Classifications

• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
  • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
  • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
  • G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
  • B01L3/02—Burettes; Pipettes
  • B01L3/0241—Drop counters; Drop formers
  • B01L3/0268—Drop counters; Drop formers using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
  • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
  • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
  • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
  • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
  • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
  • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
  • B01L3/50255—Multi-well filtration
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
  • C12N15/09—Recombinant DNA-technology
  • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
  • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
  • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
  • C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2300/00—Additional constructional details
  • B01L2300/04—Closures and closing means
  • B01L2300/041—Connecting closures to device or container
  • B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2300/00—Additional constructional details
  • B01L2300/04—Closures and closing means
  • B01L2300/046—Function or devices integrated in the closure
  • B01L2300/049—Valves integrated in closure
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2300/00—Additional constructional details
  • B01L2300/06—Auxiliary integrated devices, integrated components
  • B01L2300/0627—Sensor or part of a sensor is integrated
  • B01L2300/0663—Whole sensors
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2300/00—Additional constructional details
  • B01L2300/08—Geometry, shape and general structure
  • B01L2300/0809—Geometry, shape and general structure rectangular shaped
  • B01L2300/0829—Multi-well plates; Microtitration plates
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2300/00—Additional constructional details
  • B01L2300/14—Means for pressure control
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2400/00—Moving or stopping fluids
  • B01L2400/04—Moving fluids with specific forces or mechanical means
  • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
  • B01L2400/043—Moving fluids with specific forces or mechanical means specific forces magnetic forces
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2400/00—Moving or stopping fluids
  • B01L2400/04—Moving fluids with specific forces or mechanical means
  • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
  • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2400/00—Moving or stopping fluids
  • B01L2400/04—Moving fluids with specific forces or mechanical means
  • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
  • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
  • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2400/00—Moving or stopping fluids
  • B01L2400/06—Valves, specific forms thereof
  • B01L2400/0677—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
  • B01L2400/0683—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers mechanically breaking a wall or membrane within a channel or chamber
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L2400/00—Moving or stopping fluids
  • B01L2400/06—Valves, specific forms thereof
  • B01L2400/0688—Valves, specific forms thereof surface tension valves, capillary stop, capillary break

Definitions

• the invention relates to a device for the reversible immobilization of biomolecules.
• the invention further relates to a method for the reversible immobilization of biomolecules according to the preamble of independent claim 16.
• the invention further relates to a device Apparatus for the automated processing of biomolecules comprising a device for the reversible immobilization of biomolecules according to the preamble of independent claim 18.
• DNA extraction which precipitates DNA in a nonpolar environment.
• DNA can be removed by centrifugation, e.g. after cell disruption or by electrophoretic methods.
• Biomolecules can also be synthesized and purified by immobilization on an insoluble support.
• Common substrates for immobilizing biomolecules are glass as well as other, less common substrates such as gold, platinum, oxides, semiconductors and various polymer substrates.
• Magnetic bead-based clean-up and “magnetic bead-based normalization” are widespread methods for immobilization, purification and
• the magnetic particles are typically held in the container by ring magnets which enclose a container. As a result, a solution containing impurities can be pipetted off, while the magnetic particles with the bound biomolecules remain in the container.
• the magnetic beads were developed at the Whitehead Institute in 1995 for the purification of PCR products.
• the magnetic particles are paramagnetic and may be e.g. made of polystyrene coated with iron. On the iron then various molecules can be attached with carboxyl groups. These carboxyl groups can reversibly bind the DNA molecules. This immobilizes the DNA molecules.
• Magnetic particle methods usually include the following steps. First, the PCR products are bound to the magnetic particles. Subsequently, the magnetic particles with the attached PCR products are separated from impurities (this step is realized, for example, by pipetting off the solution from the solid). Then the magnetic particles are washed with the attached PCR products. After washing, the PCR products are eluted from the magnetic particles and transferred to a new plate.
• the magnetic particle-bound nucleic acids are collected at the bottom and at the edge of the cavities and depending on the routine optimized pipetting up and down again brought into solution. Finally, the DNA or RNA are eluted for direct storage or further applications in separate tubes with lids.
• l-DOT Dispendix's “Immediate Drop on Demand Technology”
• l-DOT Dispendix's “Immediate Drop on Demand Technology”
• This system for liquid dispensing is based on a microtiter plate with so-called “wells” which have openings of a few micrometers in diameter on their underside.
• the liquid is held in the wells by capillary forces.
• a well-defined pressure pulse from above on a liquid-filled wells a drop of accurate volume is formed, which is discharged through the lower openings of the wells.
• Liquid quantities are dispensed in the nanoliter range, but a dispensing system does not provide a means of purification
• a dispensing device is also known from US 8,877,145 B2.
• a liquid is held by a capillary which has a liquid reservoir.
• a hydraulic pressure By applying a hydraulic pressure, the capillary forces are overcome and an accurate
• US Pat. No. 4,111,754 discloses a device in which a plastic structure for surface enlargement is attached in a capillary. In this capillary, the fluid is held by the capillary forces and antigens or antibodies can adhere to the plastic structure. Thus, the antigens or antibodies can be immobilized on the plastic surface. By adding washing liquid, the impurities can then be removed.
• a disadvantage of this device is that the antigens and antibodies are bound inside the capillary and not with the
• Carrier material can be ejected.
• the antigens and antibodies can only be eluted by being released from the container so mobilized again.
• the surface to which the biomolecules are attached can be adjusted only by changing the capillary, that is, by changing the device, and in a reaction, the carrier for the biomolecules can not be moved to better mixing, thereby also increasing the reaction time.
• the device described is also not compatible with all purification protocols, which makes it difficult to integrate the device into existing workflows
• Biomolecules are attached to stationary support materials. Thus, the methods known in the art are slow, costly and less efficient.
• the object of the invention is therefore an apparatus for immobilizing biomolecules by binding the biomolecules to a solid surface, a method for the reversible immobilization and purification of biomolecules by binding the biomolecules to a solid surface and an apparatus for the automated processing of biomolecules with a device for To provide immobilization of biomolecules which avoid the adverse effects known from the prior art.
• the object is achieved by a device for the reversible immobilization of biomolecules with the features of independent claim 1, by a method for the reversible immobilization of biomolecules with the
• the method may include the following steps. Magnetic particles and a liquid, in particular a liquid with reagents, are arranged in a container. Biomolecules or reagents are bound to the magnetic particles, in particular bound reversibly. The magnetic particles are fixed with a magnet in the container. The liquid, in particular the liquid with impurities, is removed from the opening of the container by opening the valve, in particular for the purification of the biomolecules. The biomolecules are, e.g. with a solvent, detached from the magnetic particles. Then the solved ones
• Biomolecules are removed by opening the valve from the container.
• the container may have a second opening.
• liquid can be supplied via this second opening.
• the valve can be controlled.
• the second opening may be on the opposite side of the container from the opening.
• the valve may be controlled via the second opening to regulate pressure on the liquid through the second opening.
• containers are used whose depressions have one (or more) openings, preferably on the underside, which are such that they have a valve function or can be controlled by a valve, thus it is possible to hold liquid in the depression or to empty the depression through the openings, wherein the magnetic particles are held by the magnet in the recess of the container.
• the biomolecules are reversibly bindable to the particles and the magnetic particles can have an enlarged surface compared to the container wall and can also be removed together with bound biomolecules from the container.
• biomolecules can be selectively bound to the surface of the magnetic particles so that only one sort of biomolecule is bound from a liquid.
• magnetic particles have the great advantage that the magnetic particles are simply transferred through a magnet (e.g., permanent or electromagnet) or through a magnetic field into the magnetic field
• the magnet is arranged so movable on the container that the
• magnetic particles during a reaction step in the container are freely movable and during a washing step, by changing the
• the magnet may be movable such that the magnet is in a first position on the container is arranged and the magnetic particles fixed and by moving the magnet to a second position on or around the container, the magnetic particles are movable.
• biomolecule DNA, RNA, nucleic acids, proteins, start sequences for biomolecules; Cells and
• a washing step in the context of the invention is a process step in which the liquid is discharged from the containers by actuation of the valve and in which the impurities are separated from magnetic particles with the attached biomolecules. Washing may also involve washing with a washing solution (water or others).
• a reaction step in the context of the invention is a process step in which the biomolecules bound to the magnetic particles
• chain extension e.g., PCR “polymerase chain reaction”
• Reagents may also be biomolecules and / or their monomers.
• an impurity is generally a substance which is not completely reacted or is not bound to the magnetic particles, the solvent, by-products and contaminations, as well as a mixture of two or more of those described above.
• impurities may also be reagents or biomolecules.
• a liquid may be a solution within the scope of the invention.
• reaction mixture of biomolecules and / or reagents and / or impurities.
• purification may correspond to the removal of the liquid, in particular the removal of a liquid after a washing step or the removal of the liquid between
• purification may also be understood to mean the normalization of biomolecules and the selection of biomolecules.
• valve according to the invention may be mechanical and / or electrical and / or magnetic device for closing and opening the valve.
• the valve according to the invention it is also conceivable within the scope of the invention for the valve according to the invention to be a capillary.
• a shutter mechanism could be a substance whose addition to the fluid changes the viscosity and / or the surface tension of that fluid. In such a closure mechanism / capillary combination, a pressure change would correspond to a reduction in surface tension and / or viscosity.
• a pressure changer may be a device for generating pressure (liquid and / or air pressure), such as a pump, a blower or a stamp.
• a pressure changer can also be a device which manipulates a film in such a way that a pressure can be applied to the liquid.
• a pressure changer may further be a device for pulling apart a container and a catching device so as to retain the liquid
• the retention force of the valve can in the invention
• Capillary force of a capillary the negative pressure generated by a film and generally a negative pressure, the surface tension and / or the
• Fluid barrier or a magnetic or mechanical force of a
• immobilization is to be understood as meaning the binding, in particular the reversible binding of the biomolecules to the magnetic particles.
• a magnetic particle also called “magnetic bead”
• a magnetic particle may be porous.
• a biomolecule may hereinafter generally be understood as meaning thiol groups and / or amino groups and / or
• a magnetic particle may in the context of the invention be a coated nickel particle or any other ferromagnetic or paramagnetic particle. Magnetic particles typically have a diameter of about 1 micrometer. In the context of the invention, about 1 micron is to be understood as meaning 0.5 to 1.5 microns, in particular 0.7 to 1.3 microns, in particular 0.9 to 1.1 microns.
• a valve may generally also be a pressure valve, a flow valve or a check valve, particularly preferably one
• a magnet may be a permanent magnet and / or an electromagnet and / or a superconductor and / or a ferromagnet and / or a paramagnet.
• a magnet can be a
• a measuring device In the context of the invention, a measuring device, a luminescence and
• inventive method are:
• Biomolecules can be selectively bound - By using particles much larger surface is created
• the device can be easily integrated into existing (manual, semi-automated or automated) workflows (can build on standard procedures for DNA purification)
• the closure mechanism may be a pressure changer, wherein by the pressure changer, a pressure on the liquid is variable so that a restraining force of the valve can be overcome by the pressure.
• the valve can be opened.
• the control of the pressure on the liquid is important to empty the depression as needed.
• the pressure can be through a pressure chamber with the upper part of the wells,
• the upper part is the part can be exercised over the pressure on the liquid.
• Microtitration plate can by independent pressure chambers (eg per well or per area of the multiwellplate a pressure chamber) individual area or wells are pressurized independently.
• a pressure chamber arrangement with independent pressure chambers can be connected to the upper part of the depression or of the container.
• the pressure difference can also be generated by creating a negative pressure on the outside of the opening.
• the upper part of the depression or of the container and / or the lower opening can be closed. Also reversible closure is conceivable for longer storage of samples or reagents in the container (possibly reversible closure to make a multiwell plate, in particular a microtiter plate PCR-enabled).
• a pressure changer When using a pressure changer, the opening of the valve corresponds to an increase in the pressure on the liquid or a generated negative pressure which acts on the liquid at the opening of the container. The valve is always closed when no liquid is removed from the container through the opening (only if there is still liquid in the container).
• a pressure changer can have the following principles: hydrostatics, capillary pressure, centrifugal force, gas pressure.
• the closure mechanism may be a hydrostatic pressure changer, wherein by the hydrostatic
• a hydrostatic pressure changer could be a liquid delivery device (for example, a washing liquid for a washing step).
• a polarity and / or viscosity and / or surface tension of the liquid in the container can be variable by the closure mechanism, so that a retention force of the valve can be overcome, and thus the valve can be opened.
• the polarity and / or viscosity and / or surface tension of the liquid can be changed, for example, by adding other liquids or substances, or by changing the pFI value.
• the closure mechanism could be used as a substance delivery device (for example, surfactants for
• the pressure changer can change the air pressure above the liquid and / or at the opening, in particular an opening arranged at the bottom of the container. So can the generation of a
• air pressure above the liquid is meant the air pressure which also acts on the liquid so that the liquid can be removed from the container.
• the valve of the device may be arranged at the opening.
• the valve may also be the opening, eg if the valve is a capillary, the opening of the capillary is also the opening for discharging the liquid.
• the valve and / or the opening can be arranged on the underside of the container.
• a recess of the device may comprise a plurality of valves and / or openings.
• the openings could also act as a kind of sieve, through which the magnetic particles do not fit, but the liquid can drain.
• the valve of the device may be formed as a capillary or as a filter or as a foil or as a collecting container.
• the capillary pressure is sufficient to prevent a spontaneous emptying of the cavities.
• the liquid can now be removed by applying a pressure pulse (through the pressure reducer) to the liquid from above so that the liquid is removed through the opening (opening the valve).
• a pressure pulse through the pressure reducer
• the valve is formed as a filter, the liquid is retained by the fluid barrier of the filter material.
• Liquid can also be removed here by a pressure pulse (by the pressure changer) is given from above the liquid, so that the pressure pulse (by the pressure changer) is given from above the liquid, so that the pressure pulse (by the pressure changer) is given from above the liquid, so that the pressure pulse (by the pressure changer) is given from above the liquid, so that the pressure pulse (by the pressure changer) is given from above the liquid, so that the pressure pulse (by the pressure changer) is given from above the liquid, so that the
• Liquid is forced through the filter (opening the valve) and removed through the opening.
• the valve is a film
• the film may be disposed over the container such that a volume of gas between the film and the liquid is trapped.
• manipulating the film eg by moving the film through a pressure generated by the pressure changer
• the gas volume between the liquid and the film can be compressed in this way a pressure is exerted on the liquid which forces the liquid out of the opening (opening of the valve).
• the opening of the device may be closable with a bead drivable on the liquid. So there is the possibility to empty the liquid through the opening and then to close the opening of the recess.
• the container of the device is a
• Multiwell plate in particular a microtitration plate, with depressions.
• the pressure changer of the device may be a pressure chamber arrangement, so that each recess can be acted upon individually with pressure.
• a measuring device can be arranged so that a measurement can be carried out on the hanging drop or on the liquid in the container.
• the device may comprise a mixer.
• the mixer may be a variable magnetic field and / or a magnetically movable solid.
• a magnetically movable solid may in this case be a stirring bar and / or magnetic stirrer, which is guided by a
• Magnetic field is set in motion.
• magnetic particles can be caused by a variable magnetic field, a movement of the magnetic particles, which also a
• devices may also be connected in series.
• the apparatus and method can be used for post-ligation purification.
• Biomolecules proposed carried out with a device for reversible immobilization, in particular for the purification of
• Biomolecules The method may include the following steps. Magnetic particles and a liquid with reagents are placed in a container. Biomolecules or reagents for the synthesis of
• Biomolecules are bound to the magnetic particles, in particular bound reversibly.
• the magnetic particles are fixed with a magnet in the container.
• the fluid with contaminants is removed from the mouth of the container by opening the valve to purify the biomolecules.
• the biomolecules are, e.g. with a solvent, detached from the magnetic particles. Then the dissolved biomolecules can be removed by opening the valve from the container.
• the method may, of course, include multiple steps in which liquids must be added and removed, as well as impurities are separated or in which the biomolecules are detached from the magnetic particles.
• the purified biomolecules can be dispensed by removing them through the opening of the device after detachment from the magnetic particles.
• the liquid can then be removed by a pressure change (depending on the type of valve).
• a pressure change depending on the type of valve.
• the invention also proposes an apparatus for the automated processing of biomolecules with a device for reversible immobilization, in particular for the purification of biomolecules.
• Fig. 1 is a schematic representation of a device for reversible
• Fig. 2 is a schematic representation of another
• Fig. 3 is a schematic representation of another
• FIG. 4 shows a first embodiment of a valve
• Fig. 5 shows a second embodiment of a valve
• Fig. 6 shows a third embodiment of a valve
• Fig. 7 is a schematic representation of another
• the container is designed as a multiwell plate 21.
• the depressions 22 of the multiwell plate 21 can be filled with a liquid 6.
• the magnetic particles 3 are disposed in the recesses 22 of the multiwell plate 21 and as a collection of magnetic particles
• a liquid 6 with the biomolecules to be processed together with the reagents required for this would be located in the depressions 22 of the multiwell plate 21.
• the biomolecules which are located in the liquid 6 can be reversibly attached to the magnetic particles 3 (ie be immobilized).
• the desired biomolecules can be selectively bound to the magnetic particles.
• the unbound impurities are then removed through the opening.
• the biomolecules may be e.g. are extended on the surface of the magnetic particles 3 (e.g., by PCR). After a reaction has taken place during the reaction or not fully reacted impurities, which are located in the liquid 6 are removed.
• a pressure changer which is designed here as a pressure chamber assembly 41 (here device which generates a pressure p)
• a pressure chamber assembly 41 here device which generates a pressure p
• the retention force of the valve 20 are overcome by a pressure on the liquid in the wells 6 (not here shown) is exercised.
• the liquid 6 can be removed from the multiwell plate 21, while the biomolecules remain on the surface of the magnetic particles 3.
• the magnetic particles are held by a magnet 5 in the recess 22 of the multiwell plate 21.
• Fig. 2 shows a schematic representation of another
• Embodiment of a device 1 for the reversible immobilization and purification of biomolecules a device 1 for the reversible immobilization and purification of biomolecules.
• a drivable bead 7 is mounted in the container 2, 21 in the recess 22.
• state A in which no liquid 6 in the container 2, 21, closes the Floating ball 7, the opening 23 and the valve 20.
• the valve 20 may be, for example, a capillary, in which the liquid 6 is held by the capillary forces.
• the container 2 21 is formed as a multiwell plate 21, in which a plurality of wells 22 are arranged side by side, thus when emptying the wells 22 by applying pressure generated by the pressure changer (here device which generates a pressure p) p (not shown here), a pressure drop can be prevented.
• the pressure changer here device which generates a pressure p
• State B are located.
• the pressure drop can be prevented by closing the opening 23 of a depression 22, which is in the state A, by the drivable pellet 7.
• a drivable bead may e.g. be used in a device according to Fig. 1.
• Fig. 3 shows a schematic representation of another
• Embodiment of a device for the reversible immobilization and purification of biomolecules in the container 2, 21 is a liquid 6 with magnetic particles 3.
• Liquid 6 is passed through a valve 20 in the form of a capillary 201
• a Stirrer 81 is located in the recess 22 of the container 2, 21 .
• This stirring rod 81 is adapted to the liquid 6 to set in motion such that the liquid 6 at a
• the liquid 6 can at a
• Washing step by applying a pressure p drain faster when the liquid 6 by the stirring rod 81 in
• stir bar 81 shown in Fig. 3 may be combined with any valve 20, and the stir bar 81 may be formed as another magnetically movable solid.
• Fig. 4 shows a first embodiment of a valve.
• the valve is designed as a film 203.
• the opening 23 need not be a capillary, but may simply be configured as a channel.
• the liquid 6 can not flow through the opening 23 from the recess 22 of the container 2, 21, since the liquid is held by a negative pressure in the container. Only when moving the film 203, when the gas volume between the film and liquid 6 is compressed, that is, a pressure P3 is exerted on the liquid, the liquid 6 can flow through the opening 23.
• the film 203 could be moved by a pressure changer, so that the film 203 by a pressure (not shown here) on the film from the liquid-side facing, a lowering of the film 203 in the direction of the liquid 6 causes.
• the magnetic particles 3 could be replaced by a
• Magnets 5 are held in the recess 22, while the liquid 6 can flow together with impurities when moving the film 203
• FIG. 4 Magnetic particles 3 and magnet 5, see Fig. 1.
• a valve according to Fig. 4 of course, with a device 1 according to Fig. 1, and a drivable beads 7 according to FIG. 2 and a stirring rod 81 according to FIG. 3 are combined.
• Fig. 5 shows a second embodiment of a valve.
• the valve is designed as a collecting container 204.
• an overpressure P1 is generated in such a way that the liquid 6 can not flow out of the recess 22 of the container 2, 21 through the opening 23. Only when pulling apart the container 2, 21 and the collecting container 204, when the pressure P1 adjusts to the ambient pressure P2, the liquid 6 can flow through the opening 23.
• the pressure P1 adjusts to the ambient pressure P2
• the liquid 6 can flow through the opening 23.
• inventive method could in a washing step the
• a pressure changer would correspond to a device for pulling apart the container 2, 21 and the collecting container 204, since the overpressure P1 is thus changed to the ambient pressure P2, whereby the liquid 6 can flow off.
• a valve according to FIG. 5 can, of course, be combined with a device 1 according to FIG. 1, as well as a stirring rod 81 according to FIG.
• a pressure change is also implied differently.
• the valve according to FIG. 5 can, of course, be combined with a device 1 according to FIG. 1, as well as a stirring rod 81 according to FIG.
• a pressure change is also implied differently.
• FIG. 6 shows a third embodiment of a valve.
• the valve is designed as a filter 202.
• the filter 202 Through the filter 202, the liquid is the sixth
• a pressure changer would correspond to a device for generating pressure, as this will overcome the retention force of the filter 202, allowing the liquid 6 to drain off.
• a valve according to FIG. 6 can, of course, be combined with a device 1 according to FIG. 1, as well as a stirring rod 81 according to FIG.
• Fig. 7 shows a schematic representation of another
• Embodiment of a device for the reversible immobilization and purification of biomolecules This embodiment shows a
• a liquid 6 can be transferred from an upper container 2, 21 to a lower container 2, 21 by passing the liquid from one opening 23 into the next container 2, 21 by actuation of the valve 20.
• the valves 20 of the various containers may all be the same or all different or partially different. So could e.g. a first valve 205 may be a capillary 201, while a second valve 206 is a filter. However, it would also be conceivable that a first valve 205 is a first capillary 2013, while a second valve 206 is a second capillary 2012. Thus, the first and the second capillary 2012, 2013 may be different lengths and / or thick, whereby in each container 2, 21 a different residence time of the liquid 6 is achieved.
• a series circuit according to Fig. 7 can of course with a

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• 2017
  • 2017-11-17 CA CA3081119A patent/CA3081119A1/en active Pending
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