Method for operating the clarification setup

Abstract

The present invention is directed to a method for operating a clarification setup (1) of a bioprocessing installation (2), the clarification setup (1) comprising a fluidized bed centrifuge (3) for clarification of a cell broth by centrifugation, the fluidized bed centrifuge (3) comprising at least two centrifuge chambers (9-12), the fluidized bed centrifuge (3) being operated in a forward operation for a particle loading cycle and / or a particle washing cycle and in a reverse operation for a particle discharge cycle, during which the cell broth loaded in the centrifuge chambers (9-12) progresses to form a growing particle accumulation in the centrifuge chambers (9-12), the clarification setup (1) comprising a monitoring sensor device (17) with at least one, preferably optical, sensor (18) for generating monitoring sensor data that are transmitted to an electronic process control device (14). It is proposed that in a monitoring routine, for at least some of the centrifuge chambers (9-12) monitoring sensor data (19) are generated by the monitoring sensor device (17) individually, and that during a loading cycle, in an adjustment routine at least one parameter of the clarification setup (1) is adjusted based on the monitoring sensor data (19) so as to control the loading of cell broth individually for at least some of the centrifuge chambers (9-12) according to a predefined adjustment strategy.

Description

[Technical field]

[0001] This application has been filed as a PCT international patent application dated January 11, 2023 in the name of Sartorius Stedim North America Inc., a U.S. corporation, applicant in all countries, and Robert Soeldner, a German national, Jonas Austerjost, a German national, and Martin Saballus, a German national, inventors and applicants in all countries. This application claims priority to EP Application No. 22151043.1, filed January 11, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] The present invention relates to a method for operating a clarification setup of a bioprocessing installation according to the general part of claim 1, a clarification setup according to claim 13 and an electronic process control device according to claim 14. [Background technology]

[0003] The term "bioprocess" currently refers to any kind of biotechnological process, especially biopharmaceutical processes. An example of such a bioprocess is the use of a bioreactor to cultivate microorganisms or mammalian cells under given conditions, where the cell broth is transferred from the bioreactor to a downstream process. The term "bioreactor" in this context means any manufactured device or system that supports a biological environment by allowing the monitoring and control of at least one parameter. The term "biological product" currently refers to any kind of compound produced in such a bioprocess. Examples of such biological products are proteins, especially antibodies, growth factors or hormones, metabolites or any other molecules, as well as cells or cell components, e.g. organelles.

[0004] The method for operating a clarification setup can be applied in various fields of biotechnology. High efficiency in this field has been driven by the increasing demand for biological products such as biopharmaceuticals. Efficiency in this sense is not only related to the cost-effectiveness of the components used but also to the controllability of the processes associated with it. The method relies on optimal filling of the centrifuge chambers to optimize the time efficiency as well as the controllability of the bioprocess.

[0005] A known method for operating a clarification setup (EP 2310486) comprises a fluidized bed centrifuge with a number of centrifuge chambers, each of which is assigned a chamber inlet and a chamber outlet. The clarification setup also comprises a pumping device assigned to the fluidized bed centrifuge and a liquid network for transporting the liquid, which is the cell broth to be clarified. Finally, the clarification setup comprises an electronic process control device for controlling at least the fluidized bed centrifuge and the pumping device.

[0006] Known clarification setups with one or more centrifuge chambers generally allow high output performance since the centrifugation processes of each centrifuge chamber are performed in parallel. However, performing the various centrifugation processes in parallel is complicated if maximum efficiency is intended, since the centrifugation processes of each centrifuge chamber must be considered as separate processes with their own dynamics.

[0007] Another known method, which is the starting point of the present invention ("Operation and Maintenance Manual kSep400", Sartorius Stedim Biotech), relies on the generation of monitoring sensor data from a camera sensor, which is manually analyzed by a user. Based on the user's experience, the monitoring sensor data are utilized to operate the clarification setup. Although the use of a camera sensor facilitates the analysis by the user, maximizing the efficiency of the clarification setup remains a challenge due to the reasons mentioned above. Summary of the Invention [Problem to be solved by the invention]

[0008] It is therefore an object of the present invention to improve the known methods for operating a clarification set-up in such a way that the bioprocess efficiency is improved with as little effort as possible. [Means for solving the problem]

[0009] The above mentioned problem is solved in a method for operating a clarification setup as set out in the general part of claim 1 by the features of the characterizing part of claim 1.

[0010] The basic idea underlying the present invention is to individually control the loading of cell broth in at least some of the centrifuge chambers based on monitoring sensor data which may, for example, represent the individual particle loading level of each centrifuge chamber.

[0011] The term "particle" should be understood in the broad sense and means any particulate solid compound which may be of biological or non-biological origin, such as in particular cells, cell debris, proteins, (magnetic) microbeads, etc.

[0012] The term "particle filling level" should be understood in a broad sense. Particle filling level is a measure of the filling of the centrifuge chamber and can be reflected by the particle filling rate of the centrifuge chamber, a specific particle mass, volume, concentration, etc. Furthermore, particle filling level can be expressed by a specific particle filling level at a given time or as a change in particle filling level over time, which is also reflected in the particle filling rate.

[0013] The term "individually" here generally means that the respective task, here the control of the loading, is performed for each centrifuge chamber individually. In particular, the synchronization of the loading of the centrifuge chambers can be performed in various ways according to a predefined coordination strategy.

[0014] Depending on the regulation strategy, it may be possible, for example, to allow uniform filling of each centrifuge chamber, which allows simultaneous emptying of all centrifuge chambers and therefore more efficient use of the centrifuge chamber capacity with less effort.

[0015] Synchronized loading of the centrifuge chambers can reduce unnecessary waiting times, minimize the number of centrifugation cycles required to process a bioreactor volume, and prevent particle breakthrough of valuable cells. The term "particle breakthrough" refers to the point in time during a fluidized bed centrifugation process when at least one centrifuge chamber is filled to capacity with particles, preferably cells, and continued loading of the centrifuge chambers leads to overfilling of said centrifuge chamber.

[0016] The proposed individual control of the loading of each centrifuge chamber can also be advantageous with a view to a mechanically balanced operation of the rotor of the fluidized bed centrifuge supporting the centrifuge chambers. Unbalance occurs when the axis of inertia of the rotor relative to the centrifuge axis is offset with respect to the centrifuge axis. This can be caused, for example, by uneven loading of the centrifuge chambers with particles, such as cells, during the loading cycle. By a specific synchronized loading and thus a predefined distribution of the weight of the particles over the centrifuge chambers, a balanced operation can be achieved or at least supported.

[0017] In particular, it is proposed that in a monitoring routine, for at least some of the centrifuge chambers, monitoring sensor data are generated by the monitoring sensor device individually, and during a loading cycle, in an adjustment routine, at least one parameter of the clarification setup is adjusted by the electronic process control device based on the monitoring sensor data so as to control the loading of cell broth individually for at least some of the centrifuge chambers according to a predefined adjustment strategy. Here again, the term "individual" means that each centrifuge chamber is treated individually here, both in terms of generating the monitoring sensor data and in terms of loading of cell broth.

[0018] According to a preferred embodiment, it was found that particle accumulation proceeds in an orderly and reproducible manner due to the suspended state of the particles in the centrifuge chamber. To a large extent, this effect occurs independently of the medium being treated. In particular, it was found that this results in a visible phase boundary, which moves across the centrifuge chamber with the ongoing particle accumulation, the position of the phase boundary being representative of the target filling level of the centrifuge chamber. It is particularly interesting that, in contrast to known non-fluidized bed centrifuges, the proposed invention has been realized to result in a visible phase boundary without subjecting the centrifuged cells to such high acceleration forces that could otherwise lead to cell damage and even cell death. Thus, the cells are separated here under milder conditions, leading to a higher viability, which is crucial if the cells are to be reused in a subsequent bioprocess. Furthermore, the proposed invention allows a more accurate estimation of the number of cells in the centrifuge chamber. This is due to the fact that particle accumulation under non-fluidized bed conditions is fundamentally different due to the much higher acceleration forces acting on the cells. For this reason, in the proposed solution, the cells of the particle accumulation are not packed as tightly as they would be under non-fluidized bed conditions, which allows a more accurate calculation of the particle loading level, which is particularly advantageous for a more accurate calculation of the required number of cells required to inoculate a potential subsequent bioprocess. Furthermore, it has been found that the above-mentioned systematic particle accumulation allows a systematic calculation of the loading level based on a computational model, which can also be performed in real time.

[0019] Therefore, according to claim 2, it is proposed in particular in the monitoring routine that image-related data representative of an optical image of the centrifuge chamber contents is generated by the monitoring sensor device, which further preferably forms the basis for the calculation of the particle filling level of the respective centrifuge chamber according to claim 3.

[0020] By using automatic filling level determination by image processing, an accurate signal for each chamber can be obtained. As an image-related approach, the determined filling level is independent of broth composition such as cell viability, cell type and cell diameter, and can even reflect the presence of impurities or dead cells.

[0021] The individual control of the loading of at least some of the centrifuge chambers is preferably realized by individually controlling at least some of the pumps of the pumping device according to claim 4. Preferably, the pump speed of each of those pumps is then controlled according to a regulation strategy. This approach to controlling the loading of the cell broth is easy to realize from an engineering point of view.

[0022] Claims 5 to 10 are directed to a preferred regulation strategy based on specific control of the individual particle filling level of each centrifuge chamber. This direct control of the individual particle filling level of each centrifuge chamber leads to a simple way of increasing the effectiveness, as proposed in claims 6 to 10. Whereas claims 6 and 7 propose to reach a target, e.g. a target particle filling level, for each centrifuge chamber individually, claims 8 to 10 focus on controlling the particle filling levels of at least some of the centrifuge chambers relative to each other.

[0023] A preferred means for achieving the above mentioned mechanical balancing of the rotor of a centrifuge is the subject of claim 11, which takes into account a balancing model describing the interrelationship between the difference between the particle filling levels of at least two centrifuge chambers and the rotor imbalance, thereby making it possible to prevent rotor imbalance conditions from occurring in the first place.

[0024] As an alternative, claim 12 proposes taking into account the sensor signal of a vibration sensor in order to minimize the amplitude of those vibrations caused by an unbalanced operation of the rotor of the centrifuge. This is done by a method for controlling the volumetric flow rate of at least a part of a pump assigned to a centrifuge chamber taking into account the detected vibrations.

[0025] A clarification set-up for carrying out the proposed method is so claimed according to the teaching of the second independent claim according to claim 13. All explanations given for the proposed method are fully applicable to the proposed clarification set-up.

[0026] The teaching of the third independent claim according to claim 14 is directed to an electronic process control device, which is designed to implement the proposed method according to any one of the previous claims. Here again, all the explanations given above apply in their entirety to this proposed third teaching. The electronic process control device preferably includes a data processing system for implementing the proposed method.

[0027] A fourth teaching, which may also be stated as an independent claim, is directed to a computer program product for the proposed electronic process control device, which is configured to implement the proposed method, in particular the above-mentioned routines. Again, all explanations given for the proposed method are fully applicable to the proposed computer program product.

[0028] A fifth teaching, which may also be written as an independent claim, is directed to a computer-readable storage medium on which a computer program is stored. Here too, all the explanations given for the proposed method are fully applicable to the proposed readable storage medium.

[0029] In the following, one embodiment of the present invention will be described with reference to the drawings. [Brief description of the drawings]

[0030] [Figure 1] FIG. 1 is a schematic diagram of a preferred embodiment of the proposed bioprocessing facility in which the proposed method can be performed. [Diagram 2] FIG. 2 is a perspective view of the operating principle of the method proposed according to FIG. 1. [Diagram 3] FIG. 2 shows the individual filling levels of the centrifuge chambers of the centrifuge shown in FIG. 1 during a loading cycle, a) without application of the proposed adjustment routine, b) with application of the proposed adjustment routine. [Figure 4] FIG. 2 shows a centrifuge and a pumping device according to FIG. 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] The proposed method for operating the clarification setup 1 of a bioprocessing installation 2 is preferably assigned to the upstream and downstream processes of a bioprocess treating liquids, in particular cell broths for cell culture and / or bioproduction.

[0032] The term "liquid" should be understood in a broad sense: it includes not only pure liquids but also emulsions and suspensions, e.g. heterogeneous mixtures of at least two different liquids or heterogeneous mixtures of solid particles and liquids.

[0033] The term "cell broth" refers to a suspension of particles in a solvent, specifically cells and / or cell debris in a medium. It specifically describes the entire culture medium and the respective organisms cultured in the culture medium.

[0034] The term "upstream processing" includes all steps related to cell banking, inoculum (seed train) development, medium development, growth rate optimization and the cultivation process itself, as well as the corresponding in-process control. Cell harvesting can be considered as both part of upstream processing and part of downstream processing. The term "downstream processing" includes all steps related to the recovery and purification of biological products, especially biopharmaceuticals, from natural sources such as animal or plant tissues or cell broths, including recycling of recoverable components and proper treatment and disposal of waste products.

[0035] Generally, cell culture is currently used to produce biopharmaceuticals, especially proteins, such as human insulin, growth factors, hormones, vaccines, or antibodies, antibody derivatives, etc. The biological product may also be a non-biological drug, such as enzymes for food processing, enzymes for laundry detergent, biodegradable plastics, or biofuels. The focus of the present invention is the biopharmaceutical products secreted from cells into the supernatant, such as antibodies or exosomes. Additionally or alternatively, the product may be the cells themselves, especially mammalian cells, including stem cells, or immune cells, such as CAR-T cells for cancer treatment.

[0036] As shown in Figures 1-4, according to all embodiments, the proposed method for operating a clarification setup 1 of a bioprocessing installation 2 employs at least one fluidized bed centrifuge 3 for clarification of cell broth by centrifugation and a pumping device 4 assigned to the fluidized bed centrifuge 3. The pumping device 4 comprises at least two pumps 5-8, here preferably four pumps 5-8. The fluidized bed centrifuge 3 comprises at least one centrifuge chamber 9-12, preferably an even number of centrifuge chambers 9-12, more preferably just four centrifuge chambers 9-12, which are rotated around a geometric centrifuge axis 13, giving rise to a fluidized bed during normal operation. As shown in Figure 4, preferably each centrifuge chamber 9-12 comprises at least one assigned pump 5-8, here preferably one single pump 5-8, for pumping liquid into and out of the chambers 9-12. The bioprocessing installation 2 further comprises an electronic process control device 14 for controlling at least the fluidized bed centrifuge 3 and the pumping device 4.

[0037] "Centrifugation" is the term used to describe the sedimentation of particles in an artificially created gravitational field, where large acceleration forces are used to achieve significant reductions in separation times.

[0038] Here, the centrifuge is designed as a fluidized bed centrifuge 3 for carrying out a continuous centrifugation process. A preferred set-up of a fluidized bed centrifuge 3 is described in EP 2 485 846 A1, the contents of which are incorporated herein by reference.

[0039] The fluidized bed centrifuge 3 comprises a rotor 15 with centrifuge chambers 9-12 mounted within a centrifuge housing 16. The rotor 15 can be rotated about a single geometric centrifuge axis 13, preferably by an electric motor. The term "single" means that all chambers 9-12 supported by the rotor 15 rotate about one and the same geometric centrifuge axis 13. The centrifuge rotation speed and pump speed are adjustable by an electronic process controller 14 with the aim of establishing a fluidized bed of particles such as cells or cell debris in the fluidized bed centrifuge 3. A fluidized bed is achieved when the centrifugal force on the particles is equal to the force of the opposing fluid flow, resulting in zero net force on the particles.

[0040] According to Figure 1, the cell broth is led to a fluidized bed centrifuge 3. The fluidized bed centrifuge 3 is operated in forward operation for the loading and / or washing cycles and in reverse operation for the particle discharge cycle.

[0041] "Forward operation" refers to one of the two possible fluid flow directions in a fluidized bed centrifuge and describes an operation that leads to the separation of liquid, such as culture medium and cells, from solid particles. This "forward operation" allows, on the one hand, washing the separated cells with a buffer or medium, preferably a culture medium, more preferably a concentrated medium, and / or, on the other hand, clarification of the cell broth. The goal here is to clarify the liquid supernatant from solid particles, such as cells, cell debris, etc., which are considered as biomass. The product obtained in this forward operation is the supernatant of the cell broth, which contains the desired bioproduct, such as a recombinant protein, in particular an antibody.

[0042] The term "enriched medium" describes a medium that contains higher concentrations of vitamins, growth factors, micronutrients, and carbon sources, nitrogen sources and / or amino acid concentrations, etc., and preferably allows each organism to grow at its maximum growth rate due to optimized nutrient concentrations. Growth factors and micronutrients are included in the medium for organisms that cannot produce all the necessary vitamins themselves. Inorganic nutrients, including trace elements such as iron, zinc, copper, manganese, molybdenum and cobalt, are typically included in unrefined carbon and nitrogen sources, but may have to be added if refined carbon and nitrogen sources are used.

[0043] The loading cycle refers to the cycle of loading the cell broth to be centrifuged into each centrifuge chamber 9-12 in the forward operation of the fluidized bed centrifuge 3. Thus, during the particle loading cycle, the cell broth loaded into the centrifuge chambers 9-12 proceeds to form a growing particle accumulation in the centrifuge chambers 9-12.

[0044] A wash cycle refers to a cycle in which each of the centrifuge chambers 9-12 is washed with culture medium or buffer during forward operation of the fluidized bed centrifuge 3. This wash cycle preferably serves to supply fresh nutrients to the cells.

[0045] Alternatively, the fluidized bed centrifuge 3 can be operated in reverse operation. "Reverse operation" means the second of the two possible fluid flow directions in the fluidized bed centrifuge 3 and describes an operation that leads to the discharge of separated solid particles, preferably cells. The resulting product in reverse operation is cells in a cell broth.

[0046] The discharge cycle therefore refers to the cycle assigned to the fluidized bed centrifuge 3, during which, in reverse operation, solid particles, preferably cells, are discharged from the centrifuge chambers 9-12. This discharge cycle serves, inter alia, for the reuse of cells in a subsequent bioprocess.

[0047] Furthermore, the clarification setup 1 includes a monitoring sensor device 17 with at least one, here preferably optical, sensor 18, generating monitoring sensor data which are transmitted to the electronic process control device 14. The optical sensors 18 are directed towards each of the centrifuge chambers 9-12 as shown in FIG.

[0048] Of particular importance to the invention is that during the monitoring routine, monitoring sensor data 19 are generated individually for at least some of the centrifuge chambers 9-12 by the monitoring sensor device 17. From these monitoring sensor data 19, the electronic process control device 14 preferably calculates the particle filling level 20 on the basis of a computational model, as will be explained later.

[0049] Furthermore, what is important for the present invention is that during the loading cycle, in an adjustment routine, at least one parameter of the clarification setup 1 is adjusted by the electronic process control device 14 based on the monitoring sensor data 19 in order to control the loading of cell broth for at least some of the centrifuge chambers 9-12, here preferably for all centrifuge chambers, individually according to a predefined adjustment strategy. This means that the monitoring sensor data generated for the individual centrifuge chambers 9-12 are the basis for controlling the loading of cell broth for those individual centrifuge chambers 9-12. This allows an almost unlimited flexibility of individual control of the loading of each and any centrifuge chamber 9-12 according to a predefined adjustment strategy.

[0050] As shown in FIG. 2, during the monitoring routine, the monitoring sensor device 17 generates image related data 21 as monitoring sensor data 19, which represents optical images of each of the centrifuge chamber contents 23-26.

[0051] Preferably, the monitored sensor data 19 is representative of a particle filling level 20 of the respective centrifuge chamber 9-12. This particle filling level 20 may also be derived from the monitored sensor data 19, preferably by calculation based on a computational model.

[0052] The computational model is to be understood as a rule system for calculating the filling level 20 based on the image related data 21. In a preferred embodiment, the computational model can be exchanged between two different operation instances or even within one and the same operation instance. Furthermore, the computational model is highly adaptable to different bioprocess settings resulting in altered optical properties, such as those concerning the choice of culture medium used, which may result in different contrast, different brightness, different density, etc. According to another preferred embodiment, the computational model can be adapted to different cell types, including different particle sizes, different shapes, different concentrations, etc. This adaptability makes the proposed method very flexible.

[0053] The term "image related data" should be understood in a broad sense. It represents at least one image and in this sense may be a regular photographic representation. Furthermore, the term "image related data" may include monitoring sensor data of other sensors of the monitoring sensor device 17 relating to other properties of the liquid. Such properties may be, for example, flow weight, flow rate, cell type, carbon source concentration, nitrogen source concentration, amino acid concentration, pH, temperature, oxygen concentration, carbon dioxide concentration, electrical conductivity, pressure, DNA concentration, protein concentration or biomass concentration.

[0054] In the embodiment according to Fig. 2, here preferably in the monitoring routine, the particle filling level 20 has been calculated according to a calculation model on the basis of different optical properties of the centrifuge chamber contents 23-26. These different optical properties are in particular different translucency, different colour and / or different brightness of the centrifuge chamber contents 23-26 inside and outside the particle accumulation. This may be for example a different colour of solid particles, preferably cells, caused by properties specific for a certain particle type, preferably cell type, such as different turbidity, colour-brightness and / or density difference inside and outside the particle accumulation. The term "particle accumulation" here means the mass of particles accumulated in the centrifuge chambers 9-12.

[0055] In a preferred embodiment, in the monitoring routine, a phase boundary 27 between the particle accumulation of the centrifuge chamber contents 23-26 and the particle-free remaining part is detected in the optical image 22 according to a computational model. Such a phase boundary 27 may arise due to differences in composition between the particle accumulation of the centrifuge chamber contents 23-26 and the particle-free remaining part, such as differences in density, differences in affinity, e.g. differences in hydrophilic or hydrophobic properties, differences in color, translucency and / or brightness. Then, based on the computational model, the particle filling level 20 is calculated from the position of the phase boundary 27. The phase boundary 27 can be detected by image processing, e.g. line recognition algorithms. It can also be detected simply by determining two regions with different optical properties, such as contrast, brightness, etc., in the optical image 22 representing the centrifuge chamber contents 23-26. These different optical properties preferably represent the particle accumulation (e.g. comprising a lower brightness) on the one hand and the particle-free remaining part (e.g. comprising a higher brightness) of the centrifuge chamber contents 23-26 on the other hand.

[0056] Preferably, as seen in Figures 1 and 2, the at least one optical sensor 18 of the monitoring sensor device 17 is a camera unit, preferably a 2D or 3D camera. Exemplary 2D cameras used are two-dimensional CCD array sensors used in video cameras and digital cameras and CMOS sensors used in smartphones and tablets. The optical sensor 18 is preferably located inside or outside the fluidized bed centrifuge 3, more preferably inside or outside the centrifuge rotor chamber.

[0057] The viewing direction of the optical sensor 18, here preferably a camera unit, is preferably essentially parallel to the geometrical centrifuge axis 13. However, it may also be inclined, which may be advantageous in terms of optimized use of the available space. The term "essentially parallel" here does not mean an ideal parallel line in the mathematical sense, but in the colloquial sense that the camera unit is arranged in a better part parallel to the geometrical centrifuge axis 13 (see FIG. 2).

[0058] In Fig. 2, two options for the light device 28, 29 are shown. In the first option, the light of the light device 29 is transmitted through the centrifuge chamber contents 23-26 and directed to the monitoring sensor device 17. This improves the reproducibility of the measurements by the optical sensor 18, independent of any other ambient conditions. In an alternative embodiment included in Fig. 2, the light of the light device 28 is directed to the centrifuge chamber contents 23-26 and reflected to the monitoring sensor device 17.

[0059] Preferably, the monitoring sensor device 17 and / or the light devices 28, 29 are synchronized with the rotation of at least one of the centrifuge chambers 9-12. By "synchronization" we mean the combination of bidirectional timed events to operate the system in unison.

[0060] According to a particularly preferred embodiment, the stroboscope LED or array of LEDs and / or the camera unit are synchronized with the rotation of at least one centrifuge chamber 9-12. In particular, the synchronized operation of the monitoring sensor device 17 ensures that any optical information not related to the centrifuge chamber 9-12 itself is faded out.

[0061] In the preferred embodiment according to Figure 2, the particle loading level 20 represents the range between a particle-free state of the centrifuge chambers 9-12 and a state of maximum particle accumulation, which is the boundary line of particle breakthrough and is defined by the state where particle breakthrough continues as the loading cycle proceeds.

[0062] As also shown in FIG. 2, here preferably the fluidized bed centrifuge 3 comprises a monitoring opening 30 in the monitoring panel, through which the centrifuge chamber contents 23-26 can be monitored. The monitoring opening is assigned and aligned to the optical sensor 18. The fluidized bed centrifuge 3 preferably comprises exactly one monitoring opening 30 in the monitoring panel for monitoring the centrifuge chamber contents 23-26 of all centrifuge chambers 9-12. In this case the monitoring panel with the monitoring opening 30 is preferably fixed while the centrifuge chambers 9-12 rotate about the centrifuge axis 13. Alternatively, according to a particularly preferred embodiment the fluidized bed centrifuge 3 comprises one monitoring opening 30 in the monitoring panel for each centrifuge chamber 9-12, each monitoring opening 30 being aligned to the assigned centrifuge chamber 9-12. In this preferred embodiment each monitoring opening 30 of each monitoring panel moves with the respective centrifuge chamber 9-12 about the centrifuge axis 13.

[0063] For completeness, it can be mentioned that according to Fig. 1 the bioprocessing installation 2 comprises a culture setup 31, in particular a bioreactor, with at least one upstream unit 32 for producing a biological product. As also shown in Fig. 1, the bioprocessing installation 2 comprises a downstream setup 33 with at least one downstream unit 34. This downstream setup 33 is preferably at least one of the group of a filtration setup, a virus inactivation setup, a chromatography setup and / or a virus filtration setup. The at least one downstream unit 34 is preferably at least one of the group of a microfiltration unit, an ultrafiltration unit, a capture chromatography unit, a virus inactivation unit, a diafiltration unit, an intermediate (purification) chromatography unit, a polishing chromatography unit, a virus filtration unit and / or a sterile filtration unit.

[0064] During the proposed tuning routine, at least one parameter of the clarification setup 1 that is tuned is the pump performance, preferably the volumetric flow rate, of at least some of the pumps 5-8 of the pumping device 4, each pump being assigned to one of the centrifuge chambers 9-12.

[0065] The regulation strategy can be realized in numerous ways without departing from the scope of the invention. The regulation strategy is generally intended to control the loading of at least some of the centrifuge chambers 9-12. In the following, in order to briefly explain the invention, a preferred variant is described which controls all of the centrifuge chambers 9-12 according to the regulation strategy. However, all those descriptions given are equally applicable to the variants which control only some of the centrifuge chambers 9-12 according to the regulation strategy.

[0066] Depending on the regulation strategy the particle filling level of the centrifuge chambers 9-12 is controlled. In the present invention this is done on the basis of the particle filling level derived from the monitored sensor data 19, preferably from the image related data 21.

[0067] The adjustment strategy may be aimed at controlling the particle filling level of at least some of the centrifuge chambers 9-12 during the loading cycle so as to reach a target, preferably a target particle filling level or a target time course of the filling level.

[0068] In a particularly preferred embodiment, this means that a target in the form of a maximum filling level is defined which each centrifuge chamber 9-12 is controlled to reach according to a regulation strategy. If the target is reached by one of the chambers 9-12, loading is terminated only for this particular centrifuge chamber 9-12, while the other centrifuge chambers 9-12 continue to be loaded and controlled on the basis of the monitoring sensor data 19. The regulation strategy can also be suitably modified such that all centrifuge chambers 9-12 are filled with particles in a synchronized manner, such that the respective targets are reached by all centrifuge chambers 9-12 at the same time.

[0069] The above mentioned targets may be individual targets for each centrifuge chamber 9-12 in which the particle filling level is controlled. Alternatively, the targets may be shared targets for all centrifuge chambers 9-12 in which the particle filling level is controlled. Here, the invention realizes its full potential in terms of flexibility.

[0070] As indicated above, the regulation strategy involves the particle filling levels of at least some of the centrifuge chambers 9-12 being controlled relative to one another based on monitored sensor data 19, preferably image related data 21. This may include any synchronized control of particle filling levels which may be continuous or discontinuous.

[0071] For example, according to a preferred adjustment strategy, the particle filling levels of the centrifuge chambers 9-12 are controlled to converge based on the monitoring sensor data 19, preferably image related data 21. Additionally or alternatively, according to the adjustment strategy, during the loading cycle, the particle filling levels of the centrifuge chambers 9-12 are controlled to deviate from each other or from the average particle filling level by less than a predefined threshold value T. This is illustrated in FIG. 3b, where the particle filling levels L are displayed for the centrifuge chambers 9-12 during the loading cycle. In this graph, a different line pattern is assigned to each centrifuge chamber 9-12.

[0072] As a comparison, Fig. 3a displays the particle filling levels L of centrifuge chambers 9-12 during the loading cycle without applying the proposed adjustment strategy. It becomes evident that without the proposed adjustment strategy, the filling levels L of centrifuge chambers 9-12 diverge and a rather random distribution of particle filling levels is observed at the end of the loading cycle, compromising not only the efficiency but also the reproducibility of the centrifugation results.

[0073] The proposed solution may also be advantageous from the point of view of balancing the centrifuge 3, and in particular the rotor 15 supporting the centrifuge chambers 9-12, since an uneven filling of particles in the centrifuge chambers 9-12 generally results in an unbalance with respect to the centrifuge axis 13. As mentioned above, in an unbalanced condition the inertial axis 35 of the rotor 15 with respect to the centrifuge axis 13 is offset with respect to the centrifuge axis 13. This unbalanced condition causes vibrations and in extreme cases even mechanical wear or breakage. In any case, the unbalanced conditions impair the efficiency of the centrifuge, since those vibrations affect the fluid flow and reduce the controllability of the process. Advantageously, a balanced operation can be achieved or at least supported by the proposed solution.

[0074] In detail, according to a preferred regulation strategy the volumetric flow rates of the pumps 5-8 assigned to the centrifuge chambers 9-12 are controlled in order to mechanically balance the fluidized bed centrifuge 3 based on a balancing model and on monitored sensor data 19, preferably image related data 21. The balancing model preferably describes the interrelationship between the difference between the particle filling levels of the centrifuge chambers 9-12 and the imbalance of the fluidized bed centrifuge 3.

[0075] According to a particularly preferred embodiment, the balancing model can be adapted, preferably by the electronic process controller 14 and / or a user, to different bioprocesses containing different characteristics, preferably employing different cell broth compositions consisting of different media and / or cell types.

[0076] Different media necessarily contain at least a carbon source, a nitrogen source, water, salts and micronutrients, but various media exist and they may differ with respect to liquid properties such as liquid density, viscosity, etc. The preferred cell types used for bioprocessing are bacteria, plant or mammalian cells, including stem cells or immune cells such as CAR-T cells for cancer therapy, as mentioned above. The selected cells used for bioprocessing are grown in specially designed media, which provide the nutrients required by the respective organism or cell. However, such different cell types also come with differences in cell density, differences in cell viability, differences in cell morphology, differences in cell diameter, etc., which are taken into account by the balancing model.

[0077] Preferably, and only by way of example, the balancing model is adaptable to mechanically balance the fluidized bed centrifuge 3 in bioprocesses employing cell types with higher densities. Since denser particles have a higher mass per volume compared to less dense particles, a larger mass per volume is loaded into the centrifuge chambers 9-12 compared to bioprocesses employing cell types with lower densities. Thus, for the same cell concentration and volumetric flow rate, the centrifuge chambers 9-12 will exhibit a higher mass at a given particle loading level when loaded with a cell type with a higher density. In this case, the volumetric flow rate of at least one pump 5-8 assigned to the respective centrifuge chamber 9-12 is adapted according to a regulation strategy, and preferably the pumps assigned to at least opposing centrifuge chambers 9-12 are adapted relative to each other in order to mechanically balance the fluidized bed centrifuge 3 based on the adapted balancing model. The adaptability of this balancing model allows the supply of cell broths, in particular different cell broths, to the different centrifuge chambers 9-12, which makes the invention particularly flexible.

[0078] In another embodiment, the clarification setup 1 comprises a vibration sensor for detecting the presence and / or amplitude of vibrations in the fluidized bed centrifuge 3, and according to an adjustment strategy, the volumetric flow rates of at least some of the pumps 5-8 assigned to the centrifuge chambers 9-12 are controlled to minimize the amplitude of the vibrations in the fluidized bed centrifuge 3 detected by the vibration sensor. This adjustment strategy can also be applied appropriately when no balancing model is available. However, it can also be applied in addition to the control based on the balancing model. In any case, the reduction of vibrations is preferably achieved with an iterative approach, successively modifying the pump performance, preferably the volumetric flow rates, of the pumps 5-8 and adapting the pump performance accordingly. This makes it even possible to counteract imbalances in the rotor itself or its mechanical support by simply filling the centrifuge chambers 9-12 with particles accordingly.

Claims

1. A method for operating a clarification setup (1) of a bioprocessing installation (2), said clarification setup (1) comprising a fluidized bed centrifuge (3) for the clarification of a cell broth by centrifugation and a pumping device (4) with at least two pumps (5-8) assigned to said fluidized bed centrifuge (3), said fluidized bed centrifuge (3) comprising at least two centrifuge chambers (9-12) rotated about a single geometrical centrifuge axis (13), said bioprocessing installation (2) comprising an electronic process control for controlling at least said fluidized bed centrifuge (3) and said pumping device (4). a fluidized bed centrifuge (3) operated in a forward operation for a particle loading cycle and / or a particle washing cycle and in a reverse operation for a particle discharge cycle, during which the cell broth loaded in the centrifuge chamber (9-12) progresses to form a growing particle accumulation in the centrifuge chamber (9-12), and the clarification setup (1) comprises a monitoring sensor device (17) with at least one, preferably optical, sensor (18) for generating monitoring sensor data which are transmitted to the electronic process control device (14), characterised in that during a monitoring routine, for at least some of the centrifuge chambers (9-12) monitoring sensor data (19) are generated by the monitoring sensor device (17) individually, and during the loading cycle, during an adjustment routine, at least one parameter of the clarification setup (1) is adjusted by the electronic process control device (14) based on the monitoring sensor data (19) so as to control the loading of the cell broth individually for at least some of the centrifuge chambers (9-12) according to a predefined adjustment strategy. method.

2. In the monitoring routine, image-related data (21) representative of an optical image (22) of the centrifuge chamber contents (23-26) is generated as the monitoring sensor data (19) by the monitoring sensor device (17). The method of claim 1.

3. the monitoring sensor data (19) being representative of a particle filling level (20) in each of the centrifuge chambers (9-12) and / or the particle filling level (20) being calculated by the electronic process control device (14) from the monitoring sensor data (19) on the basis of a computational model, 3. The method according to claim 1 or 2.

4. characterised in that the at least one adjusted parameter of the clarification setup (1) is the pumping performance, preferably the volumetric flow rate, of at least some of the pumps (5-8) of the pumping device (4), each assigned to one of the centrifuge chambers (9-12), 4. The method according to claim 1 ,

5. characterised in that during said loading cycle the particle loading level of at least some of said centrifuge chambers (9-12) is controlled according to said regulation strategy, 5. The method according to any one of claims 1 to 4.

6. characterised in that, according to said regulation strategy, during said loading cycle, the particle filling level of at least some of said centrifuge chambers (9-12) is controlled so as to reach a target, preferably a target particle filling level or a target time course of a filling level, 6. The method according to any one of claims 1 to 5.

7. characterised in that the target is an individual target for each centrifuge chamber (9-12) in which the particle filling level is controlled or the target is a shared target for all centrifuge chambers (9-12) in which the particle filling level is controlled, 7. The method according to any one of claims 1 to 6.

8. characterised in that according to said regulation strategy the particle loading levels of at least some of said centrifuge chambers (9-12) are controlled relative to one another, 8. The method according to any one of claims 1 to 7.

9. According to said adjustment strategy, the particle filling level of at least a part of said centrifuge chambers (9-12) is controlled to converge based on said monitoring sensor data (19), preferably image-related data (21).

9. The method according to any one of claims 1 to 8.

10. characterised in that according to said adjustment strategy the particle filling levels of at least two centrifuge chambers (9-12) are controlled based on said monitoring sensor data (19), preferably image-related data (21), to deviate from each other or from an average particle filling level by less than a predefined threshold value, 10. The method according to any one of claims 1 to 9.

11. According to said adjustment strategy, the volumetric flow rates of at least two pumps (5-8) assigned to a centrifuge chamber (9-12) are controlled on the basis of a balancing model based on said monitoring sensor data (19), preferably image-related data (21), in order to mechanically balance said fluidized bed centrifuge (3), preferably said balancing model describing the interrelationship between the difference between the particle filling levels of at least two centrifuge chambers (9-12) and an imbalance of said fluidized bed centrifuge (3).

11. The method according to any one of claims 1 to 10.

12. characterised in that the clarification setup (1) comprises a vibration sensor for detecting the presence and / or amplitude of vibrations in the fluidized bed centrifuge (3), and according to the regulation strategy the volumetric flow rates of at least some of the pumps (5-8) assigned to the centrifuge chambers (9-12) are controlled so as to minimise the amplitude of vibrations in the fluidized bed centrifuge (3) detected by the vibration sensor, 12. The method according to any one of claims 1 to 11.

13. A clarification setup (1) of a bioprocessing installation (2) for carrying out the method according to any one of claims 1 to 12, said clarification setup (1) comprising a fluidized bed centrifuge (3) for the clarification of a cell broth by centrifugation and a pumping device (4) with at least one pump (5-8) assigned to said fluidized bed centrifuge (3), said fluidized bed centrifuge (3) comprising at least two centrifuge chambers (9-12), each of which is rotated around a single geometrical centrifuge axis (13), said bioprocessing installation (2) comprising a controller (4) for controlling at least said fluidized bed centrifuge (3) and said pumping device (4). said fluidized bed centrifuge (3) being operated in a forward operation for a particle loading cycle and / or a particle washing cycle and in a reverse operation for a particle discharge cycle, during said particle loading cycle a cell broth loaded in said centrifuge chamber (9-12) progresses to form a growing particle accumulation in said centrifuge chamber (9-12), said clarification setup (1) comprising a monitoring sensor device (17) with at least one, preferably optical, sensor (18) for generating monitoring sensor data which are transmitted to said electronic process control device (14), characterised in that during a monitoring routine, for at least some of the centrifuge chambers (9-12) monitoring sensor data (19) are generated by the monitoring sensor device (17) individually, and during the loading cycle, during an adjustment routine, at least one parameter of the clarification setup (1) is adjusted by the electronic process control device (14) based on the monitoring sensor data (19) so as to control the loading of the cell broth individually for at least some of the centrifuge chambers (9-12) according to a predefined adjustment strategy. Clarification Setup (1).

14. 14. The electronic process control device of claim 13, further comprising: Characterized in that it is designed to carry out the method according to any one of claims 1 to 12, Electronic process control equipment.

Images