Methods and Assemblies for Operating a Continuous Downstream Process - Patent application

Abstract

The present invention relates to a method for operating a continuous downstream process, in particular a continuous viral inactivation process or an in-line dilution process, using a bioprocess assembly (1), comprising a compensating vessel (2) having a first fluid inlet (3) for introducing a concentrate stream, in particular a product or buffer concentrate stream, into said compensating vessel (2), at least one second fluid inlet (4) for introducing a diluent stream, in particular a product or buffer diluent stream, into said compensating vessel (2), and a fluid outlet (5) for leading a liquid stream out of said compensating vessel (2), the bioprocess assembly (1) comprising a transport assembly (6) for fluid transport, the transport assembly (6) being assigned to the first and second fluid inlets (3) and (4), respectively, for transporting at least one liquid stream to the compensating vessel (2) and the fluid outlet (5) for transporting a liquid stream from said compensating vessel (2), and the bioprocess assembly (1) comprising an electronic process control device (7). It is proposed that the electronic process control device (7) adapts the filling level in the compensation container (2) by open-loop and / or closed-loop control of said transport assembly (6) so that it does not fall below a predefined minimum filling level in the compensation container (2).

Description

[Technical Field]

[0001] The present invention relates to a method for operating a continuous downstream process according to the preamble of claim 1, a bioprocess assembly for operating a continuous downstream process according to claim 25, an electronic process controller for the proposed bioprocess assembly according to claim 26, a use of the electronic process controller for carrying out the proposed method according to claim 28, a computer program for the proposed electronic process controller according to claim 29, and a computer-readable storage medium for storing a computer program according to claim 30. [Background technology]

[0002] The method for operating a continuous downstream process, in particular a continuous viral inactivation process or an in-line dilution process, is used in the context of bioprocesses for the production and / or quality control of biopharmaceutical bioproducts, in particular proteins, vectors, cells and the like, whereby a continuous flow from an upstream unit, in particular an upstream chromatography unit, can be provided to a downstream unit, in particular a downstream chromatography unit or filtration unit.

[0003] In process engineering, the term "downstream process" refers to all methods used to separate and purify fermentation products from the fermentation broth of a biotechnology process. This term includes mechanical, thermal, electrical, and physicochemical methods.

[0004] The term "bioprocess" herein refers to biotechnological and biopharmaceutical processes involved in the production of therapeutic bioproducts, such as vaccines, biologics, components for cell or gene therapy, or non-therapeutic bioproducts, such as pigments, biofuels, or nutrients. Such bioproducts can either be produced by living cells, or the cells themselves can be the bioproduct, or the bioproduct can be the result of cell-free production based on cellular components that are either naturally or non-naturally occurring.

[0005] The trend toward process intensification in the biopharmaceutical industry is leading to the increasingly frequent implementation of continuous process steps. In batch production, as performed in a routine and standardized manner, the product passes through successive process steps in a batch-by-batch fashion and is collected in a storage vessel after each step. This approach can limit both the utilization of the bioprocess assembly and the productivity of the process. On the other hand, continuous process control means that the product passes through multiple manufacturing steps without interruption. Thus, continuous methods allow individual work steps to be performed on a smaller scale, thus saving consumables such as buffers, solvents, and the like, and allowing even large processes to be performed in a single-use format. In addition, the reduction in the number and size of storage vessels also shortens the residence time of the product in the bioprocess.

[0006] The advantages of continuous manufacturing lie in, among other things, previously unachieved control of product quality and greater agility and flexibility in addressing product requirements. A further advantage is that the risks of scale-up are significantly reduced due to the compact plant design. Continuous manufacturing therefore has great potential to increase the economic efficiency of bioprocessing as a whole.

[0007] As part of process intensification, new methods such as single-use centrifugation, tangential flow filtration (TFF) or rapid cycling chromatography (RCC) are increasingly being developed, all with the aim of achieving a continuous product flow and thus minimizing the space and costs of the process steps used in downstream processes.

[0008] The first chromatographic step, particularly the Protein A affinity chromatography step, is routinely followed by the first viral inactivation step. The implementation of successive viral inactivation steps presents a process development challenge as part of process intensification.

[0009] The known method for operating a continuous downstream process (EP 2867359), on which the present invention is based, is used for semi-continuous virus inactivation during the protein production process, eliminating the need for large storage vessels for virus inactivation. This is based on the fact that smaller stirred mixing vessels are used for virus inactivation, thereby creating better conditions for the product and saving installation space. Furthermore, the incubation time for complete virus inactivation can be reduced from at least one hour to a maximum of one hour.

[0010] However, this method is not a continuous process because the Protein A eluate is incubated with the viral inactivation agent using a mixing vessel, which can still require up to an hour for complete viral inactivation. During this time, the Protein A eluate containing the target protein must be incubated and stirred with the viral inactivation agent for an extended period of time, similar to a traditional batch process, before proceeding to the next step. This can be problematic because excessive storage or processing times can destroy sensitive bioproducts, such as antibodies, from bioprocesses. The longer a bioproduct, especially a target protein, is incubated with the viral inactivation agent, the greater the chance that it will be destroyed by the mechanical force of the stirrer and the degradative effects of the viral inactivation agent.

[0011] Similar challenges arise when performing a continuous in-line dilution process in which two or more liquids are mixed together. Currently, mixing of, for example, buffers or media is often performed batchwise, especially in downstream processes. The base liquid to be mixed is filled into a container, followed by the addition of the components to be incorporated and mixing in a mixer until the desired degree of mixing or solubility is achieved, whereby the completed solution is then further processed.

[0012] However, again in this case, similar to the problem of viral inactivation explained above, the use of such intermediate vessels has the crucial drawback that the process sequence is interrupted by the mixing process carried out batchwise using the mixing vessel, which involves the consumption of more time, space and assembly, which in turn leads to increased costs and reduces the economic efficiency of the bioprocess.

[0013] For this reason, the mixing process should preferably be realized "in-line," i.e., within the pipeline of the pipeline assembly, rather than in an intermediate container; therefore, currently, static mixing elements are preferably used for in-line mixing. Known methods for operating continuous downstream processes, particularly continuous in-line dilution processes (WO 2015 / 117884), include a first and at least one second chromatography column, multiple buffer containers, and an additional in-line dilution system with an inlet and a bioprocess outlet. However, the additional in-line dilution system requires additional manufacturing space, resulting in increased material costs, increased structural complexity, and increased maintenance costs.

[0014] As a result, known methods offer limited efficiency, on the one hand, in terms of long process times that endanger the biological product, and on the other hand, in terms of production costs and production space utilization. Generally, the implementation of known methods for operating continuous downstream processes, in particular continuous viral inactivation processes or in-line dilution processes, is relatively laborious. Summary of the Invention [Problem to be solved by the invention]

[0015] The problem underlying the present invention is to configure and further develop known methods for operating continuous downstream processes in such a way that their implementation is simplified. [Means for solving the problem]

[0016] The above problem is solved in a method for operating a continuous downstream process according to the preamble of claim 1 by the features of the characterizing part of claim 1.

[0017] The key idea is that the electronic process control device adapts the fill level in the compensation vessel by open-loop and / or closed-loop control of the transport assembly to avoid falling below a minimum fill level, thereby ensuring a continuous liquid flow from the bioprocess assembly.

[0018] The term "compensation container" should be understood broadly in this context and here refers to any container or receptacle intended to hold a liquid. It may be made of flexible or rigid material, in particular metal, glass, plastic or the like. Additionally or alternatively, the compensation container may be configured for multiple uses or as a single-use container.

[0019] The special design of the proposed method for operating a continuous downstream process has the advantage of converting a particularly discontinuous liquid stream from an upstream process step, in particular an upstream Protein A chromatography step, which may fluctuate in terms of volume and product concentration, i.e., there may be times when the liquid stream is not flowing at all and the product concentration may usually fluctuate during the chromatography stage, into a continuous liquid stream and continuously conducting this liquid stream into the proposed continuous downstream process, in particular a continuous virus inactivation process or an in-line dilution process followed by a downstream process step.

[0020] The term "continuous liquid flow" herein means a liquid flow that is not interrupted, for example by incubation in a vessel or the like, but that may vary in flow rate.

[0021] In particular, it is proposed that the electronic process control device adapts the filling level in the compensation container by open-loop and / or closed-loop control of the transport assembly so that it does not fall below a predefined minimum filling level in the compensation container.

[0022] According to a preferred feature of claim 2, the electronic process control device adapts the filling level in the compensation container so as to achieve a predetermined target filling level or to maintain the filling level within a target filling level range. This allows the user not only to prevent the compensation container from running empty, but also to additionally set a predetermined target filling level, which is preferably above the minimum filling level, in particular after the filling level has fallen below or exceeded the target filling level.

[0023] A preferred configuration according to claim 3 or 4 relates to the details of at least one conveying device assigned to the conveying assembly, which allows flexible fluid controllability in a simple manner.

[0024] The term "conveying assembly" here should be understood broadly in the present context and refers here to any kind of assembly provided for conveying and / or adapting, in particular interrupting, a liquid, in particular a pump assembly with at least one pump and / or a valve assembly with at least one valve. In this case, such a pump and such a valve each form a "conveying device" of the conveying assembly. Advantageously, such a conveying device is intended to be a metering pump, in particular one configured as a tube pump, rotary piston pump or diaphragm pump. In particular, such a valve is intended to be provided for establishing a selective and quantitative fluid connection, advantageously a continuous valve, a regulating valve or the like.

[0025] According to a preferred feature of claim 5, the electronic process control device derives an actual value of a fill level parameter in the compensation container from the process data during the monitoring routine. Depending on the deviation between the derived actual value of the fill level parameter and a predetermined minimum value of the fill level parameter corresponding to a predetermined minimum fill level in the compensation container, the electronic process control device adapts the conveying capacity of the first, second, and / or third conveying devices to prevent the minimum fill level from being exceeded. This feature allows the electronic process control device to react to any deviation from the minimum fill level and adapt the corresponding pump to prevent the minimum fill level from being exceeded. Alternatively, depending on the deviation between the derived actual value of the filling level parameter and a predetermined target value of the filling level parameter corresponding to a predetermined target filling level in the compensation container, an adaptation of the conveying capacity of the first, second and / or third conveying device is performed to achieve the target filling level, or depending on the deviation between the derived actual value of the filling level parameter and a predetermined limit value of a predetermined target filling level range in the compensation container, an adaptation of the conveying capacity of the first, second and / or third conveying device is performed to maintain the filling level within the target filling level range.

[0026] According to a particularly preferred feature of claim 6, the electronic process control device generates a first model from the process data. The degree of adaptation of the transport capacities is derived from this first model. This feature provides the advantage that not only is the method controllable by adapting the transport capacities of the first, second and / or third transport devices, but this adjustment can also be performed automatically by means of the underlying first model.

[0027] The first model represents, based on the included process data, the relationship between the fill level parameter in the compensation vessel on the one hand and the conveying capacity of the first, second and / or third conveying device on the other hand.

[0028] According to a preferred configuration of claim 7, the bioprocess assembly has a first bioprocess inlet and a fourth conveying device assigned thereto, which is provided for metering a liquid flow into the pipeline assembly. Further liquid flows can be supplied via additional bioprocess inlets. This configuration opens up the possibility of generating reactive liquid flows for further parameter adaptation in a particularly simple manner, without compromising performance in other respects.

[0029] According to a further preferred configuration according to claim 8, the concentrate stream is combined with the diluent stream in the compensation vessel to form a first liquid stream containing the target agent, which is subsequently combined with the second liquid stream in a precisely predefined volume ratio to form a third reactive liquid stream. This simple process control makes it particularly easy to create predefined conditions, in particular virus inactivation conditions or buffer dilution conditions.

[0030] According to a further preferred configuration of claim 9, the bioprocess assembly comprises at least one mixer for mixing the third reactive liquid stream, which allows for a particularly simple homogenization of the target parameters, in particular the pH or conductivity.

[0031] A particularly preferred configuration according to claim 10 shows that the bioprocess assembly has a residence time assembly downstream of the mixer in order to provide a minimum residence time for the liquid flow, which in a particularly simple way ensures the required incubation time of the liquid flow, in particular with the virus inactivation agent.

[0032] According to further preferred configurations of claims 11 and 12, the sensor assembly generates process data into which liquid parameters of the first liquid stream containing the target factor and liquid parameters of the third reactive liquid stream are input. Based on the actual values ​​of the liquid parameters determined by the measurement, the electronic process control device performs a virus inactivation routine in order to adapt the target values ​​of the liquid parameters, in particular the target pH or target conductivity, in the liquid stream. This makes it possible to constantly adapt otherwise deviating values, in particular by adapting the acid concentration or the buffer concentration, in order to keep them as constant as possible, thereby enabling particularly efficient continuous virus inactivation or continuous in-line dilution.

[0033] A further preferred configuration according to claim 13 relates to a waste line provided for conducting a liquid flow from the bioprocess assembly. This variant offers the advantage that a liquid flow or part thereof that should not be conducted further through the bioprocess assembly can be diverted, in particular withdrawn from the bioprocess assembly for further use or discarded. This allows so-called priming of the system, i.e., injection of buffer into the line assembly, discarding (partial) liquid flows and / or the like that are no longer required, and generally increases the flexibility of the proposed method. For these purposes, according to claim 14, a valve assembly is provided, which in its other aspects comprises at least one valve, for selective fluid connection of each line with the line assembly.

[0034] A further preferred configuration according to claim 15 relates to a configuration of the bioprocess assembly with a second bioprocess inlet to which a fifth conveying device is assigned, whereby a fourth neutralizing liquid stream is introduced and combined with the third reactive liquid stream to generate a fifth combined liquid stream, which offers the advantage that neutralization of virus inactivation conditions in particular is particularly simple.

[0035] A particularly preferred configuration of the conveying device for the metered delivery of the liquid stream is given in claim 16 .

[0036] In a further preferred configuration according to claim 17, the bioprocess assembly comprises at least one second mixer provided for mixing the fifth composite liquid stream, which allows for particularly simple homogenization of the neutral conditions, in particular the pH, and allows for continuous operation of the process.

[0037] Preferred configurations according to claims 18 and 19 relate to preferred measurement positions and measurement parameters of the sensors of the sensor assembly, so that process data can be generated particularly simply and efficiently. Furthermore, the electronic process control device implements a neutralization routine, thereby adapting a target value for a liquid parameter in the liquid flow, in particular pH or conductivity. This allows otherwise deviating values ​​to be constantly adapted, in particular by adapting the base concentration or the buffer concentration, in order to keep them as constant as possible, thereby enabling particularly efficient continuous virus inactivation or continuous in-line dilution.

[0038] According to a further preferred configuration as set forth in claim 20, the electronic process control device generates a second model from the process data, and the degree of adaptation of the conveying capacity of the fourth and / or fifth conveying device is derived from this second model. This configuration offers the advantage that not only is the method adjustable, but this adjustment can also be performed automatically by means of the underlying second model, for particularly simple implementation of continuous virus inactivation processes or in-line dilution processes.

[0039] According to a preferred feature of claim 21, the first model and / or the second model are statistical or analytical models, which ensures particular flexibility and efficiency in the applicability of the proposed method.

[0040] The preferred configuration according to claim 22 relates to a continuous outflow of the liquid flow from the bioprocess assembly, which provides a particularly simple justification for the connectability of the proposed process with downstream, preferably continuous, processes.

[0041] A preferred configuration according to claim 23 relates to the configuration of the first and / or second mixer. Preferably, a non-positive displacement pump is used for dynamic in-line mixing, which is driven in a reverse flow direction compared to the normal operation. This configuration opens up the possibility that the impeller of the non-positive displacement pump, which rotates in the opposite direction to the normal direction, can also function as a mixer, which makes it possible to set a variable mixing capacity by adjusting the rotation speed without causing a pressure increase in the system.

[0042] According to an equally preferred feature of claim 24, the proposed method is carried out in combination with a chromatographic and / or filtration method, which offers the possibility of simply integrating the proposed method into existing processes.

[0043] According to a further teaching of independent claim 25, patent protection is claimed for a bioprocess assembly for operating a continuous downstream process, in particular a continuous virus inactivation process or an in-line dilution process, during a bioprocess, the bioprocess assembly comprising a compensating vessel having a first fluid inlet for introducing a concentrate stream, in particular a product or buffer concentrate stream, into the compensating vessel, at least one second fluid inlet for introducing a diluent stream, in particular a product or buffer diluent stream, into the compensating vessel, and a fluid outlet for leading a liquid stream from the compensating vessel; the bioprocess assembly comprises a transport assembly for fluid transport, the transport assembly being assigned to the first and second fluid inlets, respectively, for transporting at least one liquid stream to the compensating vessel and to the fluid outlet for transporting the liquid stream from the compensating vessel; and the bioprocess assembly comprises an electronic process control device. In this regard, reference can be made to the full description of the proposed method for operating a continuous downstream process.

[0044] In particular, it is proposed that during normal operation of the bioprocess assembly, the electronic process control device adapts the fill level in the compensation container by open-loop and / or closed-loop control of the transport assembly so that it does not fall below a predefined minimum fill level in the compensation container.

[0045] According to the further teaching of claim 26, which has an independent meaning, patent protection is claimed for an electronic process control device of the proposed bioprocess assembly. In this regard, reference may be made to the entire description of the proposed method for operating a continuous downstream process and the proposed bioprocess assembly.

[0046] According to a preferred configuration according to claim 27, the electronic process control device comprises a data processing system for implementing the proposed method.

[0047] According to the further teaching of claim 28, which has an independent meaning, patent protection is claimed for the use of an electronic process control device for carrying out the proposed method. In this regard, reference may be made to all descriptions relating to the proposed method for operating a continuous downstream process, the proposed bioprocess assembly, and the proposed electronic process control device.

[0048] According to the further teaching of claim 29, which has an independent meaning, patent protection is claimed for a computer program for the proposed electronic process control device. In this regard, reference may be made to all descriptions relating to the proposed method for operating a continuous downstream process, the proposed bioprocess assembly, the proposed electronic process control device and the proposed use.

[0049] According to a further teaching according to claim 30, which has an independent meaning, patent protection is claimed for a computer-readable storage medium for storing the proposed computer program. In this regard, reference may be made to all descriptions relating to the proposed method for operating a continuous downstream process, the proposed bioprocess assembly, the proposed electronic process control device, the proposed use, and the proposed computer program.

[0050] In the following, the invention will be explained in more detail with reference to drawings which show only example configurations. [Brief explanation of the drawings]

[0051] [Figure 1] FIG. 1 shows an example configuration of the proposed method for operating a continuous downstream process. [Figure 2] FIG. 1 shows a further example configuration of the proposed method for operating a continuous downstream process, in particular a continuous viral inactivation process. [Figure 3] FIG. 2 shows a flow chart illustrating the preferred operating principle of the method proposed according to FIG. 1. DETAILED DESCRIPTION OF THE INVENTION

[0052] Figure 1 illustrates a proposed method for operating a continuous downstream process, particularly a continuous viral inactivation process or an in-line dilution process. This is used in downstream processes of bioprocesses using the bioprocess assembly 1, particularly in the production, purification, and / or quality control of biopharmaceutical products, e.g., during protein production. Such proteins may be growth factors, hormones, enzymes, particularly antibodies, antibody derivatives, or the like. The proposed method can be used to ensure that biopharmaceutical products are free of any type of active viral particles above a certain threshold, e.g., defined by the manufacturer and / or regulatory authorities, and in particular completely free of active viral particles. Alternatively, the proposed method can be used for continuous in-line dilution, particularly of buffer solutions.

[0053] The target protein may originate directly or indirectly from the bioreactor, in particular after a downstream process step has been carried out, such as a processing step, in particular a filtration, precipitation, and / or chromatographic separation step or the like. Such a chromatographic step may for example be an affinity chromatography step, in particular an affinity chromatography step using Protein A.

[0054] The bioprocess assembly 1 comprises a compensation vessel 2 with a first fluid inlet 3 provided for introducing a concentrate stream, in particular a product or buffer concentrate stream, into the compensation vessel 2, and at least one second fluid inlet 4 provided for introducing a diluent stream, in particular a product or buffer diluent stream, into the compensation vessel 2. The compensation vessel 2 here is advantageously a stirred compensation vessel 2 as shown in Figures 1 and 2 and is therefore provided for homogenization.

[0055] The term "product concentrate stream" as used herein means a liquid stream that essentially comprises a product from an upstream process step, particularly an eluate from an upstream chromatography step.

[0056] By "buffer concentrate stream" herein is meant a liquid stream that essentially comprises a buffer, particularly a concentrated buffer, that is diluted in-line for further use in a bioprocess.

[0057] The term "product diluent stream" herein means a liquid stream essentially comprising a solvent intended to dilute the product from an upstream process step, in particular a buffer appropriate for the respective product.

[0058] The term "buffer diluent stream" herein means a liquid stream essentially comprising a solvent, particularly water, provided to dilute the buffer.

[0059] The bioprocess assembly 1 has a fluid outlet 5 provided for leading out a liquid flow from the compensation vessel 2. Furthermore, the bioprocess assembly 1 has a transport assembly 6 for fluid transport, the transport assembly 6 being assigned to the first and second fluid inlets 3 and 4 for transporting at least one liquid flow to the compensation vessel 2, respectively, and to the fluid outlet 5 for transporting a liquid flow from the compensation vessel 2. Furthermore, the bioprocess assembly 1 has an electronic process control device 7.

[0060] The term "fluid transport" should be understood broadly in the present context and means any kind of transport and / or adaptation, especially interruption of transport, of a fluid, especially a liquid.

[0061] Here, it is essential in the proposed method that the electronic process control device 7 adapts the filling level in the compensation vessel 2 by open-loop and / or closed-loop control of the transport assembly 6 so that it does not fall below a predefined minimum filling level in the compensation vessel 2. This ensures a continuous liquid flow that can be delivered from the bioprocess assembly 1 (FIG. 1).

[0062] The term "minimum filling level" is to be understood broadly in the present context and advantageously means any kind of filling level in the compensation container, advantageously defined by the user as the minimum filling level. According to a particularly preferred configuration, the minimum filling level is advantageously less than 50%, more advantageously less than 25%, even more advantageously less than 10%, even more advantageously less than 5% of the total volume of the compensation container 2.

[0063] Advantageously, the electronic process control device 7 adapts the filling level in the compensation container 2 by open-loop and / or closed-loop control of the transport assembly 6 to achieve a predefined target filling level or to maintain the filling level within a target filling level range.

[0064] The term "target filling level" here means any filling level, preferably defined by the user, as the filling level to be achieved. The term "target filling level range" means a range between upper and lower filling level limits within which the filling level is to be maintained.

[0065] Advantageously, the conveying assembly 6 comprises a first conveying device 8 for fluid conveyance, which is assigned to the first fluid inlet 3. Additionally or alternatively, the conveying assembly 6 comprises a second conveying device 9 for fluid conveyance, which is assigned to the second fluid inlet 4. Additionally or alternatively, the conveying assembly 6 comprises a third conveying device 10 for fluid conveyance, which is assigned to the fluid outlet 5.

[0066] Advantageously, the bioprocess assembly 1 comprises a bioprocess outlet 11 provided for withdrawing liquid from the bioprocess assembly 1. Furthermore, the bioprocess assembly 1 comprises a conduit assembly 12 with at least one conduit 13 for fluidly connecting the fluid outlet 5 of the compensation vessel 2 to the bioprocess outlet 11. This withdrawal from the bioprocess outlet 11 advantageously occurs when a liquid stream passes through the proposed method and is led to a downstream process step, advantageously a downstream chromatography step or the like, for further processing.

[0067] A third conveying device 10 is now advantageously provided for fluid conveyance through at least one line 13 of a line assembly 12, as can be seen in FIGS.

[0068] Furthermore, the bioprocess assembly 1 comprises a sensor assembly 14 for generating process data 15 , comprising at least one sensor 16 assigned to the compensation vessel 2 .

[0069] At least one sensor 16 in the compensation container 2 is preferably configured as a filling level sensor, which is further preferably provided for measuring the filling level and / or for measuring the change in filling level per unit time, and further preferably for measuring a filling level parameter.

[0070] The term "filling level parameter" is understood here in a broad sense. The filling level parameter is advantageously generally a measure for filling the compensation container 2 with liquid and can be expressed by the filling rate of the compensation container 2, a specific mass, a specific volume, a specific concentration, etc. Furthermore, the filling level parameter can be expressed as a specific filling level at a specific time or as a change in the filling level over time, and thereby also as a flow rate.

[0071] The term "liquid" is likewise understood in a broad sense: it includes not only pure liquids but also emulsions and suspensions, for example heterogeneous mixtures of at least two different liquids or heterogeneous mixtures consisting of solid and liquid parts.

[0072] Additionally or alternatively, the at least one sensor 16 in the compensation container 2 may be configured as a force sensor, preferably a load cell, which is further preferably provided for measuring a force, in particular a mass, and / or a change in mass per unit time. Advantageously, this force, in particular a mass and / or a change in mass per unit time can be converted into a filling level and / or a change in filling level per unit time.

[0073] Advantageously, the electronic process control device 7 derives at least one actual value of a filling level parameter in the compensation container 2 from the process data 15 in a monitoring routine 17, advantageously in an information retrieval 18. Depending on a deviation, advantageously defined in a decision point 19, between the derived actual value of the filling level parameter and a predefined minimum value of the filling level parameter corresponding to a predefined minimum filling level in the compensation container 2, the electronic process control device 7 advantageously adapts, in an adaptation step 20, the conveying capacities of the first, second and / or third conveying devices 8, 9, 10 in order to prevent the minimum filling level from being exceeded, as shown in FIG.

[0074] Alternatively, depending on the deviation between the actual value of the derived filling level parameter and a predefined target value of the filling level parameter corresponding to a predefined target filling level in the compensation container 2, the electronic process control device 7 adapts the conveying capacities of the first, second and / or third conveying devices 8, 9, 10 to achieve the target filling level.

[0075] Alternatively, depending on the deviation between the actual value of the derived filling level parameter and a predefined limit value of a predefined target filling level range in the compensation container 2, the electronic process control device 7 adapts the conveying capacity of the first, second and / or third conveying devices 8, 9, 10 to maintain the filling level within the target filling level range.

[0076] The term "deviation" should be understood broadly in this context and includes not only any subtraction between a measured value and a target value, but also any other type of mathematical deviation, including variance, difference and / or integral of the difference or the like.

[0077] Advantageously, in the adaptation step 20, the conveying capacity of at least the first and / or second conveying device 8, 9 is adapted, advantageously increased. Additionally or alternatively, the conveying capacity of at least the third conveying device 10 is advantageously adapted, even more advantageously reduced.

[0078] 1 , the electronic process control device 7 derives a fill level parameter in the compensation container 2 from the process data 15 in a monitoring routine 17. When the electronic process control device 7 detects a deviation from a predetermined target fill level in the compensation container 2, the electronic process control device 7 advantageously adapts, and more advantageously increases, the conveying capacity of at least the first and / or second conveying devices 8, 9. This results in an increase in the liquid delivered to the compensation container 2, so that a continuous liquid flow out of the compensation container 2 can be maintained. Advantageously, the liquid flow supplied to the compensation container 2 to achieve the predetermined target fill level is a concentrate flow and / or a diluate flow. For this reason, the supply of a liquid flow to maintain a continuous liquid flow out of the compensation container 2 advantageously does not exceed a measure that would result in the concentration in the compensation container 2 falling below the target factor concentration, unless the continuous liquid flow out of the compensation container 2 would otherwise be stopped thereby.

[0079] The term "target factor" here means the product, preferably the eluate or the buffer. Similarly, the term "target factor concentration" here means the product concentration, preferably the eluate concentration or the buffer concentration. As already indicated above, it should be noted that the supply of the liquid stream to the compensation vessel 2 preferably does not exceed a measure below a predefined product concentration, preferably the eluate concentration or preferably the predefined buffer concentration. This can advantageously be set by the user of the proposed method before and / or during the method for operating a continuous downstream process.

[0080] Additionally or alternatively, the conveying capacity is advantageously adapted, and even advantageously reduced, of at least the third conveying device 10. As a result, the flow rate of the liquid flow leaving the compensation container 2 is indeed reduced, but it is possible to maintain a continuous liquid flow leaving the compensation container 2. It is particularly preferred to first adapt, and even advantageously increase, the conveying capacity of at least the first and / or second conveying device 8, 9 in order to maintain a continuous flow.

[0081] Advantageously, the conveying capacity of at least the third conveying device 10 is adapted if the adaptation of the conveying capacities of at least the first and / or second conveying devices 8, 9 is not sufficient to maintain a continuous liquid flow. Advantageously, the conveying capacity of the third conveying device 10 is kept substantially constant. Otherwise, changes in flow rate may result in the residence time of the liquid flow in the bioprocess assembly 1, which will be described in more detail below, falling below a predefined minimum residence time.

[0082] 1, 2 and 3, the electronic process control device 7 generates a first model 21 from the process data 15. A model here always means a mathematical model. The degree of adaptation of the conveying capacity of the conveying assembly 6 is preferably derived from the first model 21. The first model 21 further preferably represents the relationship between, on the one hand, a fill level parameter in the compensation container 2, preferably a change in the target fill level per hour, and, on the other hand, the conveying capacity of the first, second and / or third conveying device 8, 9, 10. Advantageously, the first model 21 is provided to predict a future target fill level in the compensation container 2 as a function of the change per hour, preferably at a predefined time point.

[0083] 2, the bioprocess assembly 1 advantageously has, downstream of the compensation vessel 2, a first bioprocess inlet 22 provided for introducing a liquid flow into the pipeline assembly 12 of the bioprocess assembly 1, which is advantageously arranged downstream of the third conveying device 10. The conveying assembly 6 advantageously has a fourth conveying device 23 assigned to the first bioprocess inlet 22 and provided for metering the liquid flow into the pipeline assembly 12.

[0084] The concentrated liquid stream is now advantageously combined with the diluent stream in the compensation vessel 2 to form a first liquid stream 24 containing target factors. Advantageously, this first liquid stream 24 containing target factors comprises either a biopharmaceutical target product, such as an antibody or a vector, or a process-relevant virus as target factor. Alternatively, this first liquid stream 24 advantageously comprises a target buffer, such as an equilibration buffer, a wash buffer or an elution buffer or the like, as target factor. In this respect, the term "target factor" therefore refers to a specific property of the liquid of the liquid stream 24, as already mentioned above.

[0085] A first liquid stream 24 comprising the target factor is advantageously introduced from the compensation vessel 2 through the fluid outlet 5 into at least one fluidly connected conduit 13 of the conduit assembly 12. Subsequently, it is advantageously combined in a predefined volume ratio with a second liquid stream 25 introduced through the first bioprocess inlet 22 and conveyed by a fourth conveying device 23 to form a third reactive liquid stream 26, thereby setting predefined target values ​​of fluid parameters, in particular pH and / or conductivity, in the third reactive liquid stream 26.

[0086] The second liquid stream 25 has the necessary characteristics for its intended function to satisfy viral inactivation conditions, particularly a pH of less than 3. The viral inactivation conditions, particularly the pH, of the second liquid stream 25 are selected so that after it is combined with the first liquid stream 24 containing the target agent, the resulting third reactive liquid stream 26 also has viral inactivation conditions, particularly a pH of 3 to 3.8 and / or a detergent concentration of 0.05% to 10% (v / v). Such conditions result in effective viral inactivation without damaging the respective products of the bioprocess, particularly proteins. The pH is achieved by adding an acid, such as lactic acid, ascorbic acid, acetic acid, hydrochloric acid, phosphoric acid, citric acid, glycine, succinic acid, and / or sulfuric acid, or the like. Advantageously, the viral inactivation reagent comprises an acid having a titratable group with a pKs of 2.0 to 4.3. The virus inactivation conditions can be selected so that the acid concentration can be up to 100 mM, yet still allow effective virus inactivation, while still having sufficient buffering properties to prevent damage to the protein product, for example, by acid denaturation. Additionally or alternatively, the virus inactivation conditions can be created by non-ionic detergents with chromophores. These can be, for example, Triton-X 100 and other polyethylene oxides.

[0087] The virus inactivation conditions are particularly important in the proposed method, regardless of whether it is exactly one specific virus type, multiple virus types, and / or multiple different virus types, and should not exceed 1 × 10 1times, advantageously at least 1 × 10 3 times, and even more advantageously at least 1×10 6 The viral inactivation conditions can be selected such that less than 1 ppm, advantageously less than 1 ppb of the volume of the third reactive liquid stream 26 is achieved at least 1×10 1 times, especially at least 1×10 6 The dwell time can be selected to be shorter than that required for effective viral inactivation.

[0088] 2, the second liquid stream 25 is advantageously mixed with the first liquid stream 24 containing the target protein for viral inactivation in a precise predefined volume ratio of these two liquid streams 24, 25 relative to each other, thereby ensuring that the conditions in the third reactive liquid stream 26 are actually provided and evenly distributed to perform the defined function. Such a volume ratio can be, for example, 19:1, with 19 parts of the first liquid stream 24 containing the target protein and 1 part of the second viral inactivation liquid stream 25.

[0089] Alternatively, the second liquid stream 25 has a buffer dilution condition as a necessary characteristic for performing its prescribed function, and advantageously the second liquid stream 25 consists of a dilution buffer and / or water or the like. In this alternative, the buffer dilution condition of the second liquid stream 25 is selected such that after being combined with the first liquid stream 24 containing the target factor, the resulting third reactive liquid stream 26 also has the buffer dilution condition, in particular the target buffer concentration. Dilution to create the target buffer concentration is achieved by adding a dilution buffer and / or water or the like.

[0090] Here, the bioprocess assembly 1 advantageously comprises a first mixer 27 provided for fluid mixing, advantageously downstream of the first bioprocess inlet 22. The third reactive fluid stream 26 is advantageously led through the first mixer 27 for fluid mixing. This mixer 27 is advantageously configured as an in-line static mixer, in particular as a radial static mixer or a laminar static mixer or the like, as shown in FIG. 2. Advantageously, several static mixers may be connected in series depending on process or precision requirements. Additionally or alternatively, a dynamic mixer may also be provided as the first mixer 27.

[0091] The term "fluid mixing" should be understood broadly in the present context and refers to any kind of mixing of fluids, in particular liquids, that goes beyond simply passing fluids through a line, in particular a hose or pipe line.

[0092] Advantageously, the bioprocess assembly 1 now advantageously comprises a residence time assembly 28 downstream of the first mixer 27 and fluidly connected to the first mixer 27 to provide a minimum residence time for the liquid streams (see FIG. 2). The third reactive liquid stream 26 is advantageously directed through the residence time assembly 28 to provide a minimum residence time.

[0093] The above-described supply of a liquid stream to the compensation container 2 to maintain a continuous liquid flow results in a change in concentration, advantageously product concentration or buffer concentration, in the compensation container 2. Both parameters can affect the logarithmic reduction of the viral load, so that the residence time of the liquid stream in the residence time assembly 28 is at least 15 minutes, advantageously at most 1 hour, and such changes likewise remain insignificant.

[0094] The term "fluid connection" here means a tight connection that allows fluid to be internally connected from one area to another in at least one direction, preferably in both directions, where the fluid connection can also be released by mechanical release.

[0095] By "residence time assembly" is meant here an assembly that, when properly assembled, is used to cause a volumetric flow of one or more combined fluid streams, particularly liquid streams, to "reside" within the assembly, where the residence time assembly 28 is configured in such a way that the travel distance of the volumetric flow is artificially extended, so that it must travel multiple distances relative to the length of the components. As already explained above, it is important that the distribution of the residence times of the individual volume fractions to be inactivated be as uniform as possible in order to obtain reproducible inactivation results.

[0096] Here, advantageously, the sensor assembly 14 for generating the process data 15 has at least one sensor 29 for measuring a liquid parameter, in particular pH and / or conductivity, of the first liquid stream 24 containing the target factor, which is advantageously configured as a pH sensor and is provided for measuring the pH in the first liquid stream 24 containing the target factor. Alternatively, the sensor 29 is advantageously configured as a conductivity sensor, which is further advantageously provided for measuring the conductivity of the first liquid stream 24 containing the target factor. The sensor 29 is here advantageously arranged downstream of the compensation vessel 2 and upstream of the first bioprocess inlet 22 (FIG. 2).

[0097] Additionally, the sensor assembly 14 here advantageously comprises at least one sensor 30, 31, advantageously at least two sensors 30, 31, for measuring a liquid parameter, in particular pH and / or conductivity, of the third reactive liquid stream 26. The sensor 30 is here advantageously arranged downstream of the first mixer 27. Additionally or alternatively, the sensor assembly 14 here advantageously comprises at least one sensor 31 downstream of the residence time assembly 28 for measuring a liquid parameter in the third reactive liquid stream 26. The sensor 31 is used in particular for measuring flow rate and / or protein concentration, advantageously by UV measurement.

[0098] The measurement of the flow rate is advantageously used for a particularly accurate determination of the required residence time of the third reactive liquid stream 26 in the residence time assembly 28. This is based on the fact, as already discussed above, that a change in the flow rate results in a corresponding change in the residence time of the liquid stream in the residence time assembly 28 that is required for the defined method. The residence time of the liquid stream in the residence time assembly 28 can therefore be set by adapting the flow rate.

[0099] Measurement of UV absorption is advantageously used for particularly accurate determination of target factor concentrations, in particular protein concentrations. In response to a concentration below a predetermined minimum protein concentration, the electronic process control device 7 can control the conveying assembly 6 so that the flow direction of the third reactive liquid stream 26 is diverted via the line 13 of the line assembly 12, which will be described in more detail later, and the third reactive liquid stream 26 can be continuously further conducted to a subsequent process step or discarded in a subsequent discard step. This is advantageously the case when the predetermined minimum protein concentration is below, because otherwise it would not be economical to further inactivate the product. The predetermined minimum protein concentration may be below, in particular, at the beginning (during or immediately after so-called priming) or at the end of a continuous downstream process, in particular a continuous viral inactivation process.

[0100] The electronic process control device 7 advantageously adapts the conveying capacity of the fourth conveying device 23 in the virus inactivation routine 32 to set a target value for the liquid parameter in the third reactive liquid stream 26. This is advantageously done in response to the deviation between an actual value determined by measurement of the liquid parameter of the first liquid stream 24 comprising the target factor and a predefined target value for the liquid parameter in the third reactive liquid stream 26.

[0101] Advantageously, the conductivity enables the electronic process control device 7 to derive the buffer concentration. According to this configuration example, if the actual pH in the first liquid stream 24 comprising the target protein deviates from the setpoint value of the liquid parameter in the third reactive liquid stream 26, the electronic process control device 7 is advantageously able to adapt the actual value of the liquid parameter in the third reactive liquid stream 26, advantageously the actual pH or the actual conductivity, by open-loop and / or closed-loop control of the fourth conveying device 23, which is advantageously provided for the metering delivery of acid, buffer and / or water.

[0102] Thus, by measuring the pH, it is advantageously possible, on the one hand, to determine the actual pH in the first liquid stream 24 containing the target factor, and, on the other hand, the electronic process control device 7 can calculate, based on the process data 15, the volume and / or volume flow rate required to achieve the target value of the liquid parameter, in particular the target pH, in the third reactive liquid stream 26.

[0103] Alternatively, conductivity measurements can advantageously allow, on the one hand, the actual conductivity in the first liquid stream 24, including the target factor, to be determined, and, on the other hand, the electronic process control device 7 can calculate, based on the process data 15, the volume and / or volume flow rate required to achieve the target value of the liquid parameter in the third reactive liquid stream 26, in particular the target conductivity.

[0104] Furthermore, the conduit assembly 12 advantageously includes a waste conduit 33 downstream of the residence time assembly 28 that is provided for conducting the liquid flow from the bioprocess assembly 1 .

[0105] By "waste line" here is meant line 13 of the line assembly 12 for establishing a fluid connection, which offers users of the proposed method the possibility of not necessarily directing a liquid stream introduced into the bioprocess assembly 1 to or from the bioprocess outlet 11. This is advantageous in particular if the bioprocess assembly 1 is washed, for example with a buffer, prior to the proposed continuous downstream process, and in particular if in the proposed first continuous downstream process it is washed with a freshly used buffer and / or a freshly used concentrate stream, in particular a fresh product or buffer concentrate stream.

[0106] Additionally or alternatively, this is the case when the proposed method is used for a continuous in-line dilution process and advantageously the third reactive liquid stream 26 already has the desired predefined target value of the liquid parameter in the third reactive liquid stream 26, thereby representing the final diluted buffer solution. In this case, this final diluted buffer solution is continuously led out of the bioprocess assembly 1 through the waste line 33 and can be directly used in downstream process steps, in particular downstream chromatography steps.

[0107] Here, advantageously, the line assembly 12 is assigned a valve assembly 34 with at least one valve for selective fluid connection of at least one line 13 of the line assembly 12, in particular the waste line 33.

[0108] Advantageously, the bioprocess assembly 1 here advantageously has a second bioprocess inlet 35, advantageously downstream of the waste line 33, provided for introducing a liquid stream into the pipe assembly 12 of the bioprocess assembly 1. The conveying assembly 6 advantageously has a fifth conveying device 36, which is advantageously assigned to the second bioprocess inlet 35 and provided for metering a liquid stream into the pipe assembly 12.

[0109] According to FIG. 2 , the third reactive liquid stream 26 is now advantageously combined downstream of the residence time assembly 28 with the fourth neutralizing liquid stream 37 introduced through the second bioprocess inlet 35 to form a fifth combined liquid stream 38, thereby neutralizing a predefined target value of the liquid parameter in the fifth combined liquid stream 38.

[0110] "Neutralization" here refers specifically to the partial or complete reversal of a previous change in liquid parameters by the reaction of an equal amount of acid, e.g., 1.5-3 M acetic acid, or an acid derivative, e.g., 2 M glycine, and a base, e.g., 1-2 M HEPES pH 8 or Tris pH 11, thereby canceling and / or eliminating the virus inactivation conditions. This fourth neutralized liquid stream 37 is used to neutralize, deplete, and / or eliminate the virus inactivation conditions. Combining the third and fourth liquid streams 26, 37 results in a fifth combined liquid stream 38, which can be discharged from the bioprocess assembly 1 through the bioprocess outlet 11 and has a pH, advantageously between 5 and 8.5, allowing for further processing. Advantageously, the two liquid streams 26, 37 are likewise mixed in a precisely predefined volume ratio, thereby ensuring that the neutralized conditions in the fifth combined liquid stream 38 are actually present and uniformly distributed to perform the intended function.

[0111] Advantageously, in a continuous in-line dilution process, the introduction of the fourth neutralizing liquid stream 37 is optional. If the buffer must be further diluted to achieve the predefined conditions, additional dilution buffer, water, or the like can be added to the third reactive liquid stream 26 through the second bioprocess inlet 35. Alternatively, if the predefined conditions have already been reached, none need be added so that the third reactive liquid stream 26 exits the bioprocess assembly 1 through the bioprocess outlet 11.

[0112] Here, preferably, each conveying device (8, 9, 23, 36), in particular all conveying devices, provided for the metering of the liquid flow, is configured as a metering pump, more preferably as a tube pump, rotary piston pump or diaphragm pump, or as a valve, in particular a continuous valve, a regulating valve or the like.

[0113] The bioprocess assembly 1 here advantageously has at least one second mixer 39 downstream of the second bioprocess inlet 35, which is provided for fluid mixing. The fifth combined liquid stream 38 is advantageously guided through the at least second mixer 39 for fluid mixing and, after fluid mixing, is discharged from the bioprocess assembly 1 through the bioprocess outlet 11. This second mixer 39 may likewise be configured as an in-line static mixer, in particular a radial static mixer or a laminar static mixer, or the like, as shown in FIG. 2. Advantageously, several static mixers may be connected in series, depending on process or precision requirements. Additionally or alternatively, a dynamic mixer may also be provided as the second mixer 39.

[0114] Advantageously, the sensor assembly 14 for generating the process data 15 here comprises at least one sensor 40 for measuring a liquid parameter, in particular pH and / or conductivity, of the fifth combined liquid stream 38. The sensor 40 is here advantageously arranged downstream of the second mixer 39.

[0115] The electronic process control device 7 advantageously adapts the conveying capacity of the fifth conveying device 36 in the neutralization routine 41 to set a target value for the liquid parameter in the fifth combined liquid stream 38. This is advantageously done in response to the deviation between the actual value determined by measurement of the liquid parameter of the third reactive liquid stream 26 and the predefined target value for the liquid parameter of the fifth combined liquid stream 38.

[0116] Advantageously, the conductivity enables the electronic process control device 7 to derive the buffer concentration. According to this configuration example, if the actual pH in the third reactive liquid stream 26 deviates from the target value of the liquid parameter in the fifth combined liquid stream 38, the electronic process control device 7 can advantageously adapt the actual value of the liquid parameter in the fifth combined liquid stream 38, advantageously the actual pH or the actual conductivity, by open-loop and / or closed-loop control of the fifth conveying device 36, which is advantageously provided for the metering of base, buffer and / or water.

[0117] Thus, by measuring the pH, advantageously, on the one hand, the actual pH in the third reactive liquid stream 26 can be determined, and on the other hand, the electronic process control device 7 can calculate, based on the process data 15, the volume and / or volume flow rate required to achieve the target parameters, in particular the target pH, in the fifth combined liquid stream 38.

[0118] Alternatively, conductivity measurements can advantageously allow, on the one hand, the actual conductivity in the third reactive liquid stream 26 to be determined, and, on the other hand, the electronic process control device 7 can calculate, based on the process data 15, the volume and / or volume flow rate required to achieve a target value, in particular a target conductivity, in the fifth combined liquid stream 38.

[0119] According to a particularly preferred configuration, each sensor 16, 29, 30, 31, 40 measures at least one parameter from the group comprising pH value, conductivity, fill level, change in fill level per time, conveying capacity of at least one conveying device, flow rate of at least one liquid stream, protein concentration and / or other spectrophotometric properties of at least one liquid stream to generate the process data 15. In configurations with multiple sensors 16, 29, 30, 31, 40, at least two of them can measure the same or different parameters.

[0120] The electronic process control device 7 can generate a second model 42 from the process data 15. The degree of adaptation of the conveying capacity of the fourth and / or fifth conveying devices 23, 36 is advantageously derived from the second model 42. Advantageously, the second model 42 represents a relationship between the liquid parameters of the third reactive liquid stream 26, on the one hand, and the conveying capacity of the fourth conveying device 23, on the other hand. Additionally or alternatively, the second model 42 advantageously represents a relationship between the fluid parameters of the fifth composite liquid stream 38, on the one hand, and the conveying capacity of the fifth conveying device 36, on the other hand. Based on this second model 42, at least the conveying assembly 6 is advantageously adjusted by the electronic process control device 7. 2, the electronic process controller 7 calculates, based on the process data 15, which volume, volumetric flow rate or the like of the second liquid stream 25 must be added to the first liquid stream 24 containing the target factor in order to produce a target value of the liquid parameter, in particular a target pH or target conductivity, in the third reactive liquid stream 26. Additionally or alternatively, the electronic process controller 7 calculates, based on the process data 15, which volume, volumetric flow rate or the like of the fourth liquid stream 37 must be added to the third reactive liquid stream 26 in order to produce a target value of the liquid parameter, in particular a target pH or target conductivity, in the fifth combined liquid stream 38.

[0121] In particular, in a continuous in-line dilution process, the second model 42 advantageously calculates the degree of adaptation of the delivery capacity of the fourth delivery device 23 based on at least one characteristic curve between the conductivity measured in the first fluid stream 24 containing the target factor and the corresponding buffer concentration. Additionally or alternatively, the second model 42 advantageously calculates the degree of adaptation of the delivery capacity of the fifth delivery device 36 based on at least one characteristic curve between the conductivity measured in the third reactive liquid stream 26 and the corresponding buffer concentration.

[0122] Here, advantageously, the first model 21 and / or the second model 42 are statistical or analytical models.

[0123] The term "statistical model" refers to a mathematical model in which some or all of the input data has some kind of randomness, for example, represented by a probability distribution, so that for a particular set of input data, the output is not reproducible but is described by the probability distribution. The output data is obtained by running the model multiple times with new input values ​​taken from the probability distribution for each run.

[0124] The term "analytical model" refers to a quantitative model used to answer a specific question. Its main purpose is to provide a closed-form expression for a particular property. An analytical model is a mathematical model with a closed-form solution, i.e., the solution to the equations used to describe the evolution of a system can be expressed as a mathematical analytical function.

[0125] Here, advantageously, the liquid streams withdrawn from the bioprocess assembly 1, advantageously the third reactive liquid stream 26 or the fifth combined liquid stream 38, continuously exit the bioprocess assembly 1. Even more advantageously, the withdrawn third reactive liquid stream 26 continuously exits the bioprocess assembly 1 through the waste line 33. Alternatively, the withdrawn fifth combined liquid stream 38 continuously exits the bioprocess assembly 1 through the bioprocess outlet 11.

[0126] By "continuous" here is meant that the liquid flow 25, 35 exiting the bioprocess assembly 1 is not interrupted, for example by incubation in a vessel or the like, but it may vary in flow rate.

[0127] Advantageously, at least the first and / or second mixer 27, 39, and preferably all mixers, are provided for in-line mixing of the liquid streams and are configured as static mixers or as non-positive displacement pumps, in particular centrifugal pumps. It is further advantageous if the non-positive displacement pumps are run in a reverse flow direction compared to the normal operation for dynamic in-line mixing.

[0128] Advantageously, the proposed method can be combined with an upstream, preferably discontinuous or continuous, chromatography method, in particular affinity chromatography and ion exchange chromatography. Additionally or alternatively, the proposed method can be combined with an upstream, preferably discontinuous or continuous filtration method, in particular tangential flow filtration or the like. Additionally or alternatively, the proposed method can be followed downstream by a continuous chromatography or filtration method.

[0129] In principle, the proposed method can be used in combination with all purification, filtration, chromatography, separation, centrifugation, concentration and / or precipitation methods or other methods that can be assigned to downstream processes in bioprocessing.

[0130] According to a further independent teaching, a bioprocess assembly 1 is provided for operating a continuous downstream process during a bioprocess, in particular a continuous viral inactivation process or an in-line dilution process. The bioprocess assembly 1 includes a compensation vessel 2 having a first fluid inlet 3 for introducing a concentrate stream, in particular a product or buffer concentrate stream, into the compensation vessel 2 and at least one second fluid inlet 4 for introducing a diluent stream, in particular a product or buffer diluent stream, into the compensation vessel 2. The bioprocess assembly 1 also includes a fluid outlet 5 for leading a liquid stream from the compensation vessel 2. Furthermore, the bioprocess assembly 1 includes a transport assembly 6 for fluid transport, the transport assembly 6 being assigned to the first fluid inlet 3 and the second fluid inlet 4, respectively, for transporting at least one liquid stream to the compensation vessel 2. Additionally, the transport assembly 6 is assigned to the fluid outlet 5 for transporting the liquid stream from the compensation vessel 2. The bioprocess assembly 1 includes an electronic process control device 7.

[0131] It is important to note here that during the normal operation of the proposed bioprocess assembly 1, the electronic process control device 7 adapts the fill level in the compensation vessel 2 by means of open-loop and / or closed-loop control of the transport assembly 6. This adaptation is performed so as not to fall below a predefined minimum fill level in the compensation vessel 2, thereby ensuring a continuous liquid flow from the bioprocess assembly 1.

[0132] Here, advantageously, at least one component, advantageously all components of the bioprocess assembly 1 are configured as single-use components.

[0133] According to the further teaching of claim 26, which has an independent meaning, patent protection is claimed for the electronic process control device 7 of the proposed bioprocess assembly 1. In this regard, reference may be made to the entire description of the proposed method for operating a continuous downstream process and the proposed bioprocess assembly 1.

[0134] Here, advantageously, the pipeline assembly 12 is assigned a valve assembly 34 with at least one valve for selective fluid connection of at least one pipeline 13, advantageously all pipelines, of the pipeline assembly 12. An electronic process control device 7 is provided for implementing the proposed method by open-loop and / or closed-loop control of at least the transport assembly 6.

[0135] According to a preferred configuration as defined in claim 27, the electronic process control device 7 advantageously comprises a data processing system for implementing the method according to the proposal.

[0136] According to the further teaching of claim 28, which has an independent meaning, patent protection is claimed for the use of the electronic process control device 7 for carrying out the proposed method. In this regard, reference may be made to all descriptions relating to the proposed method for operating a continuous downstream process, the proposed bioprocess assembly 1, and the proposed electronic process control device 7.

[0137] According to the further teaching of claim 29, which has an independent meaning, patent protection is claimed for a computer program for the proposed electronic process control device 7. In this regard, reference may be made to all descriptions relating to the proposed method for operating a continuous downstream process, the proposed bioprocess assembly 1, the proposed electronic process control device 7 and the proposed use.

[0138] According to the further teaching of claim 30, which has an independent meaning, patent protection is claimed for a computer-readable storage medium for storing the proposed computer program. In this regard, reference may be made to all descriptions relating to the proposed method for operating a continuous downstream process, the proposed bioprocess assembly 1, the proposed electronic process control device 7, the proposed use and the proposed computer program.

Claims

1. A method for operating a continuous downstream process, in particular a continuous viral inactivation process or an in-line dilution process, during a bioprocess using a bioprocess assembly (1), comprising a compensating vessel (2) having a first fluid inlet (3) for introducing a concentrate stream, in particular a product or buffer concentrate stream, into said compensating vessel (2), at least one second fluid inlet (4) for introducing a diluent stream, in particular a product or buffer diluent stream, into said compensating vessel (2), and a compensation vessel (2) having a fluid outlet (5) provided for leading a liquid flow from the compensation vessel (2), the bioprocess assembly (1) having a transport assembly (6) for fluid transport, the transport assembly (6) being assigned to the first fluid inlet (3) and the second fluid inlet (4), respectively, for transporting at least one liquid flow to the compensation vessel (2) and the fluid outlet (5) for transporting a liquid flow from the compensation vessel (2), the bioprocess assembly (1) having an electronic process control device (7), The method is characterized in that the electronic process control device (7) adapts the filling level in the compensation container (2) by open-loop and / or closed-loop control of the transport assembly (6) so that it does not fall below a predefined minimum filling level in the compensation container (2).

2. 2. The method according to claim 1, characterized in that the electronic process control device (7) adapts the filling level in the compensation container (2) by open-loop and / or closed-loop control of the conveying assembly (6) so as to achieve a predefined target filling level or to maintain the filling level within a target filling level range.

3. 3. The method according to claim 1 or 2, characterized in that the conveying assembly (6) has a first conveying device (8) for fluid conveyance assigned to the first fluid inlet (3), and / or the conveying assembly (6) has a second conveying device (9) for fluid conveyance assigned to the second fluid inlet (4), and / or the conveying assembly (6) has a third conveying device (10) for fluid conveyance assigned to the fluid outlet (5).

4. 4. The method according to claim 1, wherein the bioprocess assembly (1) comprises a bioprocess outlet (11) provided for leading a fluid out of the bioprocess assembly (1), the bioprocess assembly (1) comprises a conduit assembly (12) with at least one conduit (13) for fluidly connecting the fluid outlet (5) of the compensation vessel (2) to the bioprocess outlet (11), and the third conveying device (10) is provided for conveying a fluid through the at least one conduit (13) of the conduit assembly (12).

5. The bioprocess assembly (1) comprises a sensor assembly (14) for generating process data (15) comprising at least one sensor (16) assigned to the compensation vessel (2), and the electronic process control device (7) derives, in a monitoring routine (17), from the process data (15) an actual value of a fill level parameter in the compensation vessel (2); and adapting the conveying capacity of the first, second and / or third conveying devices (8, 9, 10) in response to a deviation between the derived actual value of the filling level parameter and a predefined minimum value of the filling level parameter corresponding to the predefined minimum filling level in the compensation container (2) in order to prevent the minimum filling level from being exceeded, or adapting the conveying capacity of the first, second and / or third conveying devices (8, 9, 10) in dependence on the deviation between the derived actual value of the filling level parameter and a predefined target value of the filling level parameter, which corresponds to a predefined target filling level in the compensation container (2), or Adapting the conveying capacity of the first, second and / or third conveying devices (8, 9, 10) in response to a deviation between the derived actual value of the filling level parameter and a predetermined limit value of a predetermined target filling level range in the compensation container (2) in order to maintain the filling level within the target filling level range.

5. The method according to claim 1, wherein the first and second electrodes are connected to each other.

6. 6. The method according to claim 1, wherein the electronic process control device (7) generates a first model (21) from the process data (15), and the degree of adaptation of the conveying capacity of the conveying assembly (6) is derived from the first model (21), preferably the first model (21) representing a relationship between, on the one hand, a fill level parameter in the compensation container (2) and, on the other hand, the conveying capacities of the first, second and / or third conveying devices (8, 9, 10).

7. 7. The method according to claim 1, wherein the bioprocess assembly (1) comprises a first bioprocess inlet (22) downstream of the compensation vessel (2) for introducing a liquid flow into the pipeline assembly (12) of the bioprocess assembly (1), and the conveying assembly (6) comprises a fourth conveying device (23) assigned to the first bioprocess inlet (22) for metering a liquid flow into the pipeline assembly (12).

8. 8. The method according to claim 1, wherein the concentrate stream is combined with the diluate stream in the compensation vessel (2) to form a first liquid stream (24) comprising a target factor, and the first liquid stream (24) comprising the target factor is introduced from the compensation vessel (2) through the fluid outlet (5) into at least one fluidly connected pipe (13) of the pipe assembly (12), and is combined in a predefined volume ratio with a second liquid stream (25) introduced preferably through a first bioprocess inlet (22) and conveyed by a fourth conveying device (23) to form a third reactive liquid stream (26), thereby setting predefined target values ​​of liquid parameters, in particular pH and / or conductivity, in the third reactive liquid stream (26).

9. 9. The method according to claim 8, characterized in that the bioprocess assembly (1) advantageously comprises a first mixer (27) provided for fluid mixing downstream of the first bioprocess inlet (22), and the third reactive liquid stream (26) is led through the first mixer (27) for fluid mixing.

10. 10. The method according to claim 8 or 9, characterized in that the bioprocess assembly (1) advantageously comprises, downstream of the at least one mixer (27), a residence time assembly (28) fluidly connected to a first of the mixers (27) to provide a minimum residence time for the liquid flows, the third reactive liquid flow (26) being led through the residence time assembly (28) subsequent to the at least one mixer (27) to provide a minimum residence time.

11. 11. The method according to claim 7, wherein the sensor assembly (14) for generating the process data (15) comprises at least one sensor (29) for measuring a liquid parameter, in particular pH and / or conductivity, of the first liquid stream (24) comprising the target factor, and at least one sensor (30, 31), preferably at least two sensors (30, 31), for measuring a liquid parameter, in particular pH and / or conductivity, of the third reactive liquid stream (26), preferably arranged downstream of the first mixer (27) and / or the residence time assembly (28).

12. 12. The method according to claim 11, characterized in that the electronic process control device (7) adapts the conveying capacity of the fourth conveying device (23) to set the target value of the liquid parameter in the third reactive liquid stream (26) in the virus inactivation routine (32) depending on the deviation between the actual value determined by measuring the liquid parameter of the first liquid stream (24) containing the target factor and a predefined target value of the liquid parameter in the third reactive liquid stream (26).

13. 13. The method according to any one of claims 1 to 12, characterized in that the conduit assembly (12) has a waste conduit (33), preferably downstream of the residence time assembly (28), provided for conducting the liquid flow from the bioprocess assembly (1).

14. 14. The method according to claim 1, wherein the line assembly (12) is assigned a valve assembly (34) with at least one valve for selective fluid connection of at least one line (13) of the line assembly (12), in particular a waste line (33).

15. 15. The method according to claim 1, wherein the bioprocess assembly (1) has a second bioprocess inlet (35), preferably downstream of the waste line (33), provided for introducing a liquid stream into the pipeline assembly (12) of the bioprocess assembly (1), and the conveying assembly (6) has a fifth conveying device (36), assigned to the second bioprocess inlet (35), provided for metering a liquid stream into the pipeline assembly (12), and wherein the third reactive liquid stream (26) is preferably combined downstream of the residence time assembly (28) with a fourth neutralizing liquid stream (37), introduced through the second bioprocess inlet (35), to form a fifth combined liquid stream (38), thereby neutralizing predefined liquid parameters in the fifth combined liquid stream (38).

16. 16. The method according to claim 1, wherein each conveying device (8, 9, 23, 36), preferably all conveying devices provided for metering the liquid flow, are configured as metering pumps, in particular tube pumps, rotary piston pumps or diaphragm pumps, or as valves, in particular continuous valves, control valves or the like.

17. 17. The method according to any one of claims 1 to 16, characterized in that the bioprocess assembly (1) advantageously comprises, downstream of the second bioprocess inlet (35), at least one second mixer (39) provided for fluid mixing, and the fifth combined liquid stream (38) is led through said second mixer (39) for fluid mixing.

18. 18. The method according to claim 1, wherein the sensor assembly (14) for generating the process data (15) preferably comprises at least one sensor (40) downstream of the second mixer (39) for measuring a liquid parameter of the fifth combined liquid stream (38), in particular pH and / or conductivity, and wherein the electronic process control device (7) adapts the conveying capacity of the fifth conveying device (36) in the neutralization routine (41) in dependence on the deviation between an actual value determined by measuring the liquid parameter of the third reactive liquid stream (26) and a predefined target value of the liquid parameter of the fifth combined liquid stream (38) to set the target value of the liquid parameter of the fifth combined liquid stream (38).

19. 19. The method according to claim 1, wherein each sensor (16, 29, 30, 31, 40) for generating the process data (15) measures at least one parameter from the group comprising pH value, conductivity, fill level, change in fill level per time, conveying capacity of at least one conveying device, flow rate of at least one liquid stream, protein concentration and / or other spectrophotometric properties of at least one liquid stream.

20. 20. The method according to claim 1, wherein the electronic process control device (7) generates a second model (42) from the process data (15), and the degree of adaptation of the conveying capacity of the fourth and / or fifth conveying devices (23, 36) is derived from the second model (42), preferably the second model (42) representing the relationship between the liquid parameters of the third reactive liquid stream (26) on the one hand and the conveying capacity of the fourth conveying device (23) on the other hand, and / or the second model (42) representing the relationship between the liquid parameters of the fifth composite liquid stream (38) on the one hand and the conveying capacity of the fifth conveying device (36) on the other hand.

21. Method according to any one of claims 6 to 20, characterized in that the first model (21) and / or the second model (42) are statistical or analytical models.

22. 22. The method according to any one of claims 1 to 21, characterized in that the liquid streams withdrawn from the bioprocess assembly (1), preferably the third reactive liquid stream (26) or the fifth combined liquid stream (38), leave the bioprocess assembly (1) continuously.

23. 23. The method according to claim 1, wherein at least the first and / or second mixer (27, 39), preferably all mixers, are provided for in-line mixing of the liquid streams and are configured as static mixers or as non-positive displacement pumps, in particular centrifugal pumps, and more preferably the non-positive displacement pumps are run through in a reverse flow direction compared to normal operation for dynamic in-line mixing.

24. 24. The method according to any one of claims 1 to 23, characterized in that the proposed method is carried out in combination with an upstream, preferably discontinuous or continuous, chromatographic and / or filtration method and / or the proposed method is followed downstream by a continuous chromatographic or filtration method.

25. A bioprocess assembly (1) for operating a continuous downstream process, in particular a continuous virus inactivation process or an in-line dilution process, during a bioprocess, said bioprocess assembly (1) comprising a compensating vessel (2) having a first fluid inlet (3) for introducing a concentrate stream, in particular a product or buffer concentrate stream, into said compensating vessel (2), at least one second fluid inlet (4) for introducing a diluent stream, in particular a product or buffer diluent stream, into said compensating vessel (2), and at least one third fluid inlet (5) for withdrawing a liquid stream from said compensating vessel (2). a compensation vessel (2) having a fluid outlet (5) for transferring a liquid stream from the compensation vessel (2), the bioprocess assembly (1) having a transport assembly (6) for transferring fluids, the transport assembly (6) being assigned to the first fluid inlet (3) and the second fluid inlet (4), respectively, for transferring at least one liquid stream to the compensation vessel (2), and the fluid outlet (5) for transferring a liquid stream from the compensation vessel (2), the bioprocess assembly (1) having an electronic process control device (7), During normal operation of the bioprocess assembly (1), the electronic process control device (7) adapts the filling level in the compensation container (2) by open-loop and / or closed-loop control of the transport assembly (6) so that it does not fall below a predefined minimum filling level in the compensation container (2).

26. 26. An electronic process control device (7) for a bioprocess assembly (1) according to claim 25, wherein a pipeline assembly (12) is assigned a valve assembly (34) with at least one valve for selective fluid connection of at least one pipeline (13), preferably all pipelines, of the pipeline assembly (12), the electronic process control device (7) controls at least the transport assembly (6) or additionally the valve assembly (34), and the electronic process control device (7) is provided for carrying out the method according to any one of claims 1 to 24 by open-loop and / or closed-loop control of at least the transport assembly (6).

27. 27. An electronic process control device (7) according to claim 26, characterized in that the electronic process control device (7) comprises a data processing system for implementing the method according to any one of claims 1 to 24.

28. 28. Use of an electronic process control device (7) according to claim 26 or 27 for carrying out a continuous downstream process according to any one of claims 1 to 24.

29. 28. Computer program for an electronic process control device (7) according to claim 26 or 27.

30. 30. A computer-readable storage medium on which the computer program according to claim 29 is stored, preferably non-volatilely.

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