EP2192987A1 - Apparatus and method for the treatment of liquids with magnetic particles - Google Patents
Apparatus and method for the treatment of liquids with magnetic particlesInfo
- Publication number
- EP2192987A1 EP2192987A1 EP08804473A EP08804473A EP2192987A1 EP 2192987 A1 EP2192987 A1 EP 2192987A1 EP 08804473 A EP08804473 A EP 08804473A EP 08804473 A EP08804473 A EP 08804473A EP 2192987 A1 EP2192987 A1 EP 2192987A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- magnetic particles
- central element
- magnetic
- ratio
- liquid
- 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.)
- Granted
Links
- 239000006249 magnetic particle Substances 0.000 title claims abstract description 102
- 239000007788 liquid Substances 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 28
- 230000005291 magnetic effect Effects 0.000 claims description 22
- 238000000926 separation method Methods 0.000 claims description 13
- 239000000463 material Substances 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 239000003302 ferromagnetic material Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 230000005415 magnetization Effects 0.000 claims description 2
- 239000002907 paramagnetic material Substances 0.000 claims description 2
- 239000002902 ferrimagnetic material Substances 0.000 claims 1
- 230000003993 interaction Effects 0.000 claims 1
- 239000002245 particle Substances 0.000 abstract description 2
- 238000000265 homogenisation Methods 0.000 description 13
- 108020004707 nucleic acids Proteins 0.000 description 8
- 102000039446 nucleic acids Human genes 0.000 description 8
- 150000007523 nucleic acids Chemical class 0.000 description 8
- 238000003756 stirring Methods 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 238000010828 elution Methods 0.000 description 6
- 239000013076 target substance Substances 0.000 description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 238000007885 magnetic separation Methods 0.000 description 4
- 239000008188 pellet Substances 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000009739 binding Methods 0.000 description 3
- 230000002349 favourable effect Effects 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000013049 sediment Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000027455 binding Effects 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000011534 wash buffer Substances 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 108091093037 Peptide nucleic acid Proteins 0.000 description 1
- -1 RNA Chemical class 0.000 description 1
- 108020004688 Small Nuclear RNA Proteins 0.000 description 1
- 102000039471 Small Nuclear RNA Human genes 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 TeflonĀ® Polymers 0.000 description 1
- 108020004566 Transfer RNA Proteins 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 238000012443 analytical study Methods 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 230000002051 biphasic effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
- 238000010908 decantation Methods 0.000 description 1
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 description 1
- SHIBSTMRCDJXLN-KCZCNTNESA-N digoxigenin Chemical compound C1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)C[C@H]3O)C)=CC(=O)OC1 SHIBSTMRCDJXLN-KCZCNTNESA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 108091070501 miRNA Proteins 0.000 description 1
- 239000002679 microRNA Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000000284 resting effect Effects 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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
- B03C1/288—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
- B03C1/025—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
-
- 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
- B03C1/0335—Component parts; Auxiliary operations characterised by the magnetic circuit using coils
-
- 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
- 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 and a method for treating liquids with magnetic particles.
- the device and the method are suitable, for example, for applications in biochemistry, clinical chemistry, molecular biology, microbiology, medical diagnostics or forensic medicine.
- the magnetic particles are immobilized by applying magnetic forces or a magnetic field, for example by means of a permanent magnet, at one point. This accumulation of magnetic particles is also referred to as pellet or magnetic sediment. Subsequently, the liquid supernatant is separated, for example, by suction or decantation and discarded. Since the magnetic particles are immobilized by the magnetic forces, it is largely prevented that magnetic particles are separated together with the supernatant.
- the immobilized magnetic particles are then resuspended.
- an elution liquid or an elution buffer is used to enrich the bound target substances.
- the bond between the target Substance and the magnetic particles is dissolved and released the target substance molecules from the magnetic particles.
- the target substance molecules can then be separated together with the elution liquid, while the magnetic particles are immobilized by the action of a magnetic field.
- the target substance molecules can not only be enriched, but also concentrated.
- one or more washing steps can be carried out.
- US 2001/0022948 describes an apparatus in which a magnetic rod is immersed in a first reaction vessel containing magnetic particles suspended in liquid.
- EP 0 965 842 discloses a device in which the magnetic particles, together with the liquid in which they are suspended, are drawn up in a pipette.
- the pipette tip has a special separation area, which can be acted upon by a magnet with a magnetic field.
- the magnetic particles as a pellet or magnetic sediment immobilized on the inside of the pipette tip.
- the aspirated liquid is removed from the pipette tip by the pipetting function of the device.
- EP 015 905 520 Another principle for the separation of magnetic particles is described in EP 015 905 520.
- the magnetic particles remain in the same reaction vessel while the liquid in this vessel is exchanged.
- the magnetic sediments can be immobilized at a desired height on the side wall of the reaction vessel for adaptation to a respective process step. This is done by providing magnets which are arranged on different arms of a rotatably mounted carrier at a different distance from the axis of rotation. By rotating the carrier, a particular arm can be brought into the vicinity of the side wall of the reaction vessel, and thus a specific magnet. At this point, the magnetic particles are then immobilized as a pellet.
- the present invention has for its object to overcome the described, resulting from the prior art disadvantages and in particular for a wide range of applications to provide a device and a method in which a treatment of liquids with magnetic particles in a simple manner possible is.
- an apparatus for treating liquids with magnetic particles comprising a plurality of first, disposed in the liquid magnetic particles and at least one, arranged in the liquid, preferably rod, dumbbell and / or ellipsoidal shaped magnetic and / or magnetizable central element in which the ratio of the longest diameter d2 of the at least one central element to the ratio of the average diameter d1 of the magnetic particles is at least
- central element is understood to mean, in particular, any article which is capable of being subjected to magnetic field action, possibly under the action of a further, āexternalā magnet (as described below) - to bind at rest in at least the majority of the magnetic particles.
- the at least one central element comprises a magnet, preferably a permanent magnet; According to an alternative preferred embodiment of the invention, the at least one central element comprises a magnetizable material, e.g. Iron.
- liquids is understood to mean, but is not limited to, aqueous solutions, suspensions and / or biphasic emulsions with water as a phase containing biomolecules.
- treating in the sense of the present invention is understood in particular to mean that certain biomolecules can attach to the magnetic particles in a separation step, but the present invention is expressly not limited thereto.
- diameter of the magnetic particles is meant in particular, when the magnetic particles are not spherical or substantially spherical, the respective longest diameter of the magnetic particles.
- average diameter is meant in particular the arithmetic mean of the diameters of the magnetic particles, which can be measured randomly, in particular (but not limited thereto).
- Such a device provides at least one of the following advantages for a wide range of applications within the present inventions: Characterized in that at least one central element is provided, a homogenization of the magnetic particles as well as a separation of the magnetic particles from the solution in a simple manner possible, as described below, inter alia.
- the device allows use in "closed" systems, this is a preferred embodiment of the invention in that it requires no further means (such as a bar magnet, etc.) which dip directly into the liquid and thus constitute a potential source of contamination. It results in a very fast and easy for most applications
- the ratio of the longest diameter d2 of the at least one central element to the ratio of the average diameter d1 of the magnetic particles d2 (mm) is> 50 * dl (mm), more preferably d2 (mm)> 100 * dl (mm Further, d2 (mm)> 200 * dl (mm), and most preferably d2 (mm)> 300 * dl (mm).
- the ratio of the volume V2 is the at least one central element to the ratio of the average volume Vl of the magnetic particles V2 (mm 3)> 100 * V1 (mm 3), more preferably, V2 (mm 3)> 1000 * V1 (mm 3 ) and most preferably V2 (mm 3 )> 10 5 * V1 (mm 3 ).
- the number of magnetic particles per central element is> 10 4 to ā 10 8 , preferably> 5 ā 10 5 to ā 5 ā 10 6 .
- the magnetic particles comprise a material selected from the group of paramagnetic materials, superparamagnetic materials, ferromagnetic materials, ferromagnetic materials and mixtures thereof.
- the average saturation magnetization of the magnetic particles is> 1 Am 2 / kg and 250 AmVkg, preferably> 10 AmVkg and 240 Am 2 / kg, and most preferably> 20 AmVkg and 235 AmVkg.
- the at least one central element is rod, dumbbell and / or ellipsoidal and the ratio of the longest diameter a to the ratio of the shortest diameter b is from
- the at least one central element is rod, dumbbell and / or ellipsoidal in shape and the ratio of the longest diameter a to the ratio of the shortest diameter b is from a / b> 1.5 to a / b ā 8, preferably a / b> 2 to a / b ā 5.
- the magnetic particles and the at least one central element are arranged in a closed vessel.
- the added volume V m of the magnetic particles and that of the at least one central element are> 0.25% to ā 50% of the total volume V G of the vessel. This has proven to be beneficial for many applications.
- the added volume V m of the magnetic particles and that of the at least one central element are> 0.5% to ā 20%, more preferably> 1% to ā 15% of the total volume V G of the vessel.
- the device according to the invention further comprises at least one external magnet, which is designed to interact with the at least one central element.
- the central element is a permanent magnet
- the ratio of the magnet strength H3 of the at least one external magnet to the magnet strength H 2 of the at least one central element H 3 > 1.5 * H 2 to H 3 ā 8 * H 2 , even more preferably H 3 > 2 * H 2 to H 3 ā 5 * H 2 .
- the at least one external magnet is and / or comprises an electromagnet (s) which is operated to homogenize the magnetic particles under alternating voltage.
- the central element is then preferably a (permanent) magnet.
- the at least one external magnet is and / or comprises a permanent magnet (s).
- the present invention also relates to a method of treating liquids with magnetic particles comprising a plurality of first magnetic particles arranged in the liquid and at least one in the liquid, preferably rod, dumbbell and / or ellipsoidal - Shaped trained magnetic or magnetizable central element, comprising the steps
- step a) comprises a resuspension of the magnetic particles in the liquid.
- step a) prior to step a) the magnetic particles were at least partially, preferably almost completely, attached to the at least one central element and step a) takes place by the action of an external force on the at least one central element.
- step b) is supported by means of a further, external permanent magnet.
- a further, external permanent magnet is brought into the vicinity of the vessel in which the magnetic particles and the at least one central element are located.
- the method according to the invention comprises a device according to the invention.
- the present invention also relates to the use of a device according to the invention and / or a method according to the invention for the at least partial separation of biomolecules from / in a preferably aqueous solution.
- biomolecules includes, but is not limited to, all biomolecules, such as lipids, carbohydrates, metabolites, metabolites, all types of nucleic acids, all types of peptides and proteins, and also substituted or functionalized peptides and / or proteins understood.
- biomolecules are understood to mean, but is not limited to, all naturally occurring or artificially introduced molecules in biological samples.
- the device according to the invention and / or the method according to the invention is used for the at least partial removal of nucleic acids from / in a preferably aqueous solution.
- nucleic acid in the context of the present invention particularly, but not limited to, natural, preferably isolated, linear, branched or circular nucleic acids such as RNA, in particular mRNA, siRNA, miRNA, snRNA, tRNA, hnRNA or ribozymes, DNA and the like, synthetic or modified nucleic acids, for example oligonucleotides, in particular primers, probes or standards used for the PCR, nucleic acids labeled with digoxigenin, biotin or fluorescent dyes or so-called PNAs ("peptide nucleic acids").
- RNA in particular mRNA, siRNA, miRNA, snRNA, tRNA, hnRNA or ribozymes, DNA and the like
- synthetic or modified nucleic acids for example oligonucleotides, in particular primers, probes or standards used for the PCR, nucleic acids labeled with digoxigenin, biotin or fluorescent dyes or so-called PNAs ("peptide nucle
- FIG. 1 shows a very schematic view of a device according to the invention according to an embodiment of the invention prior to "homogenization" of the magnetic particles;
- Fig. 2 shows the device of Fig. 1 after "homogenization"
- FIG. 3 shows a UV curve of a DNA elution solution after carrying out a preparation of genomic DNA according to Example I.
- FIG. 3 shows a UV curve of a DNA elution solution after carrying out a preparation of genomic DNA according to Example I.
- FIG. 1 shows a very schematic view of a device according to the invention according to an embodiment. It should be noted that FIGS. 1 and 2 are highly schematic, and in most of the inventors' applications, the actual ratios (whether size ratios such as the number of magnetic particles) will be different.
- the device comprises a plurality of first magnetic particles 10, which are deposited in the "resting state" on a central element 20.
- Magnetic particles 10 and central magnet 20 are arranged in a (preferably closed) vessel 100, which optionally via (schematically indicated by dashed lines) inlets and outlets 110 or 120.
- the vessel 100 is preferably filled with a liquid 150 so high that the magnetic particles 10 and the central element 20 are in the liquid.
- the central member 20 is a permanent magnet; however, this is not limiting.
- the central element 20 may also contain a magnetizable material such as iron.
- Another embodiment for moving the at least one central element is a one-dimensional oscillating movement. Due to the effect of a magnetic field that moves back and forth along a line, the at least one central element is "hewn" alternately against the opposite vessel walls, as a result of which the magnetic particles are also shaken off effectively by the central element 20.
- a shaking movement of the at least one central element can also take place by means of an electromagnet when it is operated under alternating voltage and the magnetic field polarity changes alternately, in that respect this also constitutes a preferred embodiment of the invention. Changing the operating mode to direct current thus finds a magnetic separation instead of.
- the magnetic particles 10 re-attach to the central element 20, so that (substantially) the state of FIG. 1 is reached again.
- the liquid 150 may now be e.g. be removed from the vessel or other reagents may be added, depending on the specific application.
- Example I Preparation of genomic DNA from 5 ml of whole blood
- Buffer AL brand product from QIAGEN
- QIAGEN Proteinase K
- the supernatant was removed, followed by air drying of the magnetic particles for 20 min.
- the UV curve of the supernatant can be seen in FIG. Based on the UV spectrum, the yield can be estimated, which was approximately quantitative in Example 1 (about 170 micrograms of genomic DNA from 5 ml of whole blood).
Landscapes
- Physical Or Chemical Processes And Apparatus (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102007045474A DE102007045474A1 (en) | 2007-09-21 | 2007-09-21 | Apparatus and method for treating liquids with magnetic particles |
PCT/EP2008/062539 WO2009040312A1 (en) | 2007-09-21 | 2008-09-19 | Apparatus and method for the treatment of liquids with magnetic particles |
Publications (2)
Publication Number | Publication Date |
---|---|
EP2192987A1 true EP2192987A1 (en) | 2010-06-09 |
EP2192987B1 EP2192987B1 (en) | 2020-04-22 |
Family
ID=40350080
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP08804473.0A Active EP2192987B1 (en) | 2007-09-21 | 2008-09-19 | Apparatus and method for the treatment of liquids with magnetic particles |
Country Status (5)
Country | Link |
---|---|
US (1) | US8361326B2 (en) |
EP (1) | EP2192987B1 (en) |
JP (1) | JP5336495B2 (en) |
DE (1) | DE102007045474A1 (en) |
WO (1) | WO2009040312A1 (en) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2574931B1 (en) | 2011-09-29 | 2017-03-22 | Qiagen GmbH | Dry composition comprising a control dye |
EP2766493B1 (en) | 2011-10-11 | 2019-12-18 | Qiagen GmbH | Sample processing method and sample processing cartridge |
JP2015159733A (en) * | 2014-02-26 | 2015-09-07 | ć»ć¤ć³ć¼ćØćć½ć³ę Ŗå¼ä¼ē¤¾ | Lyophilizate of substance binding solid-phase carrier, vessel to bind substance in substance-containing liquid to substance binding solid-phase carrier, and manufacturing method of lyophilizate containing substance binding solid-phase carrier |
US10711265B2 (en) | 2015-06-01 | 2020-07-14 | Qiagen Gmbh | Electrophoresis assisted method for purifying a target nucleic acid using a delayed elution approach |
EP3303582A1 (en) | 2015-06-01 | 2018-04-11 | Qiagen GmbH | Electrophoresis assisted method and device for purifying a charged target molecule from a sample |
KR101834828B1 (en) | 2017-06-02 | 2018-03-07 | ķźµź³¼ķźø°ģ ģ | Bacteria concentration method using magnetic force |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
NL8400855A (en) | 1984-03-16 | 1985-10-16 | Wagemans Maastricht B V | SPRING HINGE. |
JPH04102062A (en) * | 1990-08-22 | 1992-04-03 | Hitachi Ltd | Method for measuring particle immunity |
US5897783A (en) * | 1992-09-24 | 1999-04-27 | Amersham International Plc | Magnetic separation method |
DE4423878A1 (en) | 1994-07-07 | 1996-01-11 | Boehringer Mannheim Gmbh | Device and method for separating magnetic microparticles |
FI944939A0 (en) * | 1994-10-20 | 1994-10-20 | Labsystems Oy | Foerfarande Foer separering av partiklar |
FI944940A0 (en) | 1994-10-20 | 1994-10-20 | Labsystems Oy | Tvaofasigt separeringsfoerfarande |
WO1997044671A1 (en) | 1996-05-20 | 1997-11-27 | Precision System Science Co., Ltd. | Method and apparatus for controlling magnetic particles by pipetting machine |
EP1712921A2 (en) | 1997-09-29 | 2006-10-18 | F.Hoffmann-La Roche Ag | Apparatus for separating magnetic particles |
FI102906B (en) | 1998-02-23 | 1999-03-15 | Bio Nobile Oy | Procedure and means for transporting a substance |
FI20000583A0 (en) | 2000-03-14 | 2000-03-14 | Labsystems Oy | Dish and rod |
WO2003072531A1 (en) * | 2002-02-01 | 2003-09-04 | Exportech Company, Inc. | Process and apparatus for recovery of magnetic particles in a slurry phase reactor |
FI120863B (en) * | 2002-10-18 | 2010-04-15 | Biocontrol Systems Inc | Magnetic transfer method and microparticle transfer device |
DE10331254B4 (en) * | 2003-07-10 | 2006-05-04 | Chemagen Biopolymer-Technologie Aktiengesellschaft | Apparatus and method for separating magnetic or magnetizable particles from a liquid |
FI20051248L (en) | 2005-12-02 | 2007-06-03 | Bio Nobile Oy | Enrichment unit and enrichment method for biological components |
-
2007
- 2007-09-21 DE DE102007045474A patent/DE102007045474A1/en not_active Withdrawn
-
2008
- 2008-09-19 EP EP08804473.0A patent/EP2192987B1/en active Active
- 2008-09-19 US US12/676,376 patent/US8361326B2/en active Active
- 2008-09-19 WO PCT/EP2008/062539 patent/WO2009040312A1/en active Application Filing
- 2008-09-19 JP JP2010525360A patent/JP5336495B2/en active Active
Non-Patent Citations (1)
Title |
---|
See references of WO2009040312A1 * |
Also Published As
Publication number | Publication date |
---|---|
US8361326B2 (en) | 2013-01-29 |
JP2010539502A (en) | 2010-12-16 |
DE102007045474A1 (en) | 2009-04-02 |
US20100307981A1 (en) | 2010-12-09 |
WO2009040312A1 (en) | 2009-04-02 |
EP2192987B1 (en) | 2020-04-22 |
JP5336495B2 (en) | 2013-11-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP1446668B1 (en) | Device and method for treating magnetic particles | |
EP2033715B1 (en) | Method for suspending or re-suspending particles in a solution and device adapted therefor | |
EP1919625B1 (en) | Device and method for the separation of magnetic particles from a liquid | |
EP1843854B1 (en) | Device and method for the elimination of magnetic or magnetizable particles from a liquid | |
EP1644120B1 (en) | Device and method for removing magnetic or magnetisable particles from a liquid | |
EP2192987B1 (en) | Apparatus and method for the treatment of liquids with magnetic particles | |
DE60207564T2 (en) | TREATMENT METHOD FOR MAGNETIC PARTICLES AND BIOANALYSIS APPARATUS USING MAGNETIC APPLICATION | |
WO2008145587A1 (en) | Method for purifying cells, recovering cells, and transfecting cells gently | |
DE102017204267B4 (en) | METHOD OF ENRICHING CELLS FROM A SAMPLE AND THE SUBSEQUENT NUCLEIC ACID ISOLATION FROM THESE CELLS | |
WO2003009943A1 (en) | System for separating magnetically attractable particles | |
EP1333932B1 (en) | Method for separating a dispersed or dissolved substance and magnet separator | |
DE3200988A1 (en) | METHOD AND DEVICE FOR SEPARATING ORGANIC SUBSTANCES FROM A SUSPENSION OR SOLUTION | |
DE102008057317A1 (en) | Device and method for the purification of biomolecules | |
WO2011012121A1 (en) | Method for isolating high-purity rna by means of paramagnetic microparticles and nanoparticles | |
EP3693739A1 (en) | Method and device for isolating desired cells from a sample of nonmagnetic biological material | |
EP1274745A1 (en) | Magnetic, silanised polyvinylalcohol-basedcarrier materials | |
DE112019004459T5 (en) | ENRICHMENT OF SAMPLES BY MEANS OF MAGNETIC PARTICLES IN MICROCHANNELS | |
EP3147028B1 (en) | Method and system for magnetic separation of nano-beads | |
DE102013009773B4 (en) | Device and method for increasing the binding efficiency of binding capable target structures | |
DE202021105458U1 (en) | Device for the magnetic purification of biological samples | |
WO2007000399A1 (en) | Carrier material, method for the production and use thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20100421 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MT NL NO PL PT RO SE SI SK TR |
|
AX | Request for extension of the european patent |
Extension state: AL BA MK RS |
|
DAX | Request for extension of the european patent (deleted) | ||
17Q | First examination report despatched |
Effective date: 20130605 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R079 Ref document number: 502008017067 Country of ref document: DE Free format text: PREVIOUS MAIN CLASS: B03C0001280000 Ipc: B03C0001033000 |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: GRANT OF PATENT IS INTENDED |
|
RIC1 | Information provided on ipc code assigned before grant |
Ipc: B03C 1/01 20060101ALI20191219BHEP Ipc: B03C 1/033 20060101AFI20191219BHEP Ipc: B03C 1/28 20060101ALI20191219BHEP |
|
INTG | Intention to grant announced |
Effective date: 20200113 |
|
GRAS | Grant fee paid |
Free format text: ORIGINAL CODE: EPIDOSNIGR3 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE PATENT HAS BEEN GRANTED |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MT NL NO PL PT RO SE SI SK TR |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: FG4D Free format text: NOT ENGLISH |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: EP |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R096 Ref document number: 502008017067 Country of ref document: DE |
|
REG | Reference to a national code |
Ref country code: IE Ref legal event code: FG4D Free format text: LANGUAGE OF EP DOCUMENT: GERMAN |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: REF Ref document number: 1259374 Country of ref document: AT Kind code of ref document: T Effective date: 20200515 |
|
REG | Reference to a national code |
Ref country code: LT Ref legal event code: MG4D |
|
REG | Reference to a national code |
Ref country code: NL Ref legal event code: MP Effective date: 20200422 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: NL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: LT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: PT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200824 Ref country code: SE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: IS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200822 Ref country code: FI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: NO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200722 Ref country code: GR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200723 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: LV Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: HR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: BG Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200722 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R097 Ref document number: 502008017067 Country of ref document: DE |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: ES Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: CZ Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: RO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: EE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: DK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: PL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 |
|
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
26N | No opposition filed |
Effective date: 20210125 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MC Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: PL |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 |
|
REG | Reference to a national code |
Ref country code: BE Ref legal event code: MM Effective date: 20200930 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: LU Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20200919 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: BE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20200930 Ref country code: CH Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20200930 Ref country code: IE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20200919 Ref country code: LI Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20200930 |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: MM01 Ref document number: 1259374 Country of ref document: AT Kind code of ref document: T Effective date: 20200919 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: AT Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20200919 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: TR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: MT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 Ref country code: CY Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200422 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: GB Payment date: 20230920 Year of fee payment: 16 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: FR Payment date: 20230928 Year of fee payment: 16 Ref country code: DE Payment date: 20230920 Year of fee payment: 16 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: IT Payment date: 20230927 Year of fee payment: 16 |