WO2015088201A1 - 자성입자 분리장치 및 이를 이용한 핵산 또는 단백질의 분리 및 정제방법 - Google Patents
자성입자 분리장치 및 이를 이용한 핵산 또는 단백질의 분리 및 정제방법 Download PDFInfo
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- WO2015088201A1 WO2015088201A1 PCT/KR2014/011973 KR2014011973W WO2015088201A1 WO 2015088201 A1 WO2015088201 A1 WO 2015088201A1 KR 2014011973 W KR2014011973 W KR 2014011973W WO 2015088201 A1 WO2015088201 A1 WO 2015088201A1
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- Prior art keywords
- tube
- induction magnet
- hole
- induction
- magnetic
- Prior art date
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- 239000006249 magnetic particle Substances 0.000 title claims abstract description 79
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 37
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 33
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 33
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 26
- 108091005461 Nucleic proteins Proteins 0.000 title claims abstract description 15
- 230000006698 induction Effects 0.000 claims abstract description 115
- 238000000926 separation method Methods 0.000 claims abstract description 44
- 238000000746 purification Methods 0.000 claims abstract description 6
- 230000008878 coupling Effects 0.000 claims description 25
- 238000010168 coupling process Methods 0.000 claims description 25
- 238000005859 coupling reaction Methods 0.000 claims description 25
- 108090000623 proteins and genes Proteins 0.000 claims description 24
- 238000005192 partition Methods 0.000 claims description 22
- 239000000243 solution Substances 0.000 claims description 18
- 238000003780 insertion Methods 0.000 claims description 17
- 230000037431 insertion Effects 0.000 claims description 17
- 238000005406 washing Methods 0.000 claims description 11
- 230000002093 peripheral effect Effects 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 238000002955 isolation Methods 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 2
- 239000013612 plasmid Substances 0.000 description 13
- 239000000696 magnetic material Substances 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 5
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- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 2
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- 229910052710 silicon Inorganic materials 0.000 description 2
- 239000010703 silicon Substances 0.000 description 2
- 230000004544 DNA amplification Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
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- 238000010828 elution Methods 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 238000007373 indentation Methods 0.000 description 1
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Images
Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C1/00—Magnetic separation
- B03C1/02—Magnetic separation acting directly on the substance being separated
- B03C1/28—Magnetic plugs and dipsticks
- B03C1/288—Magnetic plugs and dipsticks disposed at the outer circumference of a recipient
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L9/00—Supporting devices; Holding devices
- B01L9/06—Test-tube stands; Test-tube holders
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C1/00—Magnetic separation
- B03C1/005—Pretreatment specially adapted for magnetic separation
- B03C1/01—Pretreatment specially adapted for magnetic separation by addition of magnetic adjuvants
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C1/00—Magnetic separation
- B03C1/02—Magnetic separation acting directly on the substance being separated
- B03C1/025—High gradient magnetic separators
- B03C1/031—Component parts; Auxiliary operations
- B03C1/033—Component parts; Auxiliary operations characterised by the magnetic circuit
- B03C1/0332—Component parts; Auxiliary operations characterised by the magnetic circuit using permanent magnets
-
- 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
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/405—Concentrating samples by adsorption or absorption
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/025—Align devices or objects to ensure defined positions relative to each other
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L2200/0668—Trapping microscopic beads
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/043—Moving fluids with specific forces or mechanical means specific forces magnetic forces
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C2201/00—Details of magnetic or electrostatic separation
- B03C2201/18—Magnetic separation whereby the particles are suspended in a liquid
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C2201/00—Details of magnetic or electrostatic separation
- B03C2201/26—Details of magnetic or electrostatic separation for use in medical or biological applications
Definitions
- the present invention relates to an apparatus for separating magnetic particles and a method for separating and purifying nucleic acids or proteins using the same.
- nucleic acids DNA and RNA
- proteins proteins
- Isolation of nucleic acids (DNA and RNA) or proteins from biological samples is the most important step in biochemical research and diagnostic processes. If the genetic material (nucleic acid) is not isolated from the sample, the next steps, gene detection, gene cloning, gene sequencing, gene amplification, cDNA synthesis, etc. cannot be performed.
- an effective and reproducible separation method is required, and a separation method using magnetic particles has recently been developed.
- the nucleic acid separation method using magnetic particles is a method of inducing binding to a genetic material using magnetic particles and then separating the sample using an external magnetic field. Magnetic particles generally used to separate and purify DNA, RNA, proteins, etc. It is known that the particle size is about 500-2000 nm.
- US Patent Publication 2013/0026104 (MAGNETIC SEPARATION DEVICE AND METHODS) has been disclosed as a device for applying a magnetic field to the genetic material to separate.
- the central support member 119 is provided with a magnet 123 at an approximately center portion between the upper and lower support portions 111 and 201.
- the upper support portion 201 or the lower support portion 111 is provided with an accommodating portion 501, the magnetic material is accommodated together with the magnetic material.
- the receiving portion 501 is inconvenient, such as screwed to be coupled to the separation device 101, the receiving portion 501 to remove the magnetic field provided on the receiving portion 501 side There is a lot of inconvenience, such as having to pull out again from the separation device (101).
- the present invention has been made to solve the problems of the prior art, so that it is very convenient to apply and remove the magnetic field to the receiving portion (hereinafter referred to as the 'tube') very fast and convenient to separate and purify the desired nucleic acid or protein from the sample To do this.
- the present invention is intended to achieve structural stability by allowing the tube containing the sample to be stably fixed to the apparatus according to the present invention.
- the present invention is a magnetic field is formed a plurality of induction magnets (100) for inducing magnetic particles; An induction magnet fixing part (200) having an induction magnet fixing hole (210) for inserting and fixing each of the induction magnets (100); And the induction magnet fixing part 200 is coupled, and an inflow hole 310 is formed in a height direction to correspond to a position where the induction magnet 100 is arranged, and another inflow adjacent to any one inflow hole 310a is formed. And a body 300 having a partition wall 320 for distinguishing the holes 310b.
- the partition wall 320 is the body 300 by pressing the outer peripheral surface of the insertion hole 321 formed in the longitudinal direction of the partition wall 320, and the tube (T) inserted into the inlet hole (310). And a tube fixing portion 322 inserted into the insertion hole 321.
- the body 300 of the present invention penetrates in the longitudinal direction of the partition wall 320, the groove portion 360 is formed into the inner wall 312 for forming the inlet hole 310; And a tube fixing part 322 that is fixed to the body 300 by pressing an outer circumferential surface of the tube T inserted into the inflow hole 310 and inserted into the groove part 360.
- the bottom surface BS of the tube and the bottom surface 311 of the inflow hole 310 are separated by a predetermined distance t.
- the depth D of the inflow hole 310 may be longer than the length L of the tube T inserted into the inflow hole 310.
- the end 211 of the induction magnet fixing hole 210 may be protruded.
- the induction magnet fixing part 200 of the present invention the first coupling magnet for fixing the first coupling magnet 230 for coupling with the body 300 on the edge side of the surface formed with the induction magnet fixing hole 210
- the hole 220 is formed, and the first coupling magnet fixing hole 220 to be fixed to the induction magnet fixing part 200 by the magnetic force of the first coupling magnet 230 to the body 300,
- a second coupling magnet fixing hole 330 may be formed in the corresponding portion to seat the second coupling magnet 340.
- One side of the induction magnet 100 of the present invention is fixed to the induction magnet fixing hole 210 of the induction magnet fixing part 200, the other side is formed to protrude upwards, protruding from the lower side of the body 300
- a cross-sectional area of the inflow hole 310 may be smaller than an upper side of the inflow hole 310 so that a space in which the induction magnet 100 may be located between the partition wall 320 and another partition wall 320a adjacent thereto is provided.
- the washing and separation solution injection to inject a washing solution or separation solution into the tube (T) to separate the magnetic particles and nucleic acid or protein remaining in the tube (T) Step S700 may be further included.
- FIG. 1 is a conceptual diagram of a magnetic particle separation device according to the present invention
- FIG. 2 is a conceptual diagram illustrating one side decomposition of a magnetic particle separating device according to the present invention
- FIG 3 is an exploded conceptual view of another direction of the magnetic particle separation device according to the present invention.
- Figure 4 is a cross-sectional side view of the magnetic particle separation device according to the present invention.
- FIG. 5 is a cross-sectional view of the upper side of the magnetic particle separator according to the present invention
- Figure 6 is a lateral decomposition conceptual view of another magnetic particle separation device according to the present invention.
- Figure 7 is one side cross-sectional view of another magnetic particle separation device according to the present invention.
- Figure 8 is a cross-sectional side view of a third magnetic particle separation device according to the present invention.
- FIG. 9 is a conceptual diagram of a fourth magnetic particle separation device according to the present invention.
- FIG. 10 is an exploded conceptual view of a fourth magnetic particle separation device according to the present invention.
- FIG. 11 is a cross-sectional side view of a fourth magnetic particle separation device according to the present invention.
- FIGS. 6 and 7 correspond to the second embodiment according to the present invention
- FIGS. 9 to 11 correspond to the fourth embodiment according to the present invention.
- the present invention provides an induction magnet fixing part equipped with a body 300 and an induction magnet 100 provided with a plurality of inflow holes 310 to insert a tube T having a substantially cylindrical shape. 200).
- Induction magnet fixing part 200 is formed in a substantially plate shape is coupled to one side of the body (300).
- the induction magnet 100 of the present invention has an intensity that can form a magnetic field as a whole in a tube T inserted into the body 300 or the body 300 as a magnetic object. It is desirable to have.
- the shape of the induction magnet 100 is illustrated as having a circular shape and a little thickness, like a coin, but the cross-sectional shape may be various shapes such as square, hexagon, and octagonal shape. If the shape can be effectively fixed to the induction magnet fixing hole 210 formed in the 200 is sufficient.
- the induction magnet 100 is inserted into and fixed to the induction magnet fixing holes 210 formed in the induction magnet fixing part 200 to be described below.
- Induction magnet 100 is provided to have a plurality of rows (L1, L2, L3, L4) and columns (C1, C2, C3, C4, C5, C6).
- the tube T is arranged to correspond to any one of the rows C1, C2, ... C6.
- the plurality of rows L1, L2, ... L4 is provided to effectively form a magnetic field on one side of the tube T. Rows and columns in which the induction magnet is arranged can be manufactured in various applications, and are flexible according to the size and number of tubes to be inserted.
- the induction magnet fixing part 200 of the present invention is coupled to the coupling magnet fixing hole 220 and inserted therein for coupling with a plurality of induction magnet fixing holes 210 and the body 300 to be described later
- the magnet 230 is provided.
- the inflow hole 310 of the tube T may also be six so as to correspond thereto. This means that the number of columns of the induction magnet fixing hole 210 is variable according to the number of inflow holes 310.
- the number of the inlet hole 310 is flexible depending on the size and number of the tube (T) to be inserted.
- a portion of the body 300 on which the induction magnet fixing part 200 is mounted may be bent inward to achieve an approximately rectangular parallelepiped shape.
- the first plate 240 protrudes more than the second plate 250.
- the body 300 is provided with an inlet hole 310 into which the tube T is inserted.
- a partition wall 320 for partitioning a specific inflow hole 310a and an inflow hole 310b adjacent thereto is formed.
- the partition wall 320 has an insertion hole 321 formed to penetrate the inside thereof in the longitudinal direction thereof.
- the insertion hole 321 is inserted into the tube fixing portion (322a) in which a part of the outer peripheral surface protrudes into the inlet hole (310).
- the guide magnet fixing part 200 is provided with a first coupling magnet fixing hole 220 and a first coupling magnet 230 inserted therein at approximately its corners.
- the body 300 is provided with a second coupling magnet fixing hole 330 and a second coupling magnet 340 coupled thereto at a position corresponding thereto.
- the first coupling magnet 230 of the guide magnet fixing part 200 and the second coupling magnet 340 of the body 300 may be coupled to the body 300 by mutual magnetic force.
- the receiving unit 350 is formed to allow the first plate 240 and the second plate 250 of the above-described induction magnet fixing part 200 to be seated.
- the body 300 is a transparent material, in particular, it is preferable to use a transparent acrylic material, such as a transparent acrylic material so as to visually determine whether the nucleic acid and the magnetic material is separated from the tube (T) inserted and fixed through the inlet hole (310) Do.
- a transparent acrylic material such as a transparent acrylic material so as to visually determine whether the nucleic acid and the magnetic material is separated from the tube (T) inserted and fixed through the inlet hole (310) Do.
- An insertion hole 321 is drilled in each partition wall 320 formed in the body 300.
- the tube fixing part 322a is inserted into the drilled insertion hole 321, and one side surface of the tube fixing part 322a protrudes into the inlet hole 310.
- the tube fixing part 322a may be made of a rubber or silicone material that does not slip well. In particular, it is preferable to use a silicon O-ring with a hollow inside. This is to prevent damage such as the tube T is broken or crushed by applying excessive pressure toward the tube T due to the tube fixing part 322a when the tube T is inserted.
- the tube (T) is tightly fixed to the body 300 by the tube fixing part 322a using a silicon O-ring, so as not to be easily separated from the body 300.
- the nucleic acid and the magnetic material in the tube (T) is completed by the magnetic field of the induction magnet 100 fixed to the induction magnet fixing part 200, it is easy to turn over the body 300 to remove the residual solution.
- the present invention by using the first and / or second coupling magnets 230 and 340 for easy attachment and detachment of the induction magnet fixing part 200 and the body 300, anyone can easily connect to the induction magnet 100. There is an advantage that can be applied or terminated by the magnetic field to the body 300 side.
- the induction magnet fixing part 200 After the induction magnet fixing part 200 is removed from the body 300 by injecting a washing solution, nucleic acid or protein separation solution into the tube T, and reacting, the induction magnet fixing part 200 is coupled to the body 300.
- the magnetic field When the magnetic field is provided, the magnetic particles in the tube T are attracted to the induction magnet fixing part 200 to have an effect of separating from the supernatant.
- the induction magnet fixing part 200 is easily removing or recovering the separated supernatant, there is an advantage that can be easily separated nucleic acid or protein.
- the magnetic particles used may be magnetic particles produced by the method described in Korean Patent No. 10-1053023 or AccuBead TM manufactured by Bioneer, but is not limited thereto.
- magnetic particles used for protein separation and purification can be used.
- the inlet hole 310 has a shape in which a cross-section is approximately 'C' shaped and one side is open.
- a groove 360 is formed in the longitudinal direction of the body starting from one side of the partition wall 320 to the inner wall 312 of the body.
- the tube fixing part 322b is inserted into the straight groove 360 formed to extend in the width direction.
- the material of the tube fixing part 322b and the like are in accordance with the first embodiment.
- the outer peripheral surface side of the tube T may be pressed.
- the diameter of the tube fixing part 322b is preferably longer than the length of the groove portion 360 formed on the inner wall 312 side of the body. That is, a part of the tube fixing part 322b may protrude from one side portion of the inflow hole 310.
- the bottom surface BS of the tube and the bottom surface 311 of the inlet hole 310 have a predetermined distance t. Can be spaced apart.
- the height of the partition wall 320 formed in the body 300 may be formed lower than the height of the entire body (300).
- a locking jaw P may be formed at an upper end side of the partition wall 320 so that the outer circumferential surface of the tube T may be caught. The catching jaw P of the tube T is caught by the upper end side of the partition wall 320. Alternatively, by increasing the outer diameter of the tube T to the upper side, it may naturally be caught on the upper end side of the partition wall 320.
- the end 211 of the induction magnet fixing hole 210 may be protruded. End 211 protrudes to forcibly push the induction magnet 100 to the induction magnet fixing hole 2100.
- the inlet and the induction magnet 100 of the induction magnet fixing hole 210 having different diameters are formed. Due to this, the induction magnet 100 is secured and firmly fixed to the induction magnet fixing hole 210 side.
- the upper end of the tube T is chamfered inward so that the cap C of the tube T can effectively block the inlet of the tube. For ease of mounting.
- the body 300 may have a form in which its lower surface is open.
- the body 300 may have a height such that the tube T does not touch the bottom surface of the table.
- the number of matrices of the induction magnet 100 is different from that shown in FIG. 2, etc., this is dependent on the size and the number of tubes into which the number of the induction magnets 100 is introduced according to the first to third embodiments. It means that it can be arranged variably.
- the tube fixing part 322a may be understood to be disposed in a similar manner to the first embodiment, but may be replaced with the tube fixing part 322b same as the second embodiment.
- Components not described above among the components of FIG. 8 are according to the first and / or second embodiments.
- an induction magnet 100, an induction magnet fixing part 200 to which the induction magnet 100 is fixed, a body 300, and the like are provided. Additionally, the body position fixing part 400 is further provided therebetween so that the body 300 can be fixed to the induction magnet fixing part 200.
- the guide magnet fixing part 200 is mounted on the side portion of the body 300, but in the second embodiment, the body 300 is fixed to the upper surface side of the guide magnet fixing part 200.
- the induction magnet fixing holes 210 are drilled in the induction magnet fixing part 200 so that the induction magnets 100 may be fixed, respectively.
- the induction magnet 100 may be firmly fixed to the induction magnet fixing hole 210 by using a forced indentation or a bond.
- the induction magnet 100 is preferably in a cylindrical shape having a sufficient height.
- the present application is described by applying to the 96-well plate, but is not limited to this may be applied in various ways.
- the number of induction magnets may be variably changed according to the number of rows and columns constituting a multi-well plate.
- an inlet hole 310 (96 inlet holes) of 8 * 12 is provided, and is partitioned by partition walls 320 therebetween.
- 9 * 13 spaces (117 spaces) are formed in the lower side of the inflow hole 310, one by one than the number of rows and columns of the inflow hole 310.
- the induction magnet 100 is inserted and fixed in the induction magnet fixing part 200 by the number (117) having the same row and column so that the induction magnet 100 can be arranged in each space.
- the cross-sectional area of the inlet hole 310 is preferably smaller than the upper side as shown in FIG.
- the body position fixing part 400 is disposed therebetween so that the body 300 can be fixedly mounted to the induction magnet fixing part 200.
- the body position fixing unit 400 may include a pressing unit 410 and a receiving unit 420 in which the pressing unit 410 is accommodated.
- the pressing unit 410 may use an O-ring made of rubber or silicone. In order to prevent the appearance of the body 300 is broken or crushed when forced insertion and not easily fall out.
- a nucleic acid isolation and purification method using the magnetic particle separation device according to the first to third embodiments will be described.
- magnetic particles are mixed with a genetic material and then injected into a tube (S100).
- the tube T is inserted into the at least one inlet hole 310 drilled in the height direction of the body 300 (S200).
- the tube fixing parts 322a and 322b located inside the body 300 press the outer circumferential surface of the tube T, the tube T is fixed to the body 300 side (S200).
- the induction magnet fixing part 200 to which the induction magnet 100 is fixed is fixed to one side of the body 300 (S300).
- the order of steps S100 to S300 described above may be changed according to the user's convenience.
- Magnetic particles coupled to the genetic material by the magnetic field formed by the induction magnet 100 is attracted to the induction magnet 100 (S400).
- the remaining liquid except for the magnetic particles combined with the attracted genetic material is removed from the tube (S500).
- the magnetic field formed in the body is removed by removing the induction magnet fixing part 200 fixed to one side of the body 300 (S600).
- a washing solution into the tube (T) (S700). This effectively isolates and purifies the nucleic acid.
- Gastric washing and separation solution injection step (S700) may be performed repeatedly one or more times.
- a nucleic acid isolation and purification method using the magnetic particle separation device according to the fourth embodiment will be described.
- magnetic particles are mixed with a genetic material and then injected into a multi-well plate (S100).
- the multi-well plate body 300 is fixed to the body position fixing unit 400 so that the induction magnet 100 can be introduced between the unit wells of the multi-well plate body 300.
- Magnetic particles coupled to the genetic material by the magnetic field formed by the induction magnet 100 is attracted to the induction magnet 100 (S400).
- the remaining liquid except for the magnetic particles combined with the attracted genetic material is removed from the multi-well plate body 300 (S500).
- the magnetic field formed in the body is removed by removing the body position fixing part 400 fixed to the multi-well plate body 300 (S600).
- a washing solution is injected into the unit well of the multi-well plate body 300 (S700).
- Gastric washing and separation solution injection step (S700) may be performed repeatedly one or more times.
- plasmid DNA separation method using the magnetic particle separation device according to the present invention will be described.
- a nucleic acid in addition to the following method, a method for protein separation is widely included.
- a cell disruption solution is prepared.
- Cell disruption solutions can be obtained through chemical cell disruption methods, which includes a wide range of methods used by those skilled in the art.
- the supernatant After injecting the magnetic particles into the prepared cell disruption solution, the supernatant is obtained after combining the cellular protein denatured aggregates and the three fragmented particles and the magnetic particles.
- the method of obtaining the supernatant may also use the magnetic particle separation device of the present invention.
- the magnetic field is applied / released using the magnetic particle separation device according to the present invention for separation of the bound plasmid DNA-magnetic particles, whereby the plasmid DNA-magnetic particles are attracted and attached to the induction magnet fixing side in the tube. At this time, impurities other than the plasmid DNA-magnetic particles can be removed.
- the washing solution is injected into the tube containing the plasmid DNA-magnetic particles from which the impurity is removed, and the washing step is performed.
- the magnetic field using the induced magnetism fixing unit 200 is used.
- the target protein can be separated from the plasmid DNA-magnetic particle conjugate by applying / releasing the plasmid DNA-magnetic magnetic field.
- the present invention can be very useful for extracting only purified nucleic acids or proteins as it is very easy to apply and remove magnetic fields on the tube side.
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Abstract
Description
Claims (10)
- 자기장이 형성되어 자성입자를 유도시키기 위한 다수개의 유도자석(100);상기 유도자석(100) 각각을 삽입, 고정하기 위한 유도자석고정홀(210)을 구비하는 유도자석고정부(200); 및상기 유도자석고정부(200)가 결합되며, 상기 유도자석(100)이 배열된 위치와 대응되도록 높이방향으로 유입홀(310)이 형성되되, 어느 하나의 유입홀(310a)과 인접한 또 다른 유입홀(310b)을 구분시키기 위한 구획벽(320)이 구비된 몸체(300);를 포함하는 것을 특징으로 하는 자성입자 분리장치.
- 제 1 항에 있어서,상기 구획벽(320)은상기 구획벽(320)의 길이방향으로 형성된 삽입홀(321), 및 상기 유입홀(310)에 삽입되는 튜브(T)의 외주면을 압박하여 상기 몸체(300)에 고정시키며, 상기 삽입홀(321)에 삽입되는 튜브고정부(322)를 포함하는 것을 특징으로 하는 자성입자 분리장치.
- 제 1 항에 있어서,상기 몸체(300)는 상기 구획벽(320)의 길이방향으로 관통하며, 상기 유입홀(310)을 형성시키기 위한 내측벽(312)의 내부로 형성되는 홈부(360); 및상기 유입홀(310)에 삽입되는 튜브(T)의 외주면을 압박하여 상기 몸체(300)에 고정시키며, 상기 홈부(360)에 삽입되는 튜브고정부(322);를 포함하는 것을 특징으로 하는 자성입자 분리장치.
- 제 1 항 내지 제 3 항 중 어느 한 항에 있어서,상기 유입홀(310)의 내측으로 상기 튜브(T)가 삽입될 때, 상기 튜브의 바닥면(BS)과 상기 유입홀(310)의 바닥면(311)은 소정거리(t) 이격되도록 상기 유입홀(310)의 깊이(D)는 상기 유입홀(310)의 내측으로 삽입되는 상기 튜브(T)의 길이(L)보다 긴 것을 특징으로 하는 자성입자 분리장치.
- 제 1 항 내지 제 3 항 중 어느 한 항에 있어서,상기 유도자석(100) 각각을 강제 삽입 후 고정시키기 위하여, 상기 유도자석고정홀(210)의 단부(211)는 돌출형성된 것을 특징으로 하는 자성입자 분리장치.
- 제 1 항 내지 제 3 항 중 어느 한 항에 있어서,상기 유도자석고정부(200)에는 상기 유도자석고정홀(210)이 형성된 면의 가장자리측에는 상기 몸체(300)와 결합시키기 위한 제1결합자석(230)을 안착시키기 위한 제1결합자석고정홀(220)이 형성되며,상기 몸체(300)에 상기 제1결합자석(230)의 자력에 의하여 상기 유도자석고정부(200)가 고정될 수 있도록, 상기 제1결합자석고정홀(220)과 대응되는 부분에 제2결합자석(340)을 안착시키기 위한 제2결합자석고정홀(330)이 형성되는 것을 특징으로 하는 자성입자 분리장치.
- 제 1 항에 있어서,상기 유도자석(100)의 일측은 상기 유도자석고정부(200)의 유도자석고정홀(210)에 고정되고, 타측은 상방향으로 돌출 형성되며,상기 몸체(300)의 하부측에는 돌출 형성된 상기 유도자석(100)이 상기 구획벽(320)과 인접한 또 다른 구획벽(320a) 사이에 위치할 수 있는 공간이 마련되도록 상기 유입홀(310)의 단면적은 상부측보다 하부측이 작은 것을 특징으로 하는 자성입자 분리장치.
- 제 7 항에 있어서,상기 몸체(300)와 상기 유도자석고정부(200)사이에는,상기 몸체(300)가 상기 유도자석(100)이 고정된 상기 유도자석고정부(200)에 고정될 수 있도록 상기 몸체(300)의 외주면을 압박시키기 위한 압박부(410); 및 상기 압박부(410)를 내측부에 수용시켜 고정하기 위한 수용부(420);를 포함하는 몸체위치고정부(400)를 더 포함하는 것을 특징으로 하는 자성입자 분리장치.
- 제 1 항 또는 제 8 항의 자성입자 분리장치를 이용한 핵산 또는 단백질의 분리 및 정제방법에 있어서,자성 입자를 이용한 핵산의 분리를 위하여 유전자 물질에 자성입자를 섞은 후 튜브(T)에 주입하는 주입단계(S100);상기 몸체(300) 측에 상기 튜브(T)를 고정시키는 튜브고정단계(S200);유도자석(100)이 고정된 유도자석고정부(200)를 몸체(300)의 일측에 고정시키는 유도자석고정부결합단계(S300);상기 유도자석(100)에 의하여 형성된 자기장에 의하여 상기 유전자 물질과 결합된 자성입자가 상기 유도자석(100)측으로 유인되는 자성입자유인단계(S400);상기 유인된 유전자 물질과 결합된 자성입자를 제외한 잔여액체를 상기 튜브(T)로부터 제거하는 잔여액체제거단계(S500); 및상기 몸체(300)의 일측에 고정된 유도자석고정부(200)를 제거하여 몸체에 형성된 자기장을 제거하는 자기장제거단계(S600);을 포함하는 것을 특징으로 하는 핵산 또는 단백질의 분리 및 정제방법.
- 제 9 항에 있어서,상기 자기장제거단계(S600) 이후에는,상기 튜브(T)에 잔류하는 자성입자와 핵산 또는 단백질을 분리하기 위하여 상기 튜브(T) 내측으로 세척용액 또는 분리용액을 주입하는 세척 및 분리용액주입단계(S700)를 더 포함하는 것을 특징으로 하는 핵산 또는 단백질의 분리 및 정제방법.
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JP2016538731A JP6360559B2 (ja) | 2013-12-09 | 2014-12-05 | 磁性粒子分離装置、並びに、この装置を用いる核酸またはたんぱく質の分離及び精製方法 |
CN201480067615.4A CN105916972B (zh) | 2013-12-09 | 2014-12-05 | 磁性粒子分离装置及利用该装置的核酸或蛋白质的分离及提纯方法 |
US15/103,102 US10441957B2 (en) | 2013-12-09 | 2014-12-05 | Magnetic particle separating device, and method of separating and purifying nucleic acid or protein using same |
EP14869081.1A EP3081634B1 (en) | 2013-12-09 | 2014-12-05 | Magnetic particle separating device |
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US (1) | US10441957B2 (ko) |
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JP (1) | JP6360559B2 (ko) |
KR (1) | KR102371946B1 (ko) |
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WO2022114917A1 (ko) | 2020-11-30 | 2022-06-02 | (주)바이오니아 | 중저 비유전율을 가진 화합물을 포함한 핵산 결합 완충액을 이용한 핵산 분리방법 |
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EP4286516A3 (en) * | 2015-02-24 | 2024-03-06 | 10X Genomics, Inc. | Partition processing methods and systems |
CN106754345A (zh) * | 2016-12-02 | 2017-05-31 | 上海默里科基因科技有限公司 | 核酸提取装置 |
RU2697275C1 (ru) * | 2018-03-19 | 2019-08-13 | Общество с ограниченной ответственностью "Синтол" (ООО "Синтол") | Магнитный штатив для пробирок для выделения и очистки нуклеиновых кислот и/или макромолекул из биологического образца |
US11242519B2 (en) * | 2018-08-23 | 2022-02-08 | Alpaqua Engineering, LLC | Discontinuous wall hollow core magnet |
KR102192403B1 (ko) * | 2019-02-13 | 2020-12-17 | 주식회사 이지다이아텍 | 자동 단백질 정제 장치 및 이를 이용한 단백질 정제 방법 |
CN110317712A (zh) * | 2019-07-17 | 2019-10-11 | 英诺维尔智能科技(苏州)有限公司 | 一种自动化磁体细胞分离分选装置 |
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CN105916972B (zh) | 2018-12-21 |
CN105916972A (zh) | 2016-08-31 |
US10441957B2 (en) | 2019-10-15 |
KR20150067048A (ko) | 2015-06-17 |
JP2017508599A (ja) | 2017-03-30 |
KR102371946B1 (ko) | 2022-03-08 |
EP3081634A4 (en) | 2017-09-06 |
US20160368001A1 (en) | 2016-12-22 |
JP6360559B2 (ja) | 2018-07-18 |
EP3081634A1 (en) | 2016-10-19 |
EP3081634B1 (en) | 2023-12-20 |
EP3081634C0 (en) | 2023-12-20 |
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