Search

CN-121991786-A - Exosome microvesicle harmless separation purification device

CN121991786ACN 121991786 ACN121991786 ACN 121991786ACN-121991786-A

Abstract

The invention belongs to the technical field of exosome processing, and discloses a nondestructive separation and purification device for microvesicles of an exosome, which comprises a sealed separation box, wherein a lamination assembly is arranged in an inner cavity of the sealed separation box, the lamination assembly is sleeved outside a central shaft assembly, and a pressing plate is matched with the top end of the lamination assembly; and (5) synchronously driving the components. According to the invention, when the micro-nano groove is in a compressed state, an accurate nano channel is formed for interception and filtration, and when the micro-nano groove is in a relaxed state, the elastic sheet rebounds rapidly along with the release of axial pressure, the periodical geometric deformation, namely the deformation of the geometric shape of the section of the micro-nano groove, breaks the mechanical stability of a filter cake layer from a physical layer, so that particles which are stuck in the groove or adsorbed on the surface lose clamping force instantaneously, and automatically fall off along with the shearing action of surface fluid, and the flux attenuation rate is controlled to be a smaller value under the long-time operation of the device, so that continuous and stable filtration in a real sense is realized.

Inventors

  • WEI XIN
  • DENG GUANGWEN
  • WU SHUXIA
  • TAN JUN

Assignees

  • 艾一生命科技(广东)有限公司

Dates

Publication Date
20260508
Application Date
20260108

Claims (10)

  1. 1. The nondestructive separation and purification device for the exosome microvesicles is characterized by comprising a sealing separation box (1), wherein a lamination assembly (5) is arranged in an inner cavity of the sealing separation box, the lamination assembly (5) is sleeved outside a central shaft assembly (6), and a pressing plate (9) is matched with the top end of the lamination assembly (5); The synchronous driving assembly (8) is arranged at the top end of the sealing separation box (1), is in transmission connection with the pressure plate (9) and the central shaft assembly (6) and is used for driving the pressure plate (9) and the central shaft assembly (6) to reciprocate along the axial direction; The suction assembly (7) is arranged at the bottom end of the sealed separation box (1) and is fixedly connected with the bottom end of the central shaft assembly (6); The lamination assembly (5) comprises a plurality of elastic sheets (501) stacked along the axial direction, and micro-nano grooves (502) are formed in the surfaces of the elastic sheets (501); when the synchronous driving assembly (8) drives the pressure plate (9) to press down, adjacent elastic sheets (501) are mutually pressed so as to enable the micro-nano grooves (502) to be closed to form a filtering channel; When the synchronous drive assembly (8) releases pressure, the elastic sheet (501) is rebound and reset to expand the channel section of the micro-nano groove (502); the suction assembly (7) is configured to apply a negative pressure to the interior of the central shaft assembly (6) while the lamination assembly (5) is rebound.
  2. 2. The device for non-destructive separation and purification of exosome microvesicles according to claim 1, wherein a circle of limiting convex rings (503) are circumferentially arranged at the outer edge of the elastic sheet (501), and the axial height of the limiting convex rings (503) is greater than the depth of the micro-nano grooves (502); When the lamination assembly (5) is in a pressed state, the bottom surface of the upper elastic sheet (501) is abutted against the top surface of the limiting convex ring (503) of the lower elastic sheet (501), and a filtering gap with constant height is defined between two adjacent elastic sheets (501).
  3. 3. The device for non-destructive separation and purification of exosome microvesicles according to claim 1, wherein the central shaft assembly (6) comprises a main shaft (601) with a hollow structure and an extension shaft (603) connected to the bottom end of the main shaft (601); A plurality of diversion holes (602) are formed in the pipe wall of the main shaft (601), and the diversion holes (602) are communicated with the inner side of the lamination assembly (5) and the inner cavity of the main shaft (601); the extension shaft (603) penetrates through the bottom of the sealed separation box (1) and is connected with the suction assembly (7).
  4. 4. The device for non-destructive separation and purification of exosome microvesicles according to claim 3, wherein the suction assembly (7) comprises a suction tube (701) and a piston plate (702) slidably and sealingly disposed inside the suction tube (701), the piston plate (702) is fixedly connected with the extension shaft (603) so as to follow synchronous displacement of the central shaft assembly (6), and an inner cavity of the suction tube (701) is communicated with the top end of the main shaft (601) through a pipeline.
  5. 5. The device for non-destructive separation and purification of exosome microvesicles according to claim 4, wherein the suction tube (701) is communicated with a liquid discharge tube (706), a one-way valve is installed inside the liquid discharge tube (706), a return spring (704) is disposed between the inner bottom wall of the suction tube (701) and the piston plate (702), and the return spring (704) is used for applying a thrust force to the piston plate (702) towards a return direction.
  6. 6. The device for non-destructive separation and purification of exosome microvesicles according to claim 1, wherein the micro-nano grooves (502) are concentrically or spirally distributed on the upper surface of the elastic sheet (501) along the radial direction of the elastic sheet (501), the cross section of the micro-nano grooves (502) is in a V-shaped structure or a trapezoid structure, and the depth of the micro-nano grooves (502) is gradually reduced from outside to inside along the radial direction.
  7. 7. The device for non-destructive separation and purification of exosome microvesicles according to claim 1, wherein the synchronous driving assembly (8) comprises a frame (804), a power shaft (803) rotatably mounted on the frame (804) and a cam (805) fixedly sleeved on the power shaft (803); The pressure plate (9) or the filtrate valve (10) is connected with a synchronous frame (806), and the outer contour surface of the cam (805) is always kept in abutting contact with the stress surface of the synchronous frame (806); when the cam (805) rotates, the synchronous frame (806) is driven to drive the pressure plate (9) to reciprocate along the vertical direction.
  8. 8. The device for nondestructively separating and purifying the exosome microvesicles is characterized in that a uniform dispersion pipe (12) is communicated with the center of the bottom end of the sealed separation box (1), the uniform dispersion pipe (12) is of a funnel-shaped flaring structure, a raw liquid inlet valve (11) is connected to the inlet of the bottom end of the uniform dispersion pipe, and concentrated liquid valves (2) are symmetrically communicated with the two sides of the upper part of the outer wall of the sealed separation box (1).
  9. 9. The device for non-destructive separation and purification of exosome microvesicles according to claim 8, wherein the output ends of the concentrate valves (2) at both sides are respectively connected with a collecting pipe (3), and the tail ends of the two collecting pipes (3) are commonly connected to a collecting valve (4) for guiding out separated enriched liquid.
  10. 10. The device for non-destructive separation and purification of exosome microvesicles according to claim 1, wherein the surface of the elastic sheet (501) is grafted with a polyethylene glycol hydrophilic coating or a zwitterionic polymer coating, and the elastic sheet (501) is made of medical grade high polymer material with a Shore A hardness of 50-70 degrees.

Description

Exosome microvesicle harmless separation purification device Technical Field The invention belongs to the technical field of exosome processing, and particularly relates to a nondestructive separation and purification device for exosome microvesicles. Background The exosome is a disc-shaped vesicle secreted by cells and having a diameter of 30-150 nm, and the inside of the vesicle is wrapped with bioactive substances such as proteins, nucleic acids and the like, so that the exosome has extremely high application value in intercellular communication, disease diagnosis and drug carrier development. At present, the separation and purification of exosomes are major bottlenecks limiting their large-scale clinical application. Traditional extraction methods mainly include ultracentrifugation, polymer precipitation, and conventional membrane filtration (e.g., dead-end filtration or cross-flow filtration). Among them, the ultracentrifugation method is expensive in equipment, takes very long time (often more than 4 hours), and extremely high centrifugal force (often more than 100,000 g) easily causes irreversible physical damage or aggregation of exosome membrane structures, and the polymer precipitation method easily introduces chemical impurities which are difficult to remove, affecting the accuracy of subsequent biological experiments. In the prior art, the filtration and separation technology based on the microporous membrane is widely focused due to the simple operation and easy amplification, but has serious problems of membrane pollution and pore diameter blockage in practical application. In the conventional microporous filter membrane (such as PES and PVDF membrane), the pore size structure is static, when filtering biological samples rich in proteins and colloids, particles larger than the pore size and nonspecifically adsorbed proteins can rapidly accumulate on the surface of the membrane to form a compact 'filter cake layer', so that the transmembrane pressure difference (TMP) rises sharply, and in order to maintain the filtering efficiency, operators often have to apply higher driving pressure, which not only increases energy consumption, but also is more serious, the originally softer exosomes are forced to squeeze and deform or even break by high pressure, so that the exosomes are forced to enter the filtrate side through the filter pores, and the purity is reduced and the samples are lost. In addition, the existing exosome continuous purification device often lacks effective hydrodynamic coupling design, and the export of dialysate mainly relies on natural osmosis or independent pump source, leads to easy back pressure retention phenomenon of the interior of the central liquid collecting tube, has reduced effective filtration pressure difference. How to design a nondestructive separation device which can not only fundamentally solve the problem of filter hole blockage from physical mechanism, but also furthest protect the integrity of an exosome lipid bilayer and has a self-driven fluid conveying function is a key technical problem to be solved urgently in the technical field of exosome processing at present. Disclosure of Invention The invention aims to provide a nondestructive separation and purification device for exosome microvesicles, which aims to solve the problems in the background technology. In order to achieve the aim, the invention provides the technical scheme that the device for nondestructively separating and purifying the exosome microvesicles comprises a sealed separation box, wherein a lamination assembly is arranged in an inner cavity of the sealed separation box, the lamination assembly is sleeved outside a central shaft assembly, and a pressing plate is matched with the top end of the lamination assembly; The synchronous driving assembly is arranged at the top end of the sealing separation box, is in transmission connection with the pressure plate and the central shaft assembly and is used for driving the pressure plate and the central shaft assembly to reciprocate along the axial direction; The suction assembly is arranged at the bottom end of the sealed separation box and is fixedly connected with the bottom end of the central shaft assembly; the lamination assembly comprises a plurality of elastic sheets which are axially stacked, and micro-nano grooves are formed in the surfaces of the elastic sheets; When the synchronous driving assembly drives the pressure plate to press down, adjacent elastic sheets are mutually pressed so that the micro-nano grooves are closed to form a filtering channel; when the synchronous driving assembly releases pressure, the elastic sheet is rebound and reset to expand the channel section of the micro-nano groove; the suction assembly is configured to apply a negative pressure to the interior of the center shaft assembly while the lamination assembly is rebound and reset. As a further technical scheme of the invention, a circle of limiting convex r