WO2020073829A1 - Dispositif de séparation et son utilisation, système de détection, système de détection électrochimique et système de tri cellulaire - Google Patents
Dispositif de séparation et son utilisation, système de détection, système de détection électrochimique et système de tri cellulaire Download PDFInfo
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- WO2020073829A1 WO2020073829A1 PCT/CN2019/108677 CN2019108677W WO2020073829A1 WO 2020073829 A1 WO2020073829 A1 WO 2020073829A1 CN 2019108677 W CN2019108677 W CN 2019108677W WO 2020073829 A1 WO2020073829 A1 WO 2020073829A1
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- Prior art keywords
- chamber
- separation device
- functional
- detection system
- sample
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- 238000000926 separation method Methods 0.000 title claims abstract description 123
- 238000001514 detection method Methods 0.000 title claims abstract description 78
- 238000000835 electrochemical detection Methods 0.000 title claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 63
- 230000000903 blocking effect Effects 0.000 claims abstract description 19
- 238000001179 sorption measurement Methods 0.000 claims description 52
- 239000002699 waste material Substances 0.000 claims description 50
- 238000004140 cleaning Methods 0.000 claims description 30
- 230000007246 mechanism Effects 0.000 claims description 24
- 239000012528 membrane Substances 0.000 claims description 24
- 239000011324 bead Substances 0.000 claims description 23
- 238000002038 chemiluminescence detection Methods 0.000 claims description 19
- 238000006243 chemical reaction Methods 0.000 claims description 15
- 230000009471 action Effects 0.000 claims description 13
- 238000007789 sealing Methods 0.000 claims description 13
- 102000039446 nucleic acids Human genes 0.000 claims description 8
- 108020004707 nucleic acids Proteins 0.000 claims description 8
- 150000007523 nucleic acids Chemical class 0.000 claims description 8
- 238000000605 extraction Methods 0.000 claims description 5
- 238000005070 sampling Methods 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 229910052814 silicon oxide Inorganic materials 0.000 claims description 3
- 238000011109 contamination Methods 0.000 abstract description 5
- 238000005192 partition Methods 0.000 abstract description 4
- 239000013076 target substance Substances 0.000 description 62
- 210000004027 cell Anatomy 0.000 description 34
- 239000003463 adsorbent Substances 0.000 description 23
- 239000003153 chemical reaction reagent Substances 0.000 description 19
- 239000000243 solution Substances 0.000 description 15
- 238000003752 polymerase chain reaction Methods 0.000 description 12
- 239000006166 lysate Substances 0.000 description 9
- 239000007795 chemical reaction product Substances 0.000 description 7
- 238000000034 method Methods 0.000 description 6
- 238000002156 mixing Methods 0.000 description 5
- -1 polyethylene Polymers 0.000 description 4
- 230000009089 cytolysis Effects 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 239000004743 Polypropylene Substances 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 239000004809 Teflon Substances 0.000 description 2
- 229920006362 Teflon® Polymers 0.000 description 2
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 2
- 239000012295 chemical reaction liquid Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000009169 immunotherapy Methods 0.000 description 2
- 238000009434 installation Methods 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 238000004020 luminiscence type Methods 0.000 description 2
- 239000000696 magnetic material Substances 0.000 description 2
- 229920006284 nylon film Polymers 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 229920000098 polyolefin Polymers 0.000 description 2
- 229920001155 polypropylene Polymers 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000004451 qualitative analysis Methods 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 238000001069 Raman spectroscopy Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000002826 magnetic-activated cell sorting Methods 0.000 description 1
- 230000005389 magnetism Effects 0.000 description 1
- 239000011325 microbead Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000013077 target material Substances 0.000 description 1
- 238000003466 welding Methods 0.000 description 1
Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/06—Magnetic means
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/101—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
Definitions
- the present application relates to the technical field of separation devices, in particular to a separation device and its application, detection system, electrochemical detection system and cell sorting system.
- the magnetic bead separation technology mainly uses magnetic beads to specifically adsorb the material to be separated to separate the material to be separated. Because of its strong specificity, safety and non-toxicity, it is widely used in nucleic acid extraction and chemiluminescence detection. At present, during the use of most magnetic bead separation devices, the sample to be tested needs to be loaded into a centrifuge tube and then placed in the magnetic bead separation device for separation operation. This method requires the process of repeatedly opening and adding the liquid, mixing and mixing the cover, and opening and discarding the liquid. Repeated opening of the cover is likely to cause contamination of the sample to be tested.
- a separation device including:
- the barrel has opposite first and second ends;
- the diaphragm assembly includes a plurality of separation membranes and a plurality of sealing membranes.
- the plurality of separation membranes are all accommodated in the cylinder body and are all sealedly connected to the cylinder body.
- the plurality of separation membranes extend from the second end Spaced toward the first end to divide the barrel into an end chamber and N functional chambers, the end chamber is close to the second end, the sidewalls of the N functional chambers There are drain holes on the openings, and each of the drain holes is blocked with the blocking film; and
- the slider is provided near the first end and can slide toward the second end.
- the cylinder body is divided into an end chamber and N functional chambers by a plurality of separation membranes, and the plurality of separation membranes are all sealedly connected to the cylinder body, and the side walls of the N functional chambers are all provided with Drainage holes, each of which is sealed with a blocking membrane, can be placed in the functional chamber closest to the first end to be configured to adsorb the target material adsorbent, can be placed in other functional chambers or the reaction liquid Cleaning fluid, etc .; because the slider is located near the first end and can slide toward the second end, when the target substance needs to be separated, the slider can be accommodated in the functional chamber closest to the first end and can be slid An adsorption member is provided on the side away from the first end, and the sample to be separated is added to adsorb the target substance on the adsorption member, and the sliding member is slid toward the second end, so that the sealing membrane and the separation membrane are broken and carried The adsorbent that adsorbs the target
- the separating device further includes a pushing member, and the pushing member is detachably connected with the sliding member.
- it further includes an adsorption member, which is accommodated in the functional chamber closest to the first end, the pushing member is at least partially magnetic, and when the pushing member is connected to the sliding member The pushing member can adsorb the adsorption member and the adsorption member can adsorb on the sliding member.
- an adsorption member which is accommodated in the functional chamber closest to the first end, the pushing member is at least partially magnetic, and when the pushing member is connected to the sliding member The pushing member can adsorb the adsorption member and the adsorption member can adsorb on the sliding member.
- the separation device further includes a magnetic field generating mechanism configured to provide a magnetic field to the adsorption member, and the adsorption member can move under the action of the magnetic field.
- a magnetic field generating mechanism configured to provide a magnetic field to the adsorption member, and the adsorption member can move under the action of the magnetic field.
- the adsorption member is a magnetic bead.
- the magnetic beads are superparamagnetic silicon oxide nano magnetic beads or carboxylated magnetic beads.
- an adsorption member is further included, and the adsorption member is a chip.
- the adsorption member is accommodated in the functional chamber closest to the first end and fixed to the sliding member.
- the tensile strength of the separation film is greater than the tensile strength of the blocking film, and when the sliding member slides in a direction close to the second end, the functional cavity where the sliding member is located can be made The pressure in the chamber is increased, so that the blocking film and the separation film of the functional chamber where the slider is located can be sequentially broken.
- a waste liquid tank is further included, and the waste liquid tank is in communication with the drain hole.
- the waste liquid tank is a ring-shaped hollow structure
- the cylinder is at least partially contained in the waste liquid tank, and is sealedly connected to the waste liquid tank;
- the waste liquid tank is cylindrical, and the outer wall of the waste liquid tank is fixed to the outer wall of the barrel body.
- a side wall of the adsorption chamber is further provided with an air pressure balancing hole communicating with the waste liquid tank, and the air pressure balancing hole is provided near the first end.
- the cylinder body is provided with a sample port, the sample port is located on the cavity wall of the functional chamber closest to the first end, and the separation device further includes a configuration configured to seal the sample port Sample port seals;
- the cylinder body is provided with a sample outlet, the sample outlet is communicated with the end chamber, and the separation device further includes a stopper configured to seal the sample outlet.
- the barrel includes a barrel body, a first side plate and a second side plate;
- the first end and the second end are provided with a first opening and a second opening, the first opening is opposite to the second opening, and the first side plate and the second side plate respectively shield the The first opening and the second opening are both hermetically connected to the barrel body.
- the sliding member includes a sliding part and a mounting part fixedly connected to the sliding part;
- the sliding portion is provided near the first end and can slide toward the second end; the sliding portion is housed in the functional chamber near the first end and is connected to the body of the cylinder Sealed connection on the inner wall;
- the mounting portion is provided near the first end and can slide toward the second end; the mounting portion is partially accommodated in the functional chamber near the first end and can seal the cylinder.
- a detection system includes the separation device described above.
- the detection system is a PCR (Polymerase Chain Reaction) detection system
- the end chamber is a detection chamber
- the functional chamber closest to the first end is a sample chamber
- At least one of the remaining functional chambers is a cleaning chamber
- the detection system is a chemiluminescence detection system
- the end chamber is a detection chamber
- the functional chamber closest to the first end is a sample chamber
- An electrochemical detection system is characterized by including the above-mentioned separation device.
- the end chamber is a detection chamber
- the functional chamber closest to the first end is a sample chamber
- at least one of the remaining functional chambers is a cleaning chamber.
- a cell sorting system is characterized by including the above-mentioned separation device.
- FIG. 1 is a schematic structural diagram of a separation device according to an embodiment
- FIG. 2 is a schematic structural view of the separation device shown in FIG. 1 after omitting the waste liquid tank;
- FIG. 3 is a schematic structural view of a sliding member and a pushing member in the separation device shown in FIG. 1;
- FIG. 4 is a schematic structural view of a separation device in a chemiluminescence detection system after a waste liquid tank is omitted.
- Icons 10-separation device; 100-barrel; 110-barrel body; 111-first end; 113-second end; 120-first side plate; 130-second side plate; 140-end chamber 144-air pressure balance hole; 150-function chamber; 160-drain hole; 170-sample inlet; 190-sample outlet; 200-diaphragm assembly; 210-partition membrane; 220-blocking membrane; 300-slide Parts; 310-sliding part; 320-mounting part; 322-accommodating groove; 400-pushing part; 500-seal; 600-waste liquid tank.
- the detection system of an embodiment includes a separation device 10 and a detection device (not shown).
- the separation device 10 is configured to separate the target substance in the sample to be measured.
- the detection device communicates with the separation device 10 to perform qualitative or quantitative detection of the target substance in the sample to be tested.
- the sample to be tested is blood or saliva.
- the target substances are nucleic acids and target cells. It should be noted that the sample to be tested is not limited to the above-mentioned pointed sample, and the target substance is not limited to the above-mentioned pointed substance, and can be set according to the actual situation.
- the separation device 10 includes a cylinder 100, a diaphragm assembly 200, a sliding member 300, a pushing member 400, and a waste tank 600.
- the target substance is a nucleic acid.
- the separation device 10 is configured for nucleic acid extraction.
- the barrel 100 is the outer shell of the separating device 10.
- the barrel 100 includes a barrel body 110, a first side plate 120 and a second side plate 130.
- the barrel body 110 has a first end 111 and a second end 113 opposite to each other.
- the first end 111 and the second end 113 define a first opening (not shown) and a second opening (not shown).
- the first opening is opposite to the second opening.
- the first side plate 120 and the second side plate 130 respectively cover the first opening and the second opening, and are both sealedly connected to the barrel body 110.
- cylindrical body 110, the second side plate 130, and the first side plate 120 may be an integrally formed structure, or may be other fixing methods, such as welding.
- the cylindrical body 110 is cylindrical. Both the first side plate 120 and the second side plate 130 are disc-shaped. The first side plate 120 is opposite to and parallel to the second side plate 130. The outer diameter of the first side plate 120 is approximately the same as the inner diameter of the first opening, so that the first side plate 120 just covers the first opening; The second side plate 130 just covers the second opening.
- cylindrical body 110 is not limited to the above-mentioned pointed shape, and may be other shapes, such as a square cylindrical shape.
- first side plate 120 and the second side plate 130 are not limited to the above-mentioned shapes, and may be other shapes, such as a square shape. It can be set according to the actual situation, as long as the first side plate 120 and the second side plate 130 cover the first opening and the second opening, respectively, and both are fixedly connected and hermetically connected to the barrel body 110.
- the diaphragm assembly 200 includes a plurality of separation films 210 and a plurality of blocking films 220.
- the plurality of separation films 210 are all contained in the cylinder 100 and are all sealedly connected to the cylinder 100.
- the separation membranes 210 are arranged at intervals from the second end 113 to the first end 111 to divide the cylinder 100 into an end chamber 140 and N functional chambers 150.
- the end chamber 140 is disposed near the second end 113.
- the side walls of the N functional chambers 150 are all provided with drainage holes 160, and each drainage hole 160 is sealed with a blocking film 220.
- the detection system is a PCR (Polymerase Chain Reaction) detection system.
- the end chamber 140 is a detection chamber
- the functional chamber 150 closest to the first end is a sample chamber
- at least one of the remaining functional chambers 150 is a cleaning chamber.
- each separation film 210 has a disc shape, and is arranged parallel to the first side plate 120.
- the three separation films 210 are arranged at intervals from the second end 113 to the first end 111 to divide the cylinder 100 into an end chamber 140 and three functional chambers 150.
- the end chamber 140 is configured to contain the eluent.
- the functional chamber 150 closest to the first end 111 can accommodate the sample to be tested.
- the remaining two functional chambers 150 are configured to contain cleaning fluid to clean the target substance.
- the side walls of the three functional chambers 150 are all provided with drainage holes 160.
- the three blocking films 220 respectively block the drainage holes 160 of the three functional chambers 150.
- the cleaning liquids in the two functional chambers 150 configured to receive the cleaning liquid may be the same cleaning liquid or different cleaning liquids, and can be set according to actual conditions.
- the separation film 210 is an acrylic film, a nylon film, a Teflon film, a polyethylene film, a polypropylene film, or a polyolefin film. It should be noted that the separation film 210 is not limited to the above-mentioned film, and any film commonly used in the art may be used as the separation film 210.
- the blocking film 220 is an acrylic film, nylon film, Teflon film, polyethylene film, polypropylene film or polyolefin film. It should be noted that the blocking film 220 is not limited to the above-mentioned film, and any film commonly used in the art may be used as the blocking film 220.
- the cylinder 100 is provided with a sample port 170.
- the sample port 170 is located on the wall of the functional chamber 150 closest to the first end 111.
- the sample to be tested can be added into the barrel 100 from the sample port 170.
- the separation device 10 further includes a sealing member 500 configured to seal the sampling port 170.
- the sample port 170 is spaced from and opposite to the drain hole 160.
- the sealing member 500 is a plug and can be inserted into the sample port 170 to seal the sample port 170.
- the sealer 500 is not limited to a plug, but may also be a lid body, which is provided on the sampling port 170 to seal the sampling port 170.
- the cylinder 100 is provided with a sample outlet 190.
- the sample outlet 190 communicates with the end chamber 140.
- the separation device 10 further includes a stopper (not shown) configured to seal the sample outlet 190.
- the sample outlet 190 is opened in the second side plate 130 and penetrates the second side plate 130.
- the occluder may be a plug so as to be able to be inserted into the sample outlet 190 and seal the sample outlet 190.
- the stopper may also be a cover so as to cover the sample outlet 190 and seal the sample outlet 190.
- the sample port 170 and the sample port 190 can be set as a circular through hole or a square through hole, as long as the sample to be tested can be added to the barrel 100 from the sample port 170, and the tested sample can be taken out
- the sample port 190 only needs to exit the barrel 100, and will not be repeated here.
- the slider 300 is disposed near the first end 111 and can slide toward the second end 113. Further, the tensile strength of the separation film 210 is greater than the tensile strength of the blocking film 220.
- the pressure in the functional chamber 150 where the slider 300 is located can be increased, and the sealing film 220 and the separation membrane 210 of the functional chamber 150 where the slider 300 is located can be increased. Able to break in sequence.
- the slider 300 is at least partially accommodated in the functional chamber 150 closest to the first end 111 and seals the functional chamber 150 closest to the first end 111.
- the sliding member 300 includes a sliding part 310 and a mounting part 320 fixed to the sliding part 310.
- the sliding portion 310 is provided near the first end 111 and can slide toward the second end 113. Further, the sliding portion 310 is accommodated in the functional chamber 150 closest to the first end 111 and is sealingly connected to the inner wall of the barrel body 110. In the illustrated embodiment, the sliding portion 310 has a disc shape. The sliding part 310 is provided in parallel with the separation film 210.
- the mounting portion 320 is provided near the first end 111 and can slide toward the second end 113. Further, the mounting portion 320 is partially accommodated in the functional chamber 150 closest to the first end 111 and seals the cylindrical body 100. In the illustrated embodiment, the mounting portion 320 is substantially cylindrical. The mounting portion 320 is located on the side of the sliding portion 310 close to the first side plate 120. One end of the mounting part 320 is fixed to the sliding part 310.
- the cylinder 100 is provided with a mounting hole (not shown) communicating with the functional chamber 150 closest to the first end 111.
- the slider 300 can penetrate the mounting hole and be at least partially received in the functional chamber 150 closest to the first end 111.
- the mounting hole is opened on the first side plate 120.
- the mounting portion 320 penetrates the mounting hole and is partially accommodated in the functional chamber 150 closest to the first end 111.
- the pushing member 400 and the sliding member 300 are detachably connected. Further, the pushing member 400 can slide against the pushing member 300.
- the pushing member 400 has a rod shape.
- the mounting portion 320 defines a receiving slot 322.
- the receiving slot 322 is opened at the end of the mounting part 320 away from the sliding part 310.
- the pushing member 400 can be at least partially received in the receiving groove 322 to slide against the pushing member 300.
- the accommodating groove 322 is formed inward by the end of the mounting portion 320 away from the sliding portion 310.
- the separation device 10 further includes an adsorption member (not shown).
- the suction member is accommodated in the functional chamber 150 closest to the first end 111.
- the pushing member 400 is at least partially magnetic. When the pushing member 400 is connected to the sliding member 300, the pushing member 400 can provide a magnetic field to the adsorption member, and fix the adsorption member on the sliding member 300.
- the suction member is accommodated in the functional chamber 150 closest to the first end 111 and located on the side of the sliding part 310 away from the mounting part 320.
- the side of the pushing member 400 close to the slider 300 has magnetism to provide a magnetic field to the suction member so that the suction member can be fixed on the side of the sliding part 310 away from the mounting part 320.
- the pushing member 400 may also be directly made of magnetic material.
- the adsorption member is a magnetic bead.
- the magnetic beads are superparamagnetic silicon oxide nano magnetic beads or carboxylated magnetic beads. It should be noted that the adsorbent is not limited to the magnetic beads indicated above, but can also be other magnetic beads, which can be set according to the actual situation, as long as it can be combined with the target substance in the sample to be tested.
- the separation device 10 further includes a lysis solution to lyse the sample to be tested to release the target substance.
- the lysate is contained in the functional chamber 150 closest to the first end 111.
- the lysate is a cell lysate or a protein lysate. It should be noted that the lysate is not limited to the lysate indicated above, but can also be other lysates, which can be set according to the actual situation, as long as the target substance in the sample to be tested can be released.
- the separation device 10 further includes a magnetic field generating mechanism (not shown) configured to provide a magnetic field to the adsorption member.
- the adsorbent can move under the action of the magnetic field. By adding an external magnetic field, the adsorbent can be moved to achieve the effect of mixing.
- the magnetic field generating mechanism may be a magnetic field generating mechanism with an electromagnetic coil, or a magnetic field generating mechanism with a permanent magnet.
- the waste liquid tank 600 communicates with the liquid discharge hole 160. Further, the waste liquid tank 600 is cylindrical. The outer wall of the waste liquid tank 600 is fixed to the outer wall of the cylinder 100. In the illustrated embodiment, the extending direction of the waste liquid tank 600 is substantially parallel to the extending direction of the cylinder 100. An installation port (not shown) is opened on the outer peripheral surface of the waste liquid tank 600. The outer wall of the waste liquid tank 600 is fixed to the outer wall of the cylinder 100 and shields the installation opening, so that the waste liquid tank 600 and the cylinder 100 cooperate to form a receiving chamber (not shown).
- the side wall of the cylinder 100 is further provided with an air pressure balancing hole 144 communicating with the waste liquid tank 600.
- the air pressure balance hole 144 is disposed near the first end 111.
- the air pressure balancing hole 144 is configured to balance the air pressure in the cylinder 100.
- the detection device communicates with the separation device 10 to perform qualitative or quantitative detection of the target substance in the sample to be tested.
- the detection device is a PCR (polymerase chain reaction) detection device.
- the detection device is a microfluidic PCR (polymerase chain reaction) detection device.
- the detection device can be hermetically connected to the sample outlet 190 so that the target substance in the end chamber 140 can enter the detection device for detection.
- the operation process of the above separation device 10 is as follows:
- the sliding part 310 is accommodated in the functional chamber 150 closest to the first end 111, and the sliding part 310 is positioned between the sample port 170 and the air pressure balance hole 144, An adsorption member is added to the functional chamber 150 closest to the first end 111, and the adsorption member is located on the side of the sliding part 310 away from the mounting part 320, and a cleaning solution is added to the other functional chamber 150, and the end chamber 140 Add eluent. It should be noted that, if the sample to be tested needs to be cracked to release the target substance to be adsorbed on the adsorption member, the lysis solution is added to the functional chamber 150 closest to the first end 111.
- the sample to be tested is added into the functional chamber 150 closest to the first end 111 from the sample loading port 170, and the magnetic field generating mechanism is turned on to provide a magnetic field to the barrel 100, so that the sample to be tested and the adsorbent are mixed evenly. If a lysate is added, the sample to be tested is decomposed under the action of the lysate to release the target substance, and the released target substance is adsorbed with the adsorbent to obtain an adsorbent carrying the target substance.
- the pushing member 400 is received in the receiving groove 322 to push the sliding member 300 and fix the suction member.
- the sliding member 300 moves toward the second end 113 by the pushing member 400, and the sealing film 220 and the separating film 210 of the functional chamber 150 where the sliding part 310 is located are sequentially broken under the action of the air pressure, and the waste liquid is discharged from the
- the liquid hole 160 flows into the waste liquid tank 600, and the sliding member 300 continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150, and stops moving, so that the pushing member 400 moves toward the first end 111 and cannot Fix the suction element.
- the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100, so that the adsorbent and the cleaning solution are mixed to clean the adsorbent carrying the target substance.
- the pushing member 400 is received in the receiving groove 322 to push the sliding member 300 and fix the suction member.
- the sliding member 300 moves toward the second end 113 by the pushing member 400, and the sealing film 220 and the separating film 210 of the functional chamber 150 where the sliding part 310 is located are sequentially broken under the action of the air pressure, and the waste liquid is discharged from the
- the liquid hole 160 flows into the waste liquid tank 600, and the sliding member 300 continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150, and stops moving, so that the pushing member 400 moves toward the first end 111 and cannot Fix the suction piece.
- the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100, so that the adsorbent and the cleaning solution are mixed to clean the adsorbent carrying the target substance.
- the pushing member 400 is moved toward the second end 113 and partially received in the receiving groove 322 to push the sliding member 300 and fix the suction member.
- the sliding member 300 moves toward the second end 113 by the pushing member 400, and the sealing film 220 and the separating film 210 of the functional chamber 150 where the sliding part 310 is located are sequentially broken under the action of the air pressure, and the waste liquid is discharged from the
- the liquid hole 160 flows into the waste liquid tank 600, and the sliding member 300 continues to move, so that the adsorption member carrying the target substance enters the end chamber 140, and stops moving, so that the pushing member 400 moves toward the first end 111 and cannot be fixed Sorption parts.
- the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100, so that the adsorbent carrying the target substance is mixed with the eluent to separate the target substance from the adsorbent to obtain a pure target substance.
- the magnetic field generating mechanism is turned off.
- the sample outlet 190 is communicated with the detection device.
- the pushing member 400 is moved toward the second end 113 and partially received in the receiving groove 322 to push the sliding member 300 and fix the suction member.
- the sliding member 300 moves toward the second end 113 by the pushing member 400.
- the target substance flows into the detection device under the action of the slider 300 for quantitative and qualitative detection.
- the cylinder 100 is divided into an end chamber 140 and N functional chambers 150 by a plurality of separation membranes 210, and the plurality of separation membranes 210 are all sealingly connected to the cylinder 100, N
- the side walls of the functional chamber 150 are provided with drainage holes 160, and each drainage hole 160 is sealed with a blocking film 220, which can be placed in the functional chamber 150 closest to the first end 111 to be configured as an adsorption target
- the material adsorbent can place the reaction liquid or cleaning liquid in other functional chambers 150; since the slider 300 is disposed near the first end 111 and can slide toward the second end 113, when the target substance needs to be separated ,
- the slider 300 can be accommodated in the functional chamber 150 closest to the first end, an adsorption member can be provided on the side of the slider 300 away from the first end 111, and the sample to be separated is added to make the target substance adsorb to the adsorption member On the top, slide the slider 300 toward the second end 113 to rupture
- the above-mentioned separation device 10 is simple, and the separation and detection of the target substance can be achieved without repeatedly opening the cover, the operation is convenient, and the sample to be tested is less polluted.
- the above separation device 10 can be configured for nucleic acid extraction, chemiluminescence detection, or cell sorting.
- the separation film 210 is not limited to three, but may be more, for example, four or five.
- the waste liquid tank 600 is not limited to the above-mentioned pointed shape, and may also be a ring-shaped hollow structure.
- the cylinder 100 is at least partially accommodated in the waste liquid tank 600 and is sealedly connected to the waste liquid tank 600. It can be understood that the waste liquid tank 600 may also be omitted. When the waste liquid tank 600 is omitted, the waste liquid flowing out of the drain hole 160 can directly flow into the external waste liquid collector.
- the detection system is not limited to the PCR (polymerase chain reaction) detection system, but can also be other detection systems, for example: please refer to FIG. 4 together.
- the detection system is a chemiluminescence detection system, including separation Device 10 'and chemiluminescence detection device (not shown).
- the structure of the separation device 10 ' is substantially the same as that of the separation device 10, except that the end chamber 140' is a detection chamber.
- the functional chamber 150 ' closest to the first end 111' is the sample chamber. At least one of the remaining functional chambers 150 'is a reaction chamber.
- the reaction reagent is configured as a reaction product obtained by reacting with the target substance in the sample to be tested, and the reaction product can specifically bind to the detection reagent to emit a signal. Among them, the signal is a light-emitting signal.
- the chemiluminescence detection device is connected to the separation device 10 'to detect the target substance in the sample to be tested.
- the chemiluminescence detection device is a light absorption and analysis device.
- the chemiluminescence detection device is connected to the end chamber 140 'and can receive and analyze the luminescence signal in the end chamber 140' to perform quantitative and qualitative detection of the target substance in the test sample.
- the light-emitting signal may be a fluorescent signal or other signals.
- the sliding part 310' is accommodated in the functional chamber 150 'closest to the first end 111', and the sliding part 310 'is located at the sample port 170' and the air pressure is balanced Between the holes 144 ', add a suction member in the functional chamber 150' closest to the first end 111 ', and position the suction member on the side of the sliding part 310' away from the mounting part 320 ', from the first end 111' Close to the direction of the second end 113 ', the remaining three functional chambers 150' are added with cleaning solution, incubation reagent and cleaning solution in sequence. The detection reagent is added to the end chamber 140 '. It should be noted that, if the sample to be tested needs to be cracked to release the target substance to be adsorbed on the adsorbent, the lysis solution is added to the functional chamber 150 'closest to the first end 111'.
- the sample to be tested is added into the functional chamber 150 'closest to the first end 111' from the sample loading port 170 ', and the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100', so that the sample to be tested is mixed with the adsorbent uniform. If a lysate is added, the sample to be tested is decomposed under the action of the lysate to release the target substance, and the released target substance is adsorbed with the adsorbent to obtain an adsorbent carrying the target substance.
- the pushing member 400 ' is accommodated in the accommodating groove 322' to push the sliding member 300 'and fix the suction member.
- the sliding member 300 ' is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure
- the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150', stops the movement, and causes the pushing member 400 'Move toward the first end 111' without fixing the suction member.
- the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member and the cleaning solution are mixed to clean the adsorption member carrying the target substance.
- the pushing member 400 ' is accommodated in the accommodating groove 322 to push the sliding member 300' and fix the suction member.
- the sliding member 300 ' is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure
- the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150', stops the movement, and causes the pushing member 400 'Move toward the first end 111' without fixing the suction member.
- the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member and the reaction reagent are uniformly mixed to obtain the adsorption member carrying the reaction product.
- the pushing member 400 ' is accommodated in the accommodating groove 322 to push the slider 300' and fix the suction member.
- the sliding member 300 ' is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure
- the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150', stops moving, and causes the pushing member 400 'Move toward the first end 111' without fixing the suction member.
- the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member and the cleaning solution are mixed to clean the adsorption member carrying the reaction product.
- the pushing member 400 ' is moved toward the second end 113' and partially accommodated in the receiving groove 322 'to push the sliding member 300' and fix the suction member.
- the sliding member 300 ' is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure
- the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the reaction product enters the end chamber 140', stops the movement, and causes the pushing member 400 ' Move toward the first end 111 'without fixing the suction member.
- the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member carrying the reaction product and the detection reagent are mixed and reacted.
- the chemiluminescence detection device collects and analyzes the luminescence signal during the reaction between the adsorbent carrying the reaction product and the detection reagent to perform quantitative and qualitative analysis on the target substance in the sample to be tested.
- the separation device 10 'in the above chemiluminescence detection system has a simple structure, and the target substance can be separated without repeatedly opening the lid. The operation is convenient, and the sample to be tested is less polluted. At the same time, the above chemiluminescence detection system is used to detect the The process can be directly detected without removing the target substance from the separation device 10 ', which further reduces the contamination of the target substance and ensures the accuracy of the detection.
- the reaction reagent may be omitted. It can be understood that the separation film 210 'is not limited to four, but may be more, such as five or six, etc., and may be set according to actual conditions.
- sample outlet 190 may be omitted.
- the detection system is not limited to PCR (polymerase chain reaction) detection system and chemiluminescence detection system, but can also be other detection systems, for example: in other embodiments, the detection system is an electrochemical detection system (not shown) , Including separation device and electrochemical detection device.
- the structure of the separation device and the separation device 10 are substantially the same, the difference is that:
- the adsorbent is a chip.
- the adsorption member is accommodated in the functional chamber closest to the first end and fixed on the sliding member.
- the pusher can be made of non-magnetic material.
- the magnetic field generating mechanism is omitted.
- the electrochemical detection device can collect and analyze the electrical signal of the end chamber to perform quantitative and qualitative analysis on the target substance in the sample to be tested.
- the separation device in the above-mentioned electrochemical detection system has a simple structure, and the target substance can be separated and detected without repeatedly opening the cover. The operation is convenient, and the sample to be tested is less polluted to ensure the purity of the target substance.
- the above separation device 10 is not limited to being configured as a detection system or an electrochemical detection system, but can also be configured into other separation systems, for example: a cell sorting system (not shown) of an embodiment, including a separation device and cells Detection device.
- the separation device of the cell sorting system is almost the same as the separation device 10, except for:
- the cleaning solution in the functional chamber is a cleaning solution or a separation solution capable of cleaning the target cells and ensuring the activity of the target cells.
- the end chamber is configured with a buffer solution to suspend the target cells.
- the adsorption member is a biological magnetic bead.
- the magnetic beads can specifically bind to the recognition sites on the surface of the cells to be separated.
- the magnetic beads are MACS MicroBeads magnetic beads or Dynabeads magnetic beads.
- the target cells in the cells to be separated are hybridoma cells or CAR-T cells (chimeric antigen receptor T cell immunotherapy, English full name Chimeric Antigen Receptor T-Cell Immunotherapy).
- the adsorbent may also be a chip that can specifically bind to the target cell.
- the adsorption member is a chip, the adsorption member is accommodated in the functional chamber closest to the first end and fixed on the sliding member.
- the cell detection device is connected to the separation device to perform quantitative or qualitative separation of target cells in the cells to be separated. Further, the cell detection device is connected to the end chamber to perform qualitative or quantitative detection on target cells in the end chamber. Furthermore, the cell detection device can detect and analyze the concentration, purity, and type of target cells.
- the cell detection device is a spectrophotometer, a Raman spectrometer or a chemiluminescence detector. It should be noted that the cell detection device is not limited to the above-mentioned pointing device, as long as the device capable of quantitatively and qualitatively detecting the target cell can be used as the cell detection device, and can be set according to the actual situation.
- the separation and detection of target cells can be achieved without repeated opening of the cover, the operation is convenient, and the sample to be tested is less polluted to ensure the purity of the target cells.
- the separation device and its application, detection system, electrochemical detection system, and cell sorting system provided in the embodiments of the present application are simple in structure through the separation device in the above cell sorting system, and the target sample can be separated without repeatedly opening the cover And detection, easy operation, less contamination of the sample to be tested.
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Abstract
L'invention concerne un dispositif de séparation et son utilisation, un système de détection, un système de détection électrochimique et un système de tri cellulaire. Le dispositif de séparation comprend un corps cylindrique, un ensemble membranaire et un élément coulissant, le corps cylindrique ayant une première extrémité et une seconde extrémité opposées l'une à l'autre; l'ensemble membranaire comprend une pluralité de membranes de séparation et une pluralité de membranes de blocage, la pluralité de membranes de séparation étant logée dans le corps cylindrique et étant reliée de manière étanche au corps cylindrique, la pluralité de membranes de séparation étant agencées à intervalles de la seconde extrémité à la première extrémité pour diviser le corps cylindrique en une chambre d'extrémité et N chambres fonctionnelles, la chambre d'extrémité étant proche de la seconde extrémité, des parois latérales des N chambres fonctionnelles étant pourvues de trous d'évacuation de liquide, chaque trou d'évacuation de liquide étant bloqué par la membrane de blocage; et l'élément coulissant étant disposé à proximité de la première extrémité et pouvant coulisser vers une direction proche de la seconde extrémité. Le dispositif de séparation ci-dessus entraîne moins de contamination pour un échantillon à analyser.
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