Automated Hollow Fiber System

Abstract

A computer-implemented method that, when executed on the data processing hardware, causes the data processing hardware to perform operations on a hollow fiber transduction device, including directing a pump of the hollow fiber transduction device to perform one of the steps of the recipe that provides a first pressure specified in the recipe to a first sensor, including receiving a fluid flow signal from the first sensor of the hollow fiber transduction device indicative of an operating parameter measured at the first sensor, comparing a first termination trigger threshold of one of the steps of the recipe to the received fluid flow signal, and directing the pump to complete one of the steps of the recipe based on the comparison of the first termination trigger threshold.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 287,651, filed December 9, 2021, and U.S. Provisional Application No. 63 / 365,829, filed June 3, 2022. The disclosures of these prior applications are considered part of the disclosure of this application and are incorporated by reference herein in their entireties.

[0002] The present disclosure relates to an automated hollow fiber system for the transduction of cells. [Background technology]

[0003] Cell therapy exploits natural transduction processes and typically uses viral particles modified for safety and functionality as a delivery vehicle (vector) to introduce genetic material into a patient's cells or donor's cells, for example, for the manufacture of autologous or allogeneic cell therapies. Viral vector transduction is currently the most frequently used method in cell therapy manufacturing to introduce therapeutic genetic material into cells (such as immune cells or stem cells). Current transduction processes employed in cell therapy manufacturing tend to be labor-intensive and inefficient, especially at large scale, in the use of viral vectors, which increases the manufacturing costs of cell therapies and the time required to produce such therapies. Thus, there is a need in the industry for more efficient and cost-effective viral transduction processes for the manufacture of cell therapies, especially ex vivo cell therapies. Summary of the Invention

[0004] The present disclosure provides automated hollow fiber systems and methods for transduction of cells for cell therapy manufacturing. The methods and systems described herein are less labor intensive, more cost effective and efficient, and suitable for large scale manufacturing of cell therapies. Furthermore, the automated hollow fiber systems and methods described below minimize the risk of contamination during cell therapy manufacturing by automating the transduction process, resulting in more consistent and uniform cell transduction.

[0005] One aspect of the disclosure provides a computer-implemented method that, when executed on data processing hardware, causes the data processing hardware to perform operations on a hollow fiber transduction device. The hollow fiber transduction device includes a pump and a plurality of sensors for executing a recipe including a plurality of steps, where each step includes a first termination trigger threshold indicating whether a respective step of the recipe is completed. The operations include directing the pump to execute one of the steps of the recipe to provide a first pressure at a first sensor of the plurality of sensors. The first pressure is specified in the recipe. The operations also include receiving a fluid flow signal from the first sensor of the plurality of sensors indicative of an operational parameter measured at the first sensor. The operations also include comparing a first termination trigger threshold for one of the steps of the recipe to the received fluid flow signal. The comparison of the first termination trigger threshold indicates whether an operational condition is satisfied. The operations also include directing the pump to complete one of the steps of the recipe based on the comparison of the first termination trigger threshold.

[0006] Implementations of the present disclosure may include one or more of the following optional features: In some implementations, each step of the recipe includes at least one of a valve configuration, a pump direction, a pump speed, or a step type. In some examples, directing the pump to perform one of the steps of the recipe that provides a first pressure at a first sensor of the plurality of sensors includes sending a pump speed signal to the pump to operate at a first pump speed, where directing the pump to perform one of the steps of the recipe that provides a first pressure at a first sensor of the plurality of sensors further includes sending a pump direction signal to the pump to operate in a first direction. In these examples, after directing the pump to complete one of the steps of the recipe based on a comparison of the first termination trigger threshold, the operation may further include directing the pump to perform one of another steps of the recipe that provides a second pressure specified in the recipe at a first sensor of the plurality of sensors, where one of the other steps of the recipe further directs the pump to operate in a second direction opposite the first direction.

[0007] In some implementations, the operations further include, if the received fluid flow signal does not meet a first termination trigger threshold for one of the recipe steps, waiting a time corresponding to a timer value, receiving a second fluid flow signal from a first sensor of the plurality of sensors indicative of an operating parameter measured by the first sensor, comparing the first termination trigger threshold for one of the recipe steps to the received second fluid flow signal, and in response to the comparison, instructing the pump to re-execute one of the recipe steps. In these implementations, the operating parameter includes at least one of a pressure value or a fluid flow value.

[0008] The operations may further include determining whether one of the steps of the recipe is the last step of the recipe. In some examples, the operations further include completing the execution of the recipe if one of the steps of the recipe is the last step of the recipe. In other examples, the operations further include executing the next step of the recipe if one of the steps of the recipe is not the last step of the recipe. One of the steps of the recipe may include a second termination trigger threshold indicating whether a fault occurred in the hollow fiber transduction device during execution of the respective step of the recipe. Here, each of the first termination trigger threshold and the second termination trigger threshold includes at least one of a pressure value, a timer value, an air volume value, a fluid flow value, or a fluid to air ratio. In some examples, the operations further include comparing a second termination trigger threshold for one of the steps of the recipe to the received fluid flow signal, and instructing the pump to ignore one of the steps of the recipe based on the comparison of the second termination trigger threshold.

[0009] Optionally, the operations may further include storing the recipe in memory hardware in communication with the data processing hardware. In some examples, the operations further include receiving a request to execute the recipe stored in the memory hardware, reading the recipe stored in the memory hardware, and executing the recipe by the data processing hardware. In some implementations, the operations further include receiving a request to modify at least one step of the recipe stored in the memory hardware, and updating the at least one step of the recipe stored in the memory hardware.

[0010] Another aspect of the present disclosure provides a system including a filter module defining an intracapillary space and an extracapillary space separated from the intracapillary space by a porous membrane, where the filter module includes a pair of intracapillary ports fluidly coupled to the intracapillary space and a pair of extracapillary ports coupled to the extracapillary space. The system also includes a first pump including a first pump inlet in communication with each of a cell source having cells and a vector source having vectors, and a first pump outlet in selective communication with each of the pair of intracapillary ports and the pair of extracapillary ports. The system also includes a second pump including a second pump inlet in communication with each of the cell source and the vector source, and a second pump outlet in selective communication with each of the pair of intracapillary ports and the pair of extracapillary ports. The system also includes a graphical user interface (GUI) in operative communication with each of the first pump and the second pump, and is configured to receive a request from a user to operate the first pump and the second pump according to a recipe including one or more steps.

[0011] Embodiments of the present disclosure may include one or more of the following optional features: In some embodiments, the system further includes at least one valve disposed between the first pump outlet and a first intra-capillary port of the pair of intra-capillary ports, at least one valve disposed between the first pump outlet and a second intra-capillary port of the pair of intra-capillary ports, and a fluid sensor disposed between the first pump outlet and the first intra-capillary port of the pair of intra-capillary ports, where the valves operate according to a recipe. In some examples, the system further includes a valve disposed between the first pump inlet and a cell source, and an air bubble sensor disposed between the first pump inlet and a cell source, where the valves operate according to a recipe.

[0012] The system may further include at least one valve disposed between the second pump outlet and a first intra-capillary port of the pair of extra-capillary ports, at least one valve disposed between the second pump outlet and a second intra-capillary port of the pair of extra-capillary ports, and a fluid sensor disposed between the second pump outlet and the first intra-capillary port of the pair of extra-capillary ports, where the valves operate according to a recipe. Optionally, the GUI is further configured to display the recipe from the memory hardware to the user in response to receiving a request.

[0013] Another aspect of the present disclosure provides a system including a filter module defining an intracapillary space and an extracapillary space separated from the intracapillary space by a porous membrane. The filter module includes a pair of intracapillary ports fluidly coupled to opposite ends of the intracapillary space and a pair of extracapillary ports coupled to opposite ends of the extracapillary space. The system also includes a first pump including a first pump inlet in communication with each of a cell source having cells and a vector source having vectors, and a first pump outlet in selective communication with a first intracapillary port of the pair of intracapillary ports and a second intracapillary port of the pair of intracapillary ports. The system also includes a first fluid sensor disposed between the first pump outlet and the first intracapillary port of the pair of intracapillary ports. The system also includes data processing hardware and memory hardware storing instructions that, when executed on the data processing hardware, cause the data processing hardware to operate according to a recipe for transducing cells with a vector.

[0014] Implementations of the present disclosure may include one or more of the following optional features. In some implementations, the operations include operating a first pump at a first pump rate to provide a first flow of at least one of cells or vectors to each of a pair of intracapillary ports of the filter module, obtaining a first fluid flow signal from a first fluid sensor indicative of the first flow of at least one of cells or vectors, comparing the first fluid flow signal to a first fluid flow parameter threshold, and operating the first pump at a second pump rate in response to the comparison. In these implementations, the operations may further include initiating a pump rate adjustment procedure in response to the comparison. The pump rate adjustment procedure includes ceasing operation of the first pump for a predetermined period of time. The predetermined period of time may be a fixed period of time. The predetermined period of time may be a pressure adjustment period. In some examples, the pump rate adjustment procedure further includes operating the first pump in a reverse flow condition.

[0015] In some embodiments, the operations further include operating the first pump at a third pump rate to provide a second flow of the other of the cells or vectors to each of the pair of intracapillary ports of the filter module. In these embodiments, the operations may further include directing the first pump to alternate between operating at the first pump rate to provide the first flow of at least one of the cells or vectors to each of the pair of intracapillary ports of the filter module and operating at a third pump rate to provide the second flow of the other of the cells or vectors to each of the pair of intracapillary ports of the filter module. The operations may further include operating the first pump at the first pump rate to provide the first flow of at least one of the cells or vectors to each of the pair of intracapillary ports of the filter module, and then operating the first pump at the third pump rate to provide the second flow of the other of the cells or vectors to each of the pair of intracapillary ports of the filter module. In some examples, the operations further include obtaining a second fluid flow signal from the first fluid sensor indicative of a second flow of one of the cells or vectors, comparing the second fluid flow signal to a second fluid flow parameter threshold, and initiating a pump rate adjustment procedure in response to the comparison.

[0016] In some examples, the system further includes a transduction medium source in communication with the first pump inlet, the transduction medium being included therein. In these examples, the operations further include operating the first pump at a third pump rate to provide a third flow of the transduction medium to each of the pair of intracapillary ports. The system may further include a second pump including a second pump inlet in communication with each of the cell source and the vector source, and a second pump outlet in selective communication with each of the pair of extracapillary ports. Here, the system may further include a culture medium source in communication with the first pump inlet and the second pump inlet, the culture medium being included therein. In some examples, the operations further include operating the first pump at a fourth pump rate to provide a fourth flow of the culture medium to one of the intracapillary ports, and operating the second pump at a fifth pump rate to provide a fifth flow of the culture medium to each of the pair of extracapillary ports.

[0017] In some embodiments, the vector used to transduce the cells includes a non-viral vector or a viral vector. In some examples, the system further includes a mixer configured to receive at least one of the cell population or the vector particles. In these examples, the operations further include mixing the cells and the vector using the mixer and then loading both mixtures into the filter module. These operations may further include loading the cell population and the vector particles into the intracapillary space, thereby transducing one or more of the cells in the cell population in the intracapillary space, thereby generating a cell therapy product including one or more transduced cells, and harvesting the cell population including the one or more transduced cells from the intracapillary space.

[0018] One aspect of the disclosure provides a computer-implemented method for manufacturing a cell therapy product, the method comprising, when executed on the data processing hardware, causing the data processing hardware to perform operations to execute a recipe including a plurality of steps in a hollow fiber transduction device. The hollow fiber transduction device includes a pump including a pump inlet in communication with each of a cell source having cells and a vector source having vectors, and a first pump outlet in selective communication with a first intracapillary port and a second intracapillary port of a filter module. The operations include directing the pump to execute one of the steps of the recipe to provide cells and vectors to the first intracapillary port and the second intracapillary port of the filter module to transduce the cells. The operations also include directing the pump to execute a subsequent step of the recipe to provide the transduced cells from the filter module to a collection container in communication with the second intracapillary port of the filter module.

[0019] Embodiments of the present disclosure may include one or more of the following optional features: In some embodiments, the operations further include growing the transduced cells in a suitable culture medium according to the steps of a recipe. In some examples, the operations further include cryopreserving the transduced cells after growing the transduced cells according to the steps of a recipe. In some embodiments, at least 50%, 60%, 70%, 80%, or 90% or more of the cells provided to the filter module are transduced. Optionally, the vector may include a viral vector or a non-viral vector. The vector may include a chimeric antigen receptor (CAR) construct expressed on the surface of the transduced cells. In some examples, the operations further include separating the transduced cells from untransduced cells according to the steps of a recipe.

[0020] The details of one or more embodiments of the disclosure are set forth in the accompanying drawings and the description below. Other aspects, features, and advantages will be apparent from the description and drawings, and from the claims. [Brief description of the drawings]

[0021] [Figure 1A] FIG. 1 shows a perspective view of a hollow fiber system. [Figure 1B] FIG. 1 shows a perspective view of a hollow fiber system. [Figure 1C] FIG. 1 shows a perspective view of a hollow fiber system. [Figure 1D] FIG. 2 is a schematic diagram of a controller and a graphical user interface in communication with the components of the hollow fiber system. [Diagram 2] 1 is a schematic diagram of an example computing device that can be used to implement the systems and methods described herein. [Figure 3A] FIG. 1 illustrates a schematic diagram of a hollow fiber system showing the fluid flow paths. [Figure 3B] 1 illustrates a horizontal cross section of a simplified example of a hollow fiber of a filter module. [Figure 3C] 1 illustrates a vertical cross section of a simplified example of a hollow fiber of a filter module. [Figure 3D] 1 illustrates a horizontal cross section of a filter module containing a plurality of hollow fibers. [Figure 4] 1 is a flow chart illustrating an example arrangement of method operations for performing graphical user interface operations in user mode and developer mode. [Figure 5A] FIG. 2 is a schematic diagram of an example recipe. [Figure 5B] FIG. 5B is a schematic diagram of an example of an end trigger threshold for the recipe of FIG. 5A. [Figure 6] 1 is a flow chart of an example implementation of the operations of a method for performing an initialization routine in a hollow fiber system. [Figure 7] 4 is a flowchart of an example of a method operation for executing steps of a recipe. [Figure 8] 11 is a flowchart of an example embodiment of a method of performing a pump rate adjustment procedure. [Figure 9] 1 is a flow chart of an example implementation of a method of operating a hollow fiber system according to a recipe. [Figure 10]11 is a flowchart of an example operation of a method for initiating a pump rate adjustment procedure while operating a pump at a first pump speed according to a recipe. [Figure 11] 11 is a flowchart of an example operation of a method for initiating a pump rate adjustment procedure while operating a pump with first fluid flow parameters according to a recipe. [Figure 12] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 13] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 14] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 15] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 16] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 17] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 18] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 19] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 20] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 21] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 22] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Diagram 23] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Figure 24] 1 illustrates a schematic diagram of an example of a fluid flow path during recipe execution. [Diagram 25] FIG. 1 illustrates a schematic diagram of a hollow fiber system showing the fluid flow paths. [Figure 26] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Figure 27] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Figure 28]26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Figure 29] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 30] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 31] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 32] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 33] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 34] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 35] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 36] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Figure 37] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Figure 38] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Figure 39] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 40] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 41] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 42] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 43] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 44] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Diagram 45] 26 illustrates a schematic diagram of an example of a fluid flow path through the hollow fiber system of FIG. 25 during recipe execution. [Figure 46] Illustrated is a cell and vector loading chart illustrating constant pressure vs. constant flow rate. [Figure 47] 1 illustrates a chart of hollow fiber transduction efficiency by length. [Figure 48] 1 illustrates a cell and vector loading chart, which illustrates a comparison of premix loading and sequential loading. [Figure 49] 1 illustrates a chart of hollow fiber transduction of natural killer cells. [Figure 50] Illustrated chart of optimization study in a fully automated platform demonstrating retroviral transduction of γδ T cells in a hollow fiber system. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] Like reference symbols in the various drawings indicate like elements.

[0023] Transduction is the process by which a virus infects a target or host cell. Viruses naturally undergo the transduction process and have evolved to be very efficient at transferring genetic material into target cells. For transduction to occur, the viral particle must come into physical contact with the target cell to first bind to, invade, and finally transfer genetic material into the target cell. Binding occurs through specific protein-protein interactions with the correct proteins required by both the virus and the target cell.

[0024] Cell therapy exploits the natural transduction process and uses viral particles modified for safety and function as delivery vehicles (vectors) to introduce genetic material into cells from a patient (e.g., in autologous cell therapy) or donor (e.g., in allogeneic cell therapy). Viral vector transduction is currently the method most frequently used in the manufacture of cell therapies to introduce genetic material into cells.

[0025] Current industry approaches to viral transduction include static transduction systems, the use of chemical enhancers, and spinoculation, each of which is further described below.

[0026] Viral transduction under static conditions is the most common method by which viral transduction is currently performed in industry. In standard static transduction methods, most transduction is performed in standard culture flasks or bags under static culture conditions. In such methods, the viral vector is suspended in the medium containing the cell population, and the transduction efficiency is mainly dependent on chance. Transduction using standard static methods faces various problems that result in inefficient transduction of cells. For example, the use of static methods results in the presence of small vector particles that remain in suspension and cannot reach the target cells. This occurs, at least in part, because larger cells quickly settle to the floor of the culture vessel. The net result of using static culture methods for transduction is that only a small fraction of the vector particles can reach the cells by diffusion alone. As a result, transduction efficiencies are low and large amounts of viral vector are required to achieve significant cell transduction. This is because the binding of the viral vector to the target cells depends on the expression of and physical contact with the receptor / ligand. Thus, the rate of transduction is proportional to the local concentration of the virus in a given cell. The need for large quantities of viral vectors to achieve sufficient transduction rates is costly and can create inefficiencies in the overall cell therapy manufacturing process.

[0027] Another standard method for transduction of cells is the use of spinoculation. Spinoculation refers to the inoculation of cells using centrifugal force. Spinoculation reduces the volume occupied by the cells. This technique has been shown to have various negative aspects, e.g. damage to the cells, difficulties in scaling up, and is generally less effective for small vectors.

[0028] Yet another method for improving the transduction efficiency of viruses, especially retroviruses, is through the use of cell adhesion substances or chemical enhancers that bind to retroviruses, such as fibronectin or fibronectin fragment CH-296 [Retronectin® (recombinant human fibronectin fragment) or retronectin]. This method requires adding a solution containing retroviral vectors to a vessel coated with retronectin, followed by incubation for a certain time that allows only viral vectors to bind to retronectin, and then removing the supernatant containing inhibitors for viral infection before adding target cells. Coating the vessel surface with retronectin is time-consuming, making this method expensive. Moreover, this method is difficult to scale up when a large number of cells need to be transduced.

[0029] The embodiments herein are directed to a fully automated hollow fiber system for performing cell transduction. In some embodiments, the transduction methods described herein eliminate the need for any chemical enhancers or cell adhesives. In other embodiments, the transduction methods described herein may also use chemical enhancers or cell adhesives for efficient transduction. In various embodiments described herein, cell transduction may be performed using viral or non-viral vectors.

[0030] 1A-1C illustrate perspective views of a fully automated hollow fiber cell transduction system 100. The fully automated hollow fiber cell transduction system 100 (also referred to herein simply as "hollow fiber system 100") includes one or more bag hooks 102, a hook adjustment handle 104, and a status light 106. Each bag hook 102 of the one or more bag hooks 102 is configured to support an input container 302 or an output container 322 (FIG. 3). The hook adjustment handle 104 is configured to adjust the bag hook 102 to vertically raise or lower the input container 302 or the output container 322. In some examples, the status light 106 indicates the operational status of the device to a user of the hollow fiber system 100. For example, the status light 106 may include a green light to indicate to a user that a process is active, a blue light to indicate that the process is complete, and a red light to indicate that user input is required.

[0031] The hollow fiber system 100 may also include one or more pumps 328, 332, an air bubble sensor 318, a fluid sensor 320, and a pinch valve 314, as described in more detail with reference to FIG. 3. Referring now to FIG. 1A, a perspective view illustrates the hollow fiber system 100, which also includes a first pump 328 including a first pump inlet 327 and a first pump outlet 329, a second pump 332 including a second pump inlet 331 and a second pump outlet 333, and a filter module 334, as described in more detail with reference to FIG. 3A. The hollow fiber system 100 is aseptically closed from the surrounding environment, i.e., the hollow fiber system 100 is free of bacteria or viruses on the exterior surface of the hollow fiber system 100 itself.

[0032] 1C illustrates an example of a tubing set 160 disposed on a mounting card 124 configured to mount to a hollow fiber system 100. That is, the post mounting holes 120 of the mounting card 124 are attached to the tubing set mounting posts 118 (FIG. 1A) of the hollow fiber system 100. The tubing set 160 also includes a number of conduits (e.g., transduction conduit 340, air filter conduit 342, cell conduit 344, culture conduit 346, first and second viral conduits 348a, 348b, waste conduit 350, harvest conduit 352, first and second intra-capillary conduits 356a, 356b, and first and second extra-capillary sub-conduits 368a, 368b) that fluidly couple the components of the hollow fiber system 100 to one another, as described in more detail below with respect to FIG. 3A. The conduits are PVC tubing and are adjustable to change the fluid path of the hollow fiber system 100. Specifically, the conduits 340, 342, 344, 346, 348, 350, 352, 356, 368 are attached to a plurality of tube holders 122, which allows a user to easily and quickly adjust the fluid path of the conduits. In some examples, the tube set 160 includes a fluid sensor 320 (e.g., a first fluid sensor 320, 320a and a second fluid sensor 320, 320b), a filter module 334, and an air filter 312. In these examples, the fluid sensor 320 may be a disposable fluid sensor. In some embodiments, once the cells 306 (shown in FIG. 3A) are harvested via the harvest conduit 352, the cells are transferred to an appropriate bioreactor or culture vessel for growth. The transduced cells are then grown in a suitable culture medium for 3-20 days, washed out, suspended in final formulation buffer, and stored frozen as a formulation suitable for future therapeutic use.

[0033] Optionally, hollow fiber cell transduction system 100 may also include an Ethernet port 116, a graphical user interface (GUI) 180, and a tubing set mounting post 118. Ethernet port 116 may communicatively couple hollow fiber system 100 to a remote computing system configured to execute instructions remotely on hollow fiber system 100. GUI 180 allows a user to provide user input that controls operations on hollow fiber system 100. Hollow fiber system 100 also includes a computing device 200 (FIG. 1B) in communication with each component of hollow fiber system 100. Computing device (i.e., controller) 200 may include a processor (e.g., data processing hardware) 210 and memory (e.g., memory hardware) 220, which will be described in more detail with reference to FIG. 2.

[0034] 1D illustrates electrical communication between the GUI 180, the processor 210, the memory hardware 220, and components of the hollow fiber system 100. In the illustrated example, the GUI 180 sends a request 182 to the processor 210 to execute a recipe 500 from a plurality of recipes 500, 500a-500n stored in the memory hardware 220 of the controller 200. Each recipe 500 of the plurality of recipes 500 stored in the memory hardware 220 includes a plurality of steps 510, 510a-510n, where each step 510 instructs the processor 210 to obtain information from and / or instruct components of the hollow fiber system 100. In response to receiving the request 182, the processor 210 sends a read recipe request 212 to the memory hardware 220 to read the recipe 500 corresponding to the read recipe request 182.

[0035] In some implementations, the controller 200 is in electrical communication with the air filter 312, the flow valve 314, the pinch valve 316, the air bubble sensor 318, the fluid sensor 320, and the pumps 328, 332. In the illustrated example, the processor 210 receives an air filter signal 211 from the air filter 312 and sends an air filter command signal 212 to the air filter 312. In this example, the processor 210 receives a flow valve signal 213 from the flow valve 314 and sends a flow valve command signal 214 to the flow valve 314. Here, the processor 210 also receives a pinch valve signal 215 from the pinch valve 316 and sends a pinch valve command signal 216 to the flow valve 314. Continuing with the example, the processor 210 receives an air bubble sensor signal 217 from the air bubble sensor 318 and sends an air bubble sensor command signal 218 to the air bubble sensor 318. The processor 210 also receives a fluid flow signal 219 from the fluid sensor 320 and sends a fluid flow command signal 221 to the fluid sensor 320. In some examples, the fluid sensor 320 is a pressure sensor and the fluid flow signal 219 indicates a pressure value. In other examples, the fluid sensor 320 is a fluid flow sensor and the fluid flow signal 219 indicates an amount of fluid flow (e.g., liters / minute) through the fluid sensor 320. In this example, the processor 210 also receives a pump signal 229 from the pumps 328, 332 and sends a pump command signal 231 to the pumps 328, 332. The contents of the command signals are described in further detail with reference to Figures 5A and 5B.

[0036] In some examples, the processor 210 reads the signals 211, 213, 215, 217, 219, 229 at predetermined time intervals. In other examples, the recipe 500 instructs the processor 210 when to read the signals 211, 213, 215, 217, 219, 229. The processor 210 sends the results 184 to the GUI 180 based on the execution of the recipe 500 of the request 182. The GUI 180 displays the results 184 to the user and may include information received from one of the components of the hollow fiber system 100. Thus, the hollow fiber system 100 is a closed system in that the controller 200 sends the command signals 212, 214, 216, 218, 221, 231 based, at least in part, on the read signals 211, 213, 215, 217, 229.

[0037] 2 is a schematic diagram of a computing device (i.e., controller) 200 that can be used to implement the systems and methods described in this disclosure. The components shown therein, their connections and relationships, and their functions are for illustrative purposes only and are not meant to limit the embodiments of the invention described and / or claimed in this disclosure.

[0038] Computing device 200 includes processor (e.g., data processing hardware) 210, memory (e.g., memory hardware) 220, storage device 230, high-speed interface / controller 240 connecting to memory 220, high-speed expansion port 250, and low-speed interface / controller 260 connecting to low-speed bus 270 and storage device 230. Each of components 210, 220, 230, 240, 250, and 260 are interconnected using various buses and may be mounted on a common motherboard or otherwise as desired. Processor 210 may process instructions (e.g., recipes) including instructions stored in memory 220 or storage device 230 for execution within computing device 200 to display graphical information for a graphical user interface (GUI) 180 on an external input / output device, such as a display 280 coupled to high-speed interface 240. In other implementations, multiple processors and / or multiple buses may be used along with multiple memories and types of memories as desired. Also, multiple computing devices 200 may be connected, with each device providing a portion of the required operations (eg, as a bank of servers, a group of blade servers, or a multi-processor system).

[0039] The memory 220 stores information non-transiently within the computing device 200. The memory 220 may be a computer-readable medium, a volatile memory unit, or a non-volatile memory unit. The non-transient memory 220 may be a physical device used to temporarily or permanently store programs (e.g., a sequence of instructions) or data (e.g., program state information) for use by the computing device 200. Examples of non-volatile memory include, but are not limited to, flash memory and read-only memory (ROM) / programmable read-only memory (PROM) / erasable programmable read-only memory (EPROM) / electronically erasable programmable read-only memory (EEPROM) (e.g., typically used for firmware such as boot programs). Examples of volatile memory include, but are not limited to, random access memory (RAM), dynamic random access memory (DRAM), static random access memory (SRAM), phase change memory (PCM), and disk or tape.

[0040] The storage device 230 can provide mass storage for the computing device 200. In some implementations, the storage device 230 is a computer-readable medium. In various different implementations, the storage device 230 can be a floppy disk drive, a hard disk drive, an optical disk drive, or a tape drive, a flash memory or other similar solid-state memory device, or an array of devices including devices in a storage area network or other configuration. In additional implementations, the computer program product is tangibly embodied in an information carrier. The computer program product includes instructions that, when executed, perform one or more methods, such as the methods described in this disclosure. The information carrier is a computer or machine-readable medium, such as the memory 220, the storage device 230, or a memory on the processor 210.

[0041] The high-speed controller 240 manages the bandwidth-intensive operations of the computing device 200, and the low-speed controller 260 manages the less bandwidth-intensive operations. Such an assignment of duties is merely an example. In some implementations, the high-speed controller 240 is coupled to the memory 220, the display 280 (e.g., via a graphics processor or accelerator), and is coupled to a high-speed expansion port 250 that can accept various expansion cards (not shown). In some implementations, the low-speed controller 260 is coupled to the storage device 230 and the low-speed expansion port 290. The low-speed expansion port 290, which may include various communication ports (e.g., USB, Bluetooth, Ethernet, wireless Ethernet), may be coupled, for example, via a network adapter, to one or more input / output devices, such as a keyboard, a pointing device, a scanner, or a networking device such as a switch or router.

[0042] Computing device 200 may be implemented in many different forms, as shown in the figure, for example as a standard server 200a, or multiple times within a group of such servers 200a, as a laptop computer 200b, or as part of a rack server system 200c.

[0043] Various implementations of the systems and techniques described herein may be realized in digital electronic and / or optical circuitry, integrated circuits, specially designed ASICs (application specific integrated circuits), computer hardware, firmware, software, and / or combinations thereof. These various implementations may include implementation in one or more computer programs executable and / or interpretable on a programmable system including at least one programmable processor, which may be special-purpose or general-purpose, coupled to receive or transmit data and instructions from or to the storage system, at least one input device, and at least one output device.

[0044] A software application (i.e., a software resource) can refer to computer software that causes a computing device to perform tasks. In some examples, a software application may be referred to as an "application," an "app," or a "program." Examples of applications include, but are not limited to, system diagnostic applications, system management applications, system maintenance applications, word processing applications, spreadsheet applications, messaging applications, media streaming applications, social networking applications, and gaming applications.

[0045] These computer programs (also known as programs, software, software applications, or code) include machine instructions for a programmable processor and may be implemented in high-level procedural and / or object-oriented programming languages, and / or assembly / machine languages. As used herein, the terms "machine-readable medium" and "computer-readable medium" refer to any computer program product, non-transitory computer-readable medium, apparatus, and / or device (e.g., magnetic disk, optical disk, memory, programmable logic circuit (PLD)) used to provide machine instructions and / or data to a programmable processor, including a machine-readable medium that receives machine instructions as a machine-readable signal. The term "machine-readable signal" refers to any signal used to provide machine instructions and / or data to a programmable processor.

[0046] The processes and logic flows described herein may be performed by one or more programmable processors, also referred to as data processing hardware, executing one or more computer programs that perform functions by manipulating input data to generate output. The processes and logic flows may also be performed by special purpose logic circuitry, such as, for example, an FPGA (field programmable gate array) or an ASIC (application specific integrated circuit). Processors suitable for executing computer programs include, by way of example, both general purpose and special purpose microprocessors, and any one or more processors of any type of digital computer. Generally, a processor receives instructions and data from a read-only memory or a random access memory, or both. The essential elements of a computer are a processor for executing instructions and one or more memory devices for storing instructions and data. Generally, a computer includes, or is operatively coupled to receive data from or transfer data to, one or more mass storage devices, such as magnetic, magneto-optical, or optical disks, for storing data. However, a computer need not include such devices. Computer-readable media suitable for storing computer program instructions and data include all forms of non-volatile memory, including, for example, semiconductor memory devices such as EPROM, EEPROM, and flash memory devices, magnetic disks such as internal hard disks and removable disks, magneto-optical disks, and CD-ROM and DVD-ROM disks. The processor and the memory can be supplemented by, or incorporated in, special purpose logic circuitry.

[0047] To provide for user interaction, one or more aspects of the present disclosure can be implemented on a computer having a display device, e.g., a CRT (cathode ray tube), LCD (liquid crystal display) monitor or touch screen, for displaying information to a user, and optionally a keyboard and pointing device, such as a mouse or trackball, by which the user can provide input to the computer. Other types of devices can also be used to provide for user interaction, e.g., feedback provided to the user can be any form of sensory feedback, e.g., visual feedback, auditory feedback, or tactile feedback, and input from the user can be acoustic, verbal, or tactile input. Additionally, the computer can interact with the user by sending and receiving documents to and from a device used by the user, e.g., by sending a web page to a web browser on the user's client device in response to a request received from the web browser.

[0048] 3A and 12-24 illustrate a schematic layout of a hollow fiber system 100 for performing automated viral vector transduction of cells. The hollow fiber system 100 includes one or more input containers 302, 302a-302d each containing an input material, one or more output containers 322, 322a-322b each configured to receive an output material, an intracapillary pump (i.e., first pump) 328, an extracapillary pump (i.e., second pump) 332, and a filter module 334 including one or more hollow fibers 336. The filter module 334 provides a convenient means for introducing various materials into the hollow fiber system 100 and retroviral materials.

[0049] FIG. 3B illustrates a horizontal cross section of a simplified example of a hollow fiber 336 of a filter module 334. The horizontal cross section is a cross section of the hollow fiber 336 along line 3B-3B shown in FIG. 3A. The hollow fiber 336 can be enclosed within a casing 337 to form an example of a filter module 334. As shown, the space within the hollow fiber 336 defines an intracapillary space 338, and the space outside the hollow fiber 336 defines an extracapillary space 339. For example, the extracapillary space 339 is the space between the hollow fiber 336 and the casing 337. While the example shown shows a single hollow fiber 336 defining the intracapillary space 338, it should be understood that there may be multiple hollow fibers 336 arranged in parallel, such as in the example shown in FIG. 3D, that cooperatively define the intracapillary space 338. That is, the filter module 334 can include any number of hollow fibers 336. One example of a filter module includes REPLIGEN's MicroKros hollow fibers.

[0050] FIG. 3C illustrates a vertical cross section of a hollow fiber 336 of the present disclosure. The vertical cross section is a cross section of the hollow fiber 136 along the line 3C-3C shown in FIG. 3A. The vertical cross section also shows the hollow fiber 336 disposed within a casing 337. The hollow fiber 336 includes a membrane having a plurality of pores that define a filter passage between an intracapillary space 338 and an extracapillary space 339. The porous membrane allows fluid to flow from the intracapillary space 338 to the extracapillary space 339 through the plurality of pores, and vice versa. As described above and shown in FIG. 3D, a plurality of hollow fibers 336 may be implemented in a filter module 334, where all of the hollow fibers 136 are housed within a casing 337. Here, each hollow fiber 336 defines a separate portion of the intracapillary space 338. With continued reference to Figures 3A and 12-24, the filter module 334 includes a pair of intra-capillary ports 360 (e.g., a first intra-capillary port 360, 360a and a second intra-capillary port 360, 360b) that are fluidly coupled to the intra-capillary space 338 at opposite ends of the filter module 334, while a pair of extra-capillary ports (e.g., a first extra-capillary port 364, 364a and a second extra-capillary port 364, 364b) are fluidly coupled to the extra-capillary space 339 at opposite ends of the filter module 334.

[0051] In some examples, the one or more input containers 302 include a transduction container 302, 302a that contains a transduction medium 304 and is fluidly coupled to a first pump 328 and a second pump 332 via a transduction conduit 340. In these examples, the one or more input containers 302 may also include a cell input container (e.g., a cell source) 302, 302b that contains a population of cells 306 and is fluidly coupled to the first pump 328 and the second pump 332 via a cell conduit 344.

[0052] The cell population 306 (hereinafter also referred to simply as "cells 306") may include immune cells or stem cells suitable for use in cell therapy, such as B cells, αβ T cells, γδ T cells, NK (natural killer) cells, iPSCs, iPSC-derived NK cells, hematopoietic stem cells, monocytes, other lymphoid cells or progenitor cells. In some embodiments, the vector includes a chimeric antigen receptor (CAR) construct that is expressed on the surface of the transduced cells after transduction using the systems and / or methods according to the present disclosure.

[0053] In some examples, once harvested, the transduced cells 306 are separated from the non-transduced cells 306 and the vector 310 by using any suitable means in the art, such as affinity isolation of the transduced cells 306 from the vector 310 and non-transduced cells 306 using an antibody that binds to the CAR expressed on the cells 306 of the transduced cells 306, or by using flow cytometry. Other suitable means in the art that can be used include, but are not limited to, size exclusion separation or some other method, such as the use of columns, membranes, etc. to separate the cells 306 from the vector 310. Once the cells 306 are isolated, separated or removed after the harvesting step, the cells 306 may be expanded and then cryopreserved, or cryopreserved after the recovery step, after which the cryopreserved cells 306 can be used for later therapeutic use.

[0054] The one or more input containers 302 may include a culture container 302, 302c that contains a culture medium 308 and is fluidly coupled to a first pump 328 and a second pump 332 via a culture conduit 346. Optionally, the hollow fiber system 100 may include an air filter 312 that is fluidly coupled to the first pump 328 and the second pump 332 via an air filter conduit 342. The one or more input containers 302 may also include a vector container (e.g., a viral or non-viral vector source) 302, 302d that contains a virus or vector 310 and is fluidly coupled to the first pump 328 and the second pump 332 via a first vector conduit 348, 348a and a second vector conduit 348, 348b.

[0055] The vector 310 may include a virus or a non-viral particle suitable for transduction of a cell. In some embodiments, the vector 310 includes a virus that includes a cargo to be delivered to the cell. In some embodiments, such cargo includes a CAR construct that is expressed on the surface of the cell after transduction with the viral vector. The virus may be derived from a lentivirus, a retrovirus, an adenovirus, an adeno-associated virus, or a hybrid virus. In some examples, the vector 310 includes a non-viral vector instead of a virus. Here, the non-viral vector may include liposomes, lipid particles, carbon, non-reactive metals, gelatin, polyamine nanospheres, and / or inorganic nanoparticles. Further examples of non-viral vectors include, for example, spheroplasts, erythrocyte ghosts, colloidal metals, inorganic nanoparticles, DEAE dextran plasmids, or combinations thereof, among others. In some embodiments, the inorganic nanoparticles are calcium phosphate nanoparticles. The vector 310 may include, among others, non-viral vectors, viral vectors, and any combination thereof.

[0056] In the illustrated examples, the first pump 328 includes a first pump inlet 327 in fluid communication with each of the input containers 302 and the air filter 312 via corresponding conduits 340, 342, 344, 346, 348. In these examples, the second pump 332 may similarly include a second pump inlet 331 in fluid communication with each of the input containers 302 and the air filter 312 via corresponding conduits 340, 342, 344, 346, 348. In some cases, it may be desirable to have only a subset of the transduction medium 304, cells 306, culture medium 308, and virus 310 (collectively referred to as input materials 304, 306, 308, 310) in fluid communication with the first pump 328 and / or the second pump 332. To that end, the hollow fiber system 100 includes a plurality of flow valves 314, 314a-314n and a plurality of pinch valves 316, 316a-316n that selectively allow fluid flow from the input container 302 to the first pump 328 and / or the second pump 332.

[0057] The flow valve 314 may include a manual pinch valve that requires user input to open or close the flow valve 314. As used herein, a flow valve 314 or pinch valve 316 in an "open" state allows fluid to flow through the respective valve, while a flow valve 314 or pinch valve 316 in a "closed" state does not allow fluid to flow through the respective valve. In some examples, the flow valve 314 is in electrical communication with the controller 200, allowing the controller 200 to direct the flow valve 314 to open or close without user input. In an open state, the flow valve 314 allows contents of the input container 302 or output container 322 to enter the hollow fiber system 100. Conversely, in a closed state, the flow valve 314 prevents contents from entering the hollow fiber system 100 from the input container 302 or output container.

[0058] The pinch valves 316 can be in electrical communication with the controller 200, allowing the controller 200 to instruct the pinch valves 316 to open and close according to a recipe, which will be described in more detail below. That is, the controller 200 can instruct each pinch valve 316 of the plurality of pinch valves 316 to open and close independently of the other pinch valves 316. Thus, by instructing the plurality of pinch valves 316 to open and close, the controller 200 can set a fluid path through which the input materials 304, 306, 308, 310 will flow. In some examples, the hollow fiber system 100 also includes a plurality of air bubble sensors 318 that detect the presence of air bubbles (e.g., air) in the input materials 304, 306, 308, 310 and help ensure that the filter module 334 receives the input materials 304, 306, 308, 310 without air bubbles. Thus, the air bubble sensors 318 are in electrical communication with the controller 200 to provide an air content value indicative of the presence of air bubbles.

[0059] In the illustrated example, each input container 302 of the plurality of input containers 302 is associated with a respective flow valve 314, pinch valve 316, and bubble sensor 318 disposed along a corresponding conduit 340, 342, 344, 346, 348 between the first pump inlet 327 and the respective input container 302. Additionally, each input container 302 of the plurality of input containers 302 is associated with a respective flow valve 314, pinch valve 316, and bubble sensor 318 disposed along a corresponding conduit 340, 342, 344, 346, 348 between the second pump inlet 331 and the respective input container 302. Specifically, the pinch valve 316 is disposed between the flow valve 314 and the bubble sensor 318 such that the pinch valve 316 is downstream of the flow valve 314 and upstream of the bubble sensor 318 with respect to the direction of flow from each input container 302.

[0060] Depending on the configuration of the flow valve 314 and / or the configuration of the pinch valve 316, a first pump inlet 327 of a first pump 328 receives one or more of the input materials 304, 306, 308, 310 and delivers the input materials 304, 306, 308, 310 at a desired rate to a pair of intra-capillary ports 360 of the filter module 334. In the illustrated example, a first pump outlet 329 of the first pump 328 is in selective fluid communication with the pair of intra-capillary ports 360. That is, the first pump outlet 329 is fluidly coupled to the first intra-capillary port 360a via the first intra-capillary conduits 356, 356a and to the second intra-capillary port 360b via the second intra-capillary conduits 356, 356b. Specifically, pinch valves 316g, 316h control the flow of fluid from the first pump outlet 329 to the first intra-capillary port 360a, and pinch valves 316i, 316j, 316k control the flow of fluid from the first pump outlet 329 to the second intra-capillary port 360b.

[0061] Continuing with the example, depending on the configuration of flow valve 314 and / or the configuration of pinch valve 316, a second pump inlet 331 of second pump 332 receives one or more of input materials 304, 306, 308, 310 and delivers them at a desired rate to a pair of extra-capillary ports 364 of filter module 334. That is, second pump outlet 333 is fluidly coupled to a first extra-capillary port 364a via a first extra-capillary sub-conduit 368, 368a and to a second extra-capillary port 364b via a second extra-capillary sub-conduit 368, 368b. Specifically, pinch valve 316m controls the flow of fluid from the second pump outlet 333 to the first extra-capillary port 364a, and pinch valves 316p, 316o control the flow of fluid from the second pump outlet to the second extra-capillary port 364b.

[0062] In some examples, the first fluid sensor 320a is disposed between the first pump outlet 329 and the first intra-capillary port 360a, and the second fluid sensor 320b is disposed between the second pump outlet 333 and the first extra-capillary port 364a. The fluid sensor 320 is configured to ensure that the first pump 328 and the second pump 332 provide accurate pressures to the first intra-capillary port 360a and the first extra-capillary port 364a, respectively. Thus, the fluid sensor 320 is in electrical communication with the controller 200 and provides the controller 200 with pressure values ​​measured at the first intra-capillary port 360a and the first extra-capillary port 364a.

[0063] In some examples, the one or more output containers 322 include a waste container 322, 322a that contains waste 324 and is fluidly coupled to the second intra-capillary port 360b and the second extra-capillary port 364b via a waste conduit 350. In these examples, the waste container 322a is also fluidly coupled to the first extra-capillary port 364a via a first extra-capillary sub-conduit 368a and a waste conduit 350. In some embodiments, the one or more output containers 322 may include a collection container 322, 322b that contains the collection medium 326 and is fluidly coupled to the second intra-capillary port 360b and the second extra-capillary port 364b via a collection conduit 352. In these embodiments, the collection container 322b is also fluidly coupled to the first extra-capillary port 364a via the first extra-capillary sub-conduit 368a and a collection conduit 352.

[0064] In the illustrated example, each output container 322 of the plurality of output containers 322 is associated with a respective flow valve 314, pinch valve 316, and air bubble sensor 318 disposed along a corresponding conduit 350, 352 between the second intra-capillary port 360b and the second extra-capillary port 364b. Specifically, the pinch valve 316 is disposed between the flow valve 314 and the air bubble sensor such that the pinch valve 316 is upstream of the flow valve 314 and downstream of the air bubble sensor 318 relative to the direction of flow into each output container 322. Continuing the example, waste container 322a is associated with pinch valves 316k, 316n disposed between waste container 322a and the second intra-capillary port 360b, and pinch valve 316o disposed between waste container 322a and the second extra-capillary port 364b. Additionally, the collection container 322b has associated therewith pinch valves 316j, 316k, 316r disposed between the waste container 322a and the second intra-capillary port 360b, and pinch valves 316j, 316n, 316o, 316r disposed between the waste container 322a and the second extra-capillary port 364b. Thus, depending on the configuration of the flow valve 314 and / or the configuration of the pinch valve 316, the output container 322 receives fluid flow from the filter module 334.

[0065] The GUI 180 is configured to receive a request from a user to operate one of the components of the hollow fiber system 100. Thus, the GUI 180 and / or the controller 200 are in operative communication with each of the components of the hollow fiber system 100. The request may include instructions to operate the first pump 328 and the second pump 332 according to a recipe that includes one or more steps. For example, the GUI 180 may instruct the first pump 328 and the second pump 332 to operate according to the recipe. In another example, the GUI 180 may instruct the plurality of pinch valves 316 to open or close according to the recipe. In yet another example, the GUI 180 is in operative communication with the fluid sensor 320 and the bubble sensor 318 such that the GUI 180 displays to the user the values ​​that the GUI 180 receives from the fluid sensor 320 and the bubble sensor 318. In some cases, the GUI 180 is configured to display the recipe to the user from the memory hardware 220 in response to receiving a request from the user.

[0066] 4 illustrates an example configuration of operations for a method 400 of operating the GUI 180 in user and developer modes. The method 400 begins at operation 402 and enters an initialization routine at operation 404. The initialization routine 404 is configured to perform basic checks of the components and connections of the hollow fiber system 100. For example, the initialization routine may determine whether the pinch valve 316, the first pump 328, and the second pump 332, the air bubble sensor 318, and the fluid sensor 320 are in a default state and / or are operational. The initialization routine may also ensure that the controller 200 and / or the GUI 180 can communicate with all components of the hollow fiber system 100. The initialization routine must be successfully completed before proceeding to operation 412 (i.e., user mode) or operation 422 (i.e., developer mode).

[0067] Upon successful completion of the initialization routine 404, the method 400 may proceed to either operation 412 or operation 422. That is, the GUI 180 may receive a user input requesting that the hollow fiber system 100 be operated in a user mode and proceed to operation 412. In the user mode, the user may select one or more recipes to be executed by the hollow fiber system 100. Once the user selects a recipe, the method 400 proceeds to operation 414 to execute the selected recipe. Recipe execution is described in more detail below, but in the user mode, the GUI 180 restricts the user to only execute recipes that are stored in the memory hardware 220. That is, in the user mode, the user may be restricted to execute recipes from a number of predefined recipes stored in the memory hardware 220. The GUI 180 may display information about the recipe to the user while the recipe is being executed. Once the recipe execution is complete, the method 400 ends in operation 430.

[0068] Alternatively, after the initialization routine 404, the GUI 180 may receive a user input requesting to operate the hollow fiber system 100 in a developer mode and proceed to operation 422. When in developer mode, the user may create or execute the method 400, and proceed to operation 424 to allow the user to create and / or execute recipes. Thus, in developer mode, the GUI 180 may prompt the user to enter credentials that prove the user has permission to operate the GUI in developer mode. After creating a new recipe or executing a recipe, the method ends at operation 430.

[0069] Returning to Figures 3A and 12-24, in some embodiments, the hollow fiber system 100 includes a mixer 195 configured to mix one or more of the input materials 304, 306, 308, 310 prior to loading the materials into the filter module 334. In some cases, pre-mixing the input materials increases the interaction of the one or more of the input materials 304, 306, 308, 310 during transduction. For example, the mixer 195 can mix cells 306 and viruses (i.e., vectors) 310 to generate a mixture thereof and then load the cells 306, viruses 310, or mixture into the respective input containers 302. In these examples, pre-mixing the cells 306 and viruses 310 prior to transduction increases the interaction (e.g., collisions) between the cells 306 and viruses 310 during transduction. In the illustrated example, the mixer 195 is shown as a peripheral device operating in parallel with the hollow fiber system 100, allowing the inputs to be pre-mixed before being fed into one of the input containers. Alternatively, the hollow fiber system 100 may include one or more mixers 195 integrated between the input container 302 and one or both of the pumps 328, 332, allowing the input materials 304, 306, 308, 310 to be mixed together before being distributed to the filter module 334. Optionally, the mixer 195 may include a conduit that fluidly couples the mixer 195 to one or more conduits or one of the input containers 302 of the hollow fiber system 100.

[0070] In some embodiments, the hollow fiber system 100 includes a welder 190 configured to weld the input container 302 or the output container 322 to the corresponding conduit 340, 342, 344, 346, 348, 350, 352. That is, one or more of the input containers 302 and one or more of the output containers 322 can be removed from or attached to the hollow fiber system 100 by the welder 190. For example, when loading the transduction container 302a into the hollow fiber system 100, the welder 190 can weld the transduction container 302a to the transduction conduit 340. Thus, the welder 190 provides a sealed connection between the transduction container 302a and the hollow fiber system 100.

[0071] Embodiments herein are further directed to computer-implemented methods that, when executed on the data processing hardware 210 of the controller 200, cause the data processing hardware 210 to perform operations on the hollow fiber system 100. In particular, the controller 200 may store multiple recipes 500 in the memory hardware 220. With reference to FIG. 5A, each recipe 500 includes multiple steps 510, 510a-510n, where each step 510 of the recipe 500 instructs the hollow fiber system 100 to perform an operation. In some embodiments, the recipe 500 is a CSV file or other data table, where each row of data corresponds to a step 510 of the recipe 500 and each column of data corresponds to a component instruction (e.g., valve configuration, pump speed, etc.) for the respective step. Thus, the controller 200 repeats each step 510 of the recipe 500 until the last step of the recipe 500 has been executed. In particular, recipe 500 automates the control of hollow fiber system 100 , thereby allowing a user to perform cell transduction with little or no interaction with hollow fiber system 100 .

[0072] In the illustrated example, each step 510 of the recipe 500 includes at least one of a valve configuration state 512, a pump direction state 514, a pump speed state 516, a step type 518, and an end trigger threshold 520. Thus, in each step 510 of the recipe 500, the controller 200 instructs the hollow fiber system 100 (e.g., via one or more command signals 212, 214, 216, 218, 221, 231, as shown in FIG. 1D ) according to at least one of the valve configuration state 512, the pump direction state 514, the pump speed state 516, the step type 518, and the end trigger threshold 520. Thus, in some examples, the controller 200 can instruct the hollow fiber system 100 according to the valve configuration state 512 and the pump speed 516 in a single step of the recipe 500. The valve configuration state 512 can instruct the controller 200, via the flow valve command signal 214 and the pinch valve command signal 216 (FIG. 1D), to open or close the flow valves 314 and / or the pinch valves 316, respectively. In some examples, the valve configuration state 512 instructs the controller 200 to open all the pinch valves 316 or to close all the pinch valves 316. In other examples, the valve configuration state 512 instructs the controller 200 to instruct ... Controller 200 can receive flow valve signal 213 and pinch valve command signal 216 (FIG. 1D) to determine whether each flow valve 314 and each pinch valve 316, respectively, is open or closed.

[0073] The pump direction state 514 directs the fluid flow direction of the pumps 328, 332 via the pump command signal 231 (FIG. 1D). For example, the pump direction state 514 can specify whether fluid flows in a first direction (e.g., toward the output container 322) or a second direction (e.g., toward the input container 302). In some examples, the pump direction state 514 directs the controller 200 to operate the first pump 328 in a first direction and the second pump 332 in a second direction. In other examples, the pump direction state 514 directs the controller to operate both the first pump 328 and the second pump 332 in the first direction or the second direction. In some implementations, the pump direction state 514 alternates for each step 510 of the recipe 500. For example, in a first step 510a of the recipe 500, the pumps 328, 332 may operate in a first direction, and in a second step 510b of the recipe, the pumps 328, 332 may change fluid flow direction and operate in a second direction. The controller 200 may determine a current pump direction state 514 of the first pump 328 and the second pump 332 based on receiving pump signals 229 from the pumps 328, 332 (FIG. 1D).

[0074] The pump speed 516 instructs the controller 200, via the pump command signal 231 (FIG. 1D), to operate the pumps 328, 332 at a specified speed, such as revolutions per minute (RPM). The pump speed 516 may directly correlate to the fluid flow rate or pressure that the pumps 328, 332 generate. That is, operating the pumps 328, 332 faster increases the fluid flow rate or pressure, and operating the pumps 328, 332 slower decreases the fluid flow rate or pressure. Thus, the controller may determine the pump speed 516 based on a target pressure or fluid flow parameter. In some examples, a pump speed 516 of "0" indicates that the pumps 328, 332 are not currently pumping fluid. In these examples, the first pump 328 may be pumping fluid while the second pump 332 is not pumping fluid, or vice versa. Based on receiving pump signals 229 from the pumps 328, 332, the controller 200 can determine the current pump speed state 516 of the first pump 328 and the second pump 332 (FIG. 1D).

[0075] The step type 518 specifies the characteristics of the termination trigger threshold 520 of each step 510 of the recipe 500. That is, the characteristics of the step type 518 represent the conditions that must be met for the step 510 to complete. For example, the step type 518 may include characteristics of fluid detection, timer, air detection, fluid vs. air detection, air vs. fluid detection, or pressure detection. That is, the fluid detection and air detection step types 518 specify that the step 510 of the recipe 500 runs until a particular fluid detection or air detection parameter, respectively (e.g., a target fluid detection parameter or a target air detection parameter) is met. The timer specifies that the step 510 runs for a predetermined amount of time.

[0076] FIG. 5B illustrates an example of a termination trigger threshold 520 for step 510 of recipe 500. The termination trigger threshold 520 specifies a particular condition that, if met, completes execution of step 510 or terminates execution of step 510 and / or recipe 500. In some examples, one or more steps 510 of recipe 500 include multiple termination trigger thresholds 520. Specifically, a first termination trigger threshold 520 may indicate a particular operating condition that, if met, completes execution of step 510. That is, when hollow fiber system 100 meets the condition of the first termination trigger threshold 520 of step 510, controller 200 successfully completes execution of step 510. Controller 200 may use one or more received signals 211, 213, 215, 217, 219, 229 (see FIG. 1D) to determine whether hollow fiber system 100 meets the condition of the termination trigger threshold 520. For example, step 510 may include a first termination trigger threshold 520 for generating a pressure of 15 PSI, where the first termination trigger threshold 520 represents a target value that step 510 must achieve in order for hollow fiber system 100 to operate properly. In this example, once hollow fiber system 100 meets the condition of first termination trigger threshold 520 (e.g., meets 15 PSI pressure), controller 200 successfully completes execution of step 510 and proceeds to the next step 510 of recipe 500.

[0077] Continuing with the above example, step 510 of recipe 500 may include a second termination trigger threshold 520 indicating a particular, different condition that, if met, will terminate execution of step 510. As such, if hollow fiber system 100 exceeds the condition of second termination trigger threshold 520 of step 510, controller 200 will terminate execution of step 510 and / or the entire recipe 500. For example, step 510 may include a second termination trigger threshold 520 that prevents pressure from exceeding 50 PSI (e.g., burst pressure). Here, second termination trigger threshold 520 represents a fail-safe value that hollow fiber system 100 should not exceed. In this example, if hollow fiber system 100 exceeds second termination trigger threshold 520 (e.g., pressure exceeds 50 PSI), controller 200 will terminate execution of step 510 or terminate execution of the entire recipe 500. In some examples, the second termination trigger threshold 520 indicates a minimum value that the hollow fiber system 100 should not fall below (e.g., a minimum of 10 PSI). In yet other examples, step 510 of recipe 500 includes the second termination trigger threshold 520, as described above. Alternatively, the second termination trigger threshold 520 is included in a process that runs in the background while the hollow fiber system 100 is operating.

[0078] In response to the hollow fiber system satisfying the second termination trigger threshold 520, the controller 200 can terminate the recipe 500, terminate step 510, initiate a pump rate adjustment procedure, and / or prompt the user via the GUI 180. Terminating the recipe 500 or step 510 may be undesirable since it may stall the cell transduction process. Thus, the controller 200 can execute a pump rate adjustment procedure when the hollow fiber system 100 is within a threshold (e.g., 10%) of the second termination trigger threshold 520. The pump rate adjustment procedure is described in more detail with reference to FIG. 8. In particular, by initiating the pump rate adjustment procedure, the controller 200 prevents the hollow fiber system 100 from satisfying the second termination trigger threshold 520, allowing the recipe 500 to continue to run without any errors. In some cases, the controller 200 prompts the user of the hollow fiber system 100 via the GUI 180. The prompts may allow the user to provide user input in the GUI 180 to instruct the controller 200 to skip (i.e., ignore) step 510, retry execution of step 510, modify the termination trigger threshold 520, and / or allow the user to provide additional user input to complete execution of the recipe 500.

[0079] In some embodiments, the recipe steps 510 include a first termination trigger threshold 520 that indicates to the controller 200 what conditions must be met to complete the respective step 510, and a second termination trigger threshold 520 that indicates to the controller 200 what conditions must be met to terminate the step 510. For example, the first termination trigger threshold 520 indicates a target pressure of 25 PSI and the second termination trigger threshold 520 indicates a minimum pressure of 5 PSI. In this example, once the hollow fiber system 100 reaches the target pressure of 25 PSI, the controller 200 successfully completes execution of the step 510. Continuing the example, once the hollow fiber system 100 falls below the minimum pressure of 5 PSI, the controller 200 terminates execution of the step 510. In another example, the first termination trigger threshold 520 may indicate a target pump run time of 5 minutes and the second termination trigger threshold indicates a maximum pressure of 50 PSI. Now, once the pumps 328, 332 have been running for 5 minutes, the controller 200 successfully completes the execution of step 510. In parallel, the controller 200 also monitors the pressure in the hollow fiber system 100, and once the pressure exceeds 50 PSI, the controller 200 terminates the execution of step 510.

[0080] The termination trigger threshold 520 indicates at least one of a termination trigger type 522, a pressure value 524, a stopwatch timer value 526, an air volume value 528 (e.g., air bubble detection), or a fluid flow value (e.g., fluid flow rate, direction, or other fluid flow parameter) 530. Here, the controller 200 determines the pressure value 524 based on a fluid flow signal 219 (FIG. 1D) received from one of the fluid sensors 320. Additionally, the controller 200 determines the air volume value 528 using an air bubble sensor signal 217 (FIG. 1d) received from one of the air bubble sensors 318. In some examples, the controller 200 determines the fluid flow value 530 from the fluid flow signal 219 (FIG. 1D) received from one of the fluid sensors 320 (e.g., a fluid flow sensor). In other examples, the controller 200 determines the fluid flow value 530 using a pump speed 516 of the pump 328, 332. In yet another example, controller 200 uses a combination of pump speed 516 and fluid flow signal 219 (FIG. 1D) to determine fluid flow value 530. Optionally, termination trigger threshold 520 includes a sensor identification that identifies which sensor is associated with termination trigger threshold 520 (e.g., which sensor to obtain a signal from). For example, termination trigger threshold 520 illustrated in FIG. 5B identifies one of air bubble sensors 318 of multiple air bubble sensors 318 to read air volume value 528 and / or one of fluid sensors 320 to read pressure value 524.

[0081] The termination trigger type 522 can specify the type of termination trigger threshold 520. The termination trigger types 522 include timer, pressure rise, pressure drop, fluid to air ratio, or integrity. The integrity termination trigger type 522 refers to a pressure hold of the hollow fiber system 100, where the hollow fiber system 100 holds the pressure at a specified value and measures the pressure drop over a period of time, thereby indicating the integrity of the hollow fiber system (e.g., whether a leak is present), as will be described in more detail with reference to FIG. 16. The termination trigger type 522 can instruct the controller 200 to obtain a particular value from the hollow fiber system 100. The pressure value 524 specifies a minimum pressure value and / or a maximum pressure value. For example, the pressure value 524 specifies a maximum value of 30 pounds per square inch (PSI) and a minimum value of 10 PSI.

[0082] The timer value 526 may specify a maximum or minimum execution time for the step 510 of the recipe 500. For example, a step 510 with an average execution time of 10 seconds may have a timer value 526 (e.g., a watchdog timeout value) of 60 seconds. Thus, if the execution time of the step 510 exceeds 60 seconds, the controller 200 may determine that there is an error in the execution of the step 510. For example, the first termination trigger threshold 520 may indicate a target pressure of the hollow fiber system 100, and the second termination trigger threshold 520 indicates a watchdog timer value 526 (e.g., 1 minute). Here, the watchdog timer value 526 indicates that if the hollow fiber system 100 does not achieve the target pressure of the first termination trigger threshold 520 within 1 minute, an error must have occurred in the hollow fiber system 100. That is, the error may include, but is not limited to, a hole in the hollow fiber membrane, a hole in the conduit, and / or overfilling (or underfilling) of the input container 302. In this example, the timer value 526 indicates to the controller 200 when to finish executing the step. In some implementations, the timer value 526 indicates a target time for completing the execution of the step. For example, the timer value 526 for step 510 may include a run time of the first pump 328 for 5 minutes, such that after the first pump 328 has been running for 5 minutes, the controller 200 has successfully completed execution of step 510 and proceeded to the next step 510.

[0083] The air content value (e.g., air value) 528 specifies the amount of air that is acceptable in the fluid (i.e., input material). That is, the air bubble sensor 318 provides the air content value to the controller 200 to ensure that the filter module 334 receives the input material 304, 306, 308, 310 without air bubbles. The fluid flow value 530 specifies a maximum or minimum amount of fluid flow. The fluid flow value 530 may represent an amount of fluid per unit time, such as, for example, liters per minute. In some examples, the air bubble sensor 318 provides the fluid flow value 530 to the controller 200. In other examples, the fluid sensor 320 provides the fluid flow value 530 to the controller 200 (e.g., via the fluid flow signal 219).

[0084] 5B, the termination trigger threshold 520 may include a fluid sensor 320 identifier, a termination trigger type 522, a pressure value 524, and a timer value 526 (each shown as a solid line), but does not include an air value 528 and a fluid flow value (each shown as a dotted line). In this example, the termination trigger threshold 520 specifies a first fluid sensor 320, a pressure rise termination trigger type 522, a maximum pressure value 524, and a timer value 526, where each value includes a logical "OR" or "AND" operation such that only one of the values ​​needs to be met to satisfy the termination trigger threshold 520 and all of the values ​​need to be met to satisfy the termination trigger threshold 520 (e.g., to complete or terminate execution of step 510). Continuing with the example, the watchdog timeout value 526 indicates a target execution time of 2 minutes for the pumps 328, 332 to successfully complete step 510, and the maximum pressure value 524 indicates whether execution of step 510 and / or the recipe 500 will terminate if one of the fluid sensors 320 exceeds 50 PSI, where the controller 200 will either complete execution or terminate execution of step 510 if either condition is met.

[0085] In another example, the timer value 526 may indicate a target run time of 2 minutes for the pumps 328, 332 and the pressure value indicates a target pressure of 25 PSI to complete execution of step 510. In this alternative example, the logic operation may require both conditions to be met before the controller 200 completes execution of the step. However, in some cases, the logic operation may require only one of the conditions to be met for the controller 200 to complete execution of step 510. In some examples, the termination trigger threshold 520 specifies a timed termination trigger type 522 and timer value 526 at the first fluid sensor 320. In other examples, the termination trigger threshold 520 specifies a fluid vs. air termination trigger type 522 and timer value 526 at the third air bubble sensor 318. Thus, the termination trigger threshold 520 defines operating parameters that define acceptable and / or target operating conditions for the hollow fiber system. As used herein, operating parameters may collectively refer to at least one of termination trigger type 522, pressure value 524, timer value 526, air value 528, fluid flow value 530, and / or any signals received by controller 200 from the hollow fiber system (e.g., signals 211, 213, 215, 217, 219, 229 shown in FIG. 1D).

[0086] In some scenarios, a user of the hollow fiber system 100 may create and save a recipe 500 to the memory hardware 220 for later execution. While the system is in developer mode, the user creates the recipe 500 via the GUI 180. The memory hardware 220 communicates with the data processing hardware 210 to enable the data processing hardware 210 to execute one of the stored recipes 500. The user may create the recipe 500 by providing user input to the GUI 180 that specifies parameters for each step of the recipe 500. The GUI 180 may then receive a request to execute the recipe 500 stored in the memory hardware 220. In response to the request, the controller 200 may retrieve the recipe 500 stored in the memory hardware 220 and execute the recipe 500 on the hollow fiber system 100 using the data processing hardware 210.

[0087] In some implementations, the GUI 180 receives a request to modify at least one step of a recipe stored in the memory hardware 220. The modification request may specify at least one parameter to change for a step 510 of the recipe 500. For example, a user input to the GUI 180 may specify that a plurality of pinch valves 316 are to be changed from an open state to a closed state for a first step 510 of the recipe 500. In another example, a user input to the GUI 180 modifies a timer value 526 from 30 seconds to 60 seconds for the step 510 of the recipe. After receiving the modification request, the controller 200 updates at least one step of the recipe 500 stored in the memory hardware 220.

[0088] 6 illustrates an example configuration of operations of the method 600 for executing an initialization routine via the GUI 180. A user may begin the method 600 at operation 602 by loading an input container 302 or an output container 322 into the hollow fiber system 100. Loading the container 302, 322 refers to fluidly connecting the container 302, 322 to the hollow fiber system 100. For example, the welder 190 may load the container 302, 322 by welding the container 302, 322 to a conduit of the hollow fiber system 100. At operation 604, the method 600 includes executing an initialization routine at the GUI 180 to check communication and / or electrical connections of the components of the hollow fiber system 100. The initialization routine may be executed in response to a user input at the GUI 180 or in response to a decision by the controller 200 to execute the recipe 500. If, at operation 606, no control hardware (e.g., controller 200) is found during the initialization routine, method 600 proceeds to operation 608 and displays an error to the user via GUI 180. After displaying the error, method 600 ends at operation 610. Alternatively, if control hardware is found during the initialization routine, method 600 proceeds to operation 612 and opens all valves (e.g., pinch valve 316 and flow valve 314). With all valves open, at operation 614, method 600 includes determining whether a tube set 160 (FIG. 1C) is loaded into hollow fiber system 100. Alternatively, method 600 may determine whether an input container 302 or an output container 322 is loaded into hollow fiber system 100. If tube set 160 is not inserted, method 600 waits until tube set 160 is inserted. Alternatively, once the tubeset 160 is inserted, the method 600 includes closing all valves at operation 616. The controller may close all flow valves 314 and / or all pinch valves 316. After closing all valves, at operation 618, the GUI 180 may receive user input to operate in a user mode at operation 620 or user input to operate in a developer mode at operation 622.

[0089] FIG. 7 illustrates an example configuration of operations of method 700 for executing each step 510 of recipe 500. Here, data processing hardware 210 of controller 200 may execute each operation of method 700. In operation 702, method 700 reads recipe 500 stored in memory hardware 220 of controller 200. Here, recipe 500 may include a data file including lines corresponding to each step 510 of recipe 500. In operation 704, method 700 sets a line counter to zero, allowing controller 200 to begin execution at the first step 510 of recipe 500. In operation 708, method 700 determines whether welder 190 has welded the required ports. In some examples, a user provides user input via GUI 180 to indicate whether the required ports have been welded. Alternatively, controller 200 may determine whether the required ports have been welded. If the required ports have not been welded, method 700 proceeds to operation 706 and waits for an acknowledgement of the port welding before proceeding to operation 710. If the required ports are welded, then in operation 710 the method 700 directs the plurality of pinch valves 316 to transition to an open or closed state according to the valve configuration state 512 of the current step 510 of the recipe 500. The controller 200 may provide instructions to the plurality of pinch valves 316 via the pinch valve command signals 216. Then, in operation 712, the method 700 directs the pumps 328, 332 to operate according to the pump direction state 514 and / or pump speed 516 of step 510.

[0090] In operation 714, the method 700 determines whether the execution time of the currently executing step 510 of the recipe 500 exceeds the timer value 526 (e.g., a watchdog timer value) for that step. If the execution time of the currently executing step 510 exceeds the timer value 526, then in operation 716, the method 700 stops the pumps 328, 332 (e.g., sets the pump speed 516 to zero via the pump command signal 231) and prompts the user via the GUI 180 to retry, skip, or abort the step 510. If the user provides user input to abort the currently executing step 510, then the method 700 terminates execution of the entire recipe 500 in operation 728. Alternatively, if the user provides user input to retry the currently executing step 510, then the method 700 decrements the row counter in operation 718 and proceeds to operation 726. Specifically, decrementing the row counter allows the controller 200 to return to the previous step 510 in the recipe 500 (or the beginning of the currently executing step 510) and retry the step 510 where the watchdog fault occurred. If the user provides user input to skip the currently executing step 510, the method 700 increments the row counter in operation 724 and skips the currently executing step 510 even if the currently executing step 510 has not completed execution successfully.

[0091] In the illustrated example, if the execution time of step 510 does not exceed the timer value 526, then the method 700 determines whether the termination trigger threshold 520 of step 510 is met at operation 720. Here, if the termination trigger threshold 520 is not met, the method 700 returns to operation 714. If the termination trigger threshold 520 is met, then at operation 722, the method 700 initiates a pump rate adjustment procedure. Here, the pump rate adjustment procedure includes stopping operation of the pumps 328, 332 for a predetermined period of time. Alternatively, the pump rate adjustment procedure includes reversing the pump direction 514 of the pumps 328, 332. Thereafter, at operation 724, the method 700 increments a row counter (e.g., transitions to the next step 510 of the recipe 500).

[0092] At operation 726, the method 700 determines whether the currently executing step 510 of the recipe 500 is the last step of the recipe 500. If the currently executing step 510 of the recipe 500 is the last step 510 of the recipe 500, then at operation 728, the method 700 completes the execution of the recipe 500. If the currently executing step 510 of the recipe 500 is not the last step 510 of the recipe 500, the method 700 returns to operation 706. Operations 702-726 are repeated for each step 510 of the recipe 500 until the last step 510 of the recipe 500 is completed.

[0093] In some implementations, the controller 200 executes the recipe 500 that includes pressure-based control. Pressure-based control includes performing an action in response to the controller 200 determining whether the pressure value 524 exceeds a maximum threshold or does not exceed a minimum threshold. For example, the controller 200 may instruct the first pump 328 (via the pump command signal 231) to operate at a first pump speed 516 to provide a first flow of at least one of the input materials 304, 306, 308, 310 to the first intra-capillary port 360a according to a first step 510 of the recipe 500. Alternatively, the controller 200 may instruct the second pump 332 to operate at a first pump speed 516 to provide a first flow of at least one of the input materials 304, 306, 310 to the first extra-capillary port 364a. For example, the first pump 328 can supply cells 306 and / or vectors 310 to the first intracapillary port 360a. In another example, the first pump 328 can supply culture medium 308 and / or transduction medium 304 to the first intracapillary port 360a. Here, in a first step 510 of the recipe 500, the controller 200 obtains a first fluid flow parameter 530 of the fluid flow of at least one input substance 304, 306, 308, 310 from the fluid sensor 320a. In this example, the first step 510 of the recipe 500 can include a termination trigger threshold 520 based on a maximum pressure value 524 to prevent damage to components of the hollow fiber system 100 (e.g., bursting of a conduit). Thus, when the pressure value 524 of the first flow exceeds the maximum pressure threshold, the controller 200 can initiate a pump rate adjustment procedure.

[0094] 8 illustrates an example configuration of operations of a method 800 for performing a pump rate adjustment procedure. In operation 802, the method 800 determines whether the termination trigger threshold 520 of the respective step 510 has been met. In some examples, the controller 200 determines whether the hollow fiber system 100 is within a threshold (e.g., 10%) of the termination trigger threshold 520. If the termination trigger threshold 520 is not met, in operation 804, the controller 200 determines whether the first measured fluid flow parameter 530 from the fluid sensor 320 is within a threshold (e.g., 10%) of the termination trigger threshold 520. M1 The controller 200 obtains the first measured fluid flow parameter 530 via the first fluid flow signal 219. M1 Further, the controller 200 obtains the first measured fluid flow parameter 530. M1 a first fluid flow parameter threshold 530 T1 For example, the first fluid flow parameter threshold 530 T1 may be a predetermined maximum pressure value associated with a component of the system, for example 28 pounds per square inch (PSI). M1 is the first fluid flow parameter threshold 530 T1 If not, the method 800 returns to operation 802. M is the first fluid flow parameter threshold 530 T1 If so, then at operation 806, the method 800 initiates a pump rate adjustment procedure by operating the pumps 328, 332 in a reverse flow condition (e.g., by reversing the pump direction state 514). Operating the pumps 328, 332 in a reverse flow condition is configured to reverse the direction of fluid flow and decrease the first fluid flow parameter 530. Additionally or alternatively, the pump rate adjustment procedure may include ceasing operation of the pumps 328, 332 for a predetermined period (e.g., a period of time). In some examples, the predetermined period of time is a pressure adjustment period.

[0095] In operation 808, the method 800 includes receiving a second measured fluid flow parameter 530 of the first flow from the fluid sensor 320. M2The controller 200 obtains the second measured fluid flow parameter 530 via the second fluid flow signal 219. M2 Here, the controller 200 obtains the second measured fluid flow parameter 530 a predetermined time after reversing the pumps 328, 332 in operation 806. M2 The controller 200 obtains a second measured fluid flow parameter 530 M2 the second fluid flow parameter threshold 530 T2 . The second fluid flow parameter threshold 530 T2 In particular, the second fluid flow parameter threshold 530 may be 15 PSI. T2 is the first fluid flow parameter threshold 530 T1 The second measured fluid flow parameter 530 is less than M2 is the second fluid flow parameter threshold 530 T2 If so, the method 800 obtains updated values ​​and updates the second measured fluid flow parameter 530 until the threshold is met. M2 the second fluid flow parameter threshold 530 T2 The second measured fluid flow parameter 530 is compared to M2 is the second fluid flow parameter threshold 530 T2 If not, then in operation 810, the method 800 initiates a pump rate adjustment procedure responsive to the comparison. Specifically, the pump rate adjustment procedure reduces the speed of the pumps 328, 332 by 50 percent (e.g., operates the pumps 328, 332 at the second pump speed 516 responsive to the comparison).

[0096] In operation 812, the method 800 reduces the pump speed (i.e., the measured pump speed) 516 of the pumps 328, 332 after reducing the pump speed 516 in operation 810. M Additionally, the controller 200 obtains the measured pump speed 516 M The pump speed threshold 516 T Here, the controller 200 compares the measured pump speed 516 M Pump speed threshold 516 TDetermine whether the measured pump speed is 516. M Pump speed threshold 516 T If so, then in operation 814, the method 800 restarts the pumps 328, 332. Alternatively, the measured pump speed 516 M Pump speed threshold 516 T If not, then in operation 816 the method 800 prompts the user via the GUI 180 to skip or abort the currently executing step 510 of the recipe 500. If the user chooses to skip the currently executing step 510 of the recipe 500, then in operation 820 the method 800 proceeds to the next step 510 of the recipe 500. If the user chooses to abort the currently executing step 510 of the recipe 500, then in operation 818 the method 800 instructs the pump 328 to be idle.

[0097] FIG. 9 illustrates an example configuration of operations of a method 900 for executing an example recipe 500 in a hollow fiber system 100. Here, the controller 200 can execute each operation of the method 900. In operation 902, the method 900 includes the controller 200 instructing one or more pumps 328, 332 to execute a first step of the recipe to provide a first pressure at a first fluid sensor 320. Here, the first pressure at the first fluid sensor 320 may be specified by a first step 510 of the recipe 500. Further, the controller 200 can send a pump command signal 231 to the one or more pumps 328, 332 to operate at a first pump speed 516 specified by the recipe 500. In this example, the controller 200 can also send a pump command signal 231 to the one or more pumps 328, 332 further indicating a direction of operation of the pump (e.g., according to a pump direction 514 of the first step 510 of the recipe 500) to operate in a first direction. Additionally, the controller 200 can direct the pinch valves 316 to open or close according to the valve configuration 512 of the first step 510 of the recipe 500 .

[0098] At operation 904, the method 900 includes receiving a fluid flow signal 219 (e.g., a pressure signal) indicative of an operating parameter measured at the first fluid sensor, where the operating parameter may include a pressure value 524 and / or a fluid flow value 530 at the first fluid sensor 320. At operation 906, the method 900 includes the controller 200 comparing a first termination trigger threshold 520 for the first step 510 of the recipe 500 to the fluid flow signal 219 (e.g., a pressure signal) received from the first fluid sensor 320. That is, the first step 510 of the recipe may specify a maximum pressure value 524 for the fluid pressure at the first fluid sensor 320. The maximum pressure value 524 may represent a target pressure, such that once the pressure reaches the maximum pressure value, the controller 200 has successfully executed the first step 510, and the controller 200 may complete the first step 510 of the recipe 500 and begin executing the second step 510 of the recipe 500. Alternatively, the maximum pressure value 524 may represent a fail-safe pressure. Here, when the pressure reaches the maximum pressure value 524, the controller 200 terminates the first step 510 of the recipe 500. That is, the pressure value 524 may reach a pressure that may damage the hollow fiber system 100, so the controller 200 terminates the execution of the first step 510 to protect the hollow fiber system 100. Alternatively, the controller 200 may ignore the first step 510 or prompt the user via the GUI 180. Thus, in operation 908, the method 900 includes the controller 200 instructing one of the pumps 328, 332 to complete the first step 510 of the recipe 500 based on a comparison of the received pressure signal to the first termination trigger threshold 520. In some cases, when the controller 200 terminates the first step 510 of the recipe, the controller 200 further skips or retries the first step 510 of the recipe.

[0099] 10 illustrates an example of a configuration of operations of a method 1000 for executing an example recipe 500 in a hollow fiber system 100. Here, the controller 200 can execute each operation of the method 1000. In operation 1002, the method 1000 includes the controller 200 instructing one or more pumps 328, 332 to execute a first step of the recipe 500 at a first pump speed 516 according to a first step 510 of the recipe 500. The controller 200 can determine the first pump speed 516 using a first target pressure specified in the recipe 500. In this example, the controller 200 can also send a pump command signal 231 to one or more pumps 328, 332 indicating a direction of operation of the pump (e.g., according to the pump direction 514 of the first step 510 of the recipe 500) to operate in the first direction. Additionally, the controller 200 can instruct the pinch valves 316 to open or close according to the valve configuration 512 of the first step 510 of the recipe 500 to set the fluid flow path of one or more pumps 328, 332.

[0100] At operation 1004, the method 1000 includes receiving a fluid flow signal 219 (e.g., a pressure signal) indicative of a pressure value 524 measured at a first fluid sensor 320 of the controller 200. Here, the pressure value 524 is an operating parameter. At operation 1006, the method 1000 includes the controller 200 comparing a first termination trigger threshold 520 for a first step 510 of the recipe 500 to the pressure signal received from the first fluid sensor 320. Here, the first step 510 of the recipe specifies a maximum pressure value 524 for the fluid pressure at the first fluid sensor 320. The maximum pressure may represent a burst pressure value of the filter module 334 or the conduit. Thus, at operation 1008, the method 1000 includes the controller 200 initiating a pump rate adjustment procedure based on the comparison that the received pressure signal exceeds the first termination trigger threshold 520.

[0101] The pump rate adjustment procedure may include the controller 200 operating one or more pumps 328, 332 in a reverse flow condition (e.g., changing the pump direction 514) for a predetermined period of time. In some examples, the pump direction 514 switches between a first direction and a second direction at each step 510 of the recipe 500. Alternatively, the pump rate adjustment procedure may include the controller 200 stopping operation of one or more pumps 328, 332 (e.g., operating at zero pump speed 516) for a predetermined period of time.

[0102] In particular, both redirecting and deactivating one or more pumps 328, 332 reduces the pressure at the first fluid sensor 320 such that the pressure measured at the first pressure sensor does not meet the first termination trigger threshold 520 (i.e., the measured pressure is within an acceptable range). Accordingly, during execution of the first step 510 of the recipe 500, the received pressure signal may exceed the first termination trigger threshold 520 and the controller 200 may initiate a pump rate adjustment procedure, which allows the controller 200 to continue execution of the first step 510 without terminating the first step 510. Additionally, the controller 200 initiates the pump rate adjustment procedure (e.g., without user input) in response to comparing the received pressure signal to the first termination trigger threshold 520, resulting in reduced user interaction with the hollow fiber system 100 to perform the transduction of a cell therapy.

[0103] 11 illustrates an example of a configuration of operations of a method 1100 for executing an example recipe 500 in a hollow fiber system 100. Here, the controller 200 can execute each of the operations of the method 1000. In operation 1102, the method 1000 includes the controller 200 instructing one or more pumps 328, 332 to execute a first step of the recipe 500 to operate with a first fluid flow parameter 530 (e.g., a constant fluid flow parameter) at the fluid sensor 320 (or the air bubble sensor 318). In this example, the controller 200 can also send a pump command signal 331 to the one or more pumps 328, 332 indicating a direction of operation of the pump (e.g., according to the pump direction 514 of the first step 510 of the recipe 500) to operate in the first direction. Additionally, the controller 200 can instruct the pinch valves 316 to open or close according to the valve configuration 512 of the first step 510 of the recipe 500 to set the fluid flow path of one or more pumps 328, 332.

[0104] At operation 1104, the method 1100 includes receiving a fluid flow value 219 (e.g., a pressure signal) indicative of a fluid pressure (e.g., a pressure value 524) at the first fluid sensor 320, where the pressure value is a measured operating parameter. At operation 1106, the method 1100 includes the controller 200 comparing a first termination trigger threshold 520 for the first step 510 of the recipe 500 to the pressure signal received from the first fluid sensor 320. That is, the first step 510 of the recipe may specify a maximum pressure value 524 for the fluid pressure at the first fluid sensor 320, where the maximum pressure value 524 represents a fail-safe pressure at which the controller 200 will reduce the fluid pressure at the first fluid sensor 320. At operation 1108, the method 1100 includes the controller 200 initiating a pump rate adjustment procedure based on a comparison of the received pressure signal to the first termination trigger threshold 520. That is, the controller 200 can instruct the pumps 328, 332 to operate at a second pump speed (e.g., a speed lower than the first pump speed), stop operation of the pumps 328, 332, and / or reduce the fluid flow direction of the pumps 328, 332. In particular, the controller 200 can operate the pumps 328, 332 at a constant fluid flow parameter 530 (without exceeding a maximum pressure) such that the controller 200 completes the recipe 500 in the shortest time. In particular, the controller 200 can determine a maximum fluid flow parameter 530 for operating the hollow fiber system 100 without exceeding a maximum pressure. Thus, the controller 200 can operate the pumps 328, 332 at a maximum speed to minimize the transduction time in the hollow fiber system 100.

[0105] 12-24 illustrate exemplary schematic diagrams of a fluid flow path through hollow fiber system 100 according to the steps 510 of recipe 500. Specifically, each of FIGS. 12-24 illustrates one step 510 of the steps 510 of 500. It should be understood that the steps 510 shown in FIGS. 12-24 are merely illustrative examples of recipe 500 and are not intended to limit the disclosure in any way. For example, a different example recipe 500 may repeat any of the steps 510 any number of times, may include more or fewer steps 510, and / or may perform the steps 510 in a different order than the example recipe 500 illustrated in FIGS. 12-24. For example, a recipe 500 may include only one step 510 of the example recipe 500 illustrated in FIGS. 12-24. In other examples, the recipe 500 can include one or more additional steps 510 not illustrated in Figures 12-24 in addition to or in place of any of the multiple steps 510 of the example recipe 500. The one or more additional steps 510 can include any of the configurations of the hollow fiber system 100 described above in this disclosure (e.g., valve direction 512, pump direction 514, pump speed 516, step type 518, termination trigger threshold 520, etc.). In yet other examples, any step 510 of the recipe 500 can be repeated any number of times before proceeding to the next step 510 of the recipe 500.

[0106] It will be understood that Figures 12-24 include the same components as Figure 3, however, for clarity, only some of the components are labeled in Figures 12-24. For example, pumps that are actively pumping fluid are labeled, but pumps that are not actively pumping fluid are not labeled. Also, valves that are open are labeled, but valves that are closed are not labeled. Also, sensors that have fluid actively passing through them are labeled, but sensors that do not have fluid actively passing through them are not labeled. In these examples, the direction of the arrows indicates fluid flow within the hollow fiber system 100. Additionally, thick lines represent conduits with active fluid flow paths, while non-thick lines represent conduits that do not have active fluid flow paths. Notably, non-thick lines may still contain liquid, but are not actively flowing through them.

[0107] 12 illustrates the execution of a first step 510 of an exemplary recipe 500 for priming the intracapillary space 338 of the filter module 334 with the transduction medium 304. In the illustrated example, the controller 200 directs the hollow fiber system 100 to execute operations according to the exemplary recipe 500. Specifically, the controller 200 directs the multiple pinch valves 316 to execute the first step 510 of the exemplary recipe 500 such that the pinch valves 316a, 316g, 316h, 316k, 316n, 316q are open and all other pinch valves 316 are closed. Here, the user or the controller 200 can open the flow valves 314a, 314f. The controller 200 then directs the first pump 328 to pump the transduction medium 304 from the transduction container 302a through the intra-capillary space 338 (shown in FIG. 3B) of the filter module 334 to the waste 324 of the waste container 322a. In this example, the air bubble sensors 318a, 318g provide the controller 200 with air volume values ​​representative of the amount of air in the transduction conduit 340 and the waste conduit 350, respectively. Additionally, the first fluid sensor 320a provides the controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow at the first intra-capillary port 360a.

[0108] 13 illustrates the execution of a second step 510 of an example recipe 500 for priming the extracapillary space 339 (shown in FIG. 3B) of the filter module 334 with the transduction medium 304. In the illustrated example, the controller 200 directs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 directs the multiple pinch valves 316 to execute the second step 510 of the example recipe 500 such that the pinch valves 316a, 316g, 316l, 316m, 316o, 316q are open and all other pinch valves 316 are closed. Here, the user or the controller 200 can open the flow valves 314a, 314f. The controller 200 then directs the first pump 328 to pump the transduction medium 304 from the transduction container 302a through the extra-capillary space 339 of the filter module 334 to the waste 324 of the waste container 322a. In this example, the air bubble sensors 318a, 318g provide the controller 200 with air volume values ​​representative of the amount of air in the transduction conduit 340 and the waste conduit 350, respectively. Additionally, the second fluid sensor 320b provides the controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow at the first extra-capillary port 364a.

[0109] 14 illustrates the execution of a third step 510 of an example recipe 500 for priming the extracapillary bypass with transduction medium 304. In the illustrated example, the controller 200 directs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 directs the plurality of pinch valves 316 to execute the third step 510 of the recipe 500 such that pinch valves 316a, 316g, 316l, 316p, 316q are open and all other pinch valves 316 are closed. Now, the user or the controller 200 can open the flow valves 314a, 314f. The controller 200 then directs the first pump 328 to pump the transduction medium 304 from the transduction container 302a through the extracapillary bypass conduit to the waste 324 of the waste container 322a. In this example, air bubble sensors 318a, 318g provide air volume values ​​to controller 200 that represent the amount of air in transduction conduit 340 and waste conduit 350, respectively.

[0110] 15 illustrates the execution of the fourth step 510 of the example recipe 500 for priming the second pump 332 and the extracapillary bypass with the transduction medium 304. In the illustrated example, the controller 200 directs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 directs the plurality of pinch valves 316 to execute the fourth step 510 of the example recipe 500 such that the pinch valves 316a, 316p, 316q are open and all other pinch valves 316 are closed. Now, the user or the controller 200 can open the flow valves 314a, 314f. The controller 200 then directs the second pump 332 to pump the transduction medium 304 from the transduction container 302a through the extracapillary bypass conduit to the waste 324 of the waste container 322a. In this example, air bubble sensors 318a, 318g provide air volume values ​​to controller 200 that represent the amount of air in transduction conduit 340 and waste conduit 350, respectively.

[0111] FIG. 16 illustrates the execution of the fifth step 510 of the example recipe 500 for an integrity test. In the illustrated example, the controller 200 instructs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 instructs the plurality of pinch valves 316 to execute the fifth step 510 of the example recipe 500, as do the pinch valves 316c, 316g, 316h. Here, the user or the controller 200 may open the air filter 312 to allow air to be pumped into the air filter conduit 342 (shown in FIG. 1C). The controller 200 then instructs the first pump 328 to pump air into the air filter 312 to purge the transduction medium 304 from the filter module 334. That is, the controller 200 can open the pinch valves 316k, 316n, 316q and the flow valve 314f to pump air through the filter module 334 and drain the transduction medium 304 into the waste container 322a (this configuration is not shown in FIG. 16). The controller 200 can then close the pinch valves 316k, 316n, 316q and pressurize the intracapillary space 338 (shown in FIG. 3B) to a first pressure target (e.g., 15 PSI). Here, the first fluid sensor 320a can indicate a pressure value. After reaching the pressure target, the controller 200 can stop the operation of the first pump 328 and continue acquiring and monitoring the pressure at the first fluid sensor 320a. Additionally, the controller 200 acquires and monitors an air content value at the air bubble sensor 318 to determine whether air bubbles are present. If the pressure drop exceeds a threshold and / or if the air content value exceeds a threshold, the integrity test fails.

[0112] 17 illustrates the execution of the sixth step 510 of the example recipe 500 for priming the cell conduit 344 with a fluid after loading the cell container 302b. Thus, the welder 190 can weld the cell container 302b to the cell conduit 344 to fluidly connect the cell container 302b to the hollow fiber system 100. In the illustrated example, the controller 200 directs the hollow fiber system 100 to perform operations according to the example recipe 500. Specifically, the controller 200 directs the multiple pinch valves 316 to execute the sixth step 510 of the recipe 500 such that the pinch valves 316b, 316p, 316q are opened and all other pinch valves 316 are closed. Here, the user or the controller 200 can open the flow valves 314b, 314f. The controller 200 then instructs the second pump 332 to pump waste 324 from the waste container 322a through the extracapillary bypass conduit to the cells 306 in the cell container 302b. In particular, the second pump 332 pumps fluid in a second direction from the output container 322 to the input container 302. The sixth step 510 is performed to purge air from the cell conduit 344 before loading the cells 306 into the hollow fiber system 100. Advantageously, this prevents air bubbles from being pushed into the filter module 334 along with the cells 306 that may interfere with transduction. In this example, the air bubble sensors 318b, 318g provide the controller 200 with an air content value that represents the amount of air in the cell conduit 344 and the waste conduit 350, respectively. The air bubble sensors 318b, 318g report that there is no air in the cell conduit 344, indicating that the sixth step 510 is complete.

[0113] 18 illustrates the execution of the seventh step 510 of the example recipe 500 for loading cells 306 into the filter module 334. Loading the filter module 334 refers to forcing the input materials 304, 306, 308, 310 into the intracapillary space 338 and / or extracapillary space 339 of the filter module 334. In some embodiments, the mixer 195 mixes the cells 306 with the vector 310 and loads the mixture into the filter module 334 as described with reference to FIG. 24. In the illustrated example, the controller 200 directs the hollow fiber system 100 to perform operations according to the example recipe 500. Specifically, the controller 200 instructs the plurality of pinch valves 316 to execute the seventh step 510 of the example recipe such that pinch valves 316b, 316g, 316h, 316i, 316j, 316k, 316m, 316o, 316p, 316q are open and all other pinch valves 316 are closed. Now, the user or the controller 200 can open the flow valves 314b, 314f. The controller 200 then instructs the first pump 328 to pump the cells 306 (or a mixture of cells 306 and vectors 310 as described in FIG. 24 ) from the cell container 302b through the intracapillary space 338 and extracapillary space 339 of the filter module 334 to the waste 324 of the waste container 322a.

[0114] More specifically, the first pump 328 pumps the cells 306 (or a mixture of cells 306 and vectors 310 as described in FIG. 24 ) into both the first intracapillary port 360a and the second intracapillary port 360b, creating a counterflow of cells within the intracapillary space 338. As the cells 306 flow from both ends of the intracapillary space 338 of the filter module 334, the counterflow of cells 306 collide and / or coalesce at a common area within the intracapillary space 338, trapping the cells 306 within the filter module 334. Additionally, the fluid in which the cells 306 are suspended can flow from the intracapillary space 338 to the extracapillary space 339, thereby discharging the fluid from the extracapillary space 339 through the first extracapillary port 364a and the second extracapillary port 364b, through the waste conduit 350, and into the waste container 322a. In particular, cells 306 (or a mixture of cells 306 and vectors 310 as described in FIG. 24) do not flow from intracapillary space 338 to extracapillary space 339, but rather only the fluid in which cells 306 are suspended.

[0115] In this example, the air bubble sensors 318a, 318g provide the controller 200 with air volume values ​​representative of the amount of air in the transduction conduit 340 and the waste conduit 350, respectively. Additionally, the first fluid sensor 320a provides the controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow of the cells 306 into the first intra-capillary port 360a. The second fluid sensor 320b provides the controller 200 with pressure and / or fluid values ​​representative of the pressure and / or fluid flow of the cells 306 flowing from the first extra-capillary port 364a.

[0116] 24 illustrates the execution of the optional mixing step 510 of the example recipe 500 to transport the vector 310 from the vector container 302d to the cell container 302b, which may be performed prior to loading the filter module 334, as described with respect to FIG. 18. In the illustrated example, the controller 200 instructs the hollow fiber system 100 to perform operations according to the example recipe 500. Specifically, the controller 200 instructs the multiple pinch valves 316 to execute the mixing step 510 of the example recipe 500 such that the pinch valves 316b, 316f are open and all other pinch valves 316 are closed. Now, the user or controller can open the flow valves 314b, 314e. The controller 200 then instructs the second pump 332 to pump the population of vector particles (e.g., the vectors 310) from the vector container 302d to the cell container 302b containing the population of cells 306. In particular, when vector particles are delivered to cell container 302b, vectors 310 bind to cells 306 to produce a mixture thereof.

[0117] In some examples, the controller 200 instructs the mixer 195 to mix the cells 306 and the vectors 310. That is, the mixer 195 is in fluid communication with the cell container 302b and is configured to mix the cells 306 and the vectors 310. The controller 200 can then instruct the mixer 195 to return the mixture to the cell container 302b. Alternatively, the user may manually facilitate mixing of the cells 306 and the vectors 310 in the cell container 302b by manipulating or massaging the cell container 302b.

[0118] FIG. 19 illustrates the execution of the eighth step 510 of the example recipe 500 for transduction medium 304 tracing to remove cells 306 from the conduit. That is, the recipe of FIG. 19 is executed after the execution of the sixth step 510 of the example recipe 500 (FIG. 18) to remove remaining cells 306 from the conduit to the filter module 334. Removing the remaining cells prevents accumulation or precipitation of cells in the conduit. In the illustrated example, the controller 200 instructs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 instructs the plurality of pinch valves 316 to execute the eighth step 510 of the example recipe 500 such that pinch valves 316a, 316g, 316h, 316i, 316j, 316k, 316m, 316o, 316p, 316q are open and all other pinch valves 316 are closed. The user or the controller 200 can now open the flow valves 314a, 314f. The controller 200 then directs the first pump 328 to pump the transduction medium 304 from the transduction container 302a through the intracapillary space 338 and the extracapillary space 339 of the filter module 334 to the waste 324 of the waste container 322a. The flow of the transduction medium 304 drives the remaining cells 306 into the filter module 334.

[0119] More specifically, the first pump 328 pumps the transduction medium 304 through both the first intra-capillary port 360a and the second intra-capillary port 360b into the intra-capillary space 338 (see FIG. 3B). The transduction medium 304 flows from the intra-capillary space 338 through the porous membrane to the extra-capillary space 339 (see FIG. 3B), exits the filter module 334 through the first extra-capillary port 364a and the second extra-capillary port 364b, and is discharged through the waste conduit 350 into the waste container 322a. In this example, the air bubble sensors 318a, 318g provide air volume values ​​to the controller 200 that are representative of the amount of air in the transduction conduit 340 and the waste conduit 350, respectively. Additionally, the first fluid sensor 320a provides the controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow of the transduction medium 304 into the first intra-capillary port 360a. The second fluid sensor 320b provides the controller 200 with pressure and / or fluid values ​​representative of the pressure and / or fluid flow of the transduction medium 304 flowing from the first extra-capillary port 364a.

[0120] 20 illustrates the execution of the ninth step 510 of the example recipe 500 for loading the vector 310 into the filter module 334. In some embodiments, the mixer 195 mixes the cells 306 with the vector 310 prior to loading the cells 306 into the filter module 334. In the illustrated example, the controller 200 directs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 directs the multiple pinch valves 316 to execute the ninth step 510 of the example recipe 500 such that the pinch valves 316e, 316g, 316h, 316i, 316j, 316k, 316m, 316o, 316p, 316q are open and all other pinch valves 316 are closed. Here, the user or the controller 200 can open the flow valves 314d, 314f. The controller 200 then directs the first pump 328 to pump the vector 310 from the vector container 302d through the intra-capillary space 338 and the extra-capillary space 339 of the filter module 334 and into the waste 324 of the waste container 322a.

[0121] More specifically, the first pump 328 pumps the vector 310 into both the first intracapillary port 360a and the second intracapillary port 360b, creating a counterflow of the vector 310 (as shown in FIG. 3B) within the intracapillary space 338. As the vector 310 flows from both ends of the intracapillary space 338 (see FIG. 3B) of the filter module 334, the counterflows of the vector 310 collide and / or coalesce at a common region within the intracapillary space 338, trapping the vector 310 within the filter module 334. The common region may be the same common region of the cells 306 (e.g., seventh step 510), such that the common region defines a transduction zone. Additionally, the fluid in which the vectors 310 are suspended may flow from the intracapillary space 338 to the extracapillary space 339, thereby discharging the extracapillary space 339 through the first and second extracapillary ports 364a, 364b, through the waste conduit 350, and into the waste container 322a. In particular, the vectors 310 remain within the intracapillary space 338, and only the fluid in which the vectors 310 are suspended flows from the intracapillary space 338 to the extracapillary space 339 (e.g., as shown by the cross-flow in FIG. 3B).

[0122] In this example, the air bubble sensors 318a, 318g provide the controller 200 with air volume values ​​representative of the amount of air in the transduction conduit 340 and the waste conduit 350, respectively. Additionally, the first fluid sensor 320a provides the controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow of the vector 310 into the first intra-capillary port 360a. The second fluid sensor 320b provides the controller 200 with pressure and / or fluid values ​​representative of the pressure and / or fluid flow of the vector 310 flowing from the first extra-capillary port 364a.

[0123] In some embodiments, the controller 200 loads the cells 306 and the vectors 310 into the filter module 334 sequentially. That is, the controller 200 can first load the cells 306 into the filter module 334, and then load the vectors 310 into the filter module 334 (e.g., perform the seventh step 510, then perform the ninth step 510). Alternatively, the controller can first load the vectors 310 into the filter module 334, and then load the cells 306 into the filter module 334 (e.g., perform the ninth step 510, then perform the seventh step 510).

[0124] In some embodiments, the controller 200 may alternate between loading the cells 306 into the filter module 334 (e.g., seventh step 510) and loading the vectors 310 into the filter module (e.g., ninth step 510). That is, loading the cells 306 into the filter module 334 may create a first layer of cells 306 in the intracapillary space 338 of the filter module 334. Then loading the vectors 310 into the filter module 334 creates a first layer of vectors (e.g., on top of the first layer of cells). The controller 200 continues loading the cells 306 and the vectors 310 alternately into the filter module 334 to create multiple alternating layers of cells 306 and multiple layers of vectors 310 before harvesting the cells 306 (FIG. 23). Thus, in a series of repeated steps in an example recipe, loading cells 306 into filter module 334 may correspond to a first step in the series of repeated steps in the example recipe, and loading vectors 310 into filter module corresponds to a second step in the series of repeated steps (or vice versa). Alternating layers of cells 306 and vectors 310 can increase collisions between cells 306 and vectors 310 during transduction.

[0125] 21 illustrates the execution of the tenth step 510 of the exemplary recipe 500 during the transduction period of the cells 306 and the vectors 310. That is, the tenth step 510 can be executed after the execution of the ninth step 510 to facilitate the transduction of the cells 306 and the vectors 310 loaded in the filter module 334. In the illustrated example, the controller 200 instructs the hollow fiber system 100 to execute operations according to the recipe. Specifically, the controller 200 instructs the multiple pinch valves 316 to execute the tenth step 510 of the exemplary recipe 500 such that the pinch valves 316a, 316g, 316h, 316i, 316j, 316k, 316m, 316o, 316p, 316q are opened and all other pinch valves 316 are closed. Here, the user or the controller 200 can open the flow valves 314a, 314f. The controller 200 then directs the first pump 328 to pump the transduction medium 304 from the transduction container 302a through the intracapillary space 338 and extracapillary space 339 (shown in FIG. 3B) of the filter module 334 to the waste 324 of the waste container 322a. The flow of the transduction medium 304 prevents diffusion and forces the cells 306 and vectors 310 to co-localize on the surface of the filter module 334, thereby increasing transduction efficiency.

[0126] More specifically, the first pump 328 pumps the transduction medium 304 through both the first intra-capillary port 360a and the second intra-capillary port 360b into the intra-capillary space 338 (see FIG. 3B). The transduction medium 304 flows from the intra-capillary space 338 through the porous membrane to the extra-capillary space 339 (see FIG. 3B), exits the filter module 334 through the first extra-capillary port 364a and the second extra-capillary port 364b, and is discharged through the waste conduit 350 into the waste container 322a. In this example, the air bubble sensors 318a, 318g provide air volume values ​​to the controller 200 that are representative of the amount of air in the transduction conduit 340 and the waste conduit 350, respectively. Additionally, the first fluid sensor 320a provides the controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow of the transduction medium 304 into the first intra-capillary port 360a. The second fluid sensor 320b provides the controller 200 with pressure and / or fluid values ​​representative of the pressure and / or fluid flow of the transduction medium 304 flowing from the first extra-capillary port 364a.

[0127] 22 illustrates the execution of an eleventh step 510 of the example recipe 500 for loading the filter module 334 with the vector 310, which may be executed in addition to or instead of the ninth step 510 of the example recipe 500. In the illustrated example, the controller 200 directs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 directs the multiple pinch valves 316 to execute the eleventh step 510 of the example recipe 500 such that the pinch valves 316e, 316f, 316g, 316h, 316i, 316j, 316k, 316m, 316o, 316p are opened and all other pinch valves 316 are closed. Here, the user or the controller 200 may open the flow valves 314d, 314e.

[0128] The controller 200 then directs the first pump 328 to pump the vector 310 from the vector container 302d through the first viral conduit 348a to both the first intracapillary port 360a and the second intracapillary port 360b to create a counterflow of the vector 310 within the intracapillary space 338. As the vector 310 flows from both ends of the intracapillary space 338 of the filter module 334, the counterflow of the vector 310 collides and / or coalesces at a common area within the intracapillary space 338, trapping the vector 310 within the filter module 334. The common area may be the same common area of ​​the cells 306 (FIG. 19), such that the common area defines a transduction zone. Additionally, the fluid in which the vectors 310 are suspended may flow from the intracapillary space 338 to the extracapillary space 339, and thereby be discharged from the extracapillary space 339 through the first extracapillary port 364a and the second extracapillary port 364b, through the second viral conduit 348b, and into the vector container 302d. In this manner, the vectors 310 circulate from the vector container 302d to the filter module 334 and back again, increasing the likelihood that the vectors 310 will interact with the cells 306 disposed within the filter module 334.

[0129] In this example, air bubble sensors 318e, 318f provide controller 200 with air volume values ​​representative of the amount of air in first and second viral conduits 348a, 348b, respectively. Additionally, first fluid sensor 320a provides controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow of vector 310 into first intra-capillary port 360a. Second fluid sensor 320b provides controller 200 with pressure and / or fluid values ​​representative of the pressure and / or fluid flow of vector 310 flowing from first extra-capillary port 364a.

[0130] 23 illustrates the execution of the twelfth step 510 of the example recipe 500 during a harvest period, where the harvest container 322b collects the cells 306 from the filter module 334 after the transduction period. In the illustrated example, the controller 200 instructs the hollow fiber system 100 to execute operations according to the example recipe 500. Specifically, the controller 200 instructs the multiple pinch valves 316 to execute the twelfth step 510 of the example recipe 500 such that the pinch valves 316d, 316g, 316j, 316k, 316m, 316o, 316p, 316r are open and all other pinch valves 316 are closed. Here, the user or the controller 200 can open the flow valves 314c, 314g. The controller 200 then causes the first pump 328 and the second pump 332 to pump the culture medium 308 from the culture container 302c, through the intra-capillary space 338 and the extra-capillary space 339 of the filter module 334, and into the collection medium 326 of the collection container 322b.

[0131] More specifically, the first pump 328 pumps the culture medium 308 through the intracapillary space 338 to the first intracapillary port 360a, and through the collection conduit 352 to the second intracapillary port 360b to the collection container 322b. Furthermore, the second pump 332 pumps the culture medium 308 through the first extracapillary port 364a and the second extracapillary port 364b. Thus, the culture medium 308 flows from the extracapillary space 339 to the intracapillary space 338 through the porous membrane. Thus, the culture medium 308 exits the filter module 334 through the second extracapillary port 364b to the collection conduit 352, and is finally pumped to the collection container 322b. Notably, here, the fluid flow direction of the culture medium 308 is from the extracapillary port 364 to the intracapillary space 338 of the filter module 334. This reverse fluid flow lifts the cells 306 from the membrane into the intracapillary space 338. Additionally, flow from the first intracapillary port 360a to the second intracapillary port 360b drives the cells 306 into the collection container 322b.

[0132] In this example, air bubble sensors 318d, 318h provide controller 200 with air volume values ​​representative of the amount of air in culture conduit 346 and collection conduit 352, respectively. Additionally, first fluid sensor 320a provides controller 200 with pressure and / or fluid flow values ​​representative of the pressure and / or fluid flow of culture medium 308 to first intra-capillary port 360a. Second fluid sensor 320b provides controller 200 with pressure and / or fluid values ​​representative of the pressure and / or fluid flow of culture medium 308 flowing to first extra-capillary port 364a.

[0133] 25, a schematic diagram of another example of hollow fiber system 100a is provided. Because the structure and function of components associated with hollow fiber system 100 are substantially similar to hollow fiber system 100a, like reference numbers are used below and in the drawings to identify like components, and like reference numbers with letter extensions are used to identify modified components or components that are provided in multiple instances.

[0134] 25 includes common components with the hollow fiber system 100 described above, however, the hollow fiber system 100a has been modified to include alternative flow paths and valves configured to pass input materials 304, 306, 308, 310 through the hollow fiber system 100a. For example, the transduction conduit 340a of the hollow fiber system 100a branches into a first transduction sub-conduit 341a and a second transduction sub-conduit 341b, where the first transduction sub-conduit 341a provides a direct fluid path from the transduction container 302a to the first pump inlet 327, while the second transduction sub-conduit 341b is connected to conduits 342, 344, 346, 348 upstream of the first pump inlet 327 and the second pump inlet 331, respectively. Thus, the first transduction subconduit 341a and the second transduction subconduit 341b provide parallel and alternative flow paths between the transduction container 302a and at least the first pump 328. Each of the first transduction subconduit 341a and the second transduction subconduit 341b may include one or more valves 317. In FIG. 25, each valve 317 is depicted generically. However, the one or more valves 317 may include flow valves 314, pinch valves 316, and / or other types of valves for controlling flow through the transduction subconduits 341a, 341b. Optionally, each transduction subconduit 341a, 341b may include a flow sensor, such as an air bubble sensor 318.

[0135] 25, hollow fiber system 100a is provided with an alternative configuration of second intra-capillary conduit 356c. While second intra-capillary conduit 356b described above connects first pump outlet 329 to second intra-capillary port 360b, second intra-capillary conduit 356c of hollow fiber system 100a provides a direct fluid flow path from second pump outlet 333 to second intra-capillary port 360b. Second intra-capillary conduit 356c may include one or more valves 317, which may include flow valve 314 and / or pinch valve 316. Optionally, second intra-capillary conduit 356c may include a flow sensor, such as air bubble sensor 318.

[0136] In an alternative configuration of the second intra-capillary conduit 356c, the extra-capillary conduit 368 may include one or more valves 317 for selectively controlling flow to both the first extra-capillary sub-conduit 368a and the second extra-capillary sub-conduit 368b. However, in some examples, the valves 317 may be omitted from the extra-capillary conduit 368, such that flow through each of the extra-capillary sub-conduits 368a, 368b is independently controlled via corresponding pinch valves 316m, 316o, 316p.

[0137] 26-46 illustrate example schematics of fluid flow paths through hollow fiber system 100a according to a plurality of steps 510 of recipe 500. Specifically, each of FIGS. 26-46 illustrates one of the plurality of steps 510 of recipe 500. It should be understood that the plurality of steps 510 depicted in FIGS. 26-46 are merely illustrative examples of recipes and are not intended to limit the present disclosure in any way. For example, different example recipes may repeat any of the plurality of steps any number of times, include more or fewer steps, and / or perform the plurality of steps in a different order than the example recipes illustrated in FIGS. 26-46.

[0138] It will be understood that Figures 26-46 include the same components as Figure 25, however, for clarity, all components of system 100a are labeled in Figure 26. In these examples, the direction of the arrows indicates fluid flow within hollow fiber system 100a. Additionally, bold lines represent conduits with active fluid flow paths, while non-bold lines represent conduits that do not have active fluid flow paths. Notably, non-bold lines may still contain liquid, but are not actively flowing through them.

[0139] The configuration of hollow fiber system 100a provided in Figures 25-46 provides different functionality than hollow fiber system 100 shown in Figures 3 and 12-24. For example, by providing a first transduction sub-conduit 341a and a second intracapillary conduit 356c, hollow fiber system 100a is operable to prime all components of the system with transduction medium 304, as shown in Figures 26-31, 35, and 36. This ensures that clean transduction medium 304 always flows through the conduits, rather than relying on backflow of waste 324 from waste container 322a. For example, compare the performance of the sixth step 510 for priming cell conduit 344 shown in Figure 17 with the priming of cell conduit 344 shown in Figure 36. In FIG. 17, the cell conduit 344 is primed using waste 324 from the waste container 322, whereas in FIG. 36, the cell conduit 344 is primed using transduction medium 304 (i.e., priming medium) via the first transduction sub-conduit 341a.

[0140] Providing a forward flow of transduction medium 304 to prime the cell flow path, rather than utilizing a reverse flow of waste 324, is particularly useful for ensuring compliance with Good Manufacturing Practices (GMP) since the transduction medium 304 is free of contaminants that may be flushed from the system 100a into the waste container 322a during any of the priming steps. For example, the filter module 334 may initially be provided by the manufacturer with a sugar substrate that is included during shipment and storage of the filter module 334 to stabilize the pores. In the priming step, the sugar substrate is flushed from the filter module 334, preparing the filter module 334 for use. By utilizing the transduction medium only as a priming solution, the system 100a eliminates the possibility of introducing sugar substrates that may be included in the waste 324 into the cell container 302b during the priming step of the cell conduit 344.

[0141] In addition to providing a different priming configuration, hollow fiber system 100a provides a more direct flow path between input container 302 and second intracapillary port 360b by providing second intracapillary conduit 356c. In particular, as shown in FIG. 38, providing second intracapillary conduit 356c allows cells 306 to be delivered directly to second intracapillary port 360b via second pump 332. Thus, rather than relying on first pump 328 to supply cells 306 to both intracapillary ports 360a, 360b, cells 306 can be independently supplied to intracapillary ports 360a, 360b by each one of pumps 328, 332 (see, e.g., FIG. 18). This configuration improves system efficiency and provides redundancy of components within hollow fiber system 100a.

[0142] 25-45, the implementation of hollow fiber system 100a is shown generally with respect to the arrangement of components provided for hollow fiber system 100 shown in FIG. 1A, however, the configuration of hollow fiber system 100a advantageously allows the length of some conduits to be minimized relative to the length of such conduits in system 100. For example, by implementing second intracapillary conduit 356c flowing directly from second pump 332, the overall length of second intracapillary conduit 356c is reduced relative to the length of second intracapillary conduit 356b in hollow fiber system 100. Similarly, reconfiguring intracapillary conduit 356c allows pinch valve 316j (see FIG. 18) to be removed from system 100a. Eliminating this pinch valve 316j allows collection conduit 352a to be reduced relative to the length of collection conduit 352 provided in system 100. By designing the system 100a to minimize the length of each conduit, areas of low flow or stagnation within the system 100a are eliminated, thereby minimizing the chance of cells becoming clogged within the conduits and / or settling outside the filter module 334 during the cell loading step of the recipe 500.

[0143] FIG. 46 illustrates a chart 4600 of an optimization study in a fully automated hollow fiber system 100. The chart 4600 demonstrates retroviral transduction of cells 306 in the hollow fiber system 100. In this example, the cells 306 are αβ T cells. The chart 4600 illustrates a comparison of the results of a first condition 4602, a second condition 4604, and a third condition 4606 for loading the cells 306 and the vector 310 into the filter module 334. The first condition 4602 includes untransduced cells. The second condition 4604 includes the cells 306 and the vector 310 loaded into the filter module 334 under a constant pressure, and the third condition 4606 includes the cells 306 and the vector 310 loaded into the filter module 334 under a constant flow rate. A second condition 4604 is configured such that the hollow fiber system 100 adjusts the flow rate of the pumps 328, 332 to maintain a target fluid pressure. A third condition 4606 is configured such that the fully automated hollow fiber system 100 is configured to maintain the flow rate of the pumps 328, 332 at a pre-set speed. For example, the third condition 4606 may cause the hollow fiber system 100 to adjust the flow rate of the pumps 328, 332 only if the hollow fiber system 100 reaches or exceeds a threshold pressure setting.

[0144] FIG. 47 illustrates a chart 4700 of an optimization study in a fully automated hollow fiber system 100 platform. The chart 4700 demonstrates retroviral transduction of αβ T cells 306 in the hollow fiber system 100. The chart 4700 illustrates a comparison of results associated with a first condition 4702, a second condition 4704, and a third condition 4706 for loading of cells 306 with vector 310. The first condition 4702 includes untransduced cells 306. The second condition 4704 includes approximately 1 mole (M) cells / cm in a hollow fiber approximately 20 centimeters long. 2 The third condition 4706 includes cells 306 and vectors loaded at a concentration of about 1M cells / cm 2The cells 306 and vector 310 were loaded into a 40 centimeter length hollow fiber at a concentration of 0.1 μg / mL. During operation, the flux rate, cell number, and cell volume were normalized across the length of the hollow fiber for the second and third conditions 4704, 4706 based on the relative difference in surface area across the two fibers.

[0145] FIG. 48 illustrates a chart 4800 of an optimization study in a fully automated hollow fiber system 100. The chart 4800 demonstrates retroviral transduction of αβ T cells 306 in the hollow fiber system 100. The chart 4800 illustrates a comparison of results associated with a first condition 4802, a second condition 4804, and a third condition 4806 for cells 306 and vector 310 loaded into a filter module 334. The first condition 4802 includes untransduced cells 306. The second condition 4804 includes premixing the cells 306 and vector 310 on the hollow fiber system 100 before loading the mixture into the filter module 334. The third condition 4800 includes sequential loading of the cells 306 and vector 310 into the filter module 334. In a third condition 4806, the cells 306 are loaded into the hollow fiber system 100 first, and then the vector is loaded.

[0146] In a second condition 4804, the fully automated hollow fiber system 100 pumps the vectors 310 from the vector transfer bag 302 into the transfer bag 302 containing the cell suspension 306. The cells 306 are then manually mixed by an operator before the mixed solution of cells 306 and vectors 310 is loaded into the filter module 334. In a third condition 4806, the cells 306 and vectors 310 remain separated during the loading step and only coexist within the filter module 334. The flow rate is believed to remain constant throughout the second and third conditions 4804, 4806.

[0147] Charts 4600, 4700, and 4800 each illustrate an example of a gamma retroviral vector 310 using a chimeric antigen receptor (CAR) construct. CD4 / CD8 isolated T cells were thawed and activated for 48 hours. CD4 T cells are a type of white blood cell that helps fight infections, while CD8 T cells help eliminate virus-infected cells. Operations to obtain the results shown in charts 4600, 4700, and 4800 include the following operations: After thawing and activating the cells 306, the cells 306 and the vector 310 are transferred to separate transfer bags 302, which are sterilized and welded to the hollow fiber system 100. Further operations may involve starting a predefined program on the computing device 200 of the hollow fiber system 100 to introduce the cells 306 and the vector 310 under constant pressure or constant flow conditions. Both conditions are transduced at a multiplicity of infection (MOI) of about 5. It is contemplated that alternative operations may involve an MOI greater than or less than 5. Once the cells 306 and vector 310 are introduced, the hollow fiber system 100 automatically transitions to a 90 minute transduction phase followed by a harvest operation to harvest the cells 306 from the filter module 334. The harvested cells 306 are cultured for an additional period of time (approximately 3-4 days) before analyzing the percentage of CAR expression using flow cytometry.

[0148] 49 illustrates a chart 4900 of an optimization study in a fully automated hollow fiber system 100. The chart 4900 demonstrates the results of retroviral transduction of NK cells 306 in a filter module 334. The chart 4900 illustrates a comparison of results associated with a first condition 4902 and a second condition 4904 for cells 306 and vector 310 loaded into the filter module 334. The first condition 4902 includes untransduced cells 306 and the second condition 4904 includes NK cells 306 transduced in the fully automated hollow fiber system 100.

[0149] In this example, the vector 310 may be a gamma retroviral vector using a CAR construct. During operation, the CD56 NK cells 306 are negatively isolated and activated for about 6 days. CD56 is a phenotypic marker for NK cells. The cells 306 and vector 310 are then transferred to separate transfer bags 302, which are sterile welded to the hollow fiber system 100. A predefined program may then be initiated on the computing device 200 of the hollow fiber system 100 to introduce the cells 306 and vector 310 under conditions of constant pressure or constant flow. As previously mentioned, both conditions are transduced at an MOI of about 5. It is contemplated that in alternative operations, the MOI may be greater or less than 5. After introduction of the cells 306 and vector 310, the platform may automatically proceed to a 90 minute transduction phase, followed by a harvest step to recover the cells 306 from the filter module 334. The recovered cells 306 may be reactivated and cultured for an additional period of time (approximately 14 days) before analyzing the percentage of CAR expression using flow cytometry.

[0150] FIG. 50 illustrates a chart 5000 of an optimization study in a fully automated hollow fiber system 100. The chart 5000 demonstrates the results of retroviral transduction of γδ T cells 306 in a filter module 334. The chart 5000 illustrates a comparison of results associated with a first condition 5002 and a second condition 5004 for cells 306 and vectors 310 loaded into the filter module 334. The first condition 5002 includes untransduced cells 306 and the second condition 5004 includes γδ T cells 306 transduced with the fully automated hollow fiber platform 100.

[0151] In this example, the vector 310 may be a gamma retroviral vector using a CAR construct. In operation, the γδ T cells 306 are negatively isolated and activated for about 6 days. The cells 306 and the vector 310 are then transferred to separate transfer bags 302, which are sterile welded to the hollow fiber system 100. A predefined program may then be initiated on the computing device 200 of the hollow fiber system 100 to introduce the cells 306 and the vector 310 under conditions of constant pressure or constant flow. Both conditions are transduced at an MOI of about 7.5. It is contemplated that in alternative operations, the MOI may be greater or less than 7.5. After introduction of the cells 306 and the vector 310, the platform may automatically move to a 90 minute transduction phase, followed by a harvest step to collect the cells 306 from the filter module 334. The collected cells 306 may be reactivated and cultured for an additional period (about 10 days) before analyzing the percentage of CAR expression using flow cytometry.

[0152] Although several embodiments have been described, it will be understood that various modifications may be made without departing from the spirit and scope of the disclosure. Accordingly, other embodiments are within the scope of the following claims.

Claims

1. 1. A computer-implemented method that, when executed on data processing hardware, causes the data processing hardware to perform operations for executing a recipe with a hollow fiber transduction device having a filter module, a pump, and a plurality of sensors, the recipe including a plurality of steps, each step including a first termination trigger threshold indicating whether the respective step of the recipe is complete, the operations comprising: directing the pump to execute one of the steps of the recipe to provide a first fluid flow having a first pressure at a first sensor of the plurality of sensors, the first pressure being specified in the recipe and the first sensor being positioned upstream of the filter module relative to a direction of the first fluid flow; receiving a fluid flow signal from a first sensor of the plurality of sensors indicative of an operating parameter measured at the first sensor; comparing a first termination trigger threshold for the one of the steps of the recipe to the received fluid flow signal, the comparison of the first termination trigger threshold indicating whether an operating condition has been met; instructing the pump to complete the one of the steps of the recipe based on a comparison of the first termination trigger threshold; The computer-implemented method comprising:

2. Each step of the recipe is: Valve configuration, Pump direction, Pump speed, or Step type, The computer-implemented method of claim 1 , comprising at least one of:

3. 3. The computer-implemented method of claim 1, wherein instructing the pump to perform the one of the steps of the recipe providing the first fluid flow having a first pressure at the first sensor of the plurality of sensors includes sending a pump speed signal to the pump to operate at a first pump speed.

4. 4. The computer-implemented method of claim 3, wherein instructing the pump to perform the one of the steps of the recipe providing the first fluid flow having a first pressure at the first sensor of the plurality of sensors further comprises sending a pump direction signal to the pump to operate in a first direction.

5. after instructing the pump to complete the one of the steps of the recipe based on a comparison of the first termination trigger threshold; directing the pump to execute another one of the steps of the recipe to provide a second fluid flow having a second pressure at the first sensor of the plurality of sensors; 5. The computer-implemented method of claim 4, wherein the other one of the steps of the recipe further instructs the pump to operate in a second direction opposite to the first direction.

6. The operation includes: if the received fluid flow signal does not satisfy the first termination trigger threshold for the one of the steps of the recipe, waiting a time corresponding to a timer value; receiving a second fluid flow signal from a first sensor of the plurality of sensors, the second fluid flow signal being indicative of the operating parameter measured at the first sensor; comparing the first termination trigger threshold for the one of the steps of the recipe to the received second fluid flow signal; instructing the pump to re-execute the one of the steps of the recipe in response to the comparison; The computer-implemented method of any one of claims 1 to 5, further comprising:

7. The operating parameters are: Pressure value, or A fluid flow value.

8. The computer-implemented method of any of claims 1 to 7, wherein the operations further comprise determining whether the one of the steps of the recipe is a final step of the recipe.

9. 10. The computer-implemented method of claim 8, wherein the actions further comprise: completing execution of the recipe if the one of the steps of the recipe is the last step of the recipe.

10. 10. The computer-implemented method of claim 8, wherein the actions further comprise: if the one of the steps of the recipe is not the last step of the recipe, executing a next step of the recipe.

11. 11. The computer-implemented method of claim 1, wherein the one of the steps of the recipe includes a second termination trigger threshold that indicates whether a fault occurs in the hollow fiber transduction device during execution of the respective step of the recipe.

12. Each of the first termination trigger threshold and the second termination trigger threshold is Pressure value, Timer value, Air volume value, Fluid flow value, or and a liquid to air ratio.

13. The operation includes: comparing a second termination trigger threshold for the one of the steps of the recipe to the received fluid flow signal; instructing the pump to ignore the one of the steps of the recipe based on a comparison of the second termination trigger threshold; 13. The computer-implemented method of claim 11 or claim 12, further comprising:

14. The computer-implemented method of any of claims 1 to 13, wherein the operations further comprise storing the recipe in memory hardware in communication with the data processing hardware.

15. The operation includes: receiving a request to execute the recipe stored in the memory hardware; Reading a recipe stored in the memory hardware; executing the recipe by the data processing hardware; The computer-implemented method of claim 14 further comprising:

16. The operation includes: receiving a request to modify at least one step of the recipe stored in the memory hardware; updating the at least one step of the recipe stored in the memory hardware; 16. The computer-implemented method of claim 14 or claim 15, further comprising:

17. a filter module defining an intra-capillary space and an extra-capillary space separated from the intra-capillary space by a porous membrane, the filter module including a pair of intra-capillary ports fluidly coupled to the intra-capillary space and a pair of extra-capillary ports coupled to the extra-capillary space; a first pump including a first pump inlet in communication with each of a cell source having cells and a vector source having vectors, and a first pump outlet in selective communication with each of the pair of intra-capillary ports and the pair of extra-capillary ports; a second pump including a second pump inlet in communication with each of the cell source and the vector source, and a second pump outlet in selective communication with each of the pair of intra-capillary ports and the pair of extra-capillary ports; a graphical user interface (GUI) in operative communication with each of the first pump and the second pump and configured to receive requests from a user to operate the first pump and the second pump according to a recipe including one or more steps; A system comprising:

18. at least one valve disposed between the first pump outlet and a first intra-capillary port of the pair of intra-capillary ports; at least one valve disposed between the first pump outlet and a second intra-capillary port of the pair of intra-capillary ports; a fluid sensor disposed between the first pump outlet and the first of the pair of intra-capillary ports; The system of claim 17 , wherein the valve operates according to the recipe.

19. a valve disposed between the first pump inlet and the source of cells; an air bubble sensor disposed between the first pump inlet and the source of cells; 19. The system of claim 17 or claim 18, wherein the valve operates according to the recipe.

20. at least one valve disposed between the second pump outlet and a first intra-capillary port of the pair of extra-capillary ports; at least one valve disposed between the second pump outlet and a second intra-capillary port of the pair of intra-capillary ports; a fluid sensor disposed between the second pump outlet and the first intra-capillary port of the pair of extra-capillary ports; The system of any one of claims 17 to 19, wherein the valve operates according to the recipe.

21. The system of any of claims 17 to 20, wherein the GUI is further configured to, in response to receiving the request, display the recipe from memory hardware to the user.

22. a filter module defining an intra-capillary space and an extra-capillary space separated from the intra-capillary space by a porous membrane, the filter module including a pair of intra-capillary ports fluidly coupled to opposite ends of the intra-capillary space and a pair of extra-capillary ports coupled to opposite ends of the extra-capillary space; a first pump including a first pump inlet in communication with each of a cell source having cells and a vector source having vectors, and a first pump outlet in selective communication with a first intra-capillary port of the pair of intra-capillary ports and a second intra-capillary port of the pair of intra-capillary ports; a first fluid sensor disposed between the first pump outlet and the first intra-capillary port of the pair of intra-capillary ports; Data processing hardware; memory hardware in communication with the data processing hardware, the memory hardware storing instructions that, when executed on the data processing hardware, cause the data processing hardware to operate according to a recipe for transducing the cell with the vector; A system comprising:

23. The operation includes: operating the first pump at a first pumping rate to provide a first flow of at least one of the cells or the vectors to each of the pair of intra-capillary ports of the filter module; obtaining a first fluid flow signal from the first fluid sensor, the first fluid flow signal being indicative of the first flow of at least one of the cells or the vectors; comparing the first fluid flow signal to a first fluid flow parameter threshold; operating the first pump at a second pump rate in response to the comparison; and 23. The system of claim 22, comprising:

24. 24. The system of claim 23, wherein the actions further include initiating a pump rate adjustment procedure in response to the comparison, the pump rate adjustment procedure including ceasing operation of the first pump for a predetermined period of time.

25. 25. The system of claim 24, wherein the predetermined period of time is a fixed period of time.

26. 26. The system of claim 24 or claim 25, wherein the predetermined period is a pressure adjustment period.

27. The system of any of claims 24 to 26, wherein the pump rate adjustment step further comprises operating the first pump in a reverse flow condition.

28. 28. The system of any of claims 23-27, wherein the operations further include operating the first pump at a third pumping rate to provide a second flow of the one of the cells or the other of the vectors to each of the pair of intra-capillary ports of the filter module.

29. The operation includes: operating at the first pump rate to provide the first flow of the at least one of the cells or the vectors to each of the pair of intracapillary ports of the filter module; and operating at the third pump rate to provide the second flow of the other of the one of the cells or the vectors to each of the pair of intracapillary ports of the filter module.

30. The system of claim 28, further comprising prompting for alternation.

30. 30. The system of claim 28 or claim 29, wherein the operations further include operating the first pump at the first pump rate to provide the first flow of the at least one of the cells or the vector to each of the pair of intracapillary ports of the filter module, and then operating the first pump at the third pump rate to provide the second flow of the one of the cells or the other of the vector to each of the pair of intracapillary ports of the filter module.

31. The operation includes: obtaining a second fluid flow signal from the first fluid sensor indicative of the second flow of the one of the cells or the vectors; comparing the second fluid flow signal to a second fluid flow parameter threshold; initiating a pump rate adjustment procedure in response to said comparison; The system of any of claims 28 to 30, further comprising:

32. 32. The system of any one of claims 22 to 31, further comprising a transduction medium source in communication with the first pump inlet, the operation further comprising operating the first pump at a third pump rate to provide a third flow of the transduction medium to each of the pair of intra-capillary ports.

33. 33. The system of any of claims 22-32, further comprising a second pump including a second pump inlet in communication with each of the cell source and the vector source, and a second pump outlet in selective communication with each of the pair of extracapillary ports.

34. 34. The system of claim 33, further comprising a culture medium source containing a culture medium in communication with the first pump inlet and the second pump inlet.

35. The operation includes: operating the first pump at a fourth pumping rate to provide a fourth flow of the culture medium to one of the intracapillary ports; operating the second pump at a fifth pumping rate to provide a fifth flow of the culture medium to each of the pair of extracapillary ports; 35. The system of claim 34, further comprising:

36. The system according to any one of claims 22 to 35, wherein the vector comprises at least one of a non-viral vector or a viral vector.

37. 37. The system of any of claims 22-36, further comprising a mixer configured to receive at least one of the population of cells or the population of vector particles.

38. 38. The system of claim 37, wherein the operations further comprise mixing the population of cells and the population of vector particles using the mixer to generate a mixture and then loading the mixture into the filter module.

39. The operation includes: loading said cells and said vector into said intracapillary space to effect transduction of one or more of said cells in said intracapillary space; harvesting a cell population comprising the one or more transduced cells from the intracapillary space; The system of any of claims 22-38, further comprising producing a cell therapy product comprising the one or more transduced cells by:

40. A computer-implemented method of manufacturing a cell therapy product, the method comprising: when executed on data processing hardware, causing the data processing hardware to perform operations for executing a recipe including a plurality of steps with a hollow fiber transduction device including a pump including a pump inlet in communication with each of a cell source including cells and a vector source including vectors, and a first pump outlet in selective communication with a first intra-capillary port and a second intra-capillary port of a filter module, the operations comprising: directing the pump to execute one of the steps of the recipe to provide the cells and the vector to the first intra-capillary port and the second intra-capillary port of the filter module to transduce the cells; directing the pump to execute a subsequent step of the recipe to provide the transduced cells from the filter module to a collection container in communication with the second intra-capillary port of the filter module; The method comprising:

41. 41. The computer-implemented method of claim 40, wherein the actions further comprise growing the transduced cells in an appropriate culture medium according to steps of the recipe.

42. 42. The computer-implemented method of claim 40 or claim 41, wherein the operations further comprise cryopreserving the transduced cells after expanding the transduced cells according to the steps of the recipe.

43. 43. The computer-implemented method of any of claims 40-42, wherein at least 50%, 60%, 70%, 80%, or 90% or more of the cells provided to the filter module are transduced.

44. 44. The computer-implemented method of any of claims 40 to 43, wherein the vector comprises a viral vector or a non-viral vector.

45. 45. The computer-implemented method of any of claims 40 to 44, wherein the vector comprises a chimeric antigen receptor (CAR) construct expressed on the surface of the transduced cell.

46. 46. ​​The computer-implemented method of claim 40, wherein the actions further comprise separating the transduced cells from the untransduced cells according to steps of the recipe.

47. a filter module defining an intra-capillary space and an extra-capillary space separated from the intra-capillary space by a porous membrane, the filter module including a pair of intra-capillary ports fluidly coupled to opposite ends of the intra-capillary space and a pair of extra-capillary ports coupled to opposite ends of the extra-capillary space; a first pump including a first pump inlet in communication with each of a cell source having cells and a vector source having vectors, and a first pump outlet in selective communication with a first intra-capillary port of the pair of intra-capillary ports; a second pump including a second pump inlet in communication with each of the cell source and the vector source, and a second pump outlet in selective communication with a second intra-capillary port of the pair of intra-capillary ports; a first fluid sensor disposed between the first pump outlet and the first intra-capillary port of the pair of intra-capillary ports; Data processing hardware; memory hardware in communication with the data processing hardware, the memory hardware storing instructions that, when executed on the data processing hardware, cause the data processing hardware to operate according to a recipe for transducing the cell with the vector; A system comprising:

48. 48. The system of claim 47, wherein the first pump is operable to supply a first flow comprising at least one of the cells or the vector to the first intracapillary port, and the second pump is operable to supply a second flow comprising at least one of the cells or the vector to the second intracapillary port simultaneously as the first flow is supplied to the first intracapillary port.

49. a filter module defining an intra-capillary space and an extra-capillary space separated from the intra-capillary space by a porous membrane, the filter module including a pair of intra-capillary ports fluidly coupled to opposite ends of the intra-capillary space and a pair of extra-capillary ports coupled to opposite ends of the extra-capillary space; a source of transduction medium comprising the transduction medium; A cell source comprising cells; a vector source comprising a vector; A first pump, a first pump inlet in communication with (i) the transduction medium source via a first transduction subconduit and a second transduction subconduit parallel to the first transduction subconduit, (ii) the cell source via a cell conduit, and (iii) the vector source via a vector conduit; a first pump including a first pump outlet in selective communication with a first of the pair of intra-capillary ports; a first fluid sensor disposed between the first pump outlet and the first intra-capillary port of the pair of intra-capillary ports; Data processing hardware; memory hardware in communication with the data processing hardware, the memory hardware storing instructions that, when executed on the data processing hardware, cause the data processing hardware to operate according to a recipe for transducing the cell with the vector; A system comprising:

50. 50. The system of claim 49, wherein the system is operable (i) in a first configuration to provide a first flow of the transduction medium to the first pump inlet via the first transduction subconduit, and (ii) in a second configuration to provide a second flow of the transduction medium to the first pump inlet via the second transduction conduit.

51. 51. The system of claim 49 or 50, wherein operating the system in the first configuration enables the system to prime a first flow path of the system with the transduction medium, and operating the system in the second configuration enables the system to prime a second flow path of the system with the transduction medium.

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