System and method for affinity cell selection

Abstract

A biological fluid processing system and method of operating the system for cell affinity selection. The system having a fluid processor that operates on a fluid circuit having a processing container, a source container filled with a biological fluid, a separation chamber configured to separate the biological fluid from the source container into at least two volumes of material, and a selection column connected to the fluid processor. Modified cells for use in treating a disease, ailment, or medical condition of a patient are also disclosed. The modified cells are produced using a biological fluid processing system. A separation chamber configured to separate the biological fluid from the source container into at least two volumes of material and a processing container for collecting processed cells from the separation chamber are included. A selection column is connected to the fluid circuit for further cell affinity selection processing of the cells.

Description

SYSTEM AND METHOD FOR AFFINITY CELL SELECTIONCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 613,500 filed December 21 , 2023 and U.S. Provisional Application No. 63 / 685,540 filed August 21 , 2024, the contents of which are hereby incorporated herein by reference in their entireties.TECHNICAL FIELD

[0002] The present disclosure is generally directed to devices and methods used for affinity cell selection. More particularly, the present disclosure is directed to fluid handling systems for processing biological fluids and accomplishing cell selection.Even more particularly, the present disclosure is directed to systems and methods for modifying cells and modified cells that are produced by such fluid handling systems and for using such cells to treat various diseases, ailments, or medical conditions.BACKGROUND

[0003] The processing of biological fluid such as blood or blood components typically involves using a reusable processing apparatus (“hardware”) and a disposable fluid circuit adapted for mounting to or other association with the reusable apparatus. The fluid circuit typically includes containers such as plastic bags and associated tubing that defines a flow path through the circuit. The disposable fluid circuit may also include one or more separation devices where the biological fluid / cells can be separated into two or more components, washed, or otherwise processed.

[0004] Separation devices may separate the biological fluid based on centrifugal separation, membrane separation or other procedures, such as use of magnetic fields, for example, to select and / or separate target cells. Certain systems may use a combination of these separation devices, such as described in US Pub. Nos.2017 / 0315121 and 2018 / 0172685, and US Patent 1 1 ,478,755, which are incorporated herein by reference in their entirety.SUMMARY

[0005] The present disclosure provides a system and method for affinity cell selection within biological fluids.

[0006] A system and method are disclosed for use in affinity cell selection. The system includes a fluid handling system that utilizes a disposable fluid circuit having a cell selection column and which may be configured for use of the cell selection column manually or may be automated. The system is capable of preparing apheresis- collected (or other heterogeneous cell populations) material for cell selection by automated washing, incubation, and volume / concentration adjusting of the cellular material. The system also facilitates the removal of target / non-target cells fractions by adding selection reagents to the cell selection column and subsequent washing / resuspension of the target / non-target fraction at desired cell concentrations for downstream use.

[0007] The method is highly configurable to enable the operator to define custom values and processing parameters to accommodate a large number of applications including with respect to: different cell starting materials, different cell target / non-target cell populations, purity / recovery profiles, and selection reagents.

[0008] In a first aspect, a biological fluid processing system is provided for cell affinity selection, including a fluid processor having reusable hardware that operates on a disposable fluid circuit, the fluid circuit being connectable to a source container and having a biological fluid, a separation chamber configured to separate the biological fluid from the source container into at least two volumes of material, a processing container for collecting washed and harvested cells from the separation chamber, and a selection column connected to the fluid circuit for further cell affinity selection processing of the cells from the process container.

[0009] In a second aspect, a method of operating a biological fluid processing system for cell affinity selection is provided, the method including: obtaining a biological fluid processing system comprising a fluid processor that operates on a disposable fluid circuit that is connectable to a source container having a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a processing container connected to the fluid processor along a fluid pathway,and a selection column connected to the fluid circuit. The method further comprises conducting a pre-processing state further comprising: selecting a procedure protocol, conducting procedure setup, installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor a plurality of containers; running a fluid circuit solution prime including the separation chamber; attaching to the fluid processor the source container; and running a fluid circuit source prime. The method also comprises conducting a processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid; rinsing the source container; diluting the harvested cells; collection of the harvested cells in the processing container; draining solution from the selection column; adding antibodies to the selection column; transferring the harvested cells from the processing container into the selection column; transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste; diluting the material in the selection column; repriming the separation chamber for processing the material in the selection column; target fraction loading from the selection column into the separation chamber; washing and harvesting cells from the selection column; harvesting selected cells as a final product in at least one final container; and conducting a post-processing state further comprising; sealing the at least one final container and removing the final product; and removing the disposable fluid circuit.

[0010] In a third aspect, a method of operating a biological fluid processing system for cell affinity selection is provided, wherein the system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container filled with a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a process container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit. The method comprises pre-processing, processing, and post-processing states. The pre-processing state further comprises selecting a procedure protocol; procedure setup; installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor at least a first solution container comprising a separation / processing buffer, a second solution container comprising a resuspension buffer and a third solutioncontainer comprising a releasing buffer; running a fluid circuit solution prime; attaching to the fluid processor the source container; and running a fluid circuit source prime. The processing state further comprises processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid, rinsing of the source container, dilution of the harvested cells, collecting the harvested cells in the processing container, draining buffer solution from the selection column, diluting the material in the selection column via adding antibodies to the selection column, incubating with the antibodies in the selection column, transferring the harvested cells from the processing container into the selection column, transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste, diluting the material in the selection column via transferring biotin into the selection column, incubating, repriming the separation chamber for processing material in the selection column, loading the target fraction from the selection column into the separation chamber, washing and harvesting the cells processed in the separation chamber, rinsing the source container, harvesting selected cells as a final product in at least one final container, and diluting of the harvested cells in the final container. The postprocessing state further comprises sealing the at least one final container and removing the final product, and removing the disposable fluid circuit.

[0011] In a fourth aspect, modified cells for use in the treatment of a disease, ailment, or medical condition of a patient are provided wherein the modified cells are produced using a biological fluid processing system, including a fluid processor having reusable hardware that operates on a disposable fluid circuit, the fluid circuit being connectable to a source container and having a biological fluid, a separation chamber configured to separate the biological fluid from the source container into at least two volumes of material, a processing container for collecting washed and harvested cells from the separation chamber, and a selection column connected to the fluid circuit for further cell affinity selection processing of the cells from the process container.

[0012] In a fifth aspect, modified cells for use in the treatment of a disease, ailment, or medical condition of a patient are provided wherein the modified cells are produced using a method that includes obtaining a biological fluid processing system comprises afluid processor that operates on a disposable fluid circuit that is connectable to a source container having a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a processing container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit; conducting a pre-processing state further comprising: selecting a procedure protocol, conducting procedure setup, installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor a plurality of containers; running a fluid circuit solution prime including the separation chamber; attaching to the fluid processor the source container; and running a fluid circuit source prime; conducting a processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid; rinsing the source container; diluting the harvested cells; collection of the harvested cells in the processing container; draining solution from the selection column; adding antibodies to the selection column; transferring the harvested cells from the processing container into the selection column; transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste; diluting the material in the selection column; repriming the separation chamber for processing the material in the selection column; target fraction loading from the selection column into the separation chamber; washing and harvesting cells from the selection column; harvesting selected cells as a final product in at least one final container; and conducting a post-processing state further comprising; sealing the at least one final container and removing the final product; and removing the disposable fluid circuit.

[0013] In a sixth aspect, modified cells for use in the treatment of a disease, ailment, or medical condition of a patient are provided wherein the modified cells are produced using a method of operating a biological fluid processing system for cell affinity selection, wherein the system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container filled with a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a process container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit. The method includespre-processing, processing, and post-processing states; the pre-processing state further comprising: selecting a procedure protocol; procedure setup; installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor at least a first solution container comprising a separation / processing buffer, a second solution container comprising a resuspension buffer and a third solution container comprising a releasing buffer; running a fluid circuit solution prime; attaching to the fluid processor the source container; and running a fluid circuit source prime. The processing state further comprises processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid, rinsing of the source container, dilution of the harvested cells, collecting the harvested cells in the processing container, draining buffer solution from the selection column, diluting the material in the selection column via adding antibodies to the selection column, incubating with the antibodies in the selection column, transferring the harvested cells from the processing container into the selection column, transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste, diluting the material in the selection column via transferring biotin into the selection column, incubating, repriming the separation chamber for processing material in the selection column, loading the target fraction from the selection column into the separation chamber, washing and harvesting the cells processed in the separation chamber, rinsing the source container, harvesting selected cells as a final product in at least one final container, and diluting of the harvested cells in the final container; and the post-processing state further comprising: sealing the at least one final container and removing the final product, and removing the disposable fluid circuit.DESCRIPTION OF THE DRAWINGS

[0014] Fig. 1 is a perspective view of an embodiment of a fluid handling system for processing biological fluids and accomplishing affinity cell selection.

[0015] Fig. 2 is a block diagram of a controller used in conjunction with the other elements of the system of Fig. 1 .

[0016] Figs. 3A-3C are flow charts showing detailed steps of a method of operating the system of Fig. 1 , and the system incorporated therein.DETAILED DESCRIPTION

[0017] A more detailed description of the devices and methods that utilize the devices in accordance with the present disclosure is set forth below. It should be understood that the description below of specific devices and methods is intended to be exemplary, and not exhaustive of all possible variations or applications. Thus, the scope of the disclosure is not intended to be limiting, and should be understood to encompass variations or embodiments that would occur to persons of ordinary skill in the art.

[0018] The example fluid processing system 10 of Fig. 1 may include many attributes and structures found in the systems shown in US Patent No. 11 ,478,755, which reference is owned by Applicant, Fenwal, Inc., and is incorporated herein in its entirety.

[0019] Turning to Fig. 1 , an embodiment of a fluid handling system 10 is shown for processing biological fluids, which features affinity cell selection. The system 10 includes a fluid processor 12 connectable to a source container 14 filled with a biological fluid, and is configured to separate the biological fluid from the source container 14 into at least two streams of material. “Biological fluid” includes without limitation blood and blood components, and “cell” or “biological cell” include without limitation blood cells, such as red cells, white cells, and platelets. The processor 12 is connected to a further fluid processing container 16, which may be flexible or rigid walled. The processing container 16 initially may be empty and rests upon a tray or floor 18 of a housing 20. The housing 20 may be open or may include a door or cover, providing a closed state. The tray 18 permits holding, mixing via shaking and the like, and may include a clip to help retain the position of the container 16 on the tray 18. The container 16 also is located along a fluid pathway 22.

[0020] The processor 12 includes a disposable fluid circuit (also referred to as a set or kit) 24 used in combination with a reusable processing machine, or hardware 26. The fluid circuit 24 may be a "closed" system or circuit, in which the interior of thesystem, e.g., the flow paths, containers, etc., are not exposed or "opened" to the outside environment. The fluid circuit 24 may be referred to as closed even where additional containers are attached to the fluid circuit 24, for example before or during a procedure. The fluid circuit 24 includes the fluid pathway 22.

[0021] A control unit (or controller) 28 (see Fig. 2) is coupled to the processor 12, including the fluid circuit 24 and hardware 26. The controller 28 is configured to operate the processor 12, and hardware 26 according to a procedure or process to have the system 10 produce or generate a product. For example, the system 10 may be comprised of a fluid handling system paired with a cell selection column 30. The cell selection column 30 may be integrated into or added to the fluid circuit 24. As further discussed herein, the system 10 is capable of preparing apheresis-collected (or other heterogeneous cell populations) material for cell selection by automated washing, incubation, and volume / concentration adjusting of the cellular material. The system 10 also may facilitate the removal of target / non-target cells fractions by adding selection reagents to the cell selection column 30 and by subsequent washing / resuspension of the target / non-target fraction at desired cell concentrations for downstream use. The target cells initially may be separated from a biological fluid by the processor 12.

[0022] As seen in Fig. 1 , the disposable fluid circuit 24 is connectable to the source container 14 of fluid, in particular biological fluid. In this example, the disposable fluid circuit 24 includes a separation chamber 31 , illustrated as a spinning membrane separator, which is used to process the fluid received from the source container 14. The flow of fluid from the source container 14, through the spinning membrane separator 31 , and to the one or more containers, such as the processing container 16, is achieved through the use of first and second syringes 32, 34, which are in fluid communication with the source container 14, the spinning membrane separator (or spinning membrane for short) 31 , the processing container 16 and the cell selection column 30. The syringes 32, 34 also may be in fluid communication with a number of other containers 36, 38, 40.

[0023] For example, container 36 is initially empty and is available to accept waste generated within the processing; container 38 may contain a processing or resuspension buffer or agent; and container 40 may contain a releasing buffer or agent,such as a biotin releasing agent. Selection column 30 may initially contain a matrix or resin material and may contain a storage buffer, if necessary to maintain wetting of such material.

[0024] Within the fluid circuit 24, the flow of fluid between the containers 14, 16, 36, 38, 40, the spinning membrane 31 , the syringes 32, 34 and the cell selection column 30 is controlled using a flow control cassette 42, which may be connected to each of the foregoing by tubing (or “lines”). In addition, the cassette 42 may include internal flow paths that are defined in part by a plurality of separate channels or passages, which in turn may be contained within and may be defined by the structure (e.g., housing) of the cassette 42. The channels may be connected at a plurality of selectable junctions, which may control the flow of fluid from one channel to another. These selectable junctions also may be referred to as valves, valve stations, or clamps, because the selectable junctions provide controlled access between the channels. The cassette 42 also may include sensor stations, by which sensors may be associated with the flow paths within the cassette 42 to determine characteristics of the flow therein, such as pressure. Preferably, the length of each of the lines and channels is kept as short as possible to further minimize the internal volume of the fluid circuit 24.

[0025] As illustrated in Fig. 1 , the separation chamber or spinning membrane 31 and the syringes 32, 34 may be integrally formed as part of (i.e., as one piece with) the cassette 42, to further reduce the tubing volume associated with the fluid circuit or kit 24. According to other embodiments, the spinning membrane 31 and / or the syringes 32, 34 may be attached to the remainder of the fluid circuit 24 at the time of use, as may be the case with one or more of the containers 14, 16, 36, 38, 40. Again, as illustrated in Fig. 1 , the container 36 and container 16 may be integrally formed with the cassette 42.

[0026] As seen in Fig. 1 , the reusable hardware component (or reusable hardware for short) 26 includes a drive 43 for the spinning membrane separator 31 , a syringe pump 44, 46 for each respective syringe 32, 34, and a control cassette interface 48 that is associated with the flow control cassette 42 when the fluid circuit 24 is disposed on the hardware 26 (e.g., is mounted on the hardware 26). The cassette interface 48 includes actuators and sensors that are associated with the clamps and sensor stationsof the flow control cassette 42 and are configured to operate the clamps or sense characteristics of the fluids passing therethrough, respectively.

[0027] The reusable hardware 26 is coupled to the controller 28 which is configured to control operation of the system 10, for example using a method of operation as is explained below in relation to the flow charts in Figs. 3A-3C. As seen in Fig. 2, the controller 28 may include a microprocessor 50, which may include multiple physical and / or virtual processors. According to other embodiments, the controller 28 may include one or more electrical circuits designed to carry out the actions described herein. Thus, the controller 28 may include a microprocessor 50 and other circuits or circuitry. In addition, the controller 28 may include at least one memory 52. The instructions by which the microprocessor 50 is programmed may be stored on the at least one memory 52 associated with the microprocessor 50. The at least one memory 52 may include one or more tangible non-transitory computer readable memories, having computer executable instructions stored thereon, which when executed by the microprocessor 50, may cause the microprocessor 50 to carry out one or more actions as described herein.

[0028] As mentioned above, the controller 28 may be coupled (i.e., directly or indirectly connected) to the equipment of the reusable hardware 26, such as the spinning membrane drive 43, the first syringe pump 44, the second syringe pump 46, and the cassette interface 48. The controller 28 may operate each of these devices, each of which may be an assembly of other devices or equipment, to cause fluids to flow through the fluid circuit 24 associated with the hardware 26, for example to cause fluid to flow from the source container 14, through the spinning membrane 31 , and eventually into the processing container 16.

[0029] For instance, the controller 28 may be programmed to perform a process or procedure according to a protocol, such as to wash particular cells contained in the biological fluid within the source container 14, before they are directed to the processing container 16. The controller 28 may be programmed to perform other actions as well, such as to test the fluid circuit 24, to prime the fluid circuit 24, to rinse parts of the circuit 24 after the wash has been performed, and to add other components to the cell-containing fluid or complete other steps before that fluid is distributed to the processing container 16.

[0030] As also illustrated in Fig. 2, the controller 28 may be coupled to one or more of the structures or devices described above, for example to receive information (e.g., in the form of signals) from these structures or to provide commands (e.g., in the form of signals) to these structures to control the operation of the structures. As illustrated, the controller 28 may be coupled to at least one input device 54 to receive information from that device and to at least one output device 56 to provide information to that device. The at least one input device 54 may include a number of different devices according to the embodiments described herein. For example, the input device 54 may include a keyboard or keypad by which a user may provide information and / or instructions to the controller 28.

[0031] Alternatively, the input device 54 may be a touch screen, such as may be used in conjunction with an output device 56 in the form of a video display. For instance, the illustrated embodiment in Fig. 1 includes such a touch sensitive display screen 58 mounted to a front panel of a housing 60 of the fluid processor 12 of the system 10. The input device also may include a reader or scanner, such as a barcode reader, scanner or RFID reader 62. According to still other embodiments, the input device 54 may be in the form of computer equipment that permits the cell processing system 10 including the controller 28 to communicate (whether via wires, cables, etc., or wirelessly) with other processing systems over a local network, or with other cell processing systems or other computer equipment (e.g., a server) over local networks, wide area networks, or the Internet. According to such an embodiment, the input device may alternatively include an internal transmitter / receiver device.

[0032] The controller 28 may also be coupled to alternative devices, such as dispenser for automated injection, for example, of antibodies into the cell selection column 30. As a further example, the controller 28 may be coupled to a means to agitate fluid in the processing container 16. To permit the fluid in the processing container 16 to be agitated, the housing 20 may be mounted on a shaft 64 that depends from a motor located in the housing 60 of the processor 12. The motor may cause the shaft 64 to rotate in opposite directions about an axis of the shaft. Thus, the fluid in theprocessing container 16 resting on the tray 18 of the housing 20 may be agitated in an oscillatory fashion.

[0033] Mounting of the housing 20 on the shaft 64 also may permit the processing container 16 to be disposed at an angle relative to horizontal, instead of having the container disposed such that the container is level. For example, the tray 18 of the housing 20 may be oriented at a 30 degree angle relative to horizontal to influence the fluid in the processing container 16. Of course, this could be achieved in a standalone version of the housing 20 as well, either by providing a mechanism (such as a pivot) that permits the housing 20 (or surface of tray 18) to be disposed at a variable angle or a mechanism (such as an inclined plane) that permits orientation at a particular angle.

[0034] The system 10 may be automated or function automatically. This is to say that the system 10 (and in particular controller 28) may be programmed to carry out the processing steps of a processing method without requiring intervening substantial operator / user involvement. Of course, even in the automated system of the present disclosure, it will be understood that user activity may be involved, including the loading of the disposable fluid circuits and entering processing parameters. Additional manual steps may be required, as well, such as connecting additional fluid sources or containers, etc., and there may be opportunities for reconfiguration of certain steps in a process. However, the reusable apparatus of system 10 can process biological fluids through the disposable circuit(s) described below without substantial user intervention.

[0035] A detailed description of methods of operation of the system begins, in general terms, with the methods including multiple steps within phases or states of Pre- Processing 100, Processing 200, and Post-Processing 300. Flow charts representing some alternatives for the steps in these states can be seen in the figures, such as preprocessing 100 in Fig. 3A, processing 200 in Fig. 3B and post-processing 300 in Fig. 3C.

[0036] The user may first activate (e.g., switch on) the hardware 26. The hardware 26 in conjunction with the controller 28 may conduct self-calibration checks, including checking the pumps 44, 46, and other components. Similar self-calibration checks may be performed relative to particular devices, such input or output devices, or when the user activates the hardware 26.

[0037] As shown in Fig. 3A, Pre-Processing 100 starts at block 102 with Protocol Selection. In this step, the fluid processing system 10 displays a list of protocols from which to choose to run a cell processing procedure, such as on the touch screen 58. The related protocol parameters for this step may include a respective Protocol ID and Protocol Description.

[0038] Next, at block 104 the method includes Procedure Setup. During Procedure Setup for a selected protocol, the system controller 28 prompts the user to enter information, which for example may include identification, consumables, and source composition that are pertinent to the specific selected procedure to be executed. Such information may be entered via the input device 54. For this step 104, the related protocol parameters may more specifically include pertinent information, depending on the procedure, for example, including: a required Procedure ID, User ID and Source ID, as well, for example identifying the Target Cell, Cell Component, Component Retention Flag (for Secondary Wash and Post-Selection Wash), Primary Set Reference Number, Primary Set Confirm Reference Configured, Solution 1 Name, Solution 1 Reference Number, Solution 2 Name, Solution 2 Reference Number, Solution 3 Name, Solution 3 Reference Number, and Final Product Bag(s) Max Fill Volume (mL). Also, the controller may prompt the user to enter or modify process parameters using the input device 54 relating to functions of the hardware 26, including by way of example and not by way of limitation the amount of cell suspension to be processed, the number of cycles to perform, etc.

[0039] Pre-Processing 100 next includes at block 106 a step to Install Set. This involves the controller 28 of the fluid processing system 10 prompting the user to install the disposable fluid circuit 24 (also referred to as the disposable kit, kit or cassette) onto the hardware 26 of the processor 12 of the fluid processing system 10. During this step, the fluid processing system 10 will load and seal the fluid circuit 24 into place. Once the kit 24 is installed, at block 108 Installation Checks are performed. In this step, the system 10 checks various parts of the disposable kit 24 for correct installation and confirms the kit integrity.

[0040] In the illustrated example, Pre-Processing 100 continues at block 110 with Attach Solution 1 , wherein the system 10 prompts the user to attach Solution 1 to thedisposable kit 24. Solution 1 may be a separation / processing buffer or agent, which may be held in a container, such as container 38. The related protocol parameter for this step may include the Solution 1 connected volume. When connecting the solutions, the connections, such as a connection of a tubing to a container, may be made via a spike connector, sterile connection, or other suitable means of connection.

[0041] This is followed at block 112 by the step of Attach Solution 2, wherein the system 10 prompts the user to attach Solution 2 to the disposable kit 24. Solution 2 may be for example a resuspension buffer or agent, such as saline solution, which may be held in a container, such as container 40. The related protocol parameter for this step may include the Solution 2 connected volume.

[0042] The next step shown in Fig. 3A is at block 114 with the step Attach Solution 3. In this step, the system 10 prompts the user to attach Solution 3 to the disposable kit 24. Solution 3 may be, for example, a releasing buffer or agent, one example of which may be a biotin containing solution, which may be held in a container, such as container 40. The related protocol parameters for this step may include the Solution 3 connection point and Solution 3 connected volume. The system 10 also may be configured in an alternative manner to attach Solution 3 prior to another step that would occur midprocedure. An optional step may include addition of another solution, depending on the desired processing. If applicable, the system 10 may prompt the user to attach a container of such additional solution to the disposable kit 24.

[0043] Once the user has connected the appropriate Solutions, the Pre-Processing 100 continues at block 116 with Solution Prime. The controller 28 of the fluid processing system 10 primes various parts of the fluid circuit or disposable kit 24 to (1 ) check for correct installation of the tubing lines and (2) clear tubing lines and the spinning membrane separator 31 of air. In an example embodiment, the fluid circuit 24 may be primed with saline, although other biocompatible aqueous solutions may be used.

[0044] The next step at block 118 is Attach Source, wherein the controller 28 of the system 10 prompts the user to attach the Source product to the kit 24. The source product may be biological fluid / cells, which may have been recently obtained via apheresis collection, leukapheresis, or other heterogenous cell populations, refrigeratedovernight or previously cryopreserved material, etc., and may be held for example in source container 14.

[0045] This is followed at block 120 by Source Prime, wherein the system primes the tubing line leading to the Source product in container 14 to clear the tubing line of air. This state optionally also can be used to pre-dilute the Source product, if desired. The related protocol parameters for this step may include the Source Prime volume (mL) and Source Prime flow rate (mL / min). At this stage, the method may include block 122 for a Source Prime Pause, wherein the system 10 may be configured to pause at the completion of Source Prime step 120 to agitate or mix the Source product container 14 before transitioning to the next state. As such, the related protocol parameters for this step may include Post Prime Pause Configured and Post-Prime Pause Text.

[0046] Having completed the steps of Pre-Processing 100, illustrated in Fig. 3A, the method then enters the state of Processing 200, as seen in Fig. 3B. Fig. 3B includes circled, numbered notes 1 to 6, which correspond to a key shown in Fig. 3C and which related to which processing path will be followed, such as when to perform a repeated set of operations or to move on to another stage of the processing.

[0047] Processing 200 starts at block 202 with Source Loading (for a Primary Wash cycle). In this step 202, Source product is drawn into the spinning membrane separator 31 , such as through a port on the Source product container 14 and via tubing and use of the first syringe 32, associated with syringe pump 44. Thus, the controller 28 causes the biological fluid / cells to be transferred from Source container 14 through the fluid circuit or kit 24 to its spinning membrane separator 31 via operation of one or more syringe pumps 44, 46. In a similar fashion, the wash medium may be delivered from its container 38 through the fluid circuit 24 to the spinning membrane separator 31 .

[0048] At block 210, during the Primary Wash, the cells within the Source product accumulate within the annular space of the spinning membrane separator 31 until either (1 ) the Source container is empty (identified in Fig. 3B by circled key 2: Source Container Empty) or (2) the number of cells within the spinning membrane separator 31 reach a pre-configured capacity (identified in Fig. 3B by circled key 1 : Maximum Annular PCV% Reached). As cells are loaded into the separation chamber or spinning membrane 31 , supernatant (filtrate) crosses the membrane and is drawn into firstsyringe 32. It will be appreciated that the biological cells may be collected or harvested at block 212 Harvest in an in-process container 16, while supernatant is separated and removed to a waste container 36. The related protocol parameters for this step may include spinning membrane separator Loading Revolution Rate (RPM), Source Inlet Flow Rate (mL / min), and Maximum Annular packed-cell volume (PCV) (%).

[0049] The Processing 200 continues with further Source Loading 202 and Wash 210 (for Primary Wash), wherein with the cells suspended within the annular space of the spinning membrane separator 31 , Solution 1 is drawn into the spinner of the separator 31 using the first syringe 32. Residual supernatant and wash buffer cross the spinning membrane 31 and are drawn into the first syringe 32. The related protocol parameters for this step may include Primary Spinner Wash Configured, Primary Wash Solution, Primary Wash Spinner Wash Volume (mL) and Primary Wash Spinner Wash Flow Rate (mL / min). It also will be appreciated that this step may be repeated with an additional wash solution. For example, when processing fresh apheresis material for selection, platelets become activated with washing Solution 2. In that instance, the suspension is first washed with Solution 1 to remove platelets to the filtrate, which may then be immediately followed by a wash with Solution 2 to resuspend the cells in the media required for the subsequent steps of processing.

[0050] The step Wash 210 is followed at block 212 by Harvest (for Primary Wash), wherein the further cells within the annular space of the spinner of the spinning membrane separator 31 are drawn out of the spinner with Solution 1 using the second syringe 34 and are transferred to the in-process container, such as container 16 in the example shown, or to an alternative processing container. If the Source container 14 is not empty, the system 10 will return to the Source Loading (for Primary Wash) state (as identified in Fig. 3B by circle key 3: Source Container Not Empty).

[0051] It will be appreciated that these steps of Source Loading 202, Wash 210 and Harvest 212 may be repeated until, for example, the Source container 14 is empty. The related protocol parameters for the step Harvest 212 (for Primary Wash) may include Harvest Volume (mL), and Harvest Flow Rate (mL / min).

[0052] This is followed at block 204 by Source Rinse (for Primary Wash). The system 10 then returns to a Source Loading state, at block 208, to process (Wash andHarvest) the rinse solution with the residual cells. However, the system 10 can be configured to include at block 206 a Source Rinse Pause after Solution 1 is transferred to the Source container 14, to allow the user to mix or agitate the rinse solution in the Source container 14 to enhance the ability to capture any residual cells trapped in the Source container 14. The text displayed on screen 58 may be configurable to permit the addition of this step. The related protocol parameters for this step may include Source Rinse Pause Configured and Source Rinse Pause Text. If the Source Rinse Pause is utilized, then system 10 thereafter moves to the block 208 step of Source Loading which is to process (Wash and Harvest) the rinse solution with the residual cells.

[0053] If the Source container 14 finally is empty, the system 10 will transition at block 214 to a step Dilution 1 . In the step Dilution 1 , the system 10 dilutes the cell suspension in the processing container 16 immediately after harvesting the cells from the spinning membrane separator 31 with Solution 1 . The related protocol parameters for step Dilution 1 may include Dilution 1 Volume (mL), and Dilution 1 Flow Rate (mL / min).

[0054] Following step Dilution 1 at block 214, the process may proceed to the next step at block 216 which is Dilution 1 Pause. In Dilution 1 Pause, the system 10 may be configured to pause at the completion of the Dilution 1 state. The related protocol parameters for this Dilution 1 Pause may include Dilution 1 Pause Configured, Dilution 1 Pause Text, Dilution 1 Sample Volume (mL) and Antibody Volume (mL).

[0055] The Dilution 1 Pause at 216 is followed at block 218 by Drain Storage Buffer from Selection Column. Thus, in this particular example, a buffer solution initially held in the selection column 30 may be drained, leaving the matrix resin material. Note that, if using a buffer solution not requiring pre-wetting, the procedure may skip this step. This may be followed at block 220 by Wash / Rinse Selection Column to prepare the selection column 30 for further processing.

[0056] The next step, at block 222, is Dilution 2 (Transfer Ab into Selection Column). This is the point in the procedure shown in Fig. 3B, at which the user may add antibody directly to the selection column 30. For example, the user may manually inject an antibody fragment (FAB) solution into the section column 30 in a sterile manner.According to other embodiments the antibody may be introduced automatically into the selection column 30.

[0057] This is followed at block 224 by Incubation. In this example, the Incubation is antibody incubation wherein the matrix material in the selection column 30 incubates with the added antibodies. The duration of the incubation are configurable by the user via the at least one input device 54. This is followed at block 226 by Transfer Washed Cells to Selection Column, where the cell suspension that has been collected in the processing container 16 is transferred to the selection column 30.

[0058] This is followed at block 228 by a further Incubation step. The duration of the incubation is configurable by the user, such as via the input device 54. The related protocol parameters may include Incubation Time.

[0059] Following the Incubation at block 228, the procedure may proceed to block 240 for a Target Fraction Loading from Selection Column step (as identified in Fig. 3B by circle key 6: Negative Selection Only), or the process may proceed to the next step at block 230, which is Transfer Non-Target Fraction to Waste.

[0060] The Processing 200 continues at block 230 for the step of Transfer Negative Fraction To Waste. In this step 230, the unbound cells that remain in suspension are removed from the selection column 30 via the second syringe 34. The removed cells are then transferred to the waste container 36. The number of negative selections is configurable by the user by using the input device 54. There may be no additional negative selection configured (as identified in Fig. 3B by circle key 5: No Additional Selection). Alternatively, if an additional negative selection is configured, the system to advances to block 232 for Dilution 3, and again dilutes and re-suspends the cells remaining in the selection column 30 with Solution 1 , re-incubates, and repeats the transfer of the negative fraction to waste. The related protocol parameters at block 230 may include Number of Negative Selections. As noted, Dilution 3 is provided at block 232. The related protocol parameters may include Dilution 3 Flow Rate (mL / min), Dilution 3 Solution, and Dilution 3 Volume (mL).

[0061] At block 234, Dilution 4 (Transfer Release Agent into Selection Column), a release agent, such as biotin is transferred into the selection column 30. This is followed at block 236 by Incubation. This incubation may provide, for example, releasebuffer incubation wherein the cell suspension in the selection column 30 incubates with Solution 3. The duration of the incubation is configurable by the user via the at least one input device 54. The related protocol parameters may include Incubation Time. Use of such an incubation state may be desired, depending on the release time associated with the release agent, such as with biotin affinity in the selection column 30.

[0062] The system 10 then advances to block 238 for Spinner Re-Prime. In this step, the system 10 re-primes segments of the fluid circuit or disposable kit 24, such as with a Solution 2 and use of the second syringe 34 to clear air from the fluid circuit 24. The related protocol parameters for connecting Solution 2 may include Solution 2 Connected Volume, while the related protocol parameters for Spinner Re-Prime may include Post-Selection Prime Volume (mL), and Post-Selection Source Prime Flow Rate (mL / min).

[0063] The system 10 then advances to block 240 for Target Fraction Loading from Selection Column. The cell suspension is drawn into the spinner of the spinning membrane separator 30 through the source port using the first syringe 32. The cells accumulate within the annular space of the spinner until either (1 ) the selection column 30 is empty (as identified in Fig. 3B by circle key 2: Source Container Empty), in which case the next step is at block 242 Selection Column Rinse (discussed below), or (2) the number of cells within the spinning membrane 31 reach a pre-configured capacity (as identified in Fig. 3B by circle key 1 : Maximum Annular PCV% Reached), in which case the next step is at block 248, Wash. As cells are loaded into the spinner of the spinning membrane separator 31 , supernatant (filtrate) crosses the membrane and is drawn into first syringe 32. The related protocol parameters for such a post-selection Target Fraction Loading from Selection Column may include Post-Selection Spinner Loading Revolution Rate (RPM), Post-Selection Source Inlet Flow Rate (mL / min), and PostSelection Maximum Annular PCV (5%).

[0064] When the selection column 31 is empty (as identified in Fig. 3B by circle key 2: Source Container Empty) the method proceeds to block 242 for Selection Column Rinse. This is a post-selection Selection Column Rinse, wherein when the selection column 31 is empty, Solution 2 is transferred at block 238 to the selection column 30 to recover any residual cells. The system returns to the post-selection Target FractionLoading state to process the rinse solution with the residual cells. The related protocol parameters for this Source Rinse step may include Post-Selection Source Rinse Configured, and Post-Selection Source Rinse Volume (mL).

[0065] At block 244 the system 10 reaches a Selection Column Rinse Pause. This is a post-selection Selection Column Rinse Pause wherein after Solution 2 is transferred to the selection column 31 , the system 10 may be configured to pause to allow the user to mix or agitate the rinse solution in the selection column 31 to capture any trapped residual cells. The text displayed on the display screen 58 is configurable, such as via the input device 54. The related protocol parameters for this step may include PostSelection Selection Column Rinse Pause Configured, and Post-Selection Selection Column Rinse Pause Text. This continues at block 246 Target Fraction Loading from Selection Column, in preparation for a further Wash at block 248.

[0066] The next step is at block 248 Wash (New Media), whether proceeding from block 240 due to the number of cells within the spinner reaching a pre-configured capacity (as identified in Fig. 3B by circle key 1 : Maximum Annular PCV% Reached), or progressing from the further Rinse steps and Target Fraction Loading from Selection Column at block 246. With the cells suspended within the annular space of the spinner of the spinning membrane separator 31 , Solution 2 is drawn into the separation chamber 31 using the first syringe 32. Residual supernatant and wash buffer cross the membrane and are drawn into the first syringe 32. The related protocol parameters for this step may include Post-Selection Spinner Wash Configured, Post-Selection Wash Volume (mL), and Post-Selection Wash Flow Rate (mL / min).

[0067] The Processing 200 continues at block 250 for Harvest (to Final Container(s)). In this Harvest step, the cells within the annular space of the spinner of the spinning membrane separator 31 are drawn out of the spinner with Solution 2 using the second syringe 34 and transferred to the final product container 66, which would be a user-connected container (either bag, culture vessel, or other container) connected to fluid pathway 22. If the selection column 30 is not empty (as identified in Fig. 3B by circle key 3: Source Container Not Empty), the system 10 will return to the Target Fraction Loading from Selection Column step at block 240. If the selection column 30 is empty, the system 10 will transition to the Final Dilution (New Media) state at block 252.The related protocol parameters for this step may include Harvest Volume (mL), and Harvest Flow Rate (mL / min).

[0068] In the further Final Dilution (New Media) step at block 252, the system dilutes the cell suspension in the final product container 66 connected to fluid pathway 22 immediately after harvesting the cells from the spinner of the spinning membrane 31 with Solution 2 to achieve the target final product volume or target final concentration. The related protocol parameters for this step may include Final Dilution Flow Rate (mL / min), Final Dilution Entry Method, Final Dilution Concentration, Final Dilution Volume (mL), and Final Dilution Air Chase Volume (mL).

[0069] The system 10 then transitions to Post-Processing 300, shown in Fig. 3C, which includes at block 302 Seal / Remove Final Product. At this step, the system 10 prompts to seal and remove the final product container 66 attached to fluid pathway 22. This may be followed at block 304 by a Procedure Summary step, wherein the system displays a summary of the procedure, such as on the screen display 58. Finally, at block 306 Remove Set, the system 10 prompts the user to remove the disposable fluid circuit or kit 24 from the fluid processing hardware 26.

[0070] According to certain embodiments, as shown in Fig. 1 , the seal and removal of the final product may be in one product container 66. Alternatively, as shown in Fig.1 , the system also may provide further tubing and containers to aliquot the product of the cell selection into a plurality of smaller product containers 66A for having a volume appropriate for administration or delivery to a patient. Such product containers also may be configured as delivery containers, such as syringes.

[0071] Thus, improved fluid handling systems for processing biological fluids and accomplishing cell selection are discussed herein. The description provided above, and the other aspects provided below, are intended for illustrative purposes, and are not intended to limit the scope of the disclosure to any particular method, system, apparatus, or device described herein.

[0072] It will be appreciated that operating the biological fluid operating system 10 includes processing or modifying the target cells, cell separation, concentration and / or washing, cell modification including but not limited to genetic modification, or other suitable forms of cell processing. As used herein, the phrase “cell modification” or“modification” or “modified” refers to a process in which target cells are isolated from other cells and then mixed with a modification solution, agent, or utilized with the vectors described below.

[0073] In one embodiment, modified cells produced by the blood processing system include Chimeric Antigen Receptor T-Cells (CAR-T) cells. These CAR-T cells may be used in several applications and therapeutic treatments of various diseases, ailments, or conditions. For example, the CAR-T cells may be used to treat solid tumors such as anal / rectal, epithelial, ovarian, breast, fallopian tube, endometrial, pancreatic, colorectal, lung, and / or gastrointestinal tumors. The CAR-T cells may also be useful in therapeutically treating melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic lymphoma, non-Hodgkin lymphoma, diffuse large b-cell lymphoma, small-cell lung cancer, neuroblastoma, glioblastoma, prostate cancer, hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, and rheumatoid arthritis. The CAR-T cells may also be useful for renal transplantation patients. Accordingly, the disclosure provides a modified CAR-T cell produced by the blood processing system for use in the treatment of a disease or condition selected from the group consisting of solid tumors such as anal / rectal, epithelial, ovarian, breast, fallopian tube, endometrial, pancreatic, colorectal, lung, and / or gastrointestinal tumors; melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic lymphoma, non-Hodgkin lymphoma, diffuse large b-cell lymphoma, small-cell lung cancer, neuroblastoma, glioblastoma, prostate cancer, hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, rheumatoid arthritis, and impairments related to renal transplantation.

[0074] In producing the CAR-T cells, different modification strategies may be used in the blood processing system, with the various methods being practiced in the preparation of CAR-T cells for any one of the preceding applications or therapeutic treatments. Vectors such as mRNA, shRNA, siRNA, saRNA, SeekRNA, polymers, proteins and peptides, antibodies, viruses, including but not limited to lentivirus (LV),Adeno Associated Virus (AAV), labelling molecules, small molecules, Virus-like particles (VLP), transposon / transposase (sleeping beauty, TcBuster, PiggyBac), transcription activator-like effector nuclease (TALEN), zinc finger nucleases (ZFN), base editors, prime editors, programmable addition via site-specific targeting elements (PASTE), CRISPR / Cas, and CRISPR RNPs may be utilized in the blood processing system (specifically the cargo delivery module) to create the CAR-T cells. Accordingly, the disclosure provides in certain embodiments modified CAR-T cells produced by use of the aforementioned vectors for use in the treatment of a disease, ailment, or condition as shown in Tables 1 and 2:Table 1Table 2

[0075] Embodiments 1 to 59 listed in Tables 1 and 2 do not only apply to modified CAR-T cells, but also to any other modified cell types that can be produced by the biological fluid processing system.

[0076] Accordingly, a “modified cell” as used herein is a cell that has been produced by the biological fluid processing system by use of the vectors of embodiments 35 to 59 (Table 2). Said modified cell is further provided for use in the treatment of a disease, ailment, or condition according to embodiments 1 to 34 (Table 1 ).

[0077] In an embodiment, the modified cells are modified Chimeric Antigen Receptor Natural Killer (CAR-NK) cells.

[0078] In a further embodiment, the modified cells are modified Chimeric Antigen Receptor Monocytes (CAR-M) cells.

[0079] In another embodiment, the modified cells are modified Engineered T-Cell Receptor (TCR) cells.

[0080] In a further embodiment, the modified cells are modified Engineered B-Cells.

[0081] In another embodiment, the modified cells are modified Tumor Infiltrating Lymphocytes (TIL) cells.

[0082] In a further embodiment, the modified cells are modified Induced Pluripotent Stem (iPSC) Cells.

[0083] In another embodiment, the modified cells are modified Invariant Natural Killer T (iNKT) cells.

[0084] In a further embodiment, the modified cells are modified Mesenchymal Stem (MSC) cells.

[0085] In another embodiment, the modified cells are modified Dendritic Cells.

[0086] In a further embodiment, the modified cells are modified Hematopoietic Stem Cells / Stem (CD34+) cells.

[0087] The modified cells can be produced and administered in a variety of environments by the blood processing system, with the modified cells being produced using any one of the preceding vectors and being administered in any one of the preceding applications or therapeutic treatments. By way of example, the blood processing system can produce and / or administer modified cells within a patient treatment room / facility (inpatient or outpatient). The blood processing system can produce and / or administer modified cells within a cell processing lab. The cell processing lab may be at a hospital facility, at a non-hospital facility, or a commercial production facility (centralized or decentralized). The blood processing system can produce and / or administer modified cells within a sterile manufacturing suite. The sterile manufacturing suite may be at a hospital facility, at a non-hospital facility, at an academic facility, or at commercial production facility (centralized or decentralized).

[0088] While the modified cells produced by this system are described as being used to treat particular conditions and patient groups, it should be understood that they may be applied to other conditions and / or patient groups, including sub-groups of a described patient group (i.e., patients having the same characteristics that characterize a particular patient group, but additional characteristics that are not shared by all patients of that patient group), larger patient groups encompassing a described patient group (i.e., a patient group having broadly defined characteristics that include the characteristics that characterize a particular patient group) and entirely different patientgroups (i.e., patients having characteristics that exclude them from a particular patient group). The modified cells may also be applied in various dosages, administrative regimes, and routes of administration without departing from the scope of the present disclosure.

[0089] It should be understood that the embodiments disclosed herein may be combined with each other in any imaginable combination.Other Aspects

[0090] Aspect 1 . A biological fluid processing system for cell affinity selection, including a fluid processor having reusable hardware that operates on a disposable fluid circuit, the fluid circuit being connectable to a source container and having a biological fluid, a separation chamber configured to separate the biological fluid from the source container into at least two volumes of material, a processing container for collecting washed and harvested cells from the separation chamber, and a selection column connected to the fluid circuit for further cell affinity selection processing of the cells from the process container.

[0091] Aspect 2. The system of aspect 1 , wherein the fluid processor further comprises a controller configured to operate the reusable hardware while the disposable fluid circuit is installed on the reusable hardware and is connected to the source container, the processing container and the selection column.

[0092] Aspect 3. The system of aspect 2, wherein the controller further comprises at least a micro-processor and memory.

[0093] Aspect 4. The system of any one of aspects 1 -3, wherein the fluid processor further comprises an input device and an output device connected to the controller.

[0094] Aspect 5. The system of any one of aspects 1 -4, wherein the reusable hardware of the fluid processor further comprises a drive for a spinning membrane separator in the separation chamber.

[0095] Aspect 6. The system of any one of aspects 1 -5, wherein the reusable hardware of the fluid processor further comprises pumps and a cassette interface that engage the fluid circuit.

[0096] Aspect 7. The system of any one of aspects 1 -6, wherein the fluid circuit further comprises a single final product container.

[0097] Aspect 8. The system of any one of aspects 1 -7, wherein the fluid circuit further comprises a plurality of final product containers.

[0098] Aspect 9. The system of any one of aspects 1 -8, wherein the selection column initially contains a matrix or resin material.

[0099] Aspect 10. The system of any one of aspects 1 -9, wherein at least three further containers are connected to the fluid circuit, comprising a first solution container comprising a separation / processing buffer, a second solution container comprising a resuspension buffer and a third solution container comprising a releasing buffer.

[0100] Aspect 1 1 . The system of any one of aspects 1 -10, wherein the controller is coupled to a separator drive for a spinning membrane separator of the separation chamber, the at least two syringe pumps and a control cassette interface, the controller configured to selectively operate the separator drive, the syringe pumps and the cassette interface to provide a procedure utilizing the selection column according to a protocol.

[0101] Aspect 12. A method of operating a biological fluid processing system for cell affinity selection, the method comprising: obtaining a biological fluid processing system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container having a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a processing container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit; conducting a pre-processing state further comprising: selecting a procedure protocol, conducting procedure setup, installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor a plurality of containers; running a fluid circuit solution prime including the separation chamber; attaching to the fluid processor the source container; and running a fluid circuit source prime; conducting a processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid;rinsing the source container; diluting the harvested cells; collection of the harvested cells in the processing container; draining solution from the selection column; adding antibodies to the selection column; transferring the harvested cells from the processing container into the selection column; transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste; diluting the material in the selection column; repriming the separation chamber for processing the material in the selection column; target fraction loading from the selection column into the separation chamber; washing and harvesting cells from the selection column; harvesting selected cells as a final product in at least one final container; and conducting a post-processing state further comprising; sealing the at least one final container and removing the final product; and removing the disposable fluid circuit.

[0102] Aspect 13. The method of aspect 12, wherein the processing utilizes only negative selection relative to target cells.

[0103] Aspect 14. The method of aspect 12 or 13, wherein the processor is configured to further perform positive selection incubation and removal of positive fraction relative to target cells.

[0104] Aspect 15. The method of any one of aspects 12-14, wherein the final product is aliquoted into a plurality of smaller containers before removing the fluid circuit.

[0105] Aspect 16. The method of any one of aspects 12-15, wherein the processor transfers a negative fraction to waste from the selection column, followed by dilution of the material in the selection column with a releasing buffer.

[0106] Aspect 17. The method of any one of aspects 12-16, wherein the releasing buffer is biotin.

[0107] Aspect 18. The method of any one of aspects 12-17, wherein the processor further comprises a controller configured to selectively operate the processor to provide a procedure utilizing the separation chamber, processing container and selection column according to a protocol.

[0108] Aspect 19. A method of operating a biological fluid processing system for cell affinity selection, wherein the system comprises a fluid processor that operates on adisposable fluid circuit that is connectable to a source container filled with a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a process container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit, the method comprising: pre-processing, processing, and post-processing states; the pre-processing state further comprising: selecting a procedure protocol; procedure setup; installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor at least a first solution container comprising a separation / processing buffer, a second solution container comprising a resuspension buffer and a third solution container comprising a releasing buffer; running a fluid circuit solution prime; attaching to the fluid processor the source container; and running a fluid circuit source prime; the processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid, rinsing of the source container, dilution of the harvested cells, collecting the harvested cells in the processing container, draining buffer solution from the selection column, diluting the material in the selection column via adding antibodies to the selection column, incubating with the antibodies in the selection column, transferring the harvested cells from the processing container into the selection column, transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste, diluting the material in the selection column via transferring biotin into the selection column, incubating, repriming the separation chamber for processing material in the selection column, loading the target fraction from the selection column into the separation chamber, washing and harvesting the cells processed in the separation chamber, rinsing the source container, harvesting selected cells as a final product in at least one final container, and diluting of the harvested cells in the final container; and the post-processing state further comprising: sealing the at least one final container and removing the final product, and removing the disposable fluid circuit.

[0109] Aspect 20. The method of aspect 19, wherein the fluid processor further comprises a controller configured to selectively operate the fluid processor to provide a procedure utilizing the separation chamber, processing container and selection column according to a protocol.

[0110] Aspect 21 . Modified cells for use in the treatment of a disease, ailment, or medical condition of a patient, characterized in that the modified cells are produced using a biological fluid processing system, including a fluid processor having reusable hardware that operates on a disposable fluid circuit, the fluid circuit being connectable to a source container and having a biological fluid, a separation chamber configured to separate the biological fluid from the source container into at least two volumes of material, a processing container for collecting washed and harvested cells from the separation chamber, and a selection column connected to the fluid circuit for further cell affinity selection processing of the cells from the process container.

[0111] Aspect 22. The modified cells of Aspect 21 wherein the target cells are selected from the group consisting of chimeric antigen receptor T-cells, chimeric antigen receptor natural killer cells, chimeric antigen receptor monocyte cells, engineered T-cell receptors, engineered B-cells, tumor infiltrating lymphocytes, induced pluripotent stem cells, mesenchymal stem cells, dendritic cells, and hematopoietic stem cells.

[0112] Aspect 23. The modified cells of any one of Aspects 21 -22 wherein the target cells are modified using a vector selected from the group consisting of lentivirus, adeno associated virus, virus-like particles, transposon / transposase, transcription activator-like effector nuclease, zinc finger nucleases, mRNA, shRNA, siRNA, SeekRNA, CRISPR / Cas, base editors, prime editors, and programmable addition via site-specific targeting elements.

[0113] Aspect 24. The modified cells of any one of Aspects 21 -23 wherein the modified cells are produced in an environment selected from the group consisting of a patient treatment room of an inpatient facility, a patient treatment room of an outpatient facility, a cell processing lab of a hospital facility, a cell processing lab of a non-hospital facility, a cell processing lab of a centralized commercial production facility, a cell processing lab of a decentralized commercial production facility, a sterile manufacturing suite of a hospital facility, a sterile manufacturing suite of a non-hospital facility, a sterilemanufacturing suite of an academic facility, a sterile manufacturing suite of a centralized commercial production facility, and a sterile manufacturing suite of a decentralized commercial production facility.

[0114] Aspect 25. The modified cells of any one of Aspects 21 -24 wherein said disease, ailment, or medical condition is selected from the group consisting of solid tumors, melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic leukemia, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, small-cell lung cancer, renal transplantation, neuroblastoma, glioblastoma, prostate cancer, hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, and rheumatoid arthritis.

[0115] Aspect 26. Modified cells for use in the treatment of a disease, ailment, or medical condition of a patient, characterized in that the modified cells are produced using a method including obtaining a biological fluid processing system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container having a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a processing container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit; conducting a pre-processing state further comprising: selecting a procedure protocol, conducting procedure setup, installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor a plurality of containers; running a fluid circuit solution prime including the separation chamber; attaching to the fluid processor the source container; and running a fluid circuit source prime; conducting a processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid; rinsing the source container; diluting the harvested cells; collection of the harvested cells in the processing container; draining solution from the selection column; adding antibodies to the selection column; transferring the harvested cells from the processing container into the selection column; transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste; diluting thematerial in the selection column; repriming the separation chamber for processing the material in the selection column; target fraction loading from the selection column into the separation chamber; washing and harvesting cells from the selection column; harvesting selected cells as a final product in at least one final container; and conducting a post-processing state further comprising; sealing the at least one final container and removing the final product; and removing the disposable fluid circuit.

[0116] Aspect 27. The modified cells of Aspect 26 wherein the target cells are selected from the group consisting of chimeric antigen receptor T-cells, chimeric antigen receptor natural killer cells, chimeric antigen receptor monocyte cells, engineered T-cell receptors, engineered B-cells, tumor infiltrating lymphocytes, induced pluripotent stem cells, mesenchymal stem cells, dendritic cells, and hematopoietic stem cells.

[0117] Aspect 28. The modified cells of any one of Aspects 26-27, wherein the target cells are modified using a vector selected from the group consisting of lentivirus, adeno associated virus, virus-like particles, transposon / transposase, transcription activator-like effector nuclease, zinc finger nucleases, mRNA, shRNA, siRNA, SeekRNA, CRISPR / Cas, base editors, prime editors, and programmable addition via site-specific targeting elements.

[0118] Aspect 29. The modified cells of any one of Aspects 26-28, wherein the modified cells are produced in an environment selected from the group consisting of a patient treatment room of an inpatient facility, a patient treatment room of an outpatient facility, a cell processing lab of a hospital facility, a cell processing lab of a non-hospital facility, a cell processing lab of a centralized commercial production facility, a cell processing lab of a decentralized commercial production facility, a sterile manufacturing suite of a hospital facility, a sterile manufacturing suite of a non-hospital facility, a sterile manufacturing suite of an academic facility, a sterile manufacturing suite of a centralized commercial production facility, and a sterile manufacturing suite of a decentralized commercial production facility.

[0119] Aspect 30. The modified cells of any one of Aspects 26-29, wherein said disease, ailment, or medical condition is selected from the group consisting of solid tumors, melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic leukemia, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, small-cell lungcancer, renal transplantation, neuroblastoma, glioblastoma, prostate cancer, hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, and rheumatoid arthritis.

[0120] Aspect 31 . Modified cells for use in the treatment of a disease, ailment, or medical condition of a patient, characterized in that the modified cells are produced using a method of operating a biological fluid processing system for cell affinity selection, wherein the system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container filled with a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a process container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit, the method including pre-processing, processing, and post-processing states; the pre-processing state further comprising: selecting a procedure protocol; procedure setup; installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor at least a first solution container comprising a separation / processing buffer, a second solution container comprising a resuspension buffer and a third solution container comprising a releasing buffer; running a fluid circuit solution prime; attaching to the fluid processor the source container; and running a fluid circuit source prime; the processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid, rinsing of the source container, dilution of the harvested cells, collecting the harvested cells in the processing container, draining buffer solution from the selection column, diluting the material in the selection column via adding antibodies to the selection column, incubating with the antibodies in the selection column, transferring the harvested cells from the processing container into the selection column, transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste, diluting the material in the selection column via transferring biotin into the selection column, incubating, repriming the separation chamber for processing material in the selection column, loading the target fraction from the selection column into the separation chamber, washing andharvesting the cells processed in the separation chamber, rinsing the source container, harvesting selected cells as a final product in at least one final container, and diluting of the harvested cells in the final container; and the post-processing state further comprising: sealing the at least one final container and removing the final product, and removing the disposable fluid circuit.

[0121] Aspect 32. The modified cells of Aspect 31 , wherein the target cells are selected from the group consisting of chimeric antigen receptor T-cells, chimeric antigen receptor natural killer cells, chimeric antigen receptor monocyte cells, engineered T-cell receptors, engineered B-cells, tumor infiltrating lymphocytes, induced pluripotent stem cells, mesenchymal stem cells, dendritic cells, and hematopoietic stem cells.

[0122] Aspect 33. The modified cells of any one of Aspects 31 -32, wherein the target cells are modified using a vector selected from the group consisting of lentivirus, adeno associated virus, virus-like particles, transposon / transposase, transcription activator-like effector nuclease, zinc finger nucleases, mRNA, shRNA, siRNA, SeekRNA, CRISPR / Cas, base editors, prime editors, and programmable addition via site-specific targeting elements.

[0123] Aspect 34. The modified cells of any one of Aspects 31 -33, wherein the modified cells are produced in an environment selected from the group consisting of a patient treatment room of an inpatient facility, a patient treatment room of an outpatient facility, a cell processing lab of a hospital facility, a cell processing lab of a non-hospital facility, a cell processing lab of a centralized commercial production facility, a cell processing lab of a decentralized commercial production facility, a sterile manufacturing suite of a hospital facility, a sterile manufacturing suite of a non-hospital facility, a sterile manufacturing suite of an academic facility, a sterile manufacturing suite of a centralized commercial production facility, and a sterile manufacturing suite of a decentralized commercial production facility.

[0124] Aspect 35. The modified cells of any one of Aspects 31 -34, wherein said disease, ailment, or medical condition is selected from the group consisting of solid tumors, melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic leukemia, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, small-cell lung cancer, renal transplantation, neuroblastoma, glioblastoma, prostate cancer,hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, and rheumatoid arthritis.

Claims

CLAIMS:1 . A biological fluid processing system for cell affinity selection, including a fluid processor having reusable hardware that operates on a disposable fluid circuit, the fluid circuit being connectable to a source container and having a biological fluid, a separation chamber configured to separate the biological fluid from the source container into at least two volumes of material, a processing container for collecting washed and harvested cells from the separation chamber, and a selection column connected to the fluid circuit for further cell affinity selection processing of the cells from the process container.

2. The system of claim 1 , wherein the fluid processor further comprises a controller configured to operate the reusable hardware while the disposable fluid circuit is installed on the reusable hardware and is connected to the source container, the processing container and the selection column.

3. The system of claim 2, wherein the controller further comprises at least a microprocessor and memory.

4. The system of any one of claims 1 -3, wherein the fluid processor further comprises an input device and an output device connected to the controller.

5. The system of any one of claims 1 -4, wherein the reusable hardware of the fluid processor further comprises a drive for a spinning membrane separator in the separation chamber.

6. The system of any one of claims 1 -5, wherein the reusable hardware of the fluid processor further comprises pumps and a cassette interface that engage the fluid circuit.

7. The system of any one of claims 1 -6, wherein the fluid circuit further comprises a single final product container.

8. The system of any one of claims 1 -7, wherein the fluid circuit further comprises a plurality of final product containers.

9. The system of any one of claims 1 -8, wherein the selection column initially contains a matrix or resin material.

10. The system of any one of claims 1 -9, wherein at least three further containers are connected to the fluid circuit, comprising a first solution container comprising aseparation / processing buffer, a second solution container comprising a resuspension buffer and a third solution container comprising a releasing buffer.11 . The system of any one of claims 1 -10, wherein the controller is coupled to a separator drive for a spinning membrane separator of the separation chamber, the at least two syringe pumps and a control cassette interface, the controller configured to selectively operate the separator drive, the syringe pumps and the cassette interface to provide a procedure utilizing the selection column according to a protocol.

12. A method of operating a biological fluid processing system for cell affinity selection, the method comprising: obtaining a biological fluid processing system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container having a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a processing container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit; conducting a pre-processing state further comprising: selecting a procedure protocol, conducting procedure setup, installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor a plurality of containers; running a fluid circuit solution prime including the separation chamber; attaching to the fluid processor the source container; and running a fluid circuit source prime; conducting a processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid; rinsing the source container; diluting the harvested cells; collection of the harvested cells in the processing container; draining solution from the selection column; adding antibodies to the selection column; transferring the harvested cells from the processing container into the selection column; transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste; diluting the material in the selection column; repriming the separation chamber for processing the material in the selection column; target fraction loading from the selection column into the separation chamber; washing and harvesting cells from the selection column; harvesting selected cells as a final product in at least one final container; andconducting a post-processing state further comprising; sealing the at least one final container and removing the final product; and removing the disposable fluid circuit.

13. The method of claim 12, wherein the processing utilizes only negative selection relative to target cells.

14. The method of claim 12 or 13, wherein the processor is configured to further perform positive selection incubation and removal of positive fraction relative to target cells.

15. The method of any one of claims 12-14, wherein the final product is aliquoted into a plurality of smaller containers before removing the fluid circuit.

16. The method of any one of claims 12-15, wherein the processor transfers a negative fraction to waste from the selection column, followed by dilution of the material in the selection column with a releasing buffer.

17. The method of any one of claims 12-16, wherein the releasing buffer is biotin.

18. The method of any one of claims 12-17, wherein the processor further comprises a controller configured to selectively operate the processor to provide a procedure utilizing the separation chamber, processing container and selection column according to a protocol.

19. A method of operating a biological fluid processing system for cell affinity selection, wherein the system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container filled with a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a process container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit, the method comprising: pre-processing, processing, and post-processing states; the pre-processing state further comprising: selecting a procedure protocol; procedure setup; installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor at least a first solution container comprising a separation / processing buffer, a second solution container comprising a resuspension buffer and a third solution container comprising a releasing buffer; running a fluid circuit solution prime; attaching to the fluid processor the source container; and running a fluid circuit source prime;the processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid, rinsing of the source container, dilution of the harvested cells, collecting the harvested cells in the processing container, draining buffer solution from the selection column, diluting the material in the selection column via adding antibodies to the selection column, incubating with the antibodies in the selection column, transferring the harvested cells from the processing container into the selection column, transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste, diluting the material in the selection column via transferring biotin into the selection column, incubating, repriming the separation chamber for processing material in the selection column, loading the target fraction from the selection column into the separation chamber, washing and harvesting the cells processed in the separation chamber, rinsing the source container, harvesting selected cells as a final product in at least one final container, and diluting of the harvested cells in the final container; and the post-processing state further comprising: sealing the at least one final container and removing the final product, and removing the disposable fluid circuit.

20. The method of claim 19, wherein the fluid processor further comprises a controller configured to selectively operate the fluid processor to provide a procedure utilizing the separation chamber, processing container and selection column according to a protocol.21 . Modified cells for use in the treatment of a disease, ailment, or medical condition of a patient, characterized in that the modified cells are produced using a biological fluid processing system, including a fluid processor having reusable hardware that operates on a disposable fluid circuit, the fluid circuit being connectable to a source container and having a biological fluid, a separation chamber configured to separate the biological fluid from the source container into at least two volumes of material, a processing container for collecting washed and harvested cells from the separation chamber, and a selection column connected to the fluid circuit for further cell affinity selection processing of the cells from the process container.

22. The modified cells of claim 21 , wherein the target cells are selected from the group consisting of chimeric antigen receptor T-cells, chimeric antigen receptor natural killer cells, chimeric antigen receptor monocyte cells, engineered T-cell receptors, engineered B-cells, tumor infiltrating lymphocytes, induced pluripotent stem cells, mesenchymal stem cells, dendritic cells, and hematopoietic stem cells.

23. The modified cells of any one of claims 21 -22, wherein the target cells are modified using a vector selected from the group consisting of lentivirus, adeno associated virus, virus-like particles, transposon / transposase, transcription activator-like effector nuclease, zinc finger nucleases, mRNA, shRNA, siRNA, SeekRNA, CRISPR / Cas, base editors, prime editors, and programmable addition via site-specific targeting elements.

24. The modified cells of any one of claims 21 -23, wherein the modified cells are produced in an environment selected from the group consisting of a patient treatment room of an inpatient facility, a patient treatment room of an outpatient facility, a cell processing lab of a hospital facility, a cell processing lab of a non-hospital facility, a cell processing lab of a centralized commercial production facility, a cell processing lab of a decentralized commercial production facility, a sterile manufacturing suite of a hospital facility, a sterile manufacturing suite of a non-hospital facility, a sterile manufacturing suite of an academic facility, a sterile manufacturing suite of a centralized commercial production facility, and a sterile manufacturing suite of a decentralized commercial production facility.

25. The modified cells of any one of claims 21 -24, wherein said disease, ailment, or medical condition is selected from the group consisting of solid tumors, melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic leukemia, nonHodgkin lymphoma, diffuse large B-cell lymphoma, small-cell lung cancer, renal transplantation, neuroblastoma, glioblastoma, prostate cancer, hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, and rheumatoid arthritis.

26. Modified cells for use in the treatment of a disease, ailment, or medical condition of a patient, characterized in that the modified cells are produced using a method comprising: obtaining a biological fluid processing system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container having a biological fluid and configured to separate the biological fluid from the source container into at least two volumes of material, a processing container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit; conducting a pre-processing state further comprising: selecting a procedure protocol, conducting procedure setup, installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor a plurality of containers; running a fluid circuit solution prime including the separation chamber; attaching to the fluid processor the source container; and running a fluid circuit source prime; conducting a processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid; rinsing the source container; diluting the harvested cells; collection of the harvested cells in the processing container; draining solution from the selection column; adding antibodies to the selection column; transferring the harvested cells from the processing container into the selection column; transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste; diluting the material in the selection column; repriming the separation chamber for processing the material in the selection column; target fraction loading from the selection column into the separation chamber; washing and harvesting cells from the selection column; harvesting selected cells as a final product in at least one final container; and conducting a post-processing state further comprising; sealing the at least one final container and removing the final product; and removing the disposable fluid circuit.

27. The modified cells of claim 26, wherein the target cells are selected from the group consisting of chimeric antigen receptor T-cells, chimeric antigen receptor natural killer cells, chimeric antigen receptor monocyte cells, engineered T-cell receptors,engineered B-cells, tumor infiltrating lymphocytes, induced pluripotent stem cells, mesenchymal stem cells, dendritic cells, and hematopoietic stem cells.

28. The modified cells of any one of claims 26-27, wherein the target cells are modified using a vector selected from the group consisting of lentivirus, adeno associated virus, virus-like particles, transposon / transposase, transcription activator-like effector nuclease, zinc finger nucleases, mRNA, shRNA, siRNA, SeekRNA, CRISPR / Cas, base editors, prime editors, and programmable addition via site-specific targeting elements.

29. The modified cells of any one of claims 26-28, wherein the modified cells are produced in an environment selected from the group consisting of a patient treatment room of an inpatient facility, a patient treatment room of an outpatient facility, a cell processing lab of a hospital facility, a cell processing lab of a non-hospital facility, a cell processing lab of a centralized commercial production facility, a cell processing lab of a decentralized commercial production facility, a sterile manufacturing suite of a hospital facility, a sterile manufacturing suite of a non-hospital facility, a sterile manufacturing suite of an academic facility, a sterile manufacturing suite of a centralized commercial production facility, and a sterile manufacturing suite of a decentralized commercial production facility.

30. The modified cells of any one of claims 26-29, wherein said disease, ailment, or medical condition is selected from the group consisting of solid tumors, melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic leukemia, nonHodgkin lymphoma, diffuse large B-cell lymphoma, small-cell lung cancer, renal transplantation, neuroblastoma, glioblastoma, prostate cancer, hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, and rheumatoid arthritis.31 . Modified cells for use in the treatment of a disease, ailment, or medical condition of a patient, characterized in that the modified cells are produced using a method of operating a biological fluid processing system for cell affinity selection, wherein the system comprises a fluid processor that operates on a disposable fluid circuit that is connectable to a source container filled with a biological fluid and configured to separatethe biological fluid from the source container into at least two volumes of material, a process container connected to the fluid processor along a fluid pathway, and a selection column connected to the fluid circuit, the method comprising: pre-processing, processing, and post-processing states; the pre-processing state further comprising: selecting a procedure protocol; procedure setup; installing the disposable fluid circuit on the fluid processor; attaching to the fluid processor at least a first solution container comprising a separation / processing buffer, a second solution container comprising a resuspension buffer and a third solution container comprising a releasing buffer; running a fluid circuit solution prime; attaching to the fluid processor the source container; and running a fluid circuit source prime; the processing state further comprising: processing the biological fluid from the source container in a separation chamber via the disposable fluid circuit, including one or more cycles of washing, harvesting and dilution of the biological fluid, rinsing of the source container, dilution of the harvested cells, collecting the harvested cells in the processing container, draining buffer solution from the selection column, diluting the material in the selection column via adding antibodies to the selection column, incubating with the antibodies in the selection column, transferring the harvested cells from the processing container into the selection column, transferring a non-target fraction comprising unbound cells that remain in suspension in the selection column to waste, diluting the material in the selection column via transferring biotin into the selection column, incubating, repriming the separation chamber for processing material in the selection column, loading the target fraction from the selection column into the separation chamber, washing and harvesting the cells processed in the separation chamber, rinsing the source container, harvesting selected cells as a final product in at least one final container, and diluting of the harvested cells in the final container; and the post-processing state further comprising: sealing the at least one final container and removing the final product, and removing the disposable fluid circuit.

32. The modified cells of claim 31 , wherein the target cells are selected from the group consisting of chimeric antigen receptor T-cells, chimeric antigen receptor natural killer cells, chimeric antigen receptor monocyte cells, engineered T-cell receptors,engineered B-cells, tumor infiltrating lymphocytes, induced pluripotent stem cells, mesenchymal stem cells, dendritic cells, and hematopoietic stem cells.

33. The modified cells of any one of claims 31 -32, wherein the target cells are modified using a vector selected from the group consisting of lentivirus, adeno associated virus, virus-like particles, transposon / transposase, transcription activator-like effector nuclease, zinc finger nucleases, mRNA, shRNA, siRNA, SeekRNA, CRISPR / Cas, base editors, prime editors, and programmable addition via site-specific targeting elements.

34. The modified cells of any one of claims 31 -33, wherein the modified cells are produced in an environment selected from the group consisting of a patient treatment room of an inpatient facility, a patient treatment room of an outpatient facility, a cell processing lab of a hospital facility, a cell processing lab of a non-hospital facility, a cell processing lab of a centralized commercial production facility, a cell processing lab of a decentralized commercial production facility, a sterile manufacturing suite of a hospital facility, a sterile manufacturing suite of a non-hospital facility, a sterile manufacturing suite of an academic facility, a sterile manufacturing suite of a centralized commercial production facility, and a sterile manufacturing suite of a decentralized commercial production facility.

35. The modified cells of any one of claims 31 -34, wherein said disease, ailment, or medical condition is selected from the group consisting of solid tumors, melanoma, multiple myeloma, acute lymphoblastic leukemia, acute myelocytic leukemia, nonHodgkin lymphoma, diffuse large B-cell lymphoma, small-cell lung cancer, renal transplantation, neuroblastoma, glioblastoma, prostate cancer, hemophilia, sickle cell disease, lupus erythematosus, lupus nephritis, myasthenia gravis, autoimmune diseases, soft tissue sarcoma, osteosarcoma, hepatocellular carcinoma, graft versus host disease, and rheumatoid arthritis.

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