DEVICE FOR PREPARING A DNA PRODUCT BY MEANS OF CAPILLARY POLYMERASE CHAIN REACTION

Abstract

The present invention relates to a device for preparing a DNA product by means of Capillary Polymerase Chain Reaction (PCR); a manufacturing device for a pharmaceutical product; a manufacturing module for a pharmaceutical product; a method for preparing a DNA product by means of Capillary Polymerase Chain Reaction; the use of the device for preparing a DNA product by means of Polymerase Chain Reaction; and the use of the device for a production of a pharmaceutical product. The device for preparing a DNA product by means of Capillary Polymerase Chain Reaction comprises a tube for guiding a PCR liquid, a first compartment, and at least second compartment. The tube is wound at least from the first compartment to the second compartment and from the second compartment to the first compartment, wherein the tube wound from compartment to compartment forms a helical stack of tube windings. The first compartment and the second compartment each comprises means to adjust the temperature in the compartment to prepare the DNA product based on the PCR liquid, wherein the first compartment is configured to provide a temperature for denaturation and the second compartment is configured to provide a temperature for annealing and/or elongation.

Claims

1. A device (1) for preparing a DNA product by means of Capillary Polymerase Chain Reaction (PCR), comprising: a tube (6) for guiding a PCR liquid, a first compartment (7), and at least a second compartment (8), wherein the tube (6) is wound at least from the first compartment (7) to the second compartment (8) and from the second compartment (8) to the first compartment (7), wherein the tube (6) wound from compartment to compartment forms a helical stack of tube windings, and wherein the first compartment (7) and the second compartment (8) each comprises means to adjust the temperature in the compartment to prepare the DNA product based on the PCR liquid, wherein the first compartment (7) is configured to provide a temperature for denaturation and the second compartment (8) is configured to provide a temperature for annealing and/or elongation.

2. The device (1) for preparing a DNA product according to claim 1, comprising a first compartment (7), a second compartment (8), and a third compartment (15), wherein the tube (6) is wound from the first compartment (7) to the second compartment (8), from the second compartment (8) to the third compartment (15), and from the third compartment (15) to the first compartment (7), wherein the first compartment (7), the second compartment (8) and the third compartment (15) each comprises means to adjust the temperature in the compartment to prepare the DNA product based on the PCR liquid, wherein the first compartment (7) is configured to provide a temperature for denaturation, the second compartment (8) is configured to provide a temperature for annealing and the third compartment (15) is configured to provide a temperature for elongation.

3. The device (1) for preparing a DNA product according to claim 2, wherein the tube (6) is wound through the first compartment (7) and from the first compartment (7) to and through the second compartment (8), from the second compartment (8) to and through the third compartment (15), and from the third compartment (15) to the first compartment (7).

4. The device (1) for preparing a DNA product according to claim 1, wherein each tube winding represents one PCR cycle.

5. The device (1) for preparing a DNA product according to claim 1, wherein the tube (6) comprises a PCR liquid inlet (20) and a DNA product outlet (25).

6. The device (1) for preparing a DNA product according to claim 5, wherein the PCR liquid inlet (20) is connectable to a pump unit for pumping the PCR liquid through the tube (6).

7. The device (1) for preparing a DNA product according to claim 5, wherein the DNA product outlet (25) is for a continuous preparation of the DNA product or wherein the DNA product outlet (25) is a closable DNA product outlet (25) for a discontinuous preparation of the DNA product.

8. The device (1) for preparing a DNA product according claim 1, wherein septum walls (14) are arranged between the compartments to insulate the compartments from each other.

9. The device (1) for preparing a DNA product according to claim 8, wherein the septum walls (14) comprise through holes for the tube (6) to extend from one compartment to the next.

10. The device (1) for preparing a DNA product according to claim 1, wherein each compartment comprises a heating unit (31) and optionally a cooling unit (32, 33) as the means to adjust the temperature in each compartment.

11. The device (1) for preparing a DNA product according to claim 1, wherein each compartment comprises a thermal medium inlet (12) and a thermal medium outlet (13) as the means to adjust the temperature in each compartment, namely to provide a separate thermal medium flow through the first compartment (7), the second compartment (8) and the optional third compartment (15) to adjust a separate temperature in the first compartment (7), the second compartment (8) and the optional third compartment (15).

12. The device (1) for preparing a DNA product according to claim 11, wherein the thermal medium inlet (12) and the thermal medium outlet (13) are arranged within each compartment to provide the thermal medium flow through the compartment essentially in a counter direction to a guiding direction of the PCR liquid in the tube (6).

13. The device (1) for preparing a DNA product according to claim 11, wherein the thermal medium inlet (12) is arranged at a lower position in each of the compartments and the thermal medium outlet (13) is arranged at an upper position in each of the compartments.

14. The device (1) for preparing a DNA product according to claim 11, wherein the tube (6) extends within in each of the compartments in a corrugated manner.

15-28. (canceled)

29. A manufacturing device (10) for a pharmaceutical product comprising a device (1) for preparing a DNA product by means of Capillary Polymerase Chain Reaction according to claim 1.

30. The manufacturing device (10) for a pharmaceutical product according to claim 29, further comprising a process chamber (2) and a technical chamber (3).

31. The manufacturing device (10) for a pharmaceutical product according to claim 30, further comprising a control unit configured to control a gas flow through the process chamber (2) as a gas shower.

32. The manufacturing device (10) for a pharmaceutical product according to claim 30, wherein at least the process chamber (2), preferably the manufacturing device (10), is operable according to the requirements of the EU guidelines for good manufacturing practice (GMP) for medicinal products, annex 1.

33-42. (canceled)

43. A manufacturing module (100) for a pharmaceutical product, comprising a manufacturing device (10) for a pharmaceutical product according to claim 29 and at least one manufacturing device selected from the group consisting of a manufacturing device (10) comprising a process media supply unit and a mixing unit, a manufacturing device (10) comprising a purification unit, and a manufacturing device (10) comprising a filtration unit.

44. A method for preparing a DNA product by means of Capillary Polymerase Chain Reaction (PCR), comprising: providing a first compartment (7), a second compartment (8), and a tube (6), wherein the tube (6) is wound at least from the first compartment (7) to the second compartment (8) and from the second compartment (8) to the first compartment (7), wherein the tube (6) wound from compartment to compartment forms a helical stack of tube windings, providing means to adjust a separate temperature in each compartment, and guiding a PCR liquid through the tube (6) and thereby through the separate temperatures in each compartment to prepare the DNA product based on the PCR liquid.

45-51. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0136] The Figures shown in the following are merely illustrative and shall describe the present invention in a further way. These figures shall not be construed to limit the present invention thereto.

[0137] FIG. 1 shows a top view on a device according to the present invention for preparing a DNA product by means of Capillary Polymerase Chain Reaction.

[0138] FIG. 2 shows a side view on a device according to the present invention for preparing a DNA product by means of Capillary Polymerase Chain Reaction.

[0139] FIG. 3 shows a lateral 3D view of a wound tube.

[0140] FIG. 4 shows a lateral 3D view of a tube wound around several stands.

[0141] FIG. 5 shows a 3D view of a manufacturing device for a pharmaceutical product.

[0142] FIG. 6a shows a 3D view of a box housing the device for preparing a DNA product to be inserted into the manufacturing device for a pharmaceutical product.

[0143] FIG. 6b shows a 3D view of a box housing the device for preparing a DNA product to be inserted into the manufacturing device for a pharmaceutical product.

[0144] FIG. 7 shows schematically and exemplarily a manufacturing module for a pharmaceutical product according to the invention.

[0145] FIG. 8 shows a lateral 3D view on a device according to the present invention.

[0146] FIG. 9 shows a lateral 3D view on the top of a device according to the present invention.

[0147] FIG. 10 shows a lateral 3D view on the box housing the device to be inserted into a manufacturing device.

[0148] FIG. 11 shows a 3D view zooming into a manufacturing device with the box housing the device inserted into the manufacturing device.

[0149] FIG. 1 shows a top view on a device 1 according to the present invention for preparing a DNA product by means of Capillary Polymerase Chain Reaction. FIG. 2 shows a side view of a device 1 according to the present invention for preparing a DNA product by means of Capillary Polymerase Chain Reaction. FIG. 8 shows a lateral 3D view on the device according to the present invention. In FIG. 2, the device 1 for preparing a DNA product is mounted into a manufacturing device 10 according to the present invention for a pharmaceutical product, which will be explained in more detail in view of FIG. 5. The DNA product can be DNA to be used as API or a DNA template for enzymatic RNA in vitro transcription, where the RNA would be the API. Polymerase Chain Reaction (PCR) is a technology to synthetically amplify DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. Capillary Polymerase Chain Reaction can be understood as PCR done in a capillary or tube 6. The present device 1 for preparing a DNA product allows for a flexible and fast preparation of large quantities of a DNA product. The device 1 for preparing a DNA product may be used for a flexible and fast vaccine production, particularly a DNA vaccine production or production of a DNA template for enzymatic RNA in vitro transcription. The present device 1 may be in particular used for preparing DNA templates for mRNA-based vaccines e.g. during infectious disease epidemics and pandemics.

[0150] As shown in FIG. 1 and FIG. 8, the device 1 for preparing a DNA product comprises a tube 6, a first compartment 7, a second compartment 8, and a third compartment 15. The tube 6 or capillary has a very small diameter, which can be in a range of about 0.5 mm to about 50 mm, preferably about 0.5 mm to about 1.5 mm, more preferably about 0.75 to about 1.25 mm. The tube 6 is wound from the first compartment 7 to the second compartment 8 and from the second compartment 8 to the third compartment 15. It then comes back to the first compartment 7. A PCR liquid can be filled into the tube 6 and thereby be guided through the three compartments. As shown in FIGS. 2 and 8, the compartments can be understood as independent basins, e.g. to be filled by a thermal medium (see FIG. 2) or with a thermal solid (see FIG. 8). The basins are here at least partially, preferably completely surrounded by a thermal insulation, which may be the septum wall 14 (see FIG. 1) or the inner-compartment wall 30 (see FIG. 8), optionally in combination with the septum wall 14.

[0151] The tube 6 is wound to form a helical stack of tube windings, which can be understood as a helix, spiral or coil of winding when seen in a lateral view. FIG. 3 shows a lateral 3D view of a wound tube 6. There can be several layers of windings, e.g. 30. The helical extension or stacking direction of the tube 6 extends in a z-direction essentially perpendicular to an x, y plane formed by a bottom of the device 1 for preparing a DNA product or a plane formed by the floor.

[0152] Back to FIGS. 1 and 8, the tube 6 is not only wound from the first compartment 7 to the second compartment 8 and from the second compartment 8 to the third compartment 15, but also further from the third compartment 15 to the first compartment 7, from the first compartment 7 to the second compartment 8, and from the second compartment 8 to the third compartment 15, etc. Each tube winding represents a PCR cycle.

[0153] The tube 6 extends within the three compartments in a corrugated manner, which can be understood in that the tube 6 has a shape of a wave or a snake within all three compartments when seen in a top view. The corrugation of the tube 6 extends in x- and y-directions of the plane formed by the bottom of the device 1 for preparing a DNA product or the plane formed by the floor. The corrugated form of the tube 6 defines a dwell time of the PCR liquid in the respective compartment. The corrugation direction of the tube 6 is essentially perpendicular to the stacking direction of the tube 6.

[0154] As shown in FIG. 1, the device 1 for preparing a DNA product further comprises a PCR liquid inlet 20 for a PCR mix, which is connectable to an external pump unit or pump (37) for pumping the PCR liquid through the tube 6. The device 1 for preparing a DNA product further comprises a DNA product outlet 25 for a PCR liquid product. Alternatively, the PCR device may comprise an internal pump unit or pump (37) for pumping the PCR liquid through the tube 6. As shown in FIG. 1, the PCR liquid inlet 20 may be located at the first compartment, whereas the DNA product outlet 25 may be located at the third compartment.

[0155] As shown in FIGS. 1 and 2, the device 1 for preparing a DNA product in this embodiment further comprises a thermal medium inlet 12 and a thermal medium outlet 13 in each of the three compartments. As noted above, it may comprise other means to adjust the temperature in each compartment. If thermal medium is used, the thermal medium may enter each of the three compartments by the respective thermal medium inlet 12 and may exit each of the three compartments by the respective thermal medium outlet 13. This may lead to a flow of thermal medium through the respective compartment. The medium outlet 13 is preferably positioned at the top to facilitate the removal of gas bubbles.

[0156] The thermal medium can be understood as a liquid heat carrier and can be used to provide three different temperature zones in the three compartments to prepare the DNA product by thermal cycling. The three temperatures zones correspond to the temperature profiles of the PCR for denaturation, annealing, and elongation. The PCR liquid is separated by the tube walls from the thermal medium in the compartments. The tube walls allow a temperature exchange between the PCR liquid and the inside of the compartments, e.g. the thermal medium, which can be used to heat or cool the PCR liquid, e.g. by means of the thermal medium.

[0157] The thermal medium inlet 12 and the thermal medium outlet 13 are in this embodiment arranged within each compartment to provide the thermal medium flow through the compartment essentially in a counter direction to a guiding direction of the PCR liquid in the tube 6. As shown in FIG. 1, this can be understood that in case of a guiding direction of the PCR liquid from top left to bottom right, the thermal medium inlet 12 and the thermal medium outlet 13 are arranged to provide the thermal medium flow in a counter direction from bottom right to top left (see the arrow in the first compartment 7). The counter flow of thermal medium and PCR liquid improves a temperature exchange between the thermal medium and the PCR liquid.

[0158] As shown in FIG. 8, the device 1 in this embodiment comprises an isolation 28 and a shell 29 as well as an inner-compartment wall 30 that surrounds the tube in a corrugated matter in each of the compartments 7, 8 and 15. Each compartment comprises at least one heating unit 31 (here depicted as heating cartridge) as well as a cooling unit, here comprising a heat pipe 32, which can be used for cooling, if necessary. In this embodiment, the space in each compartment surrounded by the inner-compartment wall 30 (wherein this space surrounds the tube 6) is filled with a thermal solid (such as e.g. aluminum-pellets), optionally in combination with a liquid or gel, wherein the thermal solid is heated by the at least one heating unit 31 as well asif necessarycooled by a cooling unit (comprising heat pipe 32) in order to provide a specific temperature in each compartment. The space between the compartments and/or the surrounding shell 29, i.e. the space that does not surround the tube 6, can be filled with synthetic material (e.g. synthetic balls or granules). FIG. 9 shows the top view of the device 1 shown in FIG. 8, wherein the isolation 28 and the shell 29 are depicted, and also the top of the heating unit 31 as well as the heat pipe 32. The heat pipe 32 is connected to a thermoelectric cooler 33 (e.g. a copper-block with a Peltier element) and a ventilator 34 to transport the heat away. Also in this embodiment, three different temperature zones are provided in the three compartments to prepare the DNA product by thermal cycling. The three temperatures zones correspond to the temperature profiles of the PCR for denaturation, annealing, and elongation. The PCR liquid is separated by the tube walls from the thermal solid in the compartments.

[0159] As shown in FIGS. 1 and 8, the device 1 for preparing a DNA product further comprises septum walls 14 between the compartments to insulate the compartments from each other (in FIG. 8, the septum walls 14 are present only partially as the inner-compartment walls 30 already separate the compartments from each other). One septum wall 14 is arranged between the first compartment 7 and the second camber, another septum wall 14 is arranged between the second camber compartment and the third camber and a further septum wall 14 is arranged between the third camber compartment and the first camber. As a consequence, the adjacent compartments are thermally insulated from each other and therefore different temperatures can be established and maintained in the adjacent compartments. A septum wall 14 can be a plate shaped element. The septum walls 14 between the compartments can be implemented by several walls or by only one (e.g. T-shaped or star shaped) wall. The septum walls 14 comprise through holes for the tube 6 to extend from one compartment to the next.

[0160] As shown in FIGS. 1 and 9, the device 1 for preparing a DNA product further comprises sensors 21, here e.g. temperature sensors 21, arranged in each compartment. It is preferred that several of these temperature sensors are present in each compartment.

[0161] The device 1 for preparing a DNA product further comprises stands 17 for holding the tube 6. FIG. 4 shows a lateral 3D view of a tube 6 wound around several stands 17. The stands 17 are also shown in FIGS. 1 and 8. The stands 17 are arranged in each compartment to hold and guide the tube 6. The stands 17 provide a fixation point as well as a deviation point for the corrugation of the tube 6. The stands 17 may therefore comprise slits or the like to mount the tube 6 (winding) to the stand 17. The tube 6 extends partially around an outer dimension of the stand 17. The stands 17 here have a T-shape or an L-shape, when seen in a top view, but also other shapes are possible, e.g. a cylindrical shape. The stands 17 may be exchangeable and repositionable in the compartment. The tube 6 is here the same throughout the device 1 for preparing a DNA product and may be exchangeable.

[0162] FIG. 5 shows a 3D view of a manufacturing device 10 for a pharmaceutical product. FIG. 11 shows a cut-out thereof. The manufacturing device 10 for a pharmaceutical product comprises a housing 5, a process chamber 2, a technical chamber 3 and a device 1 for preparing a DNA product by means of Capillary Polymerase Chain Reaction as described above. The pharmaceutical product can be understood as an active pharmaceutical ingredient or any precursor or any intermediate thereof (e.g. a DNA vaccine or a DNA template for RNA synthesis). The manufacturing device 10 for a pharmaceutical product can be operated under GMP (guidelines for good manufacturing practice)-compliant conditions.

[0163] The manufacturing device 10 for a pharmaceutical product can be understood as a closed and sealed housing 5 encompassing different chambers for different purposes. The process chamber 2 is suitable and used for manufacturing the pharmaceutical product. The process chamber 2 is here a grade A room. The process chamber 2 is sealed relative to the technical chamber 3. The technical chamber 3 is suitable and used for housing apparatus, as e.g. a pump, a motor, a mixer, a processor or the like. The chambers are separated by a separation element 4, which can be understood as a plate. The separation element 4 extends through the housing 5 and can be used to mount apparatus thereto.

[0164] The technical chamber 3 and the process chamber 2 are arranged so that a gas flow in the technical chamber 3 is parallel to a gas flow in the process chamber 2, but in an opposite direction. The gas of the gas flow is here clean air. There is a gas inlet duct 9a and a gas outlet duct 9b.

[0165] At least one control unit controls the gas flow through the process chamber 2 to provide a gas shower falling in the direction of gravity (downwards) and/or a positive pressure in the process chamber 2. The control unit can be a valve or a processor. The gas shower can be laminar. The gas flow can be provided by a flow unit, which may be arranged within or outside the housing 5 and may communicate with the control unit. The flow unit may be a pump, a HVAC system or the like. The positive pressure is an overpressure relative to the environment outside the housing 5. The flow unit provides variable volumes of gas, while the control unit adjusts different pressures in the gas flow.

[0166] As shown in FIG. 5, the separation element 4 comprises at least an appendix 16 with an opening towards the process chamber 2 and an extension (extended process chamber) in the direction of the technical chamber 3. The appendix 16 is used to house apparatuses, as e.g. the above described device 1 for preparing a DNA product (which may also be referred to as PCR reactor). This allows increasing the available space in the process chamber 2. The device 1 for preparing a DNA product is here housed in a box 26, which is inserted as a unitary element into the appendix 16. The box 26 may be exchangeable.

[0167] As also shown in FIG. 5, a front face of the device 1 for preparing a DNA product and in particular a front face of the box 26 ends essentially flush with the opening of the appendix 16 into the process chamber 2 when the device 1 for preparing a DNA product is inserted into the appendix 16. As a result, the gas flow and in particular the gas shower is seamless and may be not or less disturbed and/or precipitation surfaces are reduced. The appendix 16 and here its front face is closed by a cover. The cover may be part of the box 26 of the device 1 for preparing a DNA product

[0168] As shown in FIG. 5, the manufacturing device 10 for a pharmaceutical product further comprises a fluid collect tray 11 to collect fluid leakages. The fluid collect tray 11 can be arranged at a bottom of the housing 5 and/or in the passage from the process chamber 2 to the technical chamber 3. The fluid collect tray 11 can be equipped with a leakage sensor 21.

[0169] FIG. 11 shows the back of the manufacturing device 10 where the device 1 is inserted. At the back, which faces the technical chamber 3, a ventilator 34 is present. Further, an air inlet 35 is present, along with electronic plugs 36 to connect the device 1 to technical media, in particular power, in the technical chamber. This embodiment relates in particular to devices where a thermal solid is used for adjusting the temperature of the compartments (e.g. as exemplified in FIGS. 8, 9, and 10).

[0170] FIGS. 6a and 6b as well as FIG. 10 show 3D views of the box 26 housing the device 1 for preparing a DNA product to be inserted into the manufacturing device 10 for a pharmaceutical product. The box 26 is closed by the cover. The cover comprises handles 22 for handling the device 1 for preparing a DNA product in and/or out of the appendix 16. The box 26 comprises inlets 12 and outlets 13 for the thermal media for each compartment of the device 1 for preparing a DNA product (see FIG. 6b) or heating cartridges 31 as well as heat pipes 32 connected to a thermoelectric cooler 33 and a ventilator 34 (see FIG. 10). An intake for a PCR liquid 23 and/or an outfall for a DNA product 24 is/are positioned at the front face and here at the cover of the box 26 of the device 1 for preparing a DNA product. The appendix 16 (see FIG. 5) and the box 26 comprise corresponding parts of an exchange rail system 27 to guide and ease an exchange of the apparatus in the appendix 16, as e.g. exchange the device 1 for preparing a DNA product by another device. As shown in FIG. 6b and FIG. 10, a sensor 21, e.g. a leakage sensor 21 or a temperature sensor 21, is arranged in the box 26. The box 26 further comprises connections to be connected to the process chamber 2, e.g. electric connections. In the embodiment according to FIG. 10, the sensor is preferably a temperature sensor as the embodiment of FIG. 10 (where a thermal solid is used as heating means) typically does not comprise a leakage sensor (as no thermal medium is used in the embodiment shown in FIG. 10).

[0171] An external pump unit or pump (37 in FIG. 7) for pumping the PCR liquid through the tube 6 of the device 1 for preparing a DNA product may be arranged outside the device 1, and may e.g. be located in the process chamber of the manufacturing device 10 for a pharmaceutical product.

[0172] FIG. 7 shows schematically and exemplarily a manufacturing module 100 for a pharmaceutical product according to the invention. The manufacturing module 100 comprises here four manufacturing devices 10. The manufacturing devices are very flexible and several manufacturing devices can be adapted to several manufacturing steps to form e.g. a manufacturing chain.

[0173] The four manufacturing devices 10 are coupled with each other by means of a coupling unit 18 arranged at an outer wall of the housing 5. The coupling unit 18 allows a fluid communication and coupling, and preferably also a mechanical coupling as well as a data communication and coupling of the manufacturing devices 10. The four manufacturing devices 10 are exemplary shown herein to be (from the left) a first device comprising a process media supply unit and a mixing unit, a second device comprising a manufacturing device 10 for a pharmaceutical product as described above (a device 1 for preparing a DNA product by means of Capillary Polymerase Chain Reaction), a third device comprising a purification unit and a fourth device comprising a filtration unit, wherein the filtration unit comprises two filtration units, namely a tangential flow filtration unit and a sterile filter. This exemplary module can be used for the production of DNA and the amplification as well as purification of DNA, respectively. The exemplary module used for the production/amplification of DNA may be connected to further manufacturing devices that transcribe the obtained DNA into RNA (e.g. in a manufacturing device comprising an RNA bioreactor as described in WO2020002598).

[0174] The manufacturing module 100 comprises an attachment element 19 arranged at an outer wall of the housing 5 of the manufacturing device 10 outside the housing 5. The attachment element 19 is configured to hold a media supply (not shown) e.g. in form of a media reservoir.

Definitions

[0175] For the sake of clarity and readability the following definitions are provided. Any technical feature mentioned for these definitions may be read on each and every embodiment of the invention. Additional definitions and explanations may be specifically provided in the context of these embodiments.

[0176] As used in the specification and the claims, the singular forms of a and an also include the corresponding plurals unless the context clearly dictates otherwise.

[0177] The term about in the context of the present invention denotes an interval of accuracy that a person skilled in the art will understand to still ensure the technical effect of the feature in question. The term typically indicates a deviation from the indicated numerical value of 10% and preferably 5%.

[0178] It needs to be understood that the term comprising is not limiting. For the purposes of the present invention, the term consisting of is considered to be a preferred embodiment of the term comprising of. If hereinafter a group is defined to comprise at least a certain number of embodiments, this is also meant to encompass a group which preferably consists of these embodiments only.

[0179] The term pharmaceutical product as used herein relates to an active pharmaceutical ingredient or any precursor or any intermediate thereof. Thus, a pharmaceutical product may in particular be the active pharmaceutical ingredient that is used in a medicament and administered to a human or animal subject in order to treat or prevent a disease, i.e. it has a clinical grade, particularly when it comes to parameters such as purity, integrity. Such products are typically produced in vitro in a synthetic process, and the process of production includes precursors and intermediates. For the present invention, it is particularly preferred that the active pharmaceutical ingredient is a biomolecule, in particular a nucleic acid. The nucleic acid may be DNA. The DNA may particularly be used as a vaccine or as a DNA template for RNA in vitro transcription. When producing e.g. an mRNA as the active pharmaceutical ingredient, a first step may be the production of template DNA (also referred to as a DNA template herein), wherein this template DNA corresponds to a precursor of the mRNA since it serves as template in the in vitro transcription reaction when producing the RNA. An intermediate of the DNA production may be the DNA that is obtained from the PCR reaction before purification since such a DNA does not correspond to the final active pharmaceutical ingredient but inter alia requires that purification steps are carried out in order to provide the product in the required clinical grade.

[0180] The term DNA is the usual abbreviation for deoxyribonucleic acid. It is a nucleic acid molecule, i.e. a polymer consisting of nucleotide monomers. These nucleotides are usually deoxy-adenosine-monophosphate, deoxy-thymidine-monophosphate, deoxy-guanosine-monophosphate and deoxy-cytidine-monophosphate monomers or analogs thereof which areby themselvescomposed of a sugar moiety (deoxyribose), a base moiety and a phosphate moiety, and polymerize by a characteristic backbone structure. The backbone structure is, typically, formed by phosphodiester bonds between the sugar moiety of the nucleotide, i.e. deoxyribose, of a first and a phosphate moiety of a second, adjacent monomer. The specific order of the monomers, i.e. the order of the bases linked to the sugar/phosphate-backbone, is called the DNA-sequence. DNA may be single stranded or double stranded. In the double stranded form, the nucleotides of the first strand typically hybridize with the nucleotides of the second strand, e.g. by A/T-base-pairing and G/C-base-pairing.

[0181] The term RNA is the usual abbreviation for ribonucleic acid. It is a nucleic acid molecule, i.e. a polymer consisting of nucleotide monomers. These nucleotides are usually adenosine-monophosphate (AMP), uridine-monophosphate (UMP), guanosine-monophosphate (GMP) and cytidine-monophosphate (CMP) monomers or analogs thereof, which are connected to each other along a so-called backbone. The backbone is formed by phosphodiester bonds between the sugar, i.e. ribose, of a first and a phosphate moiety of a second, adjacent monomer. The specific order of the monomers, i.e. the order of the bases linked to the sugar/phosphate-backbone, is called the RNA sequence. RNA can be obtained by transcription of a DNA sequence, e.g., inside a cell. In eukaryotic cells, transcription is typically performed inside the nucleus or the mitochondria. In vivo, transcription of DNA usually results in the so-called premature RNA which has to be processed into so-called messenger-RNA, usually abbreviated as mRNA. Processing of the premature RNA, e.g. in eukaryotic organisms, comprises a variety of different posttranscriptional modifications such as splicing, 5-capping, polyadenylation, export from the nucleus or the mitochondria and the like. The sum of these processes is also called maturation of RNA. The mature messenger RNA usually provides the nucleotide sequence that may be translated into an amino acid sequence of a particular peptide or protein. Typically, a mature mRNA comprises a 5-cap, optionally a 5UTR, a coding sequence, optionally a 3UTR and a poly(A) sequence. If RNA molecules are of synthetic origin, as in the present invention, the RNA molecules are meant not to be produced in vivo, i.e. inside a cell or purified from a cell, but in an in vitro method. An examples for a suitable in vitro method is in vitro transcription. In addition to messenger RNA, several non-coding types of RNA exist which may be involved in regulation of transcription and/or translation, and immunostimulation and which may also be produced by in vitro transcription. The term RNA further encompasses RNA molecules, such as viral RNA, retroviral RNA and replicon RNA, small interfering RNA (siRNA), antisense RNA, saRNA (small activating RNA), CRISPR RNA (small guide RNA, sgRNA), ribozymes, aptamers, riboswitches, immunostimulating RNA, transfer RNA (tRNA), ribosomal RNA (rRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), microRNA (miRNA), and Piwi-interacting RNA (piRNA).

[0182] The term RNA in vitro transcription relates to a process wherein RNA is synthesized in a cell-free system. RNA may be obtained by DNA-dependent RNA in vitro transcription of an appropriate DNA template, which may be a linearized plasmid DNA template or a PCR-amplified DNA template. The promoter for controlling RNA in vitro transcription can be any promoter for any DNA-dependent RNA polymerase. Particular examples of DNA-dependent RNA polymerases are the T7, T3, SP6, or Syn5 RNA polymerases.

[0183] Reagents used in RNA in vitro transcription typically include: a DNA template (linearized DNA or linear PCR product) with a promoter sequence that has a high binding affinity for its respective RNA polymerase such as bacteriophage-encoded RNA polymerases (T7, T3, SP6, or Syn5); ribonucleotide triphosphates (NTPs) for the four bases (adenine, cytosine, guanine and uracil); optionally, a cap analogue (e.g. m7G(5)ppp(5)G (m7G) or a cap analogue derivable from the structure disclosed in claim 1-5 of WO2017/053297 or any cap structures derivable from the structure defined in claim 1 or claim 21 of WO2018075827); optionally, further modified nucleotides as defined herein; a DNA-dependent RNA polymerase capable of binding to the promoter sequence within the DNA template (e.g. T7, T3, SP6, or Syn5 RNA polymerase); optionally, a ribonuclease (RNase) inhibitor to inactivate any potentially contaminating RNase; optionally, a pyrophosphatase to degrade pyrophosphate (inhibitor of RNA synthesis); MgCl2, which supplies Mg2+ ions as a co-factor for the polymerase; a buffer (TRIS or HEPES) to maintain a suitable pH value, which can also contain antioxidants (e.g. DTT), and/or polyamines such as spermidine at optimal concentrations, e.g. a buffer system comprising Citrate and/or betaine as disclosed in WO2017/109161.

[0184] The nucleotide mixture used in RNA in vitro transcription may additionally contain modified nucleotides as defined herein. In that context, preferred modified nucleotides comprise pseudouridine (), N1-methylpseudouridine (m1), 5-methylcytosine, and 5-methoxyuridine. The nucleotide mixture (i.e. the fraction of each nucleotide in the mixture) used for RNA in vitro transcription reactions may be optimized for the given RNA sequence, preferably as described in WO2015188933.

[0185] An RNA in vitro transcription (IVT) master mix may comprise the components necessary for performing an RNA in vitro transcription reaction as defined above. Accordingly, an IVT master mix may comprise at least one of the components selected from a nucleotide mixture, a cap analogue, a DNA-dependent RNA polymerase, an RNAse inhibitor, a pyrophosphatase, MgCl2, a buffer, an antioxidant, betaine, Citrate.

[0186] As used herein, the term template DNA (or a DNA template) typically relates to a DNA molecule comprising a nucleic acid sequence encoding the RNA sequence to be transcribed in vitro. The template DNA is used as template for RNA in vitro transcription in order to produce the RNA encoded by the template DNA. Therefore, the template DNA comprises all elements necessary for RNA in vitro transcription, particularly a promoter element for binding of a DNA dependent RNA polymerase as e.g. T3, T7 and SP6 RNA polymerases 5 of the DNA sequence encoding the target RNA sequence. Furthermore, the template DNA may comprise primer binding sites 5 and/or 3 of the DNA sequence encoding the target RNA sequence to determine the presence of the DNA sequence encoding the target RNA sequence e.g. by PCR or DNA sequencing.

[0187] The polymerase chain reaction or PCR is a technology in molecular biology used to amplify a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. The method relies on thermal cycling, consisting of cycles of repeated heating and cooling of the reaction for DNA melting and enzymatic replication of the DNA. Primers (short DNA fragments) containing sequences complementary to the target sequence along with a heat-stable DNA polymerase, such as Taq polymerase, enable selective and repeated amplification. As PCR progresses, the DNA generated is itself used as a template for replication, setting in motion a chain reaction in which the DNA template is exponentially amplified. The DNA polymerase enzymatically assembles a new DNA strand from DNA building-blocks, the nucleotides, by using single-stranded DNA as a PCR template and DNA oligonucleotides (also called DNA primers), which are required for initiation of DNA synthesis. The vast majority of PCR methods use thermal cycling, i.e., alternately heating and cooling the PCR sample through a defined series of temperature steps. In the first step, the two strands of the DNA double helix are physically separated at a high temperature in a process called DNA melting. In the second step, the temperature is lowered and the two DNA strands become templates for DNA polymerase to selectively amplify the target DNA. The selectivity of PCR results from the use of primers that are complementary to the DNA region targeted for amplification under specific thermal cycling conditions.

[0188] A PCR master mix or PCR liquid may comprise the components necessary for performing a PCR as defined above. Accordingly, a PCR master mix may comprise at least one of the components selected from a nucleotide mixture, a DNA polymerase, the synthetic DNA as (initial) template and a buffer.

[0189] The term lipid nanoparticle or LNP refers to a formulation of the pharmaceutical product, in particular the nucleic acid. In the context of the present invention, the term LNP is not restricted to any particular morphology, and includes any morphology generated when a cationic lipid and optionally one or more further lipids are combined, e.g. in an aqueous environment and/or in the presence of an nucleic acid. For example, a liposome, a lipid complex, a lipoplex and the like are within the scope of a lipid nanoparticle (LNP). LNPs typically comprise a cationic lipid and one or more excipients selected from neutral lipids, charged lipids, steroids and polymer conjugated lipids (e.g., PEGylated lipid). The nucleic acid may be encapsulated in the lipid portion of the LNP or an aqueous space enveloped by some or the entire lipid portion of the LNP. In one embodiment, the LNP consists essentially of (i) at least one cationic lipid; (ii) a neutral lipid; (iii) a sterol, e.g. cholesterol; and (iv) a PEG-lipid, e.g. PEG-DMG or PEG-cDMA, in a molar ratio of about 20-60% cationic lipid:5-25% neutral lipid:25-55% sterol; 0.5-15% PEG-lipid.

[0190] The term process medium refers to a component that is directly and physically involved in any reaction or any process step required to produce the pharmaceutical product. Accordingly, as the production is carried out in the process chamber, a process medium will be present in the process chamber when the device is used for production. A process medium can thus be in particular any starting material (such as e.g. a nucleotide), any catalyzing material (such as e.g. an enzyme) and any buffer (such as e.g. a reaction buffer or a purification buffer). A process medium is typically provided in a process medium supply container.

[0191] The term technical medium refers to a component that is not directly involved in any reaction or any process step required to produce the pharmaceutical product. Rather, a technical medium is indirectly involved, e.g. as a power cable that provides power to a unit or apparatus that processes the process medium, and will be located in the technical chamber. A technical medium supply will e.g. be power or power supply provided by a power cable.

[0192] It has to be noted that embodiments of the invention are described with reference to different subject matters. In particular, some embodiments are described with reference to method type claims whereas other embodiments are described with reference to the device type claims. However, a person skilled in the art will gather from the description that, unless otherwise notified, in addition to any combination of features belonging to one type of subject matter also any combination between features relating to different subject matters is considered to be disclosed with this application. However, all features can be combined providing synergetic effects that are more than the simple summation of the features.

[0193] If a group is defined to comprise at least a certain number of embodiments, this is also meant to encompass a group, which preferably consists of these embodiments only.

[0194] While the invention has been illustrated and described in detail in the drawings and the description, such illustration and description are to be considered illustrative or exemplary and not restrictive. The invention is not limited to the disclosed embodiments. Other variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing a claimed invention, from a study of the drawings, the disclosure, and the dependent claims.

[0195] In the claims, the word comprising does not exclude other elements or steps, and the indefinite article a or an does not exclude a plurality. The mere fact that certain measures are re-cited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope.

LIST OF REFERENCES SIGNS

[0196] 1 device for preparing a DNA product [0197] 2 process chamber [0198] 3 technical chamber [0199] 4 separation element [0200] 5 housing [0201] 6 tube [0202] 7 first compartment [0203] 8 second compartment [0204] 9 duct [0205] 9a gas inlet duct [0206] 9b gas outlet duct [0207] 10 manufacturing device [0208] 11 collect tray [0209] 12 thermal medium inlet [0210] 13 thermal medium outlet [0211] 14 septum walls [0212] 15 third compartment [0213] 16 appendix [0214] 17 stand [0215] 18 coupling unit [0216] 19 attachment element [0217] 20 PCR liquid inlet [0218] 21 sensor [0219] 22 handle [0220] 23 intake for PCR liquid [0221] 24 outfall for a DNA product [0222] 25 DNA product outlet [0223] 26 box [0224] 27 exchange rails [0225] 28 isolation [0226] 29 shell [0227] 30 inner-compartment wall [0228] 31 heating unit [0229] 32 heat pipe (part of a cooling unit) [0230] 33 thermoelectric cooler (part of a cooling unit) [0231] 34 ventilator [0232] 35 air inlet [0233] 36 electric plug [0234] 37 pump unit for pumping the PCR liquid [0235] 100 manufacturing module

[0236] A set of embodiments of the present application relates to:

[0237] 1. A device (1) for preparing a DNA product by means of Capillary Polymerase Chain Reaction (PCR), comprising: [0238] a tube (6) for guiding a PCR liquid, [0239] a first compartment (7), and [0240] at least a second compartment (8),
wherein the tube (6) is wound at least from the first compartment (7) to the second compartment (8), and
wherein the first compartment (7) and the second compartment (8) each comprise a thermal medium inlet (12) and a thermal medium outlet (13) to provide a separate thermal medium flow through the first compartment (7) and the second compartment (8) to adjust a separate temperature in the first compartment (7) and the second compartment (8) to prepare the DNA product based on the PCR liquid.

[0241] 2. Device (1) for preparing a DNA product according to embodiment 1, wherein septum walls (14) are arranged between the compartments to insulate the compartments from each other.

[0242] 3. Device (1) for preparing a DNA product according to the preceding embodiment, wherein the septum walls (14) comprise through holes for the tube (6) to extend from one compartment to the next.

[0243] 4. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the thermal medium inlet (12) and the thermal medium outlet (13) are arranged within at least one and preferably within each compartment to provide the thermal medium flow through the compartment essentially in a counter direction to a guiding direction of the PCR liquid in the tube (6).

[0244] 5. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the thermal medium inlet (12) is arranged at a lower position in each of the compartments and the thermal medium outlet (13) is arranged at an upper position in each of the compartments.

[0245] 6. Device (1) for preparing a DNA product according to one of the preceding embodiments, further comprising a third compartment (15), wherein the tube (6) is wound at least from the first compartment (7) to the second compartment (8) and from the second compartment (8) to the third compartment (15).

[0246] 7. Device (1) for preparing a DNA product according to the preceding embodiment, wherein the tube (6) is further wound from the third compartment (15) to the first compartment (7), from the first compartment (7) to the second compartment (8), and from the second compartment (8) to the third compartment (15).

[0247] 8. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) wound from compartment to compartment forms a helical stack of tube windings.

[0248] 9. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) extends within at least one and preferably in each of the compartments in a corrugated manner.

[0249] 10. Device (1) for preparing a DNA product according to embodiments 8 and 9, wherein a stacking direction of the helical stack of tube windings is different and preferably essentially perpendicular to a corrugation direction of the corrugated tube (6).

[0250] 11. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein each compartment forms a basin to be filled by the thermal medium.

[0251] 12. Device (1) for preparing a DNA product according to one of the preceding embodiments, further comprising at least a stand (17) arranged in one of the compartments to provide a fixation point for the tube (6) and/or a deviation point for a corrugation of the tube (6).

[0252] 13. Device (1) for preparing a DNA product according to the preceding embodiment, wherein the tube (6) extends partially or fully or more than once around an outer dimension of the stand (17).

[0253] 14. Device (1) for preparing a DNA product according to one of the embodiment 12 to 13, wherein the stand (17) is exchangeable and/or repositionable in the compartment.

[0254] 15. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) comprises a PCR liquid inlet (20), which is connectable to a pump unit for pumping the PCR liquid through the tube (6).

[0255] 16. Device (1) for preparing a DNA product according to the preceding embodiment, wherein the pump unit is arranged outside the compartments and preferably outside a shell (5) surrounding the compartments.

[0256] 17. Device (1) for preparing a DNA product according to embodiment 15 or 16, wherein the pump unit is configured to provide a flow rate of the PCR liquid in a range of about 0.1 to about 10 mL/min, preferably about 0.2 to about 3 mL/min.

[0257] 18. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) comprises a DNA product outlet (25) for a continuous preparation of the DNA product.

[0258] 19. Device (1) for preparing a DNA product according to embodiment 1 to 17, wherein the tube (6) comprises a closable DNA product outlet (25) for a discontinuous preparation of the DNA product.

[0259] 20. Device (1) for preparing a DNA product according to one of the preceding embodiments, further comprising at least a sensor (21) arranged in at least one of compartments, wherein the sensor (21) is configured to detect a temperature, a flow rate or a leakage of the compartment.

[0260] 21. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) has a diameter in a range of about 0.5 to about 1.5 mm, preferably about 0.75 to about 1.25 mm.

[0261] 22. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) has a length between a tube inlet and a tube outlet in a range of about 10 to about 200 m, preferably about 25 to about 50 m.

[0262] 23. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) is dimensioned to contain PCR liquid in a range of about 10 mL to about 1 L, preferably about 20 mL to about 250 mL.

[0263] 24. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the helical stack of tube windings comprises about 10 to about 50 windings, preferably about 15 to about 40.

[0264] 25. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the device (1) for preparing a DNA product is configured to prepare about 1 mg and more of the DNA product per reaction, preferably in a range of about 1 to about 30 mg, more preferably in a range of about 15 to about 25 mg.

[0265] 26. Device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the device (1) for preparing a DNA product is configured to prepare the DNA product in a range of about 50 g to about 150 g per mL PCR liquid, preferably about 75 to about 125 g per mL PCR liquid.

[0266] 27. A manufacturing device (10) for a pharmaceutical product, comprising a process chamber (2), a technical chamber (3), and a device (1) for preparing a DNA product by means of Capillary Polymerase Chain Reaction according to one of the preceding embodiments.

[0267] 28. Manufacturing device (10) for a pharmaceutical product according to the preceding embodiment, further comprising a control unit configured to control a gas flow through the process chamber (2) as a gas shower.

[0268] 29. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 28, wherein at least the process chamber (2), preferably the manufacturing device (10), is operable according to the requirements of the EU guidelines for good manufacturing practice (GMP) for medicinal products, annex 1.

[0269] 30. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 29, further comprising a separation element (4) separating the process chamber (2) from the technical chamber (3), wherein the separation element (4) comprises an appendix (16) with an opening towards the process chamber (2) and protruding in the direction of the technical chamber (3), and wherein the device (1) for preparing a DNA product is insertable into the appendix (16).

[0270] 31. Manufacturing device (10) for a pharmaceutical product according to the preceding embodiment, wherein a front face of the device (1) for preparing a DNA product ends essentially flush with the opening of the appendix (16) into the process chamber (2) when the device (1) for preparing a DNA product is inserted into the appendix (16).

[0271] 32. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 31, wherein at least a handle (22) for handling the device (1) for preparing a DNA product in and/or out of the appendix (16) is positioned at the front face of the device (1) for preparing a DNA product.

[0272] 33. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 32, wherein an intake for a PCR liquid (23) and/or an outfall for a DNA product (24) is/are positioned at the front face of the device (1) for preparing a DNA product.

[0273] 34. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 33, wherein an intake for a thermal medium and/or an outfall for the thermal medium is/are positioned at a back plate of the device (1) for preparing a DNA product opposite to the front face of the device (1) for preparing a DNA product.

[0274] 35. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 34, further comprising a leakage sensor (21) arranged in the appendix (16) outside the device (1) for preparing a DNA product.

[0275] 36. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 35, wherein the appendix (16) comprises exchange rails (27) to guide an exchange of the device (1) for preparing a DNA product with another device (1) for preparing a DNA product.

[0276] 37. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 36, wherein a pump unit for pumping the PCR liquid through a tube (6) of the device (1) for preparing a DNA product is arranged outside a casing of the manufacturing device (10) for a pharmaceutical product.

[0277] 38. Manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 37, further comprising a coupling unit (18) arranged at the casing of the manufacturing device (10) for a pharmaceutical product and configured to couple the manufacturing device (10) for a pharmaceutical product to another manufacturing device (10).

[0278] 39. A manufacturing module (100) for a pharmaceutical product, comprising a manufacturing device (10) for a pharmaceutical product according to one of the embodiments 27 to 38 and at least one of a group comprising a manufacturing device (10) with a process media supply unit and a mixing unit, a manufacturing device (10) with a purification unit, and a manufacturing device (10) with a filtration unit.

[0279] 40. A method for preparing a DNA product by means of Capillary Polymerase Chain Reaction (PCR), comprising: [0280] providing a first compartment (7), a second compartment (8), and a tube (6), wherein the tube (6) is wound at least from the first compartment (7) to the second compartment (8), [0281] providing a separate thermal medium flow through each compartment by means of a thermal medium inlet (12) and a thermal medium outlet (13) in each compartment to adjust a separate temperature in each compartment, and [0282] guiding a PCR liquid through the tube (6) and thereby through the separate temperatures in each compartment to prepare the DNA product based on the PCR liquid.

[0283] 41. Method according to the preceding embodiment, wherein a first temperature in the first compartment (7) is in a range of about 85 C. to about 105 C., preferably about 98 C.

[0284] 42. Method according to one of the embodiments 40 to 41, wherein a second temperature in the second compartment (8) is in a range of about 45 C. to about 72 C., preferably about 60 C. to about 72 C.

[0285] 43. Method according to one of the embodiments 40 to 42, further comprising providing a third compartment (15), wherein the tube (6) is wound at least from the second compartment (8) to the third compartment (15), and wherein a third temperature in the third compartment (15) is in a range of about 65 C. to about 75 C., preferably about 72 C.

[0286] 44. Method according to one of the embodiments 40 to 43, wherein the first temperature in the first compartment (7) is configured for a denaturation step of the Polymerase Chain Reaction, wherein the second temperature in the second compartment (8) is configured for an annealing step of the Polymerase Chain Reaction and optionally also for an elongation step of the Polymerase Chain Reaction.

[0287] 45. Method according to embodiment 43, wherein the third temperature in the third compartment (15) is configured for an elongation step of the Polymerase Chain Reaction.

[0288] 46. Use of a device according to one of the embodiments 1 to 26 for preparing a DNA product by means of Capillary Polymerase Chain Reaction.

[0289] 47. Use of a manufacturing device (10) according to one of the embodiments 27 to 38 for a production of a pharmaceutical product.

[0290] A set of preferred embodiments of the present application relates to:

[0291] 1. A device (1) for amplifying DNA by Polymerase Chain Reaction (PCR), comprising: [0292] a tube (6) for guiding a PCR liquid, [0293] a first compartment (7), [0294] a second compartment (8), and [0295] a third compartment (15),
wherein the tube (6) is wound through the first compartment (7) and from the first compartment (7) to and through the second compartment (8), from the second department (8) to and through the third compartment (15), and from the third compartment (15) to the first compartment (7), wherein the tube (6) wound from compartment to compartment forms a helical stack of tube windings and each tube winding represents one PCR cycle, wherein the tube (6) comprises a PCR liquid inlet (20) and a DNA product outlet (25), and
wherein the first compartment (7), the second compartment (8) and the third compartment (15) each comprises means to adjust the temperature in the compartment to prepare the DNA product based on the PCR liquid, wherein the first compartment is configured to provide a temperature for denaturation, the second compartment (8) is configured to provide a temperature for annealing and the third compartment (15) is configured to provide a temperature for elongation.

[0296] 2. The device (1) for amplifying DNA according to embodiment 1, wherein the PCR liquid inlet (20) is connectable to a pump unit for pumping the PCR liquid through the tube (6).

[0297] 3. The device (1) for amplifying DNA according to embodiment 1 or 2, wherein septum walls (14) are arranged between the compartments to insulate the compartments from each other, wherein the septum walls (14) comprise through holes for the tube (6) to extend from one compartment to the next.

[0298] 4. The device (1) for amplifying DNA according to any one of embodiments 1 to 3, wherein each compartment comprises a heating unit (31) and optionally a cooling unit (32, 33) as the means to adjust the temperature in each compartment.

[0299] 5. The device (1) for amplifying DNA according to any one of embodiments 1 to 3, wherein each compartment comprises a thermal medium inlet (12) and a thermal medium outlet (13) as the means to adjust the temperature in each compartment, namely to provide a separate thermal medium flow through the first compartment (7), the second compartment (8) and the third compartment (15) to adjust a separate temperature in the first compartment (7), the second compartment (8) and the third compartment (15).

[0300] 6. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the tube (6) extends within each of the compartments in a corrugated manner.

[0301] 7. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein each compartment forms a basin to be filled with a thermal medium or a thermal solid.

[0302] 8. The device (1) for amplifying DNA according to any one of the preceding embodiments, further comprising at least a stand (17) arranged in one of the compartments to provide a fixation point for the tube (6) and/or a deviation point for a corrugation of the tube (6).

[0303] 9. The device (1) for amplifying DNA according to embodiment 8, wherein the stand (17) is exchangeable and/or repositionable in the compartment.

[0304] 10. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the tube (6) has a diameter in a range of about 0.5 mm to about 50 mm, preferably about 0.5 mm to about 1.5 mm, more preferably about 0.75 to about 1.25 mm.

[0305] 11. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the tube (6) has a length between a tube inlet and a tube outlet in a range of about 10 m to about 200 m, preferably about 50 m to about 100 m.

[0306] 12. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the tube (6) is dimensioned to contain PCR liquid in a range of about 10 mL to about 500 mL, preferably about 20 mL to about 100 mL.

[0307] 13. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the helical stack of tube windings comprises about 10 to about 50 windings, preferably about 15 to about 40.

[0308] 14. A manufacturing device (10) comprising a device (1) for amplifying DNA according to any one of embodiments 1 to 13.

[0309] 15. The manufacturing device (10) according to embodiment 14, further comprising a process chamber (2) and a technical chamber (3).

[0310] 16. The manufacturing device (10) according to embodiment 15, further comprising a control unit configured to control a gas flow through the process chamber (2) as a gas shower.

[0311] 17. The manufacturing device (10) according to embodiment 15 or 16, further comprising a separation element (4) separating the process chamber (2) from the technical chamber (3), wherein the separation element (4) comprises an appendix (16) with an opening towards the process chamber (2) and protruding in the direction of the technical chamber (3), and wherein the device (1) for amplifying DNA is insertable into the appendix (16).

[0312] 18. The manufacturing device (10) according to embodiment 17, wherein a front face of the device (1) for preparing a DNA product ends essentially flush with the opening of the appendix (16) into the process chamber (2) when the device (1) for preparing a DNA product is inserted into the appendix (16).

[0313] 19. A manufacturing module (100), comprising a manufacturing device (10) according to any one of embodiments 14 to 18 and at least one manufacturing device selected from the group consisting of a manufacturing device (10) comprising a process media supply unit and a mixing unit, a manufacturing device (10) comprising a purification unit, and a manufacturing device (10) comprising a filtration unit.

[0314] 20. A method for amplifying DNA by means of Polymerase Chain Reaction (PCR), comprising: [0315] providing a first compartment (7), a second compartment (8), a third compartment (15) and a tube (6), wherein the tube (6) is wound through the first compartment (7) and from the first compartment (7) to and through the second compartment (8) and from the second compartment (8) to and through the third compartment (15), and from the third compartment (15) to the first compartment (7), wherein the tube (6) wound from compartment to compartment forms a helical stack of tube windings, [0316] providing means to adjust a separate temperature in each compartment, and [0317] guiding a PCR liquid through the tube (6) and thereby through the separate temperatures in each compartment to prepare the DNA product based on the PCR liquid.

[0318] 21. The method according to embodiment 20, wherein the temperature in the first compartment (7) is for denaturation, preferably in the range of about 85 C. to about 105 C., preferably about 98 C.

[0319] 22. The method according to embodiment 20 or 21, wherein the temperature in the second compartment (8) is for annealing, preferably in the range of about 45 C. to about 72 C., preferably about 60 C. to about 72 C.

[0320] 23. The method according to any one of embodiments 20 to 22, wherein the temperature in the third compartment (15) is for elongation, preferably in the range of about 65 C. to about 75 C., preferably about 72 C.

[0321] 24. Use of a device (1) according to any one of embodiments 1 to 13 for preparing a DNA product by means of Polymerase Chain Reaction.

[0322] 25. Use of a manufacturing device (10) according to any one of embodiments 14 to 18 or of a manufacturing module (100) according to embodiment 19 for the production of a pharmaceutical product.