Methods for performing bioprocessing on liquid immune or naive cell cultures to obtain processed cell cultures

Abstract

The present invention relates to a method for performing a bioprocess on a liquid immune or naive cell culture to obtain a processed cell culture, wherein the processed cell culture is for autologous or allogeneic cell therapy, the bioprocess being performed in an integrated bioprocessing system (1), the bioprocess comprising a series of processing steps, each of the processing steps comprising at least one operation, the bioprocess system (1) comprising a base structure (3) and preconfigurable cartridges (4), the bioprocess system (1) performing the operations of the bioprocess by interaction between the base structure (3) and the cartridges (4), the cartridges (4) and the base structure (3) comprising matching standardized interfaces (6) for interaction between the base structure (3) and each cartridge (4), and the bioprocess system (1) performing at least two operations of the bioprocess inside at least two differently preconfigured cartridges (4) by interaction between the base structure (3) and the cartridges (4) via the same base structure interface (7) and / or a plurality of identical base structure interfaces (7) and matching cartridge interfaces (8).

Description

[Technical Field]

[0001] The present invention relates to a method for performing a bioprocess on a liquid immune or naive cell culture and obtaining a processed cell culture according to claim 1, a cartridge according to claim 35, a base structure according to claim 37 and a bioprocessing system according to claim 38.

[0002] The term "bioprocessing" currently refers to biotechnological processes, particularly those involving the use of naive cell cultures, including immune cell cultures or stem cell cultures. One or more processing steps may be performed on each cell culture. Thus, bioprocessing in this sense can refer to a manufacturing process that includes a series of processing steps performed on cell cultures that ultimately result in a final product.

[0003] The proposed method can be used in the field of cell and gene therapy, including the allogeneic or autologous production of genetically modified immune cells. For example, this method can be applied to the production of autologous T cells modified to express chimeric antigen receptors (CARs). These cells can be used to treat various types of malignant blood diseases, including different types of leukemia (blood cancer). Other cell therapies based on naive cells, particularly stem cells and their derivatives, are also attracting interest.

[0004] Process flexibility is particularly important in biotechnology processes involving the use of liquid immune or naive cell cultures. The starting material can be highly heterogeneous in terms of its composition, for example, because each patient's condition (e.g., regarding disease progression or the genetic makeup and history of their immune system) varies. The starting material can also be heterogeneous in terms of its composition because cell cultures from different donors are combined into a single initial starting material. Therefore, depending on the source of the liquid cell culture, various parameters can vary, including, for example, the type and concentration of different cell types, overall cell viability and vitality, and / or the amount and type of impurities within the liquid cell culture. Furthermore, depending on the bioprocess being performed, the cell culture may contain different amounts of different types of immune cells (e.g., T cells, dendritic cells, or immune cells at different developmental stages, including naive T cells). For reasons discussed above, the order of processing steps performed on each liquid immune cell culture must be tailored to the individual characteristics of the cell culture. Additionally, certain steps of the bioprocess may need to be flexibly adapted and tailored to each cell culture. For example, the types of genetic modifications of cell cultures may differ because patients may respond differently to certain genetic modifications. Therefore, bioprocesses differ in the methods of carrying out the genetic modifications. Also, for example, a bioprocess involving genetic modification of immune cell cultures requires a different order of processing steps than that required for a bioprocess without genetic modification of immune cell cultures. Not only can steps be flexibly adapted, but steps can also be omitted entirely. Therefore, the method of carrying out a bioprocess on liquid immune cell cultures must be flexibly adaptable.

[0005] While process flexibility is crucial, cost-efficiency is also a key aspect in the processing of liquid immune or naive cell cultures. This is due to stringent regulatory requirements. For example, process operators must be highly skilled, especially when performing manual processes. Additionally, sterility must be maintained during the manufacturing process because processed cells must be viable and free of contaminants when administered to recipients. For one thing, for the reasons mentioned above, the overall cost of each bioprocess performed is very high. Furthermore, approaches that use a single device to sequentially perform all processing steps on the initial cell culture typically result in a large facility footprint, since one device is required to process one cell culture at a time. Furthermore, throughput is low because only one liquid cell culture is processed within a single device. Furthermore, because the order of processing steps is typically pre-specified, for example, by the installed tubing set, adapting the device to change the order or type of processing steps performed is quite difficult. Nevertheless, these end-to-end systems provide a system in which all media is maintained within a closed internal volume, which supports sterility maintenance.

[0006] Cost-effectiveness is particularly relevant to operational costs. Therefore, it is desirable to be able to process two or more cell cultures in parallel. This also increases throughput, which is important considering that processing a single initial cell culture can take days or even weeks. However, when processing two or more cell cultures in parallel, it is necessary to ensure that cross-contamination of different cell cultures does not occur.

[0007] Another important aspect is redundancy, which also relates to the robustness and reliability of the process.As mentioned above, especially when applied to cancer treatment, the processing of cell culture may take a considerable amount of time, and patients usually need urgent treatment.Therefore, it is necessary to ensure that the process does not fail.If the process fails, it may have to be repeated, which requires additional time and / or requires additional starting material.However, especially in autologous approach, additional starting material may not be obtained from patients.

[0008] The known prior art on which the present invention is based is WO 2021 / 212124, which relates to a method according to the general part of claim 1. This publication discloses a method for the modular parallel processing of liquid immune cell cultures in an integrated bioprocessing system. The method comprises a series of processing steps carried out within specific unit operation stations of the bioprocessing system. While the bioprocessing system exhibits a degree of standardization in the form of racks that receive all components necessary for the performance of unit operations, the unit operation stations themselves are each highly customized with respect to the unit operations to be performed.

[0009] Therefore, the complete configuration of each unit operation station varies depending on the type of unit operation performed at each unit operation station. As a result, the infrastructure provided at each unit operation station (e.g., reservoirs for raw materials and / or waste) also varies depending on the unit operation performed at each unit operation station. Furthermore, the interaction between the transport system and each unit operation station varies depending on the type of unit operation performed at each unit operation station. As a result, while the high degree of customization of unit operation stations allows for great flexibility in defining process steps, the potential for efficiency gains through automation remains relatively low.

[0010] The problem underlying the present invention is to improve the known method in such a way that it allows for increased automation possibilities without compromising flexibility regarding the definition of the process steps.

[0011] The above object is solved by the features of the characterizing part of claim 1.

[0012] The main realization of the present invention is that by providing a standardized architecture based on individually pre-configured cartridges, a large number of different processing steps can be performed in a flexible manner by an integrated bioprocessing system while maintaining sterility, thereby increasing the efficiency and flexibility of the entire bioprocess. The main idea is to separate the parts of the system that can be reused from the liquids and liquid-containing single-use components, and to provide a modular bioprocessing system in which multi-use and single-use components can be combined as needed.

[0013] A method is proposed for performing a bioprocess on a liquid immune or naive cell culture to obtain a processed cell culture, the processed cell culture being for autologous or allogeneic cell therapy, the bioprocess being performed in an integrated bioprocessing system, the bioprocess comprising a series of processing steps, each of the processing steps comprising at least one operation, the bioprocessing system comprising a base structure and preconfigurable cartridges, the bioprocessing system performing the operations of the bioprocess by interaction of the base structure with the cartridges, the cartridges and the base structure comprising matching standardized interfaces for interaction of the base structure with each cartridge, and the bioprocessing system performing at least two operations of the bioprocess inside at least two differently preconfigured cartridges by interaction of the base structure with the cartridges via the same base structure interface and / or a plurality of identical base structure interfaces and matching cartridge interfaces.

[0014] The term "different" should be understood to refer to functionally significant differences rather than, for example, purely visual deviations. Different pre-configured cartridges may have different numbers of tubes, substantially different tube lengths, different functional devices, different numbers of inlets or outlets, different containers for receiving and / or dispensing liquids, etc.

[0015] The term "standardized" means that differently configured cartridges for different operations can be used through the same base structural interface. Preferably, the cartridge interfaces are identical.

[0016] The term "pre-configured" means that each cartridge is configured outside the bioprocessing system, for example, manually or by the cartridge manufacturer, or that the cartridge is automatically configured by the bioprocessing system with the addition of components. Configuration should not be understood to relate only to digital configuration or minimal modification, or particularly to in-use modification of the cartridge. Pre-configuration mechanically changes the state of the cartridge from an unusable state to a usable state. In particular, pre-configuration includes adding, particularly modifying (removing and adding), cartridge fluidic structures.

[0017] Claims 2 to 4 relate to preferred combinations of bioprocesses, bioprocessing steps and operations carried out by the bioprocessing system.

[0018] Claims 5 and 6 describe preferred embodiments of the cartridge.

[0019] Claims 7 to 9 describe preferred embodiments of the interface.

[0020] Claim 10 describes a receptacle that is preferably used by a bioprocessing system.

[0021] Claims 11 to 28 relate to preferred embodiments of different cartridges adapted for multiple process steps.

[0022] Claims 29 to 34 relate to various details of the method.

[0023] Another equally important teaching according to claim 36 relates to a cartridge for use in the proposed method.

[0024] All explanations given regarding the proposed method are fully applicable.

[0025] Another equally important teaching according to claim 38 relates to a base structure used in the proposed method.

[0026] All explanations given regarding the proposed method are fully applicable.

[0027] Another equally important teaching according to claim 39 relates to a bioprocessing system for use in the proposed method.

[0028] All explanations given regarding the proposed method are fully applicable.

[0029] In the following, embodiments of the invention will be described with reference to the drawings. [Brief explanation of the drawings]

[0030] [Figure 1] FIG. 1 is a schematic diagram of the proposed integrated bioprocessing system. [Figure 2] a) A perspective view of the cartridge itself without the cartridge fluidic structure, and b) an explosive representation of the cartridge with the cartridge fluidic structure and unit operation station. [Figure 3] FIG. 2 shows a unit operation station of the bioprocessing system of FIG. 1 during the performance of a unit operation. [Figure 4]4A-4B are cross-sectional views of the bioprocessing system of FIG. 1 a) along line IV-IV and b) along line VV. [Figure 5] A diagram showing the unit operation stations of the bioprocessing system of Figure 1 during a standard routine in sequences a) to e). [Figure 6] 6A and 6B show the unit operation station of FIG. 5 a) during the connection process and b) during the disconnection process. [Figure 7] A figure showing the unit operation station of Figure 5 in sequence a) to b) during the transport of a cartridge from the drive position to cartridge waste storage. [Figure 8] Schematic diagram of the closing connection process in sequence a) to e). [Figure 9] FIG. 10 is a diagram showing an example of a cartridge. [Figure 10] 1 illustrates a functional interface of a cartridge and a receptacle connectable to the functional interface.

[0031] The integrated bioprocessing system 1 shown in the drawings is preferably adapted to carry out a bioprocess for the production of genetically modified T cells. Here, the T cells are genetically modified to express a chimeric antigen receptor (CAR). Consequently, the term "CAR-T cells" refers to T cells genetically modified to express a CAR. The genetically modified CAR-T cells, which are the product of the bioprocess, can be administered to a patient and used to initiate or restart cancer treatment in the patient. By carrying out the bioprocess, the initial immune cell culture is gradually processed. All descriptions provided are primarily directed to such a bioprocess. However, it may be noted that these descriptions also apply equally well to other bioprocesses.

[0032] The term "liquid immune cell culture" should be understood broadly and refers to an immune cell culture containing at least one type of immune cell suspended as particles in any type of liquid. As explained below, a liquid immune cell culture may contain other cell types that are not immune cells. Thus, the term "liquid immune cell culture" refers to a liquid immune cell culture at any stage during a bioprocess. As a result, the types and fractions of immune cells present in the liquid immune cell culture change during the applied bioprocess as certain immune cells are enriched or depleted from the liquid immune cell culture and / or the immune cells are genetically modified.

[0033] The term "immune cells" generally refers to various types of white blood cells. Thus, the term "immune cells" includes various cells, such as, but not limited to, dendritic cells, T lymphocytes (also known as T cells), B lymphocytes, natural killer cells, macrophages, etc. Immune cells may also include subtypes of immune cells, such as tumor-infiltrating lymphocytes or various types of T cells. Subtypes of certain immune cells can be classified based on the type of antigen present on the cell surface. Thus, the term immune cells may refer, for example, to T cells containing the surface antigen CD4 ("CD4+ T cells"). Typically, certain immune cells, such as T cells, and preferably certain subtypes of immune cells, such as CD4+ T cells, are selectively enriched by bioprocesses, while other immune cells, such as macrophages, and / or other cell types that are not immune cells, such as erythrocytes, and / or other subtypes of immune cells, such as CD8+ T cells, are depleted from the liquid immune cell culture. The immune cells that are enriched are referred to as target immune cells, and all other components that are depleted from the liquid immune cell culture are referred to as “impurities.” Additionally, as described above, the target immune cells can be genetically modified.

[0034] The term "naive cells" refers to cells that can still differentiate into different target cell types. In particular, stem cells and their derivatives before they have fully differentiated into a specific cell type are naive cells. This term also includes naive immune cells.

[0035] The term "liquid" should also be understood broadly and refers to any liquid and / or particle-containing liquid that is processed within the integrated bioprocessing system 1. Thus, the term liquid may refer to media, including wash buffers and cell growth feed media, waste, liquid immune cell cultures, by-products obtained during a bioprocess, samples, and / or initial immune cell cultures.

[0036] The term "waste" refers to any liquid and / or particle-containing liquid obtained during a bioprocess, and each liquid and / or particle-containing liquid may be discarded and not used further. However, waste may be stored and, in some cases, reused before being discarded. For example, if the number of T cells extracted from a cell culture is too low, more T cells may be extracted from the waste.

[0037] The term "initial immune cell culture" refers to a liquid immune cell culture before the first processing step of a bioprocess is applied. The initial immune cell culture can be derived from a variety of sources. In an approach often referred to as "autologous cell therapy," the initial immune cell culture is obtained from a donor who is also the recipient of the product after the bioprocess is fully implemented. In "allogeneic cell therapy," the initial immune cell culture can be derived from at least one donor who is not the recipient of the product. The initial immune cell culture may be derived from more than one donor and / or used for more than one recipient. In this case, immune cell cultures obtained from different donors are combined into a single initial immune cell culture.

[0038] Preferably, the initial immune cell culture is obtained by a process called "leukapheresis." In leukapheresis, immune cells are obtained from a patient. Additionally or alternatively, the initial immune cell culture may be obtained from the patient's tissue. As noted above, the initial immune cell culture may be obtained from one or more donors who are not the patient. Additionally, in some cases, other cells may be obtained from the same patient, such as tumor cells used in a bioprocess to modulate immune cell behavior.

[0039] Depending on the source of the initial immune cell culture and the bioprocess being performed, the initial immune cell culture, particularly the type, amount, and distribution of impurities, and the target immune cells, may vary. Additionally, the initial immune cell culture may be obtained from a frozen source, or the initial immune cell culture may be frozen and thawed as needed.

[0040] Currently, it is preferred that any initial immune cell culture used in a bioprocess be used for only a single patient or a small number of patients, for example, up to 10 patients. Otherwise, the currently described bioprocess will only produce cell cultures for a single patient or a small number of patients, while cell cultures for other patients will be obtained from a different bioprocess. Therefore, the proposed integrated bioprocessing system 1 is used for small-scale bioprocesses. The bioprocessing system 1 itself may be larger, running multiple bioprocesses in parallel.

[0041] The term "processing step" refers to a distinct step performed as part of a bioprocess using a liquid immune cell culture. The type and order of the processing steps performed vary depending on the bioprocess and the type of immune cell culture being performed on each liquid immune cell culture. Depending on the parameters mentioned, various processing steps can be combined in any given order. Additionally or alternatively, the configuration of processing steps may differ and / or processing steps may be repeated and / or omitted. Each processing step includes at least one operation. An operation is the smallest unit of processing in a bioprocess with a defined start and end. Typically, a processing step includes several operations, such as pumping, mixing, centrifugation, etc. When one or more operations of a processing step are described herein, it is preferred that they include most or all of the associated operations of each processing step.

[0042] A method is proposed for performing a bioprocess on a liquid immune or naive cell culture to obtain a processed cell culture. The processed cell culture can be used for cancer therapy, as described above. The method can also include performing multiple bioprocesses on multiple liquid immune or naive cell cultures to obtain multiple processed cell cultures. Preferably, the processed cell cultures are for autologous or allogeneic cell therapy. Each bioprocess is performed on a different cell culture, and all processing steps performed on one cell culture should be understood as a single bioprocess. Multiple cell cultures can be used to treat patients. Alternatively, multiple cell cultures can be obtained from a single input patient material to evaluate and compare different production processes. The optimal production process identified is then used for subsequent therapeutic manufacturing.

[0043] The bioprocess is carried out in an integrated bioprocessing system 1, for example, as shown in Figure 1. The integrated bioprocessing system 1 of Figure 1 is merely an exemplary embodiment, and other embodiments may vary significantly, as will be apparent from the following description.

[0044] A bioprocess comprises a series of processing steps, each of which comprises at least one operation. In the following, we also refer to unit operations. A unit operation is a collection of operations performed in a unit operation station 2, which is present in some embodiments. A unit operation may comprise a portion of a processing step, or even multiple processing steps.

[0045] The bioprocessing system 1 comprises a base structure 3 and a preconfigurable cartridge 4. The cartridge 4 can be preconfigured with a cartridge fluidic structure 5 for at least one operation. Preconfiguration is preferably performed outside the bioprocessing system 1. On this basis, depending on the configuration of the cartridge fluidic structure 5, almost any operation can be realized with the preconfigured cartridge 4. In this way, each cartridge fluidic structure 5 can be highly individualized for different bioprocesses, and in particular also for different liquid cell cultures.

[0046] FIG. 2a shows the cartridge 4 in an unconfigured state without the cartridge fluid structure 5, and FIG. 2b shows the cartridge 4 pre-configured with the cartridge fluid structure 5 for at least one operation.

[0047] The bioprocessing system 1 performs bioprocess operations through the interaction of the base structure 3 and the cartridges 4. That is, the base structure 3 forms the basis for performing a bioprocess, and the cartridges 4 form the basis for flexibly performing different bioprocesses. The cartridges 4 and the base structure 3 have matching standardized interfaces 6 for the interaction between the base structure 3 and each cartridge 4. Due to the standardization, flexibility is achieved without the need to adapt the base structure 3 for different operations.

[0048] Thus, the bioprocessing system 1 performs at least two operations of a bioprocess inside at least two differently pre-configured cartridges 4 through the interaction of the base structure 3 and cartridges 4 via the same base structure interface 7 and / or multiple identical base structure interfaces 7 and matching cartridge interfaces 8. It is possible, but not necessary, to use the same base structure interface 7 for the two operations. The cartridge interfaces 8 may be different as long as they are compatible with the base structure interface 7. For example, the cartridge 4 may simply have holes where mechanical energy transmission elements can enter if the cartridge 4 does not require any mechanical energy in the pre-configuration.

[0049] The term "interface" includes all functional, at least partially mechanical, connections between the base structure 3 and the cartridge 4. There is at least a base structure interface 7 that supports at least two operations. The base structure 3 may have multiple base structure interfaces 7 for performing operations inside the cartridge 4. These base structure interfaces 7 directly affect the actual operations. The base structure interfaces 7 may include electrical connections that supply electrical energy to the cartridge 4, and / or cable-coupled electrical connections that supply signals to the cartridge 4, and / or mechanical connections that supply mechanical energy to the cartridge 4, and / or pneumatic connections that supply pneumatic energy to the cartridge 4.

[0050] The base structure 3 may comprise a number of different base structure interfaces 7 for performing operations of different subsets of the processing steps. Preferably, these base structure interfaces 7 are at least partly identical to one another, for example comprising the same electrical and / or mechanical and / or pneumatic and / or signal connectors. However, they may comprise further distinctive elements, such that not all operations can be performed by all base structure interfaces 7. In particular, functional elements may be present as part of the base structure interfaces 7 only in some locations, allowing the base structure interfaces to be adapted to the functional interfaces.

[0051] There may also be different additional base structure interfaces for auxiliary functions, in particular a standardized transport and placement interface 35. The transport and placement interface 35 here preferably comprises one or more transport elements for transporting cartridges 4 and / or receptacles 15 and / or containers 30 between the base structure interfaces 7.

[0052] Bioprocessing steps other than those described may be part of the bioprocess and may be performed by the bioprocessing system 1.

[0053] Here, preferably, the bioprocess is carried out by the bioprocessing system 1 on at least three, preferably at least four, differently individualized cartridges 4. Preferably, no more than eight differently individualized cartridges 4 are used in a single bioprocess.

[0054] The plural term "cell cultures" also includes what may be referred to in the singular as a "cell culture."

[0055] The embodiments shown in the drawings will now be described, followed by a discussion of further variations, not all of which are shown in the drawings. Figure 1 shows an integrated bioprocessing system 1, and Figure 2 shows a unit operation station 2, which is one way of flexibly providing the main parts of the bioprocessing system 1. Figure 3 shows the performance of a unit operation on the unit operation station 2. Figure 4 shows how the bioprocessing system 1 can be constructed from multiple such unit operation stations 2. The unit operation station 2, together with other components such as a transport mechanism, builds up a base structure 3 and interacts with a cartridge 4.

[0056] In the embodiment shown in the drawings, a standard routine is defined to perform one or more operations. According to the standard routine, one of the cartridges 4 is transported from the cartridge storage unit 9 of the unit operation station 2 to the drive position 10 of the cartridge drive unit 11 of the unit operation station 2 by the local transport mechanism 12 of the unit operation station 2, which is only shown in FIG. 2b (sequence diagrams 5a, 5b, 5c, and 5d). This local transport mechanism 12 may, for example, comprise a linear actuator and / or several linear actuators, and / or a combination of a linear actuator with a multi-axis robotic manipulator, a conveyor belt, etc. The cartridge 4 is then operably coupled to the cartridge drive unit 11, which is a standardized interface 6 between the base structure 3 comprising the unit operation station 2 and the cartridge 4.

[0057] Preferably, the performance of any one of the unit operations includes the performance of the standard routine described above, which provides for the transfer of each cartridge 4 from the cartridge storage unit 9 to the cartridge drive unit 11 and the operative coupling of the cartridge 4 with the cartridge drive unit 11. The performance of subsequent unit operations is then individualized according to the pre-configuration of the cartridge 4. Neither the unit operation station 2 nor the unit operations are necessarily present in all embodiments.

[0058] The operable coupling between the cartridge 4 and the cartridge drive unit 11 allows the cartridge 4 to be a completely passive component without any kind of actuator. This operable coupling allows any actuation to be transmitted from the cartridge drive unit 11 to the cartridge 4. However, it is generally possible for the cartridge 4 to include additional actuators.

[0059] The term "operably coupled" should be understood broadly. Preferably, this means a functional engagement between the cartridge drive unit 11 and the cartridge 4. Thus, the term "coupled" may refer to a physical engagement between both components. Additionally or alternatively, it may refer to an indirect connection established between the two components (e.g., by magnetic actuation). The cartridge drive unit 11 may actuate the functional device 13 of the cartridge 4. This is all performed via the base structural interface 7 and the cartridge interface 8.

[0060] According to a preferred embodiment, it is proposed that at least two, and preferably all, of the cartridges 4 are identical to one another with the exception of their respective cartridge fluidic structures 5, which may be configured for each unit operation. That is, the cartridges 4 may (but need not) be offset from one another as far as their respective cartridge fluidic structures 5 are concerned. Using identical cartridges 4 in this sense allows standardization of not only the transport of the cartridges 4, but also the transfer of liquids to and from the cartridges 4.

[0061] According to another embodiment, it is proposed that at least one unit operation station 2 comprises at least one transfer position 14 shown in Figure 3. Here, preferably, the transfer position 14 is provided by a surface or pad for receiving a receptacle 15. Thus, a pre-configured receptacle 15 with a receptacle fluidic structure 16 for containing a liquid, preferably an immune or naive cell culture, is transported to the transfer position 14 by the overall transport mechanism 17 shown in Figure 1 for transferring the liquid between the cartridge 4 transported to the drive position 10 and the receptacle 15 and / or for performing the respective unit operation on the liquid contained in the receptacle 15.

[0062] Preferably, each unit operation station 2 herein comprises four transfer positions 14. As shown in Figure 1, and in the preferred embodiment to that extent, each transfer position 14 is rectangular in shape, with two transfer positions 14 arranged adjacent to each other. Preferably, two pairs of transfer positions 14 are arranged opposite each other, with the drive position 10 located between each pair of transfer positions 14.

[0063] As will be explained below, the receptacle 15 located at the transfer position 14 must be aligned with the cartridge 4 located at the drive position 10. For alignment, an entire transport mechanism 17, preferably a robotic mechanism 18, may be used, or the transfer position 14 may include a position adjustment mechanism for aligning the receptacle 15 with the cartridge 4.

[0064] According to one embodiment, it is proposed that at least two, and preferably all, of the receptacles 15 are identical to one another except for their respective receptacle fluid structures 16, which may be configured for the respective liquids contained therein. That is, as far as the respective receptacle fluid structures 16 are concerned, the receptacles 15 may (but need not) be offset from one another. The liquids contained in the receptacle fluid structures 16 may differ depending on the process steps performed. That is, the complete handling of the receptacles 15, including transport, establishing fluid connections, etc., may be standardized for all receptacles 15. An example of these identical receptacles 15 is shown in FIG. 3.

[0065] According to one embodiment, the integrated bioprocessing system 1 comprises a number of unit operation stations 2, and it is preferably proposed that the transfer positions 14 of the unit operation stations 2 are arranged in a first plane 19, and that the receptacles 15 are transported to and from the transfer positions 14 in this first plane 19 by an overall transport mechanism 17. The first plane 19 here is preferably aligned horizontally. By arranging the transfer positions 14 in the first plane 19, the transport of the receptacles 15 to and from the transfer positions 14 is simplified. In addition, as will be explained below, the receptacles 15 are arranged in the same plane as the cartridges 4 that are subsequently arranged in the drive position 10.

[0066] Preferably, at least some of the multiple unit operation stations 2 are arranged in a unit operation station slice 20. Each unit operation station slice 20 preferably has a separate transport device, such as wheels, for moving the unit operation station slice 20 in and out of the integrated bioprocessing system 1. The ability to remove a unit operation station slice 20, including multiple unit operation stations 2 arranged therein, from the integrated bioprocessing system 1 particularly simplifies maintenance of the unit operation stations 2, as described further below. Furthermore, a unit operation station slice 20 can be removed from or installed in the integrated bioprocessing system 1 without affecting the operability of the remaining unit operation stations 2 in the integrated bioprocessing system 1. This is particularly advantageous when access to a portion of the integrated bioprocessing system 1 is required for maintenance.

[0067] According to one embodiment, it is proposed that at least one bioprocess is a closed bioprocess, in which all liquids involved in each bioprocess are kept in a closed internal volume. In the illustrated embodiment, this closed structure is preferably achieved by performing at least part of the liquid handling involved in each bioprocess in tubes.

[0068] To facilitate the realization of a closed bioprocess, it is proposed that for the transfer of liquid between the cartridge 4 and the receptacle 15, the cartridge transfer tube 21 of the cartridge 4 and the receptacle transfer tube 22 of the receptacle 15 are connected in a closed connection process by a tube connection system 23. This is shown in Figure 6a. Preferably, the connection process is performed by a tube connection system, in particular a tube welding system 24.

[0069] The connection process is shown in Figure 8. The connection process is performed by the tube connection system 23, in particular by a tube handling device (not shown) of the tube connection system 23. It also preferably includes a step of positioning the tubes to be connected relative to the rest of the tube connection system 23 (sequence Figures 8a-8b)) and a subsequent step of welding the tubes to be connected by the tube connection system 23 (Figure 8d)). After welding, the tubes are connected at the welding position 57, as shown in Figure 8e). Further preferably, the connection process includes a step of trimming the tubes to be connected (Figure 8b)) before the welding step. Preferably, the trimming of the tubes is performed by a blade 58, as shown in Figure 8c).

[0070] As shown in FIG. 3 , the cartridge transfer tubes 21 and the receptacle transfer tubes 22 are provided by the cartridge fluid structure 5 and the receptacle fluid structure 16. Preferably, the cartridge fluid structure 5 and the receptacle fluid structure 16 are pre-configured so that each cartridge transfer tube 21 is positioned within a predetermined cartridge anchor point 25 provided by the cartridge frame 26, and each receptacle transfer tube 22 is positioned within a predetermined receptacle anchor point 27 provided by the receptacle 15. This is shown, for example, in FIG. 3 . Preferably, the cartridge anchor points 25 and the receptacle anchor points 27 are positioned so that one cartridge anchor point 25 faces one receptacle anchor point 27 when each cartridge 4 is in the drive position 10 and each receptacle 15 is in the transfer position 14.

[0071] 3, the cartridge transfer tube 21 preferably projects from the cartridge anchor point 25 toward the transfer location 14 where the connected receptacle 15 is located. It is also preferred that the receptacle transfer tube 22 projects from the receptacle anchor point 27 toward the cartridge anchor point 25 where the cartridge transfer tube 21 is located. The welding system 24 connects the receptacle transfer tube 22 and the cartridge transfer tube 21 in a closed connection process.

[0072] In addition, as shown in FIG. 6b, the welding system 24 is preferably designed to also perform a cutting process. Here, the cut transfer tube is preferably closed and cut at each end so that the internal volume of the tube remains sterile. It may also be noted that using such a welding system 24, multiple welds can be made sequentially on the same transfer tube. This provides flexibility, for example, for connecting the same receptacle transfer tube 22 to different cartridge transfer tubes 21 sequentially.

[0073] According to one embodiment, it is proposed that at least two of the above bioprocesses are carried out at least partially simultaneously by an integrated bioprocessing system 1, preferably coordinated by an electronic process control 28. This is shown in Figure 4a, where two cartridges 4 of two separate unit operation stations 2 are transported to their respective drive positions 10.

[0074] In this illustrated, and to that extent preferred, embodiment, the cartridge 4 comprises a cartridge carrier 29 that receives the components of the cartridge fluidic structure 5. An advantage thereof is that the cartridge carrier 29 provides a standardized structure for receiving the cartridge fluidic structure 5. The size of the cartridge carrier 29 is therefore preferably the same for all unit operation stations 2 and is therefore independent of the particular unit operation being performed at the unit operation stations 2. The cartridge fluidic structure 5 can therefore be positioned as desired within the cartridge carrier 29, which provides a standardized interface 6 to the cartridge frame 26 and the cartridge drive unit 11. Furthermore, the cartridge carrier 29 does not come into contact with any liquid handled at the unit operation stations 2. Therefore, the cartridge carrier 29 can be reused after the cartridge 4 has been used in a unit operation. Furthermore, the use of the cartridge carrier 29 to receive the components of the cartridge fluidic structure 5 ensures that any liquid that may leak from the cartridge fluidic structure 5, for example if the cartridge fluid reservoir 30 is broken, is contained within the cartridge 4 and does not contaminate other elements, such as the cartridge drive unit 11 of the integrated bioprocessing system 1.

[0075] As shown in Figure 2b, the cartridge carrier 29 may include a drive recess 31 that allows the drive structure of the cartridge drive unit 11 to engage with components of the cartridge fluidic structure 5. For example, as shown in Figure 2a, the cartridge carrier 29 may include a drive recess 31 at a position where a pump head of a peristaltic pump 32 is located. In order to actuate the components of the cartridge fluidic structure 5 via the interfaces provided by the base structure 3, it is important that the drive recess 31 in the cartridge carrier 29 is aligned with the respective interfaces of the base structure 3.

[0076] Preferably, the material of the cartridge carrier 29 is a multi-use material such as plastic or the like. Furthermore, as mentioned above, each cartridge carrier 29 is preferably standardized in terms of size. It is also preferred that the longitudinal and lateral dimensions of the cartridge carrier 29 correspond to the cartridge drive unit 11. As will be described below, the cartridge drive structure 33 of the cartridge drive unit 11 provides an interface that engages with components of the cartridge fluidic structure 5.

[0077] As will be apparent, the base structure 3 can be comprised of multiple, even interchangeable, components, where, preferably, during use of the bioprocessing system 1, the base structure 3 is an interconnected structure. The base structure 3 is not comprised of unrelated, unconnected components.

[0078] All liquids involved in the bioprocess are preferably kept within one or more closed internal volumes. This ensures that the sterility of all liquids is maintained. Furthermore, it ensures that cross-contamination between different cell cultures does not occur. This is particularly important when processing multiple cell cultures in parallel. Furthermore, the environmental requirements for sterility are significantly reduced. The term "closed internal volume" currently means that all described liquids are maintained and conducted within a volume isolated from the atmosphere. The term "atmosphere" currently refers to the volume outside the closed internal volume. This ensures that sterility within the closed internal volume is maintained. As explained above, maintaining sterility is an important aspect of the manufacturing process. A closed internal volume can be easily achieved if at least part of the handling of the liquids involved in each unit operation is performed within tubing. Preferably, the integrated bioprocessing system 1 includes an enclosure 34 in which the bioprocess is carried out. The interior of the enclosure 34 may be provided with a sterile atmosphere, but is preferably not as sterile as the closed internal volume.

[0079] According to one embodiment, it is proposed that the two operations of the bioprocess are operations of different processing steps and / or that the bioprocessing system 1 performs at least two, preferably at least three, more preferably at least four operations that are part of different bioprocesses in parallel within the same preconfigured cartridge 4.

[0080] Additionally or alternatively, the bioprocessing system 1 can perform at least two, preferably at least three, more preferably at least four operations in parallel in different pre-configured cartridges 4 that are part of different bioprocesses, in particular different processing steps of different bioprocesses.

[0081] According to one embodiment, it is proposed that the bioprocessing system 1 carries out at least two bioprocesses, preferably at least three, more preferably at least five, more preferably at least ten bioprocesses, in particular in parallel, each comprising at least two, preferably at least three, more preferably at least four different processing steps, and that the bioprocessing system 1 carries out at least one operation for each processing step, and that the bioprocessing system 1 carries out these operations in at least two, preferably at least three, more preferably at least four differently preconfigured cartridges 4 for each bioprocess. Preferably, all these operations are carried out automatically, without manual modification of the base structure 3.

[0082] According to one embodiment, it is proposed that the manipulation comprises one or more of the process steps "enrichment" and / or "selection" and / or "activation" and / or "loading" and / or "genetic modification" and / or "expansion" and / or "combination" and / or "filling" and / or "washing" and / or "separation" and / or that the process step comprises the process steps "enrichment" and / or "selection" and / or "activation" and / or "loading" and / or "genetic modification" and / or "expansion" and / or "combination" and / or "filling" and / or "washing" and / or "separation".

[0083] Details of these process steps are provided below. Any description given with respect to any feature or method step described herein may apply to any one, any combination, or all of the specified bioprocess steps and / or bioprocess operations. One advantage of the proposed system is that it can be adapted to different process steps as needed. A further advantage of the proposed system is that the adaptation to different process steps can be done while the bioprocess is already running. In particular, the order of process steps can be changed and / or process steps can be repeated.

[0084] Here, preferably, the processing steps "enrichment" and / or "selection" and / or "activation" and / or "loading" and / or "genetic modification" and / or "expansion" and / or "combination" and / or "filling" and / or "washing" and / or "separation" can be performed automatically and / or repeated in any order as part of one bioprocess without manual intervention, by transferring the cell culture between cartridges 4 as needed. In any case, here, preferably, at least one transfer of the cell culture from one cartridge 4 to another cartridge 4 can be performed fully automatically.

[0085] Considering cartridge 4 from a general perspective, bioprocessing system 1 may include a single-step cartridge 4, whereby bioprocessing system 1 performs only a single processing step operation within single-step cartridge 4. The fewer processing steps combined within cartridge 4, the easier it is to flexibly combine and repeat steps.

[0086] Additionally or alternatively, the bioprocessing system 1 may include a common cartridge 4, whereby the bioprocessing system 1 performs processing steps, particularly consecutive processing step operations, within the common cartridge 4. This reduces the number of transfer steps between operations.

[0087] The processing steps "enrichment" and / or "selection" and / or "activation" and / or "loading" and / or "genetic modification" and / or "combination" and / or "filling" and / or "washing" and / or "separation" operations can be performed within the cartridge 4 in a single step.

[0088] As already alluded to, a pre-configured cartridge 4 may comprise a pre-configured cartridge fluidic structure 5, and / or a pre-configured cartridge 4 may comprise specifically pre-configured functional devices 13 for the operations to be performed inside the cartridge 4, and / or at least some, preferably all, of the cartridges 4 used for the operations may be identical prior to pre-configuration. A pre-configured functional device 13 is a functional device 13 that has been added to the cartridge as part of pre-configuration. The functional device 13 itself may have a standard configuration.

[0089] The cartridge 4, with no pre-configured contents, can be viewed as an adapter between the base structure 3 that provides the energy and tissue, and the functional devices 13 and fluidic structures that together perform the actual bioprocess. This layer of abstraction allows for commonality at the level of the base structure and flexibility at the level of the cartridge.

[0090] As already explained in terms of the preferred embodiment of the unit operation station 2, the standardized base structure interface 7 may comprise an active energy transmission interface, and the base structure 3 may transmit mechanical and / or pneumatic and / or electrical energy to the cartridge 4 via the active base structure interface 7 and the matching cartridge interface 8, and in particular drive the functional device 13 of the cartridge 4 by means of the transmitted energy. Here, preferably, the interface comprises a plug system between the base structure 3 and the cartridge 4. The plug may comprise several connectors for electrical energy and signals. Furthermore, a plug for a mechanical drive part, in particular a cartridge drive unit 11, protruding from the base structure 3 may be provided on the cartridge 4.

[0091] Furthermore, the base structure 3 may include a transport and placement interface 35, which here forms the upper portion of the unit operation station 2 together with the overall transport mechanism 17, transfer position 14, etc. The base structure 3 then transports the cartridge 4 via the base structure interface 7 and the matching cartridge interface 8 to a position where the bioprocessing system 1 performs at least one of the operations inside the cartridge 4, and holds the cartridge 4 in that position.

[0092] We have described how the cartridge 4 may include functional devices 13. However, some or all of the functional devices 13 may be multi-use components, and providing a large number of them in the cartridge 4 may not be efficient.

[0093] According to one embodiment, it is proposed that the bioprocessing system 1, in particular the base structure 3, comprises a functional device 13, and preferably the functional device 13 of the base structure 3 is fixed at a predetermined position adjacent to a dedicated standardized interface 6 for one or more operations to be performed by the functional device 13, via a functional interface, in particular a standardized one, of the functional device 13 and a functional receiving interface, in particular a standardized one, of the cartridge 4, preferably standardized for different functional devices 13 for different processing steps, or the functional device 13 of the base structure 3 is transported to a predetermined position adjacent to a non-dedicated standardized interface 6, and the bioprocessing structure performs one or more operations by the functional device 13, via a functional interface, in particular a standardized one, of the functional device 13 and a functional receiving interface, in particular a standardized one, of the cartridge 4, preferably standardized for different functional devices 13 for different processing steps.

[0094] The functional interface may be a dedicated interface, if necessary. Every cartridge 4 may have one or more dedicated interfaces, which may not always be used depending on the application of the cartridge 4. However, a cartridge 4 may also have one or more standardized functional interfaces that are not dedicated to a single operation or process step. For example, different process steps may involve applying contactless energy to dedicated cartridge fluidic structures 5 and / or functional devices 13. The cartridge fluidic structures 5 and / or functional devices 13 are arranged in standardized locations on the cartridge 4 and may include additional components, such as reflectors not yet described, or a contactless energy emitting device that can receive contactless energy in a standardized manner through an interface, which may be a hole of a certain dimension in the cartridge 4 that is in close contact with or adjacent to the individual cartridge fluidic structures 5 and / or functional devices 13. The cartridge fluidic structures 5 may be single-use or may include a hard tube or the like that can connect to the functional device 13 and receive contactless energy. The standardized functional interface may be a standardized interface 6.

[0095] As shown in Fig. 10b), the functional device 13 can be provided in a standardized container 36 for interaction with the cartridge 4 and / or base structure 3, preferably such that the base structure 3 does not need to know what the contents are in order to handle the container 36. The container 36 and / or receptacle 15 and / or cartridge 4 can be of a standard size and / or have a standard transport interface. The container 36 and / or receptacle 15 can have a standard interface 6 for connecting to the cartridge 4 and / or base structure 3.

[0096] All functional devices 13 or combinations of functional devices 13 of the base structure 3 not specifically mentioned yet may be fixed or placed inside the container 36 .

[0097] According to one embodiment, it is proposed that the bioprocessing system 1 performs operations inside the cartridges 4, with identical interfaces between the cartridges 4 for receiving the cell culture, in particular from the receptacle liquid container 56, and / or for delivering the cell culture from the cartridge 4 after the operation, in particular to the receptacle liquid container 56 of the receptacle 15, and / or for receiving consumables, in particular from the receptacle 15. In Figures 2 and 3 it is clear how a single type of interface can be used to connect the cartridges 4 to different receptacles 15. It is also clear that, although not shown, instead of the receptacle 15 in Figure 3 another cartridge 4 can be placed next to the illustrated cartridge 4. In this case too, the same interface can be used for transferring fluids between the receptacle 15 and / or the cartridges 4. Here, the interface between the cartridge 4 and the receptacle 15 comprises a single tubing connection. The interface between two cartridges 4 comprises one or two tubing connections here. It is also clear from FIG. 3 that a receptacle 15 connected to a cartridge 4 can be further connected to other receptacles 15, which may be connected to the same cartridge 4, different cartridges 4, or no cartridge at all.

[0098] In another embodiment, the base structure 3 and / or the receptacle 15 and / or the cartridge 4 may include a pump to pump liquid through the interface. In particular, the pump may be part of the base structure 3 or may be provided by the base structure 3 for the interface between cartridge 4 and cartridge 4, cartridge 4 and receptacle 15, or receptacle 15 and receptacle 15. In this way, a certain number of pumps can be used in the base structure 3 as a multi-use component with various cartridges 4 and receptacles 15. There may be some standard pumps for the interface and other pumps in the cartridge 4 as needed. In one embodiment, the pump is part of the base structure 3 and the standardized base structure interface 7. The pump may protrude into the cartridge 4 when connected or may otherwise act on the cartridge fluidic structure 5.

[0099] Additionally or alternatively, the bioprocessing system 1 may perform operations within the cartridge 4 with the same interface between the cartridges 4 and / or for receiving mechanical and / or pneumatic and / or electrical energy and / or cable-coupled signals from the base structure 3 and / or for receiving energy from the functional device 13 and / or for supplying cell culture to the functional device 13.

[0100] Generally, the cartridge 4 may comprise one or more fluid interfaces 37. Preferably, one or more interfaces for receiving the cell culture and / or for delivering the cell culture and / or for receiving consumables and / or for supplying the cell culture to the functional device 13 are the same fluid interface 37.

[0101] The consumable may be a liquid, a bag containing a liquid, a filter, or the like.

[0102] This interface description may apply to any one, any combination, or all of the different cartridges 4 and / or receptacles 15 described herein. Preferably, at least 50% of the cartridges 4 and / or cartridge 4 types will have a standardized interface 6 having one or more of the described features.

[0103] The term "identical" always means functionally identical. Of course, the tubes may for example be arranged slightly differently.

[0104] The preferred method for connecting the tubes of the different cartridges 4, receptacles 15 and / or functional elements is tube welding, as described below. Other connection methods, such as sterile connectors, can also be used. Preferably, at least some, and preferably all, fluid interfaces 37 are used in defined positions relative to the base structure 3 so that the base structure 3 can interact with the interfaces and, in particular, connect the tubes in a standardized manner.

[0105] Preferably, the cartridge 4 and / or receptacle 15 herein includes up to four inlets and outlets across all of its fluid interfaces. In particular, each fluid interface 37 includes exactly one potential fluid connection that can be used as an inlet or an outlet. For any one, any combination, or all of the one or more preconfigured cartridges 4 described, at least one fluid interface can be used as an inlet, and / or at least one fluid interface can be used as an outlet, and / or at least one fluid interface can be used consecutively for two different connections, particularly to two different receptacles 15 and / or cartridges 4. For any one, any combination, or all of the one or more preconfigured cartridges 4 described, one fluid interface can be used as the only inlet, and / or one fluid interface can be used as the only outlet, and / or one fluid interface can be used as the product outlet and one fluid interface can be used as the only waste outlet, and / or exactly two fluid interfaces can be used as inlets. The product outlet is the outlet for transferring the cell culture from the cartridge 4.

[0106] Here, preferably, the fluidic interface 37 comprises a section of tubing which is likewise arranged inside the cartridge 4 and which is directly connected to the functional device 13 of the cartridge 4. Here, preferably, the internal fluidic structure of the cartridge 4 is not changed after its pre-configuration. In this case, the cartridge fluidic structure 5 is changed only by connecting the interface to a different element, tubing, etc. outside the cartridge 4.

[0107] According to one embodiment, it is proposed that the bioprocessing system 1 comprises a receptacle 15, which is pre-configured with a receptacle fluid structure 16 for containing and supplying and / or receiving liquids, cell cultures and / or consumables that the bioprocessing system 1 uses in at least part of its operations, and preferably the receptacle 15 comprises a fluid interface 37 identical to the fluid interface 37 of the cartridge 4.

[0108] The receptacle 15 may be the primary method of transporting liquids within the bioprocessing system 1. In particular, cell cultures can be transported from the receptacle 15 to the cartridge 4 prior to one or more operations performed within the cartridge 4 and / or from the cartridge 4 to the receptacle 15 after one or more operations performed within the cartridge 4. The receptacle 15 may also be the only method of transporting cell cultures. However, the receptacle 15 may also be used for, or even exclusively for, the transport of consumables that are or contain liquids, such as waste products, media, buffers, etc. Preferably, the receptacle 15, and in particular the receptacle liquid container 56, can be filled at a media filling service station 55 of the bioprocessing system 1. The media filling service station 55 may include one or more tanks containing liquids, such as media or buffers, for many bioprocesses. This makes the introduction of these liquids into the bioprocessing system 1 much easier, since the liquid does not need to be added in small amounts each time it is needed.

[0109] The following describes in detail preferred cartridges 4 that can be used in the bioprocessing system 1. For any one, any combination, or all of the cartridges 4 described herein, only the operation of each of the described processing steps (single-step cartridges 4) or multiple processing steps (common cartridges 4) may be performed within the cartridge 4, or only the operation of one or two other processing steps may be performed within the cartridge 4. Furthermore, any combination of the described cartridges 4 may be used in the bioprocessing system 1 for a single bioprocess and / or different bioprocesses performed in parallel. Nevertheless, preferably, one cartridge 4 is used for only one bioprocess.

[0110] According to one embodiment, it is proposed that the bioprocessing system performs one or more or all operations of the processing step "concentration" and / or the processing step "washing" and / or the processing step "separation" via a base structure 3 inside a pre-configured centrifuge cartridge 39 having a centrifuge chamber 40 as a functional device 13 inside the centrifuge cartridge 39.

[0111] The centrifuge cartridge 39, exemplarily shown in FIG. 9, can be preconfigured by disposing the centrifuge chambers 40 therein, particularly by disposing the centrifuge chambers 40 in predetermined positions. Some or all of the cartridges 4 may have such predetermined positions. The predetermined positions may be dedicated positions for a centrifuge or another functional device 13, or may be predetermined positions for functional devices 13 of a certain size. For example, the predetermined positions may include connecting elements such as screws or screw holes, clip elements, etc. The cartridge 4 may also include multiple overlapping predetermined positions for functional elements of different sizes, whereby functional elements of different sizes preferably utilize different combinations of connecting elements depending on their size and / or position. For example, the floor of the cartridge 4 may be designed as a plug-in board.

[0112] The centrifuge chamber 40 may be a fluidized bed centrifuge chamber 40 .

[0113] In general, as indicated by the use of "and / or," it should be understood that different cartridges 4 of the same type for differently performed processing steps can be used in a single bioprocessing system 1, for example, different pre-configured cartridges 4.

[0114] Hereinafter, preferred configurations of inlets and outlets are described, with all inlets and / or outlets being part of the fluidic interface 37. Of course, the definitions of inlets and outlets are valid for each use of the pre-configured cartridge 4 and may be modified in different configurations.

[0115] Preferably, the centrifuge cartridge 39 has exactly one dedicated inlet for the cell culture and / or exactly one or exactly two dedicated inlets for the buffer. Alternatively, the centrifuge cartridge 39 may have exactly one inlet specifically for the cell culture and the buffer. The centrifuge cartridge 39 may have exactly one waste outlet and / or exactly one product outlet. The inlets and / or outlets may be directly connected to the centrifuge chamber 40.

[0116] The centrifuge cartridge 39 may further comprise one or more, particularly single-use, flow sensors and / or pumps.

[0117] The cartridge interface 8 of the centrifuge cartridge 39 may comprise and use connections for mechanical and / or electrical energy, and / or for the centrifuge chamber 40, and / or for control signals from the flow sensor and / or for the pump. A centrifuge control chip that receives sensor signals from the flow sensor and controls the centrifuge may be located inside the centrifuge cartridge 39 and may be powered via the cartridge interface 8.

[0118] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "concentration" and / or the processing step "separation" via a base structure 3 inside a preconfigured acoustic cartridge 41 having at least one acoustic wave generator, in particular a piezoelectric element, as a functional device 13 inside the acoustic cartridge 41.

[0119] The acoustic cartridge 41, exemplarily shown in FIG. 10 as an exemplary cartridge 4 with a functional interface, can be pre-configured by placing an acoustic wave generator inside the acoustic cartridge 41, particularly by placing the acoustic wave generator in a predetermined position. The acoustic wave generator may include one or two transducers, particularly interdigital transducers. The acoustic wave generator may further include an acoustic reflector. The acoustic cartridge 41 may be identical to the centrifuge cartridge 39 prior to pre-configuration. The acoustic wave generator, and thus the reflector, may be housed in a modular mini-cartridge 42 that can be placed inside the acoustic cartridge 41 and preferably clipped to the acoustic cartridge 41. The acoustic wave generator may include a tube or the like and may be part of the cartridge fluidic structure 5. Alternatively, the tube or the like may be inserted into the mini-cartridge 42 during pre-configuration.

[0120] FIG. 9 also shows how the centrifuge cartridge 39 can have a different functional interface than the acoustic cartridge 41, even though the functional interface of the centrifuge cartridge 39 is not used there.

[0121] Acoustic cell separation works by applying acoustic waves to the cartridge fluidic structure 5, generating standing waves. The cell culture is forced to flow with a sheath flow buffer through the fluidic structure by the standing waves, and cells are separated into different passages of the fluidic structure based on their size.

[0122] Preferably, the acoustic cartridge 41 comprises exactly one dedicated inlet for the cell culture and / or exactly one dedicated inlet for the buffer. A continuous flow of sheath flow may be supplied, particularly from the receptacle 15, via the buffer inlet during concentration or separation. The acoustic cartridge 41 may also comprise exactly one waste outlet and / or exactly one product outlet. The inlets and / or outlets may be directly connected to the part of the cartridge fluidic structure 5 where the acoustic waves of the acoustic wave generator are applied.

[0123] The acoustic cartridge 41 may further comprise one or more, particularly single-use, flow sensors and / or pumps and / or bubble sensors and / or temperature sensors for the acoustic wave generator and / or cell culture. The acoustic cartridge 41 may also comprise a cooling device for the acoustic wave generator, which may be active or passive. If the cooling device is an active cooling device, it may be powered via a standardized interface 6. The cooling device may comprise one or more cooling elements, such as cooling ribs and / or ventilation devices. The cooling device may be part of the acoustic cartridge 41 prior to pre-configuration or may be added to the acoustic cartridge 41 as part of pre-configuration.

[0124] The cartridge interface 8 of the acoustic cartridge 41 may comprise and use connections for electrical energy and / or for control signals for the pump and / or acoustic wave generator and / or from the flow sensor and / or from the temperature sensor. An acoustic control chip that receives sensor signals from the flow sensor and / or bubble sensor and / or temperature sensor and controls the acoustic wave generator and / or pump may be located inside the centrifuge cartridge 39 and may be powered via the cartridge interface 8. If the temperature of the acoustic wave generator or the cell culture reaches a predetermined threshold, the control chip may shut off the acoustic wave generator.

[0125] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "concentration" and / or the processing step "separation" via a base structure 3 inside a preconfigured acoustic cartridge 41 having at least one acoustic wave generator as a functional device 13 outside the acoustic cartridge 41.

[0126] By locating the acoustic wave generator outside the cartridge 4, the acoustic wave generator can be easily reused. Preferably, the acoustic wave generator is capable of applying acoustic waves to the cartridge fluid structure 5 of the acoustic cartridge 41, in particular to the flow path of the cartridge fluid structure 5, or comprises an automatically replaceable and / or sterilizable tube or the like that forms the flow path and is connectable to the cartridge fluid structure 5 of the acoustic cartridge 41.

[0127] The acoustic cartridge 41 may comprise a functional interface for an acoustic wave generator, a fluidic structure to which acoustic waves are applied from one or more sides via the functional interface, and preferably a reflector for the acoustic waves. Part of the cartridge fluidic structure 5 may comprise, as part of the functional interface, a tube or the like for applying acoustic waves in a defined manner at a defined location, in particular at an end of the acoustic cartridge 41. The acoustic wave generator can be docked to the functional interface. The functional interface may comprise a positioning element 43 that ensures a precise fit between the cartridge 41 and the acoustic wave generator. A reflector may also be part of the acoustic cartridge 41.

[0128] The functional device 13 can be placed in a standard container 36 with standard connections to cartridges 4 for various techniques such as acoustic waves, light energy applications, electromagnetic fields, etc.

[0129] The cartridge 4 may have a standard slot for an adapted mini-cartridge 42, which is provided with fluid structures adapted for energy utilization, possibly reflectors etc., and which can be placed in a standard position on the cartridge 4, for example via a clip mechanism (Fig. 10a).

[0130] Any or all of the flow paths within the acoustic cartridge 41 can be fitted with a monitoring system during use. This monitoring system can be used to regulate the flow of liquid through the paths, for example, by changing the pump speed. The monitoring system can also be used to detect defects in the paths, such as blockages, air bubbles, or manufacturing issues such as narrow passages or improper acoustic coupling. For example, the monitoring system can be an optical microscope focused on the paths and, for example, under strobe light or using low light exposure, can detect the presence and size of cells passing through the paths.

[0131] The monitoring system may be part of the acoustic cartridge 41. Alternatively, a functional interface between the acoustic cartridge 41 and the acoustic wave generator may be used for the monitoring system, in which case the monitoring system may be part of the base structure and / or part of the vessel 36 that includes the acoustic wave generator.

[0132] The same concepts can be applied to other operations in the same or other process steps, particularly separation and / or selection process steps, to detect and / or regulate the correct functioning of these operations. Accordingly, one or more standard vessels 36 may generally be equipped with a monitoring system.

[0133] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "concentration" and / or the processing step "separation" via a base structure 3 inside a pre-configured DLD cartridge 4 having a deterministic lateral displacement chamber as a functional device 13 inside the DLD cartridge 4.

[0134] The DLD cartridge 4 may be pre-configured by disposing a deterministic lateral displacement chamber inside the DLD cartridge 4, and in particular by disposing the deterministic lateral displacement chamber in a predetermined position.

[0135] Preferably, the DLD cartridge 4 has exactly one dedicated inlet for the cell culture and / or exactly one dedicated inlet for the buffer. Alternatively, the DLD cartridge 4 may have exactly one inlet specifically for the cell culture and the buffer. The DLD cartridge 4 may have exactly one waste outlet and / or exactly one product outlet. The inlet and / or outlet can be directly connected to the deterministic lateral displacement chamber.

[0136] The DLD cartridge 4 may further include one or more, particularly single-use, flow sensors. The DLD cartridge 4 may include valves for starting and stopping the flow of buffer and / or for starting and stopping the flow of cell culture. Additionally or alternatively, the DLD cartridge 4 may include at least one pump for controlling the flow of buffer and / or cell culture.

[0137] In general, liquid conveying means other than a pump can be used to induce liquid in the tube.

[0138] The cartridge interface 8 of the DLD cartridge 4 includes and can be used to drive fluid and / or actuate valves and / or control control signals for the valves and / or from flow sensors. It should be understood that the exemplary description of the cartridge 4 is not limiting, and that, for example, the DLD cartridge 4 can also include electrically actuated valves and thus electrical connections for actuating the valves.

[0139] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the process step "enrichment" and / or the process step "separation" via a base structure 3 inside a preconfigured electric and / or magnetic separation cartridge 4 having at least one sorting element, in particular an electric field and / or magnetic field generating element, as a functional device 13 inside the magnetic separation cartridge 4.

[0140] Preferably, the electric and / or magnetic separation cartridge 4 is a flow cytometry cartridge 4, which comprises at least one laser and detection optics as functional devices 13 inside the flow cytometry cartridge 4, and at least one, preferably at least eight, or at least 16 sorting elements, in particular charged plates.

[0141] Alternatively, the electrical and / or magnetic separation cartridge 4 may be a negative magnetic immunoadhesion cartridge 4 and may comprise at least one magnetic field generating element inside the cartridge 4 .

[0142] Cell labeling for electric and / or magnetic separation can occur in the incubator service station 38 of the bioprocessing system 1 or in the electric and / or magnetic separation cartridge 4. Cell labeling can occur in the electric and / or magnetic separation cartridge 4, after which the cell culture can be transferred to the incubator position 52. Transfer can occur by transferring the electric and / or magnetic separation cartridge 4 or by transferring the cell culture to the receptacle 15 and later returning it.

[0143] Preferably, the electric and / or magnetic separation cartridge 4 comprises exactly one dedicated inlet for the cell culture and / or exactly one dedicated inlet for the buffer. A continuous flow of sheath flow may be provided during concentration or separation, particularly from the receptacle 15, via the buffer inlet. The electric and / or magnetic separation cartridge 4 may also comprise exactly one waste outlet and / or exactly one product outlet. The inlets and / or outlets may be directly connected to the part of the cartridge fluidic structure 5 where the electric and / or magnetic separation energy is applied.

[0144] The cartridge interface 8 of the electric and / or magnetic separation cartridge 4 may comprise and use connections for mechanical and / or electrical energy and / or control signals for the laser and / or detection optics and / or sorting elements and / or magnetic field generating elements. An electric and / or magnetic separation control chip may be located inside the centrifuge cartridge 39 and may be powered via the cartridge interface 8.

[0145] According to one embodiment, the bioprocessing system 1 performs one or more or all of the operations of the processing step "enrichment" and / or the processing step "separation" via a base structure 3 inside a preconfigured electric and / or magnetic separation cartridge 4 having at least one sorting element, in particular an electric field and / or magnetic field generating element, as a functional device 13 outside the cartridge 4, preferably the electric and / or magnetic separation cartridge 4 is a flow cytometry cartridge 4 and the bioprocessing system 1, in particular the base structure 3, comprises at least one laser and detection optics and at least one sorting element, in particular a charged plate, as a functional device 13 outside the flow cytometry cartridge 4, or the electric and / or magnetic separation cartridge 4 is a negative magnetic immunoadhesion cartridge 4 and the bioprocessing system 1, in particular the base structure 3, comprises at least one magnetic field generating element outside the electric and / or magnetic separation cartridge 4.

[0146] By locating the sorting element outside the cartridge 4, the sorting element can be easily reused. Preferably, the sorting element comprises an automatically replaceable and / or sterilizable tube or the like that can apply an electric and / or magnetic field to the cartridge fluidic structure 5 of the electric and / or magnetic separation type cartridge 4 or that can be connected to the cartridge fluidic structure 5 of the electric and / or magnetic separation type cartridge 4.

[0147] The cartridge 4 of the electric and / or magnetic separation type may comprise a functional interface for the sorting element. Part of the cartridge fluidic structure 5 may comprise, as part of the functional interface, tubes or the like for applying an electric and / or magnetic field in a defined manner and in a defined position, in particular at the end of the cartridge 4 of the electric and / or magnetic separation type. The sorting element can be docked to the functional interface. The functional interface may comprise positioning elements 43 that ensure a precise fit between the cartridge 4 and the sorting element. A laser can also be located outside the cartridge 4.

[0148] The cartridge 4 may have a standard slot for an adapted minicartridge 42 with a fluidic structure adapted for the application of an electric and / or magnetic field.

[0149] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all of the operations of the process step "concentration" and / or the process step "washing" and / or the process step "separation" via a base structure 3 inside a pre-configured filter cartridge 44 having at least one filter 45 as a functional device 13 inside the cartridge 4. The filter cartridge 44 is also shown in Figure 9. The illustrated cartridge 4 is exemplary of other possible cartridges 4.

[0150] Preferably, filter cartridge 44 has exactly one dedicated inlet for the cell culture and / or exactly one dedicated inlet for the buffer. Filter cartridge 44 may also have exactly one waste outlet and / or exactly one product outlet. The inlets and / or outlets may be directly connected to filter 45.

[0151] The filter cartridge 44 may further comprise one or more, particularly single-use, flow sensors and / or pumps, particularly for generating transmembrane pressure across the filter 45 .

[0152] The cartridge interface 8 of the filter cartridge 44 may provide and use connections for electrical energy, and / or for the pump, and / or for control signals from the flow sensor. A control chip for the filter 45 that receives sensor signals from the flow sensor and controls the pump may be located inside the filter cartridge 44 and may be powered via the cartridge interface 8.

[0153] Here, the filter 45 is preferably pre-configured as part of the cartridge fluid structure 5 .

[0154] According to one embodiment, it is proposed that the bioprocessing system 1 automatically connects the centrifugation cartridge 39 and / or the acoustic cartridge 41 and / or the DLD cartridge 4 and / or the electrical and / or magnetic separation cartridge 4 and / or the filter cartridge 44, in particular via one of the fluidic interfaces 37, to the cartridge 4 or receptacle 15 containing the cell culture before the processing step and / or the receptacle 15 containing the buffer as a consumable and / or the receptacle 15 for receiving the washed or separated or concentrated portion of the cell culture after the processing step and / or the receptacle 15 for receiving the waste portion of the cell culture and / or the consumable, preferably the cell culture and the buffer are supplied via the same fluidic interface 37. A tube connection system 23, in particular a tube welding system 24, can be used for any of these connections.

[0155] Here, preferably, the centrifugation chamber 40 and / or the deterministic lateral displacement chamber and / or the filter 45 are single-use components for the bioprocessing system 1, i.e., they can be sterilized, but for that they need to be removed from the bioprocessing system 1. The bioprocess may include two or more of the processing steps "concentration", "separation" and "washing", and in particular the bioprocess may include at least two processing steps "washing".

[0156] In the bioprocessing system 1, one centrifuge cartridge 39 and / or DLD cartridge 4 may be reused for multiple processing steps of one and the same bioprocess, with or without processing steps "cleaning" and optionally disinfection steps in between.

[0157] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "selection" via a base structure 3 inside a preconfigured magnetic selection cartridge 54 having at least one magnetic field generating element as a functional device 13 inside or outside the cartridge 4, preferably the magnetic selection cartridge 54 is an electric and / or magnetic separation type cartridge 4, or the electric and / or magnetic separation type cartridge 4 and the magnetic selection cartridge 54 are combined into a common cartridge 4, more preferably the electric and / or magnetic separation type cartridge 4 is a magnetic separation type cartridge 4 that shares one or more functional devices 13 with the magnetic selection cartridge 54 inside the common cartridge 4.

[0158] The shared functional device 13 may act on the same part of the cartridge fluidic structure 5 or on different parts of the cartridge fluidic structure 5 for different processing steps. Both processing steps may be separate processing steps, and the product may be removed from the cartridge 4 in between, for example for coating or incubation.

[0159] All the explanations given regarding the functional elements of the electrically and / or magnetically separated cartridge 4 may also apply here.

[0160] Magnetic selection is preferably performed on cell cultures incubated with magnetic beads coated with antibodies, in particular antibodies targeting CD62L and / or CD4 and / or CD8 and / or CD56 and / or CD3.

[0161] Instead of, or in addition to, selecting target cells by magnet selection, unwanted cells may be selected by magnetic selection.

[0162] Preferably, the magnetic selection cartridge 54 comprises exactly one dedicated inlet for the cell culture, in particular the cell culture incubated with magnetic beads, and / or exactly one dedicated inlet for the liquid containing the magnetic beads. The magnetic selection cartridge 54 may also comprise exactly one waste outlet and / or exactly one product outlet. The inlets and / or outlets can be directly connected to the functional device 13.

[0163] The cartridge interface 8 of the magnetic selection cartridge 54 may include and use connections for electrical energy and / or control signals for the magnetic field generating elements. A magnetic field generating element control chip may be located inside the magnetic selection cartridge 54 and may be powered via the cartridge interface 8.

[0164] The preferred magnetic selection is performed by mixing the cell culture with the magnetic beads, for example, in the magnetic selection cartridge 54, or in the mixing cartridge, or in the incubator service station 38, then incubating the cell culture, then transferring the cell culture to the magnetic selection cartridge 54 as needed, then flowing the cell culture into the column of the magnetic selection cartridge 54, then removing or deactivating the magnetic field generating element, particularly by removing the container containing the magnetic field generating element or by stopping the electromagnet, then flowing the buffer, particularly from the inlet of the magnetic selection cartridge 54, into the column, and capturing the remainder as the cell culture, particularly after the outlet of the magnetic selection cartridge 54.

[0165] After selection, the waste can be transferred from the magnetic selection cartridge 54 or any other yet undescribed selection cartridge 4 to a storage location, in particular inside a receptacle 15. The same may be true for cartridges 4 in which concentration and / or washing and / or separation takes place. In this way, the stored receptacle can still be used, for example, to repeat the "selection" process.

[0166] If criteria such as the number or viability of the target cells are not met, the waste material may be subjected to another processing step, in particular a selection step, to obtain more target cells. This allows the bioprocess to be flexibly modified to include more steps, with the waste material becoming a cell culture to obtain more target cells. The further step of combining both portions of the target cells can be performed automatically. Alternatively, the waste material can be subjected to the same processing step and the steps repeated.

[0167] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "selection" via a base structure 3 inside a pre-configured buoyancy selection cartridge 4 having at least one centrifuge as a functional device 13 inside or outside the cartridge 4, preferably the buoyancy selection cartridge 4 is a centrifugation cartridge 39 or the centrifugation cartridge 39 and the buoyancy selection cartridge 4 are combined into a common cartridge 4, more preferably the centrifugation cartridge 39 shares a centrifugation chamber 40 with the buoyancy selection cartridge 4 inside the common cartridge 4.

[0168] Buoyancy selection is preferably performed on cell cultures incubated with gas-filled lipid-shell microbubbles coated with antibodies, particularly antibodies targeting CD28 and / or CD3.

[0169] Preferably, the buoyancy selection cartridge 4 comprises exactly one dedicated inlet for the cell culture and / or exactly one dedicated inlet for the liquid containing microbubbles. The buoyancy selection cartridge 4 may also comprise exactly one waste outlet and / or exactly one product outlet. The inlets and / or outlets may be directly connected to the functional device 13.

[0170] The cartridge interface 8 of the buoyancy selection cartridge 4 may include and use connections for electrical energy and / or control signals for the centrifuge. A centrifuge control chip may be located inside the buoyancy selection cartridge 4 and may be powered via the cartridge interface 8.

[0171] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "activation" via a base structure 3 inside a pre-configured activation cartridge 4, preferably having at least one pump as a functional device 13 inside or outside the cartridge 4.

[0172] Activation may include adding liquid, incubation and washing. These steps may be performed in one cartridge 4 or in separate cartridges 4. In some cases activation is not performed in a dedicated activation cartridge 4.

[0173] Activation can involve adding soluble antibodies, or antibody-coated paramagnetic beads, or antibody-containing nanomatrices to the cell culture, which can then be removed by washing.

[0174] The activation cartridge 4 may include a cell count sensor and / or a mixing volume and / or a pump. The cartridge interface 8 of the activation cartridge 4 may include and use connections for electrical energy and / or for the pump and / or for control signals from the cell count sensor.

[0175] Activation can be performed at incubator service station 38. The liquid for activation can be added to activation cartridge 4 and / or at incubator service station 38 and / or at media fill service station 55. Incubation can occur inside or outside of activation cartridge 4.

[0176] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "activation" via an internal base structure 3 of a centrifugation cartridge 39 and / or an acoustic cartridge 41 and / or a DLD cartridge 4 and / or an electrical and / or magnetic separation cartridge 4 and / or a filter cartridge 44 as a common cartridge 4.

[0177] The functional device 13 can be reused for at least the operation of the process step "concentration" or the process step "cleaning" or the process step "separation", as well as for at least the operation of the process step "selection" and / or the operation of the process step "activation". For clarity, one step of concentration, cleaning or separation can be combined with one step of selection or activation. Further steps of other process steps may also be added.

[0178] This common cartridge 4 may comprise a pump and / or an acoustic wave generator and / or a magnetic field generating element and / or a centrifuge as functional device 13 or may be connected to functional device 13 via a functional interface.

[0179] Preferably, activation can be monitored using flow cytometry or microscopy. Activation changes the cell diameter from approximately 6 to 11 microns, which is detectable by transmitted light microscopy of unstained cells. The microscope can be part of each cartridge 4 or can be connected to a container via a functional interface.

[0180] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all operations of the processing step "genetic modification" via a base structure 3 inside a pre-configured genetic modification cartridge 4, and preferably the activation cartridge 4 and the genetic modification cartridge 4 are one common cartridge 4.

[0181] This common cartridge 4 may comprise a pump and / or a centrifuge as functional device 13. Activation and genetic modification may be carried out together with other processing steps, as described for activation, in the common cartridge 4. Activation and / or genetic modification can be carried out by spinoculation inside the cartridge 4, in particular a common cartridge 4 comprising a centrifuge.

[0182] According to one embodiment, the bioprocessing system 1 performs one or more or all operations of the processing step "genetic modification" via an internal base structure 3 of a centrifugation cartridge 39 and / or an acoustic cartridge 41 and / or a DLD cartridge 4 and / or an electric and / or magnetic separation cartridge 4 and / or a filter cartridge 44 as a common cartridge 4, and preferably, it is proposed that the functional device 13 is reused for at least the operations of the processing step "concentration" or the processing step "washing" or the processing step "separation", as well as for at least the operations of the processing step "selection" and / or the processing step "genetic modification".

[0183] Additionally, the bioprocessing system 1 may perform genetic modification via viral transduction, electroporation, or nanoparticle-based delivery. A viral transduction enhancement reagent may be added through an inlet of the genetic modification cartridge 4, particularly from receptacle 15. Alternatively, the viral transduction enhancement reagent may be placed inside the cartridge liquid container 30 during pre-configuration. The viral transduction enhancement reagent may include RetroNectin or a cationic polymer, such as polybrene or a cationic lipid.

[0184] Electroporation involves mixing the cell culture with, preferably, DNA or mRNA in an electroporation buffer. The genetic modification cartridge 4 may have an inlet for DNA or mRNA and / or an inlet for a buffer. Additionally, the genetic modification cartridge 4 may have one or two inlets for a highly conductive sheath medium.

[0185] The genetic modification cartridge 4 may comprise an electroporation cuvette and / or an electric field generating element inside the cartridge 4. The electric field generating elements of electrically isolated cartridges, in particular the charged plates, can be reused, in particular in a common cartridge 4, or the electric field generating elements as part of the base structure 3.

[0186] Here, preferably, the bioprocessing system 1 performs one or more or all operations of the process step "Growth" outside the cartridge 4 and / or in a growth position and / or inside the receptacle 15. Preferably, at least one operation of the process step "Growth" is performed in the incubator service station 38.

[0187] According to one embodiment, it is proposed that the bioprocessing system 1 performs one or more or all of the processing steps "compounding" and "filling" via a base structure 3 inside a pre-configured compounding cartridge 4.

[0188] The compounding cartridge 4 may include an inlet for the cell culture and / or an inlet for the compounding buffer. It may or may not include an outlet for the cell culture, especially if this is the final processing step. The compounding cartridge 4 may include packaging for the final product, or the cell culture may be added to the interior of the compounding cartridge 4.

[0189] According to one embodiment, the bioprocessing system 1 performs one or more or all of the operations of the processing steps "compounding" and "filling" through the internal base structure 3 of a centrifugation cartridge 39 and / or an acoustic cartridge 41 and / or a DLD cartridge 4 and / or an electric and / or magnetic separation cartridge 4 and / or a filter cartridge 44 and / or an activation cartridge 4 and / or a genetic modification cartridge 4 as a common cartridge 4, and preferably it is proposed that the functional device 13 is reused for at least the operation of the processing step "concentration" or the processing step "washing" or the processing step "separation", and at least the operation of the processing step "selection" and / or the operations of the processing steps "compounding" and "filling".

[0190] The bioprocessing system 1 may have dedicated stations for processing steps performed by functional devices 13 external to the cartridge 4, and universal stations for processing steps performed by functional devices 13 internal to the cartridge 4. In the universal stations, different operations of different processing steps may be performed by the bioprocessing system 1 on different cartridges 4 at different times.

[0191] Returning to the general description of the proposed method, in its non-preconfigured state, the cartridge 4 may not yet be available for any processing steps or operations by the bioprocessing system 1 .

[0192] To clarify the present invention, the initial processing steps of the bioprocess for producing genetically modified CAR-T cells from liquid immune cell culture are described below.

[0193] As best seen in Figure 4, a receptacle 15 containing an initial liquid immune cell culture is introduced into the integrated bioprocessing system 1 by an operator at the input / output position 46. From the input / output position 46, the receptacle 15 is transported by the overall transport mechanism 17 to the transfer position 14 of the unit operation station 2. In the illustrated example, the unit operation station 2 is pre-configured for the enrichment processing step by performing a standard routine with each correspondingly pre-configured cartridge 4. In particular, the pre-configured cartridges 4 are transported from the cartridge storage unit 9 to the cartridge drive unit 11.

[0194] In a preferred embodiment, the concentration processing unit operations include at least a countercurrent centrifugation unit operation step. However, as noted above, acoustic separation may be used instead of countercurrent centrifugation. For countercurrent centrifugation, the cartridge fluidic structure 5 of each cartridge 4 comprises a countercurrent centrifugation chamber 40 for receiving the liquid to be centrifuged. Therefore, as noted above, the countercurrent centrifugation chamber 40 is preferably fabricated from a single-use material.

[0195] Before, after, or during transport of the receptacle 15 to the transfer position 14 of the unit operation station 2, the cartridge 4 of the unit operation station 2, which includes a cartridge fluidic structure 5 preconfigured for the concentration processing step, is operably coupled to the cartridge drive unit 11. In particular, the counter-flow centrifugation chamber 40 of the cartridge fluidic structure 5 is operably coupled to a standardized mechanical interface of the cartridge drive structure 33 as part of the operable coupling. This mechanical interface may include a rotor used to rotate the counter-flow centrifugation passage. As mentioned above, the rotor is preferably designed to be used multiple times.

[0196] After a receptacle 15 containing a liquid immune cell culture is transported to the transfer position 14 of the unit operation station 2 and a cartridge 4 is transported to the drive position 10, the receptacle transfer tube 22 and one of the cartridge transfer tubes 21 are connected by welding in a closed connection process (Figure 6a).

[0197] The liquid immune cell culture is then transferred from the receptacle 15 to the counterflow centrifugation element of the cartridge 4 via a welded connection using fluid interface 37 and peristaltic pump 32 to transfer the liquid immune cell culture to centrifugation chamber 40 ( FIG. 3 ). Again, each interface for engaging peristaltic pump 32 is provided by base structure 3, particularly base structure interface 7.

[0198] The liquid immune cell culture is then subjected to countercurrent centrifugation by the countercurrent centrifuge of cartridge 4. To this end, countercurrent centrifugation chamber 40 is fluidly connected to cartridge liquid container 30, which can receive the effluent from the countercurrent centrifugation unit operation step. Alternatively, countercurrent centrifugation chamber 40 is fluidly connected to receptacle 15 provided at transfer position 14, which includes receptacle fluidic structure 16 that does not yet contain any liquid. Receptacle 15 can be transported to transfer position 14 by overall transport mechanism 17 and can be supplied from central supply storage 47.

[0199] It should be noted here that at least a portion of the liquid required at each unit operation station 2 to perform a unit operation may be supplied from the medium filling service station 38 after each liquid is transferred to a receptacle 15 at the medium filling position 48 of the medium filling service station 38 as described above. In this case, the receptacle 15 is transported to the transfer position 14 of a particular unit operation station 2 by the overall transport mechanism 17, and the connection between the cartridge 4 and the receptacle 15 is established in the connection process as described above.

[0200] Optionally, a washing unit operation step may be performed after the countercurrent centrifugation unit operation step (not shown). For this purpose, cartridge 4 may include at least one cartridge fluid reservoir 30 containing a washing fluid. The washing fluid is transferred from cartridge fluid reservoir 30 to the countercurrent centrifugation element. Alternatively, the washing fluid may be provided from media filling service station 38. In the latter case, the washing fluid is transferred from media storage reservoir 36 to fluid reservoir 36 of receptacle 15 as described above. Receptacle 15 is then transported by overall transport mechanism 17 to transfer position 14 of unit operation station 2 and connected to cartridge 4 in the same closed connection process described above.

[0201] Again, the waste liquid from the washing unit operation step is transferred to the cartridge liquid container 30. Alternatively, the waste liquid is transferred to the receptacle liquid container 56 of the receptacle 15 provided at the transfer position 14. Preferably, the same liquid container used to receive the waste liquid from the centrifugation of the liquid immune cell culture is used to receive the waste liquid from the washing unit operation step. However, in alternative embodiments, the cartridge fluidic structure 5 may be equipped with a different cartridge liquid container 30 for receiving the waste liquid and / or may use a different receptacle liquid container 56. After transfer of the liquid waste of the washing unit operation step, the now purified liquid immune cell culture can be eluted from the counterflow centrifugation element and transferred to the second receptacle 15 provided at the second transfer position 14 of the unit operation station 2 (Figure 3). Again, the connection between the receptacle transfer tube 22 of the second receptacle 15 and the cartridge transfer tube 21 of the cartridge 4 is established by a closed connection process. For elution, the cartridge 4 may include a separate cartridge fluid reservoir 30 (not shown) containing an elution fluid such as culture medium. The purified T cells are eluted from the counterflow centrifuge into the receptacle fluid reservoir 56 of the second receptacle 15.

[0202] After all unit operation steps of the enrichment process unit operation have been performed, the welded connections between the receptacle transfer tube 22 and the cartridge transfer tube 21 of each cartridge 4 are severed in a cutting process. The cartridges 4 are then transferred from the cartridge drive unit 11 to the cartridge waste storage unit 49.

[0203] After cutting, the first receptacle 15, now containing the empty receptacle liquid container 56, can be transported by the overall transport mechanism 17 to central waste storage 50 for disposal. The second receptacle 15, now containing the partially processed liquid immune cell culture, is transferred by the overall transport mechanism 17 to a pre-configured unit operation station 2 for a selected processing step.

[0204] During the selection process, certain subtypes of liquid immune cells within the liquid immune cell culture are enriched. In a preferred embodiment, T cell subtypes are enriched. In a further preferred embodiment, T cell subtypes expressing the antigens CD4, CD8, or CD62L are enriched. In a proposed embodiment, the unit operations of the selection process include at least a magnetic separation unit operation step.

[0205] Here, for the selection processing step, a receptacle 15 containing the liquid immune cell culture after the enrichment processing step is transported to a transfer position 14 of a specific unit operation station 2 preconfigured for the selection processing step by a preconfiguration of one of the cartridges 4. As described for the enrichment processing step, the cartridge 4 for the unit operation of the selection processing step, stored in a cartridge storage unit 9 of one of the unit operation stations 2, is moved to a drive position 10 of this specific unit operation station 2.

[0206] After a connection process to establish a fluid connection between receptacle 15 and cartridge 4, a liquid containing magnetic beads coated with antibodies against antigens on the surface of T cells is transferred from cartridge liquid container 30 to receptacle 15 containing the liquid immune cell culture in a liquid addition unit operation step. Again, the fluid connection is established by a welding system 24 that connects the respective transfer tubes in a closed manner.

[0207] As an example, when magnetic beads coated with antibodies against CD4 are added to a liquid immune cell culture, cells in the liquid immune cell culture that contain the CD4 surface antigen will adhere to the magnetic beads.

[0208] In the next step, the receptacle 15 containing the liquid immune cell culture and magnetic beads can be transported to an incubator service station 38 located on a second plane 51. The incubator service station 38 includes at least one incubator position 52 for receiving the receptacle 15. In one embodiment, as shown in FIG. 4, the incubator service station 38 can be designed as a drawer system. Alternatively, the incubator service station 38 can be designed as a shelf system. However, other configurations are also possible. Again, the overall transport mechanism 17 can be used for transport, and an elevator system 53 can be used to transfer the receptacle 15 from the first plane 19 to the second plane 51 (FIG. 4). In the example described here, the incubator service station 38 including at least one incubator position 52 is located on the second plane 51, but it should be noted that it is entirely possible that at least one incubator service station 38 could also be located on the first plane 19.

[0209] In the incubator service station 38, the receptacle 15 is subjected to defined process conditions for a predetermined time in an incubation step. The incubation step is performed to allow binding of the target immune cells (in this case, CD4+ T cells) to the magnetic particles via antigen-antibody binding. Thus, T cells containing a certain antigen corresponding to the antibody coated on the magnetic particles are bound to the magnetic particles.

[0210] After the incubation step, the receptacle 15 is transported from the preconfigured incubator service station 38 back to the unit operation station 2 for the selected processing step.

[0211] Magnetic selection can then be performed on the liquid immune cell culture by the electromagnet or magnet of the magnetic selection cartridge 54. As noted above, the addition of magnetic beads to the liquid immune cell culture may be performed in the magnetic selection cartridge 54, or may be performed in a separate cartridge 4 or in the incubator service station 38 of the bioprocessing system 1.

[0212] A liquid containing CD4+ cells attached to magnetic beads and other cells not attached to magnetic beads is introduced through a first inlet into a channel surrounded by an electromagnet as a functional device 13. While the cells flow through the channel, the electromagnet is switched on, maintaining the magnetic beads and other cells attached to the magnetic beads in the portion of the channel affected by the electromagnet. Thus, cells not attached to magnetic beads exit the channel through a first outlet. The first outlet is fluidically connected to a cartridge liquid container 30 capable of receiving waste liquid from the magnetic selection operation. Alternatively, to receive the waste liquid, the first outlet of the channel is fluidically connected to a receptacle 15 provided at a transfer position 14, which includes a receptacle fluidic structure 16 that does not yet contain liquid. The receptacle 15 may be transported to the transfer position 14 by an overall transport mechanism 17 or may be supplied from a central supply storage 47. Furthermore, a second outlet of the magnetic passageway is fluidly connected to a receptacle 15 provided at a second transfer position 14, the receptacle 15 comprising a receptacle fluidic structure 16 that does not yet contain any liquid. After the immune cell culture has passed through the passageway, the magnet is switched off, allowing all previously bound magnetic beads to be released and leave the field of the electromagnet. Preferably, the magnetic beads are guided to the second receptacle 15. After the target cells (CD4+ cells) are collected in the second receptacle 15, the cells can be separated from the magnetic beads and subjected to an activation treatment step.

[0213] It should be noted here that at least a portion of the liquid required at each unit operation station 2 to perform a unit operation may be supplied from the medium filling service station 38 after each liquid is transferred to a receptacle 15 at the medium filling position 48 of the medium filling service station 38. In this case, the receptacle 15 is transported to the transfer position 14 of the particular unit operation station 2 by the overall transport mechanism 17, and the connection between the cartridge 4 and the receptacle 15 is established in the connection process as described above.

[0214] All connection steps between the cartridge 4 and the receptacle 15 can be performed in a closed connection process as described above.

[0215] According to one embodiment, it is proposed that the cartridges 4 and / or receptacles 15 and / or containers 36 are moved automatically by the bioprocessing system 1, preferably by the same transport mechanism, in particular a robotic manipulator.

[0216] According to one embodiment, it is proposed that the common cartridge 4 has walls that separate the cartridge 4 into compartments for different processing steps, and preferably the functional device 13 is part of the wall and / or the cell culture is transported through a fluidic structure that penetrates the wall.

[0217] As already explained, the electronic process control 28 of the bioprocessing system 1 may repeat a processing step or operation if the cell culture criteria are not met during or after the processing step, thereby allowing for flexible reconfiguration of the bioprocess and / or enabling variations in the processing step to be performed through the same standardized base structure interface 7 by pre-configuring the cartridge 4 differently without reconfiguring the base structure 3.

[0218] As also explained, the standardized interface 6 may not be fully utilized by all cartridges 4 for all process steps, but may be utilized differently for different process steps.

[0219] According to one embodiment, it is proposed that the bioprocessing system 1 can move the standardized interface 6 relative to the rest of the base structure 3 and / or the cartridge 4, or that the standardized interface 6 is immobile.

[0220] The bioprocessing system 1 may include one or more input / output locations 46. These may provide interfaces for the addition of consumables and / or initial cell cultures and / or the removal of packaged cell cultures during use of the bioprocessing system 1.

[0221] In a preferred embodiment, cartridge 4 does not have an energy source or only a battery that does not power one of functional devices 13. Additionally or alternatively, cartridge 4 may be partially or completely made of plastic and / or 3D printed and / or does not include any fluidic structures prior to being pre-configured.

[0222] It should be mentioned that the functional device 13 is not any element having a function, but a device involved in a bioprocess.

[0223] The cartridge 4 may have one wall covering its four sides in particular. The cartridge 4 may have a floor or a cover. The cartridge 4 may be exposed to a defined atmosphere. The cartridge 4 may have a rectangular bottom shape. An unconfigured cartridge 4 may not have any functional elements. Electrical connections may be provided inside the walls and / or lid and / or floor of the cartridge 4.

[0224] Another equally important teaching relates to the cartridge 4 used in the proposed method.

[0225] According to one embodiment, it is proposed that the cartridge 4 is pre-configured with sterile and sealed fluidic structures, and preferably the cartridge 4 is packaged as a ready-to-use component, in particular in a sterile package.

[0226] Another equally important teaching concerns the base structure 3 used in the proposed method.

[0227] Another equally important teaching relates to the bioprocessing system 1 used in the proposed method.

Claims

1. 1. A method for performing a bioprocess on a liquid immune or naive cell culture to obtain a processed cell culture, wherein the processed cell culture is for autologous or allogeneic cell therapy, the bioprocess being carried out in an integrated bioprocessing system (1), the bioprocess comprising a series of processing steps, each of the processing steps comprising at least one operation; 1. A method for performing at least two operations of the bioprocess inside at least two differently preconfigured cartridges (4) by interaction of the base structure (3) with the cartridges (4), the cartridges (4) and the base structure (3) having matching standardized interfaces (6) for interaction between the base structure (3) and each of the cartridges (4), the bioprocess system (1) comprising: a bioprocessing system (1) comprising a base structure (3) and preconfigurable cartridges (4), the bioprocessing system (1) performing at least two operations of the bioprocess inside at least two differently preconfigured cartridges (4) by interaction of the base structure (3) with the cartridges (4) via the same base structure interface (7) and / or a plurality of identical base structure interfaces (7) and matching cartridge interfaces (8).

2. the two operations of the bioprocess are different processing step operations; and / or said bioprocessing system (1) performs at least two operations that are part of different bioprocesses in parallel in the same preconfigured cartridge (4); and / or The bioprocessing system (1) performs at least two operations that are part of different bioprocesses, in particular different processing steps of the different bioprocesses, in parallel in different preconfigured cartridges (4).

2. The method according to claim 1, characterized in that

3. the manipulation comprises one or more of the following processing steps: "enrichment", "selection", "activation", "loading", "genetic modification", "expansion", "combination", "loading", "washing", and / or "separation", and / or The processing steps include the processing steps of "enrichment" and / or "selection" and / or "activation" and / or "loading" and / or "genetic modification" and / or "expansion" and / or "combination" and / or "filling" and / or "washing" and / or "separation".

2. The method according to claim 1, characterized in that

4. 2. The method according to claim 1, characterized in that the bioprocessing system (1) comprises a single-step cartridge (4), whereby the bioprocessing system (1) performs only a single processing step operation inside the single-step cartridge (4), and / or the bioprocessing system (1) comprises a common cartridge (4), whereby the bioprocessing system (1) performs processing steps, in particular consecutive processing step operations, inside the common cartridge (4).

5. 2. The method according to claim 1, characterized in that the preconfigured cartridge (4) comprises a preconfigured cartridge fluid structure (5) and / or the preconfigured cartridge (4) comprises a preconfigured functional device (13) specifically for the operation to be performed inside the cartridge (4) and / or at least some of the cartridges (4) used for the operation are identical before being preconfigured.

6. the standardized base structure interface (7) comprises an active energy transmission interface, and the base structure (3) transmits mechanical and / or pneumatic and / or electrical energy to the cartridge (4) via the active base structure interface (7) and the matching cartridge interface (8), in particular driving functional devices (13) of the cartridge (4) by means of the transmitted energy; and / or The base structure (3) comprises a transport and placement interface (35), and the base structure (3) transports the cartridge (4) to a position where the bioprocessing system (1) performs at least one of the operations inside the cartridge (4) via the base structure interface (7) and the matching cartridge interface (8), and holds the cartridge (4) in the position.

2. The method according to claim 1, characterized in that

7. 2. The method according to claim 1, characterized in that the bioprocessing system (1) performs one or more or all of the operations of the process step "concentration" and / or the process step "washing" and / or the process step "separation" via the base structure (3) inside a preconfigured filter cartridge (44) having at least one filter (45) as a functional device (13) inside the cartridge (4).

8. 2. The method of claim 1, characterized in that the bioprocessing system (1) performs one or more or all operations of the processing step "activation" via the base structure (3) inside a preconfigured activation cartridge (4), preferably having at least one pump as a functional device (13) inside or outside the cartridge (4).

9. The method according to claim 1, characterized in that the bioprocessing system (1) performs one or more or all operations of the processing step "genetic modification" via the base structure (3) inside a preconfigured genetic modification cartridge (4), preferably the activation cartridge (4) and the genetic modification cartridge (4) are one common cartridge (4).

10. The bioprocessing system (1) performs one or more or all of the operations of the processing step "genetic modification" through the base structure (3) inside the filter cartridge (44) as a common cartridge (4), and preferably: The bioprocessing system (1) performs the genetic modification by viral transduction, electroporation, or nanoparticle-based delivery.

10. The method according to claim 9, characterized in that

11. 2. The method according to claim 1, characterized in that the electronic process control (28) of the bioprocessing system (1) repeats a processing step or operation if the criteria of the cell culture are not met during or after the processing step, thereby allowing the bioprocess to be flexibly reconfigured and / or variants of processing steps to be performed via the same standardized base structure interface (7) by pre-configuring the cartridge (4) differently without reconfiguring the base structure (3).

12. 2. The method according to claim 1, characterized in that the cartridge (4) does not have an energy source or has only a battery that does not drive one of the functional devices (13) and / or the cartridge (4) is partially or completely made of plastic and / or 3D printed and / or does not have any fluidic structures before being pre-configured.

13. A cartridge for use in the method according to any one of claims 1 to 12.

14. 14. Cartridge according to claim 13, characterized in that the cartridge (4) is pre-configured with a sterile and sealed fluidic structure, preferably the cartridge (4) is packaged as a ready-to-use component, in particular in a sterile package.

15. A bioprocessing system for use in the method of any one of claims 1 to 12.

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