Partially or fully automated buoyancy-assisted separation method and system

Abstract

A partially or fully automated buoyancy-assisted separation system includes and / or interacts with automated equipment. Additionally or alternatively, the system may include and / or interact with any or all of the following components: a set of buoyancy particles, a set of containers (hereinafter also referred to individually and / or collectively, equivalently as consumables), a first container management subsystem, a second container management subsystem, a user interface subsystem, and / or any other components. A partially or fully automated buoyancy-assisted separation method includes the steps of operating a set of containers and / or processing a set of material in a first container management subsystem, and operating a set of containers and / or processing a set of material in a second container management subsystem.

Description

Technical Field

[0001] The present invention generally relates to the field of biological treatment, and more specifically, to novel and useful systems and methods for partially or fully automated buoyancy-assisted separation in the field of biological treatment.

[0002] Cross-reference to Related Applications This application claims the benefit of U.S. Provisional Application No. 63 / 445,391, filed on February 14, 2023, the entire contents of which are incorporated herein by reference.

Brief Description of the Drawings

[0003] [Figure 1] FIG. 1 is a schematic diagram of a partially or fully automated buoyancy-assisted separation system. [Figure 2] FIG. 2 is a schematic diagram of a partially or fully automated buoyancy-assisted separation method. [Figure 3] FIGS. 3A - 3M show one aspect of a partially or fully automated buoyancy-assisted separation system. [Figure 4] FIG. 4 shows one aspect of a container for use in partially or fully automated buoyancy-assisted separation. [Figure 5] FIG. 5 shows one aspect of a container for use in partially or fully automated buoyancy-assisted separation. [Figure 6] FIG. 6 shows a first example of the geometric characteristics of the lower and upper chambers of a container. [Figure 7] FIG. 7 shows a second example of the geometric characteristics of the lower and upper chambers of a container. [Figure 8] FIG. 8 shows a third example of the geometric characteristics of the lower and upper chambers of a container. [Figure 9] FIGS. 9A and 9B show an example of automated equipment including first and second processing subsystems. [Figure 10]Figure 10 shows one embodiment of a mixing arm for use in a second processing subsystem. [Figure 11] Figure 11 shows one embodiment of a mixing arm for use in the first processing subsystem. [Figure 12] Figures 12A to 12E show one embodiment of an automated device and the interface between the first processing subsystem of the automated device and the consumable housing. [Figure 13] Figures 13A and 13B show examples of the first container, second container, and flexible conduit for the consumables. [Figure 14] Figures 14A and 14B show examples of valves used in consumables. [Figure 15] Figures 15A and 15B show an example of a consumable with a housing. [Figure 16] Figure 16 shows one embodiment of automated equipment. [Modes for carrying out the invention]

[0004] The following description of preferred embodiments of the present invention is not intended to limit the invention to those preferred embodiments, but rather to enable those skilled in the art to manufacture and use the invention.

[0005] 1. Overview As shown in Figure 1, a partially or fully automated buoyancy-assisted separation system 100 includes and / or interacts with an automated device 110. Additionally or alternatively, the system may include and / or interact with any or all of the following components: a set of buoyancy particles 160, a set of containers 150 (also referred to herein, individually and / or collectively, equivalently as consumables), a first container management subsystem 120 (also referred herein equivalently as a first processing subsystem and / or a first mixing subsystem), a second container management subsystem 130 (also referred herein equivalently as a second processing subsystem and / or a second mixing subsystem), a user interface subsystem 140, and / or any other components. In addition or alternatively, System 100 may include and / or interact with any or all of the components described in any or all of U.S. applications No. 16 / 004,874 filed June 11, 2018, No. 14 / 969,446 filed December 15, 2015, No. 17 / 679,688 filed February 24, 2022, No. 17 / 896,800 filed August 26, 2022, and No. 18 / 114,130 filed February 24, 2023. Each of those applications is incorporated herein by reference in its entirety.

[0006] As shown in Figure 2, a partially or fully automated buoyancy-assisted separation method 200 includes a step S300 in which a set of containers is operated and / or a set of materials is processed in a first container management subsystem, and a step S500 in which a set of containers is operated and / or a set of materials is processed in a second container management subsystem. Additionally or alternatively, the method 200 may include any or all of the following: a step S100 in which a set of materials is received in a set of containers; a step S200 in which a set of containers is received in a first container management subsystem; a step S400 in which a set of containers is received in a second container management subsystem; a step S600 in which a set of containers and / or a set of materials is removed from automated equipment; a step S600 in which a set of containers is received in another container management subsystem; a step S7 in which containers are separated and / or combined; and / or other appropriate processes.

[0007] Furthermore, or alternatively, this method may include, and / or interact with, any or all of the processes described in U.S. Patent Application No. 16 / 004,874 filed June 11, 2018, U.S. Patent Application No. 14 / 969,446 filed December 15, 2015, U.S. Patent Application No. 17 / 679,688 filed February 24, 2022, U.S. Patent Application No. 17 / 896,800 filed August 26, 2022, and U.S. Patent Application No. 18 / 114,130 filed February 24, 2023 (each of those applications is incorporated herein by reference in its entirety). Method 200 may be carried out using the system described above and / or any other suitable system.

[0008] 2. Advantages Partially or fully automated buoyancy-assisted separation systems and methods can offer several advantages compared to current systems and methods.

[0009] In a first aspect, the technology provides users with the advantage of enabling the partial or complete automation of any or all of the processes conventionally associated with particle sorting and / or separation applications (e.g., cell separation, cell therapy, cell activation, cell enrichment chemical separation, cell isolation, nucleic acid extraction, sample purification, dead cell or other particle removal, sample preparation, RNA extraction, Leukopak processing, etc.). This makes it possible to achieve any or all of the following advantages: improved efficiency in performing such processes (e.g., ensuring the availability of time-constrained materials), increased particle volume that can be processed, improved accuracy and / or yield associated with such processes, and / or other advantages. For example, the system and / or method is faster than conventional (e.g., manual) methods, gentler on cells or other target substances, more scalable (e.g., able to process hundreds of milliliters of material at once, or more than one liter at once), more versatile (e.g., supporting a wide range of starting materials, applications, and / or kits), higher yield (e.g., able to isolate more ready cells for activation, expansion, cell culture, molecular assays, etc.), and easier to use (e.g., semi-automatic or fully automated sample processing, minimal training required, manual...). One or all of the following can be obtained (compared to conventional systems and / or methods): reduced errors due to the process, time efficiency (e.g., isolating cells from one or more samples simultaneously with processing times of less than one hour per process, less than 90 minutes, 45 to 90 minutes, less than 45 minutes, 20 to 50 minutes, or processing times between any and / or other values), compliance (e.g., meeting the requirements for use in research and / or processing and manufacturing, such as gene therapy and cell therapy), and / or other advantages.

[0010] In a series of examples, automated equipment and associated methods of use enable the complete or partial automation of any or all of the following processes: adding and removing substances, mixing substances, separating substances, processing substances, collecting substances, and / or other processes.

[0011] In a second embodiment, in addition to or in lieu of the first embodiment, the technology offers the advantage of promoting and / or ensuring high sterility of any or all of the substances used in the system and / or method, for example, by facilitating the introduction of a closed system.

[0012] In a series of examples, automated equipment is configured to include a set of access ports that allow substances to be added to and / or removed from their containers in a sterile manner within a closed system. In a particular example (for example, as shown in Figures 15A and 15B), the containers of consumables are connected to tubes (for example, in both containers) or other conduits, thereby allowing substances to be added to and / or removed from the containers (for example, via access ports in a closed system manner).

[0013] Furthermore, automated equipment and / or associated methods of use can be configured to interact with other closed system components and / or processes, thereby maintaining the sterility of the closed system throughout the entire material handling process.

[0014] In a third aspect, in addition to or instead of the foregoing, the technology provides the advantage of enabling and / or optimizing any or all of the processes performed by automated equipment by using buoyant particles. For this purpose, the technology can be configured to optimize (e.g., maximize) the collection of buoyant particles and their binding material, minimize the collection of non-buoyant particles and / or non-target material, and / or provide other advantages.

[0015] In one example, a set of containers and / or a related mixing setup (e.g., within a set of a second container management subsystem) is optimized by a set of specific geometric characteristics, a set of specific quantity characteristics, a set of specific mixing protocols and / or mixing characteristics (e.g., angle of mixing, speed of mixing, etc.), and / or any or all of other characteristics for use in combination with buoyant particles.

[0016] In additional or alternative examples, the system and / or method is configured to prevent and / or minimize the adhesion of buoyant particles to the inner surfaces of the containers and / or conduits and / or other materials they may contact. For example, in a preferred embodiment of the system, any or all of the containers are shaped (e.g., having a specific pitch, bulb shape, collective hourglass shape, etc.), sized (e.g., having a set diameter, height, width, and having a minimum diameter), structured (e.g., minimizing friction material, smoothing the inner surface, coating the inner surface), and / or configured in other ways so as to maximize the amount of buoyant particles that can ultimately be collected (e.g., with respect to the target substance).

[0017] In a specific example, for instance, the dimensions of each container (e.g., length, width, height, maximum dimension, etc.) are less than 7 inches, and the container (optionally with a housing) fits within a 7 - inch cube space. Additionally or alternatively, the container can be sized to fit within a cube having dimensions of 3 - 12 inches, sized to fit within a non - cubic volume, and / or other appropriate sizes. Thereby, for example, the automated equipment can be configured to be used as a desktop device. Additionally or alternatively, a set of containers can be sized as described above, the entire consumable (e.g., the container with a housing and conduits) can be sized as described above, and / or the component can have any other appropriate dimensions.

[0018] The containers may be separate, may be connected (e.g., via conduits), may be part of a single component / piece (e.g., an hourglass-shaped single container defining multiple chambers), and / or may be arranged in other ways.

[0019] In certain series of examples (e.g., as shown in FIGS. 6 - 8, FIGS. 13A and 13B, FIGS. 15A and 15B, etc.), one or both of the containers are shaped to have an inclined shape (equivalently also referred to as pitch herein) where the diameter decreases as it approaches the other container (e.g., the diameter of the upper container decreases as it approaches the lower container, the diameter of the lower container decreases as it approaches the upper container, and the diameter is minimized where closest to the flexible conduit connecting the two containers). This can serve to guide buoyant particles from one container to the other (e.g., from the lower container to the upper container), minimize the area of the inner surface to which the buoyant particles may adhere (e.g., during transport, mixing, etc.), generate a bulk volume flow along the inner surface during mixing and / or increase contact (e.g., wipe / remove buoyant particles from the inner surface and guide them into the flexible in the flexible conduit), and / or improve the collection of buoyant particles in other ways. The containers may have the same shape as each other, different shapes from each other (e.g., as shown in FIGS. 13A and 13B, the lower container has a greater inclination than the upper container), symmetric shapes (e.g., as shown in FIG. 7, FIGS. 13A and 13B, FIGS. 15A and 15B, having a symmetric shape about an axis passing through the center of the flexible conduit), asymmetric shapes (e.g., as shown in FIG. 6 and / or FIG. 8, having an asymmetric shape about an axis passing through the center of the flexible conduit), and / or other shapes. Additionally or alternatively, the containers can have the same, similar and / or different sizes from each other, can have the same, similar and / or different materials from each other, and / or can have other suitable configurations.

[0020] In a fourth aspect, in addition to or instead of the foregoing, the technology provides the advantage of optimizing the results associated with the use of automated equipment, which can, for example, prevent consumables from being processed while they are in an suboptimal and / or unsuitable state. This is preferably achieved by ensuring that a valve is in a specific state to enable coupling of the consumable with the automated equipment, but additionally or alternatively, it can also be achieved by preventing the execution of a protocol unless the valve is in a specific configuration, and / or by operating the automated equipment in other ways to improve the results.

[0021] For example, a consumable can be physically connected to a pre-separation component of the automated equipment (e.g., a first mixed subsystem) only when the valve is in a closed configuration, and can be physically connected to a separated component of the automated equipment (e.g., a second mixed subsystem) only when the valve is in an open configuration. This is preferably made possible by arranging a set of notches that move in and out depending on the valve configuration (e.g., as shown in Figures 14A and 14B), but alternatively, it can be adequately made possible in other ways.

[0022] In another example, a valve may be configured to open automatically when a specific set of conditions (e.g., multiples of gravitational acceleration, temporal conditions, etc.) related to a separation protocol (e.g., negative selection, positive selection, human T cell negative selection, human T cell positive selection, etc.) are met, thereby allowing the valve to open naturally at the appropriate time. For example, in a specific example, the valve may include a spring-actuated mechanism that opens the conduit between containers when a certain gravitational acceleration acts during the separation process.

[0023] Additionally or alternatively, this system and method may also provide any other advantages.

[0024] 3. System As shown in Figure 1, a partially or fully automated buoyancy-assisted separation system 100 includes and / or interacts with an automated device 110. Additionally or alternatively, the system may include and / or interact with any or all of the following components: a set of buoyancy particles 160, a set of containers 150 (also referred to herein, individually and / or collectively, equivalently as consumables), a first container management subsystem 120 (also referred herein equivalently as a first processing subsystem and / or a first mixing subsystem), a second container management subsystem 130 (also referred herein equivalently as a second processing subsystem and / or a second mixing subsystem), a user interface subsystem 140, and / or any other components.

[0025] This system functions to facilitate, and more preferably optimize, one or more operations related to sample processing through automation and the use of buoyant particles. In a preferred embodiment, for example, the system functions to perform buoyant separation of particles in a sample through automation carried out using automated equipment. Furthermore, the system can function to improve the efficiency related to any or all processes, increase the volume and / or yield related to any or all processes (compared to, for example, performing the processes manually), and / or perform other appropriate functions.

[0026] 3.1 System: Automation equipment 110 System 100 includes and / or interacts with an automated device 110, which functions to enable any or all of the following: customization of workflows and / or protocols; increased batch sizes that can be processed by a single protocol (e.g., multiple containers, multiple Leukopaks, etc.); assurance of sterility through closed-system workflows; faster workflows and / or improved yields compared to conventional and / or manual systems; easier preparation and / or recovery of separated materials (e.g., by performing non-magnetic processes); and / or other outcomes.

[0027] The automated device 110 can optionally be configured to communicate (e.g., in a modular manner) with other devices, storage containers, and / or parts where sample material may be supplied and / or processed. This can, for example, function to maintain the sterility of the sample (e.g., through the introduction of a complete closed system between devices).

[0028] The automated instrument 110 is preferably configured to automate (e.g., partially automated, fully automated) at least a particle separation process (e.g., cell separation, isolation of a set of target substances from the rest of the sample), and more preferably configured to enable high-performance positive selection (e.g., cell activation) applications, high-performance negative selection applications, any other applications, and / or any combination of these applications.

[0029] The automated equipment 110 is more preferably configured to perform aseptic closed-system processing of the substance handled by the automated equipment, which is made possible by one or more components of the automated equipment (e.g., a Luer lock that allows aseptic addition and removal of the substance to and from the container), a manufacturing process used to manufacture the automated equipment (e.g., aseptic welding), and / or any other functions of the automated equipment.

[0030] In a preferred embodiment, the automated equipment includes, but is not limited to, a set of container management subsystems (described later), for example, a first container management subsystem 120 and / or a second container management subsystem 130. The container management subsystems preferably function to operate (e.g., move, translate, rotate, actuate, heat, cool) one or more containers of the system 100 (e.g., individually or collectively), and the system may function to mix the contents of one or more containers (e.g., by combined actions according to a protocol), facilitate the movement of buoyancy particles to separate material bound to buoyancy particles from the remaining bulk volume, add and / or remove the contents of one or more containers, heat, cool and / or otherwise process the material in the containers and / or otherwise process the material in the containers.

[0031] The automated equipment may include, but is not limited to, any number of components configured to enable any or all of the functions described above and / or described below, such as one or more actuators (e.g., motors, robotic arms, electrically actuated components, pneumatically actuated components, hydraulically actuated components, etc.), power supplies, fixtures (e.g., container holders), and / or other components.

[0032] Additionally or alternatively, automated equipment may include and / or interact with any other components.

[0033] 3.2 System: Set of 150 containers The system preferably includes and / or is combined with a set of containers 150 (for example, as shown in Figure 1, Figures 3E-3G and 3K-3M, Figure 4, Figure 5, Figures 13A and 13B, Figures 15A and 15B), the set of containers serving to contain the material to be processed by automated equipment (for example, in Method 200). The set of containers can further serve to enable and / or facilitate any or all of the following outcomes: sterilization of the material (e.g., in a closed system), separation of the material in a sample (e.g., separation by separable chambers, separation by sealing the chambers, separation by valves separating the chambers, etc.), optimized collection and / or distribution of target particles in a portion of the container (e.g., by the geometric properties of the containers, the material selection of the containers, etc.), and / or other outcomes.

[0034] A set of containers is also referred to herein collectively as a consumable (for example, together with other components such as housings, connecting conduits, valves, etc.). Consumables are single-use, reusable, and / or can be used a suitable number of times.

[0035] In a preferred series of embodiments, the consumables include an upper container, a lower container, conduits connecting the upper and lower containers, housings, valves, and / or other components.

[0036] Each container preferably comprises one or more chambers (also referred to herein as equivalently as cavities) that function to contain the contents of the container, but additionally or alternatively, other configurations are possible. In a preferred set of embodiments, each container comprises a single chamber. In alternative embodiments, one or more containers comprise multiple chambers and / or are configured to be separable into multiple chambers (e.g., by opening and closing valves, heat sealing, mechanical separation, etc.).

[0037] Each container is preferably a rigid container (e.g., a rigid-walled container) (e.g., a non-deformable, non-elastic, non-flexible container), and is made of a material having any or all of the following characteristics: rigidity exceeding a preset threshold (e.g., shear modulus, Young's modulus, etc.) (e.g., Young's modulus exceeding 1 gigapascal (GPa), Young's modulus exceeding 0.5 GPa, Young's modulus exceeding 1.5 GPa, Young's modulus between 1 and 10 GPa, Young's modulus between 1 and 5 GPa, etc.), elasticity below a preset threshold, wall thickness exceeding a preset threshold (e.g., wall thickness exceeding 0.25 mm, wall thickness exceeding 0.5 mm, wall thickness exceeding 0.75 mm, wall thickness exceeding 1 mm, wall thickness exceeding 2 mm, wall thickness between 2 and 3 mm, wall thickness within the range defined by any of those values, etc.), and / or other characteristics. In a preferred series of embodiments, each container is made of plastic or other polymer material (e.g., polycarbonate (PC), polyvinyl chloride (PVC), etc.).

[0038] In alternative embodiments, one or more of the set of containers may be made of a material that is at least partially deformable (e.g., flexible, expandable, pliable, thin-walled, etc.) and thereby configured to give the containers variable volume (e.g., expandability, contractility, etc.). This includes, for example, materials with a Young's modulus below a predetermined threshold (e.g., Young's modulus less than 1 GPa, Young's modulus less than 0.75 GPa, Young's modulus less than 0.5 GPa, etc.), containers with a wall thickness below a predetermined threshold (e.g., less than 1 millimeter (mm), less than 0.5 mm, less than 0.25 mm, etc.), materials with elasticity and / or flexibility exceeding a predetermined threshold, and / or any other material. Additionally or alternatively, any or all of the set of containers may be made of rigid materials (e.g., plastics, polymers, metals, etc.), semi-rigid materials, combinations of materials (e.g., a flexible chamber coupled with a rigid chamber), and / or any other material.

[0039] In a series of embodiments, for example, the set of containers includes a set of flexible bags, the bags being expandable, deformable, separable into multiple chambers, and / or otherwise manipulable.

[0040] Additionally or alternatively, the container may have any other suitable form factor and / or be made of any suitable material.

[0041] The consumables preferably include and / or can be configured as a set of multiple parts (e.g., multiple chambers, multiple containers, etc.), where the parts are fluidically connected (e.g., in response to the opening of valves positioned between the parts during a part of the method), fluidically isolated (e.g., during a part of the method, while valves separating the parts are in a closed configuration), completely separable (e.g., separable into two separate bags after a sealing and / or separation process), and can be switched between configurations (e.g., from a fluidically isolated state to a fluidly connected state when valves are opened), and / or otherwise configured. Alternatively, any or all of the set of containers may include a single part (e.g., a single chamber).

[0042] Multiple parts may be associated with the same or different heights, the same or different volumes, the same or different shapes, the same or different materials, and / or have any other arbitrary properties.

[0043] The consumables preferably include a set of one or more conduits (e.g., tubes, cylinders, etc.) configured to fluidly connect two or more containers of a set of containers. The set of conduits can be flexible (e.g., less rigid than the containers), rigid, or any combination. In a preferred series of embodiments (e.g., as shown in Figures 13A and 13B), the consumables include a conduit (e.g., a flexible conduit, a PVC conduit, etc.) positioned between a lower container and an upper container, thereby allowing the fluid connection between the containers to be selectively allowed and blocked (e.g., as described later) via a valve. In a specific example, the conduit includes a flexible conduit made of PVC, which allows for both the deformability of the flexible conduit by the action of a valve (e.g., as described later) and the prevention of sticking and / or kinking of the inner wall of the flexible conduit.

[0044] In a preferred series of embodiments (for example, as shown in Figures 3E-3G and 3K-3M, as shown in Figure 4, and as shown in Figure 5), the container comprises at least a first container defining at least a first chamber and a second container defining at least a second chamber, wherein the second container (equivalently also referred to herein as the upper container) is positioned above the first container (equivalently referred to herein as the lower container). This allows buoyant particles to flow from the first chamber to the second chamber by buoyancy (for example, when a valve connection between the chambers is opened). This arrangement can be further configured to prevent non-buoyant particles and / or substances not bound to buoyant particles from flowing naturally into the upper chamber (equivalently, under gravity, without inverting the container). In a series of examples, the second chamber (equivalently also referred to herein as the upper chamber) contains (i.e., is pre-filled) a fluid (e.g., culture medium) before the upper chamber and the first chamber (equivalently referred to herein as the lower chamber) are fluidly connected, and this fluid (e.g., pre-filled culture medium) functions to allow and facilitate the flow of sample material into the upper chamber when the valve is opened. Additionally or alternatively, the upper chamber may be fluidless, contractible, gas-inflatable, and / or otherwise configured.

[0045] Pre-filling the second chamber also serves to expand (e.g., widen, open) the flexible conduit positioned between the first and second containers, thereby removing any kinks or adhesions between the inner walls of the flexible conduit before the separation process, allowing the buoyant material to move (e.g., float) from the lower chamber to the upper chamber.

[0046] Additionally or alternatively, the consumables may include three or more containers, a single container, multiple chambers, a single chamber, or any other suitable configuration.

[0047] In the first example (for example, as shown in Figure 3E), the container has a first chamber and a second chamber, the second chamber positioned above the first chamber, and the first and second chambers are separated by a valve and / or other separation component (e.g., a conduit, a flexible conduit, a conduit connected to a valve, etc.) (e.g., initially during a series of processes in Method 200). In one example of use of such a separation container, the first chamber initially contains the sample material (e.g., introduced via a set of access ports (e.g., a Luer lock port on a set of first container management subsystems)), and the second chamber initially contains culture medium or other fluid (e.g., pre-filled) (e.g., if the second chamber is in the form of a gas-permeable cell culture bag pre-filled with culture medium), and the first and second chambers are fluidically coupled by opening a valve during one or more processes of Method 200 (e.g., S500, after buoyancy particles have bound to the target material, etc.). Additionally or alternatively, all substances can be added to one of the set of containers (e.g., the upper container) until the separation process begins (for example, if all substances are mixed in one container before the valve between the containers is opened).

[0048] The container and / or associated chambers are preferably configured to have geometric properties configured to maximize and / or optimize the movement of buoyant particles and binding material from one chamber (e.g., a lower chamber) to another chamber (e.g., an upper chamber). This includes, for example, maximizing the proportion of buoyant particles that ultimately move from one chamber to another, minimizing the proportion of the remaining sample (e.g., non-buoyant particles and non-buoyant binding particles) that is transferred from the chamber, achieving a uniform and / or nearly uniform distribution of buoyant particles within the destination chamber (e.g., minimizing the accumulation of buoyant particles in specific sub-regions), and / or otherwise maximizing and / or optimizing this movement. Additionally or alternatively, the geometric properties may further function to minimize breakage of buoyant particles (e.g., glass buoyant particles) by gently guiding the buoyant particles (and / or preventing them from directly colliding with the inner wall of the container), and / or bring other advantages to the system and / or method.

[0049] These geometric characteristics preferably include a change in the diameter of each container, and more preferably, a change in diameter to achieve a sloping and / or curved outer shape of the container as the rigid container approaches a fixed position between containers (e.g., as it approaches a valve, flexible conduit, etc.), as shown in any or all of Figures 3E-3G, 3K-3M, 4-8, 13A-13B, and 15A-15B. The sloping can be a linear outer shape (so-called linear sloping) (e.g., Figures 13A-13B, Figure 6, etc.), a curved outer shape (e.g., Figures 7, 8, etc.), a combination of curves and straight lines, and / or other suitable configurations.

[0050] For example, in some cases, a flow of buoyant particles is formed during or after a spin process (e.g., light centrifugation by a second container management subsystem). The pitch of the inclined containers (i.e., the degree of inclination) can facilitate the induction of flow between containers (e.g., preventing the rear of buoyant particles from being left behind, preventing buoyant particles from adhering to the inner wall near the spout). Furthermore, narrowing the upper container can also serve to retain the buoyant particles (e.g., after they enter the upper container).

[0051] The angle of the slope (e.g., a slope as shown in Figure 13A) (e.g., a single angle, an average angle, or an angle aggregated in any other way) can take any suitable one or more values, such as 30–60 degrees, 45–90 degrees, 30–45 degrees, 20–60 degrees, and / or any other arbitrary range or endpoint of a range defined within any of these values.

[0052] Each container can be symmetrical about the central axis (e.g., Figures 13A and 13B), asymmetrical about the central axis (e.g., Figure 8), and / or have other shapes.

[0053] The inclination angle may differ from container to container (e.g., Figures 13A and 13B), be the same across containers, and / or be otherwise suitable. Additionally or alternatively, any other features (e.g., shape, size, etc.) may differ from container to container.

[0054] For example, in a series of embodiments (for example, as shown in Figures 3E-3G and 3K-3M, as shown in Figure 4, and as shown in Figure 5), the outer shape of the lower vessel is composed of inclined and / or curved sides extending from the upper region of the lower vessel to the lower region of the upper vessel, for example, a frustoconical and / or nearly frustoconical shape (e.g., curved sides, asymmetrical sides, asymmetrical curved sides, etc.), and the diameter decreases as the height increases, thereby guiding buoyancy particles from the lower chamber to the upper chamber (for example, through a passage connecting the chambers if a valve or other component is arranged in an open configuration).

[0055] Additionally or alternatively, the lower chamber may have one or more straight sides, inclined sides with increasing diameter along with height, and / or other suitable shapes.

[0056] In some cases (for example, as shown in Figure 5, Figure 6, and Figure 8), the consumables have an asymmetrical shape, and the slope (e.g., average slope) of one part / region (e.g., the side) of the lower container is steeper than the slope of other parts / regions (e.g., the side). This can function in a second container management subsystem (e.g., described later) to maximize the movement and / or ease of movement of buoyancy particles to the upper chamber (e.g., when the valve is open) when the container is spinning around an offset axis. For example, the steeper slope can function to guide buoyancy particles more easily from the lower container to the upper container (e.g., in the presence of centripetal forces due to the rotation of the container) during the rotational motion (e.g., spinning) of the second container management subsystem. This can prevent buoyancy particles from accumulating in the passage between chambers, allow buoyancy particles to accumulate relatively uniformly in the upper chamber, and / or otherwise optimize the movement of buoyancy particles from the lower chamber to the upper chamber. Additionally or alternatively, the connection between the lower and upper chambers can be angled (for example, non-vertically as shown in Figure 8), which can further facilitate the induction of material from the lower chamber to the upper chamber (for example, when a valve located between the chambers is opened).

[0057] In other examples (for instance, as shown in Figure 4 and Figure 8), the inclination may be symmetrical. In specific examples, the movement of particles within such a container (and / or other containers such as asymmetric containers) can be optimized and / or made more efficient through a spin protocol of a second container management subsystem, for example, by reversing the spin direction (e.g., for a portion of the total spin period).

[0058] In other examples (for example, as shown in Figures 13A and 13B), the upper and lower containers are symmetrical about a central axis, but one container (e.g., the lower container) is steeper than the other. For example, in the specific examples shown in Figures 13A and 13B, the lower container has a greater incline (i.e., a smaller angle) than the upper container, which helps to guide the buoyancy particles into the upper container and then retain them within the upper container.

[0059] One or more containers (e.g., upper containers) are preferably configured to have a rounded shape, and more preferably to have a spherical shape (e.g., a circular profile) and / or an oval shape (elliptical profile). This serves to generate maximum gravity at the location of the channel between the lower and upper chambers, thereby facilitating the movement of buoyancy particles from the lower chamber to the upper chamber (e.g., during the spin of the container in the second container management subsystem). Additionally or alternatively, the lower chamber may have such a shape, the upper chamber may have any suitable shape (e.g., an asymmetric slope region, a symmetric slope, etc., defined for the lower chamber), and / or the lower chamber may have any other suitable shape.

[0060] The consumables more preferably include a housing (as shown, for example, in Figures 15A and 15B) which functions to partially or completely enclose the container throughout part or all of Method 200. Additionally or alternatively, the housing may function to support and / or define one or more valves (for example, as described below), enable an interface (e.g., coupling, optimal coupling, etc.) between one or more containers and automated equipment, protect one or more flexible conduits (and / or flexible containers in embodiments such as those using bag-based containers), and / or provide any other arbitrary function.

[0061] In a preferred series of embodiments (for example, as shown in Figures 15A and 15B), the housing includes a rigid shell that protects and enables a selective connection between the container and the automated equipment, and also allows for the operation of the container within the automated equipment.

[0062] Additionally or alternatively, the housing can have other configurations.

[0063] 3.3 System: Valves and / or conduits Consumables preferably include valves and / or other components configured to selectively provide passage of material between chambers. The valves may be passively actuated (e.g., spontaneously open during the spinning of the container by a second container management subsystem), actively actuated (e.g., actuated by an actuator or other component of an automated device opening and closing the valve based on a protocol), and / or any combination thereof.

[0064] The valve can optionally be part of the housing and / or fixed to the housing, and can further serve to ensure that the valve is in an appropriate configuration in each of the container management subsystems (e.g., closed for mixing in the first container management subsystem, open for separation in the second container management subsystem, etc.).

[0065] Additionally or alternatively, the valve can be properly configured in other ways.

[0066] The valve is preferably connected to one or more conduits (e.g., tubes, hoses, cylinders, spouts formed by containers, etc.) (for example, as described above), and the valve can selectively allow (and block) connections between containers via one or more conduits. The valve can be located outside one or more conduits (e.g., actuated on the outer wall of the conduit), inside one or more conduits (e.g., actuated on the inner wall of the conduit, in the internal space of the conduit, etc.), and / or arranged in any combination.

[0067] Alternatively, the valve can be used without a conduit. For example, in some embodiments, the valve is located inside one or more containers (e.g., inside or near the spout of one or more containers).

[0068] In a preferred series of embodiments (for example, as shown in Figures 15A and 15B), the valve is positioned outside one or more vessels and a deformable conduit connecting them, and the valve selectively blocks the flow through the flexible conduit by reducing the diameter of the flexible conduit, such as by clamping, twisting, and / or otherwise reducing the diameter of the conduit (e.g., to a value below a preset threshold, nearly zero, or zero). In one example (for example, as shown in Figures 13A and 13B, and as shown in Figures 14A and 14B), a set of valve pieces (e.g., clamps) can be rotated relative to each other (e.g., as shown in Figures 14A and 14B), and by rotating the valve pieces in a particular direction, the flexible conduit can be compressed (its diameter reduced), and by rotating the valve pieces in the opposite direction, the conduit can have its full diameter. Additionally or alternatively, the valve pieces can be operated to restrict or restrict and release flow by translating them (e.g., pressing them against each other).

[0069] The valve may optionally include one or more levers and / or tabs (for example, as shown in Figures 14A and 14B), which may change the operation of the valve (for example, by moving the lever clockwise or counterclockwise, the radius of the internal opening of the lever may be changed), allow for selective coupling of the housing and automated equipment, indicate the operating state of the valve, and / or perform appropriate functions in other ways.

[0070] In the examples shown in Figures 14A and 14B, the valve includes an external valve having multiple valve pieces, and by rotating these pieces relative to each other, the diameter of the opening formed by these pieces changes, thereby either trapping (e.g., restricting, closing) or releasing (e.g., expanding to a fully widened diameter) a conduit placed within the opening. In the specific example shown in Figure 14B, the distance from the central axis (passing through the center of the conduit) to the first end of the curved opening ("A") is different from the distance from the central axis to the second end of the curved opening ("B"), and this difference in distance changes the size (e.g., diameter) of the opening formed between the pieces. For example, as shown in Figure 14B, since "A" is longer than "B", rotating the first valve piece counterclockwise pushes a portion of the second valve piece inward, trapping and closing the conduit (e.g., as shown in Figure 13B). Furthermore, changes in the position of the rotating lever can also be utilized for selective connection with automated equipment (e.g., as described above and / or later).

[0071] In an alternative example, the valve is a single piece, the valve pieces can move in parallel relative to each other, the valve pieces can change height when rotated, and / or the valve can function properly in other ways.

[0072] In some embodiments, a lever needs to be configured in a specific way to connect (e.g., lock) the housing / consumable to the automated equipment, thereby ensuring that the valve is in the appropriate state for a particular process of the automated equipment. This can function as a fail-safe mechanism (e.g., the user will not attempt a separation process with the valve closed, nor will the user attempt a mixing process with the valve open).

[0073] In certain cases (for example, as shown in Figures 14A and 14B), the lever rotates with the valve, and when the valve is in a first configuration (e.g., open, closed, etc.), the lever protrudes from a set of slots in the housing, and when the valve is in a second configuration (e.g., closed, open, etc.), it does not protrude from the set of slots (e.g., retracts). This allows the container management subsystem of the automated equipment to be configured to accept the housing based on the state of this lever. For example, if the lever retracts when the valve is closed, the first container management subsystem can be configured to have a projection that engages into a slot, so that the housing can be coupled with the first container management subsystem only when the valve is in a closed configuration (e.g., as shown in Figure 11, as shown in the “click” in Figure 12E). Also, if the lever protrudes when the valve is open, the second container management subsystem can be configured to have a set of openings (e.g., as shown in Figure 10), so that the second container management subsystem can accept and connect to the housing only when the valve is in an open configuration. Alternatively, the container management subsystem could be configured in reverse, the levers in a different way, and / or the system could be configured appropriately in a different way.

[0074] In alternative embodiments, the system may include valves configured to open on their own during a particular process and / or when a particular set of conditions (e.g., force, a particular multiple of gravity (G), velocity, direction, etc.) is experienced. The valves are preferably configured to open during or immediately before the separation process (e.g., during processing by a second container management subsystem), but additionally or alternatively, they may be configured to close at one or more points in time and / or conditions (e.g., after buoyancy particles have moved to the upper container, after separation is complete, based on temporal conditions such as exceeding a threshold time, after being actuated by centrifugal force, based on the angle threshold of the swing bucket being met in the second container management subsystem), or to open and close based on the automatic operation and / or activation of tools in automated equipment (e.g., robotic arms, magnets, etc.), and / or one or more valves may be actuated appropriately in other ways.

[0075] In one example, a valve is positioned within a conduit and / or within a portion of one or more rigid containers (e.g., within a spout), and can transition from a closed configuration to an open configuration when a threshold force (e.g., a multiple of gravitational acceleration, 1 to 20 times gravitational acceleration, 5 to 30 times gravitational acceleration, etc.) acts on the valve (e.g., during swing bucket centrifugation in a second container management subsystem). In a specific example, the valve includes a spring and a mass, where the threshold force causes the mass to push down the spring, opening the valve and allowing fluid flow. Additionally or alternatively, the valve may include any other suitable components, such as a duckbill component (e.g., a duckbill valve), a stopper, a float, and / or other components.

[0076] Valves can be operated manually (e.g., by a user), automatically (e.g., by automated equipment), controlled by other means, and / or by any combination of these methods.

[0077] Additionally or alternatively, the valve can be adequately configured in other ways.

[0078] 3.4 System: Container Management Subsystem 120, 130 The automated equipment preferably includes a set of container management subsystems that have the function of operating a set of containers (for example, according to a protocol, according to a protocol specific to the set of substances), the operation of the containers functions to perform and / or facilitate the processing of the substances (e.g., separation, isolation, collection, mixing, etc.). The set of container management subsystems preferably includes multiple container management subsystems, which may have the same function, different function, and / or any combination of functions. Alternatively, the set of container management subsystems may also include a single container management subsystem.

[0079] Each container management subsystem preferably acts on all containers of the consumables (e.g., manipulating, moving, or rotating the containers). Alternatively, any or all of the container management subsystems may act on a single container or a subset of containers, or any other part of the system.

[0080] In a preferred set of embodiments (for example, as shown in Figures 3A–3C and 3H–3J), a set of container management subsystems includes a first container management subsystem and a second container management subsystem, each of which is preferably configured to operate a set of containers (so-called consumables) and / or housings (e.g., automatically or semi-automatically) (e.g., to perform translation, rotation, connection, adding and / or removing material, etc.). Additionally or alternatively, the set of container management subsystems may include additional container management subsystems, subsets of those container management subsystems, and / or other container management components.

[0081] The first container management subsystem is preferably configured to perform any or all of the following functions: adding and / or removing substances, mixing substances (e.g., rotational mixing, end-over-end mixing, etc.), transferring containers from the first container management subsystem to the second container management subsystem, and / or any other functions.

[0082] The first container management subsystem (and / or other container management subsystems) preferably operates according to a protocol (e.g., a user-customizable protocol, a user-configurable protocol, a dynamically determined and / or adjusted protocol), such as a mixing protocol (e.g., end-over-end mixing, modified end-over-end mixing). The protocol may define any number of parameters and parameter values, including, but is not limited to, speed (e.g., rotational speed), angle (e.g., range of angles over which the mixing plate rotates, maximum rotation angle, start angle and / or end angle and / or intermediate angle), acceleration (e.g., rotational acceleration, to define vortex, to define vibration, etc.), temporal parameters (e.g., pause, timing of stop, to define movement to a specific angle before stopping, etc.), and / or other arbitrary parameters.

[0083] In some embodiments, the concentration of cells (or other target substances) and / or the total amount of the substance to be separated (e.g., bulk volume, sample volume) may affect any or all of the parameters. For example, if a small amount of sample (e.g., target substance, bulk volume) is present, a shorter or lower intensity mixing protocol (e.g., slower rate, narrower angular range) may be implemented to prevent the small amount of sample from covering the inside of the container and preventing optimal separation.

[0084] In a preferred series of embodiments (for example, as shown in Figures 12A to 12E, as shown in Figure 11), the first container management subsystem is configured to mix the contents of one or more containers of consumables (for example, by end-over-end mixing, modified end-over-end mixing, etc.) by the rotation of the consumables. Furthermore, the first container management subsystem can transfer the contents of the containers or process them in other ways (for example, by heating, cooling, adding substances, extracting substances, etc.). Therefore, the first container management subsystem preferably includes a fixture (e.g., a holder) that holds (e.g., fixes) the consumables (e.g., via a housing, via a tab in the housing, via an opening in the housing) and rotates them (e.g., perpendicular to the central axis of the consumables) according to a protocol (e.g., perpendicular to the central axis of the consumables) (e.g., via a motor with an encoder that drives a mixing shaft and plate, as shown in Figure 11). The holder may optionally include or specify any number of features, such as an opening (for coupling with consumables), slots, tabs, prongs, projections, clips, screws or other mechanisms for attaching the holder to the rest of the automated equipment (for example, the slot in Figure 10 may allow the holder to be coupled to the automated equipment and / or maintain the relative orientation of the holder to the equipment), and / or other arbitrary features. Additionally or alternatively, the first container management subsystem may perform other types of mixing (e.g., spinning around a central axis, centrifugation, etc.) and / or other arbitrary processing.

[0085] In one example, a first container management subsystem is configured to perform corrective end-over-end mixing of the contents of one or more containers (e.g., upper container, lower container, etc.). In a particular example, the protocol includes rotating a consumable in a first direction (e.g., counterclockwise, clockwise, etc.) by a first angular amount (e.g., 360 degrees, 180-360 degrees, 270 degrees, greater than 360 degrees, less than 360 degrees, or any range between those values), stopping (e.g., instantaneously, for a predetermined period of time, etc.), and then rotating the consumable in the opposite direction by a second angular amount (e.g., the same angular amount as the first angular amount, a different angular amount, etc.), thereby effectively forming a pendulum-like mixer.

[0086] In other embodiments, the first container management subsystem includes and / or is connected to a set of cartridges (e.g., canisters as shown in Figure 3C) that house a set of containers, and the first container management subsystem includes a housing cavity that houses the cartridges and associated components (e.g., actuators, motors, wheels, tracks, etc.) for operating the set of cartridges. The containers may be empty, or some or all of the material may be pre-filled (e.g., the upper chamber may be pre-filled with culture medium), and / or may be configured in other ways when housed in the first container management subsystem (e.g., in the early stages of the protocol). The cartridge (e.g., a cylindrical canister) is preferably configured to receive a set of containers, hold the set of containers (e.g., in an optimal configuration), rotate the set of containers (e.g., to perform end-over-end mixing to perform mixing of substances in a chamber and / or containers), move the set of containers (e.g., between the positions of automated equipment), process the substances in the set of containers in other ways (e.g., heating, cooling, etc.), and / or perform other processes. For example, in one example, the first container management subsystem includes a set of cylindrical canisters configured to rotate (e.g., automatically, semi-automatically, according to a rotation protocol) thereby performing end-over-end mixing of the contents of the containers. This can serve, for example, to facilitate the binding of buoyancy particles with associated target substances (e.g., in a lower chamber). Additionally or alternatively, the first container management subsystem may function to move to and / or be located within the second container management subsystem (e.g., as shown in Figures 3H-3J) for additional processing.

[0087] The chambers of the containers are preferably fluidically separated from each other (i.e., the valves are closed) while the consumables are in the first container management subsystem (e.g., within the containment opening shown in Figure 3C), but alternatively, they may be fluidically connected for part or all of this period. In one example, the chambers are fluidly connected only after the canister of the first container management subsystem has moved to the second container management subsystem (e.g., after the valves have been opened during mixing).

[0088] The second container management subsystem is preferably configured to enable and / or optimize the movement (equivalently also referred to herein as separation) of material between chambers (e.g., through valve openings), and more preferably to optimize the movement (e.g., in terms of efficiency, yield, etc.) of buoyant particles and material bound to buoyant particles from the lower chamber to the upper chamber. This is preferably enabled by spin (e.g., gentle spin, spin at a gravitational acceleration significantly smaller than that of a conventional centrifuge, 10 to 150 times the gravitational acceleration, 25 to 125 times the gravitational acceleration, 10 to 50 times the gravitational acceleration, 15 to 100 times the gravitational acceleration, less than 150 times the gravitational acceleration, less than 125 times the gravitational acceleration, less than 100 times the gravitational acceleration, etc.), but additionally or alternatively, it may also be enabled by other suitable spin components and / or protocols and / or parameters.

[0089] The second container management subsystem is preferably operated according to a protocol (e.g., the protocol described above for the first container management subsystem) that can control any or all parameters of the automated equipment. These include speed, angle, acceleration, time, direction of rotation (e.g., clockwise, counterclockwise, etc.), combination of parameters (e.g., move at a first speed for a first period, move at a second speed for a second period, etc.), and / or other parameters (e.g., described above). One or more protocols may be pre-configured (e.g., selected by the user), dynamically determined and / or adjusted, determined based on user input (e.g., in a user interface), and / or any combination thereof.

[0090] For example, one or more protocols of a spin subsystem (e.g., for cell separation) can be configured to specify a rotation speed below a preset threshold, thereby promoting and protecting cell health.

[0091] The second container management subsystem is preferably configured to enable and / or facilitate (e.g., facilitate, allow, optimize) the separation (e.g., longitudinal separation, suspension separation, etc.) of buoyant particles and bound target material from some or all of the rest of the bulk volume. In a preferred embodiment, the second container management subsystem does this by spinning the consumables (e.g., centrifugal separation). Additionally or alternatively, the second container management subsystem may facilitate the separation by static treatment of the consumables, other motion of the consumables (e.g., translation, rotation, shaking, etc.), other treatment of the consumables (e.g., heating, cooling, etc.), any combination of those treatments, and / or any other treatment.

[0092] In a preferred set of embodiments, for example, the second container management subsystem includes a spin subsystem (e.g., a fixed bucket mixer / centrifuge, a swing bucket mixer / centrifuge as shown in Figure 16, a centrifuge, etc.) configured to facilitate the movement of buoyant particles into the upper chamber (in addition to natural vertical movement brought about by buoyancy). Additionally or alternatively, other features may contribute to optimizing this movement, including, but are not limited to, the shape and / or other geometric features of the container (e.g., the rounded shape of the upper chamber, the asymmetric shape of the lower chamber, etc., as described above), pre-filling of the upper chamber with fluid, features of the protocol (e.g., spin velocity, spin angle, etc.), and / or other features.

[0093] In one example, the second container management subsystem includes a swing bucket centrifuge (e.g., one that reaches an angle / horizontal configuration of at least 45 degrees when spinning).

[0094] In another example, the second container management subsystem includes a fixed-angle bucket centrifuge.

[0095] The spin subsystem can optionally additionally function to open valves or other isolation components located between multiple chambers of a vessel. For example, the spin subsystem may be configured to generate the minimum force (e.g., gravitational acceleration) required to open a passively actuated valve. Additionally or alternatively, the valve may be opened by another component of the automation subsystem (e.g., an actuator, electronic component, magnetic component, etc.) and / or otherwise appropriately opened. The spin subsystem and / or other components of the automation equipment may optionally be further configured to close the valve (e.g., after buoyancy particles have moved into the upper chamber) based on the implementation of an optimized spin protocol (e.g., enabling the passive closing of the valve at an optimal timing) and / or a component (e.g., an actuator, electronic component, magnetic component, sensor, etc.) configured to close the valve.

[0096] The second container management subsystem may include any number of fasteners (e.g., holders) configured to hold / connect consumables (e.g., via housings) (e.g., as shown in Figure 10).

[0097] In a specific example, the rotating subsystem includes a bucket mixer, and the buckets can be fixed (e.g., vertically, horizontally, and fixed at a preset angle (e.g., 45 degrees, 30-60 degrees, etc.)), swing (e.g., variable angle, swinging at a variable angle up to 45 degrees relative to the vertical), and / or any combination of these.

[0098] Additionally or alternatively, a second container management subsystem may include and / or otherwise appropriately configure any other components.

[0099] 3.5 System: User Interface Subsystem 140 The automated equipment may optionally include a user interface subsystem, which may include one or all of the following: a display (e.g., for displaying information about a protocol to the user, for accepting protocol selection or other input from the user), a scanner (e.g., for scanning barcodes associated with a container or other material container configured to communicate a specific protocol performed by the automated equipment), and / or other appropriate components (e.g., a mouse, touchpad, keyboard, tube sealer, tube welder, etc.).

[0100] The system may preferably include and / or interface with any or all of the sets of buoyancy particles described in any or all of U.S. applications 16 / 004,874 filed June 11, 2018, 14 / 969,446 filed December 15, 2015, 17 / 679,688 filed February 24, 2022, 17 / 896,800 filed August 26, 2022, and 18 / 114,130 filed February 24, 2023 (each of these applications is incorporated herein by reference in its entirety). The buoyancy particles preferably feature a density lower than the density of the other particles in the set of material (and / or the average density of the bulk fluid in the sample), thereby configuring the buoyancy particles to float on the surface of the fluid in which they are immersed. More preferably, the buoyancy particles are configured to be functionalized with factors (e.g., antibodies, moieties, etc.) configured to facilitate the binding of the buoyancy particles to specific target particles of a set of substances. Additionally or alternatively, the buoyancy particles can be adequately configured in other ways.

[0101] Additionally or alternatively, system 100 may include and / or interact with any other suitable components.

[0102] 4. Method 200 As shown in Figure 2, a partially or fully automated buoyancy-assisted separation method 200 includes a step S300 in which a set of containers is operated and / or a set of materials is processed in a first container management subsystem, and a step S500 in which a set of containers is operated and / or a set of materials is processed in a second container management subsystem. Additionally or alternatively, the method 200 may include any or all of the following: a step S100 in which a set of materials is received in a set of containers; a step S200 in which a set of containers is received in a first container management subsystem; a step S400 in which a set of containers is received in a second container management subsystem; a step S600 in which a set of containers and / or a set of materials is removed from automated equipment; and / or other suitable processes.

[0103] Furthermore, or alternatively, this method may include, and / or interact with, any or all of the processes described in U.S. Patent Application No. 16 / 004,874 filed June 11, 2018, U.S. Patent Application No. 14 / 969,446 filed December 15, 2015, U.S. Patent Application No. 17 / 679,688 filed February 24, 2022, U.S. Patent Application No. 17 / 896,800 filed August 26, 2022, and U.S. Patent Application No. 18 / 114,130 filed February 24, 2023 (each of those applications is incorporated herein by reference in its entirety). Method 200 may be carried out using the system described above and / or any other suitable system.

[0104] Method 200 preferably functions to enable the separation of substances in a sample in an automated (e.g., fully automated) or semi-automated manner, but additionally or alternatively, it may perform other suitable functions.

[0105] Method 200 may optionally include step S100, in which a set of substances is received in a set of containers, which functions to receive any or all of the particles, buffers, culture media, and / or other substances to be processed in Method 200 in a sterile condition (e.g., a closed system). Alternatively, any or all of the substances may be pre-filled in one or more containers. The substances are preferably introduced into the containers via a set of access ports (e.g., Luer lock access ports) present in a cartridge of the first container management subsystem, but additionally or alternatively, they may be introduced by other suitable methods. The access ports may additionally or alternatively be used to remove the substances from the containers.

[0106] S100 is preferably performed at the beginning of method 200, but additionally or alternatively, it can be performed at other points in method 200 and / or at any other suitable point.

[0107] Method 200 may optionally include a step S200 in which a set of containers is received in a first container management subsystem, which serves to prepare the containers for processing in a subsequent process of Method 200. S200 can be performed before S100, after S100, without performing S100, and / or at any other point in time. The containers are preferably received in a cartridge (e.g., a canister) of the first container management subsystem and placed therein. The cartridge is placed in a receiving opening of the automated equipment, but additionally or alternatively, it may be appropriately received in the first container management subsystem.

[0108] Method 200 preferably includes step S300 in a first container management subsystem which operates a set of containers and / or processes a set of materials, which has the function of initializing, preparing and / or performing at least a portion of the separation process using buoyancy particles. S300 is preferably performed while the chambers of the containers are fluidically separated (e.g., as described above), but some or all of S300 may also be performed while two or more chambers are fluidly coupled. S300 preferably includes rotating the containers (e.g., via the rotation of the cartridge) to facilitate mixing of the materials (e.g., end-over-end mixing, spin, etc.). The mixing of the materials is configured to facilitate the bonding of buoyancy particles with their target material (e.g., increasing the exposure of buoyancy particles to the target material), but additionally or alternatively, it may include moving the containers (e.g., by the translation of the cartridge) and / or any other process.

[0109] Method 200 may optionally include step S400, in which a set of containers is received in a second container management subsystem, which serves to initiate additional processing of the material (e.g., S500). S400 is preferably performed by moving (e.g., translating) the cartridge containing the containers from the receiving opening of the first container management subsystem to the mixing subsystem (e.g., bucket mixer) of the second container management subsystem, but may optionally include any other processing. S400 is preferably performed after S300, but may optionally be performed at any other appropriate time.

[0110] Method 200 preferably includes step S500 of manipulating a set of containers and / or processing a set of materials in a second container management subsystem, which serves to separate buoyancy particles and bound materials from the rest of the set of materials. S500 is preferably performed in response to S400, but can be performed additionally or alternatively at any other appropriate time.

[0111] S500 may include any or all of the following processes: spinning a set of containers; opening and closing valves associated with a set of containers; separating the chambers from each other (e.g., permanently separating them) (e.g., by permanently closing the valves, by permanently closing the valves, by permanently closing the containers after separation, etc.); and / or other processes.

[0112] Alternatively, multiple processes (e.g., multiple mixing processes, end-over-end mixing and spinning, mixing and centrifugation, etc.) can be performed at a single location on an automated device and / or in a single apparatus (hereinafter also equivalently referred to as a single mixing apparatus), the single apparatus may be configured to include multiple rotation axes, translation mechanisms, multiple actuators and / or types of actuators, and / or any other one or more components.

[0113] Method 200 may optionally include step S600 of removing a set of containers and / or a set of substances from an automated device, which serves to enable the use and / or further processing of the substances. In one example, S600 may include adding substances to a consumable (e.g., without user intervention, with user intervention, etc.), discharging substances from a consumable (e.g., without user intervention, with user intervention, etc.), and / or any other optional process. S600 is preferably performed in response to S500, but additionally or alternatively, it may be performed at any other appropriate time. Some or all of S600 may be performed in an automated way (e.g., without user intervention), a semi-automated way (e.g., partially automated with minimal user intervention, etc.), an unautomated way, and / or in other ways.

[0114] Additionally or alternatively, method 200 may include any other suitable process.

[0115] 5. Embodiments In a first aspect of the system, a system for separating a target substance from a total volume with the assistance of buoyancy comprises a first chamber defined by a rigid first container, a second chamber defined by a rigid second container, a deformable conduit positioned between the rigid first container and the rigid second container and configured to selectively fluidically connect the first and second chambers, and a housing comprising a valve configured to connect to the deformable conduit, wherein the valve is operable in a set of operating modes (such as a first mode in which the first chamber is fluidly disconnected from the second chamber, and a second mode in which the first chamber is fluidly connected to the second chamber). The housing may include an automated device comprising a first processing subsystem (to which the housing can be coupled when the valve is operating in a first mode, and the first processing subsystem is configured to mix the target substance in the total volume with a set of buoyancy particles) and a second processing subsystem (to which the housing can be coupled when the valve is operating in a second mode, and the second processing subsystem is configured to separate the target substance from the total volume using the set of buoyancy particles) and / or any other optional components.

[0116] In a first aspect of the method, the method includes the steps of: receiving and / or retrieving a protocol selection (e.g., human T cell negative selection, human T cell positive selection, etc.) (e.g., from a user via a user interface); triggering the start of the protocol; receiving one or more sets of consumables in a first mixing subsystem of an automated instrument; receiving and / or automatically adding substances (e.g., culture medium, sample, cells, antibody, etc.) to one or more chambers of the consumables (e.g., via a set of ports and / or tubes); starting and performing one or more of the first mixing processes (e.g., end-over-end mixing, modified end-over-end mixing, etc.) over a period of time (e.g., 10-15 minutes); optionally adding one or more additional substances (e.g., microbubbles) to one or more chambers; and optionally modifying the first mixing process. The process may include steps of performing additional portions and / or iterations (e.g., with the same mixing parameters, with different mixing parameters) (e.g., for 10-15 minutes), optionally transferring the container to a second mixing subsystem (e.g., a centrifuge, swing bucket centrifuge, etc.) (e.g., via an actuating element, automatically, via manual user intervention), spinning the container in the second mixing subsystem (e.g., for 2-3 minutes, for a shorter period than mixing in the first mixing subsystem, for a longer period than mixing in the first mixing subsystem, etc.) (e.g., at a slower speed than a conventional centrifuge, and at a slower speed than mixing in the first mixing subsystem), thereby enabling separation (e.g., microbubbles and bound material move to the upper chamber, different compositions between chambers, etc.), optionally draining the material from one or more chambers (e.g., into a bag or other new container), and / or any other optional processes.

[0117] In another aspect of this method, the entire mixing process is carried out in a single mixing subsystem (for example, configured to rotate the material in multiple directions / around multiple different axes).

[0118] In another aspect of this method, the method comprises the steps of: receiving a total volume in a first rigid container, comprising a set of target material and a set of buoyancy particles, wherein the set of buoyancy particles is configured to bind with the set of target material; and receiving a housing in a first processing subsystem of an automated device, wherein the housing further comprises a second rigid container, a deformable conduit positioned between the first rigid container and the second rigid container, and a valve connected to the deformable conduit, wherein the valve selectively allows fluid communication between the first rigid container and the second rigid container via the deformable conduit, and the housing is configured such that the valve is closed. The process may include any or all of the following steps: a step which can only be coupled to a first processing subsystem; a step which involves operating the first processing subsystem, which includes mixing the contents of the first rigid container to facilitate the binding of the target material set to the buoyancy particle set; a step which involves receiving the housing in a second processing subsystem of an automated device, wherein the housing can only be coupled to the second processing subsystem if the valve is open; and a step which involves operating the second processing subsystem, which includes mixing the contents of the first and second rigid containers to facilitate the separation of the target material from the rest of the total volume.

[0119] Additionally or alternatively, the system and / or method may be adequately configured in other ways.

[0120] For the sake of brevity, details have been omitted, but preferred embodiments include any combination and substitution of various system components and various method processes, the method processes can be executed sequentially or simultaneously in any suitable order.

[0121] Embodiments of the systems and / or methods may include any combination and substitution of various system components and various method processes, and one or more instances of the methods and / or processes described herein may be executed asynchronously (e.g., sequentially), simultaneously (e.g., simultaneously, in parallel), or in any other suitable order by one or more instances of the systems, elements and / or entities described herein, and / or using them. The following system and / or method components and / or processes may be used in conjunction with, in addition to, instead of, or in combination with all or part of the systems and / or methods disclosed in the above-mentioned applications. Each of those applications is incorporated herein by reference in its entirety.

[0122] Additional or alternative embodiments implement the methods and / or processing modules described above on a non-temporary computer-readable medium that stores computer-readable instructions. These instructions can be executed by computer-executable components integrated into the computer-readable medium and / or processing system. Computer-readable medium includes any suitable computer-readable medium, such as RAM, ROM, flash memory, EEPROM, optical devices (CD or DVD), hard drives, floppy drives, non-temporary computer-readable medium, or any suitable device. Computer-executable components may include computing systems and / or processing systems connected to the non-temporary computer-readable medium (e.g., one or more processors located in the same location or distributed, remote or local processors), such as a CPU, GPU, TPU, microprocessor or ASIC, but alternatively or additionally, instructions may be executed by any suitable dedicated hardware device.

[0123] Those skilled in the art can modify and change preferred embodiments of the present invention without departing from the scope of the invention as defined in the following claims, as can be seen from the detailed description above, the drawings and the claims.

Claims

1. A system for separating a target substance from the total volume with the assistance of buoyancy, - A first chamber, which is defined by a rigid first container, - A second chamber, the second chamber defined by a rigid second container, - A deformable conduit positioned between the rigid first container and the rigid second container, configured to selectively fluidize the first chamber and the second chamber, ・It is a housing, - Includes a valve configured to connect to the deformable conduit, the valve is - A first mode in which the first chamber is fluidly separated from the second chamber, A housing that is operable in a set of operating modes, including a second mode in which the first chamber is fluidly connected to the second chamber, • It is an automated device, - A first processing subsystem wherein the housing can be coupled to the first processing subsystem when the valve is operating in the first mode, and the first processing subsystem is configured to mix the target substance in the total amount with a set of buoyancy particles, An automated device comprising: a second processing subsystem wherein the housing can be coupled to the second processing subsystem when the valve is operating in the second mode, and the second processing subsystem is configured to separate a target substance from the total amount using the set of buoyancy particles. A system characterized by comprising the following features.

2. In the system described in claim 1, A system characterized in that the housing cannot be physically coupled to the first processing subsystem when the valve is operating in the second mode.

3. In the system described in claim 1, A system characterized in that the transition between sets of operating modes is achieved by the relative rotation of the set of valve components.

4. In the system described in claim 3, A system characterized in that at least one of the valve component sets further defines a set of projections, and the relative rotation further adjusts the position of the projections relative to the outer surface of the housing.

5. In the system described in claim 4, A system characterized in that the housing cannot be physically coupled to the second processing subsystem when the valve is operating in the first mode based on the position of the projection.

6. In the system described in claim 1, The system is characterized in that the mixing of the target substance and the set of buoyancy particles in the total amount is performed according to a mixing protocol executed by the first processing subsystem, wherein the protocol specifies the rotation speed.

7. In the system described in claim 6, A system characterized in that at least one of the rotational speed and the rotational angle range is determined at least partially based on the total volume.

8. In the system described in claim 1, The system is characterized in that the valve is located on the outside of the outer wall of the deformable conduit.

9. In the system described in claim 8, The system is characterized in that the valve comprises a set of a plurality of valve components that are rotatable relative to each other, the transition from the second mode to the first mode includes reducing the diameter of an opening defined by the set of valve components by rotation, and the deformable conduit is located within the opening.

10. In the system described in claim 1, A system characterized in that the target substance includes cells, and the total amount includes the amount of blood.

11. A method for automatically assisting the buoyant separation of a set of target substances from a total amount, In the first rigid container, - The total amount including the set of target substances, - A step of receiving a set of buoyancy particles configured to bind with the set of target material, In the first processing subsystem of the automated equipment, - A step of receiving a housing which includes the first rigid container, wherein the housing further - A second hard container, - A deformable conduit positioned between the first rigid container and the second rigid container, - A valve connected to the deformable conduit, which selectively enables fluid communication between the first rigid container and the second rigid container via the deformable conduit, - The housing is connectable to the first processing subsystem only when the valve is in a closed configuration. - A step of operating the first processing subsystem, comprising mixing the contents of the first rigid container to promote the binding of the target material set and the buoyancy particle set, - A step in which the housing is received in the second processing subsystem of the automated equipment, wherein the housing can be coupled to the second processing subsystem only when the valve is in an open configuration. A method characterized by comprising the step of operating the second processing subsystem, which involves mixing the contents of the first and second rigid containers to facilitate the separation of the target substance from the rest of the total amount.

12. In the method according to claim 11, A method characterized in that the first and second processing subsystems are separate and distinct.

13. In the method according to claim 11, A method characterized in that a first protocol defines a first set of parameters, the first set of parameters includes a rotational speed around a first axis, a second protocol defines a second set of parameters, the second set of parameters includes a rotational speed around a second axis, and the first axis is not parallel to the second axis.

14. In the method according to claim 13, A method characterized in that the first processing subsystem is configured for end-over-end mixing around the first axis, and the second processing subsystem is configured for centrifugal separation around the second axis.

15. In the method according to claim 14, The method is characterized in that the centrifugal separation includes swing bucket centrifugal separation.

16. In the method according to claim 11, A method characterized in that a first protocol defines a first set of parameters, the first set of parameters including a rotational speed and a rotational angle range, and the first set of parameters is determined at least in part based on the volume of the total amount.

17. In the method according to claim 11, A method characterized in that the valve is located on the outside of the outer wall of the deformable conduit.

18. In the method according to claim 17, The method is characterized in that the valve comprises a set of a plurality of valve components that are rotatable relative to each other, the transition from the second mode to the first mode includes reducing the diameter of an opening defined by the set of valve components by rotation, and the deformable conduit is located within the opening.

19. In the method according to claim 17, A method characterized in that achieving the closed configuration includes preventing the flow of fluid between the first and second rigid containers across the deformable conduit.

20. In the method according to claim 17, The method is characterized in that the open configuration allows for fluid flow between the first and second rigid containers.

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