An exosome purification device

Abstract

The application provides an exosome purification device, and relates to the technical field of exosome purification, and comprises a shell, a liquid inlet and outlet interface, a flushing interface and a negative pressure interface which are arranged on the shell and are communicated with the inner cavity of the shell; a basic filter part which comprises a first flow guide and a first filter membrane; and a new filter part which comprises a second flow guide and a second filter membrane, wherein the second flow guide has a predetermined distance from the first flow guide, a sealing channel is arranged between the first flow guide and the second flow guide, and the first filter membrane is sleeved on the outer periphery of the sealing channel. Through the above arrangement, the problem of local blockage of the filter membrane in the purification process is solved, cross contamination is effectively prevented, the purification efficiency is improved, the purification time of 1L samples is shortened to less than five hours, the purification efficiency is improved by 40%, and the exosome recovery rate of the sample to be purified reaches more than 80%.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of exosome purification, in particular to an exosome purification device. BACKGROUND

[0002] As an important intercellular communication medium, exosomes have wide application value in the fields of disease diagnosis, drug delivery and biological research. Exosome purification is a key step to obtain high-purity exosomes, and the purification efficiency and the quality of the purified exosomes directly affect the reliability of the subsequent experimental results.

[0003] At present, there are few exosome purification products on the market for large volume (more than 1L) biological sample exosome purification, and the operation and purification time is too long. It usually takes 15-20 hours to complete the exosome purification of 1L sample, and some even longer. There are fewer exosome purification products with a single loading capacity of more than 10L, and the existing filter membrane-based purification products still have the following problems: first, the effective filtration area of the single-layer filter membrane structure is limited, and it takes a long time to process large volume samples, and the purification efficiency is low; second, manual intervention or additional equipment is required for exosome recovery during the elution stage, which is time-consuming and labor-intensive, and is prone to cause damage to the exosomes, resulting in a low qualified rate of the recovered exosomes; third, the filter membrane is prone to clogging during long-term operation, resulting in a loss of effective filtration area and a decrease in filtration efficiency, which may even affect the stability of continuous operation of the device; in addition, if the filtered waste liquid is not discharged in time or mixed with the sample to be purified, cross contamination may occur. Therefore, it is of great practical significance to develop a new type of exosome purification device. SUMMARY

[0004] The present application provides an exosome purification device to overcome the deficiencies of the prior art.

[0005] The technical solution of the present application to solve the above technical problems is as follows: an exosome purification device, comprising: a shell, the shell is provided with a liquid inlet and outlet interface, a flushing interface and a negative pressure interface which communicate with the inner cavity of the shell;

[0006] A basic filtration part, which includes a first flow guide arranged in the inner cavity of the shell, and a first filter membrane covering the side of the first flow guide away from the inner side wall of the shell, the first flow guide is configured to allow the filtered liquid to pass through, the first filter membrane and the first flow guide are both provided with through holes in the center, and the area between the first flow guide and the inner side wall of the shell is configured as a negative pressure liquid discharge cavity, and the negative pressure interface is connected to the negative pressure liquid discharge cavity;

[0007] A new filter part is added, which includes a second flow guide arranged on the side of the first filter membrane away from the first flow guide, and a second filter membrane covering the side of the second flow guide away from the first filter membrane. The second flow guide has a predetermined distance from the first flow guide, and is configured to prevent the filtered liquid from passing through. The second flow guide has a through hole in the center. A sealed channel for conveying the filtered liquid of the new filter part is arranged between the first flow guide and the second flow guide. One end of the sealed channel is connected to the through hole in the center of the first flow guide, and the other end is connected to the through hole in the center of the second flow guide. The first filter membrane is sleeved on the outer periphery of the sealed channel.

[0008] Further, a support boss for supporting the basic filter part is arranged on the side of the negative pressure drainage cavity.

[0009] A drainage plate is further arranged in the negative pressure drainage cavity. The drainage plate has a honeycomb-shaped through hole penetrating through its upper and lower surfaces. The drainage plate has a certain distance from the first flow guide.

[0010] Further, the first flow guide and the second flow guide have the same diameter. The cross-sectional shape of the sealed channel is circular, and the diameter D1 is 0.02-0.4 times the diameter of the first flow guide or the second flow guide.

[0011] Further, the vertical distance L3 between the second flow guide and the first flow guide is 0.02-1.5 times the diameter D1 of the sealed channel.

[0012] Further, the first flow guide is edge-wrapped. The side of the first flow guide abutting against the first filter membrane is provided with a plurality of arrayed flow guide ribs. The other side of the first flow guide close to the negative pressure drainage cavity is provided with a plurality of arrayed reinforcing plates abutting against the flow guide ribs. The spacing between adjacent flow guide ribs is the same as the spacing between adjacent reinforcing plates, and the length extension directions of the flow guide ribs and the reinforcing plates are perpendicular to each other. The flow guide ribs and the reinforcing plates are staggered to form a channel for the filtered liquid to pass through.

[0013] Further, the width L2 of the reinforcing plate is 2.3-3.5 times the spacing between adjacent flow guide ribs or the spacing L1 between adjacent reinforcing plates.

[0014] Further, the second flow guide is a plate structure with a central hole. The side of the second flow guide abutting against the second filter membrane is a flow guide side. The outer peripheral edge of the flow guide side is edge-wrapped, and a plurality of linear protrusions are uniformly arranged around the center. The protrusions include long ribs and short ribs arranged at intervals. One end of the long rib extends to the edge of the central hole, and the other end extends to the edge of the central hole. One end of the short rib extends to the edge of the central hole, and the other end extends to a predetermined distance from the central hole. The included angle α between adjacent linear protrusions is 5-15 degrees.

[0015] Further, the relationship between the distance K between the short edge end and the center through hole and the included angle α between the adjacent straight convex edges satisfies: wherein the unit of K is centimeter.

[0016] Further, the second flow guide is away from the second filter membrane side is a drainage side, the drainage side is provided with a plurality of arc convex edges uniformly distributed around the center, one end of the arc convex edge extends to the outer peripheral edge of the drainage side, the other end extends to a predetermined distance from the center through hole, the area S1 enclosed by the connecting line between the curvature centers of the plurality of arc convex edges is 0.3-0.7 times the area of the drainage side.

[0017] Further, the shell includes a left shell and a right shell, the left shell and the right shell are both provided with a basic filter part and a new filter part, the new filter part in the left shell is oppositely arranged with the new filter part in the right shell and is spaced apart by a certain distance, and the liquid inlet and outlet interface is arranged at the lower part of the shell and is communicated to the area between the two new filter parts.

[0018] The left shell and the right shell are both provided with a negative pressure interface, and the two negative pressure interfaces are both arranged at the lower part of the shell.

[0019] The beneficial effects of the present application are:

[0020] ①By arranging the double-layer filter unit without interference, and by negative pressure guiding purification, the filtration area is doubled without increasing the radial size of the purification device, the purification efficiency is effectively improved, it is suitable for large volume sample purification, the sample purification time of 1L sample is shortened to less than five hours, the purification efficiency is improved by 40%, and the exosome recovery rate of the purified sample reaches more than 80%.

[0021] ②By arranging the flushing interface, the cleaning liquid forms a high flow rate and high frequency switching cyclone, which can make the target exosomes adsorbed on the surface of the filter membrane fall off, realize the automatic elution and recovery of the exosomes, effectively save the labor cost, improve the work efficiency, and the residual rate of the target exosomes trapped on the filter membrane is less than 10%.

[0022] ③During the purification process, by controlling the high frequency pulse change of the negative pressure, the filter membrane is slightly vibrated and agitated, which can make the target exosomes adsorbed on the surface of the filter membrane loose, solve the problem of local blockage of the filter membrane during the purification process, further ensure the purification efficiency, so that the blockage rate of the purification device is less than 8% during 24 hours continuous operation, and the effective filtration area loss rate is less than or equal to 10%.

[0023] ④By bonding and sealing the filter membrane edge and the corresponding flow guide edge, the waste liquid is directly discharged into the negative pressure liquid discharge cavity, which can avoid mixing of the waste liquid and the sample to be purified, and effectively prevent cross contamination. BRIEF DESCRIPTION OF DRAWINGS

[0024] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the specific embodiments or prior art description will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor on the basis of these drawings.

[0025] Figure 1 A three-dimensional structural schematic diagram of the exosome purification device provided in Embodiment 1 of the present application is shown in the figure.

[0026] Figure 2 A front view of the exosome purification device provided in Embodiment 1 of the present application is shown in the figure.

[0027] Figure 3 A Figure 2 cross-sectional view along A-A direction.

[0028] Figure 4 A Figure 3 enlarged view of A.

[0029] Figure 5 A three-dimensional structural schematic diagram of the liquid discharge plate provided in Embodiment 1 of the present application is shown in the figure.

[0030] Figure 6 A three-dimensional structural schematic diagram of one side of the flow guide rib of the first flow guide provided in Embodiment 1 of the present application is shown in the figure.

[0031] Figure 7 A three-dimensional structural schematic diagram of one side of the reinforcing plate of the first flow guide provided in Embodiment 1 of the present application is shown in the figure.

[0032] Figure 8 A front view of the flow guide side of the second flow guide provided in Embodiment 1 of the present application is shown in the figure.

[0033] Figure 9 A front view of the drainage side of the second flow guide provided in Embodiment 1 of the present application is shown in the figure.

[0034] The drawings show: 1, inlet and outlet interface; 2, flushing interface; 3, negative pressure interface; 4, first flow guide; 41, flow guide rib; 42, reinforcing plate; 5, first filter membrane; 6, negative pressure liquid discharge cavity; 7, second flow guide; 71, straight convex rib; 711, long rib; 712, short rib; 72, arc convex rib; 8, second filter membrane; 9, sealing channel; 10, support boss; 11, liquid discharge plate; 12, shell. DETAILED DESCRIPTION

[0035] The technical solutions of the present application will be described clearly and completely below in connection with the embodiments. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of protection of the present application.

[0036] To make the purposes, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions of the embodiments of the present application will be described clearly and completely below in connection with the drawings of the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of protection of the present application. Therefore, the following detailed description of the embodiments of the present application provided in the drawings is not intended to limit the scope of the claimed present application, but only represents selected embodiments of the present application.

[0037] In the present application, unless specifically defined and limited otherwise, the terms "mounting", "connection", "connecting", "fixed", and the like should be interpreted broadly, for example, can be fixed connection, can also be detachable connection, or integral; can be mechanical connection, can also be electrical connection; can be directly connected, can also be indirectly connected through an intermediate medium, can be the internal communication of two elements or the interaction relationship between two elements. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances. In addition, the terms "first", "second", "third" and the like are only used to distinguish the description, and cannot be understood as indicating or implying relative importance.

[0038] In the present application, unless specifically defined and limited otherwise, the first feature above or below the second feature can include that the first and second features are in direct contact, or that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, the first feature above, above and above the second feature includes that the first feature is directly above and obliquely above the second feature, or only indicates that the horizontal height of the first feature is higher than that of the second feature. The first feature below, below and below the second feature includes that the first feature is directly below and obliquely below the second feature, or only indicates that the horizontal height of the first feature is less than that of the second feature.

[0039] Example 1

[0040] As Figures 1-4As shown, the present application provides an exosome purification device, comprising: a shell 12, which is provided with a liquid inlet and outlet interface 1, a flushing interface 2 and a negative pressure interface 3 communicating with the inner cavity of the shell, the liquid inlet and outlet interface 1 is used for inputting the sample to be purified or outputting the eluted exosome mixed liquid; the flushing interface 2 is connected with an external pipeline, and the external pipeline is connected with a peristaltic pump or other power source, so that the cleaning liquid forms a high flow rate and high flow rate switching cyclone to elute and recover the target exosome filtered in the inner cavity of the shell 12, and the cleaning liquid adopts a known type in the exosome purification field, which will not be described here; the negative pressure interface 3 is connected with an external drainage pipeline, and the external drainage pipeline is connected with a negative pressure generating device, such as a vacuum pump, which is used to establish a negative pressure environment, guide the rapid filtration of the sample to be purified and timely discharge the waste liquid after filtration, preferably, during the purification process, high-frequency pulse type negative pressure change is generated by controlling the start and stop of the negative pressure generating device;

[0041] The basic filtration part comprises a first flow guide 4 arranged in the inner cavity of the shell 12, and a first filter membrane 5 covering the side of the first flow guide 4 away from the inner side wall of the shell, the first flow guide 4 is configured to allow the filtered liquid to pass through, the area of the first filter membrane 5 is not less than the area of the first flow guide 4, and both have a through hole in the center, the area between the first flow guide 4 and the inner side wall of the shell 12 is configured as a negative pressure drainage cavity 6, and the negative pressure interface 3 communicates with the negative pressure drainage cavity 6; it can be understood that the first flow guide 4 is located at the inlet position of the negative pressure drainage cavity 6, under the guidance of negative pressure, the sample to be purified passes through the first filter membrane 5 and the first flow guide 4 in turn, the target exosome is intercepted on the surface of the first filter membrane 5 during the flow process, and the filtered waste liquid enters the negative pressure drainage cavity 6 through the first flow guide 4 and is discharged in time through the negative pressure interface 3, which can avoid the problem of raw material waste caused by discharging the sample to be purified without purification;

[0042] The filter unit further comprises a second flow guide 7 arranged on the side of the first filter membrane 5 away from the first flow guide 4, and a second filter membrane 8 arranged on the side of the second flow guide 7 away from the first filter membrane 5, the second flow guide 7 is arranged at a predetermined distance from the first flow guide 4, and the second flow guide 7 is configured to prevent the filtered liquid from passing through, the area of the second filter membrane 8 is not less than the area of the second flow guide 7, the second flow guide 7 is provided with a through hole in the center, and the first flow guide 4 and the second flow guide 7 are provided with a sealed channel 9 for conveying the filtered liquid of the new filter unit, one end of the sealed channel 9 is connected to the through hole in the center of the first flow guide 4, and the other end is connected to the through hole in the center of the second flow guide 7, and the first filter membrane 5 is sleeved on the outer periphery of the sealed channel 9. It can be understood that the sealed channel 9 leads the negative pressure to the second filter membrane 8, and when the shell 12 is filled with the sample to be purified, the sample to be purified is purified by the basic filter unit, and at the same time, under the guidance of negative pressure, the target exosome is intercepted on the surface of the second filter membrane 8, and the filtered waste liquid enters the second flow guide 7 and flows to the negative pressure liquid discharge cavity 6 through the sealed channel 9; It should be noted that the edge of the first filter membrane 5 is adhesively fixed to the edge of the first flow guide 4, and the edge of the second filter membrane 8 is adhesively fixed to the edge of the second flow guide 7, so that the probability of re-mixing of the filtered waste liquid into the sample to be purified is further reduced, and the sealed channel 9 is connected and fixed between the first flow guide 4 and the second flow guide 7 by heat welding or adhesion.

[0043] Preferably, according to the actual production needs, N new filter units can be arranged, and the center of the second filter membrane of each new filter unit is provided with a through hole, so as to further increase the filtering area. It can be understood that, in order to ensure the smooth discharge of the waste liquid of each new filter unit, the new filter unit and the new filter unit can be connected by a sealed channel, and the connection between the sealed channel and the second flow guide is provided with a gap for waste liquid flow, or the center through hole of the second filter membrane of the new filter unit and the center through hole of the second flow guide of the upper new filter unit can be connected by a heat sealing channel.

[0044] Firstly, by setting double-layer filter units that do not interfere with each other and by negative pressure guiding purification, the filtering area is doubled without increasing the radial size of the purification device, effectively improving the purification efficiency, suitable for large-volume sample purification, shortening the purification time of 1L sample to less than five hours, improving the purification efficiency by 40%, and the recovery rate of exosomes in the purified sample reaching more than 80%; Secondly, by setting a washing interface, the cleaning liquid forms a high-flow-rate cyclone with high-frequency flow rate switching, which can make the target exosomes adsorbed on the surface of the filter membrane fall off when the target exosomes are eluted and recovered, realizing the automatic elution and recovery of exosomes, effectively saving labor costs and improving work efficiency, and the residual rate of target exosomes trapped on the filter membrane is less than 10%; Thirdly, because the filter membrane has a certain flexibility, during the purification process, by controlling the high-frequency pulse change of the negative pressure, the filter membrane is slightly vibrated and inflated, which can make the target exosomes adsorbed on the surface of the filter membrane loose, solve the problem of local blockage of the filter membrane during the purification process, further ensure the purification efficiency, so that the blockage rate of the purification device is less than 8% when it is continuously operated for 24 hours, and the effective filtering area loss rate is less than or equal to 10%; Finally, by bonding and sealing the edges of the filter membrane and the corresponding flow guide, the waste liquid is directly discharged into the negative pressure liquid discharge chamber, which can avoid the mixing of waste liquid and sample to be purified, effectively eliminating the cross contamination problem.

[0045] Specifically, as Figures 3-5As shown, the negative pressure drainage cavity 6 is provided with a support boss 10 on the side for supporting the basic filter part, and the first flow guide 4 abuts against the support boss 10 on the side edge. The negative pressure drainage cavity 6 is further provided with a drainage plate 11, which is provided with honeycomb-shaped through holes penetrating the upper and lower surfaces thereof, and the drainage plate 11 is a certain distance away from the first flow guide 4. It should be noted that the support boss 10 and the shell 12 can be an integral structure, or can be fixed by welding, gluing, clamping and the like, which will not be described here. The first flow guide 4 is fixed to the support boss 10 by heat welding or adhesion. By providing the drainage plate 11, the technical effects achieved are various. First, under the guidance of high negative pressure, the disordered turbulent flow of the purified waste liquid entering the negative pressure drainage cavity 6 at a high flow rate can be effectively dispersed by the drainage plate 11, so as to solve the problem that the kinetic energy of the fluid directly impacts the bottom of the first flow guide 4 under high negative pressure, causing the first flow guide 4 to vibrate and even deform and crack. Secondly, by providing the honeycomb-shaped through holes in the drainage plate 11, the structural strength of the drainage plate 11 is ensured, and a multidirectional flow channel is formed by the hexagonal array structure, so as to fully slow down the waste liquid and reduce the turbulent intensity of the waste liquid, so that the waste liquid remains in a laminar flow state when the flow rate is greater than 1 m / s. Finally, by providing a certain distance between the drainage plate 11 and the first flow guide 4, the resonance problem of the drainage plate 11 and the first flow guide 4 caused by the turbulent flow and pulse negative pressure of the waste liquid can be avoided, a buffer layer can be formed to absorb the negative pressure pulse fluctuation, the reverse backflow of the waste liquid to the bottom of the first flow guide 4 and the first filter membrane 5 caused by the pulse negative pressure can be avoided, the cross-contamination problem caused by the backflow of the waste liquid can be solved, and the problem of the particulate matter in the waste liquid blocking the first flow guide 4 can be solved.

[0046] Specifically, as Figure 4As shown, the first flow guide 4 and the second flow guide 7 have the same diameter, and the sealing channel 9 has a circular cross-sectional shape with a diameter D1 of 0.02-0.4 times the diameter of the first flow guide 4 or the second flow guide 7. First, by setting the first flow guide 4 and the second flow guide 7 to have the same diameter, the structural symmetry can be ensured, the alignment assembly of the basic filtration part and the new filtration part is facilitated, the stress concentration coefficient of the connection between the two ends of the sealing channel 9 and the first flow guide 4 or the second flow guide 7 in the working process is reduced, and the structural stability is ensured. Second, if the cross-sectional diameter D1 of the sealing channel 9 is less than 0.02 times the diameter of the first flow guide 4 or the second flow guide 7, the flow resistance of the waste liquid will increase sharply due to the too small diameter of the sealing channel 9, thereby reducing the flow rate and affecting the filtration efficiency. In addition, the negative pressure transmitted to the new filtration part is insufficient to act on the edge area of the second filter membrane 8, resulting in a loss of 20% of the effective filtration area of the second filter membrane 8. Moreover, the small diameter of the sealing channel 9 leads to a small structural strength, which is easily damaged by fatigue cracking under external stress during the purification process. If the cross-sectional diameter D1 of the sealing channel 9 is greater than 0.4 times the diameter of the first flow guide 4 or the second flow guide 7, too much flow guide space will be occupied, resulting in a small effective support area of the second flow guide 7 for the second filter membrane 8. In a high negative pressure working environment, the center area of the second filter membrane 8 is easily pulled and collapsed, thereby causing some sealing channels 9 to be blocked, affecting the waste liquid discharge of the new filtration part. In addition, a large-diameter through hole needs to be provided in the center of the second flow guide 7, which weakens the structural strength of the second flow guide 7 and causes it to deform under the action of negative pressure, thereby shortening the service life. Therefore, by limiting the diameter D1 of the sealing channel 9 to 0.02-0.4 times the diameter of the flow guide, the service life of the purification device can be prolonged while ensuring high flow rate and high flux.

[0047] Specifically, as Figure 4As shown, the vertical distance L3 between the second flow guide 7 and the first flow guide 4 is 0.02-1.5 times the diameter D1 of the sealing channel 9. When performing elution and recovery of target exosomes, because the sealing channel 9 occupies the central region, its physical structure will block the movement path of the rotational flow of the cleaning liquid, destroy the rotational flow state of the cleaning liquid, and cause the exosome residual rate in the central region of the first filter membrane 5 to be too high. By limiting the vertical distance L3 between the second flow guide 7 and the first flow guide 4 to be 0.02-1.5 times the diameter D1 of the sealing channel 9, after the cleaning liquid is injected through the flushing interface 2, the space between the second flow guide 7 and the first flow guide 4 is compressed due to the change in space, so that part of the rotational flow kinetic energy of the cleaning liquid is converted into turbulent flow kinetic energy, thereby ensuring that the surface of the first filter membrane 5 can be completely covered by the cleaning liquid, avoiding excessive diffusion of the rotational flow kinetic energy of the cleaning liquid due to the excessively large spacing between the first flow guide 4 and the second flow guide 7, causing the flow rate in the central region of the rotational flow to decay, and reducing the elution efficiency. Through this limitation, the exosome residual rate on the first filter membrane 5 is <5% after elution and recovery; and by limiting the vertical distance between the second flow guide 7 and the first flow guide 4, the exosome residual rate on the first filter membrane 5 is reduced, while avoiding the sealing channel 9 being too long, thereby ensuring the structural strength and improving the working stability, and reducing the negative pressure differential between the basic filtration part and the newly added filtration part.

[0048] Preferably, the vertical distance L3 between the second flow guide 7 and the first flow guide 4 is 1.1-1.4 times the diameter D1 of the sealing channel 9. Although the transmission of negative pressure to the newly added filtration part through the sealing channel 9 can improve the filtration efficiency, in the process of the sealing channel 9 collecting and transporting the waste liquid of the newly added filtration part, the radial flow of the waste liquid changes rapidly to axial flow in the sealing channel 9, which causes the flow rate in the center of the sealing channel 9 to be greater than the flow rate at the edge of the channel, causing boundary layer separation, forming a low-pressure vortex zone inside the sealing channel 9, and causing cell fragments and protein aggregates in the waste liquid to be retained in the vortex zone and crosslink into clumps, blocking the sealing channel 9 and reducing the cross-sectional area of the channel, increasing the negative pressure drop loss, and further reducing the waste liquid transport efficiency and the filtration flux of the second filter membrane 8. By limiting the vertical distance L3 between the second flow guide 7 and the first flow guide 4 to be 1.1-1.4 times the diameter D1 of the sealing channel 9, the length of the sealing channel 9 is determined, so that the waste liquid has a sufficient length of liquid flow pattern recovery zone, ensuring that the waste liquid can adhere to the inner wall of the sealing channel 9 during downward flow, ensuring stable laminar flow of the waste liquid in the sealing channel 9, inhibiting boundary layer separation, and avoiding blockage of the sealing channel 9. Under the premise of ensuring the reduction of the exosome residual rate on the first filter membrane 5 and the reduction of the negative pressure differential, the waste liquid transport efficiency and the filtration flux of the second filter membrane 8 are further ensured.

[0049] Specifically, as shown in FIG. 1, the second flow guide 7 is a hollow cylinder, and the first flow guide 4 is a hollow cylinder. Figures 6-7As shown, the first flow guide 4 is edge-wrapped, and the side of the first flow guide 4 abutting the first filter membrane 5 is provided with a plurality of arrayed flow guide ribs 41, and the other side close to the negative pressure liquid drainage cavity 6 is provided with a plurality of arrayed reinforcing plates 42 abutting the flow guide ribs 41, the spacing between adjacent flow guide ribs 41 is the same as the spacing between adjacent reinforcing plates 42, and the length extension directions of the two are perpendicular to each other, and the flow guide ribs 41 and the reinforcing plates 42 are staggered to form channels for the filtered liquid to pass through. It can be understood that the edge-wrapped treatment refers to that the first flow guide 4 is provided with a planar area at the periphery for bonding and fixing with the first filter membrane 5, and under the condition of high negative pressure use, the flow guide ribs 41 disperse the partial load of the first filter membrane 5 to multiple points of support, effectively reducing the loss rate of the filtering area of the first filter membrane 5, and the reinforcing plates 42 form a rigid grid skeleton, which can not only improve the bending strength of the first flow guide 4, but also cooperate with the flow guide ribs 41 to reduce the filtering area loss rate of the first filter membrane 5 while blocking and guiding the negative pressure direction, effectively reducing the deformation amount of the first filter membrane 5 under the influence of negative pressure; secondly, by setting the spacing between the flow guide ribs 41 and the reinforcing plates 42 to be the same and the length extension directions of the two to be perpendicular to each other, the first flow guide 4 forms a regular waste liquid flow channel, the flow guide ribs 41 guide the waste liquid to flow quickly in the vertical direction, and the reinforcing plates 42 block the horizontal turbulent flow and suppress the turbulent intensity of the waste liquid flow, forcing the waste liquid to flow in one direction, reducing the problem of impurity particle flow, avoiding the problems of cross contamination caused by waste liquid backflow and impurity particle blockage of the back of the first filter membrane 5, and this setting can ensure uniform distribution of support points, uniform distribution of stress, effectively prolong the service life, and also avoid the problem of impurity deposition caused by sudden drop of flow velocity of waste liquid in wide channels due to different spacings, so that the blockage rate of the purification device in 24-hour continuous operation is less than 6%.

[0050] Specifically, as Figures 6-7As shown, the width L2 of the reinforcing plate 42 is 2.3-3.5 times the spacing between adjacent guide ribs 41 or the spacing between adjacent reinforcing plates 42. First, through this arrangement, the relationship between the support performance of the first guide member 4 and the stability of the waste liquid flow can be further balanced, while ensuring that the first guide member 4 has sufficient structural strength, the problem of excessive flux attenuation of the first filter membrane 5 caused by the increase in flow resistance and the decrease in waste liquid flow rate due to the excessive width of the reinforcing plate 42 can be avoided, and the problem of excessive turbulence intensity caused by the inability to block the transverse turbulence of the waste liquid flow due to the excessive narrowness of the reinforcing plate 42, which leads to waste liquid backflow, can be avoided. Second, when performing target exosome elution and recovery, the cleaning liquid spiral flow on the filter membrane surface causes tangential shear force due to the change in flow rate on the filter membrane surface and the impact of the spiral flow. If the shear force is too large, it will cause denaturation or fragmentation of the exosome membrane protein, thereby affecting the quality of the recovered exosome. Through this arrangement, when the cleaning liquid spiral flow impacts the surface of the first filter membrane 5, the reinforcing plate 42 can provide stable support, and when the cleaning liquid spiral flow impacts the first filter membrane 5, the relative motion between the first filter membrane 5 and the guide rib 41 can be inhibited, thereby reducing the additional shear force generated by vibration, effectively ensuring the qualified rate of the recovered exosome. If L2 is less than 2.3 times L1, the support of the reinforcing plate 42 is insufficient, and it cannot inhibit the vibration of the filter membrane in cooperation with the guide rib 41. The additional shear force generated by the vibration of the filter membrane and the shear force of the cleaning liquid spiral flow can easily exceed the critical value, causing a large number of exosomes to be broken during the recovery process. If L2 is greater than 3.5 times L1, the support performance of the reinforcing plate 42 is excessive, and the excessively wide reinforcing plate 42 will compress the number of waste liquid flow channels on the first guide member 4, causing the flow resistance of the waste liquid to increase and the flow rate to slow down during the purification and filtration process, which can easily lead to waste liquid backflow, causing impurities to deposit on the back of the first filter membrane 5, thereby reducing the flux of the first filter membrane 5 and affecting the purification efficiency. Only when the width L2 of the reinforcing plate 42 is 2.3-3.5 times the spacing between adjacent guide ribs 41 or the spacing between adjacent reinforcing plates 42 L1, can the quality of the recovered exosome be improved to 85% under the premise of ensuring the flow rate of the waste liquid and the flux of the filter membrane during the purification and filtration process.

[0051] Specifically, as Figure 8As shown, the second flow guide 7 is a center-holed plate structure, and the side in abutment with the second filter membrane 8 is a flow guide side, the outer peripheral edge of which is provided with a binding, and a plurality of straight linear protrusions 71 are uniformly arranged around the center. The protrusions include long edges 711 and short edges 712 arranged at intervals, one end of the long edges 711 extends to the binding, and the other end extends to the edge of the center through-hole, one end of the short edges 712 extends to the binding, and the other end extends to a predetermined distance from the center through-hole, and the included angle a between adjacent straight linear protrusions 71 is 5-15 degrees. It can be understood that the waste liquid flow channel is formed between adjacent straight linear protrusions 71, and the outer peripheral edge of the flow guide side is provided with a binding to guide the circumferential edge of the flow guide side to be provided with a planar area for bonding and fixing with the circumferential edge of the second filter membrane 8. Through this arrangement, first, better support for the second filter membrane 8 can be achieved, and by arranging the flow guide side in a radial structure with gradually tapered flow channel width, the long edges 711 and short edges 712 are more concentrated at one end closer to the center through-hole than at the other end, which can coordinate the support force for the second filter membrane 8 according to the negative pressure pressure zoning, avoiding the problem of deformation and collapse of the central region of the second filter membrane 8 caused by excessive negative pressure at the outlet position of the sealed channel 9; secondly, the long edges 711 and short edges 712 play a role in guiding waste liquid, and by arranging a gradually tapered flow channel, the waste liquid at the outer periphery of the flow guide side can be stably collected into the sealed channel 9, effectively reducing the probability of waste liquid accumulation at the outer periphery of the flow guide side, and during the waste liquid collection and flow process, as the flow channel width gradually tapers, the waste liquid is continuously accelerated, which is more conducive to the collection of waste liquid from the periphery to the center, further avoiding the problem of waste liquid accumulation at the flow guide side, and ensuring the filtration flux of the second filter membrane 8; thirdly, through the above arrangement, the flow channel width of the waste liquid flow channel suddenly changes when it approaches the inlet of the sealed channel 9, and when the waste liquid flow accelerated by the flow channel enters this area, the sudden increase in flow channel width causes the flow rate of the waste liquid flow to suddenly decrease and form a turbulent flow trend, and near the connection position of the sealed channel 9 and the second flow guide 7, the deposition of impurity particles is prone to occur due to the 90-degree change in flow direction, and the turbulent kinetic energy of the waste liquid flow can entrain the deposited cell fragments and protein aggregates in this area into the sealed channel 9 and discharge them, which can effectively reduce the deposition of impurity particles, further reducing the blockage rate of the purification device 24 in continuous operation for 24 hours, and the sudden decrease in flow rate of the waste liquid flow can reduce the boundary layer separation phenomenon in the sealed channel 9 caused by high-speed waste liquid jet, further ensuring the waste liquid transport efficiency and the filtration flux of the second filter membrane 8; finally, by limiting the included angle between adjacent straight linear protrusions 71, the relationship between waste liquid flow and support for the second filter membrane 8 is effectively balanced, which can avoid both the collapse problem caused by excessive deformation of the second filter membrane 8 and the flow channel blockage problem caused by excessively narrow flow channel width, or the accumulation problem caused by slow flow of waste liquid at the outer periphery of the flow guide side due to excessively wide flow channel width.

[0052] Specifically, as Figure 8As shown, the relationship between the distance K between the end of the short edge 712 and the center through hole and the included angle a between adjacent straight convex edges 71 satisfies: Wherein the unit of K is centimeter. The greater the included angle a, the smaller the distance K between the end of the short edge 712 and the center through hole. Firstly, by this setting, the waste liquid flow channel can be further optimized, and the deformation problem of the second filter membrane 8 in the use process can be avoided on the premise of ensuring the waste liquid conveying efficiency. Secondly, the negative pressure introduced into the second flow guide piece 7 is transmitted through the sealed channel 9 of the center through hole of the second flow guide piece 7, and finally acts on the surface of the second filter membrane 8. Because the second flow guide piece 7 has a certain distance from the center, the negative pressure will be gradually lost during transmission, resulting in that the negative pressure pressure received by the edge of the second flow guide piece 7 will be lower than that received by the center area of the second flow guide piece 7, and then the negative pressure suction received by the edge area of the second filter membrane 8 will be smaller, resulting in that the filtration flux of the edge area of the second filter membrane 8 will decrease. Through the above setting, a three-level negative pressure shunt transmission path of center through hole-short edge 712-edge is formed. The negative pressure is transmitted to the short edge 712 by the first shunt of the center through the long edge 711, and then the second shunt of the short edge 712 to the edge area of the second flow guide piece 7 is performed. By shunting to increase the independent negative pressure transmission path, the negative pressure is more uniform, which effectively reduces the local loss in the negative pressure transmission process. Further, the included angle between adjacent straight convex edges 71 determines the number of straight convex edges 71. The larger the included angle, the fewer the convex edges. On this basis, by setting a smaller distance between the end of the short edge 712 and the center through hole, the shunt can be guided in advance, and the vortex loss caused by the too wide flow channel during the transmission of the negative pressure from the long edge 711 to the short edge 712 can be effectively reduced. Through the above setting, the negative pressure can be transmitted to the edge area of the second flow guide piece 7 with low loss, and the filtration flux difference between the edge area and the center area of the second filter membrane 8 is less than 5%.

[0053] Specifically, as Figure 9As shown, the second flow guide 7 is a drainage side away from the second filter membrane 8, and the drainage side is provided with a plurality of arc-shaped ribs 72 evenly distributed around the center. One end of the arc-shaped rib 72 extends to the outer peripheral edge of the drainage side, and the other end extends to a predetermined distance from the center hole. The area S1 enclosed by the connecting line between the curvature centers of the arc-shaped ribs 72 is 0.3-0.7 times the area of the drainage side. Because there is a certain distance between the first filter membrane 5 and the second flow guide 7, when the high-speed sample to be purified flows between the basic filter part and the new filter part, the presence of the new filter part changes the flow path of the sample to be purified. When the sample fluid flows through the surface of the first filter membrane 5, a low-speed fluid layer close to the surface of the first filter membrane 5 is formed due to viscous force. The sample to be purified in the low-speed fluid layer cannot obtain enough kinetic energy, resulting in low lateral migration of impurity particles, which cannot quickly pass through the filter membrane surface, thereby reducing the filter membrane filtration flux and affecting the filtration efficiency. First, through this setting, the sample to be purified can form a spiral flow between the second flow guide 7 and the first filter membrane 5 to increase the kinetic energy of the sample to be purified, so that it can destroy the low-speed fluid layer on the surface of the first filter membrane 5, ensuring that the impurity particles have enough lateral migration energy, effectively improving the filtration flux of the first filter membrane 5. By extending one end of the arc-shaped rib 72 to a predetermined distance from the center hole, the sample to be purified can directly enter from the edge of the drainage side, which can further ensure orderly flow and avoid energy loss. Second, by limiting the area enclosed by the connecting line between the curvature centers of the arc-shaped ribs 72, the bending degree of the arc-shaped ribs 72 can guide the sample to be purified to form a stable spiral flow, which can sufficiently destroy the low-speed fluid layer and effectively reduce the energy loss of the sample to be purified. If the area enclosed by the connecting line between the curvature centers of the arc-shaped ribs 72 is less than 0.3 times the area of the drainage side, the arc-shaped ribs 72 are too curved, so that the spiral flow can only cover a small area of the surface of the first filter membrane 5, resulting in a decrease in the local filtration flux of the first filter membrane 5. If the area enclosed by the connecting line between the curvature centers of the arc-shaped ribs 72 is greater than 0.7 times the area of the drainage side, the arc-shaped ribs 72 are too small, which cannot guide the sample to be purified to form a spiral flow, resulting in the inability to destroy the low-speed fluid layer and the problem of reduced filter membrane filtration flux.

[0054] Specifically, in order to further improve the purification efficiency, the shell 12 comprises a left shell and a right shell, and the left shell and the right shell are both provided with a basic filter part and a new filter part, the new filter part in the left shell is oppositely arranged with the new filter part in the right shell and is spaced apart by a certain distance, and the liquid inlet and outlet interface 1 is arranged at the lower part of the shell 12 and is communicated to the area between the two new filter parts. The left shell and the right shell are both provided with a negative pressure interface 3, and the two negative pressure interfaces 3 are both arranged at the lower part of the shell 12. It should be pointed out that the purification device described in the present application is used vertically during purification, so the negative pressure interface 3 is at the bottom during use, which can effectively reduce the retention rate of waste liquid in the shell 12. The left shell and the right shell are integrally sealed by high-strength structural adhesive or by ultrasonic welding. After the purified sample is injected into the gap between the left shell and the right shell through the liquid inlet and outlet interface 1, the left shell negative pressure interface 3 and the right shell negative pressure interface 3 are respectively connected to the negative pressure source. Under the driving of negative pressure, the basic filter unit and the new filter unit in the two side shells simultaneously perform exosome purification.

[0055] Finally, it should be pointed out that: the above embodiments are only used to illustrate the technical solutions of the present application, and not to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. An exosome purification device, characterized by, The application relates to a filter device. The filter device comprises a shell, an inlet-outlet interface, a flushing interface and a negative pressure interface; A basic filtering part is arranged in the inner cavity of the shell, and comprises a first flow guide member and a first filter membrane covering one side of the first flow guide member away from the inner side wall of the shell, the first flow guide member is configured to allow filtered liquid to pass through, the first filter membrane and the first flow guide member are both provided with through holes in the center, and the region between the first flow guide member and the inner side wall of the shell is configured as a negative pressure drainage cavity, the negative pressure interface is connected to the negative pressure drainage cavity, and the peripheral edge of the first filter membrane is fixedly connected to the peripheral edge of the first flow guide member. An additional filtering part is arranged on the side of the first filter membrane away from the first flow guide member, and comprises a second flow guide member and a second filter membrane covering one side of the second flow guide member away from the first filter membrane, the second flow guide member is arranged at a predetermined distance from the first flow guide member, and is configured to prevent filtered liquid from passing through, the second flow guide member is provided with a through hole in the center, a sealed channel for conveying filtered liquid of the additional filtering part is arranged between the first flow guide member and the second flow guide member, one end of the sealed channel is connected to the through hole in the center of the first flow guide member, the other end of the sealed channel is connected to the through hole in the center of the second flow guide member, the first filter membrane is sleeved on the outer periphery of the sealed channel, the peripheral edge of the second filter membrane is fixedly connected to the peripheral edge of the second flow guide member, and the peripheral edge of the additional filtering part is spaced apart from the inner side wall of the shell by a certain distance.

2. The exosome purification device of claim 1, wherein, The peripheral side of the negative pressure drainage cavity is provided with a supporting boss for supporting the basic filtering part, and the peripheral edge of the first flow guide member abuts against the supporting boss. The negative pressure drainage cavity is further provided with a drainage plate, the drainage plate is provided with a honeycomb-shaped through hole penetrating through the upper and lower surfaces of the drainage plate, and the drainage plate is spaced apart from the first flow guide member by a certain distance.

3. The exosome purification device of claim 1, wherein, The first flow guide member and the second flow guide member have the same diameter, the cross-sectional shape of the sealed channel is circular, and the diameter D1 of the sealed channel is 0.02-0.4 times the diameter of the first flow guide member or the second flow guide member.

4. The exosome purification device of claim 3, wherein, The vertical distance L3 between the second flow guide member and the first flow guide member is 0.02-1.5 times the diameter D1 of the sealed channel.

5. The exosome purification device of any one of claims 1-4, wherein, The peripheral edge of the first flow guide member is treated by edge covering, the first flow guide member is provided with a plurality of arrayed flow guide ribs on the side abutting against the first filter membrane, and the other side close to the negative pressure drainage cavity is provided with a plurality of arrayed reinforcing plates abutting against the flow guide ribs, the distance between adjacent flow guide ribs is the same as the distance between adjacent reinforcing plates, and the length extension directions of the flow guide ribs and the reinforcing plates are perpendicular to each other, and the flow guide ribs and the reinforcing plates are staggered to form a channel for filtered liquid to pass through.

6. The exosome purification device of claim 5, wherein, The width L2 of the reinforcing plate is 2.3-3.5 times the distance between adjacent flow guide ribs or the distance L1 between adjacent reinforcing plates.

7. The exosome purification device of claim 6, wherein, The second flow guide is a plate structure with a central hole, and the side abutting against the second filter membrane is a flow guide side, the outer peripheral edge of the flow guide side is provided with a binding edge, and a plurality of straight linear protrusions are uniformly arranged around the center, the protrusions include long protrusions and short protrusions arranged at intervals, one end of the long protrusion extends to the binding edge, and the other end extends to the edge of the central hole, one end of the short protrusion extends to the binding edge, and the other end extends to a predetermined distance from the central hole, and the included angle α between adjacent straight linear protrusions is 5-15 degrees.

8. The exosome purification device of claim 7, wherein, The relationship between the distance K between the short edge end and the center through hole and the included angle α between the adjacent straight convex edges satisfies: wherein the unit of K is centimeter.

9. The exosome purification device of claim 8, wherein, The side away from the second filter membrane of the second flow guide is a flow guide side, the flow guide side is provided with a plurality of arc-shaped protrusions uniformly arranged around the center, one end of the arc-shaped protrusion extends to the outer peripheral edge of the flow guide side, and the other end extends to a predetermined distance from the central hole, and the area S1 enclosed by the lines connecting the centers of curvature of the arc-shaped protrusions is 0.3-0.7 times the area of the flow guide side.

10. The exosome purification device of claim 1, wherein, The shell comprises a left shell and a right shell, the left shell and the right shell are both provided with a basic filter part and a new filter part, the new filter part in the left shell is oppositely arranged with the new filter part in the right shell and is spaced apart by a certain distance, and the liquid inlet and outlet interface is arranged at the lower part of the shell and is communicated to the region between the two new filter parts. The left shell and the right shell are both provided with a negative pressure interface, and the two negative pressure interfaces are both arranged at the lower part of the shell.

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