EP1644120B1 - Vorrichtung und verfahren zum abtrennen von magnetischen oder magnetisierbaren partikeln aus einer flüssigkeit - Google Patents

Vorrichtung und verfahren zum abtrennen von magnetischen oder magnetisierbaren partikeln aus einer flüssigkeit Download PDF

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Publication number
EP1644120B1
EP1644120B1 EP04740646A EP04740646A EP1644120B1 EP 1644120 B1 EP1644120 B1 EP 1644120B1 EP 04740646 A EP04740646 A EP 04740646A EP 04740646 A EP04740646 A EP 04740646A EP 1644120 B1 EP1644120 B1 EP 1644120B1
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EP
European Patent Office
Prior art keywords
magnetic field
magnetic
bar
liquid
permanent magnet
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
EP04740646A
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German (de)
English (en)
French (fr)
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EP1644120A1 (de
Inventor
Lothar A Brassard
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
PerkinElmer Chemagen Technologie GmbH
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Chemagen Biopolymer Technologie AG
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Publication date
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Publication of EP1644120A1 publication Critical patent/EP1644120A1/de
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Classifications

    • 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
    • B03C1/286—Magnetic plugs and dipsticks disposed at the inner circumference of a recipient, e.g. magnetic drain bolt
    • 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
    • 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
    • B03C1/04—Magnetic separation acting directly on the substance being separated with the material carriers in the form of trays or with tables
    • B03C1/06—Magnetic separation acting directly on the substance being separated with the material carriers in the form of trays or with tables with magnets moving during operation
    • 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 invention relates to devices for separating magnetic or magnetizable particles from liquids by means of a magnetic field generated by one or more permanent magnets.
  • the invention further relates to methods for separating magnetic or magnetizable particles from liquids by means of a magnetic field generated by one or more permanent magnets.
  • the devices and methods are suitable for, for example, applications in biochemistry, molecular genetics, microbiology, medical diagnostics and forensic medicine.
  • the basic principle of the magnetic separation of substances from complex mixtures is based on the fact that magnetic particles (magnetizable or magnetically attractable particles) are specifically functionalized for the intended separation process, ie they are equipped by chemical treatment with specific binding properties for the target substances to be separated.
  • the size of such magnetic particles is generally in the range of about 0.05 to 500 microns.
  • Magnetic particles which have specific binding properties for certain substances and are suitable for separating these substances from complex mixtures are described, for example, in US Pat DE 195 28 029 A1 and are commercially available (eg, Chemagen Biopolymer-Technologie AG, DE-52499 Baesweiler).
  • the functionalized magnetic particles are added in a first step ("binding step") to a mixture to be purified, which contains the target substance (s) in a liquid which favors the binding of the target substance molecules to the magnetic particles (binding buffer).
  • binding step a first step
  • these magnetic particles are immobilized by application of magnetic forces or a magnetic field, for example by means of a permanent magnet, at a location of the reaction vessel inner wall ("pellet").
  • pellet the liquid supernatant is separated and discarded, for example by suction or decantation. Since the magnetic particles are immobilized in the aforementioned manner, it is largely prevented that these particles are separated with the supernatant.
  • the immobilized magnetic particles are resuspended again.
  • an elution liquid or an elution buffer is used, which is suitable to dissolve the bond between the target substance (s) and the magnetic particles, so that the target substance molecules of the magnetic particles are released and can be separated with the elution liquid, while the magnetic particles are immobilized by the action of a magnetic field.
  • one or more washing steps can be carried out.
  • DE 296 14 623 U1 discloses a magnetic separator equipped with movable permanent magnets.
  • the magnetic particles containing reaction vessel by mechanical drive means to move relative to a permanently installed permanent magnet.
  • DE 100 63 984 A1 described device having a movable magnetic holder and a movable reaction vessel holder.
  • the magnetic particles are immobilized or collected on the inner wall or at the bottom of a reaction vessel as a "pellet".
  • these devices are not suitable for removing the magnetic particles from a reaction vessel. Therefore, to separate the liquid from the magnetic particles in each case the liquid must be sucked from the individual reaction vessels. This is disadvantageous because it involves a high material consumption (disposable pipette tips). Furthermore, it can not be prevented that individual magnetic particles are sucked with; This results in a high error rate. Further errors can be caused by dripping liquids, which lead to cross contamination.
  • a magnetic separator having a plurality of rotatable rods magnetized by an electromagnetic excitation coil can.
  • the magnetic particles By immersing the rod in the magnetic particles containing liquid and pulling out the rod in the magnetized state, the magnetic particles can be removed from the liquid and optionally transferred to another reaction vessel. There they can be released by switching off the excitation coil back into a liquid, for example in a washing or elution liquid.
  • a disadvantage of this device is that the magnetic field generated by the excitation coil is not sufficiently homogeneous, so that the individual rods - depending on their position within the annular exciter coil - are magnetized differently strong. This disadvantage occurs especially when a large number of rods is needed.
  • the exciter coil has a relatively large amount of space, resulting in constructive restrictions.
  • the known devices are not suitable for the simultaneous treatment of larger numbers of samples, as required for high-throughput applications (eg 364 or 1536 well plates).
  • the invention therefore an object of the invention to provide devices and methods with which the separation of magnetic particles from liquids and the transfer of magnetic particles from a liquid is made possible in another liquid, without causing the aforementioned disadvantages occur.
  • the devices and methods should be suitable for use in high throughput processes.
  • the two legs are made of a soft magnetic magnetic material, for example of soft iron (especially Fe-Ni alloys) or magnetizable steel.
  • Their cross section may be, for example, square, rectangular, circular or oval; The size of the cross-sectional area depends on the desired cross-sectional area of the magnetic field and may be 20 to 100 cm 2 , for example. It is also possible, the legs on a frame or attach housing made of non-magnetizable material.
  • the two legs are generally stacked with the leg bearing the head over that portion of the other leg intended to receive the liquid containers (i.e., the sample cups).
  • the header may be detachably arranged, thereby enabling, for example, the replacement of headers having different numbers or types (length, diameter, fixed or movable) of magnetizable bars.
  • the number of rods depends on the number of samples or fluid containers to be treated simultaneously.
  • Microtiter plates, in particular with 96, 384 or 1536 depressions ("wells") are preferably used as containers, so that corresponding head pieces with, for example, 96, 384 or 1536 magnetizable bars are provided for these cases.
  • sample tubes or reaction vessels having a volume of, for example, 0.015 to 100 ml, which can be treated individually or in groups, in each case in combination with magnetizable bars adapted thereto.
  • the rods are also made of a soft magnetic material, as described above. Their length and diameter depend on the intended application, in particular on the dimensions of the containers and the volumes of liquid, and can be varied accordingly. It is further provided that on the rods in each case a strippable, replaceable sheath is plugged in order to avoid cross-contamination between different liquid samples. This is preferably a special Device provided which allows the automatic discarding of used cases and the provision and installation of new cases.
  • a substantially homogeneous magnetic field is generated between the poles of the legs. This allows the arrangement of a larger number of rods, for example in several rows, with the magnetic field at each of the rods being approximately equal; This is particularly advantageous with regard to high throughput methods.
  • the devices according to the invention have the further advantage that the magnetic particles - when switched on - ansanneln substantially at the top of the rods.
  • the permanent magnets are / are movably arranged relative to the magnetic circuit of the device, so that the magnetic field between the poles can be alternately switched on and off by movement of the magnet (s).
  • the magnet (s) is / are in the magnetic circuit or in this in or out of the magnetic circuit out. That is, the magnetic field between the poles is turned on when the permanent magnet (s) are in a first position, and that the magnetic field between the poles is turned off when the permanent magnet (s) in a second position is / are located. In the said second position, the magnet (s) is / are preferably located outside the magnetic circuit.
  • the magnetic field may preferably be switched on and off in such a way that the magnet (s) are moved within the iron circle (magnetic circuit) (eg by rotation) or that the magnet (s) of outside is moved into the magnetic circuit ("turn on") and then moved back outward (“turn off”).
  • the device can be used to remove magnetic particles using the magnetizable rods of a first liquid and to transfer in a second or more liquid and release there again. Furthermore, this makes it possible to additionally use the rods for other functions, for example as stirring rods.
  • all hard magnetic materials known to the person skilled in the art can be used for producing the permanent magnets, in particular ferrites, Al-Ni-Co alloys and rare earth magnets (preferably NdFeB); Such magnetic materials and magnets are commercially available from various manufacturers.
  • the region of the device in which the movable magnet (s) is / are arranged in the iron circle is at least partially surrounded by a material shielding the magnetic field.
  • a material shielding the magnetic field As a shielding material, a soft magnetic material may be used and / or a person skilled in the art, magnetic fields shielding material such. Tinplate or mumetal.
  • This shielding material is arranged around the movable magnet (s) in such a way that when it is turned off no magnetic forces can act on the sample liquid reservoir located in the air gap of the magnetic circuit.
  • a shield which completely surrounds the region in which the permanent magnet (s) are arranged. In particular, this may be formed a short-circuit ring.
  • the device is preferably arranged so that that portion of the device in which the movable magnet (s) are / are arranged in the iron circle, in the case of a movement of the magnet (s) in the iron or at least partially of the magnetic Field shielding material is surrounded.
  • the two legs of the device are connected to each other at the two poles side by a likewise magnetizable (soft magnetic) material, so that a magnetic circuit or a magnetizing ring is formed, which - with the exception of the air gap between the poles - is completely closed.
  • the movable permanent magnet (s) is / are preferably disposed between the two legs and at the other end (i.e., opposite the poles). If the two legs are interconnected as described, then the permanent magnet (s) is / are preferably mounted in or on the region connecting the two legs. Preferably, the magnet (s) are movably installed in a recess provided in one of the legs or in the part connecting the two legs.
  • the magnet In order to enable movement of the permanent magnet (s) for switching the magnetic field on and off, the magnet, or a group of several magnets, can be arranged to be rotatable or tiltable in a recess provided therefor. By turning or tilting the magnet can be brought into a position in which its poles or its magnetic field in the direction of the magnetic circuit or in the direction of the legs have (switched state, maximum field strength between the poles of the legs), or the magnet can be moved to another position, in the magnetic field emanating from it is substantially perpendicular to the said direction (switched-off state).
  • the magnet (s) may also be rotated or tilted to intermediate positions to cause the field strength between the poles of the legs to be less than the maximum value.
  • the movement can be done either directly or indirectly by hand or by means of one or more electric motors, or by pneumatic or hydraulic means; also combinations of the mentioned means are possible.
  • the drive means may further, known in the art means such.
  • B. linkage or gear include.
  • the extent of movement of the / the permanent magnet is predictable.
  • the magnetic field strength depending on the particular application, set to a specific value. This can be achieved in particular by predetermining and adhering to a specific tilting or rotation angle or a specific displacement distance.
  • the head carrying the magnetizable rods is movably mounted.
  • the head piece may be movable in the horizontal plane.
  • drive means eg., Electric, pneumatic, hydraulic
  • the said magnetizable rod (s) are rotatably mounted (about their longitudinal axis) on the respective header and can be rotated during the treatment of a liquid containing magnetic particles to effect mixing to accelerate the detachment of the particles from the rods.
  • the rotation is preferably carried out by electromotive means.
  • liquids containing such particles are introduced into the air gap of the device, below the magnetizable rods, wherein containers of the type mentioned above can be used.
  • at least one holding device is provided which can be positioned below the bars, so that the bars are aligned with the openings of the container.
  • This holding device can be configured for example in the form of a holding plate.
  • the holding device is movable in a substantially horizontal plane in one or more directions; Alternatively or additionally, the holding device may be movable in the vertical direction.
  • the movement is preferably carried out by electromotive drive or by pneumatic or hydraulic means, or by combinations of these means.
  • the holding devices can also be designed in such a way that they are suitable for carrying out shaking movements.
  • the design measures required for this purpose are generally known to the person skilled in the art. It is further provided that both the head piece and the holding device can be moved and used to carry out shaking movements. In this way, a particularly effective mixing of the sample liquid is possible if the rods are immersed therein.
  • a control or regulating device is provided, by means of which the vertical movement of the holding device (s) is adjustable or controllable in such a way that, upon an upward movement, the rods (7) are immersed in the liquid-filled containers (10). is effected.
  • the said holding device may be part of a program-controlled laboratory robot system, wherein it is preferably arranged so that a plurality of individual said containers or groups of such containers, in particular microtiter plates, alternately placed in a position below said rods and then after a predetermined time interval is again brought into a position which is outside the area located below the bars. In this way, a high sample throughput is possible.
  • the devices of the invention may be advantageously combined with other devices for the automated treatment of sample material. Furthermore, two or more of the devices according to the invention can be arranged side by side and combined.
  • the particles into the liquid after step d may be advantageous to release the particles into the liquid after step d by switching off the magnetic field, to mix them, and then by switching them on of the magnetic field to collect again at the bars.
  • the mixing can be effected for example by rotation of the rods or by shaking the holding device and / or the head part.
  • the said methods can be carried out in a particularly simple and rapid manner.
  • the devices and methods of the invention are particularly suitable for the application areas mentioned above, in particular for high-throughput methods.
  • Fig. 1A and 1B show an embodiment of a device according to the invention in a side view.
  • the device (1) has two magnetizable legs (2, 3) of a magnetic circuit, the legs being connected to one another in the region (6). At the opposite end of the legs are the two poles (4,5) with an intermediate air gap (12).
  • the pole (4) of the upper leg (2) carries a head piece (8) with magnetizable bars (7) attached thereto.
  • a holding device (11) which is connected to the pole (5) of the other leg (3) or at least in contact therewith.
  • a sample container (9) is arranged with a plurality of recesses (10) for receiving liquid samples, for example removably fixed on the holding device (11).
  • a rod or cuboid permanent magnet (15) is rotatably arranged.
  • a short-circuit ring (20) is arranged (this is shown broken through in the region of the rotatable magnet).
  • Fig. 1A shows the device in the off state; the position of the permanent magnet (15) is substantially perpendicular to the direction of the magnetic circuit; The magnetic field of the permanent magnet is passed into the short-circuit ring (20).
  • Fig. 1B shows the same device in the on state.
  • the position of the permanent magnet (15) points substantially in the direction of the magnetic circuit.
  • between the poles (4,5) and thus also at the ends of Rods (7) generates a magnetic field that can be used to attract magnetic particles.
  • Fig. 1C shows a sectional view of the in the Fig. 1A / B device, in which by the dashed line a ( Fig. 1B ) indicated level.
  • the arrows (17) show schematically the direction of the magnetic field in the on state.
  • Fig. 1D and 1E show, also in a schematic side view, a further embodiment of the inventive devices, wherein the magnet used has a flat cuboid shape and the poles are located on the two large side surfaces.
  • Fig. 1D is the switched-on state (the magnetic field runs in the direction of the iron circle)
  • Fig. 1E the switched-off state is shown.
  • the position of the short-circuit ring (20) is merely indicated.
  • the others, in Fig. 1A, 1B The elements shown have been omitted for simplicity of illustration.
  • Fig. 2 and Fig. 3 show, also in side view, further construction variants of the devices according to the invention.
  • Fig. 4 shows the device (1) of Fig. 1A / B in plan view; thereby, the annular shape of the short-circuit ring (20) can be seen.
  • the short-circuit ring (20) is designed so that it does not fully abut the magnetic circuit, but a cavity (22) is present. This facilitates access to the rotatable magnet (15).
  • the shorting ring (20) may be composed of two halves (20a, 20b) or multiple parts as indicated by the dashed line 21 to facilitate assembly and disassembly.
  • Fig. 5 shows an embodiment of the device according to the invention (also in side view), in which in the recess (16) a displaceable (double arrow) permanent magnet (15) is provided.
  • Fig. 5 shows the on state in which the permanent magnet causes between the poles (4,5) creates a magnetic field. To turn off the magnet is moved out of the magnetic circuit of the device (1) to the outside.
  • Fig. 6 shows a modification of the in Fig. 1A / B device shown, wherein the two legs (3,4) are of different lengths.
  • Figs. 7A to 7D show various views of a particularly preferred embodiment in which a magnet (15) on a pad (40) is arranged, which is rotatable in a horizontal plane about the axis Y.
  • the magnet (15) can be brought into the magnetic circuit (iron circle) by rotation of the support (40) (switched-on state, Figs. 7C, 7D ) or moved out of the area of the magnetic circuit ( Fig. 7A, 7B ).
  • the pad (40) is preferably in the form of a turntable or as a rotatable arm which is moved by means of known drive devices.
  • two or more magnets may be mounted on the base.
  • Figs. 7A, 7C show a sectional view in the surface of the axis of rotation Y; the Fig. 7B, 7D each show the same device in plan view.
  • Fig. 8 shows an embodiment of the device (1) according to the invention in side view; In this case, the two legs (2,3) are not connected to each other by a common area (6).
  • the rotatable magnet (15) is arranged between the two legs (2, 3), on the side opposite the air gap.
  • the short-circuit ring (20) is shown in sectional view.
  • Fig. 9 shows the front view of the upper leg (4) of a device according to the invention with the head piece (8) and attached rods (7).
  • a holding device (8) is positioned, on which several containers (10) are arranged in rows.
  • the holding device can be moved in the horizontal plane in different directions as well as up and down (arrows).
  • Fig. 10 shows in longitudinal section examples of different forms of magnetizable rods (7).
  • the particles attracted under the influence of the magnetic field are designated by (30).
  • Fig. 9d shows a rod provided with a replaceable sleeve (25).

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  • Physical Or Chemical Processes And Apparatus (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Soft Magnetic Materials (AREA)
  • Magnetic Treatment Devices (AREA)
EP04740646A 2003-07-10 2004-07-05 Vorrichtung und verfahren zum abtrennen von magnetischen oder magnetisierbaren partikeln aus einer flüssigkeit Expired - Lifetime EP1644120B1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10331254A DE10331254B4 (de) 2003-07-10 2003-07-10 Vorrichtung und Verfahren zum Abtrennen von magnetischen oder magnetisierbaren Partikeln aus einer Flüssigkeit
PCT/EP2004/007308 WO2005005049A1 (de) 2003-07-10 2004-07-05 Vorrichtung und verfahren zum abtrennen von magnetischen oder magnetisierbaren partikeln aus einer flüssigkeit

Publications (2)

Publication Number Publication Date
EP1644120A1 EP1644120A1 (de) 2006-04-12
EP1644120B1 true EP1644120B1 (de) 2011-11-30

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EP04740646A Expired - Lifetime EP1644120B1 (de) 2003-07-10 2004-07-05 Vorrichtung und verfahren zum abtrennen von magnetischen oder magnetisierbaren partikeln aus einer flüssigkeit

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Country Link
US (1) US7776221B2 (ja)
EP (1) EP1644120B1 (ja)
JP (1) JP4762139B2 (ja)
AT (1) ATE535305T1 (ja)
CA (1) CA2531399C (ja)
DE (1) DE10331254B4 (ja)
DK (1) DK1644120T3 (ja)
ES (1) ES2377733T3 (ja)
WO (1) WO2005005049A1 (ja)

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CA2531399A1 (en) 2005-01-20
EP1644120A1 (de) 2006-04-12
US7776221B2 (en) 2010-08-17
US20070175830A1 (en) 2007-08-02
DE10331254A1 (de) 2005-02-10
JP4762139B2 (ja) 2011-08-31
JP2009513318A (ja) 2009-04-02
DK1644120T3 (da) 2012-03-19
DE10331254B4 (de) 2006-05-04
CA2531399C (en) 2012-01-31
ES2377733T3 (es) 2012-03-30
ATE535305T1 (de) 2011-12-15
WO2005005049A1 (de) 2005-01-20

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