Device for preparing DNA products by capillary polymerase chain reaction

Abstract

The present invention relates to a device for preparing a DNA product by capillary polymerase chain reaction (PCR), a device for manufacturing a pharmaceutical product, a module for manufacturing a pharmaceutical product, a method for preparing a DNA product by capillary polymerase chain reaction, the use of the device for preparing a DNA product by polymerase chain reaction, and the use of the device for the production of pharmaceutical products. The device for preparing a DNA product by capillary polymerase chain reaction comprises a tube for conducting a PCR liquid, a first compartment, and at least a 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, the tube wound from compartment to compartment forming a helical stack of tube windings. The first compartment and the second compartment each comprise a means for adjusting the temperature in the compartment to prepare a DNA product based on the PCR liquid, the first compartment being configured to provide a temperature for denaturation and the second compartment being configured to provide a temperature for annealing and / or extension.

Description

[Technical field]

[0001] The present invention relates to a device for preparing a DNA product by capillary polymerase chain reaction, a pharmaceutical production device, a pharmaceutical production module, a method for preparing a DNA product by capillary polymerase chain reaction, the use of the device for preparing a DNA product by polymerase chain reaction and the use of the device for the production of pharmaceutical products. [Background technology]

[0002] Active pharmaceutical ingredients (APIs) in the form of biomolecules are used today on a large scale in the treatment of various diseases. Such biomolecules are, for example, therapeutic antibodies, peptides, and nucleic acids. Therapeutic nucleic acids include DNA and RNA molecules.

[0003] RNA represents a new class of drugs. RNA-based therapeutics include mRNA molecules that code for antigens for use as vaccines. For the production of RNA, typically a DNA template is used in an RNA in vitro transcription reaction. Such a DNA template can be obtained by polymerase chain reaction (PCR). Thus, the production of DNA in the sense of amplification of DNA by PCR is a key step for the production of RNA as an API.

[0004] Additionally, DNA itself may function as the API, for example a DNA vaccine against coronavirus is currently in clinical development. Polymerase chain reaction (PCR) can be used to produce DNA in large quantities, i.e. to amplify the DNA.

[0005] Currently established manufacturing processes for APIs in the form of nucleic acids approved by regulatory agencies typically involve many separate manufacturing steps, all of which must comply with GMP standards. Such steps are typically performed in GMP-compliant rooms, which are not portable since they are located within the production plant. Furthermore, these rooms are typically set up for the production of a specific biomolecule, and therefore are inflexible and must be rebuilt to a fairly large extent if another nucleic acid needs to be produced. In summary, the production of API-grade nucleic acids requires extensive manual work in fixed, inflexible, GMP-regulated laboratories performed by well-trained technicians. As a result, currently established manufacturing processes are time-consuming, cost-intensive, and require a lot of laboratory space and laboratory equipment. Summary of the Invention [Problem to be solved by the invention]

[0006] As outlined above, the common manufacturing process of nucleic acids, specifically DNA and RNA, as APIs or as intermediates in the manufacturing process of APIs, suffers from the challenge that the process is rather inflexible, not portable, and typically requires a significant degree of manual work by well-trained technicians. There is therefore a need to provide improved devices, specifically for preparing / amplifying DNA, and corresponding more flexible manufacturing devices comprising such devices. The manufacturing devices, specifically, should offer the advantage of being operated under GMP-compliant conditions, portable, and / or automated. Such manufacturing devices significantly reduce the time to produce APIs (e.g., DNA) or intermediates (e.g., DNA that can serve as a template for RNA production, where the RNA is the API), which is particularly important in situations of virus outbreaks (e.g., pandemic or local outbreaks). Such portable API / intermediate manufacturing devices, specifically comprising a device for preparing / amplifying DNA, which may be referred to as a "PCT reactor," can be easily transported, installed, and operated in pandemic hotspot regions around the world, thus enabling rapid production of vaccines in those regions. It is therefore of utmost importance that the manufacturing device of the present invention is configured to produce nucleic acid API / intermediates under GMP compliant conditions and that the corresponding "PCR reactor" is configured to be operable in such a manufacturing device.

[0007] The above problems are solved by the subject matter of the independent claims, further embodiments of which are provided in the dependent claims. [Means for solving the problem]

[0008] In a first aspect, the present invention is directed to a device for preparing a DNA product by capillary polymerase chain reaction (PCR). "Preparing a DNA product by capillary PCR" can alternatively be referred to as "amplifying DNA by capillary PCR" or "amplifying DNA by PCR". The device comprises a tube for conducting a PCR solution, a first compartment and at least a second compartment. The tube is wound at least from the first compartment to the second compartment and from the second compartment back to the first compartment, the tube wound from compartment to compartment forming a helical stack of tube turns. Typically, each tube turn represents one PCR cycle. The first compartment and the second compartment each comprise a means for adjusting the temperature in the compartment to prepare a DNA product based on the PCR solution, the first compartment being configured to provide a temperature for denaturation and the second compartment being configured to provide a temperature for annealing and / or extension.

[0009] In a preferred embodiment, the device for preparing DNA products by capillary polymerase chain reaction (PCR) is not a microfluidic device or is not dimensioned to operate as a microfluidic device. In a preferred embodiment, the device for preparing DNA products by capillary polymerase chain reaction (PCR) is not dimensioned or configured to operate as an analytical PCR device. In a preferred embodiment, the device for preparing DNA products by capillary polymerase chain reaction (PCR) is not dimensioned or configured to operate as a device for preparative production of DNA (e.g., for the production of more than 1 mg of DNA product, optionally for the production of g quantities of DNA, e.g., for the production of 5 g of DNA). When operated in a continuous mode, the output is essentially unlimited, so that very large amounts of DNA, e.g., g quantities, can be provided, particularly DNA that serves as template DNA in a subsequent RNA in vitro transcription reaction, when the overall purpose is to produce RNA.

[0010] In one embodiment, the device is not constructed to be capable of recirculating PCR fluids within the device and / or within the tubes contained in the device, but rather, as described above, the tubes are wound into a helical stack of tube windings, and the tubes are not connected to the previous section of the tube at any point (thus, the PCR fluids enter the tube at a first end of the tube, remain in the tube during all PCR cycles, and exit the tube at a second end together with the formed DNA product). In one embodiment, the device does not include a closed loop of PCR fluids (which may be formed, for example, by a reaction tank connected by or through a tube). In one embodiment, the device does not include a closed loop of PCR fluids connected to a valve, which allows the flow of PCR fluids and DNA product, respectively, into separate tubes exiting the device.

[0011] The DNA product can be DNA corresponding to an API (e.g., DNA vaccines) or a DNA template for enzymatic RNA in vitro transcription (hence an intermediate of RNA as API). RNA in vitro transcription refers to a process in which RNA is synthesized in a cell-free system. A DNA template is a DNA molecule that contains a nucleic acid sequence that codes for an RNA sequence to be transcribed in vitro. The DNA template is used as a template for RNA in vitro transcription to produce RNA encoded by the template DNA. The DNA template can be transcribed into mRNA representing the API, which can be formulated into lipid nanoparticles, resulting in encapsulated mRNA, which can be further formulated into a final drug product. Thus, the DNA product can be an active pharmaceutical ingredient (as in DNA vaccines) or the DNA product can be a DNA template that can be transcribed into mRNA as an active pharmaceutical ingredient (hence the DNA is an intermediate, not an API). The DNA product in the context of the present invention is DNA obtained by PCR, i.e. amplified DNA.

[0012] Polymerase chain reaction (PCR) is one of the fastest methods to prepare DNA products. Typically, PCR can be performed in a PCR device depending on laboratory use and scale. It is a technique in molecular biology that synthetically amplifies DNA over several orders of magnitude, generating thousands to millions of copies of a specific DNA sequence. PCR relies on thermal cycling, which includes repeated cycles of heating and cooling for DNA melting and enzymatic replication of the DNA using a thermostable DNA polymerase. The cycle includes a denaturation temperature, an annealing temperature, and an extension temperature. The separate temperatures in the first and second compartments can be one of a denaturation temperature, an annealing temperature, and / or an extension temperature. Specific PCR techniques include, for example, RT-PCR, hot start PCR, long range PCR, RT-qPCR, etc.

[0013] Capillary polymerase chain reaction can be understood as PCR carried out in a tube, pipe, or capillary. The tube or capillary may be very thin or narrow. The tube or capillary may have a very small diameter ranging from about 0.5 to about 50 mm, preferably about 0.5 mm to about 1.5 mm, more preferably about 0.75 to about 1.25 mm. Suitably, the diameter of the capillary or tube is configured to allow rapid and efficient temperature change of the PCR fluid in the tube as the PCR is proceeding, i.e., as the PCR fluid is flowing / proceeding through the tube during PCR. As described herein, the tube passes through separate compartments configured to provide different temperatures, so that the tube and thus the PCR fluid in the tube are exposed to these different temperatures. The tube or capillary, particularly its diameter (or outer dimensions), may be configured to allow capillary action or capillary effect with the PCR fluid. Capillary action or capillary effect is the ability of a liquid to flow in a narrow space without the aid of or against an external force such as gravity. Capillary action or capillary effect can be seen, for example, in the wicking of liquids in thin tubes, in porous materials such as paper, and in some non-porous materials such as sand. It is caused by intermolecular forces between the liquid and the surrounding solid surface. If the diameter of the tube or capillary is small enough, a combination of surface tension forces caused by cohesion within the liquid and adhesive forces between the liquid and the tube or capillary walls act to propel the liquid forward.

[0014] The tube is wound at least from the first compartment to the second compartment and from the second compartment to the first compartment (back). "Wound" can be understood as not forming a line (line segment) that is the shortest distance between two points. Instead, the wound tube can be longer than the shortest distance between two points. The wound tube may extend in a circular, curved, or wavy shape. The reason for winding the tube is that the length of the tube in a particular compartment and a given flow rate determine the time that the PCR liquid is exposed to a specific temperature inside the compartment. This defines how long the PCR liquid in the tube is exposed to a specific temperature required for PCR, such as a denaturation temperature, which may be present, for example, in the first compartment. If the tube passes through the first compartment via the shortest path to reach the second compartment, the length of the tube exposed to the denaturation temperature provided in the first compartment may be too short to ultimately denature the DNA sufficiently. However, if the tube is wound as it passes through the first compartment, the length of the tube exposed to the denaturing temperature provided in the first compartment will be sufficiently long that the PCR solution passing through this section of the tube will be exposed to this temperature for a sufficient period of time to ultimately cause the DNA to be sufficiently denatured.

[0015] It is important to understand that the tube according to the present application is not connected to one (or more) reaction mixture tanks, specifically to one reaction mixture tank (singular or plural). In other words, the tube according to the present application does not allow a recirculation path where the tube starts from a reaction mixture tank, passes through a different tank (at a different temperature) and then returns to the very same reaction mixture tank to provide recirculation. Furthermore, it is important to understand that the tube is not connected to a valve that allows a recirculation path.

[0016] The PCR liquid can be understood as a reaction liquid, which comprises a DNA template for PCR amplification (the DNA template is initially present in small amounts, and this initial DNA template for PCR amplification may preferably be de novo synthesized DNA), one or more DNA primers, dNTPs, a DNA polymerase (e.g., a proofreading DNA polymerase), and a suitable PCR buffer. The PCR liquid is filled into the tube at one end (the tube as a whole has two ends, i.e., an "inlet end" for the starting PCR liquid and an "outlet end" for the resulting amplified DNA product), or pumped, and is guided through the compartments by the tube wound into and through these compartments. The PCR liquid is separated from the areas within the compartments by the tube walls, and thus from, for example, a heat medium (e.g., liquid or gas) or a heat solid filled in the compartments, so that the PCR liquid is not in direct contact with the compartments and the spaces within the compartments, respectively, at any one time. The tube walls allow a temperature exchange between the PCR liquid and the compartment (and thus the temperature in the compartment), e.g., a heat medium or a heat solid in the compartment, which is then used to heat or cool the PCR liquid by a means for regulating the temperature of the tube, e.g., by a heat medium or a heat solid. While the PCR liquid is led through the compartment, the templates for PCR amplification are amplified and a DNA product is generated. This DNA product then exits the tube at a second end of the tube, the "outlet end". Thus, the DNA product does not need to be removed from the reaction mixture tank or any corresponding object in the recirculation system.

[0017] The tube forms a helical stack of tube turns (essentially as exemplarily shown in FIG. 3). One "tube turn" means one "round" of the tube back to the starting first section, i.e., from the first section to the second section and back from the second section to the first section, or preferably, from the first section to the second section, from the second section to the third section, and from the third section to the first section. It is important to understand that the tube returns to the first section, but is not connected to the initial section of the tube in the first section. Instead, the tube continues to be helically wound after making tube turns from the first section through multiple sections back to the first section, eventually forming a helical stack. In other words, the tube does not form a "two-dimensional" loop, where it goes from a first section through a different section and back to the first section, where it is again connected to the first tube, thus forming a loop, but rather a "three-dimensional" spiral, where the tube goes from a first section through a different section and back to the first section without connecting to the first tube. The spiral in the "three-dimensional" may specifically be essentially perpendicular to each turn in the "two-dimensional". This means that "the stacking direction of the spiral stack of tube turns is different from the corrugation direction of the corrugated tube, and preferably essentially perpendicular to the corrugation direction of the corrugated tube". The spiral stack of tube turns allows multiple PCR cycles, with each turn corresponding to one PCR cycle.

[0018] The tube has two separate ends, a "PCR fluid inlet" and a "DNA product outlet", the first end being typically located on the tube before it enters the first compartment (this is where the PCR fluid enters the device) since the denaturation step is the first step of the PCR reaction, and the second end being typically located on the tube after it leaves the second / third compartment (this is where the DNA product exits the device) since the extension step is the last step of the PCR reaction.

[0019] The device for preparing DNA products comprises at least two (tempering) compartments, a first compartment and a second compartment. It is also possible, and typically preferred, to provide a third compartment. More compartments are also possible. A compartment can be understood as an at least partially, preferably completely, closed space, room or container. The compartment may have a volume of at least 10 mL, preferably at least 100 mL, more preferably at least 1 L, for example from about 1 L to about 10 L. The compartment can be sealed and leak-proof. The at least two compartments may form at least two independent or different tanks, for example filled with a heat medium or a heat solid. The at least two compartments may be separated from each other, preferably insulated from each other, to allow separate, different or independent temperature zones in each of the at least two compartments. These temperature zones correspond to the temperature profile of PCR. For the efficiency of PCR, it is particularly important that the "ramp rate", which indicates the temperature change over time, is rapid. In the context of the present invention, the temperature change over time is determined by the flow rate of the PCR liquid and the time required for heat exchange in the compartment.

[0020] One option is to regulate the temperature in each compartment by a heat medium. The heat medium may be liquid or gas. In that case, the heat medium can enter each of the at least two compartments by a respective heat medium inlet and can exit each of the at least two compartments by a respective heat medium outlet in one embodiment. Thus, in this embodiment, each compartment comprises a heat medium inlet and a heat medium outlet. Thus, there can be a flow of the heat medium through each tank in the at least two compartments that is at least partially filled with the heat medium. The heat medium can be understood as a liquid coolant and heat carrier, for example, based on propylene glycol, silicone-based fluids or other synthetic heat mediums, as well as high thermal conductivity media such as water or any mixture of the above liquids. The heat medium can also be understood as a gaseous coolant or gaseous heat carrier. The heat medium can be, for example, Pekasol®. The heat medium can be used to provide, control and / or regulate separate temperatures in the at least two compartments to prepare DNA products by thermal cycling that is part of PCR. The heat media in the separate compartments are configured to allow rapid temperature changes of the tubes, and thus the PCR fluids within the tubes, during PCR. In one embodiment, a compartment does not include an agitation means for mixing the heat media within that compartment. In another embodiment, a compartment includes an agitation means for mixing the heat media within that compartment.

[0021] Another option is to regulate the temperature in each compartment by a thermal solid. In that case, the temperature of the thermal solid is provided and regulated in this embodiment by a heating unit and optionally a cooling unit. In this embodiment, no liquid thermal medium is used, and therefore no thermal medium inlet and outlet are present in the compartment. The thermal solid is a thermally conductive solid, and therefore provides a specific temperature to the compartment and the tube surrounded by the thermal solid, respectively (the thermal solid is configured to allow a rapid temperature change of the tube, and thus of the PCR liquid in the tube, during PCR). The thermal solid can be selected from the group consisting of aluminum, aluminum alloys, or other alloys with high thermal conductivity, copper, bronze, and brass, with aluminum or aluminum alloys being preferred. The thermal solid may have the form of pellets, balls, and / or coarse powder, preferably with a diameter of about 1 to about 3 mm in the case of pellets and balls. It is particularly preferred to use aluminum pellets. The thermal solid may be contained in a liquid or gel to remove residual air in the compartment. Thus, in this configuration, the compartment surrounding the tube (or the part of the compartment formed by the internal compartment walls (30), see FIG. 8) is filled with a thermal solid, as outlined above, and liquid or gel, and is free of air. The heating unit is preferably an electric heating unit. Advantageously, the heating unit is integrated into the stand (or the tube holding means) to allow optimal heating. The electric heating unit in the compartment can be, for example, a heating cartridge or a heating wire or a heating conductor. In addition, it is further possible to cool the compartment (or the part formed by the internal compartment walls (30, see FIG. 8) of the compartment) to provide and / or regulate the required temperature by using a cooling unit, in particular a thermoelectric cooler (i.e. a Peltier element) with a heat pipe connected thereto (which heat pipe extends into the compartment and is therefore also inside the compartment). The thermoelectric cooler can also be used to provide additional heat to the compartment when operating in a heating mode, which is the opposite mode to cooling. When operating in a cooling mode, a ventilator may be present to remove the heat of the thermoelectric cooler.However, the presence of a ventilator is not mandatory, since the heat obtained from the cooling can also be used as additional heating in the (other) compartments of the device. Typically, several independent electronic heating systems are present in each compartment, each of these systems being connected to its own temperature sensor and achieving several independent control circuits.

[0022] The device according to the invention for preparing DNA products allows for flexible adaptation to different conditions, requirements and, for example, amounts of DNA. The device may allow for the preparation of large amounts of DNA (e.g., more than 2 mg per reaction). Furthermore, the preparation of DNA products may be scalable. In particular, the preparation of DNA products may be continuous, and g quantities and essentially unlimited amounts may be produced, since new PCR liquid is continuously added to the "inlet" while the resulting DNA leaves the device through the respective "outlets" of the tubes and the device. The device for preparing DNA products allows for very rapid preparation of DNA products, and thereby also rapid production of pharmaceutical products or intermediates. Also, modification and adaptation of the device for preparing DNA products to a specific DNA product may be very rapid. There may be no need for adjustment or calibration steps. There may be no need for washing steps or they may be lower. For example, the turnover time for vaccine production may be less than about one week.

[0023] The device according to the present invention for preparing DNA products is constructed in a very elegant manner. The device can be dimensioned to be very small. The device for preparing DNA products by capillary PCR is portable, allowing for example transportation to pandemic areas. It may allow for local, decentralized production of intermediates (e.g., DNA templates) of medicines (e.g., DNA vaccines) or medicines (e.g., RNA vaccines).

[0024] The device for preparing DNA products according to the invention can be used for flexible and rapid DNA vaccine production or DNA template production for RNA vaccines, in particular for flexible and rapid RNA vaccine production. The device can be used in particular for the production of DNA templates for mRNA-based vaccines, for example during epidemics and pandemics of infectious diseases, as well as for various cancer diseases, including personalized treatment.

[0025] In one embodiment, a partition is placed between the compartments, isolating them from each other. This can be understood as a partition being placed between the first and second compartment. If there are more compartments, for example three, a partition may be placed between the first and second compartment, another partition may be placed between the second and third compartment, and / or a further partition may be placed between the third and first compartment. As a result, the (adjacent) compartments can be insulated from each other and thus different temperatures can be established and maintained in the (adjacent) compartments. The partition may be implemented by only one wall component, isolating all the compartments by the same wall component, or by two or more wall components. The partition may be a material with low thermal conductivity and good mechanical and temperature resistance. Additionally or partially alternatively, there may be an internal partition wall, which fits more closely to the corrugated tube and thus forms a narrower space surrounding the tube in each compartment. The internal partition walls also provide isolation of the respective compartments while forming a fairly narrow space around the tubes, which is filled with, in particular, a thermal solid. If an internal partition wall is present, the partition does not necessarily exist entirely between the compartments, but may exist only partially, in particular only in the area where the tube passes from one compartment to the next through the partition, as will be explained below.

[0026] Thus, in the most preferred embodiment, the partition comprises a through hole for a tube to extend from one compartment to the next, allowing the tube to pass through the separate compartments. The through hole or channel allows the tube to enter and / or exit the compartment and the temperature zone of the compartment. The through hole may be sealed to improve leak tightness. Also, the through hole may be insulated to improve temperature stability within the (adjacent) compartment.

[0027] In one embodiment, the heat medium inlet and the heat medium outlet, if present, are arranged in at least one compartment, preferably in each compartment, to provide a heat medium flow through the compartment in an essentially opposite direction to the guide direction of the PCR fluid in the tube. This can be understood as meaning that if the guide direction of the PCR fluid is clockwise, the heat medium inlet and the heat medium outlet are arranged to provide a heat medium flow in a counterclockwise direction, and vice versa. Alternatively, if the guide direction of the PCR fluid is from left to right, the heat medium inlet and the heat medium outlet are arranged to provide a heat medium flow in a direction from right to left, and vice versa. The counterflow of the heat medium and the PCR fluid improves the temperature exchange between the heat medium and the PCR fluid, thereby improving the temperature control of the PCR fluid. For an efficient PCR reaction, it is particularly important that the temperature is constant in each of the compartments of the device, and that it promotes a rapid temperature exchange when the tube containing the PCR fluid is led to the next compartment (typically having a different temperature).

[0028] In one embodiment, the heat transfer medium inlet, if present, is located at a lower position in each compartment, and the heat transfer medium outlet, if present, is located at an upper position in each compartment. Such a configuration has the advantage that air bubbles can be more easily removed.

[0029] Alternatively, the heat transfer medium inlets, if present, may be located at an upper position within each compartment, and the heat transfer medium outlets, if present, may be located at a lower position within each compartment. In a particularly preferred embodiment, the device for preparing DNA products by capillary polymerase chain reaction further comprises a third compartment. The tube may be wound at least from the first compartment (through the first compartment) to the second compartment, and from the second compartment (through the second compartment) to the third compartment (through the third compartment). The tube may further be wound from the third compartment to the first compartment, again from the first compartment to the second compartment (through the first compartment), and from the second compartment to the third compartment (through the second compartment). The third compartment also comprises a means for adjusting the temperature within the compartment. In this regard, the third compartment may comprise a heat medium inlet and a heat medium outlet to provide a heat medium flow through the third compartment that is separate from the heat medium flow through the first and second compartments. The separate heat medium flow may be used to adjust a separate temperature in the third compartment that is separate from the temperatures in the first and second compartments. Instead of the heat medium inlet and outlet, an embodiment with a heating unit as described above may be used as a means to adjust the temperature in the compartments. Also, a separate temperature in the third compartment is used to prepare the DNA product based PCR solution. The three temperature zones typically correspond to the PCR temperature profile for denaturation, annealing, and extension.

[0030] The tube wound from compartment to compartment forms a helical stack of tube turns. As defined above, a tube turn is a section where the tube starts from a first compartment and returns to the first compartment, with one tube turn representing one PCR cycle. Such tube turns then form a helix, eventually resulting in a helical stack of tube turns, with each tube turn representing one PCR cycle. In other words, this can be understood as the tube having a shape of a helix, a spiral, or a coil (when viewed in side view). In yet another way, the tube may extend in the form of multiple circles (but the circles are not connected to each other), which makes the device for preparing DNA products by capillary polymerase chain reaction very elegant and space-efficient. One circle may extend from the first compartment to the second compartment, and from the second compartment to the (optional) third compartment. Every circle corresponds to one PCR cycle. The helical extension or stacking direction of the tubes may extend in the so-called z-direction, away from the so-called x,y plane formed by the bottom of the device for preparing DNA products, or the plane formed by the floor or ground. The helical extension of the tubes may extend essentially perpendicular to the bottom, floor or ground of the device, but may also extend at an angle to the bottom, floor or ground of the device.

[0031] In one embodiment, the tube extends in at least one compartment, preferably within each of the compartments, in a wavy or serpentine manner, so that in each compartment the tube is of a certain length, which reflects the incubation time of the PCR fluid inside the tube when the PCR fluid passes through the tube in this compartment, preferably at a predetermined and controlled flow rate. Since this incubation time is typically longer than that achieved if the tube is straight in the compartment, the tube is preferably present in a wavy or serpentine manner in each compartment. This can be understood as the tube (when viewed in top view) having a wave or snake shape in one or all of the compartments. The wave or snake shape or corrugation of the tube may extend in the x- and y-directions of the plane formed by the bottom of the device for preparing DNA products or the plane formed by the floor or ground, i.e. it is understood to relate to a "two-dimensional" form, not including a helical, i.e. "three-dimensional" form. The corrugation of the tube defines the residence time of the PCR fluid in each compartment, and thus the residence time of the PCR fluid in the temperature zone provided by each compartment. Thus, the corrugation of the tube defines the length of the tube in each compartment having a particular temperature, and thus the residence time of the PCR fluid in the temperature zone provided by each compartment.

[0032] In one embodiment, the stacking direction of the spiral stack of tube windings is different from the corrugation direction of the corrugated tube, and is preferably essentially perpendicular to the corrugation direction of the corrugated tube. The angle between the stacking direction of the spiral stack of tube windings and the corrugation direction of the corrugated tube may differ by up to about 90°, for example, less than about 90°, in the range of 45° to 89°. And the stacking direction of the spiral stack of tube windings and the corrugation direction of the corrugated tube may be inclined or oblique to each other. The stacking direction of the spiral stack of tube windings may be inclined or oblique to the bottom of the device for preparing DNA products or to a plane formed by the floor or ground. Additionally or alternatively, the corrugation direction of the corrugated tube may be inclined to the bottom of the device for preparing DNA products or to a plane formed by the floor or ground. And the tube may have a pitch to the bottom of the device for preparing DNA products or to a plane formed by the floor or ground. The pitch may be the same throughout the device for preparing DNA products or throughout one of the compartments, but the pitch may vary throughout the device for preparing DNA products or throughout one of the compartments.

[0033] In one embodiment, the device for preparing DNA products by capillary polymerase chain reaction further comprises at least one stand or tube holding means arranged in one of the compartments to provide a fixing point for the tube and / or a deflection point for the corrugation of the tube. The stand may have a cylindrical shape. Also, the stand may have a T-shape or an L-shape when viewed in top view. Other shapes are also possible. The stand may comprise at least one mounting structure configured to receive and fix the tube (and thus the tube winding and the helical stack of tube windings, respectively). Preferably, the stand comprises a plurality of mounting structures for fixing and holding the tube (and thus the tube winding and the helical stack of tube windings, respectively) in the stand. The mounting structures may be formed, for example, as recesses, as through-holes, as hooks, as slits, or as other means for attaching the tube (and thus the tube winding and the helical stack of tube windings, respectively) to the stand. The tube may be fixed to the mounting structure in a pressure-fit and / or form-fit manner. Additionally or alternatively, the stand may further comprise at least one clamping means for fixing and holding the tube. Preferably, the mounting structure is coated with a soft material, for example including rubber or silicone, to prevent the tube from being scratched, cut or otherwise damaged. Suitably, the stand may be of a material that is chemically resistant to the heat transfer medium used.

[0034] In one embodiment, the tube extends partially around the outer dimension of the stand. This can be understood in a top view as the tube extending less than about 360° around the outer diameter or outer dimension of the stand, but not entirely. The tube may extend from about 45° to about 270° around the outer dimension of the stand. In another embodiment, the tube extends entirely around the outer dimension of the stand. This can be understood in a top view as the tube extending about 360° around the outer dimension or circumference of the stand. In another embodiment, the tube extends more than twice around the outer dimension of the stand. This can be understood in a top view as the tube extending more than about 360° around the outer dimension or circumference of the stand. The tube may extend from about 450° to about 540° around the outer dimension of the stand, meaning that there may be an overlap of the tube around the stand.

[0035] In one embodiment, the stand is exchangeable within the compartment. This can be understood as the stand being feasibly fixed to the compartment, specifically to the bottom plate of the compartment. As a result, the stand can be exchanged, for example, with another stand having another outer dimension (radius and / or height) in order to adapt the device for preparing DNA products to another tube, another purpose, another PCR duration (e.g. to reflect another PCR protocol), another winding radius, another winding pattern, etc.

[0036] In one embodiment, the stand is repositionable within the compartment. This can be understood as the stand can be positioned in a different position or location within the compartment, specifically in a different position relative to the bottom plate of the compartment. The bottom plate may therefore be formed as a perforated plate with (blind) holes into which the stand can be flexibly inserted. As a result, the stand can be repositioned to adapt the device for preparing DNA products to different tubes, different purposes, different PCR durations (e.g. to reflect different PCR protocols), different winding radii, different winding patterns, etc.

[0037] Stands, particularly interchangeable and / or repositionable stands, can be used to define the residence time of the PCR fluids in the compartments, and thereby the residence time of the PCR fluids in the temperature zones provided by the compartments. Thus, the dimensions and position of the stands can define the temperature profile of the PCR, and can be flexibly adjusted to match a desired PCR profile (e.g., to generate different DNA products, e.g., different lengths of DNA products).

[0038] The tube may be continuous throughout the device for preparing DNA products, which can be understood as the tube being the same throughout the device for preparing DNA products. The tube may also be discontinuous throughout the device for preparing DNA products, which can be understood as the tube comprising separate parts, for example having different sizes or diameters in the cross section of the tube. The tube may be interchangeable to adapt the device for preparing DNA products to different PCR or other purposes. For reasons of cleanability, it is preferred that the tube is continuous. Furthermore, the tube is configured to allow corrugation of the tube, for example through a stand, without risk of breakage or bending, which may inhibit the efficiency of PCR. However, in all examples, the tube comprises a defined "inlet" end and a defined "outlet" end, rather than a recirculating or closed tube system with an optional outlet.

[0039] In one embodiment, the tube comprises a PCR liquid inlet, which can be connected to a pump unit for pumping the PCR liquid through the tube. The pumping unit can be a pump. The pumping unit can be an external pumping unit. In one embodiment, the pumping unit is arranged outside the compartment, preferably outside the outer wall surrounding the compartment. The outer wall can be made of a plastic material, for example POM, PEEK, PTFE, but also metals, alloys, compounds, etc., and can be resistant and compatible with the heat and the applied heat medium. The external arrangement can save space in the device for preparing the DNA product and / or allow the use of a pumping unit with a higher output. Preferably, the pumping unit comprises a micro gear pump configured to generate a constant flow rate.

[0040] In one embodiment, the pump unit is configured to provide a flow rate of PCR fluid ranging from about 0.05 mL / min to about 50 mL / min, preferably from about 0.1 mL / min to about 10 mL / min, more preferably from about 0.2 mL / min to about 3 mL / min. Such flow rates can define the residence time of the PCR fluid in the compartments and thus in the temperature zones provided by the compartments. These flow rates can thus define the temperature profile of the PCR. Preferably, the flow rates are constant, e.g., the flow rate fluctuates less than ±10%, preferably less than ±5%.

[0041] In one embodiment, the tube comprises a DNA product outlet for continuous preparation of DNA product. This can be understood as a continuous non-stop preparation of DNA product. Thus, the DNA product outlet can be "always open during production". Such continuous preparation of DNA product enhances continuous and scalable output of large amounts of DNA product. The DNA product outlet may comprise a means consisting of a closing mechanism, for example a shutter, that can be closed when the continuous preparation of DNA product is finished.

[0042] In one embodiment, the tube comprises a closable DNA product outlet for discontinuous preparation of DNA product, which can be understood as discontinuous, batch-by-batch preparation of DNA product. Thus, the DNA product outlet is closable by a closing mechanism, for example a shutter.

[0043] In one embodiment, the device for preparing DNA products by capillary polymerase chain reaction further comprises at least a sensor arranged in at least one of the compartments. This sensor may be a temperature sensor, a flow sensor, a leak sensor, etc., which are configured to detect the temperature, the flow rate, the leak, etc. of the compartment, respectively. The flow sensor may form a closed loop control together with the pumping unit to monitor and control the flow rate outside the device for preparing DNA products or the PCR reactor. At least two temperature sensors may be provided in at least one compartment, preferably in each compartment, e.g., one near the fluid inlet and one near the fluid outlet, for building redundancy. Two leak sensors may be used, one in at least one compartment, preferably in each compartment, and one in the surrounding unit / frame. Further sensors may be arranged outside the compartments, e.g., in the pumping unit, or in the PCR fluid inlet, or in the DNA product outlet.

[0044] The device may also comprise a first part of a proximity sensor, e.g. a magnetic sensor, which finds a counterpart or second part of the proximity sensor, which is outside the device according to the first aspect. With this in mind, this second part of the proximity sensor may in particular be located within the manufacturing device according to the second aspect. When the device is properly installed in the manufacturing device, those two parts of the proximity sensor are connected, preferably signaling a successful installation and / or preferably only then the device can be successfully started and operated. Thus, the proximity sensor including those two parts may also be active during operation, and an alarm may sound and / or operation may be stopped if the two parts of the proximity sensor lose the connection between them.

[0045] In one embodiment, the tube has a diameter (which may roughly correspond to the outer dimensions) in the range of about 0.5 mm to about 50 mm, about 0.5 mm to about 40 mm, preferably about 0.5 mm to about 20 mm, preferably about 0.5 mm to about 4 mm, more preferably about 0.5 mm to about 1.5 mm, and even more preferably about 0.75 mm to about 1.25 mm. These dimensions may allow for rapid and efficient temperature exchange ("ramping") between the heat transfer medium and the PCR fluid.

[0046] The tube may be made of a plastic material, such as PEEK, ETFE, or PTFE, but also of metals, alloys, compounds, etc. Advantageously, the tube may be made of a material that is biocompatible and process compatible (e.g., no leakage of compounds into the DNA product, no oxidation / decomposition of materials, low binding of PCR compounds to the surface to minimize risk of exfoliation, etc.). A preferred material may be PEEK.

[0047] In one embodiment, the tube has a length between the tube inlet and the tube outlet in the range of about 5 m to about 300 m, preferably about 10 m to about 200 m, more preferably about 25 m to about 50 m. The tube can have a length of, for example, about 100 m (e.g., about 97 m). Each section of the tube is contained in a separate compartment, and these dimensions can define the residence time of the PCR fluid within the compartment, and thereby the residence time of the PCR fluid in the temperature zone provided by the compartment. These sections of tube length can thus define the temperature profile of the PCR.

[0048] In one embodiment, the tube is sized to accommodate PCR fluid in the range of about 10 mL to about 1 L, preferably about 20 mL to about 250 mL, more preferably about 30 mL to about 150 mL. The tube may, for example, be sized to accommodate PCR fluid in the range of about 80 mL.

[0049] In one embodiment, the spiral stack of tube windings comprises about 10 to about 50 windings or circles or layers, preferably about 15 to about 40, more preferably about 20 to about 30. The number of windings corresponds to the number of PCR cycles.

[0050] By way of example only, the tube may have a total length of about 15 m, including a spiral stack (or cycle) of about 25 tube turns. In that particular embodiment, in one spiral stack (or cycle), the tube may be guided through a first section (100 cm), followed by a second section (100 cm), followed by a third section (400 cm).

[0051] In one embodiment, the device is configured to prepare about 1 mg or more, for example, up to 5 g, preferably in the range of about 1 mg to about 30 mg, more preferably in the range of about 15 mg to about 25 mg of DNA product per reaction.

[0052] In one embodiment, the device is configured to prepare DNA product in the range of about 2 μg / min to 2000 μg / min, preferably in the range of about 5 μg / min to 1500 μg / min, and more preferably in the range of about 10 μg / min to 1000 μg / min.

[0053] In one embodiment, the device is configured to prepare a range of about 25 μg to about 200 μg of DNA product per mL of PCR solution, preferably about 50 μg to about 150 μg per mL of PCR solution, and more preferably about 75 μg to about 125 μg per mL of PCR solution.

[0054] In one embodiment, the first compartment is configured to have a temperature in the range of about 85°C to about 105°C, preferably about 98°C. In one embodiment, the second compartment is configured to have a temperature in the range of about 45°C to about 72°C, preferably about 60°C to about 72°C.

[0055] In one embodiment, the optional third compartment is configured to have a temperature in the range of about 65°C to about 75°C, preferably about 72°C. By way of example only, the tube may have a total length of about 15 m, including a spiral stack (or cycle) of about 25 tube turns. In that particular embodiment, in one spiral stack (or cycle), the tube may be guided through a first section (100 cm; temperature about 85° C. to about 105° C.), followed by a second section (100 cm; temperature about 45° C. to about 72° C.), followed by a third section (400 cm; temperature about 65° C. to about 75° C.). The residence time in each section may be adjusted by the PCR flow rate and the diameter of the tube.

[0056] Those skilled in the art will understand that different DNA products (e.g., different sizes and / or sequences) may require different PCR conditions (e.g., due to DNA primer annealing temperature or extension time, etc.). By adjusting the length of the tubes, the length of the tubes in the separate compartments, the number of spiral stacks (and thus the number of PCR cycles), the flow rate, the diameter of the tubes, and / or the temperature in the compartments, different PCR programs suitable for various different DNA products can be realized. Thus, the device for preparing DNA products by capillary polymerase chain reaction can be easily adapted and adjusted to enable the preparation production of DNA products of various sizes and sequences.

[0057] In a second aspect, the present invention is directed to a pharmaceutical manufacturing device comprising the device for preparation of a DNA product by capillary polymerase chain reaction of the first aspect. In one embodiment, the pharmaceutical production device comprises a device for preparing a DNA product by capillary polymerase chain reaction as described in the first aspect and at least one further function (e.g. additional chamber, additional unit, housing, etc.) In a preferred embodiment, the pharmaceutical production device according to the second aspect comprises a processing chamber, a technology chamber and a device for preparing a DNA product by capillary polymerase chain reaction as described in the first aspect.

[0058] Description of the Manufacturing Device and Its Preferred Embodiments A pharmaceutical manufacturing device can generally be understood as a closed housing having different chambers for different purposes.

[0059] The processing chamber is suitable for and is used to produce pharmaceutical products. The processing chamber may also be referred to as the "wet part" of the manufacturing device, in the sense that essentially all wet processing during the manufacturing process takes place in the processing chamber. By "wet processing" is meant, for example, the provision of buffers and / or reactants and / or products to and from containers or to and from chromatography columns, the withdrawal of waste liquids or waste gases etc., and the connection (for example in the form of flexible tubes) of such buffers and / or reactants and / or products. Such wet processing should ideally be separated from all sources of technical media supply (in particular from electrical supplies such as power cables) in order to make impossible or at least to minimize accidents due to leakage of liquids in the processing chamber and possible effects, in particular due to voltage. In the unlikely event of an accident due to leakage in the processing chamber, safety is increased due to the aforementioned separation, and the processing chamber only needs to be cleaned. The device according to the first aspect is located in the processing chamber.

[0060] The technological chamber is suitable and used to provide technical support for housing devices that are not in direct contact with the process medium, such as, for example, pumps, motors, mixers, processors, etc. The technological chamber is typically suitable and used to provide (for example to house) a technical media source, in particular a power supply, for example for devices / units that are located in the process chamber and are in direct contact with the process medium. The technological chamber is sometimes also called the "dry part" of the manufacturing device, in the sense that essentially no wet processing as defined above takes place in the technological chamber. As mentioned above, this provides, among other things, an increased safety. Since the cleanliness level required in the technological chamber is lower than in the process chamber, the technological chamber is sometimes also said to be "less clean" than the process chamber. The technical media source for the device according to the first aspect is located in the technological chamber.

[0061] The chambers may be separated by a separating element, which is plate-like, in particular when the separating element is viewed from the processing chamber. Thus, in order not to disturb the gas flow, as outlined below, the separating element may comprise at least one appendage with an opening towards the processing chamber and an extension in the direction of the technological chamber, in which an apparatus in contact with the processing medium, in particular a device according to the first aspect, may be accommodated.

[0062] The manufacturing device, comprising a device for preparing DNA products by capillary polymerase chain reaction, preferably further comprises a control unit configured to control the gas flow through the processing chamber as a gas shower. The gas flow may be provided by a flow unit. The flow unit may be at least a pump or an HVAC system, also sometimes called a fan system. The flow unit may be located outside the housing of the manufacturing device (external flow unit) or preferably within the housing of the manufacturing device (internal flow unit). The flow unit preferably comprises a filter unit (also sometimes called a "fan filter unit" and preferably comprising a HEPA filter as described below). The flow unit may preferably be configured to provide a variable amount of gas. The gas may be clean air, air with additives, an inert gas, or any other gas. The control unit may be understood as any control device for the gas flow, such as, for example, a valve or a processor. The control unit may be configured to regulate different pressures in the gas flow. The control unit may be configured to control the gas flow through the processing chamber to provide a positive pressure in the processing chamber. A positive pressure in a processing chamber can be understood as an overpressure relative to the environment outside the housing and / or as a pressure higher than in the technology chamber. A positive pressure in a processing chamber can protect open processing steps from particulate contamination.

[0063] The control unit is configured to control the gas flow through the treatment chamber to provide a gas shower through the treatment chamber. The gas shower may fall from a higher height in the treatment chamber to a lower height in the treatment chamber. The gas shower in the treatment chamber may protect the open treatment process from particulate contamination. In an embodiment, the gas shower is laminar, which may be understood as being essentially similar to laminar flow, or essentially unidirectional, or essentially non-turbulent. The laminar gas shower or gas flow may be directed downwards or horizontally, preferably with a constant flow, towards the bottom of the treatment chamber. The laminar gas shower may allow to reduce and maintain particulates such as dust, airborne organisms, or evaporated particles to extremely low levels without introducing turbulence and / or without contaminating the manufacturing procedures in the respective housing or chamber. The devices attached in the area of ​​the gas shower may be firmly attached or covered or encapsulated to the separation element or treatment chamber wall to reduce or eliminate the turbulence or disturbance of the gas shower. The device may also be "hidden" in an attachment to the separation element or in an attachment to the processing chamber wall to reduce or eliminate turbulence or disturbance of the gas shower. In one embodiment, the velocity of the gas shower ranges from about 0.2 to about 0.6 m / s, preferably from about 0.36 to about 0.54 m / s. This velocity may provide a good balance between reducing and keeping particulates to very low levels and not disturbing e.g. manufacturing procedures in the processing chamber.

[0064] The pharmaceutical production device according to the second aspect is flexible in adapting to different conditions, requirements and, for example, quantities of pharmaceutical products, since the process is operated in a continuous or semi-continuous mode. The production device allows for the preparation of large quantities of pharmaceutical products. Furthermore, the production of pharmaceutical products is generally scalable.

[0065] The manufacturing device may provide a closed (biosynthetic) process from raw materials (e.g., dNTPs, DNA primers, DNA templates) to the final product. The manufacturing device may be understood as manufacturing-in-a-box. The manufacturing device of the present invention may be dimensioned to be very small. The manufacturing device of the present invention has a small footprint and may form a manufacturing object with a small footprint. The footprint may be, for example, in the range of about 100 x about 100 x about 200 cm so that it can be easily transported and stored in a typical shipping container of normal size. The manufacturing device is therefore portable, allowing for transportation to, for example, pandemic-affected areas. The manufacturing device thereby generally allows for localized decentralized production of pharmaceuticals (e.g., DNA vaccines, DNA templates).

[0066] The manufacturing device generally allows for very rapid production of pharmaceutical products. The changeover and preparation of the manufacturing device to a particular pharmaceutical product, as well as the production of the particular pharmaceutical product itself, can be very rapid. There may be no need for adjustment or calibration steps. There may be no or less need for cleaning steps. For example, the turnover time for vaccine production can be less than about one week.

[0067] The manufacturing device of the present invention is generally very easy to clean, which can reduce the cleaning effort. This is because of the clean and hygienic design from the beginning. The manufacturing device of the present invention may comprise an aluminum housing, which may be pre-treated, for example, by anodizing and passivation, antibacterial surface finish, for example with silver ion-containing colors, electrolytic polishing surfaces, use of compatible materials throughout the manufacturing device, accessibility of many or all parts of the manufacturing device for cleaning purposes, smooth and seamless cleanable exposed surfaces, matched internal diameters and shapes for optimal automatic cleaning-in-place (CIP) procedures and / or antibacterial cleaning, sealing of crevices or hollow areas, reduction or cessation of any accumulation of condensation and sediment, etc. In particular, the housing, chamber(s), and / or separation elements may be made of aluminum, preferably surface-treated aluminum that allows CIP. Additionally or alternatively, the exterior and / or interior surfaces of the housing, in particular the surfaces within the chamber(s), may have an antibacterial surface finish. Furthermore, the equipment within the housing, in particular the equipment within the processing chamber, may be encapsulated to provide a smooth surface. Furthermore, moving parts or parts directing the technical media may be encapsulated for safety and to allow easy cleaning. Additionally or alternatively, the devices in the housing, in particular the devices in the processing chamber, may be embedded in the walls, in particular in separating elements, and in the best case flush with the respective walls or surfaces. These options improve the cleanability and maintain the gas flow without interruption or undesirable bypass by non-coplanar elements in the processing chamber.

[0068] The manufacturing device can be operated under GMP (Guidelines for Good Manufacturing Practice) compliant conditions. At least the processing chamber, preferably the manufacturing device, can be operated according to the requirements of the EU Guidelines for Good Manufacturing Practice, Annex 1. In one embodiment, the GMP requirements are the requirements of the EU Guidelines to Good Manufacturing Practice Medicinal Products for Human and Veterinary Use, Annex 1, Manufacture of Sterile Medicinal Products (corrected version), European Commission, Brussels, 25 November 2008 (revised). In another embodiment, the GMP requirements are the requirements of the FDA (2004) Guidance for Industry, Sterile Drug Products Produced by Aseptic Processing-Current Good Manufacturing Practice, USDepartment of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER) Center for Biologics Evaluation and Research (CBER) Office of Regulatory Affairs (ORA) Pharmaceutical CGMPs. In another embodiment, the manufacturing device may be operated in compliance with CE, ISO, or GAMP5 standards. The manufacturing device may enable GMP-compliant in-box manufacturing. GMP-compliant conditions may include compliant machine control and data logging, such as automatically logging and reporting critical process parameters and / or quality attributes, such as pressure, humidity, temperature, UV absorption, total organic carbon, infrared absorption spectrum, flow rate, power consumption, events that cause errors (open doors, leaks, shortcuts, pressure loss), etc.

[0069] In one embodiment, the manufacturing device comprising a device for preparing a DNA product by capillary polymerase chain reaction further comprises a separation element separating the processing chamber from the technical chamber. The processing chamber is suitable for and may be used to manufacture pharmaceutical products. The technical chamber is suitable for and may be used to accommodate devices such as pumps, motors, mixers, processors, etc. The separation element is in the form of a plate. The separation element may be used to mount devices to the separation element. The mounted devices may be coated to reduce turbulence or disturbance of the gas flow, prevent precipitation, and / or facilitate cleaning. The separation element may extend (only) between the processing chamber and the technical chamber, or may extend at least partially or completely through the housing. The separation element may also extend outside the housing to separate a part of the surrounding environment, such as, for example, a surrounding clean room.

[0070] In one embodiment, the separation element comprises an attachment with an opening towards the processing chamber. The attachment may protrude in the direction of the technology chamber or have an extension in the direction of the technology chamber, which extension serves as an extended processing chamber part. The separation element and the attachment can be understood as a plate with blind holes. The attachment may be used to at least partially or completely accommodate the device. The device for preparing the DNA product (alternatively sometimes called a "PCR reactor") may be insertable into the attachment. Thus, the device for preparing the DNA product or the PCR reactor may comprise or be accommodated in a box to facilitate insertion into the attachment. For example, a device such as a PCR reactor may not intrude into the processing chamber or may only intrude slightly, but still be considered to be located in the processing chamber. This allows the available space in the processing chamber to be increased without increasing the length of the processing chamber. Furthermore, the device may allow the plug-and-play of different devices, for example for another or new PCR process (e.g., different PCR reactors reflecting different PCR programs by having different tube lengths).

[0071] In one embodiment, to facilitate handling of the device for preparing a DNA product, at least a handle is located on the front of the device for preparing a DNA product for handling the device for preparing a DNA product in and / or out of the attachment.

[0072] In one embodiment, the attachment comprises an exchange rail system that guides and facilitates the exchange of the device for preparing DNA products with another device. The exchange rail system may comprise an exchange rail arranged on the attachment or on the device for preparing DNA products (specifically on its box). The device for preparing DNA products may further comprise a handling device, such as, for example, a trolley, a lift, etc., to enable and / or facilitate the exchange of the equipment, for example, from the processing chamber side.

[0073] In one embodiment, when the device for preparing DNA products is inserted into the attachment, the front of the device for preparing DNA products or the front of its box ends essentially flush with the opening of the attachment into the processing chamber, so that the gas flow, in particular the gas shower, is not or is less disturbed and / or the precipitation surface is reduced.

[0074] In one embodiment, the inlet for the PCR liquid and / or the outlet for the DNA product are located on the front side of the device for preparing DNA products or its box, while in another embodiment, the inlet for the heat medium and / or the outlet for the heat medium are located on the rear plate of the device for preparing DNA products, opposite to the front side of the device for preparing DNA products.

[0075] In another, less preferred embodiment, the reverse is also true: the appendage may have an opening towards the technology chamber and an extension in the direction of the processing chamber. In all cases, the appendage may be closable by a cover.

[0076] In one embodiment, the manufacturing device, which comprises a device for preparing a DNA product by capillary polymerase chain reaction, further comprises a leak sensor disposed in an outer attachment of the device for preparing a DNA product. The leak sensor can improve the safety of the device for preparing a DNA product. As described above in the first aspect, the manufacturing device can also comprise a second part of the proximity sensor, the first part being located in the device of the first aspect. When the device is properly installed in the manufacturing device, those two parts of the proximity sensor are connected, preferably signaling successful installation and / or preferably allowing the device to be operated and started successfully only then. Thus, the proximity sensor including those two parts may also be active during operation, and an alarm may sound and / or operation may be stopped if the two parts of the proximity sensor lose the connection between them.

[0077] In one embodiment, the pump unit for pumping PCR liquid through the tube of the device for preparing DNA product is arranged outside the device of the first aspect. The pump unit is preferably arranged in the processing chamber of the device for manufacturing pharmaceutical products. Alternatively, the pump unit may be an external pump unit. The external arrangement can save space in the manufacturing device and / or allow the use of a pump unit with a high output. The pump unit may be a pump.

[0078] In one embodiment, the manufacturing device comprising a device for preparing a DNA product by capillary polymerase chain reaction further comprises a coupling unit arranged in a casing of the pharmaceutical manufacturing device and configured to couple the pharmaceutical manufacturing device to another manufacturing device. The coupling unit can enable and facilitate coupling of two or more manufacturing devices. The resulting manufacturing module is described in a third aspect of the present specification.

[0079] The pharmaceutical production device can be understood as a closed housing having a technological chamber and a processing chamber. In one embodiment, the housing is sealed against the environment. In the same or another embodiment, the processing chamber is sealed against the technological chamber. Throughout this application, the term "sealed" can be understood as complying with at least IP64. The IP code is based on the degree of protection provided by the enclosure (IP code) (IEC 60529:1989+A1:1999+A2:2013); the German version EN 60529:1991+A1:2000+A2:2013 and / or ISO 20653:2013-02 Road vehicles - Degree of protection (IP code) - Protection of electrical equipment against foreign objects, water and access. The term "sealed" can be understood as protection against dust and splash water. "Dustproof" can be understood as no ingress of dust and complete protection against contact. For the test, a vacuum is typically applied and the test time is up to 8 hours depending on the airflow, details of which can be found in the standards cited above. "Protection against splashing water" can be understood as water being splashed onto the enclosure from any direction without harmful effects using a vibrating instrument for 10 minutes or an unshielded spray nozzle for a minimum of 5 minutes, details of which can be found in the standards cited above. A sealed housing or sealed treatment chamber can have the advantage of being particularly clean, meaning having and maintaining low levels of particulates such as dust, airborne organisms or evaporative particles.

[0080] In one embodiment, the treatment chamber is a clean room and / or configured to be at least a grade D room according to the above mentioned EU guidelines on good manufacturing practice for pharmaceuticals, Annex 1. Preferably, the treatment chamber is configured to be at least a grade C room. More preferably, the treatment chamber is configured to be at least a grade B room. Even more preferably, the treatment chamber is configured to be a grade A room. Grades A-D rooms may offer advantages, specifically they are designed to be clean, meaning they have and maintain very low levels of particulates such as dust, airborne organisms, or evaporated particles. The cleanliness can be quantified by the number of particles per cubic meter on a given molecular scale. The characteristics of the A-D grades can be found in Table 1, inserted below. Thereby, the treatment chambers can be classified according to the number and size of particles allowed per volume of gas. The number and size of particles can be measured by particle counting, either based on light scattering, light obscuration, or direct imaging. A high intensity light source is used to illuminate the particles as they pass through the detection chamber. Particles pass through a light source (typically a laser or halogen light) and, with light scattering, the redirected light is detected by a photodetector. With direct imaging, a halogen light back-illuminates the particles within the cell while the passing particles are recorded with a high-resolution, high-magnification camera. The recorded video is then analyzed by computer software to measure particle attributes. With light blocking (obscuration), the loss of light is detected. The amplitude of the scattered or blocked light is measured and the particles are counted and tabulated into standardized counting bins. Direct imaging particle counting employs the use of high-resolution cameras and lights to detect particles. Vision-based particle size measurement units capture two-dimensional images that are analyzed by computer software to obtain particle size measurements.

[0081] [Table 1]

[0082] Comparison of rooms graded A to D according to EU guidelines for good manufacturing practice for medicinal products, annex 1, and the FED-STD-209e(1992)Federal Standard 209-e Airborn Particulate Cleanliness Classes in Cleanrooms and Clean Zones, the ISO(1999)International Standard 14644-1:Cleanrooms and associated controlled environments-Part 1:Certification of Air Cleanliness, International Organisation for Standardisation, Switzerland, and ISO(2003)International Standard ISO 14698-2:2003 Cleanrooms and associated controlled environments-Biocontamination control-Part 2:Evaluation and interpretation of biocontamination data are shown in insert table 2 below.

[0083] [Table 2]

[0084] In one embodiment, the control unit is configured to control the gas flow to provide a gas flow from the processing chamber into the technology chamber. This can be understood as gas flowing from the processing chamber to the technology chamber and into the technology chamber. In the same or another embodiment, both chambers are arranged such that the gas flow in the technology chamber is parallel to but opposite to the gas flow in the processing chamber. Preferably, there is a gas flow in the housing to the processing chamber, into the processing chamber, through the processing chamber, out of the processing chamber, to the technology chamber, into the technology chamber, through the technology chamber, out of the technology chamber, but not all steps are necessary.

[0085] The gas may enter the housing before entering the processing chamber. The gas may leave the housing after leaving the technological chamber. The manufacturing device may therefore further comprise at least one duct passing through the housing for the passage of the gas flow between the housing and the environment. The duct may be an inlet and / or an outlet for the gas. Preferably, the duct is sealed against the environment. As mentioned above, the term "sealed" can be understood to be at least IP64 compliant, as well as dustproof and protected against splashes of water. The seal may be a silicone seal.

[0086] In one embodiment, the housing comprises passages for gas flow from the processing chamber to the technology chamber. A passage can be understood as a channel for the gas. The passage or channel may be provided as a duct or as an intermediate chamber arranged outside and between the processing chamber and the technology chamber (downstream). The passage or channel may also cross the separation element as a sealed conduit or a sealed through-hole. As mentioned above, the term "sealed" can be understood as being dust-proof and protected from splashing water as well as complying with at least IP64. A sealed conduit may have the advantage of maintaining a low level of particulates. The sealed conduit may also comprise a filter to further reduce the level of particulates and / or a valve unit to control the gas flow. The valve unit may be a valve, preferably a throttle valve.

[0087] In one embodiment, the control unit is configured to control the gas flow to provide a pressure difference between the processing chamber and the environment in the range of about 5 to about 100 Pa, preferably about 10 to about 20 Pa or about 15 Pa. The pressure difference may preferably be at least about 15 Pa. This can be understood as a higher pressure in the processing chamber than in the environment including the technology chamber. At least the pressure in the processing chamber can be higher than atmospheric pressure. The higher pressure in the processing chamber makes it less likely that particles can enter the processing chamber and contaminate the processing chamber. In one embodiment, the control unit is configured to control the gas flow to provide a pressure difference between the technology chamber and the environment in the range of about -5 to about -100 Pa, preferably about -10 to about -20 Pa. The pressure difference may preferably be at least about -15 Pa. In one embodiment, the control unit is configured to control the gas flow to provide a pressure difference between the processing chamber and the technology chamber in the range of about 10 to about 200 Pa, preferably about 20 to about 40 Pa. The pressure difference may preferably be at least about 30 Pa.

[0088] In one embodiment, the pressure in the processing chamber is higher than the pressure in the technology chamber. This is met either when the pressure in the processing chamber and the pressure in the technology chamber are higher than atmospheric pressure, or when the pressure in the processing chamber is higher than atmospheric pressure and the pressure in the technology chamber is the same as atmospheric pressure, or when the pressure in the processing chamber is higher than atmospheric pressure and the pressure in the technology chamber is lower than atmospheric pressure. In other words, in one embodiment, the control unit is configured to control the gas flow to create a negative pressure in the technology chamber with respect to the environment.

[0089] In one embodiment, the pharmaceutical manufacturing device further comprises a fluid collection tray for collecting leaked fluids. The tray can be placed at the bottom of the housing and / or in the passage from the processing chamber to the technical chamber. This is beneficial because it allows humidity to be collected without harming the product or the equipment placed in the chamber. The humidity can be implemented intentionally or can result from gas expansion in the technical chamber.

[0090] In one embodiment, the pharmaceutical manufacturing device further comprises a sensor unit comprising at least one of the group consisting of a flow sensor, a pressure sensor, a temperature sensor, a humidity sensor and a leak sensor. The sensor unit may be arranged in the housing, for example in the separation element, in the process chamber, in the technology chamber, in an intermediate chamber between them or as part of the technology chamber, in a conduit passing through the separation element, at the bottom of the housing, etc. The sensor unit may make it possible to control conditions taking into account, for example, flow rate, pressure, temperature, humidity, leaks, etc. Thus, the sensor unit may comprise at least one of the group consisting of a flow sensor, an air pressure sensor, a temperature sensor, a humidity sensor, a microorganism sensor, a particulate sensor, an organic sensor and a leak sensor.

[0091] In one embodiment, the pharmaceutical manufacturing device further comprises a coupling unit arranged on the outer wall of the housing and configured to couple the manufacturing device to another manufacturing device. The other manufacturing device may be similar to the pharmaceutical manufacturing device described above or may be a different device. The coupling unit may allow mechanical coupling of the at least two manufacturing devices. The coupling unit may allow fluidic communication and coupling of the at least two manufacturing devices. The coupling unit may allow data communication and coupling of the at least two manufacturing devices. The fluidic communication may specifically include transporting the pharmaceutical from one manufacturing device to another manufacturing device, for example transporting a DNA template after purification and / or filtration in the first manufacturing device from this first manufacturing device to a second manufacturing device, where the DNA template may then be used in an RNA generation unit included in the second manufacturing device.

[0092] The binding unit is also preferably configured to include a quality control unit, which is suitable for analyzing samples obtained and / or provided from the communication, preferably the fluid communication, between the devices. Thus, as exemplified above, for example, when a DNA template is transferred from a first device to a second device, the sample can be taken and analyzed by a quality control unit, which is preferably located in or at the binding unit.

[0093] A pharmaceutical product can be understood as an active pharmaceutical ingredient (e.g., a DNA vaccine) or any precursor or intermediate thereof (e.g., a DNA template for RNA in vitro transcription). Thus, a pharmaceutical product can be an active pharmaceutical ingredient used in a medicine and administered, for example, to a human subject, for the treatment or prevention of a disease, or an intermediate thereof.

[0094] Production of DNA-based medicines In particular, it may be desirable to produce DNA-based pharmaceuticals, such as DNA vaccines (e.g., lipid nanoparticle-encapsulated DNA vaccines). Essentially, once the sequence of the target, e.g., the sequence of a viral protein (e.g., spike protein in the case of SARS-CoV2), is known, the production of the DNA vaccine can begin. The process then typically follows the steps of i) de novo synthesis of DNA with the desired sequence; ii) DNA amplification by a device according to the first aspect of the present application, optionally followed by DNA modification; iii) encapsulation of the DNA in a formulation; and iv) product filling and finishing, each step building on the previous step. Of course, typical purification and filtration steps will also be performed between the aforementioned steps, if applicable.

[0095] In this embodiment, the manufacturing device of the pharmaceutical product according to the present application further comprises at least one of the group consisting of a processing medium supply unit, a mixing unit, a de novo DNA synthesis unit, a formulation unit, a purification unit, a filtration unit, a DNA modification unit, a filling and finishing unit, and combinations thereof. The required units may be present either in one manufacturing device or in at least two manufacturing devices that are combined to provide a manufacturing module as described herein in the third aspect. In a particularly preferred embodiment, each unit comprises a technical medium source located in the technical chamber, a processing medium source and / or a device configured for contacting the processing medium located in the processing chamber, and a sealing through-hole located in the separation element configured to connect the technical medium source and the device configured for contacting the processing medium. The unit may further comprise a waste outlet, for example for the recovery of washing buffers or by-products.

[0096] In a preferred embodiment, the pharmaceutical production device comprising the device according to the first aspect further comprises a processing medium supply unit, a mixing unit, a purification unit, an optional DNA modification unit, and a filtration unit, preferably connected in the following order: i) processing medium supply unit, ii) mixing unit, iii) device according to the first aspect, optionally followed by a DNA modification unit, iv) purification unit, and v) filtration unit, the device being configured to amplify DNA, and the amplified DNA is optionally modified. The DNA template for the device according to the first aspect can be generated as de novo synthesized DNA in another production device connected to the production device according to this embodiment. The amplified DNA can be used in a separate production device connected to the device of this embodiment and comprising a DNA formulation unit.

[0097] Production of RNA-based medicines In particular, it may be desirable to produce RNA-based pharmaceuticals, such as RNA vaccines (e.g., lipid nanoparticle-encapsulated RNA vaccines). Essentially, once the sequence of the target is known, e.g., the sequence of a viral protein (e.g., spike protein in the case of SARS-CoV2), the production of the RNA vaccine can begin. The process then typically follows the steps of i) de novo synthesis of DNA with the desired sequence; ii) generation of a DNA template by a device according to the first aspect of the present application, optionally followed by DNA modification; iii) RNA generation; iv) encapsulation of the RNA in a formulation; and v) product filling and finishing, each step building on the previous step. Of course, typical purification and filtration steps will also be performed between the aforementioned steps, if applicable.

[0098] In this embodiment, the manufacturing device of the pharmaceutical product according to the present application further comprises at least one of the group consisting of a processing medium supply unit, a mixing unit, a de novo DNA synthesis unit, a DNA modification unit, an RNA production unit, a formulation unit, a purification unit, a filtration unit, a filling and finishing unit, and combinations thereof. The required units may be present either in one manufacturing device or in at least two manufacturing devices that are combined to provide a manufacturing module as described herein in the third aspect. In a particularly preferred embodiment, each unit comprises a technical medium source located in the technical chamber, a processing medium source and / or a device configured for contacting the processing medium located in the processing chamber, and a sealing through-hole located in the separation element configured to connect the technical medium source and the device configured for contacting the processing medium. The unit may further comprise a waste outlet, for example for the recovery of washing buffers or by-products.

[0099] In a preferred embodiment, a pharmaceutical production device comprising the device according to the first aspect further comprises a processing medium supply unit, a mixing unit, a purification unit, an optional DNA modification unit, and a filtration unit, preferably connected in the following order: i) processing medium supply unit, ii) mixing unit, iii) device according to the first aspect, optionally followed by a DNA modification unit, iv) purification unit, and v) filtration unit, and the device is configured to produce a template DNA, and the amplified DNA is optionally modified. This template DNA can be used in a separate production device comprising an RNA production unit, connected to the device of this embodiment.

[0100] The above disclosed units are described in further detail in the third aspect below. For both the production of DNA-based pharmaceuticals and the production of RNA-based pharmaceuticals, the device according to the first aspect of the invention (included in the manufacturing device according to the second aspect) will of course be adapted to the amplification of a specific DNA to be amplified, in particular by having tubes wound and stacked through the compartments such that the resulting PCR conditions and cycles are optimal for this specific DNA to be amplified. When the amplification of a second, different DNA becomes necessary, the first device or PCR reactor can be easily exchanged in the manufacturing device for another device or PCR reactor according to the first aspect, which is adapted to the amplification of a second DNA to be amplified, in particular by having tubes wound and stacked through the compartments such that the resulting PCR conditions and cycles are optimal for this second DNA to be amplified. Thus, the manufacturing device can be easily equipped (or "loaded") with a PCR reactor suitable for the amplification of a specific DNA of interest, each of which is sometimes referred to as a "plug and play" system.

[0101] In a third aspect, the present invention is directed to a pharmaceutical production module. The production module comprises at least two production devices, including a device according to the second aspect. The at least two production devices are coupled to each other. The coupling can allow for mechanical coupling of the at least two production devices. The coupling can allow for fluidic communication and coupling of the at least two production devices. The coupling can allow for data communication and coupling of the at least two production devices. The fluidic communication specifically includes transporting the pharmaceutical from one device to another device, for example transporting a DNA template after purification and / or filtration in the first device from the first device to the second device, where the DNA template can then be used in an RNA production unit contained in the second device.

[0102] The manufacturing module may specifically be for the production of DNA templates and may comprise a process medium supply unit, a mixing unit, a device according to the first aspect (sometimes called a "DNA template generation unit"), an optional DNA modification unit, a purification unit, and a filtration unit, the units being preferably connected in the order listed, and the modules being configured to produce template DNA. The amplification process (or "DNA template generation process") is carried out using the device according to the first aspect.

[0103] The manufacturing module comprises a manufacturing device for a pharmaceutical product according to the second aspect and may further comprise at least one manufacturing device selected from the group consisting of a manufacturing device comprising a process medium supply unit and a mixing unit, a manufacturing device comprising a purification unit, and a manufacturing device comprising a filtration unit.

[0104] In one embodiment, the manufacturing module comprises (i) a first manufacturing device comprising a processing medium supply unit and a mixing unit, (ii) a second manufacturing device according to the second aspect, (iii) a third manufacturing device comprising a purification unit, and (iv) a fourth manufacturing device comprising a filtration unit, preferably connected in the listed order. Such a manufacturing module is configured to produce a DNA template from DNA (optionally de novo synthesized DNA) provided in the form of a processing medium source, preferably in a PCR solution. In other words, such a manufacturing module is configured to produce a large amount of template DNA from a small amount of DNA (which may be de novo synthesized DNA and may be produced by a preceding manufacturing module). This template DNA may be optionally modified by a DNA modification unit included in a further manufacturing device.

[0105] This template DNA can in particular be used for producing RNA, preferably mRNA, by RNA in vitro transcription. Thus, either in the combined or separate production module, an RNA generation unit (for transcribing the resulting DNA into RNA) can be included.

[0106] In one embodiment, the manufacturing module comprises a processing medium supplying unit, a mixing unit, a de novo DNA synthesis unit, a device according to the first aspect (sometimes referred to as a "DNA template generation unit"), optionally a DNA modification unit, a purification unit, a filtration unit, a formulation unit, and an RNA generation unit, preferably arranged in the following order: i) processing medium supplying unit, ii) mixing unit, iii) de novo DNA synthesis unit, iv) purification unit, v) filtration unit, vi) processing medium supplying unit, vii) mixing unit, viii) device according to the first aspect optionally in combination with a DNA modification unit, ix) purification unit, x) filtration unit, xi) processing medium supplying unit, xii) mixing unit, xiii) RNA generation unit, xiv) purification unit, xv) filtration unit, xvi) processing medium supplying unit, xvii) formulation unit, xviii) purification unit, and xix) filtration unit, and the modules are preferably configured to produce formulated RNA, preferably formulated mRNA, more preferably LNP-formulated mRNA.

[0107] In one embodiment, the pharmaceutical production module comprises: i) a production device comprising a processing medium supply unit and a mixing unit, ii) a production device comprising a de novo DNA synthesis unit, iii) a production device comprising a purification unit, iv) a production device comprising a filtration unit, v) a production device comprising a processing medium supply unit and a mixing unit, vi) a production device according to the second aspect, optionally in combination with a DNA modification unit, vii) a production device comprising a purification unit, viii) a production device comprising a filtration unit, ix) a production device comprising a processing medium supply unit and a mixing unit, x) a production device comprising a bioreactor for RNA in vitro transcription, xi) a production device comprising a purification unit, xii) a production device comprising a filtration unit, xiii) a production device comprising a processing medium supply unit and a mixing unit, xiv) a production device comprising a filtration unit, preferably a tangential flow filtration unit, and xv) a production device comprising a filtration unit, preferably a sterile filter, which devices are coupled in a predetermined order. This module is preferably configured to produce formulated RNA, preferably formulated mRNA, more preferably LNP-formulated mRNA.

[0108] The pharmaceutical manufacturing module provides a very flexible and versatile platform. The pharmaceutical manufacturing module can be adapted for different purposes and applications. The pharmaceutical manufacturing module allows for modular manufacturing or production of pharmaceutical products. The pharmaceutical manufacturing module may be understood as an object suitable for modular manufacturing, meaning that the pharmaceutical manufacturing module can be adapted for different purposes. One object may be sufficient for a specific manufacturing method, or several objects may be connected, for example in the form of a manufacturing line or manufacturing chain, to carry out a specific manufacturing method. Such a modular design allows for easy handling, process improvements and / or maintenance. The techniques and operations used in the manufacturing module, as well as the selection of product contact materials, can be pre-evaluated before use, resulting in the production of high quality pharmaceutical products. As a result, the manufacturing module allows for a very robust and reproducible manufacturing of pharmaceutical products. Furthermore, the manufacturing module generally allows for the (fully or partially) automated manufacturing of pharmaceutical products.

[0109] In one embodiment, the processing medium supply unit comprises a mounting element configured to hold a processing medium supply container in the processing chamber. Typically, there are multiple mounting elements, each configured to hold a processing medium supply container, which is connected to a downstream unit or device, such as a DNA de novo synthesis unit, a DNA template generation unit, an RNA generation unit, or a formulation unit, for example, via a sterile tube. The downstream unit or device may be, for example, a purification unit, such as a reverse-phase HPLC column. The processing medium may be selected from the group consisting of synthetic DNA for amplification in a PCR reaction, a PCR component mixture, water, an in vitro translation buffer, nucleotides including UTP, GTP, ATP, and CTP, enzymes including RNA polymerase, buffers including wash buffers, reagent buffers (e.g., immobilization buffers), and elution buffers, HPLC buffers, formulation solutions including lipid solutions, and HPLC eluents.

[0110] In one embodiment, the mixing unit comprises a mixer. The mixing unit may also include a pump unit (e.g., a syringe pump, a pulseless pump, a peristaltic pump, etc.). The mixing unit may be used, for example, in combination with either a DNA template generation unit or an RNA generation unit, where the mixing unit is upstream of either the DNA template generation unit or the RNA generation unit and provides the respective master mix to the subsequent DNA template generation unit or the RNA generation unit. The mixing unit may also be used when a multivalent pharmaceutical product is produced, such as a multivalent RNA-based pharmaceutical product that includes at least two different RNA sequences. In the case of such a multivalent RNA-based pharmaceutical product, the mixing unit can be used to mix at least two different RNAs (i.e., at least two RNAs with different sequences) before these RNAs are formulated in the formulation unit. Alternatively, at least two different RNAs that have already been formulated may be mixed by the mixing unit to obtain a multivalent RNA-based pharmaceutical product.

[0111] The mixing unit may also be used when formulating nucleic acid products, particularly RNA, and may be referred to as a formulation unit in this case. Thus, the mixing unit may be used in formulations where lipid nanoparticles (LNPs), or liposome-encapsulated RNA, or RNA complexed with polycationic peptides or proteins (e.g., protamine or polymeric carriers, such as polyethylene glycol / peptide polymers as described in WO2012 / 013326) are produced. In the context of formulation, the mixing unit may include at least two pump units (e.g., one unit for pumping lipids, one unit for pumping RNA), and optionally a reactor (e.g., a T-piece connector) for complexation / formulation.

[0112] In one embodiment, the de novo DNA synthesis unit comprises a solid-phase synthesis unit for DNA, typically using the phosphoramidite method, to obtain DNA oligonucleotides of about 200 nucleotides in length, and corresponding phosphoramidite building blocks derived from protected 2'-deoxynucleosides. Preferably, in a fully automated manner, the phosphoramidite building blocks are successively linked together according to the sequence to be produced (sometimes referred to as an in silico designed sequence), and the oligonucleotide product is typically released from the solid phase into solution, then deprotected and recovered. The de novo DNA synthesis unit may further be configured to assemble (or link) at least two oligonucleotides obtained by the phosphoramidite method, if applicable, to obtain longer sequences. As mentioned above, a purification unit typically follows the de novo DNA synthesis unit. De novo DNA synthesis may also be based on an enzymatic process, for example terminal deoxynucleotidyl transferase (TdT) as described in WO2015159023. Furthermore, the de novo DNA synthesis unit may comprise a ligation unit for ligating the de novo synthesized DNA to a suitable DNA backbone.

[0113] In one embodiment, the DNA modification unit comprises suitable means for enzymatically or chemically modifying the amplified DNA. The enzymatic modification may for example be a digestion reaction with a restriction endonuclease to cleave the DNA at a specific position (for example to provide a poly(A) terminated DNA). The chemical modification may for example be binding to a tag or the like at the 5' and / or 3' end or chemical modification of the 5' and / or 3' end. Such a DNA modification unit may be operable in a continuous mode, in which the amplified PCR products obtained, for example, from a device according to the first aspect of the present application are fed to the DNA modification unit. When the DNA modification unit is configured for enzymatic DNA modification with a restriction endonuclease, it is preferred that the respective restriction endonuclease is immobilized (for example as described in WO2016174227). The DNA modification unit as defined above may in some embodiments be comprised in the device of the first aspect.

[0114] In one embodiment, the RNA production unit comprises a bioreactor for RNA in vitro transcription, preferably a bioreactor as disclosed in WO 2020 / 002598. In particular, a bioreactor as defined by claims 1 to 59 of WO 2020 / 002598 or as illustrated by Figures 1 to 14 of WO 2020 / 002598 can be used as an RNA production unit. It is particularly preferred for the bioreactor that in the bioreactor, the DNA used as template is immobilized on magnetic particles and that the bioreactor comprises a magnet for mixing the reaction solution containing the DNA immobilized on the magnetic particles.

[0115] In one embodiment, the purification unit comprises an HPLC unit, preferably a unit for performing RP-HPLC. Particularly preferred in this context is RP-HPLC using the methods disclosed in WO 2008 / 077592, preferably using a porous non-alkylated poly(styrene-divinylbenzene) reversed phase, the reversed phase being formed by beads or occurring as a polymerized block (e.g. monolithic). Alternatively or additionally, the purification unit may comprise an affinity chromatography unit, preferably an oligo dT purification unit for affinity purifying polyadenylated nucleic acids, in particular RNA, via oligo dT-functionalized matrices or beads or columns (e.g. as described in WO 2014152031 A1). Alternatively or additionally, the purification unit may comprise an anion exchange chromatography unit. Alternatively or additionally, the purification unit may comprise a unit for nucleic acid precipitation and a unit for purifying the precipitated nucleic acids (e.g. using TFF, or centrifugation, or filtration). Alternatively or additionally, the purification unit may comprise a unit for dsRNA removal, for example a unit comprising RNase III treatment or a unit comprising a step of dsRNA purification by cellulose. The purification unit may be used in particular to purify nucleic acids, preferably to purify template DNA obtained in the de novo DNA synthesis unit, DNA obtained in the device according to the first aspect, and / or RNA obtained in the RNA production unit. In a preferred embodiment, the purification unit, in particular the purification unit for purifying RNA, comprises an RP-HPLC unit, and an oligo dT purification unit, and optionally a unit for dsRNA removal.

[0116] In one embodiment, the filtration unit comprises a tangential flow filtration unit. Particularly preferred in this context is tangential flow filtration as described in WO 2016 / 193206, where TFF is used for diafiltration and / or concentration and / or purification of nucleic acid. The filtration unit can be used after the purification unit, in particular for filtering nucleic acid, preferably for filtering template DNA or RNA. It may also be preferred that the filtration unit comprises a sterile filter, preferably a sterile filter with a size of 0.22 μm, in particular for sterile filtering formulations that contain RNA, in particular mRNA, more particularly LNP-formulated mRNA.

[0117] In a preferred embodiment, the fill and finish unit comprises a filler and / or dispenser. The fill and finish unit is configured to fill the formulated drug product, in particular the formulated active pharmaceutical ingredient (in particular the formulated RNA, more particularly the LNP-formulated mRNA), into a suitable vial, such as a glass vial, in an amount corresponding to one or more doses of the formulated active pharmaceutical ingredient. Typically, the vial, such as a glass vial, is closed with a cap (e.g., an aluminum cap) under sterile conditions. In an embodiment, the fill and finish unit comprises a freezing unit for freezing the obtained drug product to at least -20°C, preferably at least -60°C or -80°C.

[0118] In a fourth aspect, the present invention is directed to a method for preparing a DNA product by capillary polymerase chain reaction. The method for preparing the DNA product may include, but is not necessarily in this order: - providing a first section, a second section, and a tube, the tube being wound at least from the first section to the second section and from the second section to the first section, the tube wound from section to section forming a helical stack of tube windings; - providing means for regulating the separate temperatures in each compartment; - directing the PCR solution through the tube, thereby directing the PCR solution through distinct temperatures in each compartment, and preparing a DNA product based on the PCR solution; Includes.

[0119] The method according to the invention for preparing DNA products allows for flexible adaptation to different conditions, requirements, and for example amounts of DNA. The method of the invention allows for the preparation of large amounts of DNA (for example, more than 2 mg per reaction, even up to g amounts). Furthermore, the preparation of DNA products can be scalable. The method of the invention for preparing DNA products allows for very rapid preparation of DNA products, and therefore also for rapid production of pharmaceutical products. Also, the modification and adaptation of the device for preparing DNA products to a specific DNA product can be very rapid. There may be no need for adjustment or calibration steps. There may be no need for washing steps or there may be less. For example, the turnover time for vaccine production can be less than about one week.

[0120] In one embodiment, the temperature in the first compartment is set for the denaturation step of the polymerase chain reaction and the temperature in the second compartment is set for the annealing step of the polymerase chain reaction and, optionally, for the extension step of the polymerase chain reaction.

[0121] In one embodiment, the temperature in the first compartment is in the range of about 85° C. to about 105° C., preferably about 98° C. In one embodiment, the temperature in the second compartment is in the range of about 45° C. to about 72° C., preferably about 60° C. to about 72° C.

[0122] In one embodiment, the method includes providing a third compartment. The tube is wound from at least the second compartment to the third compartment, and then back to the first compartment. The temperature in the third compartment is set for the extension step of PCR, preferably in the range of about 65°C to about 75°C, preferably about 72°C.

[0123] In one embodiment, the method for preparing the DNA product is carried out under GMP compliant conditions. In one embodiment, the means for regulating the separate temperatures within each compartment may be the provision of a separate heat transfer medium flow through each compartment via a heat transfer medium inlet and outlet for each compartment (to regulate the separate temperatures within each compartment).

[0124] In another embodiment, the means for regulating the separate temperatures within each compartment may be a heating unit (e.g., a heating cartridge) and optionally a cooling unit (e.g., a thermoelectric cooler) contained within each compartment (optionally connected to a ventilator).

[0125] In a fifth aspect, the present invention is directed to the use of a device for preparing a DNA product according to the first aspect for preparing a DNA product by capillary polymerase chain reaction, to the use of a pharmaceutical manufacturing device according to the second aspect for the production of a pharmaceutical product, and to the use of a manufacturing module according to the third aspect for the production of a pharmaceutical product.

[0126] The pharmaceutical may be formulated. The pharmaceutical is preferably an active pharmaceutical ingredient in the form of a biomolecule or any precursor or any intermediate thereof, the biomolecule is preferably a nucleic acid, more preferably DNA. The manufacturing is preferably GMP-compliant.

[0127] In one embodiment, said use is automated. This means that the use or operation of the device for preparing a DNA product by capillary polymerase chain reaction according to the first aspect, the pharmaceutical product manufacturing device according to the second aspect, or the manufacturing module according to the third aspect can be fully or partially automated. This can allow for more reliable, faster and / or more cost-effective production of pharmaceutical products.

[0128] It is noted that the above aspects of the invention apply to the device for preparing a DNA product by capillary polymerase chain reaction, the device for producing a medicinal product, the module for producing a medicinal product, the method for preparing a DNA product by capillary polymerase chain reaction, the use of the device for preparing a DNA product by polymerase chain reaction and the use of the device for the production of a medicinal product as defined in the independent claims, which have similar and / or identical preferred embodiments, in particular as defined in the dependent claims.Furthermore, it is understood that a preferred embodiment of the invention may also be any combination of the dependent claims with the respective independent claims.

[0129] These and other aspects of the invention will be apparent from and elucidated with reference to the embodiments described hereinafter. [Brief description of the drawings]

[0130] The figures shown below are merely examples and further illustrate the invention and should not be construed as limiting the invention to those figures. [Figure 1] FIG. 1 shows a top view of a device according to the invention for preparing DNA products by capillary polymerase chain reaction. [Diagram 2] FIG. 1 shows a side view of a device according to the invention for preparing DNA products by capillary polymerase chain reaction. [Diagram 3] A 3D side view of the rolled tube is shown. [Figure 4] Shows a 3D side view of a tube wrapped around several stands. [Diagram 5] 3D diagram of a pharmaceutical manufacturing device. [Figure 6a] 3D diagram of a box containing a device for preparing a DNA product, which is inserted into a pharmaceutical manufacturing device. [Figure 6b] 3D diagram of a box containing a device for preparing a DNA product, which is inserted into a pharmaceutical manufacturing device. [Figure 7]1 shows a schematic and exemplary representation of a pharmaceutical production module according to the invention; [Figure 8] 1 shows a 3D side view of a device according to the invention. [Figure 9] FIG. 2 shows a 3D side view of the top of a device according to the invention. [Figure 10] 3D side view of a box containing a device to be inserted into a manufacturing device. [Figure 11] A close-up 3D view of the manufactured device is shown with the box containing the device inserted into the manufactured device. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0131] FIG. 1 shows a top view of a device 1 according to the invention for preparing a DNA product by capillary polymerase chain reaction. FIG. 2 shows a side view of a device 1 according to the invention for preparing a DNA product by capillary polymerase chain reaction. FIG. 8 shows a 3D side view of the device according to the invention. In FIG. 2, the device 1 for preparing a DNA product is mounted in a pharmaceutical manufacturing device 10 according to the invention, which will be explained in more detail with reference to FIG. 5. The DNA product can be DNA to be used as an API or a DNA template for enzymatic RNA in vitro transcription (RNA is the API). Polymerase chain reaction (PCR) is a technique to synthetically amplify DNA over several orders of magnitude to generate thousands to millions of copies of a specific DNA sequence. Capillary polymerase chain reaction can be understood as a PCR carried out in a capillary or tube 6. The present device 1 for preparing a DNA product allows flexible and rapid preparation of large amounts of DNA product. The device 1 for preparing a DNA product can be used for flexible and rapid vaccine production, in particular for DNA vaccine production or for the production of DNA templates for enzymatic RNA in vitro transcription. The device 1 may in particular be used to prepare DNA templates for mRNA-based vaccines, for example during infectious disease epidemics and pandemics.

[0132] As shown in Figures 1 and 8, the device 1 for preparing DNA products 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 that can range from about 0.5 mm to about 50 mm, preferably from about 0.5 mm to about 1.5 mm, more preferably from 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. The tube 6 then returns to the first compartment 7. The PCR solution can be filled into the tube 6 and thereby guided through the three compartments. As shown in Figures 2 and 8, the compartments can be understood as independent tanks that are filled, for example, by a heat medium (see Figure 2) or a heat solid (see Figure 8). Here, the vessel is at least partially, and preferably completely, surrounded by insulation, which may be a bulkhead 14 (see FIG. 1) or an internal compartment wall 30 (see FIG. 8) (optionally combined with bulkhead 14).

[0133] The tube 6 is wound to form a helical stack of tube turns, which can be understood as a helix, whorl or coil of turns when viewed in side view. Figure 3 shows a 3D side view of the wound tube 6. There can be several layers of turns (e.g. 30). The helical extension or stacking direction of the tube 6 extends in the z direction, which is essentially perpendicular to the x,y plane formed by the bottom of the device 1 for preparing DNA products, or the plane formed by the floor.

[0134] 1 and 8, the tube 6 is not only wound from the first section 7 to the second section 8 and from the second section 8 to the third section 15, but also from the third section 15 to the first section 7, from the first section 7 to the second section 8 and from the second section 8 to the third section 15, etc. Each tube winding represents one PCR cycle.

[0135] The tube 6 extends in a wavy manner in the three compartments, which can be understood by the fact that, when viewed in top view, the tube 6 has a wave or snake shape inside all three compartments. The corrugation of the tube 6 extends in the x- and y-directions of the plane formed by the bottom or the plane formed by the floor of the device 1 for preparing DNA products. The corrugation form of the tube 6 defines the residence time of the PCR liquid in each compartment. The corrugation direction of the tube 6 is essentially perpendicular to the stacking direction of the tubes 6.

[0136] As shown in Fig. 1, the device for preparing a DNA product 1 further comprises a PCR liquid inlet 20 for a PCR mixture, which is connectable to an external pumping unit or pump (37) for pumping the PCR liquid through the tube 6. The device for preparing a DNA product 1 further comprises a DNA product outlet 25 for the PCR liquid product. Alternatively, the PCR device may comprise an internal pumping 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 in the first compartment, while the DNA product outlet 25 may be located in the third compartment.

[0137] As shown in Figures 1 and 2, the device 1 for preparing DNA products in this embodiment further comprises a heat medium inlet 12 and a heat medium outlet 13 in each of the three compartments. As mentioned above, the device 1 may comprise other means for adjusting the temperature in each compartment. When a heat medium is used, the heat medium can enter each of the three compartments by the respective heat medium inlet 12 and can exit each of the three compartments by the respective heat medium outlet 13. This can result in a flow of the heat medium through each compartment. The heat medium outlet 13 is preferably located at the top to facilitate the removal of air bubbles.

[0138] The heat 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 temperature zones correspond to the PCR temperature profile for denaturation, annealing, and extension. The PCR liquid is separated from the heat medium in the compartment by a tube wall. The tube wall allows a temperature exchange between the PCR liquid and the interior of the compartment, e.g., the heat medium, which can be used to heat or cool the PCR liquid, e.g., by the heat medium.

[0139] The heat medium inlet 12 and the heat medium outlet 13 are arranged in each compartment in this embodiment to provide a heat medium flow through the compartment in essentially the opposite direction relative to the guiding direction of the PCR fluid in the tube 6. As shown in Figure 1, this can be understood to mean that in the case of a guiding direction of the PCR fluid from the top left to the bottom right, the heat medium inlet 12 and the heat medium outlet 13 are arranged to provide a heat medium flow in the opposite direction from the bottom right to the top left (see the arrow in the first compartment 7). The counter-flow of the heat medium and the PCR fluid improves the temperature exchange between the heat medium and the PCR fluid.

[0140] As shown in FIG. 8, the device 1 in this embodiment comprises an isolating section 28, an outer wall 29 and an inner compartment wall 30 that corrugates and surrounds the tubes in each of the compartments 7, 8 and 15. Each compartment comprises at least one heating unit 31 (here shown as a heating cartridge) as well as a cooling unit, here a heat pipe 32, that can be used for cooling if necessary. In this embodiment, the space in each compartment enclosed by the inner compartment wall 30 (this space surrounding the tubes 6) is filled with a thermal solid (such as, for example, aluminum pellets), optionally combined with a liquid or gel, which is heated by the at least one heating unit 31 as well as cooled, if necessary, by a cooling unit (comprising a heat pipe 32) to provide a specific temperature in each compartment. The space between the compartments and / or the surrounding outer wall 29, i.e. the space not surrounding the tubes 6, can be filled with a synthetic material (for example synthetic balls or granules). FIG. 9 shows a top view of the device 1 shown in FIG. 8, illustrating the isolating section 28 and the outer wall section 29, and also illustrating the upper part of the heating unit 31 and 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 carry away heat. Also, in this embodiment, three different temperature zones are provided in the three compartments to prepare DNA products by thermal cycling. The three temperature zones correspond to the temperature profile of PCR for denaturation, annealing, and extension. The PCR liquid is separated from the thermal solids in the compartments by the tube walls.

[0141] As shown in Fig. 1 and Fig. 8, the device 1 for preparing DNA products further comprises partitions 14 between the compartments to separate them from each other (in Fig. 8 the partitions 14 are only partially present since the internal compartment wall 30 already separates the compartments from each other). One partition 14 is placed between the first compartment 7 and the second camber, another partition 14 is placed between the second camber compartment and the third camber, and a further partition 14 is placed between the third camber compartment and the first camber. As a result, adjacent compartments are insulated from each other and therefore separate temperatures can be established and maintained in adjacent compartments. The partitions 14 can be plate-shaped elements. The partitions 14 between the compartments can be implemented by several walls or by only one (e.g. T-shaped or star-shaped) wall. The partitions 14 comprise through holes for the tubes 6 to extend from one compartment to the next.

[0142] As shown in Figures 1 and 9, the device 1 for preparing a DNA product further comprises a sensor 21 arranged in each compartment, here for example a temperature sensor 21. Preferably, several of these temperature sensors are present in each compartment.

[0143] The device 1 for preparing DNA products further comprises a stand 17 for holding the tubes 6. Figure 4 shows a 3D side view of the tubes 6 wound around several stands 17. The stands 17 are also shown in Figures 1 and 8. A stand 17 is placed in each compartment to hold and guide the tubes 6. The stand 17 provides a fixing point and a deflection point for the corrugation of the tubes 6. The stand 17 may therefore comprise a slit or the like for attaching the tubes 6 (wound) to the stand 17. The tubes 6 extend partly around the outer dimension of the stand 17. The stand 17 here has a T- or L-shape when seen in top view, but other shapes are also possible, for example a cylindrical shape. The stand 17 may be exchangeable and repositionable within the compartment. The tubes 6 are here the same throughout the device 1 for preparing DNA products and may be exchangeable.

[0144] Figure 5 shows a 3D view of a pharmaceutical production device 10. Figure 11 shows a cut-out view thereof. The pharmaceutical production device 10 comprises a housing 5, a processing chamber 2, a technological chamber 3 and a device 1 for preparing a DNA product by capillary polymerase chain reaction as described above. A 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 pharmaceutical production device 10 can be operated under conditions compliant with GMP (Guidelines for Good Manufacturing Practice).

[0145] The pharmaceutical production device 10 can be understood as a closed and sealed housing 5, containing different chambers for different purposes. The processing chamber 2 is suitable and used for producing pharmaceutical products. The processing chamber 2 is here a grade A room. The processing chamber 2 is sealed against the technical chamber 3. The technical chamber 3 is suitable and used for housing devices such as pumps, motors, mixers, processors, etc. 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 devices thereon.

[0146] The technological chamber 3 and the processing chamber 2 are arranged in such a way that the gas flow in the technological chamber 3 is parallel to, but in the opposite direction to, the gas flow in the processing chamber 2. The gas of the gas flow is here clean air. There is a gas inlet duct 9a and a gas outlet duct 9b.

[0147] At least one control unit controls the gas flow through the processing chamber 2 to provide a gas shower falling in the direction of gravity (downward) and / or a positive pressure in the processing chamber 2. The control unit can be a valve or a processor. The gas shower can be a laminar flow. The gas flow can be provided by a flow unit, which can be located inside or outside the housing 5 and can be in communication with the control unit. The flow unit can be a pump, an HVAC system, etc. The positive pressure is an overpressure relative to the environment outside the housing 5. The control unit regulates the different pressures of the gas flow while the flow unit provides a variable amount of gas.

[0148] As shown in Fig. 5, the separation element 4 comprises at least an attachment 16 with an opening towards the processing chamber 2 and an extension (extended processing chamber) in the direction of the technology chamber 3. The attachment 16 is used to accommodate an apparatus such as, for example, the above-mentioned device 1 for preparing a DNA product (sometimes called a "PCR reactor"). This allows to increase the available space in the processing chamber 2. The device 1 for preparing a DNA product is accommodated here in a box 26, which is inserted into the attachment 16 as a single element. The box 26 may be replaceable.

[0149] As also shown in Fig. 5, when the device 1 for preparing a DNA product is inserted into the attachment 16, the front of the device 1 for preparing a DNA product, specifically the front of the box 26, ends essentially flush with the opening of the attachment 16 into the processing chamber 2. As a result, the gas flow, specifically the gas shower, is seamless and undisturbed or less disturbed and / or the precipitation surface is reduced. The attachment 16 and here its front is closed by a cover. The cover may be part of the box 26 of the device 1 for preparing a DNA product.

[0150] 5, the pharmaceutical manufacturing device 10 further comprises a fluid collection tray 11 for collecting leaked fluids. The fluid collection tray 11 may be located at the bottom of the housing 5 and / or in the passage from the processing chamber 2 to the technical chamber 3. The fluid collection tray 11 may comprise a leak sensor 21.

[0151] Figure 11 shows the rear side of the manufacturing device 10 into which the device 1 is inserted. At the rear side facing the technological chamber 3, there is a ventilator 34. Furthermore, there is an air inlet 35, together with an electronic plug 36 for connecting the device 1 to the technological medium in the technological chamber, in particular to the electrical power. This embodiment relates in particular to devices in which a thermal solid is used to regulate the temperature of the compartments (for example as illustrated in Figures 8, 9 and 10).

[0152] 6a and 6b as well as 10 show a 3D view of a box 26 containing a device 1 for preparing a DNA product to be inserted into the device 10 for the manufacture of a pharmaceutical product. The box 26 is closed by a cover. The cover comprises a handle 22 for handling the device 1 for preparing a DNA product in and / or out of the attachment 16. The box 26 comprises a heat pipe 32 (see fig. 10) connected to a thermoelectric cooler 33 and a ventilator 34, as well as inlets 12 and outlets 13 (see fig. 6b) for the heat medium for each compartment of the device 1 for preparing a DNA product or a heating cartridge 31. The inlet 23 for the PCR liquid and / or the outlet for the DNA product 24 are arranged at the front, here on the cover of the box 26 of the device 1 for preparing a DNA product. The attachment 16 (see fig. 5) and the box 26 comprise corresponding parts of an exchange rail system 27 that guides and facilitates the exchange of the device in the attachment 16, for example to exchange the device 1 for preparing a DNA product with another device. As shown in Figures 6b and 10, a sensor 21, e.g. a leak sensor 21 or a temperature sensor 21, is arranged in a box 26. The box 26 further comprises a connection, e.g. an electrical connection, which is connected to the processing chamber 2. In the embodiment according to Figure 10, the sensor is preferably a temperature sensor, since the embodiment of Figure 10 (where a thermal solid is used as heating means) typically does not include a leak sensor (as no thermal medium is used in the embodiment shown in Figure 10).

[0153] An external pumping unit or pump (37 in FIG. 7) for pumping PCR fluids through tubes 6 of device 1 for preparing DNA products may be located outside device 1, for example within a processing chamber of pharmaceutical manufacturing device 10.

[0154] 7 shows diagrammatically and exemplarily a manufacturing module 100 for a pharmaceutical product according to the invention, which here comprises four manufacturing devices 10. The manufacturing devices are very flexible, as several manufacturing devices can be adapted for several manufacturing processes, for example forming a manufacturing chain.

[0155] The four manufacturing devices 10 are connected to each other by a connection unit 18 arranged on the outer wall of the housing 5. The connection unit 18 allows for fluid communication and connection, preferably also mechanical connection, and data communication and connection of the manufacturing devices 10. The four manufacturing devices 10 are exemplarily shown here as (from the left) a first device with a process medium supply unit and a mixing unit, a second device with a manufacturing device 10 of a pharmaceutical product as described above (device 1 for preparing a DNA product by capillary polymerase chain reaction), a third device with a purification unit, and a fourth device with a filtration unit, which comprises two filtration units, namely a tangential flow filtration unit and a sterile filter. This exemplary module can be used for DNA production and DNA amplification, and purification, respectively. The exemplary module used for DNA production / amplification may be connected to a further manufacturing device (e.g. a manufacturing device with an RNA bioreactor as described in WO2020002598) that transcribes the obtained DNA into RNA.

[0156] The manufacturing module 100 comprises a mounting element 19 arranged on an outer wall of the housing 5 of the manufacturing device 10 outside the housing 5. The mounting element 19 is configured to hold a medium supply (not shown), for example in the form of a medium reservoir.

[0157] definition For clarity and ease of reading, the following definitions are provided. Any technical features mentioned with respect to these definitions can be read for each and every embodiment of the present invention. Further definitions and explanations may be specifically provided in the context of these embodiments.

[0158] As used in this specification and claims, the singular forms "a" and "an" include the corresponding plural forms unless the context clearly dictates otherwise. The term "about" in the context of the present invention indicates an interval of accuracy that a person skilled in the art would understand to still ensure the technical effect of the feature in question. This term typically indicates a deviation of ±10%, preferably ±5% from the indicated numerical value.

[0159] It should 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". Hereinafter, when a group is defined as comprising at least a certain number of embodiments, this also means to encompass the group preferably consisting of only these embodiments.

[0160] The term "pharmaceutical product" as used herein relates to an active pharmaceutical ingredient or any precursor or any intermediate thereof. Thus, a "pharmaceutical product" is specifically an active pharmaceutical ingredient, which is used in a medicine and administered to a human or animal subject for the treatment or prevention of a disease, i.e. of clinical grade, in particular with respect to parameters such as purity, integrity, etc. Such pharmaceutical products are typically produced in vitro in a synthetic process, which includes precursors and intermediates in the process of production. In 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 be used in particular as a vaccine or as a DNA template for RNA in vitro transcription. For example, when producing mRNA as the active pharmaceutical ingredient, the first step may be the production of a template DNA (also referred to herein as "DNA template"), which corresponds to a precursor of the mRNA, since it serves as a template in an in vitro transcription reaction in producing the RNA. The intermediate in DNA production may be raw DNA obtained from a PCR reaction, as such DNA does not represent the final active pharmaceutical ingredient, but requires inter alia purification steps to be performed in order to provide the required clinical grade product.

[0161] The term "DNA" is the customary abbreviation for deoxyribonucleic acid. It is a nucleic acid molecule, i.e., a polymer made up of nucleotide monomers. These nucleotides are usually deoxyadenosine monophosphate, deoxythymidine monophosphate, deoxyguanosine monophosphate, and deoxycytidine monophosphate monomers or their analogs, which themselves consist of a sugar moiety (deoxyribose), a base moiety, and a phosphate moiety, polymerized with a characteristic backbone structure. The backbone structure is typically formed by a phosphodiester bond between the sugar moiety, i.e., deoxyribose, of a first nucleotide and the 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 can 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.

[0162] The term "RNA" is the customary abbreviation for ribonucleic acid. It is a nucleic acid molecule, i.e. a polymer made up of nucleotide monomers. These nucleotides are usually adenosine monophosphate (AMP), uridine monophosphate (UMP), guanosine monophosphate (GMP) and cytidine monophosphate (CMP) monomers or their analogs, which are linked together along a so-called backbone. The backbone is formed by a phosphodiester bond between a first sugar, i.e. a ribose, and the 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, for example, by transcription of a DNA sequence in a cell. In eukaryotic cells, the transcription typically takes place in the nucleus or mitochondria. In vivo, transcription of DNA usually results in a so-called immature RNA, which must be processed into a so-called messenger RNA (usually abbreviated to mRNA). For example, the processing of immature RNA in eukaryotes includes a variety of different post-transcriptional modifications, such as splicing, 5' cap formation, polyadenylation, and export from the nucleus or mitochondria. The whole process is also called RNA maturation. Mature messenger RNA usually provides a nucleotide sequence that can be translated into the amino acid sequence of a specific peptide or protein. Typically, mature mRNA includes a 5' cap, an optional 5' UTR, a coding sequence, an optional 3' UTR, and a poly(A) sequence. When the RNA molecule is of synthetic origin, as in the present invention, it means that the RNA molecule is produced in vivo, i.e., not in a cell or purified from a cell, but by an in vitro method. An example of a suitable in vitro method is in vitro transcription. In addition to messenger RNA, there are several non-coding types of RNA that may be involved in transcription and / or translation regulation, immune stimulation, and these can 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, immunostimulatory RNA, transfer RNA (tRNA), ribosomal RNA (rRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), microRNA (miRNA), and Piwi-interacting RNA (piRNA).

[0163] The term "RNA in vitro transcription" refers to a process in which RNA is synthesized in a cell-free system. RNA can be obtained by DNA-dependent RNA in vitro transcription of a suitable DNA template, which can 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. Specific examples of DNA-dependent RNA polymerases are T7 RNA polymerase, T3 RNA polymerase, SP6 RNA polymerase, or Syn5 RNA polymerase.

[0164] Reagents used for RNA in vitro transcription typically include: a DNA template (linearized DNA or linear PCR product) with a promoter sequence that has high binding affinity for the respective RNA polymerase, such as a bacteriophage-encoded RNA polymerase (T7, T3, SP6, or Syn5); ribonucleotide triphosphates (NTPs) of the four bases (adenine, cytosine, guanine, and uracil); optionally, a cap analog (e.g., m7G(5')ppp(5')G (m7G) or a cap analog derivable from the structures disclosed in claims 1-5 of WO2017 / 053297, or any cap structure derivable from the structures defined in claim 1 or claim 21 of WO2018075827); optionally, further modified nucleotides, as defined herein; a DNA-dependent RNA polymerase (e.g., T7, T3, SP6, or Syn5) capable of binding to the promoter sequence in the DNA template. RNA polymerase; optionally an RNase inhibitor to inactivate potentially contaminating ribonucleases (RNases); optionally a pyrophosphatase (an inhibitor of RNA synthesis) to degrade pyrophosphate; MgCl2 to provide Mg2+ ions as a cofactor for the polymerase; a buffer (TRIS or HEPES) to maintain a suitable pH value, which may also contain an antioxidant (e.g., DTT), and / or an optimal concentration of a polyamine such as spermidine, for example a buffer system comprising citrate and / or betaine as disclosed in WO 2017 / 109161.

[0165] The nucleotide mixture used for RNA in vitro transcription may further contain modified nucleotides as defined herein. In this context, preferred modified nucleotides include pseudouridine (ψ), N1-methylpseudouridine (m1ψ), 5-methylcytosine, and 5-methoxyuridine. The nucleotide mixture used for RNA in vitro transcription reaction (i.e., the fraction of each nucleotide in the mixture) may be optimized for a given RNA sequence, preferably as described in WO2015188933.

[0166] An "RNA in vitro transcription (IVT) master mix" may include components necessary for carrying out an RNA in vitro transcription reaction as defined above. Thus, the IVT master mix may include at least one component selected from a nucleotide mixture, a cap analog, a DNA-dependent RNA polymerase, an RNase inhibitor, a pyrophosphatase, MgCl2, a buffer, an antioxidant, betaine, and citrate.

[0167] As used herein, the term "template DNA" (or "DNA template") typically refers to a DNA molecule that comprises a nucleic acid sequence that codes for an RNA sequence to be transcribed in vitro. The template DNA is used as a template for RNA in vitro transcription to produce the RNA encoded by the template DNA. Thus, the template DNA contains all the elements necessary for RNA in vitro transcription, in particular a promoter element 5' to the DNA sequence coding for the target RNA sequence for binding of a DNA-dependent RNA polymerase, such as, for example, T3, T7, and SP6 RNA polymerase. Furthermore, the template DNA may contain primer binding sites 5' and / or 3' to the DNA sequence coding for the target RNA sequence, and the presence of the DNA sequence coding for the target RNA sequence may be determined, for example, by PCR or DNA sequencing.

[0168] "Polymerase chain reaction" or "PCR" is a molecular biology technique used to amplify a piece of DNA by several orders of magnitude, generating thousands to millions of copies of a specific DNA sequence. The method relies on thermal cycling, which consists of repeated cycles of heating and cooling the reaction for DNA melting and enzymatic replication of the DNA. The combination of primers (short DNA fragments) containing sequences complementary to the target sequence and a thermostable DNA polymerase such as Taq polymerase allows for selective and repetitive amplification. As PCR proceeds, the DNA produced is itself used as a template for replication, initiating a chain reaction in which the DNA template is exponentially amplified. DNA polymerase enzymatically assembles new DNA strands from nucleotides, the building blocks of DNA, using single-stranded DNA as the PCR template and DNA oligonucleotides (also called DNA primers) required to initiate DNA synthesis. Most PCR methods use thermal cycling, i.e., alternating heating and cooling of 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 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, which selectively amplifies the target DNA. The selectivity of PCR is achieved by using primers complementary to the DNA region to be amplified under specific thermal cycling conditions.

[0169] A "PCR master mix" or a "PCR solution" may contain components necessary for carrying out PCR as defined above. Thus, a PCR master mix may contain at least one component selected from a nucleotide mixture, a DNA polymerase, a synthetic DNA as an (initial) template, and a buffer solution.

[0170] The term "lipid nanoparticle" or "LNP" refers to a pharmaceutical formulation, specifically a formulation of a nucleic acid. In the context of the present invention, the term "LNP" is not limited to a particular form and includes any form that is generated when cationic lipids and optionally one or more additional lipids are combined, for example, in an aqueous environment and / or in the presence of nucleic acid. For example, liposomes, lipid complexes, lipoplexes, etc. are within the scope of lipid nanoparticles (LNPs). LNPs typically include cationic lipids and one or more excipients selected from neutral lipids, charged lipids, steroids, and polymer-conjugated lipids (e.g., PEGylated lipids). Nucleic acids can be encapsulated within the lipid portion of the LNP, or within the aqueous space enveloped by some or all of the lipid portion of the LNP. In one embodiment, the LNPs consist 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; and 0.5-15% PEG lipid.

[0171] The term "processing medium" refers to components that are directly and physically involved in any reaction or any processing step required to produce a pharmaceutical product. Thus, since production takes place in the processing chamber, the "processing medium" will be present in the processing chamber when the device is used for production. Thus, the processing medium may specifically be any starting material (e.g., nucleotides, etc.), any catalytic material (e.g., enzymes, etc.), and any buffer (e.g., reaction buffer or purification buffer, etc.). The processing medium is typically provided in a processing medium supply vessel.

[0172] The term "technical medium" refers to a component that is not directly involved in any reaction or any process step required to produce a pharmaceutical product. Rather, the technical medium is indirectly involved, for example as a power cable, which supplies power to a unit or device that processes the process medium and which would be located in the technical chamber. A technical medium source would be, for example, the power or power supply provided by the power cable.

[0173] It should be noted that the embodiments of the present invention are described with reference to separate subject matter. In particular, some embodiments are described with reference to method type claims, while other embodiments are described with reference to device type claims. However, those skilled in the art will understand from this specification that, unless otherwise specified, any combination of features belonging to one type of subject matter, as well as any combination between features related to different subjects, is also considered to be disclosed in this application. However, all features can be combined to provide a synergistic effect that is more than the simple sum of the features.

[0174] Where a group is defined as comprising at least a certain number of embodiments, this is also meant to encompass the group preferably consisting of only those embodiments. While the invention has been illustrated and described in detail in the drawings and specification, such illustration and description are to be considered as 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 the claimed invention, from a study of the drawings, the disclosure, and the dependent claims.

[0175] In the claims, the word "comprising" does not exclude other elements or steps, and the indefinite articles "a" or "an" do not exclude a plurality. The mere fact that certain measures are recited 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 interpreted as limiting the scope.

[0176] A set of embodiments of the present application relate to the following: 1. A device (1) for preparing DNA products by capillary polymerase chain reaction (PCR), comprising: A tube (6) for conducting the PCR solution; A first compartment (7); At least a second compartment (8), A tube (6) is wound from at least a first section (7) to a second section (8); A device (1), wherein a first compartment (7) and a second compartment (8) comprise a heat medium inlet (12) and a heat medium outlet (13) for providing separate heat medium flows through the first compartment (7) and the second compartment (8), respectively, to regulate separate temperatures within the first compartment (7) and the second compartment (8) to prepare DNA products based on PCR solutions.

[0177] 2. A device (1) for preparing a DNA product as described in embodiment 1, wherein a partition (14) is disposed between the compartments and isolates the compartments from each other. 3. A device (1) for preparing a DNA product according to the previous embodiment, wherein the septum (14) comprises a through hole for a tube (6) to extend from one compartment to the next.

[0178] 4. A device (1) for preparing a DNA product according to one of the previous embodiments, wherein a heat medium inlet (12) and a heat medium outlet (13) are arranged in at least one compartment, preferably in each compartment, so as to provide a heat medium flow through the compartment in an essentially opposite direction relative to the guiding direction of the PCR liquid in the tube (6).

[0179] 5. A device (1) for preparing a DNA product according to one of the previous embodiments, wherein a heat transfer medium inlet (12) is arranged in a lower position in each of the compartments and a heat transfer medium outlet (13) is arranged in an upper position in each of the compartments.

[0180] 6. The device (1) for preparing a DNA product according to one of the previous 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).

[0181] 7. The device (1) for preparing a DNA product according to the previous 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).

[0182] 8. A device (1) for preparing a DNA product according to one of the previous embodiments, in which the tube (6) wound from compartment to compartment forms a helical stack of tube windings.

[0183] 9. A device (1) for preparing a DNA product according to one of the previous embodiments, wherein the tube (6) extends in a wavy manner within at least one compartment, preferably within each of the compartments.

[0184] 10. A device (1) for preparing a DNA product according to embodiments 8 and 9, wherein the stacking direction of the spiral stack of tube windings is different from the corrugation direction of the corrugated tube (6) and is preferably essentially perpendicular to the corrugation direction of the corrugated tube.

[0185] 11. A device (1) for preparing a DNA product according to one of the previous embodiments, in which each compartment forms a reservoir which is filled by a heat transfer medium. 12. A device (1) for preparing a DNA product according to one of the previous embodiments, further comprising at least a stand (17) arranged in one of the compartments to provide a fixing point for the tube (6) and / or a deflection point for the corrugation of the tube (6).

[0186] 13. A device (1) for preparing a DNA product according to the previous embodiment, wherein the tube (6) extends partially, completely or more than twice around the outer dimension of the stand (17).

[0187] 14. A device (1) for preparing a DNA product according to any one of embodiments 12 to 13, wherein the stand (17) is replaceable and / or repositionable within the compartment. 15. The device (1) for preparing a DNA product according to one of the previous embodiments, wherein the tube (6) is provided with a PCR liquid inlet (20), which is connectable to a pump unit for pumping PCR liquid through the tube (6).

[0188] 16. A device (1) for preparing a DNA product according to the previous embodiment, wherein the pump unit is arranged outside the compartment, preferably outside the outer wall (5) surrounding the compartment.

[0189] 17. The device (1) for preparing a DNA product according to embodiment 15 or 16, wherein the pump unit is configured to provide a PCR solution at a flow rate in the range of about 0.1 to about 10 mL / min, preferably about 0.2 to about 3 mL / min.

[0190] 18. The device (1) for preparing a DNA product according to one of the previous embodiments, wherein the tube (6) is provided with a DNA product outlet (25) for continuous preparation of the DNA product. 19. A device (1) for preparing a DNA product according to any one of embodiments 1 to 17, wherein the tube (6) is provided with a closable DNA product outlet (25) for discontinuous preparation of the DNA product.

[0191] 20. The device (1) for preparing a DNA product according to one of the previous embodiments, further comprising at least a sensor (21) arranged in at least one of the compartments, the sensor (21) being configured to detect the temperature, flow rate or leakage of the compartment.

[0192] 21. The device (1) for preparing a DNA product according to one of the previous embodiments, wherein the tube (6) has a diameter in the range of about 0.5 to about 1.5 mm, preferably about 0.75 to about 1.25 mm.

[0193] 22. The device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) has a length between the tube inlet and the tube outlet in the range of about 10 to about 200 m, preferably about 25 to about 50 m.

[0194] 23. A device (1) for preparing a DNA product according to one of the preceding embodiments, wherein the tube (6) is sized to accommodate a PCR solution in the range of about 10 mL to about 1 L, preferably about 20 mL to about 250 mL.

[0195] 24. A device (1) for preparing a DNA product according to one of the previous embodiments, wherein the spiral stack of tube turns comprises from about 10 to about 50 turns, preferably from about 15 to about 40 turns.

[0196] 25. A 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 or more, preferably in the range of about 1 mg to about 30 mg, more preferably in the range of about 15 mg to about 25 mg of DNA product per reaction.

[0197] 26. The 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 a range of about 50 μg to about 150 μg of DNA product per mL of PCR solution, preferably about 75 μg to about 125 μg of DNA product per mL of PCR solution.

[0198] 27. A device (10) for the production of a pharmaceutical product, comprising a processing chamber (2), a technological chamber (3) and a device (1) for preparing a DNA product by capillary polymerase chain reaction according to one of the previous embodiments.

[0199] 28. The pharmaceutical manufacturing device (10) according to the previous embodiment, further comprising a control unit configured to control the gas flow through the processing chamber (2) as a gas shower. 29. A pharmaceutical manufacturing device (10) according to one of embodiments 27 to 28, wherein at least the processing chamber (2), preferably the manufacturing device (10), is operable in accordance with the requirements of Annex 1 of the EU guidelines on Good Manufacturing Practice (GMP).

[0200] 30. A pharmaceutical manufacturing device (10) according to any one of embodiments 27 to 29, further comprising a separation element (4) separating the processing chamber (2) from the technical chamber (3), the separation element (4) comprising an attachment (16) protruding in the direction of the technical chamber (3) having an opening facing the processing chamber (2), and a device (1) for preparing a DNA product can be inserted into the attachment (16).

[0201] 31. A pharmaceutical manufacturing device (10) as described in the previous embodiment, wherein when the device (1) for preparing a DNA product is inserted into the attachment (16), the front surface of the device (1) for preparing a DNA product terminates essentially flush with the opening of the attachment (16) into the processing chamber (2).

[0202] 32. A pharmaceutical manufacturing device (10) described in one of embodiments 27 to 31, in which at least a handle (22) for handling the device (1) for preparing a DNA product in and / or outside the attachment part (16) is arranged on the front side of the device (1) for preparing a DNA product.

[0203] 33. A pharmaceutical manufacturing device (10) according to one of embodiments 27 to 32, in which the inlet (23) for the PCR liquid and / or the outlet (24) for the DNA product are arranged at the front of the device (1) for preparing the DNA product.

[0204] 34. A pharmaceutical manufacturing device (10) described in one of embodiments 27 to 33, wherein the heat medium inlet and / or the heat medium outlet are located on the rear plate of the device (1) for preparing a DNA product, opposite the front plate of the device (1) for preparing a DNA product.

[0205] 35. A pharmaceutical manufacturing device (10) according to one of embodiments 27 to 34, further comprising a leak sensor (21) arranged in an outer attachment (16) of the device (1) for preparing a DNA product.

[0206] 36. A pharmaceutical manufacturing device (10) according to one of embodiments 27 to 35, wherein the attachment part (16) is provided with an exchange rail (27) for guiding the exchange of the device (1) for preparing a DNA product with another device (1) for preparing a DNA product.

[0207] 37. A pharmaceutical manufacturing device (10) according to one of embodiments 27 to 36, in which a pump unit for pumping PCR liquid through the tube (6) of the device (1) for preparing a DNA product is arranged outside the casing of the pharmaceutical manufacturing device (10).

[0208] 38. A pharmaceutical manufacturing device (10) according to one of embodiments 27 to 37, further comprising a coupling unit (18) arranged in a casing of the pharmaceutical manufacturing device (10) and configured to couple the pharmaceutical manufacturing device (10) to another manufacturing device (10).

[0209] 39. A pharmaceutical manufacturing module (100) comprising a pharmaceutical manufacturing device (10) described in one of embodiments 27 to 38, and at least one of the group consisting of a manufacturing device (10) having a processing medium supply unit and a mixing unit, a manufacturing device (10) having a purification unit, and a manufacturing device (10) having a filtration unit.

[0210] 40. A method for preparing a DNA product by capillary polymerase chain reaction (PCR), comprising: - providing a first section (7), a second section (8) and a tube (6), the tube (6) being wound at least from the first section (7) to the second section (8); - providing a separate flow of heat transfer medium through each compartment by means of a heat transfer medium inlet (12) and a heat transfer medium outlet (13) for each compartment, thereby regulating a separate temperature within each compartment; - directing the PCR solution through the tube (6), thereby directing the PCR solution through the distinct temperatures in each compartment, and preparing a DNA product based on the PCR solution; A method comprising:

[0211] 41. The method according to any of the previous embodiments, wherein the first temperature in the first compartment (7) is within the range of about 85°C to about 105°C, preferably about 98°C. 42. The method according to one of embodiments 40-41, wherein the second temperature in the second compartment (8) is in the range of about 45°C to about 72°C, preferably about 60°C to about 72°C.

[0212] 43. The method according to one of embodiments 40 to 42, further comprising providing a third section (15), wherein the tube (6) is wound from at least the second section (8) into the third section (15), and the third temperature in the third section (15) is within the range of about 65°C to about 75°C, preferably about 72°C.

[0213] 44. The method according to one of embodiments 40 to 43, wherein a first temperature in the first compartment (7) is set for a denaturation step of the polymerase chain reaction, and a second temperature in the second compartment (8) is set for an annealing step of the polymerase chain reaction, and optionally for an extension step of the polymerase chain reaction.

[0214] 45. The method according to embodiment 43, wherein a third temperature in the third compartment (15) is set for an extension step of the polymerase chain reaction. 46. ​​Use of a device according to one of embodiments 1 to 26 for preparing a DNA product by capillary polymerase chain reaction.

[0215] 47. Use of a manufacturing device (10) according to one of embodiments 27 to 38 for the production of a pharmaceutical product. A set of preferred embodiments of the present application relate to the following:

[0216] 1. A device (1) for amplifying DNA by polymerase chain reaction (PCR), - a tube (6) for conducting the PCR liquid; a first compartment (7), a second compartment (8), a third compartment (15), a tube (6) being wound through a first compartment (7) from the first compartment (7) to a second compartment (8), through the second compartment (8) from the second compartment (8) to a third compartment (15), and through the third compartment (15) from the third compartment (15) back to the first compartment (7), the tube (6) wound from compartment to compartment forming a helical stack of tube turns, each tube turn representing one PCR cycle, the tube (6) comprising a PCR fluid inlet (20) and a DNA product outlet (25); A device (1), wherein a first compartment (7), a second compartment (8) and a third compartment (15) each comprise a means for adjusting the temperature within the compartment to prepare a DNA product based on a PCR solution, the first compartment being configured to provide a temperature for denaturation, the second compartment (8) being configured to provide a temperature for annealing and the third compartment (15) being configured to provide a temperature for extension.

[0217] 2. A device (1) for amplifying DNA as described in embodiment 1, wherein the PCR liquid inlet (20) is connectable to a pump unit for pumping PCR liquid through the tube (6).

[0218] 3. The device (1) for amplifying DNA according to embodiment 1 or 2, wherein a partition (14) is disposed between the compartments and separates them from each other, the partition (14) having a through hole for the tube (6) to extend from one compartment to the next.

[0219] 4. A 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 a means for regulating the temperature within each compartment.

[0220] 5. The device (1) for amplifying DNA according to any one of embodiments 1 to 3, wherein each compartment comprises a heat medium inlet (12) and a heat medium outlet (13) as a means for adjusting the temperature within each compartment, i.e., by providing separate heat medium flows through the first compartment (7), the second compartment (8), and the third compartment (15) to adjust the separate temperatures within the first compartment (7), the second compartment (8), and the third compartment (15).

[0221] 6. The device (1) for amplifying DNA according to any one of the previous embodiments, wherein the tubes (6) extend in a wavy manner inside each of the compartments. 7. A device (1) for amplifying DNA according to any one of the previous embodiments, wherein each compartment forms a reservoir filled with a thermal medium or solid.

[0222] 8. A device (1) for amplifying DNA according to any one of the previous embodiments, further comprising at least a stand (17) arranged in one of the compartments to provide a fixing point for the tube (6) and / or a deflection point for the corrugation of the tube (6).

[0223] 9. The device (1) for amplifying DNA according to embodiment 8, wherein the stand (17) is replaceable and / or rearrangeable within the compartment. 10. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the tube (6) has a diameter in the range of about 0.5 mm to about 50 mm, preferably about 0.5 mm to about 1.5 mm, and more preferably about 0.75 to about 1.25 mm.

[0224] 11. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the tube (6) has a length between the tube inlet and the tube outlet in the range of about 10 m to about 200 m, preferably about 50 m to about 100 m.

[0225] 12. The device (1) for amplifying DNA according to any one of the preceding embodiments, wherein the tube (6) is sized to accommodate a PCR solution in the range of about 10 mL to about 500 mL, preferably about 20 mL to about 100 mL.

[0226] 13. The device for amplifying DNA (1) according to any one of the previous embodiments, wherein the spiral stack of tube turns comprises from about 10 to about 50 turns, preferably from about 15 to about 40 turns.

[0227] 14. A manufacturing device (10) comprising a device (1) for amplifying DNA according to any one of embodiments 1 to 13. 15. A manufacturing device (10) according to embodiment 14, further comprising a processing chamber (2) and a technology chamber (3).

[0228] 16. The manufacturing device (10) of embodiment 15, further comprising a control unit configured to control the gas flow through the processing chamber (2) as a gas shower. 17. A manufacturing device (10) according to embodiment 15 or 16, further comprising a separation element (4) separating the processing chamber (2) from the technical chamber (3), the separation element (4) comprising an attachment (16) protruding in the direction of the technical chamber (3) having an opening towards the processing chamber (2), and a device (1) for amplifying DNA can be inserted into the attachment (16).

[0229] 18. A manufacturing device (10) as described in embodiment 17, wherein when the device (1) for preparing a DNA product is inserted into the attachment (16), the front surface of the device (1) for preparing a DNA product terminates essentially flush with the opening of the attachment (16) into the processing chamber (2).

[0230] 19. A manufacturing module (100) comprising a manufacturing device (10) described in any one of embodiments 14 to 18, and at least one manufacturing device selected from the group consisting of a manufacturing device (10) having a processing medium supply unit and a mixing unit, a manufacturing device (10) having a purification unit, and a manufacturing device (10) having a filtration unit.

[0231] 20. A method for amplifying DNA by polymerase chain reaction (PCR), comprising: - providing a first section (7), a second section (8), a third section (15) and a tube (6), the tube (6) being wound from the first section (7) to the second section (8), from the second section (8) to the third section (15) and from the third section (15) to the first section (7) through the first section (7), the tube (6) wound from section to section forming a helical stack of tube windings; - providing means for regulating the separate temperature in each compartment; - directing the PCR solution through the tube (6), thereby directing the PCR solution through the distinct temperatures in each compartment, and preparing a DNA product based on the PCR solution; A method comprising:

[0232] 21. The method according to embodiment 20, wherein the temperature in the first compartment (7) is for denaturation and is preferably within the range of about 85°C to about 105°C, preferably about 98°C. 22. The method according to embodiment 20 or 21, wherein the temperature in the second compartment (8) is for annealing and is preferably within the range of about 45°C to about 72°C, preferably about 60°C to about 72°C.

[0233] 23. The method according to any one of embodiments 20 to 22, wherein the temperature in the third compartment (15) is for extension and is preferably within the range of about 65°C to about 75°C, preferably about 72°C.

[0234] 24. Use of the device (1) according to any one of embodiments 1 to 13 for preparing a DNA product by polymerase chain reaction. 25. Use of a manufacturing device (10) according to any one of embodiments 14 to 18 or a manufacturing module (100) according to embodiment 19 for the production of a pharmaceutical product. [Explanation of symbols]

[0235] 1. Device for preparing DNA products 2 Processing chamber 3. Technology Chamber 4 Separation elements 5. Housing 6 Tubes 7 First Section 8 Second Section 9 Duct 9a Gas inlet duct 9b Gas outlet duct 10 Manufacturing Devices 11 Collection Tray 12 Heat medium inlet 13 Heat medium outlet 14 Bulkhead 15 Third Section 16 Attachment 17 Stand 18 Combined Unit 19 Mounting elements 20 PCR liquid inlet 21 Sensors 22 Handle 23 PCR fluid intake 24 DNA product exit port 25 DNA product outlet 26 boxes 27 Replacement rail 28 Isolation section 29 Exterior wall 30 Internal partition wall 31 Heating unit 32 Heat pipe (part of the cooling unit) 33 Thermoelectric cooler (part of the cooling unit) 34 Ventilator 35 Air inlet 36 Electrical Plug 37 Pump unit for pumping PCR liquid 100 Manufacturing Modules

Claims

1. A device (1) for amplifying DNA by polymerase chain reaction (PCR), comprising: A tube (6) for conducting a PCR liquid; A first section (7), A second compartment (8); A third compartment (15), 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), the tube (6) wound from compartment to compartment forming a helical stack of tube turns, each tube turn representing one PCR cycle, the tube (6) comprising a PCR fluid inlet (20) and a DNA product outlet (25); The first compartment (7), the second compartment (8) and the third compartment (15) each comprise a means for adjusting the temperature in the compartment to prepare a DNA product based on a PCR solution, the first compartment being configured to provide a temperature for denaturation, the second compartment (8) being configured to provide a temperature for annealing, and the third compartment (15) being configured to provide a temperature for extension; A device (1) in which partitions (14) are disposed between the compartments to separate them from one another, said partitions (14) having through holes for said tubes (6) to extend from one compartment to the next.

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

3. Each compartment is provided with a heating unit (31) as a means for adjusting the temperature within each compartment. ) and optionally a cooling unit (32, 33), or 3. The device (1) for amplifying DNA according to claim 1 or 2, wherein each compartment comprises a heat medium inlet (12) and a heat medium outlet (13) as a means for regulating the temperature within each compartment, i.e. for providing separate heat medium flows through the first compartment (7), the second compartment (8) and the third compartment (15) to regulate separate temperatures within the first compartment (7), the second compartment (8) and the third compartment (15).

4. The device (1) for amplifying DNA according to any one of claims 1 to 3, wherein the tube (6) extends in a wavy manner inside each of the compartments.

5. A device (1) for amplifying DNA according to any one of claims 1 to 4, wherein each compartment forms a reservoir which is filled with a thermal medium or a thermal solid.

6. The device (1) for amplifying DNA according to any one of claims 1 to 5, further comprising at least a stand (17) arranged in one of the compartments to provide a fixing point for the tube (6) and / or a deflection point for the corrugation of the tube (6).

7. 7. The device (1) for amplifying DNA according to claim 6, wherein the stand (17) is exchangeable and / or repositionable within the compartment.

8. the tube (6) has a diameter in the range of about 0.5 mm to about 50 mm; and / or the tube (6) has a length between the tube inlet and the tube outlet in the range of about 10 m to about 200 m; and / or The device (1) for amplifying DNA according to any one of claims 1 to 7, wherein the tube (6) is dimensioned to accommodate a PCR solution in the range of about 10 mL to about 500 mL.

9. The device (1) for amplifying DNA according to any one of claims 1 to 8, wherein the helical stack of tube turns comprises from about 10 to about 50 turns.

10. A manufacturing device (10) comprising a device (1) for amplifying DNA according to any one of claims 1 to 9, further comprising: A processing chamber (2) and a technology chamber (3), a control unit configured to control the gas flow through the treatment chamber (2) as a gas shower; A manufacturing device (10).

11. 11. The manufacturing device (10) of claim 10, further comprising a separation element (4) separating the processing chamber (2) from the technical chamber (3), the separation element (4) comprising an attachment (16) protruding in the direction of the technical chamber (3) with an opening towards the processing chamber (2), and a device (1) for amplifying DNA can be inserted into the attachment (16).

12. 1. A method for amplifying DNA by polymerase chain reaction (PCR), comprising: providing a first section (7), a second section (8), a third section (15), and a tube (6), the tube (6) being wound from the first section (7) to the second section (8), from the second section (8) to the third section (15), and from the third section (15) to the first section (7), the tube (6) wound from section to section forming a spiral stack of tube windings, partitions (14) being disposed between the sections separating them from one another, the partitions (14) comprising through holes for the tube (6) to extend from one section to the next; providing means for regulating separate temperatures within each compartment; directing a PCR solution through said tube (6), thereby directing the PCR solution through distinct temperatures in each compartment, and preparing a DNA product based on the PCR solution; A method comprising:

13. the temperature in the first compartment (7) is for denaturation and is in the range of about 85°C to about 105°C; and / or the temperature in the second compartment (8) is for annealing and is in the range of about 45°C to about 72°C; and / or The method of claim 12, wherein the temperature in the third section (15) for extension is in the range of about 65°C to about 75°C.

14. Use of the device (1) according to any one of claims 1 to 9 for preparing a DNA product by polymerase chain reaction.

15. 12. Use of a manufacturing device (10) according to claim 10 or 11 for the production of a pharmaceutical product.

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