CN212102818U - Concentrated exosome filter equipment - Google Patents

Abstract

The utility model discloses a concentrated exosome filter device, which comprises an outer sleeve test tube, wherein a coarse filter tube and an ultrafiltration tube are inserted in the outer sleeve test tube, the ultrafiltration tube is detachably connected to the lower end of the coarse filter tube through a middle connecting sleeve, a liquid collecting cavity is formed between the ultrafiltration tube and the outer sleeve test tube, a mesh supporting layer is arranged in the middle of the middle connecting sleeve between the coarse filter tube and the ultrafiltration tube, and a coarse filter membrane is arranged at the upper end of the mesh supporting layer; the lower extreme of ultrafiltration pipe is provided with lower part adapter sleeve, and lower part adapter sleeve can dismantle and be connected with the ultrafiltration lid, and a plurality of small openings have been seted up to the bottom of ultrafiltration lid, is provided with the milipore filter in the ultrafiltration lid, and the ultrafiltration lid is provided with antiseized even mesh disk in the upper end of milipore filter, and antiseized even mesh disk diameter is less than the internal diameter of ultrafiltration lid and can freely horizontal migration inside the ultrafiltration lid. The utility model discloses a two-stage filtration effectively reduces large granule protein pollution to can prevent effectively that the exosome from adhering to the milipore filter, prevent that the milipore filter from blockking up and exosome loss.

Description

Concentrated exosome filter equipment

Technical Field

The utility model relates to a molecular biology apparatus technical field, in particular to concentrated exosome filter equipment.

Background

Exosomes (exosomes) are nanoscale membrane vesicles which are released into an extracellular environment after fusion of multivesicular endosomes (MVBs) of eukaryotic cells and cell membranes, have a lipid bilayer membrane structure, have the size of about 40-150 nm, and are widely distributed in body fluid; can be secreted by various cells, and contains various biologically active substances such as protein, lipid, mRNAs, microRNAs, signal molecules and the like; has stronger targeting property, is easy to fuse with the cell membrane of a target cell, selectively delivers biological active substances to the target cell, carries out information transmission among different cells, regulates signal conduction among the cells and plays a plurality of biological functions.

The extraction method of the exosome at present can be mainly summarized as the following methods: ultracentrifugation, polymer precipitation, magnetic bead, and ultrafiltration. The ultrafiltration method is to filter the solvent and small molecular substances to the other side of the membrane and to retain the relatively large molecular substances on the ultrafiltration membrane, so as to achieve the purpose of separation. The diameter range of the exosome is 40-150 nm, the exosome is larger than that of a common protein, and the exosome in the sample can be separated by using an ultrafiltration membrane with the pore diameter smaller than 150nm for low-speed long-time centrifugation. The advantages are that: the technology has the advantages of simple and rapid operation, low cost, convenient use and high enrichment efficiency. However, large-particle protein is seriously polluted, the possibility of blocking an ultrafiltration membrane by attaching exosomes is high, and the exosomes are easily attached to the membrane and lost.

SUMMERY OF THE UTILITY MODEL

The utility model aims at providing a concentrated exosome filter equipment adopts the two-stage filtration, effectively reduces large granule protein pollution to can effectively prevent exosome attached ultrafiltration membrane, prevent that the ultrafiltration membrane from blockking up and exosome loss.

The above technical purpose of the present invention can be achieved by the following technical solutions: a concentrated exosome filter device comprises a hollow outer sleeve test tube with an opening at the upper end, a coarse filter tube and an ultrafiltration tube which are both open at two ends are inserted into the outer sleeve test tube, the ultrafiltration tube is detachably connected to the lower end of the coarse filter tube through a middle connecting sleeve, a liquid collection cavity is formed between the lower end of the ultrafiltration tube and the bottom of the outer sleeve test tube, the edge of the upper end of the coarse filter tube extends outwards to form a support ring erected on the opening end of the outer sleeve test tube, a mesh support layer is arranged between the coarse filter tube and the ultrafiltration tube in the middle of the middle connecting sleeve, and a detachable coarse filter membrane is arranged at the upper end of the mesh support layer;

the lower extreme of ultrafiltration pipe is provided with sub-unit connection sleeve, sub-unit connection sleeve can dismantle and be connected with the ultrafiltration lid, a plurality of small openings have been seted up to the bottom of ultrafiltration lid, be provided with the milipore filter that can follow the trade in the ultrafiltration lid, in the ultrafiltration lid the upper end of milipore filter is provided with antiseized even mesh disk, antiseized even mesh disk diameter is less than the internal diameter of ultrafiltration lid just can be at the inside free horizontal migration of ultrafiltration lid.

By adopting the technical scheme, the processed sample is added into the coarse filtration tube, and then the jacketed test tube is placed into a centrifuge for low-speed centrifugal filtration. In the centrifugal filtration process, most of microvesicles in a sample are retained in a coarse filtration tube by a coarse filtration membrane, and exosomes, soluble proteins, high-abundance proteolytic fragments, smaller particles and the like enter an ultrafiltration tube through the coarse filtration membrane;

in the centrifugal filtration process, because the device is in the pivoting state for the inner wall that anti-adhesion mesh disk hugs closely the ultrafiltration lid always rolls, and the in-process that rolls, the mesh of anti-adhesion mesh disk constantly scrapes the exosome of adhesion on the ultrafiltration membrane, prevents effectively that exosome adhesion from blockking up the ultrafiltration membrane on the ultrafiltration membrane, improves filter speed, and effectively reduces the loss of exosome.

The utility model discloses a further set up to: the lower end of the lower connecting sleeve is provided with a limiting ring extending to the anti-adhesion mesh wafer.

By adopting the technical scheme, the anti-adhesion mesh wafer is prevented from tilting in the rolling process.

The utility model discloses a further set up to: the inner two ends of the middle connecting sleeve are provided with internal threads, the lower end of the coarse filtering pipe is provided with external threads connected with the middle connecting sleeve in a threaded manner, and the upper end of the ultrafiltration pipe is provided with external threads connected with the middle connecting sleeve in a threaded manner.

The utility model discloses a further set up to: the inner wall of the ultrafiltration cover is provided with internal threads, and the outer wall of the lower connecting sleeve is provided with external threads in threaded connection with the ultrafiltration cover.

The utility model discloses a further set up to: the overcoat test tube in the lower extreme of ultrafiltration lid is provided with the round and keeps off the ring.

Through adopting above-mentioned technical scheme, prevent that coarse filtration pipe and ultrafiltration pipe from directly falling into overcoat test tube bottom to be convenient for take out.

The utility model discloses a further set up to: the lateral wall of thick filter tube upper end has seted up first air vent, the second air vent has been seted up to the lateral wall of ultrafiltration tube upper end, the lateral wall of overcoat test tube in first air vent, the second air vent reaches the position department that the collection liquid chamber upper end corresponds all has seted up the third air vent.

Through adopting above-mentioned technical scheme, maintain inside and outside atmospheric pressure balanced, the unbalanced influence of atmospheric pressure filtration speed when preventing to filter.

The utility model discloses a further set up to: the aperture of the coarse filtering membrane is 130-150 nm.

The utility model discloses a further set up to: the aperture of the ultrafiltration membrane is 40-50 nm.

To sum up, the utility model discloses following beneficial effect has:

firstly, two-stage filtration is adopted, wherein the first stage filters large protein molecules and large particle impurities, and the second stage filters small protein molecules and small particle impurities, so that exosomes with higher purity are finally left, and the large particle protein pollution is effectively reduced;

and secondly, the anti-adhesion mesh wafer continuously rolls in the centrifugal process to scrape the exosomes on the ultrafiltration membrane, so that the exosomes are prevented from being adhered on the ultrafiltration membrane to block the ultrafiltration membrane, the filtering and extracting speed is improved, no exosomes are adhered on the ultrafiltration membrane, and the loss of the exosomes is reduced.

Drawings

Fig. 1 is an exploded view of the present invention;

fig. 2 is a cross-sectional view of the present invention.

In the figure: 1. sleeving a test tube; 11. a liquid collection cavity; 12. a baffle ring; 2. a coarse filter pipe; 21. a support ring; 3. an ultrafiltration tube; 4. the middle part is connected with a sleeve; 41. a mesh support layer; 42. a coarse filtration membrane; 5. the lower part is connected with a sleeve; 51. a limiting ring; 6. an ultrafiltration cover; 61. a leak hole; 7. ultrafiltration membranes; 8. anti-adhesion mesh wafers; 91. a first vent hole; 92. a second vent hole; 93. and a third vent hole.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings.

The embodiment is a concentrated exosome filtering device, and referring to fig. 1-2, the device comprises a hollow outer sleeve test tube 1 with an opening at the upper end, a coarse filter tube 2 and an ultrafiltration tube 3 which are both open at two ends are inserted into the outer sleeve test tube 1, the ultrafiltration tube 3 is detachably connected to the lower end of the coarse filter tube 2 through a middle connecting sleeve 4, internal threads are arranged at two ends inside the middle connecting sleeve 4, external threads in threaded connection with the middle connecting sleeve 4 are arranged at the lower end of the coarse filter tube 2, and external threads in threaded connection with the middle connecting sleeve 4 are arranged at the upper end of the ultrafiltration tube 3; a liquid collecting cavity 11 is formed between the lower end of the ultrafiltration tube 3 and the bottom of the outer sleeve test tube 1, a mesh supporting layer 41 is arranged in the middle of the middle connecting sleeve 4 between the coarse filtration tube 2 and the ultrafiltration tube 3, a detachable coarse filtration membrane 42 is arranged at the upper end of the mesh supporting layer 41, and the aperture of the coarse filtration membrane 42 is 140nm and used for filtering coarse impurities and large proteins in a stock solution.

The lower extreme of ultrafiltration pipe 3 is provided with a lower part adapter sleeve 5, lower part adapter sleeve 5 can dismantle and be connected with an ultrafiltration lid 6, the inner wall of ultrafiltration lid 6 is provided with the internal thread, the outer wall of lower part adapter sleeve 5 is provided with the external screw thread with ultrafiltration lid 6 threaded connection, a plurality of small openings 61 have been seted up to ultrafiltration lid 6's bottom, be provided with one in the ultrafiltration lid 6 and follow the milipore filter 7 who trades, the aperture of milipore filter 7 is 45nm, be used for filtering the protein molecule that the diameter ratio exosome is little, be provided with an antiseized even mesh disk 8 in ultrafiltration lid 6's upper end, the lower extreme of sub-adapter sleeve 5 is provided with the spacing collar 51 that the round extended to antiseized even mesh disk 8, mesh disk 8 diameter is less than the internal diameter of antiseized even filter lid 6 and can.

The edge of 2 upper ends of thick filter tube outwards extends and forms the round and sets up support ring 21 on 1 open end of overcoat test tube, and overcoat test tube 1 is provided with the round in the lower extreme of ultrafiltration lid 6 and keeps off ring 12, prevents that thick filter tube 2 and ultrafiltration tube 3 from directly falling into 1 bottom of overcoat test tube to be convenient for take out.

A first air vent 91 has been seted up to the lateral wall of 2 upper ends of thick filter tube, and a second air vent 92 has been seted up to the lateral wall of ultrafiltration tube 3 upper end, and a third air vent 93 has all been seted up in the position department that first air vent 91, second air vent 92 and collection liquid chamber 11 upper end correspond to the lateral wall of overcoat test tube 1, maintains inside and outside atmospheric pressure balanced, and the unbalanced air pressure influences the filter speed during preventing to filter.

The use method comprises the following steps: the processed sample is added to the crude filter tube 2, and the jacketed test tube 1 is then placed in a centrifuge for low-speed centrifugation. In the centrifugal filtration process, most of the microvesicles in the sample are retained in the coarse filtration tube 2 by the coarse filtration membrane 42, and the exosomes, soluble proteins, high-abundance proteolytic fragments and finer particles and the like enter the ultrafiltration tube 3 through the coarse filtration membrane 42, and in the centrifugal process, the soluble proteins, the high-abundance proteolytic fragments and the finer particles in the sample are filtered out from the ultrafiltration membrane 7 on the side wall and enter the liquid collection cavity 11, so that the exosomes with higher purity are obtained in the ultrafiltration tube 3;

in the centrifugal filtration process, because the device is in the pivoting state for the inner wall that ultrafiltration lid 6 was hugged closely always to antiseized mesh disk 8 rolls, and the in-process that rolls, the mesh of antiseized mesh disk 8 is constantly scraped the exosome of adhesion on milipore filter 7, prevents effectively that exosome adhesion from blockking up milipore filter 7 on milipore filter 7, improves filter speed, and effectively reduces the loss of exosome.

The present embodiment is only for explaining the present invention, and it is not limited to the present invention, and those skilled in the art can make modifications to the present embodiment without inventive contribution as required after reading the present specification, but all of them are protected by patent laws within the scope of the claims of the present invention.

Claims (8)

1. The utility model provides a concentrated exosome filter equipment, includes cavity upper end open-ended overcoat test tube (1), its characterized in that: a coarse filter tube (2) and an ultrafiltration tube (3) both with two open ends are inserted in the outer sleeve tube (1), the ultrafiltration tube (3) is detachably connected to the lower end of the coarse filter tube (2) through a middle connecting sleeve (4), a liquid collecting cavity (11) is formed between the lower end of the ultrafiltration tube (3) and the bottom of the outer sleeve tube (1), the edge of the upper end of the coarse filter tube (2) extends outwards to form a support ring (21) which is arranged on the open end of the outer sleeve tube (1), a mesh support layer (41) is arranged in the middle of the middle connecting sleeve (4) between the coarse filter tube (2) and the ultrafiltration tube (3), and a detachable coarse filter membrane (42) is arranged at the upper end of the mesh support layer (41);

the lower extreme of ultrafiltration tube (3) is provided with sub-unit connection sleeve pipe (5), sub-unit connection sleeve pipe (5) can be dismantled and be connected with ultrafiltration lid (6), a plurality of small openings (61) have been seted up to the bottom of ultrafiltration lid (6), be provided with in ultrafiltration lid (6) and follow ultrafiltration membrane (7) of trading, in ultrafiltration lid (6) the upper end of ultrafiltration membrane (7) is provided with antiseized even mesh disk (8), antiseized even mesh disk (8) diameter is less than the internal diameter of ultrafiltration lid (6) just can be in the inside free horizontal migration of ultrafiltration lid (6).

2. The concentrated exosome filtration device according to claim 1, characterized in that: the lower end of the lower connecting sleeve (5) is provided with a limiting ring (51) extending to the anti-adhesion mesh wafer (8).

3. The concentrated exosome filtration device according to claim 1, characterized in that: the inside both ends of middle part adapter sleeve (4) all are provided with the internal thread, the lower extreme of thick filter tube (2) be provided with middle part adapter sleeve (4) threaded connection's external screw thread, the upper end of ultrafiltration pipe (3) be provided with middle part adapter sleeve (4) threaded connection's external screw thread.

4. The concentrated exosome filtration device according to claim 1, characterized in that: the inner wall of the ultrafiltration cover (6) is provided with internal threads, and the outer wall of the lower connecting sleeve (5) is provided with external threads connected with the ultrafiltration cover (6) in a threaded manner.

5. The concentrated exosome filtration device according to claim 1, characterized in that: the outer sleeve test tube (1) is provided with a ring of baffle ring (12) at the lower end of the ultrafiltration cover (6).

6. The concentrated exosome filtration device according to claim 1, characterized in that: first air vent (91) have been seted up to the lateral wall of thick filter tube (2) upper end, second air vent (92) have been seted up to the lateral wall of ultrafiltration tube (3) upper end, the lateral wall of overcoat test tube (1) in first air vent (91), second air vent (92) and position department that collection liquid chamber (11) upper end corresponds has all seted up third air vent (93).

7. The concentrated exosome filtration device according to claim 1, characterized in that: the pore diameter of the coarse filtering membrane (42) is 130-150 nm.

8. The concentrated exosome filtration device according to claim 1, characterized in that: the aperture of the ultrafiltration membrane (7) is 40-50 nm.

Images