of batch purification, meeting the needs of medium-to-large-volume preparation, and reducing manual intervention. Simultaneously, by regulating the pressure difference between valve one and valve two, a stable purification environment is maintained, ensuring uniform stress and consistent separation of exosomes during filtration, reducing human error, and improving product uniformity. Attached Figure Description

[0013] Figure 1 This is a schematic diagram of the overall structure of a device for purifying and concentrating exosomes in large quantities.

[0014] Figure 2 This is a schematic diagram of the hollow fiber column in a device for purifying and concentrating exosomes in large quantities.

[0015] Figure 3 This is a schematic diagram of the storage tank in a device for purifying and concentrating exosomes in large quantities.

[0016] Figure 4 This is a schematic diagram of the support structure in a device for purifying and concentrating exosomes in large quantities.

[0017] In the diagram: 1. Base plate; 2. Base rod; 3. Clamping structure; 4. Outer tube; 5. Valve 1; 6. Valve 2; 7. Material storage structure; 8. Material storage structure; 9. Drainage structure; 101. Conical cup; 301. Support block; 302. Threaded knob; 303. Arc-shaped secondary rod; 304. Extension block; 305. Push block; 401. Hollow fiber column; 701. Material conveying pipe; 702. Peristaltic pump; 703. Material storage tank; 704. Sealing shell; 705. Extended circular plate; 706. Irregular plate; 707. Hammer block; 708. Motor; 801. Discharge pipe; 802. One-way valve 2; 803. Material storage tank; 901. Drainage pipe; 902. Material storage bucket; 903. One-way valve 1. Detailed Implementation

[0018] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0019] Please see Figures 1-4 In this embodiment of the invention, a device for purifying and concentrating exosomes in large quantities includes a substrate 1. A base rod 2 is fixedly connected to the top of the substrate 1. A clamping structure 3 for positioning the purification and concentration structure is slidably connected to the outer wall of the base rod 2. The inner cavity of the clamping structure 3 holds an outer tube 4 for conveying the flow of the exosome mixture. A hollow fiber column 401 for purifying the exosomes is fixedly installed in the inner cavity of the outer tube 4. A drainage structure 9 for diverting waste liquid is fixedly installed on the outer wall of the outer tube 4. When the exosomes flow upward in the inner cavity of the outer tube 4, the small molecule impurities doped inside permeate to the outside through the membrane pores of the hollow fiber column 401 and are discharged through the drainage structure 9. The exosomes, due to their size, are purified and concentrated. Exosomes larger than the membrane pores are trapped and continue to flow upward with the liquid to complete the purification and concentration. The bottom of the outer tube 4 is fixedly connected to a valve 5 for testing the exosome injection pressure. The outlet end of valve 5 is fixedly connected to the bottom of the outer tube 4. The inlet end of valve 5 is fixedly connected to a storage structure 7 for providing the exosomes to be purified. The top of the outer tube 4 is fixedly connected to a valve 6 for testing the exosome reflux pressure. The inlet end of valve 6 is fixedly connected to the top of the outer tube 4. The flow rate of the exosomes is adjusted by comparing the pressure difference between valve 5 and valve 6 to maintain a stable purification and concentration environment. The outlet end of valve 6 is fixedly connected to a storage structure 8 for collecting the exosomes.

[0020] Please see Figure 1 The drainage structure 9 includes a drainage pipe 901. The inlet end of the drainage pipe 901 is fixedly connected to the outer wall of the outer tube 4. A storage tank 902 is provided below the end of the drainage pipe 901 away from the outer tube 4 to collect the waste liquid containing small molecule impurities filtered by the hollow fiber column 401. Specifically, a one-way valve 903 is fixedly connected to the outer wall of the drainage pipe 901 to restrict the backflow of waste liquid. The one-way valve 903 restricts the one-way flow of waste liquid entering the drainage pipe 901, thereby preventing the waste liquid from contaminating the purified exosomes and ensuring the purity of the product. At the same time, the waste liquid enters the storage tank 902 through the drainage pipe 901 for centralized collection, so that the waste liquid can be treated uniformly after purification. The waste liquid collection process does not require manual intervention, thereby reducing the number of manual dumping or transfer of waste liquid, effectively improving the continuity and efficiency of the overall purification process, and reducing the risk of contamination during operation by reducing the frequency of contact between humans and waste liquid and the device.

[0021] Please see Figure 1 and Figure 3 The storage structure 7 includes a conveying pipe 701. The top of the conveying pipe 701 is fixedly connected to the bottom of valve 5. The outlet end of the conveying pipe 701 is fixedly connected to the inlet end of valve 5. A peristaltic pump 702 for controlling the delivery rate of the exosomes to be purified is fixedly connected to the outer wall of the conveying pipe 701. The peristaltic pump 702 stably pushes the liquid to be purified in the storage tank 703 into the outer pipe 4. The bottom of the peristaltic pump 702 is fixedly connected to the storage tank 703 for storing the mixture of exosomes to be purified. Specifically, a sealing shell 704 is provided at the bottom of the storage tank 703. An extended circular plate 705 for limiting the storage tank 703 is fixedly connected to the top of the sealing shell 704. A shaped plate 706 is rotatably connected to the inner cavity of the sealing shell 704. A hammer block 707 for generating vibration is fixedly connected to the top of the shaped plate 706. Three hammer blocks 707 are provided, and the three hammer blocks 707 are arranged in an axial array. At the top of the irregular plate 706, the tops of the three hammers 707 are all attached to the top of the inner cavity of the sealing shell 704. The bottom of the inner cavity of the sealing shell 704 is fixedly connected to a motor 708 for driving the irregular plate 706 to rotate. The output shaft of the motor 708 is fixedly connected to the bottom of the irregular plate 706. Thus, when the output shaft of the motor 708 drives the irregular plate 706 to rotate, the irregular plate 706 vibrates through friction and impact between the hammers 707 and the top of the inner cavity of the sealing shell 704. This vibration is transmitted through the contact between the sealing shell 704 and the storage tank 703, causing the storage tank 703 to vibrate. This, in turn, causes the mixed liquid to be purified inside to form a slow circulation, preventing the exosomes from separating from the buffer solution, ensuring that the exosomes are evenly distributed in the mixed solution, avoiding fluctuations in the concentration of exosomes entering the outer tube 4 due to sedimentation or aggregation, ensuring the stability of the exosomes during filtration in the hollow fiber column 401, and improving the purification efficiency.

[0022] Please see Figure 1 The storage structure 8 includes a discharge pipe 801, which is fixedly connected to the top of valve 6. The inlet end of the discharge pipe 801 is fixedly connected to the outlet end of valve 6. A one-way valve 802 is fixedly connected to the bottom of the outer wall of the discharge pipe 801 to restrict the backflow of purified exosomes. The one-way valve 802 restricts the unidirectional flow of purified and concentrated exosomes, preventing the purified exosomes from flowing back into the outer pipe 4 and mixing with the unpurified liquid. This avoids contamination or dilution of the purified exosomes and ensures the purity of the purified exosomes. A storage tank 803 is fixedly connected to the bottom of the one-way valve 802 to store the purified exosomes. The storage tank 803 seals and preserves the purified exosomes, reducing structural damage or activity loss caused by exposure to the external environment. It also facilitates centralized collection and subsequent use, providing a stable storage environment for the long-term preservation or immediate application of exosomes.

[0023] Please see Figure 1 and Figure 4 The clamping structure 3 includes a support block 301, which is slidably connected to the outer wall of the base rod 2. The inner cavity of the support block 301 is threadedly connected to a threaded knob 302 for self-positioning. By rotating the threaded knob 302, its end is pressed against the outer wall of the base rod 2, achieving height positioning of the support block 301. The inner cavity of the support block 301 is rotatably connected via a connecting rod to an arc-shaped secondary rod 303 for clamping the outer tube 4. Two sets of arc-shaped secondary rods 303 are provided, and the two sets of arc-shaped secondary rods 303 are rotatably connected to the outer walls of the connecting rods located at both ends of the inner cavity of the support block 301. An extension block 304 for clamping the outer wall of the outer tube 4 is fixedly connected to the end of both the support block 301 and the arc-shaped secondary rod 303 near the outer tube 4. There are two sets of four extension blocks 304, and the two sets of extension blocks 304 are fixedly connected to the support block 301 and the arc-shaped secondary rod 303 at the end near the outer tube 4. Specifically, the ends of the two sets of arc-shaped secondary rods 303 away from the extension blocks 304 are fixedly connected to the levers 305. In use, the outer tube 4 is placed between the four extension blocks 304 on any side of the support block 301, and the levers 305 are turned in sequence to rotate the two arc-shaped secondary rods 303. The two arc-shaped secondary rods 303 drive two of the extension blocks 304 to move closer to the outer wall of the outer tube 4, so that the four extension blocks 304 form a symmetrical clamping and positioning of the outer wall of the outer tube 4. The stability of the outer tube 4 is enhanced by multi-point contact, and it is prevented from shifting due to liquid flow or vibration during the purification process.

[0024] Please see Figure 1 The top of the substrate 1 is fixedly connected to a conical cup 101 for storing small disinfection utensils. The conical cup 101 is used to collect the disinfected small utensils in a centralized manner, so as to facilitate quick retrieval and placement. With its independent space, it can avoid secondary contamination of the utensils after disinfection.

[0025] The working principle of this utility model is as follows: In use, the exosome mixture to be purified is first placed in the storage tank 703, and the storage tank 703 is placed inside the extended circular plate 705. When the output shaft of the drive motor 708 drives the irregular plate 706 to rotate, the irregular plate 706 vibrates through the friction and impact between the hammer block 707 and the top of the inner cavity of the sealing shell 704. The vibration is transmitted through the contact between the sealing shell 704 and the storage tank 703, causing the storage tank 703 to vibrate. This, in turn, causes the mixed liquid to be purified inside to form a slow circulation. Then, the outer tube 4 is placed between the four extension blocks 304 on any side of the support block 301. By turning the lever block 305, the two arc-shaped secondary rods 303 are rotated in sequence, causing the two arc-shaped secondary rods 303 to move two of the extension blocks 304 closer to the outer wall of the outer tube 4. This allows the four extension blocks 304 to symmetrically clamp and position the outer wall of the outer tube 4. Finally, the sterilized small instrument is placed in the base plate. 1. Set up the conical cup 101 for later use and drive the peristaltic pump 702 to introduce the liquid to be purified from the storage tank 703 into the outer tube 4. The mixture flows upward in the outer tube 4. Because the exosomes are larger than the membrane pores of the hollow fiber column 401, they are trapped. As the liquid continues to move upward, small molecule impurities permeate through the membrane pores to the outside of the outer tube 4 and flow into the storage tank 902 through the drain pipe 901. The filtered exosomes enter the storage tank 803 through the discharge pipe 801 for temporary storage. After the exosomes have completely flowed out of the storage tank 703, turn off the peristaltic pump 702 and take out the purified exosomes from the storage tank 803. Add the same volume of PBS buffer as the filtered exosomes to dilute and wash the concentrated exosomes. Pour the mixed exosome solution into another sterilized storage tank 703 and filter again. Repeat this process several times until the permeate is clear and transparent to complete the purification and concentration.

[0026] The above are merely preferred embodiments of this utility model, but the scope of protection of this utility model is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in this utility model, based on the technical solution and inventive concept of this utility model, should be included within the scope of protection of this utility model.

Claims

1. An apparatus for purifying and concentrating exosomes in large quantities, comprising a substrate (1), characterized in that, A base rod (2) is fixedly connected to the top of the substrate (1). A clamping structure (3) for positioning the purification and concentration structure is slidably connected to the outer wall of the base rod (2). An outer tube (4) for conveying the flow of exosome mixture is clamped in the inner cavity of the clamping structure (3). A hollow fiber column (401) for purifying exosomes is fixedly installed in the inner cavity of the outer tube (4). A draining structure (9) for diverting waste liquid is fixedly installed on the outer wall of the outer tube (4). A valve one (5) for testing the exosome injection pressure is fixedly connected to the bottom of the outer tube (4). A storage structure (7) for providing exosomes to be purified is fixedly connected to the inlet end of the valve one (5). A valve two (6) for testing the exosome reflux pressure is fixedly connected to the top of the outer tube (4). The flow rate of exosomes is adjusted by comparing the pressure difference between valve one (5) and valve two (6) to maintain a stable purification and concentration environment. A storage structure (8) for collecting exosomes is fixedly connected to the outlet end of the valve two (6).

2. The apparatus for purifying and concentrating exosomes in large quantities according to claim 1, characterized in that, The drainage structure (9) includes a drainage pipe (901), the inlet end of which is fixedly connected to the outer wall of the outer tube (4), and a storage tank (902) for collecting small molecule impurity waste liquid filtered by the hollow fiber column (401) is provided below the end of the drainage pipe (901) away from the outer tube (4).

3. The apparatus for purifying and concentrating exosomes in large quantities according to claim 1, characterized in that, The storage structure (7) includes a conveying pipe (701), the top of which is fixedly connected to the bottom of valve 1 (5), and a peristaltic pump (702) for controlling the delivery rate of the exosomes to be purified is fixedly connected to the outer wall of the conveying pipe (701), and the liquid to be purified in the storage tank (703) is stably pushed into the outer tube (4) by the peristaltic pump (702), and a storage tank (703) for storing the mixture of exosomes to be purified is fixedly connected to the bottom of the peristaltic pump (702).

4. The apparatus for purifying and concentrating exosomes in large quantities according to claim 1, characterized in that, The storage structure (8) includes a discharge pipe (801), which is fixedly connected to the top of valve two (6). The bottom of the outer wall of the discharge pipe (801) is fixedly connected to a one-way valve two (802) for restricting the backflow of purified exosomes. The one-way valve two (802) restricts the one-way flow of purified and concentrated exosomes. The bottom of the one-way valve two (802) is fixedly connected to a storage tank (803) for storing purified exosomes. The purified exosomes are sealed and preserved by the storage tank (803).

5. The apparatus for purifying and concentrating exosomes in large quantities according to claim 1, characterized in that, The clamping structure (3) includes a support block (301), which is slidably connected to the outer wall of the base rod (2). The inner cavity of the support block (301) is threadedly connected to a threaded knob (302) for positioning itself. The inner cavity of the support block (301) is rotatably connected to an arc-shaped secondary rod (303) for clamping the outer tube (4). The support block (301) and the arc-shaped secondary rod (303) are both fixedly connected to an extension block (304) for clamping the outer wall of the outer tube (4) at one end near the outer tube (4). The extension blocks (304) are arranged in two groups of four, and the two groups of extension blocks (304) are respectively fixedly connected to the support block (301) and the arc-shaped secondary rod (303) at one end near the outer tube (4).

6. The apparatus for purifying and concentrating exosomes in large quantities according to claim 1, characterized in that, The top of the substrate (1) is fixedly connected to a conical cup (101) for storing small sterilized utensils, and the sterilized utensils are collected in a concentrated manner through the conical cup (101).

Images