Abstract
A device for the reversible immobilization of biomolecules includes a container which can be filled with a liquid containing biomolecules and has an opening and a valve. The valve can be opened and closed by a closing mechanism for the controllable drainage of the liquid. Magnetic particles, to which the biomolecules can be immobilized can be arranged freely movable in the container. A magnet for fixing the magnetic particles in the container is arranged at the container, and the liquid can be removed from the container through the opening in the open state of the valve.
Claims
1. A device for the reversible immobilization of biomolecules, wherein the device comprising: comprises a container configured to be filled with a liquid containing biomolecules and having an opening and a valve, the valve configured to be opened and closed by a closing mechanism to enable controllable drainage of the liquid; wherein magnetic particles, to which the biomolecules are capable of being immobilized, the magnetic particles arranged to be freely movable in the container; and a magnet for fixing the magnetic particles in the container is arranged at the container, the liquid being removable from the container through the opening in an open state of the valve.
2. The device according to claim 1, wherein the closing mechanism is a pressure changer, and the pressure changer is configured to change a pressure on the liquid such that a retention force of the valve is capable of being overcome by the pressure to open the valve.
3. The device according to claim 1, wherein the closing mechanism is configured to change a polarity or a viscosity or a surface tension of the liquid in the container, so that a retention force of the valve is capable of being overcome to open the valve.
4. The device according to claim 2, wherein the pressure changer is a hydrostatic pressure changer, and the hydrostatic pressure changer is configured to increase a hydrostatic pressure of the liquid by adding additional liquid into the container such that a retention force of the valve is capable of being overcome by the hydrostatic pressure to open the valve.
5. The device according to claim 2, wherein the pressure changer is configured to change the air pressure above the liquid or at the opening.
6. The device according to claim 1, wherein the valve is arranged at the opening.
7. The device according to claim 1, wherein the valve is a capillary or filter or a film or a collecting container.
8. The device according to claim 1, wherein the opening is configured to be closed with a bead which is floatable on the liquid.
9. The device according to claim 1, further comprising a measuring instrument arranged at the opening or in the container, and being configured to carry out a measurement on a drop hanging at the opening or in the container, respectively.
10. The device according to claim 1, wherein the device comprises a mixer.
11. The device according to claim 10, wherein the mixer is a modifiable magnetic field or a magnetically movable solid body.
12. The device according to claim 1, wherein the container is a multiwell plate.
13. The device according to claim 12, wherein the wells comprise a plurality of valves or openings.
14. The device according to claim 13, wherein the closing mechanism is a pressure changer, and the pressure changer is a pressure chamber arrangement so that each well of the wells is configured to be individually applied with pressure.
15. The device according to claim 1, wherein the device is one of a plurality of devices connected in series.
16. A method for the reversible immobilization of biomolecules, the method comprising: operating the device according to claim 1.
17. A method for the reversible immobilization of biomolecules, the method comprising: arranging magnetic particles and a liquid containing biomolecules in a container; bonding of the biomolecules to the magnetic particles; fixing the magnetic particles with a magnet in the container; removing the liquid from an opening of the container by opening a valve; dissolving the biomolecules from the magnetic particles; removing the dissolved biomolecules by opening the valve.
18. An apparatus for the automated processing of biomolecules comprising: a device according to claim 1.
19. The device according to claim 1, wherein the biomolecules are capable of being reversibly immobilized by the the magnetic particles.
20. The device according to claim 12, wherein the container is a microtiter plate.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0073] The invention will be explained in more detail hereinafter with reference to the drawings.
[0074] FIG. 1 is a schematic representation of a device for the reversible immobilization and purification of biomolecules.
[0075] FIG. 2 is a schematic representation of a further embodiment of a device for the reversible immobilization and purification of biomolecules.
[0076] FIG. 3 is a schematic representation of a further embodiment of a device for the reversible immobilization and purification of biomolecules.
[0077] FIG. 4 is a first embodiment of a valve.
[0078] FIG. 5 is a second embodiment of a valve.
[0079] FIG. 6 is a third embodiment of a valve.
[0080] FIG. 7 is a schematic representation of a further embodiment of a device for the reversible immobilization and purification of biomolecules.
DETAILED DESCRIPTION OF THE EMBODIMENTS
[0081] FIG. 1 shows a schematic representation of a device 1 for the reversible immobilization and purification of biomolecules. In this embodiment, the container is designed as multiwell plate 21. The wells 22 of the multiwell plate 21 can be filled with a liquid 6. In this embodiment, the magnetic particles 3 are arranged in the wells 22 of the multiwell plate 21 and are designed as a collection of magnetic particles. In a method for processing biomolecules, a liquid 6 with the biomolecules to be processed together with the reagents required for this purpose would be located in the wells 22 of the multiwell plate 21. The biomolecules, which are located in the liquid 6, can be reversibly attached to the magnetic particles 3 (i.e. they can be immobilized). The desired biomolecules can be selectively bonded to the magnetic particles. The non-bonded impurities are then removed via the opening. In addition, the biomolecules can be extended e.g. at the surface of the magnetic particles 3 (e.g. by PCR). After a completed reaction, any impurities which have been formed during the reaction or which have not completely reacted, and which are present in the liquid 6 must be removed. For this purpose, a pressure p generated by a pressure changer, which here is designed as a pressure chamber arrangement 41 (here device generating a pressure p), can overcome the retention force of the valve 20 by exerting a pressure on the liquid 6 (not shown here) located in the wells. In this way, the liquid 6 can be removed from the multi well plate 21, while the biomolecules remain on the surface of the magnetic particles 3. The magnetic particles are held in the well 22 of the multiwell plate 21 by a magnet 5.
[0082] FIG. 2 shows a schematic representation of a further embodiment of a device 1 for the reversible immobilization and purification of biomolecules. in this device 1, a floatable bead 7 is arranged in the container 2, 21 in the well 22. In condition A, in which there is no liquid 6 in the container 2, 21, the floating bead 7 closes the opening 23 and the valve 20. The valve 20 can be, for example, a capillary in which the liquid 6 is held by the capillary forces.
[0083] In the embodiment in which the container 2, 21 is designed as a multiwell plate 21, and in which several wells 22 are arranged next to each other, a pressure drop can thus be prevented when emptying the wells 22 by applying a pressure p (not shown here) generated by the pressure changer (here the device generating a pressure p). The pressure drop occurs when one well of the multiwell plate 21 is already empty, i.e. is in condition A, while other wells 22 of the multiwell plate 21 are still filled with liquid 6, i.e. are in condition B. The pressure drop can be prevented by closing the opening 23 of a well 22, which is in condition A, by the floatable bead 7.
[0084] In condition B, in which the well 22 is filled with liquid, the floatable bead 7 floats on the surface of the liquid 6 and thus allows the liquid 6 to be removed from the opening 23 by applying a pressure p (not shown here). In condition B, the liquid 6 is held by the valve 20 in the well 22 of the container 2, 21 and cannot drain through the opening 23. The liquid 6 can drain from the opening 23 only when the valve 20 is opened.
[0085] A floatable bead can he used, for example, in a device as shown in FIG. 1.
[0086] FIG. 3 shows a schematic representation of a further embodiment of a device for the reversible immobilization and purification of biomolecules. In this embodiment, a liquid 6 with magnetic particles 3 is located in the container 2, 21. The liquid 6 is retained by a valve 20 in the form of a capillary 201. Furthermore, a stirring rod 81 is located in the well 22 of the container 2, 21, This stirring rod 81 is suitable for setting the liquid 6 in motion in such a way that the liquid 6 is thoroughly mixed during a reaction step. During a washing step, the liquid 6 can drain faster by applying a pressure p (not shown here) if the liquid 6 is set in motion by the stirring rod 81.
[0087] Of course, the stirring rod 81 shown in FIG. 3 can be combined with any valve 20 and the stirring rod 81 can also be designed as another magnetically movable solid body.
[0088] FIG. 4 shows a first embodiment of a valve. In this embodiment, the valve of the container 2, 21, is designed as a film 203. The opening 23 need not be a capillary but can simply be designed as a channel. Due to the film 203, the liquid 6 cannot drain through the opening 23 from the well 22 of the container 2, 21, because the liquid is held in the container by a negative pressure. Only when the film 203 is moved, when the gas volume between film and liquid 6 is compressed, i.e. when a pressure P3 is applied to the liquid, the liquid 6 can drain through the opening 23. The film 203 could be moved by a pressure changer in such a way that the film 203 causes a lowering of the film 203 in the direction of the liquid 6 by a pressure (not shown here) on the film from the side away from the liquid. In a method according to the invention, the magnetic particles 3 could be held in the well 22 by a magnet 5 in a washing step, while the liquid 6 together with impurities could drain when moving the film 203 (magnetic particles 3 and magnet 5 see FIG. 1). Of course, a valve according to FIG. 4 can be combined with a device 1 according to FIG. 1, as well as with a floatable bead 7 according to FIG. 2 and a stirring rod 81 according to FIG. 3.
[0089] FIG. 5 shows a second embodiment of a valve. In this embodiment, the valve of the container 2, 21 is designed as a collecting container 204. An excess pressure P1 is generated in the collecting container 204 in such a way that the liquid 6 cannot drain of the well 22 of the container 2, 21 through the opening 23. Only when the container 2, 21 and the collecting container 204 are pulled apart, when the excess pressure P1 adapts to the ambient pressure P2, the liquid 6 can drain through the opening 23. In a method according to the invention, the magnetic particles 3 could be held in the well 22 by a magnet 5 in a washing step, while the liquid 6 together with impurities can drain when the container 2, 21 and the collection container 204 are pulled apart (magnetic particles 3 and magnet 5 see FIG. 1). In this embodiment, a pressure changer would correspond to a device for pulling apart the container 2, 21 and the collecting container 204, as this changes the excess pressure P1 to the ambient pressure P2, allowing the liquid 6 to drain. Of course, a valve according to FIG. 5 can be combined with a device 1 according to FIG. 1, as well as with a stirring rod 81 according to FIG. 3. In addition, it is possible that a pressure change is implied differently. For example, the pressure change can be caused by a closable opening, which is arranged on the collecting container 204. FIG. 6 shows a third embodiment of a valve. in the case of the container 2, 21, the valve is designed as a filter 202. The liquid 6 is retained by the filter 202, so that the liquid 6 cannot drain through the opening 23 from the well 22 of the container 2, 21. Only when a pressure P (not shown here) is generated by a pressure changer (here rather a pressure generator), which applies the liquid 6 in such a way that the liquid 6 is pressed through the filter 202, the liquid 6 can drain through the opening 23. In a method according to the invention, the magnetic particles 3 could be held in the well 22 by a magnet 5 in a washing step, while the liquid 6 together with impurities can drain when applying with pressure. In this embodiment, a pressure changer would correspond to a device for generating pressure, since this overcomes the retention force of the 202 filter, allowing the liquid 6 to drain. Of course, a valve according to FIG. 6 can be combined with a device 1 according to FIG. 1, as well as with a stirring rod 81 according to FIG. 3.
[0090] FIG. 7 shows a schematic representation of a further embodiment of a device for the reversible immobilization and purification of biomolecules. This embodiment shows a series connection of several devices. In this way, a liquid 6 can be transferred from an upper container 2, 21 to a lower container 2, 21 by actuating the valve 20 to transfer the liquid from one opening 23 to the next container 2, 21. The valves 20 of the different containers can all be the same or all different or partially different, For example, a first valve 205 could be a capillary 201, while a second valve 206 is a filter. But 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 second capillaries 2012, 2013 can be of different length and/or thickness, whereby a different residence time of the liquid 6 is achieved in each container 2, 21, Of course, a series connection according to FIG. 7 can be combined with a device 1 according to FIG. 1, as well as a floatable bead 7 according to FIG. 2 and a stirring rod 81 according to FIG. 3. In addition, with a series connection, various process steps can be carried out at each level of the device,
