Blood Processing Systems

Abstract

Blood Processing Systems The present invention relates to a system for collecting, separating, concentrating, modifying and reinfusing blood components from whole blood. The system includes a blood treatment device and a fluid flow circuit.

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 484,636, filed February 13, 2023, the contents of which are incorporated herein by reference. This disclosure relates to blood processing. More particularly, this disclosure relates to systems and methods for collecting blood components and processing or modifying cells prior to reinfusion. [Background technology]

[0002] Various blood processing systems allow for the separation of blood into two or more components, which may be useful for donation purposes or for treating individuals with harmful or potentially harmful conditions or diseases.

[0003] When such systems are used for blood component donations, whole blood is typically drawn from a donor, certain blood components or constituents are removed and collected, and the remaining blood components are returned to the donor.

[0004] Such systems can also be used to provide blood components for cell therapy to patients or individuals. These therapies typically involve separating specific cellular or other blood components from whole blood, modifying, enriching, and / or expanding the collected components, and then returning them to the patient as part of the treatment. For example, a treatment known as chimeric antigen receptor (CAR) T-cell therapy involves modifying a patient's T cells by attaching artificial receptors that bind to antigens on cancer cells. The modified T cells can then be returned to the patient and used to target and destroy specific cancer cells.

[0005] Modified therapeutic cells are typically produced in manufacturing facilities separate from the blood collection site, a process that can be time-consuming and burdensome, resulting in a substantial time gap between when the cells are collected and when the modified or processed cells are re-infused.

[0006] Although treatments such as CAR-T therapy have shown high clinical efficacy against hematologic malignancies, many patients are ineligible for treatment due to the long intervenous time required for administration of these autologous gene-modified therapies. Therefore, there is a need for rapid production and retransfusion of therapeutic cells, including but not limited to gene-modified autologous cells.

[0007] It is therefore desirable to provide an integrated system capable of collecting, concentrating, modifying, preparing for reinfusion, and / or reinfusing cells. Summary of the Invention

[0008] The present subject matter includes multiple aspects that may be implemented individually or in combination in the devices and systems described and claimed below. These aspects may be implemented alone or in combination with other aspects described herein, and the description of these aspects together is not intended to preclude the aspects from being used individually or from being claimed individually or in different combinations, as recited in the appended claims.

[0009] In one embodiment, a fluid flow circuit used in a blood processing system includes a separation module, at least one pump, at least one fluid reservoir for holding fluid during blood processing, a valve system, at least one fluid supply container, at least one cell modification module, a blood source access device, and a plurality of conduits fluidly connecting the components of the fluid flow circuit. [Brief explanation of the drawings]

[0010] FIG. 1 is a schematic diagram of an exemplary fluid flow circuit.

[0011] FIG. 2 is a schematic diagram of a blood processing device.

[0012] FIG. 3A is a perspective view of an exemplary fluid flow circuit.

[0013] FIG. 3B is a perspective view of the blood processing device in an open state with the fluid flow circuit inserted.

[0014] FIG. 3C is a perspective view of the blood processing system.

[0015] FIG. 4 is a front perspective view of an exemplary fluid flow circuit.

[0016] FIG. 5 is a rear perspective view of an exemplary fluid flow circuit.

[0017] FIG. 6 is a schematic diagram of an exemplary first portion of a disposable fluid flow circuit.

[0018] FIG. 7 is a schematic diagram of another exemplary first portion of a disposable fluid flow circuit.

[0019] FIG. 8 is a schematic diagram of an exemplary second portion of a disposable fluid flow circuit.

[0020] FIG. 9 is a schematic diagram of the priming process in the disposable fluid flow circuit of FIG.

[0021] FIG. 10 is a schematic diagram of the first separation step in the disposable fluid flow circuit of FIG.

[0022] FIG. 11 is a schematic diagram of the second separation step in the disposable fluid flow circuit of FIG.

[0023] FIG. 12 is a schematic diagram of the cell concentration process in the disposable fluid flow circuit of FIG.

[0024] FIG. 13 is a schematic diagram of the first cell preparation step in the disposable fluid flow circuit of FIG.

[0025] FIG. 14 is a schematic diagram of a second cell preparation step in the disposable fluid flow circuit of FIG.

[0026] FIG. 15 is a schematic diagram of the gene transfer process in the disposable fluid flow circuit of FIG.

[0027] FIG. 16 is a schematic diagram of the cell selection process in the disposable fluid flow circuit of FIG. DETAILED DESCRIPTION OF THE INVENTION

[0028] The embodiments disclosed herein are intended to be illustrative of the technology, and it is understood that the technology may be embodied in other forms or combinations not specifically shown. Therefore, the specific designs and features disclosed herein should not be construed as limiting the technology defined by the appended claims.

[0029] The present disclosure will be more fully understood when taken in conjunction with the following description and the accompanying drawings. Some of the figures may be simplified by omitting selected elements to more clearly show other elements. The omission of elements in some of the figures does not indicate the presence or absence of a particular element in any illustrative embodiment unless expressly stated in the corresponding description. The figures are not necessarily drawn to scale.

[0030] The present disclosure includes exemplary embodiments of fluid flow circuits and blood processing devices that can be combined to form an automated blood processing system for collecting, separating, concentrating, and modifying blood cells for reinfusion into a patient.

[0031] "Blood" includes, but is not limited to, blood and blood components, and "cells" or "biological cells" includes, but is not limited to, blood cells, such as red blood cells, white blood cells, and T-cells. "Automated" means that the device is programmable to perform each step in the method for processing biological fluids without substantial operator involvement. Of course, it is understood that the automated system of the present disclosure still involves some operator interaction, such as loading the disposable fluid flow circuit and inputting processing parameters. Additional manual steps may be required. However, via the disposable circuit described below, the reusable device is capable of processing blood without substantial operator involvement.

[0032] As shown in Figures 1 and 2, the blood processing system includes two major components: a durable, reusable blood processing device 20 (Figure 2) and a disposable fluid flow circuit (Figure 1), collectively referred to herein as element 10. The blood processing device includes components that control and monitor fluid flow within the disposable fluid flow circuit 10, and a controller 16 (Figure 3C) that controls and / or directs the operation of the other components of the blood processing device 20 to perform the blood processing procedure selected by the operator, as described below.

[0033] The blood processing systems and methods according to the present disclosure are described as using blood processing devices or systems and various cell modification modules. However, it should be understood that the principles described herein are not limited to any particular configuration of device or to any particular sequence of steps or steps. Rather, the blood processing systems and methods described herein can be used with different configurations of blood processing devices and fluid flow circuits to perform blood processing procedures in different ways.

[0034] As shown in Figures 2, 3B, and 3C, blood processing device 20 can be modular and can include multiple components designed to process blood components in conjunction with various fluid flow circuits 10. The device can include movable parts or components, such as wheels, for transport to and from a patient's bed or chair. The blood processing device can include valves or valve components, pneumatic control systems, pumps, detectors, sensors, controls, user interfaces, and other components that can function to assist in the movement of fluid within fluid flow circuit 10.

[0035] Blood processing device 20 can include valves or motors associated with the valves on fluid flow circuit 10. The valves can be configured to interact with the conduits of fluid flow circuit 10 attached to device 20. By way of example, the valves can be solenoid pinch valves, motor-driven rotary pinch valves, linear actuators, stopcocks, or any other type of automated clamping or valving device known in the art. In an exemplary embodiment, blood processing device 20 includes valve motors that are compatible with the valve components on fluid flow circuit 10.

[0036] The blood processing device 20 can include an air control system for supplying air to, for example, a pneumatic syringe pump assembly, described below. The air control system can include a vacuum and / or pressure source, such as a diaphragm pump, that can deliver filtered air to and from the pump. Examples and details of assemblies that can be used in the blood processing device 20 are described in U.S. Patent Publication Nos. 10,926,895; 11,191,880; 10,781,001; and U.S. Patent Application No. 17 / 026,156.

[0037] The blood processing device 20 may include multiple pumps (two possible pumps 30, 32 are shown in FIG. 6) for circulating fluid through the fluid flow circuit 10. The pumps may be of different or similar configurations and / or may have different or similar functions. In one embodiment, the pumps may be configured as peristaltic pumps of the general configuration described in U.S. Pat. No. 5,868,696. Each pump engages a different line and may be selectively activated, under command of the controller 16, to circulate fluid through portions of the fluid flow circuit 10.

[0038] The illustrated blood processing device 20 can include an air detector (e.g., an ultrasonic air bubble detector) associated with the tubing of the fluid flow circuit 10 that carries fluid toward the recipient. The air detector is labeled "A" in the fluid flow circuit schematic of FIG. 8. Preventing air from reaching the recipient can be advantageous, whether the recipient is human (e.g., the same human as the source of the blood) or non-human (e.g., a storage bag or container). Accordingly, the air detector can send a signal to the controller 16 indicating the presence or absence of air in the tubing. If the signal indicates the presence of air in the tubing, the controller 16 can initiate an alarm or error state to alert an operator of the condition and / or take corrective action (e.g., reversing fluid flow in the tubing or diverting the flow to an air vent) to prevent air from reaching the recipient. The air detector can alternatively or additionally be used as part of the fluid flow control of the fluid flow circuit 10.

[0039] The illustrated blood processing device 20 may include one or more sensors or sensing elements for detecting the state or characteristics of blood components. For example, in one embodiment, a cell concentration sensor 78 (shown on line L1 in FIGS. 8-16) may be incorporated to detect absolute or relative changes in cell concentration. Optical devices using techniques such as light transmission, scattering, or spectroscopy may be used. Devices using electrical techniques such as capacitance may also be useful. Furthermore, devices using acoustic techniques may also be capable of providing relative concentration measurements.

[0040] As shown in FIG. 8 , the plunger position of any of the syringe pumps 54, 56, 58, 60 included in fluid flow circuit 10 may be tracked by sensing elements. Additionally, pressure sensors may be incorporated into the system to monitor pressure at various points in fluid flow circuit 10. For example, if the blood source is a human donor, one or more pressure sensors, such as donor pressure sensor 34 ( FIG. 6 ), may be configured to monitor the pressure in the donor's vein during withdrawal and return. Controller 16 may receive signals from the pressure sensors indicative of the pressure within fluid flow circuit 10 and, if the signals indicate a low-pressure or high-pressure condition, may initiate an alarm or error condition to alert an operator of the condition and / or to restore the pressure to an acceptable range without operator intervention.

[0041] As mentioned above, blood processing device 20 includes a controller, such as controller 16 shown in Figure 3C. The controller is shown on top of blood processing device 20, but may also be incorporated elsewhere in blood processing device 20. According to embodiments described herein, the controller may include a programmable microprocessor, which may be programmed to operate blood processing device 20 and system 21 according to a predetermined process.

[0042] According to other embodiments, the controller may include one or more electrical circuits designed to perform the operations described herein. Additionally, the controller may include one or more memory devices. Instructions for programming the microprocessor may be stored in memory devices associated with the microprocessor. The memory devices may include one or more tangible, non-transitory computer-readable storage media having computer-executable instructions stored thereon that, when executed by the microprocessor, enable the microprocessor to perform one or more of the operations described below.

[0043] The controller is connected to one or more of the components of the blood processing device 20 and the fluid flow circuit 10 (FIG. 8) and can receive information (e.g., in the form of signals) from these components and send commands (e.g., in the form of signals) to these components to control their operation. The controller is connected to sensors, valves, and pumps and can send commands to control the operation of these devices. The controller can also receive information from and send commands to specific components, such as those mentioned above. The controller may be connected to these components by being directly electrically connected to them, or may be connected to these components by being directly connected to other intermediate devices that are directly connected to these components.

[0044] The control device is configured and / or programmed to perform at least one blood processing procedure (such as those shown in Figures 9-16), but more preferably is configured and / or programmed to perform multiple blood processing procedures, which may include separation and cell modification steps.

[0045] More specifically, in performing any blood processing procedure, the controller is configured and / or programmed to control the flow and volume of fluids from one component to another. This may include directing valves to open or close at specific times during the procedure, or initiating the transfer of fluids from one container to another. Thus, even if a particular component of the blood processing system is described herein as performing a particular function, it should be understood that the component is controlled by the controller to initiate and / or perform that function.

[0046] A user interface screen 14 (e.g., a touchscreen) can be associated with blood processing device 20 (see FIG. 3C). User interface screen 14 allows an operator to interact with a system controller (e.g., a microprocessor) of device 20, providing commands to the controller (e.g., to perform a particular procedure) and information to be used during the procedure (e.g., a white blood cell (WBC) pre-count of the blood source). User interface screen 14 can provide instructions (e.g., instructions to connect or disconnect the blood source to flow circuit 10) and information (e.g., information alerting the operator to an occlusion in a fluid flow conduit of flow circuit 10) to the operator and can function as a display to show the progress of a procedure.

[0047] The blood processing device 20 may include computer equipment that allows the blood processing device 20, including the controller 16, to communicate with other blood processing devices on a local network, or with other blood processing devices or other computer equipment (e.g., servers) via a local network, a wide area network, or the Internet, either wired (wires, cables, etc.) or wirelessly. According to such an embodiment, the device may include an internal transceiver.

[0048] 3A-3C illustrate the steps of inserting and loading fluid flow circuit 10 into blood processing device 20 to construct blood processing system 21. Fluid flow circuit 10 (FIG. 3A) is inserted into the open cabinet of the blood processing device (FIG. 3B), components are connected as needed, and then the cabinet is closed in a third step (FIG. 3C). Blood processing system 21, including blood processing device 20 and fluid flow circuit 10, is a single modular system capable of performing an entire blood processing procedure without traveling to external components or devices.

[0049] The fluid flow circuit or flow set 10 (shown in detail in Figures 4-8) is intended to be sterile, single-use, and disposable. Figures 4 and 5 include perspective views of the fluid flow circuit 10, and Figure 8 shows a schematic diagram of the fluid flow circuit. Figures 9-16 illustrate steps in an exemplary procedure. The fluid flow circuit is modular, allowing for different customized configurations that can be loaded into the blood processing device 20. Before initiating a given blood processing and modification procedure, the operator loads the fluid flow circuit 10 into the blood processing device 20. The controller 16 executes the procedure based on a pre-set protocol, taking into account other inputs from the operator. After the procedure is complete, the operator removes the fluid flow circuit 10 from the blood processing device 20. If any portions of the fluid flow circuit 10 retain components (e.g., doses), they are removed from the device 20 and retained for storage, transfusion, or further processing. The remaining fluid flow circuit 10 is removed from the blood processing device 20 and discarded.

[0050] The various fluid flow circuits used in conjunction with blood processing devices may have slightly different components depending on the blood processing procedure being performed and the type of cells being processed by the system. Thus, different fluid flow circuits may be used in connection with specific blood processing procedures. Generally, fluid flow circuit 10 may include a pump, a reservoir, valve components, fluid input and output vessels, a separator, a concentrator, and at least one cell modification module (as shown in FIG. 8), or a combination thereof.

[0051] The fluid flow circuit can include two distinct portions: a first, near-user portion (22 or 23 shown in Figures 6 and 7) and a second, processing portion 25 (shown in Figure 8). The near-user portion can be selected depending on whether the processing is active (re-infusion into a donor) or passive (collecting a dose for later infusion). Figure 6 shows possible components and a schematic diagram of active processing. Figure 7 shows possible components and a schematic diagram of passive processing.

[0052] Both near-user sections include at least one blood source access device 26 (e.g., a blood collection needle) used to withdraw blood from and deliver fluids to the blood source. Optionally, two blood source access devices (e.g., dual needles) may be used, one used to draw blood from the source into the fluid flow circuit 10 and the other used to return fluid to the source. In another embodiment, a blood source (e.g., a pre-collection bag) may be connected to the system. Both near-user sections also include a donor isolation clamp 28. A main line L1 is present in both sections and connects the fluid processing section 25 (FIG. 8) of the fluid flow circuit 10 to the near-user section.

[0053] In the active near-user portion 22 of the fluid flow circuit 10 shown in Figure 6, two independent flow paths are connected to the blood source access device 26, with a first line L3 connected to the anticoagulant container 24 and a second line L2 connected to the reservoir 40. The blood processing device 20 can include an associated anticoagulant pump 30 and a blood withdrawal / return pump 32 for moving fluids to and from the patient.

[0054] 7 is only connected to the reservoir 40 and optionally the level sensing element 38, and therefore includes only line L1. Prior to treatment, an anticoagulant may be added to the reservoir 40.

[0055] The passive or active near-user portion may include multiple pumps (two possible pumps 30, 32 are shown in FIG. 6) for circulating fluid through the fluid flow circuit 10. The pumps may be different or similar in configuration and / or different or similar in function. In one embodiment, the pumps are configured as peristaltic pumps generally configured as described in U.S. Pat. No. 5,868,696. Each pump engages a different line and can be selectively activated, upon command of the controller 16, to circulate fluid through portions of the fluid flow circuit 10.

[0056] Turning now to the fluid processing portion 25, FIG. 8 illustrates an example of the fluid flow circuit 10. The fluid flow circuit 10 can include multiple fluid input and output containers. Each container can be integrally formed with the fluid flow circuit 10 or can be connected to the fluid flow circuit (e.g., by puncturing a septum in the fluid flow circuit tubing, via a Luer connector, or by aseptically joining using a sterile welding system). These can be connected before the fluid flow circuit is connected to the blood processing device to form the blood processing system 21. The containers can be constructed of any desired medical-grade material, such as medical-grade plastic. The fluid input containers included in the processing portion 25 of the fluid flow circuit can include at least one buffer container 46 and at least one solution or liquid container 64. The solution or liquid container can be configured to hold a liquid chemical composition for mixing with blood or blood cell components. An optional first buffer container 46 and a second buffer container 48, as well as four optional solution or liquid containers 64, 66, 68, and 70, are shown in FIG. 8. However, the number of buffer reservoirs and the number and / or presence of solution or liquid reservoirs may vary depending on the cell modification treatment used.

[0057] Output containers may also be integrally formed with or connected to fluid flow circuit 10. These containers are intended to contain various cell fractions, spent buffers, prepared suspensions, or samples thereof. These containers may include waste container 52 or final dose container 50, which are also shown in FIG. 8. In cases where processing occurs in which the dose is not immediately returned to the patient (and removed from the system) after modification, dose container 50 may be a removable container that is transported after processing.

[0058] The fluid flow circuit may include a valve or valve arrangement (V1-V21 shown in FIGS. 8-16). The valves may be stopcock valves. These valves cooperate or interface with motors that are part of the blood processing device's hardware. The valves may be used to direct fluid flow between different components of the fluid flow circuit.

[0059] The fluid flow circuit can include pumps 54, 56, 58, and 60. While four pumps are shown in FIG. 8, the fluid flow circuit can use more or fewer pumps. The pumps are preferably configured as pneumatic syringe pump assemblies that interface with a pneumatic control system in the blood processing device 20 hardware. Positive or negative pressure can be applied to displace the syringe plungers. Positive pressure means fluid flows out of the pump, and negative pressure means fluid flows into the pump. Optionally, the syringe caps may have sterile filters embedded in them. In one embodiment, the pumps can be generally configured as described in U.S. Patent Application No. 2021 / 0121827, which is incorporated herein by reference in its entirety. The plunger position can be tracked by sensing elements on the hardware. The pumps can be operated in a pressure-targeted mode or a flow-targeted mode, depending on the control scheme required for the process.

[0060] The fluid flow circuit can include multiple reservoirs 40, 42, 44. The reservoirs are used as passive containers to hold fluids before, during, and after processing steps. The reservoirs may be vented with sterile filters so that fluid flow into or out of the reservoirs does not cause pressurization of the container. While FIG. 8 shows a fluid flow circuit with three reservoirs, fluid flow circuit 10 may include more or fewer reservoirs.

[0061] The fluid flow circuit 10 can include a separation module 62 (see FIG. 8). The microfluidic separation module 62 can be used to continuously separate particles or cells. The microfluidic separation module can include multiple channels for separating cells based on their characteristics (e.g., diameter). For example, a critical diameter of approximately 7 μm can separate nucleated white blood cells from red blood cells and platelets. The separation module 62 shown in FIG. 8 has two outputs 62c and 62d. The first output 62d is for cells larger than a predetermined critical diameter, and the second output 62c is for cells smaller than a predetermined critical diameter. These separation modules can also be used to transfer target cell populations to a new buffer, essentially "washing" the cell suspension. In another embodiment, the separation module can include a spinning membrane separator or centrifuge chamber used in other blood processing devices. These are described in detail in U.S. Patent Nos. 4,526,515 (DeVries), 5,194,145 (Schoendorfer), 6,312,607 (Brown et al.), 6,524,231 (Westberg et al.), 4,094,461 (Kellogg et al.), 7,052,606 (Gibbs et al.), 4,300,717 (Latham), and 8,075,468, as well as U.S. Patent Application Publication No. 2009 / 0215602 (Min et al.), all of which are incorporated herein by reference. When a spinning membrane separator or centrifuge is used, the blood processing device can include corresponding hardware components.

[0062] The fluid flow circuit 10 can include a concentration module or microfluidic concentration module 72 for continuous particle or cell concentration, thereby continuously producing a concentrated output stream (outlet 72c) and a supernatant output stream (outlet 72b) from a single input stream (inlet 72a) (see FIG. 8). The concentration module 72 can be operated at a fixed concentration factor, such as 10x, per pass. A desired cell concentration can be achieved by performing multiple fixed concentration and dilution steps. As an example, white blood cells can be concentrated 25x. The concentration module can also achieve variable concentrations by using multiple passes and including dilutions between passes. For example, a 15x concentration can be achieved in this manner.

[0063] Fluid flow circuit 10 may be further configured to interface with or include at least one cell modification module. By way of example, these modules may perform cell therapy and / or gene therapy. Thus, fluid flow circuit 10 may be configured to interface with one or more of a gene transfer module and a cell selection module.

[0064] A gene transfer module 74 may be included as part of the fluid flow circuit 10, as shown in FIG. 8. The module may be used to deliver intracellular payloads and cell suspensions. The gene transfer module may include an inlet 74a and an outlet 74b. Examples of modules include electroporators, mechanoporators, sonoporators, sorborators, and other flow-through transfection techniques and / or means for introducing therapeutic payloads.

[0065] A cell selection module 76 (e.g., an affinity-based cell selection module) may be included as part of the fluid flow circuit 10, as shown in FIG. 8. The cell selection module 76 may be used to separate a target cell population from a bulk cell suspension based on phenotype. The cell selection module 76 may include an inlet 76a, which may also function as an outlet. Techniques such as magnetic beads, affinity chromatography, or filtration may be performed using positive or negative selection methods.

[0066] Alternatively, fluid flow circuit 10 may be configured to utilize an existing portion of fluid flow circuit 10 as a cell modification module, such as for cell preparation. In such cases, solutions or liquid chemical additives may be added to the blood cells in reservoirs (e.g., 42, 44) or pumps (e.g., 60) integrated into the fluid flow circuit, or both. The blood cells may be prepared, mixed, or incubated within the reservoirs, or may be transferred between components of fluid flow circuit 10.

[0067] As previously mentioned, the components of fluid flow circuit 10 may be connected by flexible tubing or other suitable fluid communication conduits. Fluid flow circuit 10 includes lines L1-L32 (shown in detail in FIG. 8), which may include more or fewer lines depending on the desired configuration of fluid flow circuit 10 and the modules connected to it.

[0068] The fluid flow circuit may incorporate various additional components, for example, a return line filter may be associated with the line to the fluid receptor, and filters may be placed upstream of one or more fluid containers and used to remove substances (e.g., white blood cells) from the separated components (e.g., red blood cells or platelets).

[0069] Prior to initiating a blood processing procedure, any fluid containers not integrally formed with fluid flow circuit 10 may be connected to fluid flow circuit 10 (e.g., by puncturing the septum of the tubing of fluid flow circuit 12 or via Luer connectors). Fluid flow circuit 10 is then attached to blood separation device 20. Additionally, any inlet containers may be filled with an appropriate fluid, such as a buffer or solution. Controller 16 may perform an integrity check of fluid flow circuit 10 to ensure that each component is properly connected and functioning.

[0070] To initiate a blood processing procedure, the operator may select the desired procedure from among the various procedures that the device 10 can perform (e.g., using the user interface screen 14). The operator may input various information required by the system controller, which may enable the controller to more appropriately execute the procedure. The controller may be provided with the desired cell modification process, the solution to be used, the type of blood cells to be modified, the total blood volume required for processing or the total blood volume of the blood source, a pre-white blood cell count or an initial white blood cell concentration of the blood source, and a post-white blood cell count or a target platelet concentration of the blood source to be achieved at the end of the procedure. The total volume of blood to be processed may also be provided to the system controller. Additionally, various measurements, such as the patient's height and weight, may also be added.

[0071] After the system controller has received all necessary inputs and performed the necessary preliminary calculations and status checks (e.g., verifying that the fluid flow circuit 10 is properly installed and that each component of the system 21 is functioning properly), a blood source is connected to the fluid flow circuit 10 (e.g., by drawing blood from a donor or by connecting a whole blood container), and the blood processing procedure can begin.

[0072] Blood is introduced into the system through active or passive portions of fluid flow circuit 10 located near the operator. Blood flows from a donor into main line L1 and reservoir 40. An initial step, referred to as the "blood prime" step (see FIG. 9), is performed in which selected components of fluid flow circuit 10 are primed with blood 41 from a blood source, particularly as stored in reservoir 40. It is also within the scope of this disclosure for fluid flow circuit 10 to be primed with a different priming fluid, such as saline.

[0073] During the blood prime phase, whole blood is drawn from the reservoir into fluid flow circuit 10 via line L2. The blood travels through line L1 to the pro-patient portion of fluid flow circuit 10. Although not shown in FIG. 9, the priming process may also be performed on other components of fluid flow circuit 10, such as portions of processor 25.

[0074] Next, the separation stage can begin. In the first step of this stage, as shown in FIG. 10, blood 41 fills pump 54, and buffer 47 fills pumps 56 and 58. Blood or blood components 41 from reservoir 40 are drawn into pump 54 through lines L2, L1, valve V1, and line L4. Buffer 47 from container 46 is drawn into pumps 56 and 58. Specifically, buffer is sent from container 46 through line L9 and valve V5 to line L10 (to pump 56 via valve V4 and line L30) or line L11 (to pump 58 via valve V6 and line L31). Buffer 47 may also optionally be sent to pump 54. In the second step of the separation stage, as shown in FIG. 11, pumps 56 and 58 are pressurized to operating pressure and flow into separation module 62. Pumps 56 and 58 may be operated sequentially to provide continuous flow. Buffer 47 is pumped from pump 56 or 58 through line L12, valve V3, and line L6 to inlet 62b of separation module 62. Pump 54 also pumps blood components 55 through line L4, valve V1, valve V2, and line L5 to inlet 62a of separation module 62. Cells are separated based on size. Larger cells / particles 43, such as white blood cells, are directed from outlet 62d through line L8, valves V8, and V7 to line L13 and into reservoir 42. Smaller cells / particles 53, such as red blood cells and platelets, are directed from outlet 62c through line L7, line L1, valve V21, and line 29 to waste container 52. The separation module can operate differently depending on the desired separation and the specific cells to be modified.

[0075] Next, the cell concentration process shown in FIG. 12 can be performed. Large cellular components 43 are drawn from reservoir 42 into pump 60 via line L13, valves V7, V8, V9, and line L14. Pump 60 is then pressurized to its operating pressure, and the large cellular components are pumped through line L14 and valve V9 to line L1, and then through valve V15 and line L22 to inlet 72a of concentration module 72. The concentration module produces concentrated cells 45 and supernatant 59. Supernatant 59 is pumped from outlet 72b through line L20, line L7, valve V21, and line L29 to waste container 52. Concentrated cells 45 are pumped from outlet 72c through line L23, valve V18, line L1, and valve V19 to line L27 and then to reservoir 44. The concentrated cells can then be diluted and, optionally, concentrated again. That is, the cells can be processed by being drawn from the reservoir 44 back into the pump 60 and repeatedly passing through the concentration module 72 until the target concentration is reached. The cell concentration can be detected during concentration or during transfer.

[0076] A cell preparation step or process may also be performed. This process may be performed on concentrated cells 45 (shown in FIGS. 13 and 14) or on separated cellular components (e.g., large cellular components 43). The concentrated cells 45 or large cellular components 43 may be treated with at least one solution, buffer, or combination thereof in reservoirs 42, 44. As an example, in the first step shown in FIG. 13, a specific volume of solution 65 is drawn into pump 60. Solution 65 from reservoir 64 travels through line L15, valves 10, 11, 12, 13, and 14 to lines L21 and L1, and then to valve 9 and line L14. As a second step in the cell preparation process of the exemplary method, as shown in FIG. 14, solution 65 is pumped toward reservoir 44, which may contain concentrated cells 45. Specifically, the solution passes through line L14 and valve V9 to line L1, and then through valves V15, V16, V17, V18, and V19 to line L27. The cells may be mixed with the solution and incubated as a suspension 67. During the holding or incubation step, the suspension may be shuttled between reservoir 44 and pump 60 to prevent settling. As an alternative second step in the cell preparation step (if added to large cellular components 43), solution 65 may be pumped toward reservoir 42, where large cellular components 43 are held after the separation step (see FIG. 11). Specifically, the solution passes through line L14 and valve V9 to line L1, and then through valves V8 and V7 to line L13. The cells may be mixed with the solution and incubated in reservoir 42. During incubation, the suspension may be shuttled between reservoir 42 and pump 60 to prevent settling.

[0077] As shown in FIG. 15, a gene transfer step can also be performed. Cells 69 prepared with a genetic modification solution are pumped through pump 60 to gene transfer module 74, where a payload is introduced into the cells. Similar to the processes shown in FIGS. 13 and 14, cells 69 can be formed by drawing a genetic modification solution into pump 60 and pumping it into either reservoir 42 or 44 containing blood component cells. These cells 69 can be formed from larger cellular components 43 or from cells already modified by a cell concentration process or cell preparation step. The cells 69 are then drawn into pump 60. As shown in FIG. 15, the prepared cells are pumped from pump 60 through line L14, valve V9 to line L1, valves V15 and V16 to line L25, and then to inlet 74a of gene transfer module 74. This module 74 can be used to introduce a payload using electroporation, mechanoporation, or other flow-through transfection techniques. The modified cells 70 exit module 74 at outlet 74b and pass through line L32 and line L23 to line L27 via valves V18 and V19.

[0078] The gene transfer step can be performed without passing through the gene transfer module 74. Instead, a gene solution can simply be added (to the large cellular component 43) during the cell preparation step described above. For example, lipid nanoparticles can be incubated with the cells and then delivered with a payload. Optionally, a cell selection step can also be performed. Cells 72 prepared with antibodies, beads, or other solutions capable of distinguishing cells based on surface markers or phenotype can be sent to the selection chamber 76. Similar to the process shown in Figures 13 and 14, cells 72 can be formed by drawing a cell identification solution into the pump 60 and pumping it into either the reservoir 42 or 44 containing blood component cells. These cells 72 can be formed from the large cellular component 43, or from cells already modified by the cell concentration step, cell preparation step, or gene transfer step. The cells 72 are then drawn into the pump 60. 16, cells 72 are pumped from pump 60 through line L14, valve V9 to line L1, valves V15, V16, and V17 to line L26, and through inlet 76a into cell selection module 76. Cells can be positively or negatively separated, and the target cell fraction can be directed to an appropriate reservoir or output vessel.

[0079] Although various cell modification steps have been described above, not all steps in any given procedure must be performed on the collected blood cells, and selection can be made based on the desired cell composition requirements. Cells can be modified in at least one step, or in a combination of all or some of the steps. The steps do not need to be performed in a specific order, and each can be performed multiple times. The most important point is that the cells are separated and modified within the same procedure and system.

[0080] After the cells have been modified by at least one of the cell modification modules described above, they can be sent to an administration / sample container 50 (see FIG. 8 ) or, optionally, returned to the donor / patient via the fluid flow circuit. If sent to container 50, the container can be removed from the circuit. The modified cells can be returned to the donor / patient by returning them through line L1 of processing section 25 of fluid flow circuit 10 to the near-patient section 22. Various pumps and valves can be used and operated to return the fluid to the patient. The near-patient section 22 of the fluid flow circuit can initiate and control the flow of fluid back to the patient.

[0081] Several factors may be considered to determine whether cells are harvested for administration or re-infused into the patient, including guidelines imposed by regulatory agencies (e.g., FDA), the required processing time and whether the patient can remain connected during the processing time (if processing takes several hours, the patient may not need to be connected during that time), whether extensive release testing is required before re-infusion, or whether the cells require additional processing (e.g., culturing / expansion, offline administration, etc.). [Example]

[0082] The following collection and processing workflow can be implemented using a combination of the aforementioned conditions and processing procedures. Using a fluid flow circuit, the system collects a target volume of anticoagulated whole blood from a patient into reservoir 40 based on a peripheral blood pre-count of target cells. The anticoagulated whole blood is separated in a separation module, and white blood cells are separated into reservoir 42. The white blood cells are concentrated 25-fold through concentration module 72, which may include multiple passes. A biotinylated antibody additive is introduced into the system, and the white blood cells are incubated with a biotinylated antibody in reservoir 44, which is used to label all cells except CD3-positive cells. Streptavidin-coated magnetic particles are then introduced into the system and reservoir 44, and the mixture is incubated. The mixture is transferred to selection module 76, where the labeled cells are bound within the chamber. Unlabeled, unbound target CD3-positive cells are removed from the selection chamber into a reservoir, such as reservoir 42 or 44. The cells are sent to enrichment module 72, where the isolated CD3-positive cells are enriched 10-fold. A genetic modification solution (e.g., mRNA, CRISPR-Cas9, transposon / transposase) is introduced into the system and into a holding reservoir containing the CD3-positive cells. The cells pass through gene transfer module 74 and are transferred to another reservoir. These cells may be washed in separation module 62 with a buffer. The cells again pass through cell enrichment module 72 and are enriched to a target concentration for reinfusion. The enriched cells may be transferred to a dispensing container or reinfused into the patient.

[0083] Thus, improved blood component processing methods and systems have been disclosed. Advantageously, the modular fluid flow circuits described herein include cell separation and cell modification components to produce separated and modified cells in a single processing step within a single modular system. The above description is illustrative and exemplary and is not intended to limit the scope of the invention to the specific methods, systems, devices, or apparatus described herein, except as expressly limited thereto.

[0084] [Aspect] Aspect 1: A fluid flow circuit for use in a blood processing system, comprising: a separation module; at least one pump; at least one fluid reservoir for holding fluid during blood processing; a valve system; at least one fluid supply container; at least one cell modification module; a blood source access device; and a plurality of conduits fluidly connecting the components of the fluid flow circuit.

[0085] Embodiment 2 The fluid flow circuit of embodiment 1, wherein the fluid flow circuit further comprises a concentration module.

[0086] Embodiment 3. The fluid flow circuit of embodiment 1 or embodiment 2, wherein at least one cell modification module comprises a cell selection module.

[0087] Embodiment 4 The fluid flow circuit of any one of embodiments 1 to 3, wherein at least one fluid supply vessel is a buffer vessel.

[0088] Embodiment 5 The fluid flow circuit of any one of embodiments 1 to 4, wherein at least one fluid supply vessel is a solution vessel.

[0089] Embodiment 6 The fluid flow circuit of any one of embodiments 1 to 5, wherein the separation module is configured to separate blood components based on size.

[0090] Embodiment 7 The fluid flow circuit of any one of embodiments 1 to 6, wherein the at least one pump comprises a plurality of pneumatic syringe pumps.

[0091] Embodiment 8. A blood processing system comprising: the fluid flow circuit of any one of embodiments 1 to 7; and a controller configured and / or programmed to control the operation of the fluid flow circuit.

[0092] Embodiment 9 The blood processing system of embodiment 8, wherein the blood processing system is a modular system.

[0093] Aspect 10: A blood processing system as described in Aspect 8, wherein the control device is configured to operate at least one pump and valve system to transport blood from the blood source access device to the fluid flow circuit, to separate the blood using a separation module to separate the blood into two or more cellular components, and to modify at least one cellular component of the blood using at least one cellular modification module to produce at least one modified cellular component.

[0094] Embodiment 11. The blood processing system of embodiment 10, wherein the controller is further configured to operate at least one pump and valve system to deliver a solution to the at least one cellular component.

[0095] Embodiment 12. The blood processing system of embodiment 10, wherein the controller is configured to operate the at least one pump and valve system to deliver the at least one cellular component through the concentration module.

[0096] Aspect 13: The blood processing system of aspect 10, wherein the control device is configured to perform modification of at least one cellular component of the blood by operating at least one pump and valve system to deliver at least one cellular component through the gene transfer module.

[0097] Aspect 14. The blood processing system of aspect 10, wherein the controller is configured to perform modification of at least one cellular component of the blood by operating at least one pump and valve system to deliver the at least one cellular component through the cell selection module.

[0098] Embodiment 15. The blood processing system of embodiment 10, wherein the separation module is configured to separate the blood into two or more components based on the size of the two or more components.

[0099] Embodiment 16: The blood processing system of embodiment 15, wherein the size is a diameter of the cell.

[0100] Embodiment 17 The blood processing system of embodiment 10, wherein the cellular component comprises at least one of white blood cells, red blood cells, and platelets.

[0101] Embodiment 18. The blood processing system of embodiment 10, wherein the controller is further configured to operate the at least one pump and valve system to collect the at least one modified cellular component in the container.

[0102] Embodiment 19. The blood processing system of embodiment 10, wherein the blood from the blood source access device is drawn directly from the patient.

[0103] Embodiment 20. The blood processing system of embodiment 19, wherein the controller is further configured to operate at least one pump and valve system to initiate reinfusion of the at least one modified cellular component into the patient.

Claims

1. 1. A fluid flow circuit for use in a blood processing system, comprising: a separation module; at least one pump; at least one fluid reservoir for holding fluid during a blood processing procedure; a valve system; at least one fluid supply container; at least one cell modification module; a blood source access device; a plurality of conduits fluidly connecting components of said fluid flow circuit.

2. 10. The fluid flow circuit of claim 1, wherein the fluid flow circuit further comprises a concentration module.

3. 3. The fluid flow circuit of claim 1 or claim 2, wherein the at least one cell modification module comprises a cell selection module.

4. 4. A fluid flow circuit according to any one of claims 1 to 3, wherein the at least one fluid supply vessel is a buffer vessel.

5. 5. A fluid flow circuit according to any one of claims 1 to 4, wherein the at least one fluid supply vessel is a solution vessel.

6. 6. The fluid flow circuit of claim 1, wherein the separation module is configured to separate blood components based on size.

7. 7. The fluid flow circuit of claim 1, wherein the at least one pump comprises a plurality of pneumatic syringe pumps.

8. A fluid flow circuit according to any one of claims 1 to 7; a controller configured and / or programmed to control operation of said fluid flow circuit.

9. 10. The blood processing system of claim 8, wherein the blood processing system is a modular system.

10. The control device operating the at least one pump and the valve system to transport blood from the blood source access device to the fluid flow circuit; Separating the blood using the separation module to separate the blood into two or more cellular components; 10. The blood processing system of claim 8, configured to perform modification of at least one cellular component of blood using the at least one cellular modification module to produce at least one modified cellular component.

11. 11. The blood processing system of claim 10, wherein the controller is further configured to operate the at least one pump and the valve system to deliver a solution to at least one cellular component.

12. 11. The blood processing system of claim 10, wherein the controller is configured to operate the at least one pump and the valve system to deliver at least one cellular component through a concentration module.

13. 11. The blood processing system of claim 10, wherein the controller is configured to operate the at least one pump and the valve system to deliver at least one cellular component through a gene transfer module to perform modification of at least one cellular component of the blood.

14. 11. The blood processing system of claim 10, wherein the controller is configured to perform modification of at least one cellular component of blood by operating the at least one pump and the valve system to deliver at least one cellular component through a cell selection module.

15. 11. The blood processing system of claim 10, wherein the separation module is configured to separate blood into two or more components based on the size of the two or more components.

16. 16. The blood processing system of claim 15, wherein the size is a diameter of a cell.

17. 11. The blood processing system of claim 10, wherein the cellular components include at least one of white blood cells, red blood cells, and platelets.

18. 11. The blood processing system of claim 10, wherein the controller is further configured to operate the at least one pump and the valve system to collect at least one modified cellular component in a container.

19. 11. The blood processing system of claim 10, wherein blood from the blood source access device is drawn directly from a patient.

20. 20. The blood processing system of claim 19, wherein the controller is further configured to operate the at least one pump and the valve system to initiate reinfusion of the at least one modified cellular component into the patient.

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