Auxiliary device for exosome magnetic bead immunodetection

Abstract

The utility model discloses an exosome magnetic bead immunodetection auxiliary device, which relates to the technical field of medical scientific research assistance and comprises a shell part, a bearing part and an adjusting part, firstly, negative pressure acts on the outer side of the inner cylinder through a through hole of the outer cylinder by using a negative pressure assembly, the upper sides of the inner cylinder and the outer cylinder are sealed under the action of a blocking ring of an adjusting part, sugar, salt and small molecular protein of substances except exosomes put into the inner cylinder penetrate through an ultrafiltration membrane to leave the inner cylinder and the exosomes are left in the inner cylinder under the action of the ultrafiltration membrane on the lower side of the inner cylinder under the negative pressure, and then the exosomes are separated from the inner cylinder. An antigen-antibody binding buffer solution is added to bind exosomes with magnetic beads, an adjusting part is inverted, an electromagnetic assembly in a shell is started to attract the magnetic beads in an inner cylinder, a driving assembly is engaged with the inner cylinder, the inner cylinder rotates in an outer cylinder, and an upper-layer and lower-layer separation effect is formed on liquid in an inner cylinder pipe; the sealing plug seals the upper side of the inner cylinder while rotating along with the inner cylinder; a plurality of steps are completed in the inner cylinder through the change of the adjusting part.

Description

Technical Field

[0001] The utility model relates to the field of medical research auxiliary technology, and specifically to an auxiliary device for exosome magnetic bead immunoassay. Background Art

[0002] Exosomes are vesicles secreted by cells, with a diameter of generally 30-150nm. Exosomes carry a variety of proteins, biomarkers such as mRNA and miRNA, and are directly involved in important life activities such as information transfer between cells, antigen transmission, protein transport and RNA transcription. They are closely related to the occurrence of various diseases such as cancer, and therefore have great application value in the exploration of disease mechanisms and the detection of related diseases. However, exosomes usually exist in body fluids in a free form. The separation and purification of exosomes is the basis for the application of disease mechanisms and disease detection based on exosomes. In recent years, researchers have developed a variety of methods and technologies for the separation and purification of exosomes using the biophysical and biochemical properties of exosomes, mainly ultracentrifugation, polymer precipitation, immunoseparation, and microfluidics-based separation.

[0003] Immunomagnetic bead (IMB) technology is a technical method that appeared in the 1980s. Based on immunology, it has penetrated into various fields such as pathology, physiology, pharmacology, microbiology, biochemistry and molecular genetics. It is increasingly used in immunoassays, cell separation, biomacromolecule purification and molecular biology.

[0004] In summary, the current exosome separation process has the following technical problems: exosomes are often separated by the immunomagnetic bead method, where immunoreactive antibodies are coated on the surface of the magnetic beads for antigen-antibody reaction, and the cells bound to the magnetic beads are placed under a magnetic field to distinguish them from other unbound cells, thereby completing the screening or removing of the marked cells, thereby achieving the purpose of positive or negative selection of cells; small molecules, such as sugars, salts, and small molecules of proteins, need to be removed from the sample first; in the prior art, due to the variety of processes, the method of separating exosomes is complicated and takes a long time, and it is necessary to design a multifunctional device to solve the above problems. Utility Model Content

[0005] In view of the above problems existing in the prior art, the utility model provides an auxiliary device for exosome magnetic bead immunoassay.

[0006] In order to achieve the above technical objectives and the above technical effects, the present invention is implemented through the following technical solutions:

[0007] An auxiliary device for exosome magnetic bead immunoassay comprises a shell part, a bearing part and an adjustment part, wherein the shell part comprises an outer cylinder and a negative pressure assembly, the bearing part comprises an inner cylinder and an upper gear, and the adjustment part comprises a blocking ring and a driving assembly; the inner cylinder is rotatably connected inside the outer cylinder, the outer cylinder is connected to the negative pressure assembly, the lower end of the inner cylinder is connected to an ultrafiltration membrane, the upper end of the inner cylinder is fixedly connected to the upper gear, the upper sides of the inner cylinder and the outer cylinder are provided with an adjustment part, the blocking ring is slidably connected between the outer cylinder and the inner cylinder, the adjustment part is provided with a driving assembly on the other side of the blocking ring, the driving assembly is meshedly connected to the upper gear, and only one of the blocking ring and the driving assembly of the adjustment part can be selected to work.

[0008] Furthermore, the adjustment part also includes an upper cover and a ring frame, the driving assembly includes a motor and a middle gear, the upper cover is fixedly connected to a sealing ring, the position where the outer shell is connected to the sealing ring and the inner cylinder are provided with corresponding holes, the upper cover is fixedly connected to the ring frame on the other side of the sealing ring, the upper cover is connected to the driving assembly on one side of the ring frame, the motor is fixedly connected in the middle of the upper cover, the motor is connected to the middle gear, and the middle gear is meshingly connected to the upper gear.

[0009] Furthermore, a connecting frame is connected inside the ring frame, and a sealing plug is rotatably connected to the connecting frame. The sealing plug is made of rubber and is detachably connected to the port of the inner cylinder. The sealing plug and the middle gear are at the same horizontal position.

[0010] Furthermore, the sealing plug is connected to the connecting frame via an upper bearing; and a lower bearing is connected between the inner cylinder and the outer cylinder.

[0011] Furthermore, the outer cylinder is provided with four groups, the outer cylinder is fixedly connected to the outer shell, the negative pressure assembly includes a connecting pipe, a connecting pipe and a negative pressure pump, the four groups of outer cylinders are fixedly connected to the connecting pipes, the outer cylinder is provided with a through hole at the connection position with the connecting pipe, the connecting pipe is fixedly connected to the lower side of the connecting pipe, the connecting pipe is fixedly connected to the negative pressure pump on the outside of the outer shell, and the negative pressure pump is provided with an indicator dial displaying the negative pressure value.

[0012] Furthermore, the ultrafiltration membrane is a 100KDa ultrafiltration membrane.

[0013] Furthermore, the negative pressure component has a working negative pressure of -10 to -30 KPa.

[0014] Furthermore, the housing is provided with an electromagnetic component.

[0015] Beneficial effects of the utility model:

[0016] An auxiliary device for exosome magnetic bead immunoassay, firstly uses a negative pressure component to apply negative pressure to the outside of an inner cylinder through the through hole of an outer cylinder, under the action of a sealing ring of an adjustment part, the inner cylinder and the upper side of an outer cylinder are sealed, and under the action of an ultrafiltration membrane on the lower side of the inner cylinder, the negative pressure causes the substances placed in the inner cylinder, except sugar, salt, and small molecule protein of the exosome, to leave the inner cylinder through the ultrafiltration membrane, and the exosomes are retained in the inner cylinder for subsequent work such as adding a cleaning solution, adding an antigen-antibody binding buffer to combine the exosomes with the magnetic beads, inverting the adjustment part, starting the electromagnetic component in the outer shell to attract the magnetic beads in the inner cylinder, the driving component meshes with the inner cylinder, and the inner cylinder rotates in the outer cylinder, forming an upper and lower layer separation effect of the liquid inside the inner cylinder tube, and the sealing plug seals the upper side of the inner cylinder while following the rotation of the inner cylinder; multiple steps are completed in the inner cylinder by changing the adjustment part.

[0017] Of course, any product implementing the present invention does not necessarily need to achieve all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the utility model, the drawings required for describing the embodiments are briefly introduced below. Obviously, the drawings described below are only some embodiments of the utility model. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0019] Figure 1 This is a schematic diagram of the overall structure of the auxiliary device for exosome magnetic bead immunoassay according to an embodiment of the utility model;

[0020] Figure 2 This is a schematic diagram of the structural connection between the outer shell and the outer cylinder according to an embodiment of the utility model;

[0021] Figure 3 This is a schematic diagram of the structure of the outer cylinder, the connecting pipe and the connecting pipe according to the embodiment of the utility model;

[0022] Figure 4 This is a schematic diagram of the structure of the inner cylinder and the negative pressure assembly according to an embodiment of the utility model;

[0023] Figure 5 This is a schematic diagram of the structure of the inner cylinder described in the embodiment of the utility model;

[0024] Figure 6 This is a schematic diagram of the lower structure of the inner cylinder according to an embodiment of the utility model;

[0025] Figure 7 This is a schematic diagram of the cross-sectional structure connection of the outer shell and the outer cylinder according to the embodiment of the utility model;

[0026] Figure 8It is a structural schematic diagram of the adjustment part according to an embodiment of the utility model;

[0027] Fig. 9 It is a schematic diagram of the cross-sectional structure of the adjustment part according to the embodiment of the utility model;

[0028] Fig.10 It is a schematic diagram of the cross-sectional structure connection of the adjustment part, the outer shell and the outer cylinder according to the embodiment of the utility model;

[0029] Fig.11 It is a schematic diagram of the cross-sectional structural connection of the outer shell, the outer cylinder and the inverted adjustment part described in the embodiment of the utility model.

[0030] In the accompanying drawings, the components represented by the reference numerals are listed as follows:

[0031] 1-housing, 101-outer cylinder, 102-through hole, 103-connecting pipe, 104-connecting pipe, 105-negative pressure pump, 106-indicator plate, 2-inner cylinder, 201-upper gear, 202-lower bearing, 203-attraction membrane, 3-upper cover, 301-sealing ring, 310-ring frame, 311-motor, 312-middle gear, 320-connecting frame, 321-sealing plug, 322-upper bearing. DETAILED DESCRIPTION

[0032] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0033] Example

[0034] like Figure 1-11 Shown

[0035] The utility model discloses an auxiliary device for exosome magnetic bead immunoassay, comprising a shell part, a bearing part and an adjustment part, wherein the shell part comprises an outer cylinder (101) and a negative pressure component, the bearing part comprises an inner cylinder (2) and an upper gear (201), and the adjustment part comprises a blocking ring (301) and a driving component; the inner cylinder (2) is rotatably connected inside the outer cylinder (101), the outer cylinder (101) is connected to the negative pressure component, the lower end of the inner cylinder (2) is connected to an ultrafiltration membrane, the upper end of the inner cylinder (2) is fixedly connected to the upper gear (201), the upper sides of the inner cylinder (2) and the outer cylinder (101) are provided with an adjustment part, the blocking ring (301) is slidably connected between the outer cylinder (101) and the inner cylinder (2), the adjustment part is provided with a driving component on the other side of the blocking ring (301), the driving component is meshedly connected to the upper gear (201), and the blocking ring (301) and the driving component of the adjustment part can only select one of them to work;

[0036] The adjustment portion further comprises an upper cover (3) and a ring frame (310); the driving assembly comprises a motor (311) and a middle gear (312); a blocking ring (301) is fixedly connected to the upper cover (3); a corresponding hole is opened in the inner cylinder (2) at a position where the outer shell (1) is connected to the blocking ring (301); the upper cover (3) is fixedly connected to the ring frame (310) on the other side of the blocking ring (301); the upper cover (3) is connected to the driving assembly on one side of the ring frame (310); the motor (311) is fixedly connected to the middle of the upper cover (3); the motor (311) is connected to the middle gear (312); and the middle gear (312) is meshedly connected to the upper gear (201);

[0037] The ring frame (310) is also connected to a connecting frame (320), and a sealing plug (321) is rotatably connected to the connecting frame (320). The sealing plug (321) is made of rubber material, and is detachably connected to the port of the inner cylinder (2). The sealing plug (321) and the middle gear (312) are at the same horizontal position; the sealing plug (321) can prevent substances from flowing out of the inner cylinder during subsequent centrifugal rotation.

[0038] The sealing plug (321) is connected to the connecting frame (320) via an upper bearing (322); a lower bearing (202) is connected between the inner cylinder (2) and the outer cylinder (101);

[0039] The outer cylinder (101) is provided with four groups, the outer cylinder (101) is fixedly connected to the outer shell (1), the negative pressure assembly comprises a connecting pipe (103), a connecting pipe (104) and a negative pressure pump (105), the four groups of the outer cylinders (101) are fixedly connected to the connecting pipe (103), the outer cylinder (101) is provided with a through hole (102) at a position where it is connected to the connecting pipe (103), the lower side of the connecting pipe (103) is fixedly connected to the connecting pipe (104), the connecting pipe (104) is fixedly connected to the negative pressure pump (105) on the outer side of the outer shell (1), and the negative pressure pump (105) is provided with an indicator plate (106) for displaying the negative pressure value;

[0040] The ultrafiltration membrane is a 100KDa ultrafiltration membrane; the negative pressure component has a working negative pressure of -10 to -30KPa; and the housing (1) is provided with an electromagnetic component.

[0041] In summary, the utility model is an auxiliary device for exosome magnetic bead immunoassay, which first uses a negative pressure component to apply negative pressure to the outside of the inner cylinder through the through hole of the outer cylinder, and under the action of the blocking ring of the adjustment part, the inner cylinder and the upper side of the outer cylinder are sealed, and under the action of the ultrafiltration membrane on the lower side of the inner cylinder, the negative pressure allows the substances placed in the inner cylinder, except the sugar, salt, and small molecule protein of the exosome, to leave the inner cylinder through the ultrafiltration membrane, and the exosomes are retained in the inner cylinder for subsequent work such as adding cleaning liquid, adding antigen-antibody binding buffer to combine the exosomes with the magnetic beads, inverting the adjustment part, starting the electromagnetic component in the outer shell to attract the magnetic beads in the inner cylinder, the driving component engages with the inner cylinder, and the inner cylinder rotates in the outer cylinder to separate the liquid inside the inner cylinder into two layers, and the sealing plug blocks the upper side of the inner cylinder while following the rotation of the inner cylinder; multiple steps are completed in the inner cylinder by changing the adjustment part, and after separation, the upper and lower layers of liquid are extracted respectively and in turn, completing the auxiliary operation in the process of exosome magnetic bead immunoassay.

[0042] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the utility model. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0043] The preferred embodiments of the utility model disclosed above are only used to help explain the utility model. The preferred embodiments do not describe all the details in detail, nor do they limit the utility model to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the utility model, so that technicians in the relevant technical field can well understand and use the utility model. The utility model is limited only by the claims and their full scope and equivalents.

Claims

1. An auxiliary device for exosome magnetic bead immunoassay, characterized in that: It comprises a shell part, a bearing part and an adjustment part, the shell part comprises an outer cylinder and a negative pressure assembly, the bearing part comprises an inner cylinder and an upper gear, and the adjustment part comprises a blocking ring and a driving assembly; the inner cylinder is rotatably connected in the outer cylinder, the outer cylinder is connected to the negative pressure assembly, the lower end of the inner cylinder is connected to an ultrafiltration membrane, the upper end of the inner cylinder is fixedly connected to the upper gear, the upper sides of the inner cylinder and the outer cylinder are provided with an adjustment part, the blocking ring is slidably connected between the outer cylinder and the inner cylinder, the adjustment part is provided with a driving assembly on the other side of the blocking ring, the driving assembly is meshed and connected with the upper gear, and the blocking ring and the driving assembly of the adjustment part can only select one of them to work; The outer shell is fixedly connected to the outer cylinder.

2. The auxiliary device for exosome magnetic bead immunoassay according to claim 1, characterized in that: The adjustment part also includes an upper cover and a ring frame, the driving assembly includes a motor and a middle gear, a sealing ring is fixedly connected to the upper cover, a corresponding hole is opened in the position where the outer shell is connected to the sealing ring and the inner cylinder, the upper cover is fixedly connected to the ring frame on the other side of the sealing ring, the upper cover is connected to the driving assembly on one side of the ring frame, the motor is fixedly connected in the middle of the upper cover, the motor is connected to the middle gear, and the middle gear is meshed with the upper gear.

3. The auxiliary device for exosome magnetic bead immunoassay according to claim 2, characterized in that: The ring frame is also connected to a connecting frame, and a sealing plug is rotatably connected to the connecting frame. The sealing plug is made of rubber and is detachably connected to the port of the inner cylinder. The sealing plug and the middle gear are at the same horizontal position.

4. The auxiliary device for exosome magnetic bead immunoassay according to claim 3, characterized in that: The sealing plug is connected to the connecting frame through an upper bearing; a lower bearing is connected between the inner cylinder and the outer cylinder.

5. The auxiliary device for exosome magnetic bead immunoassay according to claim 1, characterized in that: The outer tubes are provided with four groups, and the negative pressure assembly includes a connecting tube, a connecting tube and a negative pressure pump. The four groups of outer tubes are fixedly connected with the connecting tubes. The outer tubes are provided with through holes at the connection positions with the connecting tubes. The connecting tubes are fixedly connected with connecting tubes on the lower sides. The connecting tubes are fixedly connected with the negative pressure pump on the outer side of the outer shell. The negative pressure pump is provided with an indicator dial for displaying the negative pressure value.

6. The auxiliary device for exosome magnetic bead immunoassay according to claim 1, characterized in that: The ultrafiltration membrane is a 100KDa ultrafiltration membrane.

7. The auxiliary device for exosome magnetic bead immunoassay according to claim 1, characterized in that: The negative pressure component has a working negative pressure of -10 to -30 KPa.

8. The auxiliary device for exosome magnetic bead immunoassay according to claim 1, characterized in that: The housing is provided with an electromagnetic component.

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