Device and method for mixing magnetic particles with a fluid

• US 6,764,859 B1
• Filed: 06/04/2002
• Issued: 07/20/2004
• Est. Priority Date: 07/19/1999
• Status: Expired due to Term

• Current Assignee: Biomerieux B.V. (Compagnie Merieux Alliance SAS)
• Original Assignee: Biomerieux B.V. (Compagnie Merieux Alliance SAS)
• Inventors: Verwimp, Emiel Gerebern Maria, Kreuwel, Hermanus Johannes Maria
• Primary Examiner(s): Ludlow, Jan M.
• Application Number:
  US10/031,222
• Time in Patent Office: 777 Days
• Field of Search: 536/25.4, 366/273, 366/274, 209/213-217, 209/225-227, 435/287.1, 435/287.2, 435/287.3, 435/6, 436/526, 436/178, 436/94, 422/99, 422/101, 422/63-67, 210/695, 210/222
• US Class Current: 436/178
• CPC Class Codes: B01F 33/451 wherein the mixture is directly exposed to an electromagnetic field without use of a stirrer, e.g. for material comprising ferromagnetic particles or for molten metal
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/0332 using permanent magnets
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 for use in medical applications
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/143 Magnetism, e.g. magnetic label
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

This invention relates to the use of magnetic or magnetizable particles, and, in particular, to methods of mixing magnetic or (super)paramagnetic particles efficiently with a fluid and the separation of the magnetic particles from a fluid, optionally followed by resuspension of the particles in another fluid. The present invention provides a method of mixing, in one or more container(s), magnetic or (super)paramagnetic particles with a fluid, using more than one magnets, whereby the containers are subjected to magnetic fields with different and changing directions by moving the magnets with respect to the position of the container(s) and/or by moving the containers with respect to the positions of the magnets. The invention further provided a device for doing the same. Preferably the holders for the containers and the magnets in the device are placed in intervening array geometrics and the magnets are placed in line in such a way that all magnets that are in line have their poles oriented in the same direction, and that all magnets in a neighboring line have their poles oriented in the reverse direction with respect to the poles of the magnets in the first line.

Claims

1. Method of mixing, in one or more container(s), magnetic or (super) paramagnetic particles with a fluid, using at least two magnet arrays with a corresponding container array disposed between adjacent magnet arrays to form an intervening array geometry, wherein each magnet array comprises a plurality of spaced apart magnets, whereby the containers are subjected to magnetic fields with different and changing directions by moving the magnets in the adjacent magnet arrays with respect to the position of the container(s) in the corresponding container array and/or by oscillating the containers with respect to the positions of the magnets.

2. Method according to claim 1, wherein the containers, by oscillating the containers or moving the magnets in first and second adjacent magnet arrays, are subjected to magnetic fields of opposite polarity.

3. Method according to claim 1, wherein, as a result of moving either the magnets or the containers, the magnetic or (super) paramagnetic particles in the fluid in respective containers are repeatedly moved between two corresponding magnets that face each other with the same pole, the corresponding magnets being disposed in adjacent magnet arrays on opposing sides of the containers with one held in a first magnet array and the other held in a second magnet array.

4. Method according to claim 1, wherein the magnets in a respective magnet array are moved in concert with respect to the position of the containers and/or the containers in a respective container array are moved in concert with each other with respect to the position of the magnets in such a way that the magnetic or (super)paramagnetic particles are moved through the fluid to one side of the container by bringing a first magnet with its magnetic pole close to the wall of the container and, subsequently are moved to the opposite side by bringing a second magnet close to the opposite wall of the container, whereby said second magnet has the same magnetic pole as the first magnet in such a way that the direction of the magnetic field in each container is repeated reversed.

5. Method according to claim 1, wherein the magnets of respective magnet arrays are moved in concert with respect to the containers.

6. Method according to claim 1, further comprising dispersing the magnetic or (super)paramagnetic particles in a clotted or aggregate configuration into the fluid.

7. Method according to claim 6, further comprising mixing the fluid sufficiently to cause the clotted and/or aggregate of magnetic particles to separate and/or disperse to thereby promote mixing with the fluid.

8. Method according to claim 1, further comprising linearly translating magnets in respective magnet arrays about substantially parallel paths so that the magnets in each array move forward and rearward in concert and so that adjacent arrays of magnets move in opposing directions on opposite sides of a respective container array in such a way that a direction of the magnetic field in each container is repeatedly substantially reversed.

9. Method according to claim 1, wherein the containers are oscillated by moving the containers in the container array in concert back and forth magnet arrays.

10. Method according to claim 1, wherein the at least two magnet arrays comprise first and second magnet arrays with the first container array disposed substantially centrally spaced therebetween, with the first and second magnet arrays and the container array being arranged in substantially parallel rows, with the first magnet array and the second magnet array configured to move in opposite directions alongside the row of containers to provide the mixing.

11. Method according to claim 1, wherein the at least two magnet arrays comprise first and second arrays with the first container array disposed substantially centrally spaced therebetween, and wherein the mixing is carried out by oscillating the containers in the container array, the oscillating step comprises moving the containers in the container array so that containers in the first container array are repeatedly exposed to different magnetic fields provided by the first and second magnet arrays.

12. Device for mixing magnetic or (super) paramagnetic particles in containers with a fluid, said device comprising means for holding said one or more containers in at least one container array having a plurality of spaced apart containers and a plurality of magnet arrays, a respective one disposed on opposing sides of the at least one container array, each magnet array comprising a plurality of spaced apart magnets, the magnet arrays configured and aligned in the device so as to cooperate with the at least one container array to concurrently expose the containers therein to different and changing magnetic field directions and means for horizontally moving said magnets with respect to the position of said containers and/or means for horizontally oscillating said containers with respect to the position of said magnets in such a way that the containers are subjected to magnetic fields with different and changing directions.

13. Device according to claim 12, the device being provided with a heat block that is positioned in such a way that it can be moved into close proximity with the containers so as to warm their contents, and moved away again.

14. Device according to claim 13, wherein the heat block is positioned underneath the containers and has wells which enclose the tips of the containers when the heatblock is brought into close proximity with the container.

15. Device according to claim 12 wherein each magnet in a respective magnet array is oriented in such a way that it repels each of its neighboring magnets.

16. Device according to claim 12, wherein magnets in first and second magnet arrays can be moved back and forth in concert on straight parallel paths along opposite sites of a corresponding first container array in such a way that the direction of the magnetic field in each container in the first container array is repeatedly reversed.

17. Device according to claim 12, wherein the magnets in first and second magnet arrays are placed in line in such a way that all magnets that are in line in the first array have their poles oriented in the same direction, and that all magnets in a neighboring second magnet array line have their poles oriented in the reverse direction with respect to the poles of the magnets in the first magnets array line.

18. Device according to claim 12, herein the magnets can also be moved in a vertical direction so as to positioned at different heights with respect to the walls of the containers.

19. Device according to claim 12, wherein the containers are part of a closed system whereby the containers are adapted to remain in position in the device and serially receive and expel fluid samples.

20. Device according to claim 12, wherein the containers are tube-shaped vessels provided with a tip with a smaller diameter.

21. Device according to claim 12, wherein, in operation, the device is configured to isolate nucleic acid.

22. Device according to claim 12, wherein the device is configured to disperse magnetic or (super) paramagnetic particles in a clotted or aggregate configuration in the fluid.

23. Device according to claim 22, wherein, in operation, the device is configured to cause the clotted and/or aggregate of magnetic particles to separate to thereby promote mixing with the fluid.

24. Device according to claim 12, wherein said device comprises means for moving said magnets, said means for moving the magnet being configured to linearly translate magnets in respective magnet arrays about substantially straight parallel paths so that the magnets in each array move forward and rearward in concert and so that adjacent arrays of magnets move on opposite sides of a respective container array so that the magnetic field direction in each container is repeatedly reversed.

25. Device according to claim 12, wherein said device comprises means for oscillating the containers by moving the containers in the at least one container array in concert back and forth.

26. Device according to claim 12, wherein the plurality of magnet arrays comprise first and second magnet arrays with the at least one container array comprising a first container array that is disposed substantially centrally spaced therebetween, and wherein the magnets are moved to provide the mixing by moving the magnets in the first magnet array forward while rearward moving the magnets in the second magnet array so that adjacent containers in the first container array are alternately exposed to magnetic fields of different pole directions as provided by the magnets in the first and second arrays.

27. Device according to claim 12, wherein the plurality of magnet arrays comprise first and second arrays and the at least one container array comprises a first container array with the first container array disposed substantially centrally spaced therebetween, and wherein the mixing is carried out by oscillating the containers in the first container array, the oscillating step comprises moving the containers in the first container array that containers in the first container array are alternately exposed to changing magnetic fields provided by magnets in the first and second magnet arrays.

28. Method for the isolation of nucleic acid from starting material comprising the following steps:

(a) bringing starting material together with an appropriate lysis buffer and magnetisable particles into at least one container held as one of a plurality of containers in a row that defines a container array that holds the containers in spaced apart alignment,

(b) mixing the content of the at least one container by moving first and second magnet arrays, each magnet array comprising a plurality of spaced apart magnets, with the containers in the container array held between the first and second magnet arrays in such a way that the direction of the magnetic field associated with the at least one container is repeatedly reversed for a sufficient amount of time with the magnets at a height that is adjusted to the volume of the sample,

(c) collecting the particles at a wall of the container using the magnets,

(d) removing most of the sample liquid from the device,

(e) adding a sufficient amount of washing buffer to the device,

(f) repeating step (b) to (d),

(g) adding a suitable amount of elution buffer to the device,

(h) drawing the particles down into the tip of the container by moving the magnets to a lower position,

(i) optionally heating the container by moving a heatblock into close proximity with the containers,

(j) optionally removing an appropriate amount of elution buffer from the device,

(k) repeat step (b),

(l) move the magnets in a vertical direction to a position above the fluid level, and

(m) collect the elution buffer with the isolated nucleic acid container therein.

29. A method of mixing magnetic and/or (super)paramagnetic particles with a fluid comprising:

providing a device with a plurality of containers held in alignment in spaced apart substantially parallel lines of first and second container arrays and a plurality of permanent magnets held in spaced apart substantially parallel lines of first, second and third magnet arrays, with the first container array being positioned between the first and second magnet arrays and the second container array being positioned between the second and third magnet arrays;

introducing magnetic and/or supermagnetic particles into at least one of the containers in the first and/or second container arrays;

moving the magnets in the first magnet array in concert;

concurrently moving the magnets in the second magnet array in concert; and

concurrently moving the magnets in the third magnet array in concert whereby the particles in the containers are exposed to varying magnetic fields and directions and attracted to opposing sides of a respective container to thereby mix the fluid therein.

30. A method of mixing magnetic and/or (super) paramagnetic particles with a fluid in a plurality of containers, comprising:

arranging a plurality of magnet arrays and at least one container array having opposing first and second sides so that the arrays are alignably positioned with a first magnet array disposed on a first side of a first container array and a second magnet array is positioned on a second side of the first container array, wherein the first and second magnet arrays comprise a plurality of discrete spaced apart magnets and the first container array is configured to hold a plurality of containers in spaced apart alignment; and

moving the magnets in the first and second magnet arrays with respect to the position of the container(s) with the magnets in the first and second magnet arrays configured to travel along substantially parallel travel paths to expose the magnetic particles and/or (super) paramagnetic particles in the containers of the first container array to magnetic fields with different and changing directions.

31. A method according to claim 30, wherein the plurality of magnet arrays comprises first, second, and third magnet arrays, and wherein the at least one container array comprises first and second container arrays with the first container array disposed proximate to and intermediate said first and second magnet arrays and the second container arrays disposed proximate to and intermediate the second and third magnet arrays, whereby the magnets in the first, second and third magnet arrays are repeatedly moved forward and rearward, with the magnet arrays traveling to concurrently mix the particles in the containers.

32. A method according to claim 30, wherein the moving step comprises:

moving the magnets in the first magnet array in concert; and

substantially concurrently moving the magnets in the second magnet array in concert whereby the particles in the containers are exposed to varying magnetic fields and directions and attracted to opposing sides of a respective container to thereby mix the fluid therein.

33. A method according to claim 30, wherein the moving step comprises linearly translating magnets in respective magnet arrays about substantially parallel straight line paths so that the magnets in each array move forward and rearward in concert and so that adjacent arrays of magnets move in opposing directions on opposite sides of a respective container array in such a way that a direction of the magnetic field in each container is repeatedly substantially reversed.

34. A method of mixing magnetic and/or (super) paramagnetic particles with a fluid in a plurality of containers, comprising:

arranging a plurality of magnet arrays comprising first, second and third magnet arrays and a plurality of container arrays comprising first and second container arrays, with the container arrays having opposing first and second sides so that a first magnet array is disposed on a first side of the first container array and the second magnet array is positioned on the second side of the first container array and the first side of the second container array and the third magnet array is positioned on the second side of the second container array, wherein the first, second, and third magnet arrays comprise a plurality of discrete linearly aligned spaced apart magnets and the first and second container arrays are configured to hold a plurality of containers in spaced apart linear alignment; and

moving the containers in the first container array with respect to the positions of the magnets in the first and second magnet arrays and the containers in the second container array with respect to the positions of the magnets in the second and third magnet arrays to expose the magnetic particles and/or (super) paramagnetic particles in the containers thereof to magnetic fields with different and changing directions to thereby mix the fluid therein.

35. A method according to claim 34, wherein the step of moving the containers comprises retracting the containers along a predetermined travel path.

36. A method according to claim 34, wherein the step of moving the containers comprises oscillating the containers between selected positions.

37. A device for mixing magnetic and/or (super) paramagnetic particles with a fluid in a plurality of containers, comprising:

a plurality of laterally spaced apart magnet arrays including at least first, second and third magnet arrays and a plurality of container arrays including at least first and second container arrays, each container array having opposing first and second sides with the first magnet array being proximately positioned on the first side of the first container array and the second magnet array being proximately positioned on the second side of the first container array and the first side of the second container array and the third magnet array being positioned on the second side of the second container array, wherein the first, second, and third magnet arrays comprise a plurality of discrete spaced apart magnets and the first and second container arrays are configured to hold a plurality of containers in spaced apart alignment; and

at least one mechanism that moves the magnets in the first, second and third magnet arrays with respect to the position of the container(s) in the first and second container arrays to concurrently expose the magnetic particles and/or (super) paramagnetic particles in the containers to magnetic fields with different and changing directions.

38. A device for mixing magnetic and/or (super) paramagnetic particles with a fluid in a plurality of containers, comprising:

a plurality of laterally spaced apart magnet arrays including at least first and second magnet arrays and a plurality of container arrays including at least first and second container arrays, each container array having opposing first and second sides with the first magnet array being proximately positioned on the first side of the first container array and the second magnet array being proximately positioned on the second side of the first container array and the first side of the second container array, wherein the magnet arrays comprise a plurality of discrete spaced apart magnets and the first and second container arrays are configured to hold a plurality of containers in spaced apart alignment; and

at least one mechanism that moves the containers in the first and second container arrays relative to the magnets in the first and second magnet arrays to concurrently expose the magnetic particles and/or (super) paramagnetic particles in containers in the first and second container arrays to magnetic fields.

39. A device according to claim 38, further comprising a third magnet array with the third magnet array being positioned on the second side of the second container array.

40. A device according to claim 38, wherein the at least one mechanism is configured to advance the containers along a predetermined travel path.

41. A device according to claim 38, wherein the at least one mechanism is configured to retract the containers in concert along a predetermined travel path.

42. A device according to claim 38, wherein the at least one mechanism is configured to oscillate the containers between predetermined positions.