Bioprocessing Systems

Abstract

1. A bioprocessing system comprising: a series of processing stations for performing operations for a bioprocess; and an automation system comprising: means for manipulating a fluid connection between a first container and a separable second container, thereby creating a sterile connection allowing controlled transfer of fluid or cellular material between the first container and the second container, the means for manipulating the fluid connection being configured to create a sterile connection that can be separated after transfer of fluid or cellular material is completed to be able to operate a further such fluid connection between the first container and a separable third container; and means for controlling the automated series of operations of the processing stations.

Description

[Technical field]

[0001] The present invention relates to bioprocessing systems for manipulating biological samples, and more particularly to automated bioprocessing systems that may be used, for example, to automate cell therapy manufacturing. [Background technology]

[0002] Therapeutics are increasingly using cells rather than small molecules as a starting point. Approaches to manufacturing these products are rapidly evolving to keep up with novel therapeutics that are constantly emerging. In recent years, we have seen an increase in the use of a number of novel classes of cell therapies. One class is autologous cell therapies.

[0003] Autologous cell therapies are a promising class of therapeutics with significant clinical and commercial potential ranging from treating cancer to correcting genetic defects. These therapies involve harvesting cells from a patient, manipulating the cells over a period of days to weeks, and reintroducing the cells into the patient's body to produce a therapeutic effect. The steps performed during autologous cell therapy are often complex; for example, a typical CAR-T process may include a series of steps starting with cryopreserved leukopacks, thawing, washing to remove DMSO, concentrating T cells, activation, transduction, expansion, concentration, finishing the formulation filling into IV bags, and cryopreservation and several other intermediate manufacturing steps. To date, these processes have typically been performed in expensive class 5 clean rooms or isolators with labor-intensive manual processes.

[0004] Due to the complexity of bioprocessing, it is desirable to automate the process while maintaining a closed system that eliminates the need to perform the steps in such high-grade clean rooms, thereby reducing the labor and costs required. A closed system is one in which there is no exposure of the process to the surrounding environment, thereby avoiding contamination of the environment with the product being manufactured, while also eliminating the ingress of contaminants from the environment or cross-contamination from other processes being performed simultaneously. There are systems that have attempted to provide a solution to this, but each has limitations. For example, robotic systems within isolators, where the robots perform standard liquid handling operations, have been developed to perform end-to-end cell therapy processes. However, these systems have no way to minimize the risk of cross-contamination and are limited to performing one patient therapy at a time, making the system expensive and space inefficient. There is also a large burden of assurance to ensure that the isolator can be sterilized and cleaned between each run.

[0005] A more common approach is to use closed systems that include complex consumable elements, which connect the biological sample to all other necessary processing stations, e.g., via tubing fluidly connected to the consumable elements, and provide pumps and valves to perform the steps in a specific sequence. However, these consumable elements are often quite complex to manufacture and install, and as a result are relatively expensive and unreliable. Each consumable element must be individually tailored to the process being performed, making modifications to the system inflexible and expensive to adapt to new processes. As the process becomes more complex, so do these consumables. Furthermore, these systems can usually only operate / operate one consumable element at a time, which makes the bioprocess expensive and space inefficient for expansion to multiple patient use. In many cases, the systems still cannot perform all the steps required for a complete bioprocessing method, and instead often operate multiple independent units in sequence, which means that additional effort and expertise is required to transfer cells (e.g., patient samples) between the independent units. This also introduces an additional risk of cross-contamination, and there is no easy way to detect that contamination is not occurring.

[0006] One method of creating a sterile connection between tubes is tube welding, a process that is performed manually using semi-automated equipment. Sterile tube welding machines are capable of making connections between two tubes with closed ends without exposing the contents of either tube to the environment, and are the only widely accepted means of reusably creating closed connections within a single system. However, existing tube welding systems are generally large and require precise manual operations to properly insert the tube into the welder, properly remove the tube from the welder, and unpinch the weld area to allow for fluid flow after welding. Additionally, existing tube welding systems require visual inspection by the operator after each weld to ensure the success of the weld. Overall, the welding process can take anywhere from four to seven minutes of operator time. As a result of the manual operations, a large portion of the tube is often discarded by the user during each tube welding operation. Furthermore, conventional tube welding systems are not additionally configured to cut the tube and subsequently reliably reseal the ends of the separate tubes, meaning that they may not maintain closure of the contents when the tubes are separated.

[0007] However, there are many reasons why tube welding seems incompatible with full (operator independent) automation. For example, tube welding requires the handling of long flexible tubes that are well known to be extremely difficult to handle by automated handling means such as robots. For example, flexible tubing is unlikely to remain in a well-defined, deterministic position when moved by a robot, making engagement, alignment, and orientation of the tubes difficult. When a sufficiently long flexible tube, restrained at one end, is manipulated at the other end, the path the tube defines will have multiple, non-unique solutions for the path between the restrained and manipulated ends, depending on the internal stresses within the tube. Such systems have many degrees of freedom. Such applications are ideal for humans, but are extremely difficult for automation, and therefore are usually left to human operators. Since the strength of the weld is highly dependent on the precise positioning and compression of the tubes, errors in the engagement of the tubing by the robot can lead to weld failure. It is also difficult to avoid the tubes kinking and / or tangling with other tubes and other parts of the bioprocessing system. Additionally, the tubes could place unexpected tension on the robot or any attached consumables, making the weld less likely to be successful or damaging the connection to the consumable. Finally, while the core welding process is automated, there are no commercially available welding systems that have automated auxiliary processes that require precise manual manipulation, such as unpinning the weld. Thus, until now, it has been considered impractical to develop a bioprocessing system that includes automated tube welding.

[0008] Even if the above challenges could be overcome, there is still significant resistance in the industry to the use of tube welds or other in-process sterile transfers in place of pre-connected single-use tubing sets in systems where reliability and contamination issues are critical, such as bioprocessing systems. In an automated cell therapy process, hundreds of sterile fluid transfers between different containers must be performed per therapy without any of them failing, which could result in contamination of the therapy or the external environment. If single-use tubing sets are used, integrity testing can be performed on the pre-sterilized tubing sets immediately prior to use to ensure that the integrity has not been breached, greatly reducing the risk of loss of sterility / contamination violations. In contrast, in-process connections cannot be pre-checked, and even if each individual transfer or weld has a 99% success rate, when performing 100 sterile fluid transfers in succession, there is only a 35% chance that all sterile fluid transfers will be successful. As a result, to achieve an acceptable 99.9% success rate for 100 welds, the success rate of each individual weld must be 99.999%. One option is to attempt to minimize the number of tube welding operations required by the process, and indeed the ISO 23565 standard for designing equipment systems for cell therapy manufacturing states that "equipment should be designed and utilized such that the number of in-process connections, such as tube welds, is minimized to reduce the risk of contamination." It is also noted that the industry does not currently have any automated, multi-use fluid connectors capable of multiple connection and disconnection cycles.

[0009] Therefore, the industry common sense is to avoid and circumvent the problem of unreliable tube welding altogether by limiting the number of in-process connections through the use of extensive pre-connected disposable consumables, and by performing transfers manually with close operator inspection.

[0010] Thus, none of the conventional approaches are able to provide a flexible, autonomous bioprocessing system that can reliably perform many therapies, and preferably perform multiple therapies simultaneously. Thus, little progress has been made in automating bioprocessing systems that utilize tube welding, due to the significant complexity and size of existing tube welding systems, as well as the stringent requirements for reliability when applied to bioprocessing systems. Summary of the Invention [Problem to be solved by the invention]

[0011] What is needed is a bioprocessing system that can optionally handle multiple patient samples simultaneously and for improved methods of operating sterile fluid connections for closed transfer of fluids and cellular materials, and ideally that can maintain sterility / prevent contamination of consumables and patient samples regardless of whether operations are performed in a sterile or non-sterile atmosphere in such a system. [Means for solving the problem]

[0012] Described herein is a bioprocessing system comprising a series (e.g., a plurality) of processing stations that perform operations for a bioprocess; and an automation system comprising means for manipulating a fluid connection between a first container and a separable second container, thereby creating a sterile connection that allows controlled transfer of fluid or cellular material between the first container and the second container, where the means for manipulating the fluid connection is configured to create a sterile connection that can be separated after transfer of fluid or cellular material is completed to be able to operate a further such fluid connection between the first container and a separable third container; and means for controlling the automated series of operations of the processing stations.

[0013] By providing an automated system that can make (and subsequently disconnect) sterile (fluid) connections between (multiple) different vessels (e.g., an automated system that can connect and / or disconnect vessels aseptically), a series of bioprocessing operations can be performed without the need for a human operator. This eliminates human error and allows the automated system to perform the bioprocessing method with great reliability. As referred to herein, a "vessel" may be considered to be a form of "consumable" (element) in the context of the present invention.

[0014] Advantageously, the sterile connection ensures that transfer of material between the containers can take place without exposing the contents of the containers to the surrounding environment, which could otherwise contaminate the contents. After the transfer of fluid or cellular material between the containers is completed, the containers can be separated and then one / both containers can be fluidly connected to a different container. This means that it is not strictly necessary to provide the automation system (and processing station) within a sterile enclosure.

[0015] The vessel can be relatively simple compared to existing vessels for bioprocessing, since the bioprocessing system (i.e. the operating means) allows both the connection and disconnection of sterile connections.

[0016] Furthermore, bioprocessing systems can be highly flexible since they can be adapted to process a wide variety of bioprocessing methods (i.e., different sequences of bioprocessing operations). Bioprocessing systems are also highly scalable since additional processing stations can be added over time. Thus, such automated systems can be used to process multiple patient samples simultaneously while maintaining separation (e.g., sterility) between different samples, which may be held in one or more containers.

[0017] The means for controlling the automated sequence of operations may be realized by a processing and control unit (e.g., a "control system") of the bioprocessing system. The means for accessing the control system may be local or remote to the control system. The means for controlling the automated sequence of operations of the processing stations may also control the automation system (e.g., the entirety).

[0018] The means for manipulating the fluid connection may be configured to create and / or maintain a closed sterile connection when manipulating (e.g., or creating a sterile connection) the fluid connection between the containers, and to inhibit (and preferably prevent) the contents of the fluidly connected containers from being exposed to the environment. Additionally or alternatively, the means for manipulating the fluid connection may be (further) configured to create and / or maintain a closed sterile separation when separating the fluid connection between the containers, and to inhibit (and preferably prevent) the contents of the separated containers from being exposed to the environment. A closed sterile separation may also be referred to as creating or maintaining a "sterile seal" on (or at) the containers.

[0019] As used herein, the term "closed sterile connection" or "closed sterile separation" preferably includes a sterile connection or separation in which the contents of the container are not exposed to the environment at any stage during the connection / separation process. Thus, a closed sterile connection / separation may be considered to be a "dry" sterile connection / separation, which preferably includes a sterile connection / separation that does not require the use of a sterilant during its formation to ensure it is sterile.

[0020] In fact, for a truly closed connection / separation, no sterilant should be required to prevent contamination. In this way, there is no need to provide and maintain a supply of sterilant (or "disinfectant") within the bioprocessing system, and the need for pumps and valves to transport the sterilant is eliminated. Thus, there is no risk of leakage or spillage of the sterilant that may interfere with electronics or other devices within the bioprocessing system. This also eliminates the risk of the sterilant getting into the container and damaging or killing cells, and it is difficult to completely deliver the sterilant to the desired area while preventing the sterilant from entering the container, thereby damaging or killing cells. Furthermore, this eliminates the need to search for a sterilant that will work with a wide variety of contaminants and completely impregnate the target area. Furthermore, contaminants such as harmful heavy metals or pyrogens can still be harmful and extremely difficult to remove using a sterilant, so it is advantageous to keep the connection closed rather than simply sterile.

[0021] The means for manipulating the fluid connections may be further configured to seal the separated fluid connections such that transfer of fluids or cells to or from the first and second containers is prevented (preferably prevented). In other words, when the fluid connections are separated, each of the separated containers is preferably sealed by the manipulating means and is ready to form a new fluid connection with another container. Thus, the manipulating means may be further configured to create a sterile seal on / to the containers when separating the fluid connections.

[0022] In this way, the vessels can be isolated from the bioprocessing system while maintaining a "closed" system throughout the process of connecting and separating many fluids (e.g. between different vessels). This allows the complete bioprocessing method to be performed without exposure of the vessel contents to the environment during connection or separation, which could potentially cause contamination of the vessel contents and / or the environment. Thus, the vessels can be connected and separated as many times as required for a particular process. Alternatively or additionally, a separate means for sealing (vessels) may be provided. The means for sealing may comprise an electromagnetic source, such as a radio frequency (RF) source.

[0023] The bioprocessing system may further comprise means for placing one or more containers at each of a series of processing stations and for moving the containers between the stations.

[0024] In this way it is possible to carry out some additional steps within the bioprocessing method without the need for operator intervention. The positioning means may be realised by a robotic device which may comprise a robotic arm on the mobile manipulation unit or by a conveyor. The positioning means may also be provided with means for manipulating the fluid connections. Advantageously, the mobile manipulation unit does not have to follow a precise sequence of movements and may carry out asynchronous processes simultaneously.

[0025] The fluid connections may be made by the manipulation means joining together the (free) ends of two (preferably flexible) tubes, each of which can be fluidly connected to a respective container to make a sterile connection between them. The tubes on each container will of course have a length such that the free end is sufficiently spaced from the other end attached to the container to be able to manipulate one or more fluid connections.

[0026] The bioprocessing system may further comprise means for enabling the transfer of fluids or cells between the sterilely connected vessels. The transfer means may be in the form of a pump, such as a peristaltic pump, a syringe pump, and / or a pressure-driven flow pump. Advantageously, the pump may pump fluids or cells between the vessels while maintaining a closed system, thereby preventing contamination of the vessels and / or the surroundings.

[0027] The bioprocessing system may be located in a non-sterile atmosphere, which advantageously significantly reduces the operational costs of the factory and also allows operators to access parts of the bioprocessing system without the risk of contaminating the process.

[0028] The bioprocessing system may further comprise means for determining whether the fluid connections are successfully connected, for example by inspecting the fluid connections, preferably the fluid connections are inspected automatically. The determination may be an inspection, which may be performed mechanically, visually using a machine vision system, etc. The means for determining (e.g. inspection) may be located on a robotic device or elsewhere in the bioprocessing system. The determination may be performed such that if a fault is detected, corrective action may be taken, for example to isolate the contaminated area or to perform a bioburden test. In case of a fault, the determination (e.g. inspection) may be performed before starting fluid transfer through the fluid connections, such that the process may be repeated until a satisfactory connection is made before the process material enters the connection area. Advantageously, this means that the required level of reliability may be maintained even with a large number of in-process connections.

[0029] The bioprocessing system may further comprise an image capture system or device, e.g., a camera with a microscope lens, to inspect the sterile connections and / or to identify each of the vessels. As mentioned above, the sterile connections may be fluid connections made between (preferably flexible) tubes fluidly connected to each vessel, said tubes being joined together by a manipulation means that creates the sterile connections.

[0030] Advantageously, the bioprocessing system can automatically maintain traceability of containers and prevent cross-contamination errors, even when multiple therapeutics are run simultaneously. The bioprocessing system can include multiple such apparatus or devices located at various locations throughout the system. The image capture apparatus or device is sometimes referred to as a machine vision system.

[0031] The bioprocessing system may further comprise one or more sensors configured to detect fluid leakage from the sterile connection. The one or more sensors may comprise at least one of a fluid sensor and a pressure sensor. When the fluid connection between the vessels is made by means of manipulating the free ends of the tubing (e.g., connected to each vessel) to join together, the means of manipulating the fluid connection may further be configured to apply a force (e.g., a tensile force) to the joined tubing on either side of the sterile connection so that mechanical properties may be determined.

[0032] The automated sequence of operations may be controlled according to one or more predefined workflows, preferably one or more reconfigurable bioprocessing workflows. In this way, a specific bioprocessing method may be executed by the bioprocessing system and the process may be easily modified or adapted without requiring any modification of the bioprocessing system itself.

[0033] The means for controlling the automated sequence of actions may be configured to simulate the automated sequence of actions before the bioprocessing system performs said sequence. The means for controlling the automated sequence of actions may communicate at least one result of the simulation to an operator. The at least one result may include an indication of when a particular action occurs, an indication of when a manual step may need to be performed, and / or an indication that a conflict between two simultaneous actions may (or may not) occur. The simulation may use current therapy quality metrics and historical process data to inform the simulation. The simulation may also provide an indication of the likely number of cells of a given type achieved by a given period of time, and may also give an indication of their quality.

[0034] The bioprocessing system may further comprise a monitoring system for verifying that the automated sequence of operations has been performed. The monitoring system may be provided by the means for controlling the automated sequence of operations of the processing stations. Preferably, input to the monitoring system is provided using one or more sensors that are not used by other parts of the bioprocessing system.

[0035] Preferably, the series of processing stations includes means for performing concentration, washing and incubation. In this way, the bioprocessing system can be easily adapted and used to perform all steps of a typical CAR-T process, which require operations to perform lymphocyte concentration, activation, transfection, washing, expansion and harvesting, etc. There may be multiple instances of any of the processing stations, such as to provide redundancy or to reduce the impact of rate-limiting steps. For example, since incubation is usually the rate-limiting step, an additional incubation station may be provided, but it will be understood that any other processing station may be added or removed to reduce the impact of the rate-limiting step.

[0036] Preferably, the series of processing stations includes means for determining cell count, cell viability, and / or cell phenotype. This increases the robustness of the system and allows the operator to track the cell therapy process and determine if it is within specifications, thereby allowing for a more reliable system since out-of-specification processes may be identified early on. In this way, it is possible to automatically monitor the process and determine if the current patient batch being processed is within specifications or not. It also allows for the possibility of adaptive processing, where parameters including cell count are used to update the automated series of operations. Data from the means for determining cell count, cell viability, and / or cell phenotype may be used as input for the means for simulating the automated series of operations.

[0037] The bioprocessing system may be configured to process multiple containers simultaneously, preferably with two or more of the containers containing patient samples. In this manner, multiple bioprocessing methods may be performed in parallel, which may increase the efficiency of processing multiple patient samples. Because the bioprocessing system uses sterile connections to make connections, it is not necessary to sanitize or restart the bioprocessing system to process multiple containers with different patient samples. Containers corresponding to different patient samples may use the same predefined workflow.

[0038] A first container containing a first patient sample may be processed using a different predefined workflow than a second container containing a second patient sample. In this manner, different workflows may be used for different patients without requiring modification of the bioprocessing system or the use of separate bioprocessing systems.

[0039] The means for controlling the automated sequence of actions may be configured to automatically schedule a sequence of actions for the bioprocessing system to follow.

[0040] The sequence of actions may be automatically updated based on input received from at least one sensor of the bioprocessing system. In this way, multiple patient samples may be processed simultaneously by the bioprocessing system while minimizing the risk of conflicts between corresponding bioprocessing workflows. For example, the sequence of actions may be scheduled to minimize and preferably prevent either the processing stations or parts of the automation system from being required simultaneously for separate bioprocessing workflows. If it is not possible to avoid a conflict, the means for controlling the automated sequence of actions may delay one of the conflicting actions based on a preprogrammed or user configurable list of priorities.

[0041] The means for manipulating the fluid connection may be configured to create a sterile connection between a first tube connecting to a first container and a second tube connecting to a second container. Although tubing is extremely difficult to handle by automated handling means, the use of tubing provides several advantages. First, tubing is already widely used in manual bioprocessing, so it is easy to employ and work with third party consumables and other hardware. Second, the overall space and form factor of an automated bioprocessing system can be smaller, as each unit operation only needs to work with small tubing rather than large disposable consumables. Third, because tubing is the primary disposable component and can be manufactured in large quantities, the overall cost is significantly reduced and the overall reliability is increased compared to disposable consumables that contain many components.

[0042] The means for manipulating the fluid connection may comprise a tube welder configured to join a first tube to a second tube. Preferably, the tube welder is configured to join a free end of the first tube to a free end of the second tube, although it will be understood that the connection may be located anywhere along the length of the tubes. As used herein, the term "tube welder" refers to any device configured to join (i.e., weld) a first and second tube (preferably at their free ends), thereby achieving a sterile (fluid) connection between the tubes. Tube welding is a convenient method of creating a sterile connection without the need for a sterilant, i.e., welding is generally accepted to be a "closed (sterile) connection process". The tube welder may further comprise a means for cutting any length of tubing, thereby separating the tubing. The advantage of a sterile connection being a "closed connection" rather than merely being sterilized is that contamination can still enter the connection path, which means that a merely sterilized connection can still be harmful.

[0043] The bioprocessing system may further comprise a means for manipulating the joined tubing to release the pinched portion, thereby establishing a fluid path through the joined tubing. Typically, tube welders do not allow removal of the pinched portion and therefore cannot be easily integrated into a bioprocessing system. Thus, by providing a means for releasing the pinched portion, the bioprocessing method may be performed without operator intervention.

[0044] The bioprocessing system may further comprise a tubing supply means configured to provide auxiliary tubing for use by the means for manipulating the fluid connections.

[0045] One problem with using tubing to form fluid connections between vessels is that the tubing can become tangled with each other and with other parts of the bioprocessing system. It is therefore advantageous to keep the tubing connecting to corresponding vessels relatively short. By providing a tubing supply means, such as a tubing reel, the tubing can be extended as needed while still further minimizing the risk of tangling.

[0046] At least one of the tubes may be provided with at least one identification mark at a position along the tube, preferably the identification mark being readable by an image capture system further configured to determine the position of the identification mark on the tube.

[0047] Preferably, the identification mark is located at a predefined position along said tube. Preferably, the tube comprises a plurality of identification marks, which may be located at regular intervals along its length. The identification mark may indicate the tube size, the tube material, the distance between the identification mark and the corresponding container, the ID of the substance contained in the corresponding container, and / or the orientation of the tube (such as indicating the direction towards the corresponding container). Advantageously, this reduces the risk that the automated system will incorrectly connect two tubes. Additionally, this allows the means for manipulating the fluid connection to engage the first and second tubes in the correct position and orientation, thereby reducing tube waste.

[0048] The bioprocessing system may further comprise at least one tube clip configured to clip (or "hold") the first or second tube in a predetermined fixed / known (or easily identifiable) position relative to the tube clip. The tube clip may be located at a fixed / known position in the bioprocessing system or may be movable to / from a fixed / known position before / after / during a bioprocessing operation. By allowing the tubing to be held in place relative to the tube clip at a fixed / known position, it becomes much easier for automation (e.g., a robotic system) to locate and engage tubing that is not tubing hanging in space. The tube clip may comprise an identification mark that uniquely identifies the position of the tube clip in the bioprocessing system. Advantageously, this allows the means for manipulating the fluid connection to be moved to a precise position in the bioprocessing system to engage the first or second tube. At least one tube clip may comprise additional sensors, such as sensors for detecting the presence of a tube, air in the tube, and / or flow rate of fluid through the tube. The tube clip may also be an active tube clip that can be moved from an open position that retains the tube to a closed position. Advantageously, the tube clip significantly reduces the risk that the automated system will inadvertently engage and manipulate the tube, ultimately improving the reliability of the system.

[0049] The bioprocessing system may further comprise a means for sealing the tube. The means for sealing the tube may comprise an electromagnetic source, such as an RF source.

[0050] Preferably, the bioprocessing system comprises at least one robotic device configured to provide at least one of the following: means for manipulating fluid connections, means for placing one or more containers into each of a series of processing stations, means for enabling transfer of fluids or cells between aseptically connected containers, means for sealing tubing, and / or means for inspecting fluid connections. The at least one robotic device comprises at least one robotic arm and may further comprise at least one end effector. Advantageously, by configuring the robotic device with an end effector in this manner, the robotic device moves towards the tube to be engaged, thereby engaging the tubing near the consumable to be welded, reducing the need for long lengths of flexible tubing. This not only significantly reduces the automation challenges, but also benefits the bioprocessing system overall in terms of reducing the dead volume contained within the tubing.

[0051] Also described herein is an automated system for fluidly connecting two containers (e.g., for use with the bioprocessing system described above), where at least a first container has a tube fluidly connected to a first end, the second end of the tube configured to form a sterile connection with another such tube, the automated system comprising a robotic device (or other automated means) configured to engage the second end of the tube fluidly connected to the first container and position the tube to one or more positions to be manipulated, and means for manipulating a portion of the tube towards the second end of the tube, thereby configuring the second end of the tube to create a sterile connection with another such tube.

[0052] The other tube may itself be connected to a second separate container, or may be a length of tubing that is not connected to a container. For example, this tube may be provided on a tube supply means, such as a tube reel. The means for manipulating the portion of the tube may be part of a robotic device or may be a separate component within an automated system (such as a separate robotic device).

[0053] It will be understood that any of the features described herein in relation to the "automated system for fluidly connecting two vessels" (described above and herein) may be equivalently incorporated into the "bioprocess system" (described above and herein), and vice versa.

[0054] The robotic device may be configured to engage and / or position the tube by moving along one or more predetermined paths.

[0055] Advantageously, by moving along a repeatable and verifiable path, the risk of entanglement between tubes and / or collision with other parts of the automated system is minimized. For example, the robotic device may position a consumable or tube (such as via a tube clip) and navigate in a predetermined manner from that position. In this manner, the starting position may be variable (such as set by various positions of the consumable and tube), and the robotic device engages and positions the tube in a repeatable and verifiable manner from that position, such as by moving along a predetermined vector or set of vectors.

[0056] The means for manipulating the portion of the tube may further comprise means for clamping the portion of the tube towards the second end of the tube, thereby forming a pinched portion on the tube whereby the tube is fluid-sealed upstream of the pinched portion, and means for removing a section of the tube downstream of the pinched portion, thereby removing the second end of the tube, thereby forming a new second end of the tube that has not previously been in contact with another such tube.

[0057] The section of the tubing downstream of the pinched portion may be referred to as the “end section” or “downstream section.” Advantageously, if there is a failure to create a sterile connection by fluidly sealing the tubing upstream of the pinched portion, the process may be repeated until a satisfactory connection is made before process materials enter the connection area.

[0058] The automated system may further comprise a means for enabling controlled transfer of fluid and cellular material between the first and second containers. The means for enabling controlled transfer may be in the form of a pump, such as a peristaltic pump, a syringe pump, or a pressure-driven flow pump. Advantageously, the pump may pump fluid or cells between the containers while maintaining a closed system, thereby preventing contamination of the containers and / or the surroundings. The means for enabling controlled transfer may be configured as an end effector for a robotic arm.

[0059] The means for enabling controlled transfer of fluids and cellular material may further be configured to draw fluid away from the pinched portion within the tube before a sterile connection is made with another such tube. Advantageously, drawing fluid away from the pinched portion may ensure that the pinched portion is dry and improve the reliability of removing the end section of the tube. Additionally, drawing fluid away from the pinched portion may help keep the tube pinched closed during removal of the end section of the tube.

[0060] The means for clamping a portion of the tube may be a station of the bioprocessing system separate from the robotic device. The means for removing a section of the tube may be a station of the bioprocessing system separate from the robotic device. At least one of (i) the means for clamping a portion of the tube and (ii) the means for removing a section of the tube may be configured as an end effector for a robotic arm. In this way, the robotic arm may move to engage the tube, clamp the tube to form a pinched portion, and / or remove an end section of the tube without the need to place the tube in another device. Both the means for clamping and the means for removing may be realized by a single common end effector. Alternatively, the means for clamping and the means for removing may be provided on separate end effectors, separate robotic arms, and / or separate robotic devices. The end effector and / or robotic arm may be the same or different end effectors and robotic arms configured to realize the means for enabling controlled transfer of fluids and cells.

[0061] The means for removing the section of the tube may comprise a cutting blade and at least one of a heating device, e.g., a laser, an RF heater, and an ultrasonic heater, or an inductance heater. In one embodiment, the cutting blade may be heated by a heating device, and then the cutting blade is subsequently moved to cross and thereby cut the tube. In other embodiments, a heating device (such as an RF heater) may directly heat the tube before the cutting blade is moved to cross and thereby cut the tube. Alternatively, the means for removing the section of the tube is configured to remove the section of the tube without directly contacting the tube.

[0062] The automated system may further comprise means for manipulating the tube such that when the tube is removed from the clamping means, the pinched portion formed in the tube remains fluid-tight.

[0063] The automated system may further comprise means for manipulating the tube to release the pinched portion when joined with another such tube, thereby establishing a fluid pathway through the joined tube. In this manner, fluids may be transferred between containers without the need for an operator to release the pinched portion. The means for manipulating the tube to release the pinched portion may be part of any of the robotic devices and / or end effectors described above, or may be a separate component of the automated system.

[0064] The means for manipulating a portion of the tube may further comprise means for sterilizing a second end of the tube.The tube may further comprise an internal valve configured to inhibit (preferably prevent) the flow of fluid or cellular material into or out of the first container through the tube when not connected to another such tube.

[0065] The automated system may further include means for joining the second end of the tube with another such tube. The means for joining the tubes may include means for welding the tubes together to form a tube weld. Welding the tubes together to form a tube weld allows for multiple connections and disconnections to be made while maintaining a closed system. Sterile connections may be made without the need for a sterilant.

[0066] The means for joining the tubes may comprise a connection piece configured to connect between the second end of the tube and another such tube, preferably the connection piece being configured to receive a sterile fluid, e.g. steam, when the tubes are fluidly connected, thereby creating a sterile connection.

[0067] The end effector may include at least one gripping unit configured to engage and move the tube. The tube may include a holding device disposed about the tube whereby the gripping unit grips the holder to engage and move the tube. When the gripping unit grips the holding device, the holding device may be movable along the length of the tube such that the tube may translate (e.g., rotationally or linearly) through the holding device. The tube may have one or more protrusions on its outer surface for engagement by the gripping unit.

[0068] Also disclosed herein is a method of performing a bioprocess in a system having a series of processing stations that perform operations for a bioprocess using one or more containers (such as, for example, the bioprocessing systems described above), the method including configuring an automated system to manipulate a fluid connection between a first container and a separable second container, thereby creating a sterile connection that allows for controlled transfer of fluid or cellular material between the first container and the second container, where manipulating the fluid connection creates a sterile connection that can be separated after transfer of fluid or cellular material is completed, to be able to manipulate a further such fluid connection between the first container and a separable third container, and controlling the automated series of operations of the processing stations.

[0069] Advantageously, by configuring the automation system to make and break sterile connections between the vessels, it is possible to perform a sequence of bioprocessing operations without the need for a human operator. This eliminates human error and allows the automation system to perform the bioprocessing method with great reliability. Additionally, the sterile connections may ensure the transfer of materials between the vessels without exposing the contents of the vessels to the ambient environment. This means that it is not strictly necessary to provide the automation system and the processing stations in a sterile enclosure. The vessels may be relatively simple compared to existing vessels for bioprocessing, since the bioprocessing system may make and break sterile connections. Furthermore, the bioprocessing system may be very flexible, since it may be adapted to process a wide variety of bioprocessing methods (i.e. different sequences of bioprocessing operations). The bioprocessing system is also very scalable, since it is possible to add further processing stations over time.

[0070] The method may further comprise controlling an automated sequence of operations according to a predefined workflow, preferably a reconfigurable bioprocessing workflow. In this way, a specific bioprocessing method can be performed by the bioprocessing system, and the method may be easily modified or adapted without requiring any modifications to the bioprocessing system itself.

[0071] Also described herein is a robotic end effector for joining a first tube to another such tube (preferably via tube welding) thereby forming a fluid pathway, the robotic end effector comprising (e.g., one or more of the above) means for engaging the tube and moving the tube to one or more positions for manipulation, and / or means for clamping a portion of the tube thereby forming a pinched portion of the tube towards an end of the tube whereby the tube is fluidly sealed upstream of the pinched portion, and / or means for removing a section of the tube downstream of the pinched portion thereby removing said end of the tube whereby a new end of the tube not previously in contact with another such tube is formed within the pinched portion, and / or means for joining the pinched portion at the new end of the tube with a corresponding pinched portion of another such tube, and / or means for manipulating the tube to release the pinched portion once joined with another such tube, thereby establishing a fluid pathway between the joined tubes. In one aspect, the robotic end effector may comprise all of these recited features.

[0072] Also described herein is a robotic device for a bioprocessing system (e.g., a bioprocessing system having a series of processing stations that perform operations for a bioprocess using a plurality of separable containers), the robotic device comprising: a base unit configured for automated movement around the bioprocessing system; at least one robotic arm attached to the base unit; and at least one end effector attached to the robotic arm, the at least one end effector configured to perform at least one of: (i) manipulate a fluid connection between two containers, thereby forming a sterile connection enabling controlled transfer of fluid or cellular material therebetween; (ii) manipulate the fluid connection formed between the two containers, thereby performing controlled transfer of fluid or cellular material therebetween; and (iii) separate the fluid connection formed between the two containers after transfer of fluid or cellular material is completed, thereby aseptically sealing each container and enabling formation of further such fluid connections between each container and a different separable container.

[0073] A robotic device (e.g., sometimes referred to as a "mobile manipulation unit") may include an end effector for manipulating fluid connections between tubes (which may include both making and breaking sterile connections) and a separate end effector for transferring (e.g., pumping) fluid along the tubes between containers. Each end effector may be located on a separate robotic arm, or multiple end effectors may be located on the same robotic arm. Alternatively, the robotic device may include a single end effector for both manipulating fluid connections between tubes and transferring fluid along the tubes between containers. The robotic device may further include an end effector for sealing the tubes, such as an RF tube sealing device. Such an end effector for sealing the tubes may be located on a separate robotic arm or may be located on the same robotic arm as one or more of the other end effectors.

[0074] The robotic device may be configured to automatically move around (or within) the bioprocessing system, for example, between processing stations of the bioprocessing system. Alternatively or additionally, the robotic device may be configured for automated movement across a factory floor on which the bioprocessing system is located. The robotic device may be configured to take samples from receptacles of the bioprocessing system, transfer fluids from the receptacles to sampling receptacles, and transport the sampling receptacles to a quality control area (e.g., a quality control (QC) lab). The QC lab may be part of the bioprocessing system or an external QC lab. The robotic device may further comprise a storage area that may be used to store the sampling receptacles within or on the robotic device during transport to the quality control area.

[0075] As used herein, the term "bioprocessing" preferably includes cell therapies, such as autologous and allogeneic cell therapies, as well as vaccines and (small batch) bioprocessing.

[0076] As used herein, the term "automated system" preferably includes a system operated and / or controlled by automation, which preferably includes one or more of a robotic device, a conveyor, one or more actuators configured to engage and / or move containers, or indeed any combination of these features that may move and / or manipulate containers and / or tubes within the system.

[0077] As used herein, the term "robotic device" (or "robot") preferably includes an automated machine or device that is programmed to perform a specific mechanical function, and the term preferably includes a robot, a cobot, an xy robot, a robotic arm, and one or more actuators, possibly including one or more robotic end effectors, and typically including one or more sensors, a microprocessor, and a power source. A robotic device may be located at a fixed location within a bioprocessing system or may be configured to move through several locations within a bioprocessing system. For example, the robotic device may be mounted on rails or may include wheels and / or motors that enable the robotic device to move or drive around the floor of a bioprocessing system, and such a robotic device may be referred to as a "mobile manipulation unit" as described herein.

[0078] As used herein, the term "sterile connection" preferably includes a connection in which the contents of each connected container are not exposed to the surrounding air or atmosphere. The term "sterile connection" may be equivalently referred to, for example, as a "closed connection" or a "sterile connection."

[0079] As used herein, the term "fluid" preferably includes liquids and / or gases, and may further include materials such as cellular material contained therein.

[0080] As used herein, the term "tube" or "tubing" preferably includes a flexible tube or at least one tube having a flexible portion, which may be formed from a thermoplastic or other (e.g., elastomeric) material such as, for example, CFlex®.

[0081] It will be appreciated by those skilled in the art that any apparatus feature described herein may be provided as a method feature, and vice versa. It will also be appreciated that any particular feature, or one or more combinations of features, described and defined in any embodiment described herein may be implemented and / or provided and / or used independently.

[0082] It will be further understood that the invention has been described herein by way of example only, and that modifications in detail may be made within the scope of the invention. Furthermore, as used herein, any "means-plus-function" features may be alternatively expressed in terms of their corresponding structures.

[0083] Exemplary embodiments of the present invention will now be described with reference to the accompanying drawings. [Brief description of the drawings]

[0084] [Figure 1] FIG. 1 is a schematic diagram of a typical bioprocessing workflow. [Figure 2A] FIG. 1 is a schematic diagram of one embodiment of an automated bioprocessing system. [Figure 2B] FIG. 1 illustrates a specific example of a bioprocessing system configured as an automated factory. [Figure 2C] FIG. 1 illustrates a specific example of a bioprocessing system configured as an automated factory. [Figure 3A] FIG. 3 illustrates an example of a mobile robotic device having two robotic arms that may be part of the bioprocessing system shown in FIG. [Figure 3B] 1A-1D illustrate various configurations of a robotic arm and its end effector. [Figure 3C] 1A-1D illustrate various configurations of a robotic arm and its end effector. [Figure 3D] 1A-1D illustrate various configurations of a robotic arm and its end effector. [Figure 4A]FIG. 1 illustrates a tube reel that can provide additional flexible tubing to connect vessels in a bioprocessing system. [Figure 4B] FIG. 1 illustrates an example of a peristaltic pumping device for pumping fluid along a tube. [Figure 5A] FIG. 1 is a cross-sectional view of an example of a flexible tube suitable for use in an automated bioprocessing system. [Figure 5B] FIG. 1 illustrates another example of a portion of flexible tubing suitable for use in an automated bioprocessing system. [Figure 5C] FIG. 1 shows a schematic example of a gripping unit and a portion of flexible tubing adapted to be held thereby. [Figure 6A] 1A-1C show examples of consumables and tubes with identification markings for ease of tracking by a monitoring system. [Figure 6B] FIG. 13 illustrates a tube clip for fastening a tube, the tube clip including an identification mark for facilitating location of the tube by a robotic device. [Figure 7A] FIG. 1 illustrates a consumable suitable for holding media or reagents for use in an automated bioprocessing system. [Figure 7B] FIG. 1 illustrates consumables suitable for use in a centrifuge in an automated bioprocessing system. [Figure 7C] FIG. 1 illustrates a consumable suitable for use as a cell growth vessel in an automated bioprocessing system. [Figure 8A] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8B] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8C] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8D] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8E] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8F] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8G] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8H] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8I] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8J] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8K] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 8L] 1A-1D illustrate a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 9A] FIG. 1 illustrates a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 9B]FIG. 1 illustrates a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 9C] FIG. 1 illustrates a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 9D] FIG. 1 illustrates a first embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 10A] 1A-1D illustrate a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 10B] 1A-1D illustrate a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 10C] 1A-1D illustrate a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 10D] 1A-1D illustrate a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 11A] FIG. 1 illustrates a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 11B] FIG. 1 illustrates a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 11C] FIG. 1 illustrates a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 11D]FIG. 1 illustrates a second embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along a separation process. [Figure 12A] FIG. 1 shows two tubes, each with a flange at the end. [Figure 12B] FIG. 12B shows the tubes of FIG. 12A after flanges have been used to connect the tubes. [Figure 13A] FIG. 12C shows an apparatus for forming the flanges of FIGS. 12A and 12B. [Figure 13B] FIG. 12C shows an apparatus for forming the flanges of FIGS. 12A and 12B. [Figure 13C] FIG. 12C shows an apparatus for forming the flanges of FIGS. 12A and 12B. [Figure 13D] FIG. 12C shows an apparatus for forming the flanges of FIGS. 12A and 12B. [Figure 14] FIG. 13 illustrates a third embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 15A] FIG. 13 illustrates a fourth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at two steps along the connection process. [Figure 15B] FIG. 13 illustrates a fourth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at two steps along the connection process. [Figure 16A] FIG. 13 illustrates a fifth embodiment of an apparatus for forming a sterile connection between tubes having flanges in an automated bioprocessing system at two steps along the connection process. [Figure 16B] FIG. 13 illustrates a fifth embodiment of an apparatus for forming a sterile connection between tubes having flanges in an automated bioprocessing system at two steps along the connection process. [Figure 17A] 13A-13D illustrate a sixth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 17B] 13A-13D illustrate a sixth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 17C] 13A-13D illustrate a sixth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 17D] 13A-13D illustrate a sixth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system at various steps along the connection process. [Figure 17E] FIG. 13 illustrates a sixth embodiment of an apparatus for forming a sterile connection between tubes in an automated bioprocessing system at two steps along a separation process. [Figure 17F] FIG. 13 illustrates a sixth embodiment of an apparatus for forming a sterile connection between tubes in an automated bioprocessing system at two steps along a separation process. [Figure 18A] FIG. 13 illustrates a seventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 18B] FIG. 18B shows a connector used to make the connection of FIG. 18A. [Figure 19] FIG. 13 illustrates an eighth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 20] FIG. 9 illustrates a ninth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 21] FIG. 10 illustrates a tenth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 22A] FIG. 11 illustrates an eleventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 22B] FIG. 11 illustrates an eleventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 22C]FIG. 11 illustrates an eleventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 22D] FIG. 11 illustrates an eleventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 22E] FIG. 11 illustrates an eleventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 22F] FIG. 11 illustrates an eleventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 22G] FIG. 11 illustrates an eleventh embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23A] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23B] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23C] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23D] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23E] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23F] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23G] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 23H] FIG. 12 illustrates a twelfth embodiment of an apparatus for forming sterile connections between tubes in an automated bioprocessing system. [Figure 24A]FIG. 1 illustrates one embodiment of an apparatus for forming a sterile connection between consumables in an automated bioprocessing system. [Figure 24B] FIG. 1 illustrates one embodiment of an apparatus for forming a sterile connection between consumables in an automated bioprocessing system. [Figure 24C] FIG. 1 illustrates one embodiment of an apparatus for forming a sterile connection between consumables in an automated bioprocessing system. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0085] An example of a typical bioprocessing workflow is shown in FIG. 1. The process includes a number of vessels and reagents, with each arrow indicating a fluid transfer between vessels. Due to the concerns already mentioned above, conventional bioprocessing systems provide all of the vessels and fluid paths within a single consumable. While this can mitigate concerns about cross-contamination between vessels, complex consumables are extremely expensive and provide no flexibility in modifying the bioprocessing workflow. Due to the large number of integrated valves and pumps, the consumables can also be unreliable. Therefore, by splitting the workflow into much smaller parts (such as those shown by the dotted lines), as will be described in detail here, it is possible to achieve the same result with a much less complex consumable. This also allows for much more flexibility when the workflow is modified and allows for the extraction of samples throughout the process for quality control.

[0086] An exemplary embodiment of a bioprocessing system 1 according to the present invention is shown in Figure 2A. The bioprocessing system 1 comprises a series (e.g., "plurality") of processing stations 20 configured to perform processing steps for a bioprocess, and an (automation) system 1a for (at least partially) automating the process.

[0087] In this exemplary embodiment, the bioprocessing system 1 has processing stations 20 in the form of a thawing station 4, a centrifuge 6, a magnetic cell separator 8, a controller rate freezer 10, and a refrigerator 11, although additional alternative stations 20 for processing (not shown) may be provided depending on the particular process being performed by the bioprocessing system 1. Also, there may be multiple instances of any given processing station 20 at separate locations within the bioprocessing system 1.

[0088] The processing stations 20 may include any combination of concentration stations, cryopreservation units, washing stations, cell concentration stations, cell growth stations, cell selection stations, stations for determining cell count, cell viability, cell phenotype or cell type, e.g., cytometer stations, and / or stations for any other suitable processing or analysis steps. The bioprocessing system 1 also has an incubator 12 large enough to accommodate and incubate multiple consumables 13 at a time, including under perfusion. Advantageously, the cytometer station may facilitate automatically taking samples to obtain intermittent readings of cell count and quality. This is beneficial for keeping the process under control and initiating process improvements through adaptive control, potentially allowing for further predictive elements, as will be described in more detail below.

[0089] For example, the incubator 12 may store 20 consumables 13 and operate at approximately 37° C., although the number of consumables 13 may be selected to meet the needs of the particular bioprocess being performed. Additional incubators 12 may be provided at separate locations within the bioprocessing system 1 to provide additional space for additional consumables 13. Each consumable 13 may contain a cell sample, a reagent, or a fluid, and each consumable 13 connects to a first end of a tube 150 that leads to a second end of a tube (150 not shown) and is fluid-sealed when unconnected (or “free”). Thus, as referred to herein, a “consumable” may be in the form of a “container” that may hold, for example, cellular material to be processed in a cell therapy process.

[0090] All of the consumables 13 and reagents may be pre-loaded into the bioprocessing system 1 before a particular bioprocess is initiated, although additional reagents may be added throughout the process as needed (e.g., on day 7 of a 10 day therapy process). Additional reagents may be needed, for example, for reactivation of cells or to add additional media to the consumables 13.

[0091] A particular bioprocess is defined by a bioprocess workflow, and preferably, the bioprocess system 1 may be configured to perform several bioprocess workflows. For example, the bioprocess system 1 may perform the same bioprocess workflow on multiple patient samples in parallel, or perform different bioprocess workflows on multiple patient samples in parallel. Each bioprocess workflow may use a different subset of the processing stations 20 in the bioprocess system 1. In a preferred embodiment, the bioprocess system 1 comprises stations 20 for performing concentration, washing and incubation processes.

[0092] The bioprocessing system 1 comprises an automation system 1a configured to place one or more consumables 13 at each of a series of processing stations 20 and move the consumables 13 between the stations 20. In this embodiment, the automation system 1a includes at least one robotic device 2 that can move the consumables 13 between the various stations 20 and manipulate the tubes 150 connecting each of the consumables 13. Alternatively or additionally, the robotic device 2 may be configured to move the processing stations 20 to connect the consumables 13 to the processing stations 20. The bioprocessing system 1 may further comprise a viewing system 35 (e.g., a machine vision system) for viewing the operation of the automation system 1a. The bioprocessing system 1 may further comprise a processing and control unit 38 for controlling the sequence of operations of the automation system 1a, in other words the processing and control unit 38 may provide a means for controlling the automated sequence of operations of the automation system 1a including the processing stations 20 and / or the robotic device 2. Although the processing and control unit 38 is described herein as an individual unit of the bioprocessing system 1, it will be understood that there may be multiple units, e.g., separate units for processing and for control.

[0093] A specific example of a bioprocessing system 1 is shown in Figures 2B and 2C, where all the processing stations 20 described above are located in a static position on the factory floor 17. In this configuration, the bioprocessing system 1 may be referred to as an automated factory. Each robotic device 2 is also provided on the factory floor 17, which may be implemented in several ways. For example, each robotic device 2 may be located in a fixed position in the bioprocessing system 1 or may be mounted on rails 18, which allows the robotic device 2 to access all areas of the bioprocessing system 1, such as the stations 20. The robotic device 2 may be configured as a collaborative robot ("cobot"). The robotic device 2 may have at least one robotic arm 3 for manipulating the consumables 13 and the tubes 150, as shown herein, or may include a conveyor belt, one or more actuators, or any combination of the above aspects. Preferably, the bioprocessing system 1 comprises multiple robotic devices 2, and different processes of the bioprocessing system 1 are performed by different robotic devices 2. For example, there may be separate robotic devices that manipulate the tubing 150, form fluid connections between separate consumables 13 (e.g., by tube welding and sealing), pump fluids along the tubing 150, transport the consumables 13, and / or sample from the consumables 13 for quality control. It will be understood that other operations may be performed by the robotic devices 2, and that any of the robotic devices 2 may be configured to perform more than one operation. Preferably, multiple robotic devices 2 may be configured to perform each operation to provide a degree of redundancy, allowing any robotic device 2 to be repaired and / or replaced without interrupting the progression of a particular bioprocess workflow.

[0094] Each robotic device 2 may be implemented as a mobile operational unit 2 as shown in FIG. 3A. The mobile operational unit 2 may be configured to move autonomously across the floor 17 of the bioprocessing system 1 to access areas such as the stations 20. Each mobile operational unit 2 may include a base unit 2a configured for automated movement within or around the bioprocessing system 1. For example, the mobile operational unit 2 may include a wheel-mounted base unit 2a that may house a motor and other control and communication components (not shown) that are configured together to enable the mobile operational unit 2 to move within or around the floor 17 of the bioprocessing system 1. For example, the mobile operational unit 2 may include a communication unit (not shown) that may receive instructions from and / or transmit data (such as from a sensor or camera) to the processing and control unit 38, which acts to control the motor to drive the wheels, thereby driving the mobile operational unit 2. The use of the mobile operational units 2, particularly for moving consumables around the factory floor 17, is advantageous because it allows asynchronous processes to be performed simultaneously, unlike conveyors that require precise sequences of motion.

[0095] Each mobile operational unit 2 may have at least one robot arm 3 attached to the base unit 2a and configured to perform one or more operations in the bioprocessing system 1. Preferably, the mobile operational unit 2 may have multiple robot arms 3 attached to the base unit 2a (e.g., two robot arms 3 as shown in the example of FIG. 3A), each configured to perform different operations, such that multiple operations may be performed by the mobile operational unit 2 when placed in the processing station 20, e.g., sequentially or simultaneously. For example, any robot arm 3 may be configured to operate a fluid connection between consumables, and another robot arm 3 may be configured to pump fluid along a tube 150. There may also be a robot arm 3 that seals the tube 150. Each robot arm 3 may operate at least one end effector 100 configured to perform at least one operation in the bioprocessing system 1.

[0096] Various exemplary configurations of the robot arm 3 will now be described in relation to Figures 3B-3D. These configurations may be incorporated into a mobile robotic device 2 (e.g., the mobile operational unit 2 described above) or into a stationary robotic device 2 located at a fixed position within the bioprocessing system 1. In the embodiment shown in Figure 3B, the robotic device 2 comprises a first robotic arm 3-1 having a first end effector 100-1 and a second robotic arm 3-2 having a second end effector 100-2. This allows the mobile operational unit 2 to be flexible and both robotic arms 3-1, 3-2 can be operated independently. In this example, the first end effector 100-1 is configured as a sterile tube welder in a manner described in detail below, and the second end effector 100-2 is configured as a pumping unit 30 for pumping fluid through a tube 150. One embodiment of the pumping unit 30 will be further described in relation to Figure 4B. However, when the robot device 2 is a stationary robot device 2, it is difficult for both arms 3-1, 3-2 to reach all required areas.

[0097] In an alternative embodiment, as shown in Figure 3C, a single robotic arm 3 includes an end effector 100 having a first portion 100a configured as a sterile tube welder and a second portion 100b configured as a pumping unit 30. In this manner, the single robotic arm 3 can simply change position (e.g., rotate) so that each portion 100a, 100b can be operated for its specific purpose. This improves the reach of the robotic arm 3 and increases space efficiency, but may result in a heavier end effector 100.

[0098] As a further alternative, the end effector 100 may be interchangeable with the robotic arm 3. As shown in FIG. 3D, the robotic arm 3 may have a gripping unit 50 configured to engage with a selection of end effectors 100 as already described above. The gripping unit 50 may have a pair of jaws for gripping the selection of end effectors 100 or may use a magnetic coupling to select one of the end effectors 100. The selection of end effectors 100 may include an end effector 100-1 configured as a tube welder, an end effector 100-2 configured as a pumping unit 30, an end effector 100-3 configured as a sealing unit, or any combination of end effectors. The selection of end effectors 100 may be located anywhere in the bioprocessing system 1 accessible by the robotic device 2. Preferably, the selection of end effectors 100 is always located within the reach of the robotic arm 3, such as in a tool holder 3a next to the robotic arm 3 or in a "tool belt" 3a. Advantageously, this allows a single robotic arm 3 to operate a wide range of specialized end effectors 100.

[0099] For example, the mobile operating unit 2 may have end effectors 100 for both tube welding and pumping. Thus, the mobile operating unit 2 may be able to join two consumables 13 together and transfer fluid between the consumables 13. Alternatively, or in addition, the mobile operating unit 2 may have end effectors 100 for tube sealing. Alternatively, or in addition, the mobile operating unit 2 may take a sterile sample (e.g., from a bioreactor) by welding and pumping fluid into the sampling consumable 13. The mobile operating unit 2 may then transport the sampling consumable to a quality control (QC) lab. The mobile operating unit 2 may include a storage area (not shown) in which the sampling consumable 13 may be stored during transport to the QC lab. Preferably, the storage area is temperature controlled.

[0100] The automated system 1a is configured to manipulate a fluid connection between a first consumable 13 and a separable second consumable, thereby creating a sterile connection that allows for the controlled transfer of fluid or cellular material between the first consumable 13 and the second consumable 13. Here, the robotic device 2 is used to form (or manipulate) the fluid connection between tubing 150 so that the separate consumables 13 may be connected together.

[0101] The connection between the tubes 150 can be performed by the end effector 100 arranged on the robot arm 3, as already mentioned above. Alternatively, the robot arm 3 may move and place the tube in a separate connection unit (not shown) of one of the stations 20 to which the tube 150 is subsequently connected. In either case, the connection between the tubes 150 is performed aseptically, whereby the consumable 13 and the contents of the tube 150 are not opened or exposed to the surrounding air or atmosphere at any stage, i.e. the connection remains "closed" and does not require additional sterilizing agents to prevent contamination. Several ways of forming or maintaining a "closed" sterile connection between the consumables 13 are described in detail below. However, preferably, sterile tube welding is used to manipulate the fluid connection between the tubes 150.

[0102] The fluid connections are also reversible, such that the tubing 150 can be disconnected and reconnected to different consumables 13 as many times as necessary to perform the required bioprocessing method. In other words, the automated system 1a is configured to create a sterile connection that can be disconnected after the transfer of fluid or cellular material is completed, so as to be able to operate further such fluid connections between a first consumable and a third consumable that can be disconnected.

[0103] As described above, during both connection and disconnection, the consumables 13 and tubing 150 do not have their contents exposed to the surrounding air or atmosphere, thereby allowing controlled transfer of fluid and / or cellular material to occur only between the consumables 13 that are connected together.

[0104] It is desirable to keep the length of the tubing 150 connecting to each consumable 13 relatively short to avoid entanglement between the different tubes 150 and other parts of the bioprocessing system 1. However, it is also important to have a sufficient supply of tubing to make many connections and separations between several separate consumables throughout a particular cell therapy method. Therefore, a tube supply means 19, such as a tube reel 19, may be provided in the bioprocessing system 1. An example of a tube reel 19 is shown in FIG. 4A. The tube supply means 19 may be provided in a fixed position within the bioprocessing system 1 or may be integrated into at least one of the robotic devices 2, such as directly on the end effector 100. If the tube 150 connecting to a particular consumable 13 is not long enough to make the necessary connections in a particular bioprocessing step, the automation system 1a may extract additional lengths of tubing from the tube supply means 19 and use it to extend the tube 150 connecting to the consumable 13. As will be described in more detail below, the extension of tube 150 is preferably performed by tube welding in a manner that ensures that the contents of tube 150 are not exposed to either the environment or the remaining length of tubing within tube supply means 19.

[0105] Since the contents of the tubes 150 are never exposed to the environment, there is no need to have a sterile atmosphere around the stations 20, consumables 13, or robotic devices 2. An enclosure 14 may be provided to prevent access by an operator and / or to provide a sterile atmosphere or to control the environment, for example by controlling temperature, light levels or other conditions. However, preferably, the bioprocessing system 1 does not require a sterile enclosure 14, and instead the processing stations 20 are provided on the factory floor 17 in a space that can be traversed and accessed by both human operators and one or more robotic devices 2.

[0106] The bioprocessing system 1 also comprises a pumping unit 30, which pumps the fluid along the tube 150 once the robotic device 2 has successfully connected the two consumables 13 via the respective tubes 150. As already mentioned above, the pumping unit 30 may be located on the robotic arm 3. Alternatively, the pumping unit 30 may be a stationary component located on one of the stations 20 where the tube 150 is placed by the robotic arm 3 for pumping to take place. The pumping unit 30 may be a peristaltic pumping device 30 as shown in FIG. 4B, but alternatively or additionally a pressure-driven flow pump or a syringe pump may be used. Optionally, different types of pumps may be used for the different pumping operations. This pumping device 30 comprises a rotating wheel 31 driven by a motor on a shaft (not shown). The pumping device 30 also comprises a clamp 32, and prior to the pumping operation the tube 150 is positioned between the rotating wheel 31 and the clamp 32 by the robotic device 2. The pumping device 30 then compresses the tube 150 between the rotating wheel 31 and the clamp 32, pumping the fluid along the tube 150 as the rotating wheel 31 rotates. Additionally, the pumping device 30 may be used to prevent fluid flow through the tube 150 by compressing the tube 150 between the rotating wheel 31 and the clamp 32 without rotating the wheel 31. While a pumping unit 30 is preferred, the transfer of fluid and cellular material may also be accomplished by gravity, for example, or by the addition of gas through a sterile filter.

[0107] The robot arm 3 may have at least one gripping unit (50, not shown) that allows the robotic device 2 to hold and move the consumables 13 and the tubes 150. The tubes 150 are made sufficiently flexible so that they can be manipulated into position to be welded. The tubes 150 preferably have a standardized material, shape and diameter so that connections between the tubes 150 can be performed reliably by the robotic device 2. For example, the bioprocessing system 1 may use only one standardized type of tube throughout the system, or may use a small number of standardized tubes. The types of tubes may be selected to optimize the weld quality. Reliability may be improved by using a small number of pre-specified tubes that have been verified to weld extremely well.

[0108] 5A shows a cross-section of tube 150 having a non-circular profile, such that tube 150 may be easily manufactured by attaching two flat strips of material together. Furthermore, tube 150 may be easily flattened by gripping unit 50 or by pumping unit 30 to pinch tube 150 closed to pump fluid through tube 150 or to prevent fluid migration. Tube 150 is preferably formed from a thermoplastic resin, but may be formed from other materials, such as a CFLex® (elastomer) material.

[0109] Each tube 150 may have a section surrounded by a rigid outer casework that can be easily manipulated by the robotic arm 3. Alternatively, the tube 150 may have a series of projections spaced along its exterior length that are easily manipulated. For example, Figure 5B shows a tube 150 having a series of projections 40 (e.g., "handling sections") in the form of radially (outwardly) extending flange regions 40 preformed at various locations along its length.

[0110] Providing the handling section 40 may facilitate applying tension to the tube 150. For example, Figure 5C shows a gripping unit 50 having two grippers 55 that may be used to grip the handling section 40. In this manner, the grippers 55 may be disposed on the pumping unit 30 or on another piece of equipment, such as a pinch valve (not shown) to prevent fluid flow, to apply tension to the tube 150 and straighten it.

[0111] The bioprocessing system 1 may also include image capture systems or devices, such as sensors and / or cameras, that are used during operation of the bioprocessing system 1 or for inspection and quality control. Together with one or more processing units, these image capture systems or devices may be referred to as observation systems or machine vision systems 35, while processing may be performed by a processing and control unit 38. For example, the cameras may be distributed throughout the bioprocessing system 1, such as in fixed locations on the factory floor 17. Alternatively, or in addition, the cameras and sensors may be located on one or more robotic devices 2, such as on at least one robotic arm 3 or end effector 100 of the robotic device 2.

[0112] As shown in FIG. 6A, each consumable 13 can be identified by a camera using an identification mark 13-1, such as using a unique barcode or QR code on each consumable 13. This allows all consumables 13 and samples to be uniquely and automatically tracked throughout the cell therapy process, maintaining traceability and facilitating integration with electronic batch records (EBRs) and allowing processing of multiple batches at the same time. Other identification methods can be used, such as using unique radio frequency identification (RFID) tags. It is important to be able to accurately and automatically track samples throughout the bioprocessing system 1, especially when the bioprocessing system 1 processes several patient samples, to reduce the risk of mixing different batches.

[0113] Similarly, the tube 150 may also be identified using an identification mark 150-1. For example, the identification mark 150-1 may be a unique barcode or QR code, but other identification methods may be used, such as using an RFID tag. The identification marks 150-1 may be placed at regular intervals along the length of the tube 150, with each identification mark 150-1 uniquely providing data about the tube 150, such as its location, orientation, material, size and / or other characteristics. For example, the identification mark 150-1 may indicate a unique reagent ID, the size and material of the tube (which may affect welding and pumping parameters), the distance between the identification mark 150-1 and the corresponding container 13, and / or the orientation of the tube (such as to indicate which direction is toward the container 13). The identification mark 150-1 may be printed on or laser etched into the tube 150 in a manner similar to how manufacturer data is marked. Providing the identification marks 150-1 along the tube 150 may provide several advantages. First, it is possible to reliably identify which particular tube 150 is connected to which vessel 13 simply by inspecting the identification mark 150-1 on the tube. This ensures that before a sterile connection is made between two vessels 13, the tubes 150 are first brought together and checked by one of the cameras in the bioprocessing system 1 before the connection process begins, thereby reducing the risk of the vessels 13 being connected incorrectly. Furthermore, it is also possible to determine where along the length of the tube 150 the connection should be made, thus minimizing tube waste and potentially calculating the number of cuts based on the total remaining length of the tube 150. Finally, the identification mark 150-1 may also be used to indicate to the processing and control unit 38 the settings to be used for welding and pumping.

[0114] FIG. 6B shows a tube clip 41 clipping a tube 150. Several such tube clips 41 may be distributed throughout the bioprocessing system 1. The tube clip 41 may include an identification mark 41-1 that uniquely identifies the location of the tube clip 41 in the bioprocessing system 1. For example, the identification mark 41-1 may be a unique barcode or QR code, but other identification methods may be used, such as using a unique RFID tag. The tube clip 41 includes a tube holding element, such as at least a pair of clip jaws 41-2, that holds the tube 150 in place when the tube is inserted between the clip jaws 41-2. The clip jaws 41-2 are positioned in a fixed position relative to the identification mark 41-1 so that the tube 150 held by the clip jaws 41-2 can be reliably positioned with reference to the identification mark 41-1 alone. Advantageously, this simplifies the task of autonomously positioning and handling the flexible tube 150, and instead allows the gripping unit 50 of the robot arm 3 or end effector 100 to move to a precisely defined set of coordinates (e.g., XYZ coordinates) to position the tube 150 between the clipping jaws 41-2. The tube clip 41 may further comprise sensors, such as air bubble sensors, that may detect the presence of a tube, air in the tube, and / or the flow rate of fluid through the tube 150. Furthermore, the tube clip may also include actuators for moving the clip from open and closed positions, the movements between which are designed to seat the tube at a defined reference point and prevent the tubing from being pulled out of the clip. The tube clip 41 may be located at a fixed position (e.g., a predetermined, known, or easily identifiable position) within the bioprocessing system 1. The tube clip 41 may be movable before / after / during a bioprocessing operation. For example, the tube clips 41 may be initially attached to the bioprocessing system 1 by a user during the setup phase or may be attached to a rail or rack, and then the user may clip one or more tubes 150 to the corresponding tube clips 41.The machine vision system 35 can scan the tube clips 41 and identify the initial positions of all tubes 150 so that the robotic device 2 can properly engage the tubes 150 during bioprocessing operations. This results in a flexible and robust setup.

[0115] For quality control, the camera and sensors of the machine vision system 35 may inspect the connections between the tubes 150 to verify that the connections have been successfully made. The camera may have a microscope lens that allows detailed inspection of the connections between the tubes 150. During inspection by the camera and sensors, the connections may be tested in several ways. Ultrasound may be used to check if there are any voids in the connection and / or the gripping unit 50 may be used to apply pressure to the tubes 150 at or near the connection. The gripping unit 50 may be used to apply tension to the connection between the tubes 150 and measure the stress-strain profile of the joined tubes 150. Fluid sensors or air sniff sensors (e.g., "sniff leak" or "gas leak" detectors) may be used to detect fluid leakage from the connection. If the measured stress-strain profile, visual inspection by the camera, or parameters measured by the sensors indicate that the connection between the tubes 150 is poor, the tubes 150 may be separated and a new fluid connection may be manipulated until a sterile connection is successfully formed. Quality control may be performed automatically, without input from an operator, each time a connection is made. The connections between the tubes 150 may be isolated from their respective consumables 13 until a quality control is performed. This may be accomplished by pinching the tubes 150 and / or by only allowing fluid to flow out. In this way, even if a bad connection is discovered, the contents of the consumables 13 still remain isolated from the surrounding air or atmosphere. In the event of a bad connection, the process may be repeated until a good connection is made before process material enters the connection area.

[0116] The bioprocessing system 1 may further comprise a processing and control unit 38, which may be configured to execute one or more software programs and / or to control various components of the bioprocessing system 1, such as the automation system 1a, the processing station 20, and / or the machine vision system 35. Although the processing and control unit 38 is described herein as a single unit, it will be understood that multiple units may be provided to perform the same function, such as separate units for processing and for control. The bioprocessing system 1 may have a user interface 15 through which a user inputs instructions to be executed by the processing and control unit 38. The user interface 15 may also be located remotely to allow remote monitoring and / or control of the bioprocessing system 1, for example with data stored in the "cloud". The bioprocessing system 1 has a loading hatch 16, through which new consumables 13 may be loaded into the bioprocessing system 1, or equivalently, used consumables 13 may be removed from the bioprocessing system 1 after use. An operator can also use the user interface 15 to program the bioprocessing system 1 to perform a particular automated sequence of operations in a particular bioprocessing workflow, thereby providing a means to control the automated sequence of operations of the processing stations of the bioprocessing system 1. The operator can also use the user interface 15 to automatically take routine samples from the process, which may be a cell count processing station or process on a cytometer, or may be removed from the bioprocessing system 1 via the loading hatch 16 without exposing any of the contents of the consumables 13 to the environment. Samples may be run on other third party equipment (sometimes referred to as a QC lab), such as to test for cell count, viability, or any other parameter to monitor the progress of the cell therapy process.By analyzing samples throughout the cell therapy process, operators can ensure that the process is maintained within specifications, and further, the resulting data allows for adaptive control, such as adjustment of gases, media, and other parameters for each consumable 13 in the process.

[0117] An additional problem associated with the automated manipulation of the tubes 150 is that the free ends of the tubes 150 are difficult for the automation system 1a to identify and may be in an uncertain position. In particular, when manipulating long lengths of tubing, there is a risk that the tubes 150 may become entangled with each other or collide with other parts of the bioprocessing system 1 as they move around the bioprocessing system 1. Therefore, the processing and control unit 38 may operate the automation system 1a such that the movement of the tubes 150 generally follows a well-defined path between set positions, so that the behavior may be well characterized and confirmed. In other words, the sequence of operations of the bioprocessing workflow may include a list of defined unit operations and connection steps, which are repeatable and reversible by the automation system 1a, such that the tubes 150 are moved from any known position to another known position along a predefined path.

[0118] Furthermore, if the free end of the tube 150 needs to be positioned, the automation system 1a may first position the corresponding consumable 13 and then follow (visually or mechanically) the tube 150 until the free end of the tube 150 is positioned. If this is performed mechanically, such as by one of the robotic devices 2, the robotic device 2 may straighten the tube 150 so that the robotic device 2 still follows the predetermined path. Similarly, when performing a welding operation, it may be desirable to move the tube 150 and connect the tube in a free space, such as a position between two corresponding containers. To enable this, the robotic device 2 may pull the tube 150 through the tube clip 41 (e.g., when the tube clip 41 is in a closed position, as outlined above), so that the robotic device 2 still follows the predetermined path, and further, this allows stresses or strains on the tube to be absorbed by the tube clip 41. In other words, the tube clip 41 may act as a pulley or bracket that may pull or translate the tube 150 .

[0119] The processing and control unit 38 may execute an automatic scheduling program that automatically schedules a series of operations to be performed by the bioprocessing system 1. One challenge associated with performing multiple cell therapy operations in parallel is that each of the operations may start at different times, take different times to execute (due to biological variations), and have different programmed workflows. Additionally, the bioprocessing system 1 will have a limited number of resources, such as processing stations 20, robotic devices 2, and / or robotic arms 3. As a result, many conflicts may arise when scheduling multiple cell therapy operations, and there may be a significant risk of mechanical collisions between parts of the bioprocessing system 1.

[0120] To address this, the auto-scheduling program translates the various user-programmed workflows and determines a course of action for the bioprocessing system 1 to follow. The auto-scheduling program may update this course of action based on inputs such as the current processing time. If the auto-scheduling program determines that two conflicting actions must be performed simultaneously, the program may delay one of the tasks within specified limits to avoid the conflict. If this is not possible, the program may instead delay a less important task or flag an error or raise an alarm for human intervention. The importance of a task may be determined based on a pre-programmed or user-configurable list of priorities.

[0121] Furthermore, the processing and control unit 38 executes a simulation program that may simulate the workflow and the corresponding sequence of actions both before and during a run, determine when future events will occur, and may further determine the likely quality of the output product based on the input material properties. The simulation program may have means to simulate process variations and may have means to update knowledge of future variations based on historical data and user input parameters. Process variations may result from biological variations, human operator variations, and / or machine variations. The simulation program may prevent the start of a run if it predicts that a conflict will occur, and may indicate when a manual step needs to be performed by the operator (e.g., taking a sample to an external QC lab). The user interface 15 may also have means to warn the user about the minimum time they must wait before the next patient run can begin, and to highlight all of the interdependencies of the actions.

[0122] The automation system 1a may move outside of the validated parameters due to human error during set-up, interference with the production line, unexpected part movements, variations in the length of the bioprocess, variations in the arrival of input materials, and / or noise in the production system. Furthermore, the software in the processing and control unit 38 is necessarily complex since it performs many complex functions simultaneously in parallel. This may make it difficult to validate and verify the bioprocessing system 1, especially when certain reliability and safety requirements need to be met. To address this issue, the processing and control unit 38 may also execute a validation program (or a "witness system") to verify whether the bioprocessing system 1 is performing correctly as intended. For example, the validation program may verify that the correct sequence of actions has been performed, but may also compare the actual positions of the robotic device 2, tubing 150, connections, and fluidic equipment with their intended positions. If the validation program observes that the automation system 1a is not performing as intended, it flags an error and / or issues an alarm. The processing and control unit 38 may use the validation program in combination with the simulation program to determine whether the automation system 1a is likely to be out of specification in the future and / or to predict future performance. If the predicted future performance is likely to be out of specification, the processing and control unit 38 may issue an alarm or take action to bring the automation system 1a back within specification. Preferably, the validation program receives data input from a separate set of sensors for use in the machine vision system 35 and is run as a separate process to avoid a single point of failure. In other words, the machine vision system 35 may include a separate subset of cameras and / or sensors that provide input to the validation program.

[0123] By allowing reversible fluid connections between the consumables 13, each consumable 13 can have a simpler structure and be manufactured at a lower cost than previous consumables. Since the automated (robotic) system 1a can perform all steps required to perform a complete cell therapy process without human intervention, human error is eliminated and the automated (robotic) system 1a can perform the steps very reliably. Furthermore, since all consumables 13 can be disconnected and reconnected at any time, multiple cell therapy processes can be performed in parallel. Similarly, an operator can instruct the bioprocessing system 1 to start a new therapeutic process at any time as long as the bioprocessing system 1 is not full. Additionally, since the robotic device 2 can connect any two consumables 13, the process can be easily adapted to introduce additional steps or to perform an entirely different method of cell therapy. To do so, the bioprocessing system 1 may be programmed to include different or additional steps and use additional consumables 13 or stations 20. For example, the bioprocessing system 1 may perform a method of cell therapy, such as CAR-T, NK cell, Treg therapy, HSC or any other suitable process.

[0124] An example of a cell therapy process that may be performed by the bioprocessing system 1 will now be described.

[0125] First, the operator loads a set of consumables 13 through the loading hatch 16. These consumables 13 include a processed blood sample contained in a patient leukoreduction pack (leukopack), bags for media and reagents, and a bag for receiving waste.

[0126] After loading the consumable 13, the operator programs the desired cell therapy process via the user interface 15. First, the robotic device 2 places the leukopack in the thawing station 4 to thaw the contents of the leukopack. Then, the end effector 100 of the robotic device 2 operates a sterile connection between the leukopack and the consumable 13, and the pumping unit 30 transfers the contents of the leukopack to the consumable 13 via the sterile connection. The robotic device 2 moves the consumable 13 to the cell washer 6, which may be a centrifuge such as a drum-based centrifuge 6, a counterflow centrifuge, or a spin membrane type device. The robotic device 2 sequentially makes several connections between the consumable 13, the media bag, and the waste bag to wash the sample multiple times with buffer solutions. For example, the consumable 13 may be washed three times in this manner. The blood sample is then transferred from the consumable 13 to a temporary holding bag so that density gradient medium is added to the consumable 13 from one of the reagent bags before the blood sample is returned to the consumable 13 where density gradient separation is performed.

[0127] Here, the blood sample is transferred to a fresh consumable 13 and a further sterile connection is made by the robotic device 2 to add the activation reagent. The robotic device 2 gently rocks and / or rotates the consumable 13 to mix the activation reagent with the blood sample before transferring the consumable 13 to the incubator 12 for 24 hours. The consumable 13 is then removed from the incubator 12 and the blood sample is transferred to the RetroNectin-containing consumable 13 followed by the addition of the viral vector. This consumable 13 is returned to the incubator 12 for 24 hours. After the robotic device 2 removes the consumable 13 from the incubator 12, the robotic device 2 transfers the blood sample to a consumable 13 suitable for use in the centrifuge 6. After the consumable 13 is removed from the centrifuge 6, the blood sample may be washed again several times by adding buffer from the media bag and removing the waste to the waste bag.

[0128] The blood sample is then transferred to a growth container consumable 13 connected to a perfusion system and placed in an incubator 12 for 7 days for cell growth. Finally, the blood sample is removed from this consumable 13 and transferred to another consumable 13 which allows the blood sample to be concentrated in a centrifuge 6 before being transferred to an infusion bag where cryoprotectant and other formulation additives are added. This infusion bag is then placed in a controlled rate freezer 10 for frozen storage before being returned to the operator through a loading hatch 16.

[0129] Although the exemplary automated process described above follows several steps and requires the use of multiple consumables 13, each of the consumables 13 can be quite simple in its form. For example, a bag for medium and reagents can be like the consumable 300 shown in FIG. 7A, having an inlet / outlet 301 to which the tube 150 is connected. FIG. 7B shows a consumable 310 suitable for use in a centrifuge 6, which not only has an inlet / outlet 311 but also a sterile air filter 312 and a vacuum-operated tap 313 for drawing fluids in the chamber during use of the centrifuge 6. Examples of suitable consumables, centrifuge containers and centrifuges can be found in EP 1 144 026 and US 10 562 041, which are well known and therefore do not need to be described further here. FIG. 7C shows a consumable 320 suitable for use as a growth container for a cell growth process. It has an inlet 321 for medium 326, an outlet 322 for waste and an inlet / outlet 323 for cell inoculation, sampling and / or cell harvesting. The growth vessel 320 contains cells 325 and a gas permeable membrane 324. As previously explained, these consumables 13 can be manufactured much more reliably and at a much lower cost than prior art consumables. Thus, the bioprocessing system 1 can provide an automated cell therapy process with (substantially) no human intervention.

[0130] With reference to Figures 8A-8L, a preferred embodiment of a method for manipulating a fluidic sterile connection between tubes 150 will be described in detail. Here, a robotic device 2 of a bioprocessing system 1 comprises a robotic arm 3 having an end effector 100. The end effector 100 is attached to the robotic arm 3 and has two gripping units 110a, 110b. The gripping units 110a, 110b may be the gripping parts 55 of the gripping unit 50 described above or may be separate gripping units. The end effector 100 is configured to connect a first tube 150a and a second tube 150b together while maintaining a seal between the inside of the tubes 150a, 150b and the surroundings (i.e., the contents of the tubes and consumables are not exposed to the atmosphere). Each tube 150a, 150b connects to a respective consumable 13 (not shown). As used herein, the term "upstream" refers to a direction along the tubes 150a, 150b toward a first end of the tube 150a, 150b that is attached to the respective consumable 13. Similarly, the term "downstream" refers to a direction along the tubes 150a, 150b toward a second "free" end of the tube 150a, 150b. Attached to each tube 150a, 150b is a tube holder 130a, 130b that can be easily gripped by the gripping units 110a, 110b. The tube holders 130a, 130b may be equivalently referred to as "holding devices" or "holders." The tube holders 130a, 130b may move along the tubes 150a, 150b via rotation of the precession wheels 135a, 135b (i.e., translating the tube 150 relative to its respective tube holder 130). The end effector 100 includes a clamping unit 105 having a first jaw 120 and a second jaw 125 divided into a first portion 120a, 125a and a second portion 120b, 125b, respectively. The end effector 100 also includes a blade 140 that can move between portions of the jaws 120, 125 along a cutting surface.

[0131] In FIG. 8B, gripping units 110a, 110b are shown gripping respective tube holders 130a, 130b on respective tubes 150a, 150b, which are positioned adjacent to the clamping unit 105 with the jaws 120, 125 of the clamping unit 105 in the open position.

[0132] In FIG. 8C, the precession wheels 135a, 135b rotate to advance the tubes 150a, 150b through the jaws 120, 125. The camera 160 is used to verify that each tube 150a, 150b is correctly positioned within the clamping unit 105 and that both tubes 150a, 150b intersect the cutting plane. The tube holders 130a, 130b may include magnets to facilitate alignment of the tubes 150a, 150b within the clamping unit 105. The jaws 120, 125 of the clamping unit 105 may be coated with a low friction material so that the tubes 150a, 150b slide within the clamping unit 105 and the effects of Poisson's ratio are minimized.

[0133] In Figure 8D, the jaws 120, 125 of the clamping unit 105 are clamped together to pinch the tubes 150a, 150b flat at the cut surface, thereby preventing fluid flow through the tubes 150a, 150b. Because all tubes 150 in the bioprocessing system 1 are identical in size, the jaws 120, 125 of the clamping unit 105 are configured to be rigid, such that when clamped, they completely encase the tubes 150a, 150b with controlled tolerances to perfectly define the tube form factor and alignment, regardless of tube tolerances. The tubes 150 may have pre-formed flange areas, such as large flat flange areas, to facilitate alignment with the clamping unit 105.

[0134] In Figure 8E, pumping unit 30 is used to pump fluid in the direction of the arrow and away from clamping unit 105. This ensures that both tubes 150a, 150b are completely dry at the cut surface and helps to further fold and pinch tubes 150a, 150b to keep them closed.

[0135] In Figure 8F, blade 140 is heated by a heat source (not shown) to 300°C-400°C to sterilize and de-heat blade 140. The heat source may use resistive heating to heat blade 140 or a mounting block (not shown) in contact with blade 140, or blade 140 may be heated without direct contact, such as with a laser heater. Blade 140 is partially cooled before moving blade 140 along the cutting plane between first portions 120a, 125a and second portions 120b, 125b of jaws 120, 125, thereby cutting tubes 150a, 150b.

[0136] 8G shows a cut through the clamping unit 105 seen from above after the blade 140 has moved along the cutting plane to cut the tubes 150a, 150b. As a result, the first tube 150a is cut into a first portion 150a that connects to its respective consumable 13 and a second portion 150a' that is pre-routed to the sealed end of the tube 150a. Similarly, the second tube 150b is cut into a first portion 150b that connects to its respective consumable 13 and a second portion 150b' that is pre-routed to the sealed end of the tube 150b.

[0137] In FIG. 8H, the first portion 120a, 125a of the clamping unit 105 is moved relative to the second portion 120b, 125b of the clamping unit 105 to align the portions of the first tube 150a and the second tube 150b that connect to their respective consumables 13. The blade 140 remains between the first tube 150a and the second tube 150b and transfers heat from a heat source to melt the ends of the tubes 150a, 150b. The blade 140 may be held between the tubes 150a, 150b for a predetermined time or may have a predetermined thermal profile. In this example, the first portion 120a, 125a translates relative to the second portion 120b, 125b of the clamping unit 105 to align the tubes 150a, 150b, but it should be understood that alignment may be accomplished in other ways, such as by rotating one portion of the clamping unit 105 relative to the other portion. An infrared camera or infrared laser may be used in a closed loop to verify that the ends of the tubes 150a, 150b have reached the correct temperature for welding and that a uniform temperature has been reached. Alternatively, a thermistor, thermocouple, or resistance temperature detector (RTD) may be attached to a component such as the blade 140, a mounting block, or a heat source to monitor the temperature.

[0138] 8I, the blade 140 is removed from between the first and second portions 120a, 125a, 120b, 125b of the jaws 120, 125, and the clamping unit 105 brings the two tubes 150a, 150b into contact by translating the first and second portions 120a, 125a, 120b, 125b towards each other. The heat previously transferred to the tubes 150a, 150b by the blade 140 welds the tubes 150a, 150b together.

[0139] In FIG. 8J, the first jaw 120 and the second jaw 125 of the clamping unit 105 are moved apart to unclamp the tubes 150a, 150b, which are now connected to each other to form a single tube 150.

[0140] In Fig. 8K, a camera 160 is used to inspect the connection between two tubes 150a, 150b. The camera 160 has a microscope lens and is connected to a processing unit (not shown) that identifies whether the weld is successful or not, and the camera 160 may detect infrared (IR) radiation. The precession wheels 135a, 135b may be rotated to apply a tensile force to the tube 150 in the direction of the arrow, and the stress-strain profile of the tube 150 may be measured simultaneously. The stress-strain profile may also be analyzed by the processing unit to confirm whether the weld is successful or not.

[0141] Other mechanical tests may be used, such as, for example, torsion or vibration tests. Ultrasonic or x-ray sources may be used to test the presence of cavities in the connection. Fluid may also be pumped through the tube 150 and the camera 160 may be used to detect the presence of leaks. Alternatively, the connection may be placed in a sealed container with a pressure sensor that indicates a leak by detecting a pressure change in the container, or an odor detector may be used to measure a change in the concentration of water in the air. Alternatively, an external air pressure may be provided to the sealed container and the camera 160 may observe if air leaks into the connection. Alternatively, air may be pumped into the tubes 150a, 150b before welding and then a vacuum may be applied in the sealed container to see if air is leaking out. A biocompatible die may be added to the outside of the weld. If the processing unit determines that the weld was not successful, the tube 150 may be re-clamped and re-welded. An inspection of the connection may be performed before the tubes 150a, 150b are released by the clamping unit 105. By keeping the tubes 150a, 150b pinched during testing, even if there is a leak in the connection, the contents of the consumable 13 will still remain isolated from the surrounding air and atmosphere.

[0142] In FIG. 8L, the tube 150 is still pinched at the connection point, leading to a kink that prevents fluid flow through the tube 150. To untwist the tube 150, the tube 150 is manipulated perpendicular to the direction in which the tube 150 was pinched by the clamping unit 105 to open the tube 150 to allow fluid flow through the connection point. The manipulation may be performed by one of the gripping units 110a, 110b or by a separate gripping unit attached to the robot arm 3. Alternatively, the gripping units 110a, 110b may rotate the tube 150 by 90° inside the clamping unit 105 and partially re-clamp the tube 150 to remove the kink. There are other ways to open the tube 150, such as by applying a vacuum to the outside of the tube 150, or by embedding a magnet in the tube 150, by welding a spring to the tube 150 to open the tube 150 when the clamping unit 105 releases the tube 150, or by embedding a shape memory device in the tube 150 that can be actuated to change the shape of the tube 150, etc. The aforementioned inspection process may also be performed after the tubes 150a, 150b are opened, which allows for a good functional test of the tubes 150a, 150b. Preferably, the inspection process is performed both before the tubes 150a, 150b are released by the clamping unit 105 and after the tubes 150a, 150b are opened to allow for fluid flow.

[0143] With the connection between original tubing 150a, 150b complete, pumping unit 30 can be operated to pump fluid through tubing 150 between consumables 13 to perform a step of a cell therapy process.

[0144] Next, the separation process of the two consumables 13 will be described with reference to FIGS. 9A to 9D.

[0145] In FIG. 9A, the jaws 120, 125 of the clamping unit 105 are closed to pinch the tube 150 connecting between two consumables 13 (not shown). The clamping unit 105 for separating the consumables 13 may be located on a different end effector 100 than the clamping unit 105 used to connect the consumables 13. In FIG. 9B, the blade 140 is heated to 300° C.-400° C. by a heat source (not shown) to sterilize and / or deheat the blade 140. To obtain a good seal on the tube, the heating profile used during separation may be different from the heating profile used during connection. The blade 140 may be partially cooled. In FIG. 9C, the blade 140 is moved along the cutting plane between the first portion 120a, 125a and the second portion 120b, 125b of the jaws 120, 125, thereby cutting the tube 150 into a first tube 150a and a second tube 150b, each connected to a respective consumable 13. The blade 140 remains between the tubes 150a, 150b for a predetermined time and melts the ends of the tubes 150a, 150b. The predetermined time taken to melt the ends of the tubes 150a, 150b during separation may be different from the predetermined time taken to melt the ends of the tubes 150a, 150b during connection. In FIG. 9D, the blade 140 is removed from between the first portion 120a, 125a and the second portion 120b, 125b of the jaws 120, 125 of the clamping unit 105. Jaws 120, 125 of clamping unit 105 are opened to release tubes 150a, 150b from clamping unit 105. Gripper units 110a, 110b can now release tube holders 130a, 130b or manipulate tube holders 130a, 130b to attach one or both of tubes 150a, 150b to a different tube 150 that connects to a separate consumable 13 for a subsequent step in the cell therapy process.

[0146] If the end effector 100 for separating the consumables 13 is a different end effector 100 than the one for connecting the consumables 13, a different heat source and / or cutting method may be used. For example, an electromagnetic (EM) source, such as an RF source, may be used to seal the tubes 150a, 150b during the separation process. The EM source may be located on a separate robotic arm 3 or on a different robotic device 2. In this way, the tube 150 is clamped and fluid-sealed by the end effector 100 on the first robotic arm 3, and an EM source may be used to melt through the clamped portion of the tube 150. The EM source may have electrodes driven with an alternating current in the MHz or GHz frequency range, preferably 40.68 MHz. This melts the inside of the tube 150, sealing it extensively. A blade 140 may then be used to cut the heat-affected zone and separate the tube 150 into two tubes 150a, 150b.

[0147] 10-14, various alternative "non-contact" methods for sterilely connecting and disconnecting the two tubes 150a, 150b will now be described.

[0148] FIG. 10A shows two tubes 150a, 150b being joined together, each tube 150a, 150b terminating in a closed end. In FIG. 10B, the heat source applies heat directly to the closed ends of the tubes 150a, 150b, as indicated by the arrows. The heat source may be an electromagnetic radiation source, such as a laser that transmits light at infrared or radio frequencies. The material forming the tubes 150 may contain additives to improve absorption of the laser energy, or the tubes 150 may be painted with a material that absorbs the laser light. Preferably, the additive allows for two-photon polymerization, which requires high laser intensity for any activation, and allows the laser light to be nonlinearly focused on the welded ends of each tube 150, even when shown from the outside. Alternatively, the heat source may be an ultrasonic source, or the tubes 150 may contain additives that cause them to heat up during the use of an induction heater. Any of the above heating methods, including the use of wires or blades 140, may be used in combination.

[0149] In Fig. 10C, the heated ends of tubes 150a, 150b are pressed together such that tubes 150a, 150b are welded together to form a single tube 150. This may be accomplished by using the precession wheels 135a, 135b previously described, or by directly manipulating tubes 150a, 150b with gripping units 110a, 110b. In Fig. 10D, kinks in the connection of tubes 150 are removed using methods similar to those previously described. Inspection and quality control steps similar to those already described may also be applied using this process.

[0150] FIG. 11A shows the tube 150 separated into two parts. In FIG. 11B, the tube is pinched inside the clamping unit 105 as before, with the first jaw 120 of the clamping unit 105 split into a first part 120a and a second part 120b, and the second jaw 125 of the clamping unit 105 split into a first part 125a and a second part 125b. When the tube 150 is clamped between the first jaw 120 and the second jaw 125, a space remains between the first part 120a, 125a and the second part 120b, 125b of the jaws 120, 125. In FIG. 11C, a heat source applies heat directly to the tube 150 between the first part 120a, 125a and the second part 120b, 125b of the clamping unit 105, as indicated by the arrows. As a result, the heat source severs the tube 150 into a first portion 150a and a second portion 150b. In Figure 11D, the heat source continues to apply heat to the tube to ensure that the cut ends of the tubes 150a, 150b are sealed from the surrounding air.

[0151] FIG 12A shows two tubes 150a, 150b connected together via flanges 151a, 151b formed at each end. In the configuration shown, the two tubes 150a, 150b are joined together using the methods previously described. In FIG 12B, the tubes are joined, welded or clamped together at the location marked 170 to form a seal at the location marked 175. The tubes 150a, 150b can be heated using any of the methods previously described to weld them together. Methods of forming the flanges will now be described.

[0152] FIG. 13A shows a tube 150. In FIG. 13B, the tube 150 is mechanically pinched using a clamp 180 to prevent fluid flow through the tube 150. A pumping unit 30 can pump the fluid out of the clamp. In FIG. 13C, the tube 150 is forced into a heated die 190 to reform the tube 150 into the desired flange 151. In FIG. 13D, the die 190 can be cooled and separated into multiple portions 190a, 190b, 190c to release the flange 151 of the tube 150 from the die 190. The die 190 can be sterilized between each flange forming operation by autoclaving or by heating the die to an elevated temperature, such as a temperature above 400° C.

[0153] FIG. 14 shows another apparatus for forming a sterile weld using steam sterilization and heat welding. As well as many of the components already described in detail, the apparatus comprises a steam chamber 200 having a steam inlet 205. The tubes 150a, 150b to be welded together extend into the steam chamber 200 and may be open ended, but are pinched closed upstream, such as using a clamp or pump as already described. Steam is injected into the steam inlet 205 to sterilize the tubes 150a, 150b and the interior of the steam chamber. The heat from the steam also melts the ends of the tubes 150a, 150b that extend into the steam chamber 200. The precession wheels 135a, 135b then rotate to press and weld the ends of the tubes 150a, 150b together. Once welded, the tubes 150a, 150b are unpinched upstream and the pumping unit 30 can pump fluid through the connected tubes 150a, 150b.

[0154] FIG. 15A shows two tubes 150a, 150b being connected together with an alternative method for forming a sterile weld, where material is added to the tubes 150a, 150b. The tubes 150a, 150b may be cut using the methods described above or pinched upstream to prevent fluid flow. In FIG. 15B, once the tubes 150a, 150b are brought together, a hot material 210 is injected around the ends of the two tubes 150a, 150b. The material 210 is applied to the outside of the joint between the two tubes 150a, 150b using an injection mold (overmolding). This may be performed using a UV-curable adhesive or a heat-shrink adhesive.

[0155] FIG. 16A shows two tubes 150a, 150b having flanges 151a, 151b similar to those described in connection with FIGS. 12A, 12B, and 13A-13D. Here, the tubes 150a, 150b can be aseptically clamped together using the clamp unit 105 without the need for a heat source or welding. Heat may be used to sterilize the flanges 151a, 151b or to provide additional welding. In FIG. 16A, the first portion 120a, 125a of the clamp unit 105 clamps the first tube 150a, pinching the tube 150a adjacent to the flange 151a. Similarly, the second portion 120b, 125b of the clamp unit 105 clamps the second tube 150b, pinching the tube 150b adjacent to the flange 151b. 16B, the first portions 120a, 125a of the clamping unit 105 move towards the second portions 120b, 125b of the clamping unit 105 to pinch together the flanges 151a, 151b of the respective tubes 150a, 150b. Both portions 120a, 120b of the first jaw 120 then move away from both portions 125a, 125b of the second jaw 125, thereby unpinning the tubes 150a, 150b and providing a continuous tube 150 through which fluid can be pumped by the pumping unit 30.

[0156] FIG. 17A shows two tubes 150a, 150b connected together, with the ends of each tube 150a, 150b placed within a respective bag 220a, 220b. In FIG. 17B, the bags 220a, 220b are brought together and heat is applied to weld and melt the bags 220a, 220b together. Heat may be applied using any of the methods previously described. In FIG. 17C, a laser is used to cut a slot 230 connecting the two bags 220a, 220b, and in FIG. 17D, the tubes 150a, 150b within the bags 220a, 220b are brought together to form a connection. A laser may also be used here to weld the tubes 150a, 150b together. In Fig. 17E, after the tubes 150a, 150b are separated, the reverse process occurs where the tubes 150a, 150b are separated, and in Fig. 17F, the slot 230 between the bags 220a, 220b is welded together again. Now, the two bags 220a, 220b may be separated.

[0157] FIG. 18A shows an alternative apparatus for making a sterile connection comprising a sterilization box 230. The sterilization box 230 provides a localized sterile environment meaning that the tubes 150a, 150b may be connected to each other using a standard connection such as the connector 245 shown in FIG. 18B. The tubes 150a, 150b each have a respective duckbill valve 240a, 240b that is closed unless fluid is being pumped by the pumping unit 30. To operate the apparatus, the tubes 150a, 150b are inserted into the sterilization box 230, thereby sterilizing the tubes 150a, 150b. The tubes 150a, 150b are then connected together using the connector 245 immediately after sterilization has been performed by the sterilization box 230. The sterilization box 230 may be an autoclave box that sterilizes the tubes 150a, 150b using steam. Alternatively, the sterilization box 230 may use other methods such as ethanol sterilization (EtOH), ethylene oxide sterilization (EtO), gamma irradiation, UV sterilization, e-beam sterilization, or any combination of the above.

[0158] 19 shows an alternative apparatus for making a sterile connection, comprising a "T" connector 250 having a valved inlet 255 for steam. The interconnected tubes 150a, 150b each have a valve 260a, 260b that remains closed unless fluid is being pumped by the pumping unit 30. After connecting the tubes 150a, 150b to the T connector 250 while the valves 260a, 260b are closed, steam is pumped through the inlet 255 to sterilize the surface before the valves 260a, 260b are opened to allow fluid to flow through the tubes 150a, 150b.

[0159] 20 shows a needleless connector 270 having a first portion 270a and a second portion 270b that can be secured together to form a reversible connection. Prior to attachment, the first portion 270a and the second portion 270b of the needleless connector are sterilized by using an autoclave or a laser, or by using a high temperature blade 275 as shown herein.

[0160] 21 shows two tubes 150a, 150b each with a respective septum seal 280a, 280b. The tubes 150a, 150b can be connected together with a needle 290 that is first heated for sterilization and then inserted through the septum seals 280a, 280b.

[0161] FIG. 22A shows an alternative device for making a sterile connection, in which a blade 140 is attached to a surface 400. The surface 400 can be located in one of the processing stations 20. The device has a first jaw 120 divided into first and second portions 120a, 120b and a second jaw 125 divided into first and second portions 125a, 125b. The first portion 120a, 125a of the jaw is independently movable relative to the second portion 120b, 125b of the jaw, such as by use of one or more robotic arms 3 on a robotic device 2. In FIG. 22B, a first tube 150a is pinched by the first portion 120a, 125a of the jaw and a second tube 150b is pinched by the second portion 120b, 125b of the jaw. In Fig. 22C, a first tube 150a is cut by moving a first portion 120a, 125a of the jaws to the blade 140 on the surface 400. In Fig. 22D, a second tube 150b is cut by moving a second portion 120b, 125b of the jaws to the blade 140 on the surface 400. In Fig. 22E, the tubes 150a, 150b are moved to a heat source 410, which may be located in a separate processing station 20 from the blade 140 or in the same processing station 20 as the blade 140. The heat source 410 melts the ends of the tubes 150a, 150b that were cut by the blade 140. In Figure 22F, the first and second portions 120a, 125a, 120b, 125b of the jaws are moved together to contact the ends of the tubes 150a, 150b cut by the blade 140, thereby welding the tubes 150a, 150b together to form a single tube 150. In Figure 22G, the tube 150 is released by moving the first and second portions 120a, 120b of the first jaw 120 away from the first and second portions 125a, 125b of the second jaw 125.

[0162] FIG. 23A shows an alternative apparatus for making a sterile connection. The apparatus is similar to that shown in FIGS. 8A-8L, except that a clamping unit 105 is attached to a surface 420, such as the surface of one of the processing stations 20. The apparatus also has an end effector 100 for manipulating the tubes 150a, 150b via the holders 130a, 130b. In FIG. 23B, the end effector 100 moves a first tube 150a into the clamping unit 105, and in FIG. 23C, the end effector 100 moves a second tube 150b into the clamping unit 105. Two end effectors 100 may be used to move both tubes 150a and 150b into the clamping unit 105 simultaneously. In FIG. 23D, the first portion 120a, 120b of the first jaw 120 moves toward the second portion 125a, 125b of the second jaw 125 to clamp and pinch the tubes 150a, 150b to the clamping unit 105. In FIG. 23E, the blade 140 cuts the tubes 150a, 150b. As previously described, the blade 140 is heated before cutting the tubes 150a, 150b, and the heat melts the ends of the tubes 150a, 150b. In FIG. 23F, the blade 140 is removed and the first portion 120a, 125a of the clamping unit 105 moves relative to the second portion 120b, 125b of the clamping unit to align the tubes 150a, 150b leading to the respective consumables 13 (not shown). When the tubes 150a, 150b come into contact, they weld together to form a single tube 150. In Fig. 23G, the first and second portions 120a, 120b of the first jaw 120 move away from the first and second portions 125a, 125b of the second jaw 125 to release the tube 150. In Fig. 23H, the end effector 100 holds one of the holding portions 130a, allowing the tube 150 to be removed from the clamping unit 105. The tube 150 may then be placed into the pumping unit 30 so that fluid may be pumped through the tube 150.

[0163] FIG. 24A shows an alternative apparatus for making a sterile connection between containers 13a, 13b that are connected together using first and second portions 440a, 440b of a sterile connector 440 that are each attached directly to the respective container 13a, 13b. The robotic arm 3 is used to hold and manipulate each of the containers 13a, 13b. These connectors may be any reversible sterile connector 440, such as those described in FIG. 20, or a connector with an elastomeric seal. In FIG. 24B, the containers 13a, 13b are connected together using first and second portions 440a, 440b of the sterile connector 440, thereby forming a pathway for fluid between the containers 13a, 13b. In FIG. 24C, the robotic arm 3 tilts the containers so that under gravity, fluid can flow from the container 13a through the sterile connector and into the container 13b. The containers 13a, 13b may have a sterile air filter (not shown) that allows air to fill the containers 13a, 13b as fluid is transferred. Alternatively, a collapsible container such as a bag may be used.

[0164] It will be understood that other reversible connections known in the art may be adapted for use within the bioprocess (cell therapy) system 1. Such connections may be adapted to have features that are easily handled by the robotic device 2, such as a magnetic collar for easy alignment. It will be understood that any feature of a particular embodiment described herein may be applied to another embodiment in any suitable combination. It will also be understood that a particular combination of the various features described and defined in any aspect described herein may be implemented and / or provided and / or used independently. Any apparatus function described herein may be incorporated as a method function and vice versa.

[0165] Referring to FIG. 1, the reference numbers indicate the following features: 1000: Cell washing and concentration 1001: Cell raw materials 1002:Buffer 1003: Cell washer 1004: Waste 1005: Intermediate materials 1010: Activation, transduction, proliferation 1011: Reagents 1012: Culture medium 1013: Growth chamber 1014: Waste 1015: Collection 1020: Filling and finishing 1021: Final formulation 1022: Mixing chamber 1023: Product bag 1 1024: Product bag 2 1025: Product bag 3 1026: QC Bag

[0166] While the above is directed to exemplary embodiments of the invention, it will be understood that the invention is described herein by way of example only, and that detailed modifications may be made within the scope of the invention. Moreover, those skilled in the art will understand that the invention need not be limited to the embodiments disclosed herein, or to any details shown in the accompanying drawings that are not described in detail herein or defined in the claims. Indeed, such unnecessary features may be removed from the drawings without detracting from the invention.

[0167] Moreover, other and further embodiments of the invention will be apparent to those skilled in the art from consideration of this specification and can be devised without departing from the basic scope thereof, as determined by the following claims. [Explanation of symbols]

[0168] 1 bioprocessing system, 1a automation system, 2 robotic device, 2a base unit, 3 robot arm, 3-1 first robot arm, 3-2 second robot arm, 3a tool holder, tool belt, 4 thawing station, 6 centrifuge, 8 magnetic cell separator, 10 controlled rate freezer, 11 refrigerator, 12 incubator, 13 consumables, 13a container, 13b container, 13-1 identification mark, 14 housing, 15 user interface, 16 loading hatch, 17 factory floor, 18 rails, 19 tube supply means, 20 processing station, 30 pumping unit, 31 rotating wheel, 32 clamp, 35 observation system, 38 processing and control unit, 40 flange area, series of protrusions, handling section, 41 tube clip, 41-1 identification mark, 41-2 clip jaw, 50 gripping unit, 55 gripping part, 100 End effector, 100-1 First end effector, 100-2 Second end effector, 100a First part, 100b Second part, 105 Clamp unit, 110a Grip unit, 110b Grip unit, 120 First jaw, 120a First part, 120b Second part, 125 Second jaw, 125a First part, 125b Second part, 130a Tube holder, 130b Tube holder, 135a Precession wheel, 135b Precession wheel, 140 Blade, 150 Tube, 150a Tube, 150b Tube, 150-1 Identification mark, 150a' Second part, 150b' Second part, 151 Flange, 151a Flange, 151b Flange, 160 Camera, 190 Die, 190a Die, 190b Die, 190c Die, 200 Steam chamber, 205 Steam inlet, 210 Material, 220a Bag, 220b Bag, 230 Slot, Sterilization box, 240a Duckbill valve, 240b Duckbill valve, 245 Connector, 255 Inlet, 250 T-connector, 260a Valve, 260b Valve, 270 Needleless connector, 270a Needleless connector, 270b Needleless connector, 275 High temperature blade, 280a Septum seal, 280b Septum seal, 290 Needle, 300 Consumables, 301 Inlet / outlet, 310 Consumables, 311inlet / outlet, 312 sterile air filter, 313 vacuum-activated valve, 320 consumables, 321 inlet, 322 outlet, 323 inlet / outlet, 324 gas-permeable membrane, 325 cells, 326 medium, 400 surface, 410 heat source, 420 surface, 440 sterile connector, 440a sterile connector, 440b sterile connector

Claims

1. ・A series of processing stations for performing operations for a bioprocess, ・An automation system, comprising: wherein the automation system - means for manipulating a fluid connection between a first container and a second container separable therefrom, thereby creating a sterile connection that enables controlled transfer of fluid or cell material between the first container and the second container; the means for manipulating the fluid connection is configured to create a sterile connection separable after the transfer of fluid or cell material is completed so that the means can manipulate such a further fluid connection between the first container and a third container separable therefrom; - means for controlling an automated series of operations of the processing stations A bioprocess system comprising.

2. The bioprocess system according to claim 1, wherein the means for manipulating the fluid connection is further configured to seal the separated fluid connection so that the transfer of fluid or cells to or from the first and second containers is blocked.

3. The bioprocess system according to claim 1, further comprising installing one or more of the containers at each of the series of processing stations and means for moving the containers between stations.

4. The bioprocess system according to claim 1, further comprising means for enabling the transfer of fluid or cells between aseptically connected containers.

5. The bioprocess system according to claim 1, further comprising means for determining whether the fluid connection is properly connected, for example by inspecting the fluid connection.

6. The bioprocess system according to claim 5, further comprising an image capture device or device such as a camera for inspecting the sterile connection between the tubes and / or for identifying each of the containers.

7. The bioprocess system according to claim 1, wherein the system is configured to process a plurality of containers simultaneously.

8. The bioprocess system according to claim 7, wherein the means for controlling an automated series of operations is configured to automatically schedule a series of actions followed by the bioprocess system.

9. The bioprocess system according to claim 1, wherein the means for operating the fluid connection is configured to create a sterile connection between a first tube connected to the first container and a second tube connected to the second container.

10. The bioprocess system according to claim 9, wherein the means for operating the fluid connection comprises a tube welder configured to join the first tube to the second tube.

11. The bioprocess system according to claim 10, further comprising means for manipulating the joined tubes to release the pinched portion, thereby establishing a fluid path through the joined tubes.

12. Further comprising at least one tube clip, The bioprocess system according to claim 9, wherein the tube clip is configured to hold the first or second tube in a fixed position relative to the tube clip.

13. An automated system for fluidly connecting two containers, wherein at least a first container has a tube fluidly connected to a first end, and a second end of the tube is configured to form a sterile connection with another such tube, The automated system is A robotic device configured to engage and position the tube at one or more positions where the second end of the tube fluidly connected to the first container is operated, Means for manipulating a portion of the tube toward the second end of the tube, thereby configuring the second end of the tube to create a sterile connection with another such tube, An automated system comprising.

14. The means for manipulating a portion of the tube is Means for clamping a portion of the tube toward the second end of the tube, thereby forming a pinched portion in the tube, whereby the tube is fluidly sealed upstream of the pinched portion, Means for removing a section of the tube downstream of the pinched portion, thereby removing the second end of the tube, whereby a new second end of the tube that has not previously contacted another such tube is formed, The automated system according to claim 13, further comprising.

15. The automated system according to claim 14, further comprising means for enabling a controlled transfer of fluid and cell material between the first container and the second container.

16. (i) said means for clamping a portion of the tube; (ii) said means for removing a section of the tube; The automated system according to claim 14, wherein at least one of them is configured as an end effector for a robotic arm.

17. The automated system according to claim 14, further comprising means for releasing the picked portion when joined to another such tube, thereby manipulating the tube to establish a fluid path through the joined tubes.

18. The automated system according to claim 16, wherein the end effector comprises at least one gripping unit configured to engage the tube and move the tube.

19. A method of performing a bioprocess in a system having a series of processing stations for performing operations for a bioprocess using a plurality of containers, wherein the method - configuring an automated system to manipulate a fluid connection between a first container and a separable second container, thereby creating a sterile connection that enables a controlled transfer of fluid or cell material between the first container and the second container; a step, wherein the step of manipulating the fluid connection creates a sterile connection that can be separated after the transfer of fluid or cell material is complete, such that a further such fluid connection between the first container and a separable third container can be manipulated; a step, - controlling an automated series of operations of the processing stations; A method comprising.

20. The method according to claim 19, further comprising the step of controlling the automated series of operations according to a predetermined workflow.

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