CN214486404U

CN214486404U - Exosome extraction centrifuge tube combining ultrafiltration method and ultracentrifugation method

Info

Publication number: CN214486404U
Application number: CN202022535894.8U
Authority: CN
Inventors: 周卿
Original assignee: Huaxiayuan Cell Engineering Group Co Ltd
Current assignee: Huaxiayuan Cell Engineering Group Co Ltd
Priority date: 2020-11-05
Filing date: 2020-11-05
Publication date: 2021-10-26
Anticipated expiration: 2030-11-05

Abstract

The utility model relates to a molecular biology and clinical examination technical field specifically are exosome of combination ultrafiltration method and ultracentrifugation method draws centrifuging tube, including the milipore filter in body, tube cap and the body. The ultrafiltration membrane is arranged in the tube body in an inverted cone shape, and the top of the ultrafiltration membrane is embedded into a groove at the bottom of the tube body; the inner wall of the pipe body is provided with a bulge for fixing the upper edge of the ultrafiltration membrane. By combining an ultracentrifugation method and an ultrafiltration method, the ultrafiltration membrane is set to be in an inverted cone shape to extract exosomes by centrifuging cell supernatant, so that the structure of the tube body and the steps of exosome extraction are simplified, and the exosome extraction efficiency is improved.

Description

Exosome extraction centrifuge tube combining ultrafiltration method and ultracentrifugation method

Technical Field

The utility model relates to a molecular biology and clinical examination technical field specifically are exosome extraction centrifuging tube that combines ultrafiltration method and ultracentrifugation method.

Background

Exosome is a vesicle secreted by living cells, has a diameter of 40-150nm and a double-layer lipid membrane structure, and is widely present in various body fluids such as peripheral blood, ascites, urine, saliva, cerebrospinal fluid and the like. At present, mainly used exosome extraction methods are an ultracentrifugation method, an ultrafiltration method, a density gradient centrifugation method, a kit extraction method, an immunomagnetic bead method and the like, but the ultracentrifugation method and the ultrafiltration method are more commonly applied.

The ultracentrifugation process generally comprises 4 steps: (1) centrifuging at low speed to remove residual cells; (2) removing cell debris by high-speed centrifugation; (3) filtering with 0.22 μm filter membrane to remove large diameter vesicles; (4) precipitating the exosome by ultracentrifugation, and selecting sucrose density gradient centrifugation for improving the purity. However, the ultracentrifugation method also has disadvantages of long operation time, low separation efficiency, expensive instrument, and the like.

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 40-150nm, the most common filtering membrane has the pore diameter of 0.8 μm, 0.45 μm or 0.22 μm, can be used for collecting substances larger than 800nm, 400nm or 200nm, and is also designed into a micro-column porous silicon cilia structure to separate 40-100nm exosomes. However, the ultrafiltration method is likely to cause accumulation of various substances such as dead cell residues and proteins on the filtration membrane, and thus has a high possibility of clogging, which affects filtration efficiency, and further causes adhesion of exosomes, thereby causing loss. Furthermore, the pressure and shear forces during filtration may damage the exosomes by deformation.

At present, the centrifugal filter device on the market has a complex structure, is not convenient enough to produce, is easy to damage the exosome to be extracted, and has low extraction efficiency.

SUMMERY OF THE UTILITY MODEL

To the problem, the utility model provides an exosome of combining ultrafiltration method and ultracentrifugation method draws centrifuging tube combines ultracentrifugation method and ultrafiltration method, realizes simplifying the exosome and draws the purpose of step and body structure.

The utility model provides a technical scheme as follows: an exosome extraction centrifuge tube combining an ultrafiltration method and an ultracentrifugation method comprises a tube body, a tube cover and an ultrafiltration membrane in the tube body; the pore diameter of the ultrafiltration membrane is between 0.15 and 0.30 mu m, the ultrafiltration membrane is arranged in the tube body in an inverted cone shape, and the top of the ultrafiltration membrane is embedded into a groove port at the bottom of the tube body; the inner wall of the pipe body is provided with a bulge for fixing the upper edge of the ultrafiltration membrane, so that the ultrafiltration membrane can be prevented from falling off when the centrifugal machine rotates.

Further as the improvement of the technical proposal of the utility model, the ultrafiltration membrane is an ultrafiltration membrane with the aperture of 0.22 μm.

Further conduct the utility model discloses technical scheme's improvement, protruding for being annular evenly distributed is in the three bump of body inner wall can reach the purpose of fixed milipore filter.

Further conduct the utility model discloses technical scheme's improvement, the recess is bellied cavity annular groove.

Further conduct the utility model discloses technical scheme's improvement, the salient with the recess does the inside integrative injection moulding of body to reinforce the structural strength of this part.

Further conduct the utility model discloses technical scheme's improvement, the tube cap with the top mouth of pipe of body adopts threaded connection or buckle to connect.

The beneficial effect that adopts this technical scheme to reach does:

1. through setting up the milipore filter into the obconic shape for the milipore filter is put to one side, and macromolecule deposit is in the milipore filter bottom during low-speed centrifugation, but does not block up milipore filter upper portion, has avoided prior art's milipore filter to place perpendicularly, and makes the problem that the milipore filter hole is blockked up by cell or biological macromolecule, thereby influences the exosome separation rate.

2. Through setting up several bumps at the body inner wall to and set up the recess in body bottom, make the connection between milipore filter and the body firm, guarantee that the milipore filter can not rock when the centrifugation and drop, cause and draw the failure, compare with prior art, a small amount of bumps are under the effect that reaches same fixed milipore filter, save material more, simple and convenient.

Drawings

Fig. 1 is a sectional view of the plane structure of the present invention.

1 pipe body, 2 ultrafiltration membranes, 3 salient points and 4 grooves

Detailed Description

For a better understanding of the technical solutions of the present technology, the present technology is described in detail below with reference to the accompanying drawings, and the description of the present technology is only exemplary and explanatory, and should not be construed as limiting the scope of the present technology in any way.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

It should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", and the like indicate orientations or positional relationships based on orientations or positional relationships shown in the drawings or orientations or positional relationships that are conventionally placed when the products of the art are used, and are used only for convenience in describing the technology and simplifying the description, but do not indicate or imply that the referred devices or elements must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the technology.

Furthermore, the terms "horizontal", "vertical", "overhang" and the like do not imply that the components are required to be absolutely horizontal or overhang, but may be slightly inclined. For example, "horizontal" merely means that the direction is more horizontal than "vertical" and does not mean that the structure must be perfectly horizontal, but may be slightly inclined.

In the description of the present technology, it should also be noted that, unless otherwise explicitly specified or limited, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meaning of the above terms in the present technology can be understood in a specific case to those of ordinary skill in the art.

As shown in fig. 1, the exosome extraction centrifuge tube comprises a tube body 1, a tube cover and an ultrafiltration membrane 2 in the tube body 1; the pore diameter of the ultrafiltration membrane 2 is between 0.15 and 0.30 mu m, the ultrafiltration membrane 2 is arranged in the tube body 1 in an inverted cone shape, and the top of the ultrafiltration membrane 2 is embedded into the groove 4 at the bottom of the tube body 1; the inner wall of the pipe body 1 is provided with a bulge 3 for fixing the upper edge of the ultrafiltration membrane 2, and when the centrifugal machine rotates, the ultrafiltration membrane 2 can be prevented from falling off.

Preferably, the ultrafiltration membrane 2 is an ultrafiltration membrane with a pore size of 0.22 μm.

Preferably, the protrusions 3 are three salient points 3 which are uniformly distributed on the inner wall of the tube body 1 in an annular shape, so that the purpose of fixing the ultrafiltration membrane 2 can be achieved.

Preferably, said groove 4 is a hollow annular groove 4 of the projection 3.

Preferably, the protruding points 3 and the grooves 4 are formed in the pipe body 1 through integral injection molding, so that the structural strength of the part is enhanced.

Preferably, the pipe cover is in threaded connection or buckled connection with the top end pipe orifice of the pipe body 1.

The utility model discloses a theory of use is: when the centrifugal tube is subjected to low-speed centrifugation, cells, cell debris and biomacromolecules can directly settle to the bottom of the ultrafiltration membrane 2, and the first step of an ultracentrifugation method is utilized; when the centrifugal speed is continuously increased, exosomes in the solution can directly pass through the ultrafiltration membrane 2 of 0.22um under the action of centrifugal force and enter the bottom of the tube body 1 of the centrifugal tube, and the step utilizes an ultrafiltration method; when the ultrafiltration membrane 2 is taken out, the required exosome extract is left in the body 1 of the centrifugal tube.

It should be noted that there are no specific structures in the above description, and it will be apparent to those skilled in the art that various modifications, decorations, or changes can be made without departing from the technical principles of the present invention; such modifications, variations, or combinations, or applying the concepts and solutions of the technology directly to other applications without further modifications, are intended to be within the scope of the present technology.

Claims

1. The utility model provides a exosome extraction centrifuging tube of combination ultrafiltration method and ultracentrifugation method which characterized in that: comprises a tube body, a tube cover and an ultrafiltration membrane in the tube body; the pore diameter of the ultrafiltration membrane is between 0.15 and 0.30 mu m, the ultrafiltration membrane is arranged in the tube body in an inverted cone shape, and the top of the ultrafiltration membrane is embedded into a groove at the bottom of the tube body; the inner wall of the pipe body is provided with a bulge for fixing the upper edge of the ultrafiltration membrane.

2. The combined ultrafiltration and ultracentrifugation exosome-extracting centrifuge tube of claim 1, wherein: the ultrafiltration membrane is an ultrafiltration membrane with the pore diameter of 0.22 mu m.

3. The combined ultrafiltration and ultracentrifugation exosome-extracting centrifuge tube of claim 1, wherein: the bulges are 2-5 salient points which are uniformly distributed on the inner wall of the tube body in an annular shape.

4. The combined ultrafiltration and ultracentrifugation exosome-extracting centrifuge tube of claim 1, wherein: the groove is a raised hollow annular groove.

5. The combined ultrafiltration and ultracentrifugation exosome-extracting centrifuge tube of claim 3, wherein: the convex points and the grooves are formed by integrally injection molding inside the pipe body.

6. The combined ultrafiltration and ultracentrifugation exosome-extracting centrifuge tube of claim 1, wherein: the pipe cover is connected with the top pipe orifice of the pipe body through threads or buckles.