Combination fluidic switch for use in automated cell engineering systems - Patents.com

Abstract

The present disclosure provides combination fluidic switches that allow selection of a single fluid flow path while controlling multiple flow paths. The combination fluidic switches can be used in a variety of biological systems and processes and can be included in automated cell engineering systems.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 173,719, entitled “COMBINATORIAL FLUID SWITCH FOR USE IN AUTOMATED CELL ENGINEERING SYSTEMS,” filed April 12, 2021, the entire contents of which are incorporated herein by reference.

[0002] The present disclosure provides combination fluidic switches that allow selection of a single fluid flow path while controlling flow through multiple flow paths. The combination fluidic switches can be used in a variety of biological systems and processes and can be included in automated cell engineering systems. [Background technology]

[0003] As the clinical adoption of advanced cell therapies is expected to accelerate, attention is turning to fundamental manufacturing strategies to ensure these therapies benefit patients worldwide. Although cell therapies hold great clinical potential, high manufacturing costs relative to reimbursement present high barriers to commercialization. Thus, the need for cost-effectiveness, process efficiency, and product consistency is driving automation efforts in many cell therapy segments.

[0004] The generation of therapeutic cell populations will require the automation of various processes, including the integration of cell activation, transduction, and expansion into commercial manufacturing platforms for the translation of these important therapies to broad patient populations.

[0005] In such automated cell engineering systems, it is often necessary to control one or more flows through various valve constructs to deliver cells, media, vector solutions, etc. to desired areas or chambers of the system. The present invention provides a combination fluidic switch design for controlling this fluid flow using a reduced number of valves. Summary of the Invention

[0006] In some embodiments, provided herein is a combination fluid switch for controlling fluid flow through a plurality of fluid flow paths, the combination fluid switch comprising: a plurality of fluid inputs; a first two-position valve having individual fluid flow paths through which fluid from the plurality of fluid inputs is directed; a second two-position valve having individual fluid flow paths through which fluid from the plurality of fluid inputs is directed; and a plurality of fluid outputs, the combination fluid switch being configured to control the flow of fluid from the first fluid input when the first two-position valve is in an open position and when the second two-position valve is in an open position. a combination fluid switch configured to allow fluid flow from the first two-position valve to the first fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, a fluid flow from the second fluid input to the second fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, a fluid flow from the third fluid input to the third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and a fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0007] In further embodiments, there is provided herein a combination fluid switch for controlling fluid flow through at least four fluid flow paths, comprising: first, second, third, and fourth fluid inputs; a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; and first, second, third, and fourth fluid outputs, the combination fluid switch being configured to operate when the first two-position valve is in an open position and the second two-position valve is in a closed position. a combination fluid switch configured to allow fluid flow from a first fluid input to a first fluid output when the first two-position valve is in an open position, a fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, a fluid flow from a third fluid input to a third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and a fluid flow from a fourth fluid input to a fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0008] In still further embodiments, provided herein is a system for controlling fluid flow through at least sixteen fluid flow paths, comprising: a first combination fluid switch, the first combination fluid switch comprising: first, second, third, and fourth fluid inputs; a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; and first, second, third, and fourth fluid outputs, the first combination fluid switch comprising: the valve is configured to allow fluid flow from the first fluid input to the first fluid output when the first two-position valve is in an open position and the second two-position valve is in an open position, fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, fluid flow from the third fluid input to the third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position. a first combination fluid switch; and a second combination fluid switch fluidically connected to the first combination fluid switch, the second combination fluid switch comprising: a fifth, sixth, seventh, and eighth fluid input; a third two-position valve having four fluid flow paths through which fluid from the fifth, sixth, seventh, and eighth fluid inputs is directed; a fourth two-position valve having four fluid flow paths through which fluid from the fifth, sixth, seventh, and eighth fluid inputs is directed; and a fifth, sixth, seventh, and eighth fluid output, the second combination fluid switch being configured such that the third two-position valve is in an open position and the fourth two-position valve is in a closed position. a second combination fluid switch configured to allow fluid flow from the fifth fluid input to the fifth fluid output when the two-position valve is in the open position, fluid flow from the sixth fluid input to the sixth fluid output when the third two-position valve is in the closed position and the third two-position valve is in the open position, fluid flow from the seventh fluid input to the seventh fluid output when the third two-position valve is in the open position and the fourth two-position valve is in the closed position, and fluid flow from the eighth fluid input to the eighth fluid output when the third two-position valve is in the closed position and the fourth two-position valve is in the closed position.The system is provided.

[0009] In yet another embodiment, provided herein is a system for controlling fluid flow through at least 16 fluid flow paths, comprising a plurality of combination fluid switches, each combination fluid switch comprising first, second, third, and fourth fluid inputs, a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, and a first, second, third, and fourth fluid output, A system is provided in which the combination fluid switch is configured to allow fluid flow from a first fluid input to a first fluid output when the first two-position valve is in an open position and the second two-position valve is in an open position, fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, fluid flow from the third fluid input to the third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0010] In still further embodiments, there is provided herein an automated biological production system comprising: a sealable housing; and a cassette contained within the sealable housing, the cassette comprising a cell culture chamber; a combination fluidic switch as described herein; a pumping system fluidly connected to the cell culture chamber and the combination switch; one or more of a temperature sensor, a pH sensor, a glucose sensor, a lactose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor; and a mechanism for automatically regulating one or more of a temperature, a pH level, a glucose level, a lactose level, an oxygen level, a carbon dioxide level, and an optical density.

[0011] In an additional embodiment, provided herein is an automated biological production system comprising: a sealable housing; and a cassette contained within the sealable housing, the cassette comprising a cell culture chamber, a system as described herein, a pumping system fluidly connected to the cell culture chamber and the system, one or more of a temperature sensor, a pH sensor, a glucose sensor, a lactose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor, and a mechanism for automatically regulating one or more of a temperature, a pH level, a glucose level, a lactose level, an oxygen level, a carbon dioxide level, and an optical density.

[0012] Also provided herein is a combination fluid switch comprising a housing having two opposing sides, each side having four openings passing therethrough, and two two-position valves disposed within the housing, each valve having four openings passing therethrough, the side openings and the two-position valve openings configured to receive tubes therethrough to create four flow paths within the combination fluid switch, and the two-position valves are movable within the housing to allow fluid flow through only one flow path at a time.

[0013] In a further embodiment, there is provided herein a combination fluid switch comprising: a support base comprising a plurality of compartments extending above the support base, the two raised portions comprising two recessed portions, the two raised portions configured to allow tubing to pass therethrough to create four fluid flow paths, the two recessed portions each comprising four stationary compression members extending above the support base; and two two-position valves having openings to allow the stationary compression members to pass therethrough, the valves further comprising four movable compression members configured to compress against complementary stationary compression members on the support base thereby contracting tubing between the movable compression members and the complementary stationary compression members, the two-position valves configured to slide along the support base and allow fluid flow through only one flow path at a time.

[0014] In still a further embodiment, there is provided herein a combination fluid switch comprising: a first two-position valve having at least three stationary compression members, two movable compression members integrated into the valve, and a second two-position valve having three movable compression members integrated into the valve, wherein the movable compression members of the first and second two-position valves are configured to compress against complementary stationary compression members, thereby constricting a tube between the movable compression members and the complementary stationary compression members, and the two-position valve is configured to slide and allow fluid flow through one flow path at a time.

[0015] In an additional embodiment, there is provided herein a combination fluid switch for controlling fluid flow through a plurality of fluid flow paths, comprising a plurality of fluid inputs, a first control valve through which fluid from the plurality of fluid inputs is directed, a second control valve having individual fluid flow paths through which fluid from the plurality of fluid inputs is directed, and a plurality of fluid outputs, wherein the combination fluid switch is configured to allow fluid flow through designated combinations of the fluid inputs and fluid outputs.

[0016] In a further embodiment, there is provided herein a method of controlling fluid flow in a closed cell engineering system comprising providing a plurality of fluid inputs, providing a plurality of fluid outputs, providing a plurality of flow paths connecting the fluid inputs to the fluid outputs, and providing a plurality of valves to control flow in the flow paths, thereby controlling the direction, rate, duration and / or interval of fluid flow in the flow paths. [Brief description of the drawings]

[0017] [Figure 1A] 1 illustrates a combination fluidic switch according to an embodiment herein. [Figure 1B] 1 illustrates a combination fluidic switch according to an embodiment herein. [Figure 1C] 1 illustrates a combination fluidic switch according to an embodiment herein. [Figure 1D] 1 illustrates a combination fluidic switch according to an embodiment herein. [Figure 2A] 1 illustrates an exemplary combination fluidic switch. [Figure 2B] 1 illustrates an exemplary combination fluidic switch. [Figure 2C] 1 illustrates an exemplary combination fluidic switch. [Figure 2D] 1 illustrates an exemplary combination fluidic switch. [Figure 3A] 1 illustrates a system for controlling fluid flow according to an embodiment herein. [Figure 3B] 1 illustrates a system for controlling fluid flow according to an embodiment herein. [Figure 4A] 1 shows a representation of a cassette including a combination fluidic switch according to an embodiment herein. [Figure 4B] 1 illustrates an automated biological production system, according to an embodiment of the present disclosure. [Figure 4C] 1 illustrates an automated biological production system, according to an embodiment of the present disclosure. [Diagram 5]1 illustrates an example of a combination fluidic switch according to embodiments herein. [Figure 6] 1 illustrates a further example of a combination fluidic switch according to embodiments herein. [Figure 7A] 13 illustrates additional examples of combination fluidic switches according to embodiments herein. [Figure 7B] 13 illustrates additional examples of combination fluidic switches according to embodiments herein. [Figure 7C] 13 illustrates additional examples of combination fluidic switches according to embodiments herein. [Figure 7D] 13 illustrates additional examples of combination fluidic switches according to embodiments herein. [Figure 8A] 1 illustrates the integration of a combination fluidic switch into an automated cell engineering system, according to embodiments herein. [Figure 8B] 1 illustrates the integration of a combination fluidic switch into an automated cell engineering system, according to embodiments herein. [Figure 8C] 1 illustrates the integration of a combination fluidic switch into an automated cell engineering system, according to embodiments herein. [Figure 9] 1 illustrates the connections of the system described herein. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] It will be understood that the specific embodiments shown and described herein are examples and are not intended to otherwise limit the scope of the application in any way.

[0019] Published patents, patent applications, websites, company names, and scientific literature mentioned in this specification are incorporated herein by reference in their entirety to the same extent as if each was specifically and individually indicated to be incorporated by reference. Any conflict between any reference cited herein and the specific teachings of this specification shall be resolved in favor of the latter. Similarly, any conflict between the art definition of a word or phrase specifically taught in this specification and the definition of the word or phrase shall be resolved in favor of the latter.

[0020] As used herein, the singular forms "a", "an", and "the" specifically include the plurals of the terms they refer to, unless the content clearly indicates otherwise. The term "about" is used herein to mean approximately, within the region of, approximately, or around. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the stated numerical values. In general, the term "about" is used herein to modify a numerical value above and below the stated value with a variance of 20%.

[0021] Unless otherwise defined, technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this application pertains. Reference is made herein to various methodologies and materials known to those skilled in the art.

[0022] In an embodiment, a combination fluid switch is provided herein for controlling fluid flow through multiple fluid flow paths. As used herein, a "fluid switch" refers to a combination of at least one valve and at least one fluid flow path, where the valve controls the fluid flow through the flow path, and in an embodiment, either allows the fluid flow ("on") or stops the fluid flow completely ("off"). A "combinatorial fluid switch" refers to a combination of multiple valves that control the fluid flow through multiple fluid flow paths in various combinations of open and closed positions. The concept of a "combinatorial" switch comes from the fact that multiple valves can be arranged together to allow the control of multiple flow paths.

[0023] 1A-1D show an exemplary combination fluid switch including a first two-position valve having multiple fluid inputs and individual fluid flow paths through which fluid from the multiple fluid inputs is directed, a second two-position valve having individual fluid flow paths through which fluid from the multiple fluid inputs is directed, and multiple fluid outputs. As described herein, the combination fluid switch is configured to allow fluid flow from a first fluid input to a first fluid output when the first two-position valve is in an open position and the second two-position valve is in an open position, fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, fluid flow from the third fluid input to the third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0024] In contrast to the combination fluid switches described herein, which each have two positions, open and closed, and each have one actuator, one valve, and one tube, the theoretical maximum number of combinations that can be achieved with a valve or on / off switch is 2.n where n is the number of valves involved. Thus, for two valves, the number of achievable combinations is four.

[0025] Thus, as shown in Figures 1A-1D, a combination fluid switch 100 can be constructed using two switches that allow control of four fluid flow paths. Thus, in an embodiment, a combination fluid switch is provided herein for controlling fluid flow through at least four fluid flow paths. As used herein, "fluid flow-path" refers to a conduit, channel, tube, tunnel, or similar structure that allows a fluid (preferably a liquid) to pass through a flow path from a flow path inlet to a flow path outlet.

[0026] 1A, the combination fluid switch 100 preferably includes a first fluid input 102, a second fluid input 104, a third fluid input 106, and a fourth fluid input 108. The combination switch also preferably includes a first two-position valve 120 having four fluid flow paths (152, 154, 156, and 158) through which fluid from the first input 102, the second input 104, the third input 106, and the fourth input 108 is directed. The combination switch also preferably includes a second two-position valve 122 through which fluid from the first input 102, the second input 104, the third input 106, and the fourth input 108 is directed. As shown in FIG. 1A, the combination switch 100 also includes a first fluid output 142, a second fluid output 144, a third fluid output 146, and a fourth fluid output 148.

[0027] As exemplarily shown in FIGS. 1A-1D , combination switch 100 is configured to allow (when valves 120 and 122 are open “0” or closed “1”): a fluid flow from the first fluid input 102 to the first fluid output 142 (via flow path 152) when the first two-position valve is in an open position and the second two-position valve is in an open position; a fluid flow from the second fluid input 104 to the second fluid output 144 (via flow path 154) when the first two-position valve is in a closed position and the second two-position valve is in an open position; a fluid flow from the third fluid input 106 to the third fluid output 106 (via flow path 156) when the first two-position valve is in an open position and the second two-position valve is in a closed position; and Fluid flow from the fourth fluid input 108 to the fourth fluid output 148 (via flow path 158) when the first two-position valve is in the closed position and the second two-position valve is in the closed position.

[0028] 1B, when the first two-position valve 120 is closed ("1"), flow path 152 is blocked (see top of FIG. 1B) by the first two-position valve 120, as is flow path 156 (see center of FIG. 1B), while flow path 158 is blocked (see bottom of FIG. 1B) by the second two-position valve 122. However, flow path 154 is open, and thus fluid can flow from input 104 to output 144 via flow path 154.

[0029] 2A-2D show an exemplary combination switch 100, where each of the fluid inputs comprises a tube, and the fluid flow paths (152, 154, 156, 158) also preferably comprise a tube. As shown in FIGS. 2A-2D, in an embodiment, the two-position valves 120 and 122 are trumpet valves, i.e., valves that have two positions (open "0" or closed "1") and can compress or pinch the flow paths (e.g., tubing). Preferably, as shown in FIGS. 2A-2D, in an embodiment, the fluid inputs, fluid flow paths, and fluid outputs comprise at least four tubing lines. In a preferred embodiment, the trumpet valves include a shield or cover (preferably a silicone cover or similar material) that limits contamination and maintains the sterility of the trumpet valves and thus the flow paths in which they are utilized.

[0030] Exemplary materials for use in tubing for the flow paths, inputs, and outputs are known in the art and include, for example, various polymers (e.g., silicone tubing), plastics, glass, metals, ceramics, composites, and the like.

[0031] In additional embodiments, each of the fluid inputs (102, 104, 106, and 108) may be supplied from a single fluid source 110 (see, e.g., FIG. 1A). Preferably, the fluids in the fluid flow paths are not allowed to mix with each other. This may be accomplished by using tubing components for each of the fluid flow paths, or separate conduits or other passages that do not interact with each other.

[0032] In a further embodiment, a system for controlling fluid flow is provided herein. In an exemplary embodiment, the system controls fluid flow through at least 16 fluid flow paths. As shown in FIG. 3A, the system 300 suitably comprises a first combination fluid switch 100 as described herein, e.g., a first, second, third, and fourth fluid input, a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, and a first, second, third, and fourth fluid output. As described herein, the first combination fluid switch is configured to allow fluid flow from the first fluid input to the first fluid output when the first two-position valve is in an open position and the second two-position valve is in an open position, fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, fluid flow from the third fluid input to the third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0033] The system 300 also includes a second combination fluid switch 100' fluidically connected to the first combination fluid switch (see the fluidic connection between the first combination fluid switch 100 and the second combination fluid switch 100' in FIG. 3A). The second combination fluid switch 100' preferably comprises a fifth, sixth, seventh, and eighth fluid input, a third two-position valve having four fluid flow paths through which fluids from the fifth, sixth, seventh, and eighth fluid inputs are directed, a fourth two-position valve having four fluid flow paths through which fluids from the fifth, sixth, seventh, and eighth fluid inputs are directed, and a fifth, sixth, seventh, and eighth fluid output. As described herein, the second combination fluid switch 100' is configured to allow fluid flow from the fifth fluid input to the fifth fluid output when the third two-position valve is in an open position and the fourth two-position valve is in an open position, fluid flow from the sixth fluid input to the sixth fluid output when the third two-position valve is in a closed position and the third two-position valve is in an open position, fluid flow from the seventh fluid input to the seventh fluid output when the third two-position valve is in an open position and the fourth two-position valve is in a closed position, and fluid flow from the eighth fluid input to the eighth fluid output when the third two-position valve is in a closed position and the fourth two-position valve is in a closed position.

[0034] It should be understood that the numbers of fluid inputs, flow paths, and fluid outputs are utilized merely to illustrate the combinatorial nature of the switches described herein, and the numerical numbering does not imply that the valves must be utilized in any particular order or orientation (i.e., the valves may be selected in any order and in any sequence).

[0035] In yet a further embodiment, a system 300 for controlling fluid flow through at least 16 fluid flow paths includes a plurality of combination fluid switches (e.g., 100 and 100′), each combination fluid switch comprising first, second, third, and fourth fluid inputs, a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, and a first, second, third, and fourth fluid output. the first combination fluid switch is configured to allow fluid flow from the first fluid input to the first fluid output when the first two-position valve is in the open position and the second two-position valve is in the open position, fluid flow from the second fluid input to the second fluid output when the first two-position valve is in the closed position and the second two-position valve is in the open position, fluid flow from the third fluid input to the third fluid output when the first two-position valve is in the open position and the second two-position valve is in the closed position, and fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in the closed position and the second two-position valve is in the closed position.

[0036] In a preferred embodiment, each of the combination fluidic switches (e.g., 100 and 100') are fluidly connected to one another. As used herein, "fluidly connected" refers to a junction or encounter between two fluidic switches that allows fluids to pass from one switch to the other without loss of volume and without mixing between the fluids (unless mixing is desired).

[0037] In embodiments, system 300 may include four or more combination fluidic switches, such as 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, or 20 or more fluidic switches.

[0038] As described throughout, preferably the fluid inputs, fluid flow paths, and fluid outputs include tubing and the valves are trumpet valves. In an exemplary embodiment, the fluid inputs, fluid flow paths, and fluid outputs consist of at least eight tubing lines.

[0039] Figure 3B shows the 16 different flow paths that can be achieved through the use of system 300 when there are four inputs, four flow paths, and four outputs per valve pair. As shown in Figure 3B, each of the two position valves (four in total) has an open position (0) and a closed position (1). By selecting the two available positions of each of the four valves in all of the possible combinations (16 in total), the 16 different flow paths are selected. The matrix at the bottom right of Figure 3B shows the 16 valve position combinations and the flow path matrix shows the 16 different flow paths.

[0040] As shown, using just four two-position valves allows for a total of 16 flow paths (2 4 ), which illustrates a significant advantage of system 300, which includes fluidly connected combination switches 100, dramatically expanding the number of flow paths that can be achieved compared to conventionally oriented valves.

[0041] In yet a further embodiment, provided herein is an automated biological production system that includes a combination fluidic switch as described herein. Figure 4A shows a schematic diagram of an automated biological production system 400 as described herein, and Figures 4B and 4C show examples of automated biological production systems 400 that include cassettes for the production of biological products including antibodies, proteins, cells, etc.

[0042] As described herein, the cassettes and methods are utilized and performed in a fully enclosed automated biological production system 400 (see Figures 4B and 4C), e.g., an automated cell engineering system, suitable for performing steps such as activation, transduction, expansion, enrichment, and harvesting. For example, a cell engineering system for the automated production of genetically modified immune cells, including CAR T cells, is described in U.S. Published Patent Application No. 2019 / 0169572, the disclosure of which is incorporated herein by reference in its entirety, and is also referred to herein as the automated cell engineering system, COCOON™, or COCOON™ System.

[0043] For example, a user can provide an automated cell engineering system pre-filled with cell cultures and reagents (e.g., activation reagents, vectors, cell culture media, nutrients, selection reagents, etc.), as well as parameters for cell production (e.g., starting number of cells, type of media, type of activation reagent, type of vector, number or dosage of cells produced, etc.). The automated cell engineering system can perform various automated methods, including methods for producing genetically modified immune cell cultures, including CAR T cells, without further input from the user. In some embodiments, a fully enclosed automated cell engineering system minimizes contamination of the cell cultures by reducing exposure of the cell cultures to non-sterile environments. In additional embodiments, a fully enclosed automated cell engineering system minimizes contamination of the cell cultures by reducing the user's handling of the cells.

[0044] In an embodiment, an automated biological production system (cell engineering system) 400 (see FIGS. 4A-4C) includes a sealable housing 408, a cassette 410 contained within the sealable housing 408, the cassette comprising a cell culture chamber 402, a combination fluidic switch 100 as described herein, and a pumping system 404 fluidly connected to the cell culture chamber 402 and the combination switch 100. The automated biological production system 400 further includes one or more of a temperature sensor, a pH sensor (e.g., 406), a glucose sensor, a lactose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor, as well as mechanisms for automatically adjusting one or more of the temperature, pH level, glucose level, lactose level, oxygen level, carbon dioxide level, and optical density. As shown in FIG. 4A, the combination switch can be positioned within the flow path of the automated biological production system 400 at various locations (see 100 and 100'''').

[0045] As used herein, "enclosable housing" refers to a structure that can be opened and closed, such that the cassette 410 described herein can accommodate and integrate various components such as fluid supply lines, gas supply lines, power sources, cooling connections, heating connections, etc. As shown in Figures 4B and 4C, the enclosable housing can be opened to allow insertion of the cassette (Figure 4C) and closed (Figure 4B) to maintain a closed, sealed environment to enable the cassette to be utilized for various automated processes described herein.

[0046] In other embodiments, an automated biological production system 400 can include one or more of systems 300 and 300', as shown in Figure 4A. In such embodiments, the automated biological production system 400 suitably includes a sealable housing 408, a cassette 410 contained within the sealable housing, the cassette comprising a cell culture chamber 401, a system 300 as described herein, a pumping system 404 fluidly connected to the cell culture chamber and system 300, one or more of a temperature sensor, a pH sensor (e.g., 406), a glucose sensor, a lactose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor, and mechanisms for automatically regulating one or more of the temperature, pH level, glucose level, lactose level, oxygen level, carbon dioxide level, and optical density.

[0047] In an exemplary embodiment, the automated biological production system 400 can further include one or more of a magnetic cell separation device or an electroporation device. The automated biological manufacturing system 400 suitably includes at least 16 fluid flow paths, including, for example, at least 17 fluid flow paths, at least 18 fluid flow paths, at least 19 fluid flow paths, at least 20 fluid flow paths, at least 21 fluid flow paths, at least 22 fluid flow paths, at least 23 fluid flow paths, at least 24 fluid flow paths, at least 25 fluid flow paths, at least 26 fluid flow paths, at least 27 fluid flow paths, at least 28 fluid flow paths, at least 29 fluid flow paths, or at least 30 fluid flow paths.

[0048] As described throughout, the automated biological production system 400 described herein is suitably configured to produce cells, e.g., CAR-T cells.

[0049] As described herein, an automated biological production system 400, including an automated cell engineering system, suitably includes a cassette 410. Thus, in embodiments, provided herein is a cassette for use in an automated cell engineering system including one or more combination fluidic switches 100, and / or a system 300 including a combination fluidic switch 100, as described throughout. As used herein, "cassette" refers to a largely self-contained, removable, and replaceable element of an automated biological production (cell engineering) system, the system including one or more chambers for carrying out various elements of the methods described herein, and suitably including one or more of cell culture media, activation reagents, wash media, etc.

[0050] Cassette 410 preferably includes a cell sample input, which may be a vial or chamber into which a cell sample may be placed prior to introduction or loading into cassette 410. In other embodiments, the cell sample input may simply be a sterile locking tube (e.g., a Luer lock tubing connection, etc.) to which a syringe or cell-containing bag, such as a blood bag, may be connected.

[0051] Exemplary fluid connections that may be used in cassette 410 to connect with the various components, including combination fluidic switch 100 and system 300, include various tubes, channels, and connections known in the art, such as silicone or rubber tubing, luer lock connections, and the like. It should be understood that the fluidly connected components may also include additional elements between each of the components while still maintaining a fluid connection. That is, the fluidly connected components may, but need not, include additional elements such that fluid passing between the components can also pass through these additional elements.

[0052] The pumping system 404 is preferably a peristaltic pump system, although other pumping systems may be utilized. A peristaltic pump refers to a type of positive displacement pump for pumping fluids. The fluid is preferably contained within a flexible tube mounted inside a pump casing, which is often circular. A rotor, with a number of "rollers," "shoes," "wipers," or "lobes" attached to its periphery, compresses the flexible tube. As the rotor rotates, it pinches closed (i.e., "occludes") a portion of the tube under compression, thereby pumping and moving the fluid through the tube. Furthermore, as the tube opens ("rebounds" or "bounces") after passing a cam, fluid flow is induced in the pump. This process is called peristalsis and is used to move fluids through flexible tubes. Typically, there are two or more rollers, or wipers, that occlude the tube and capture a body of fluid between them. The body of fluid is then transported toward the pump outlet.

[0053] As described herein, magnetic separation processes can be utilized to eliminate and separate undesirable cells and debris from a cell population. In such embodiments, magnetic beads or other structures to which biomolecules (e.g., antibodies, antibody fragments, etc.) are attached can interact with the target cells. A variety of magnetic separation methods can then be used to separate the target cell population from undesirable cells, debris, etc. that may be in the cell sample, including the use of filters, columns, flow tubes, or channels with magnetic fields. For example, the target cell population can flow through a tube or other structure and be exposed to a magnetic field. The target cell population can thereby be maintained or retained by the magnetic field, and the undesirable cells and debris can pass through the tube. The magnetic field can then be turned off, allowing the target cell population to pass through additional retention chambers or other areas of the cassette for further automated processing. Additional filtration can include the use of traditional column filtration, or other filtration membranes and structures.

[0054] In an exemplary embodiment, the cell culture chamber 402 is a flat, non-flexible chamber (i.e., made from a substantially non-flexible material such as plastic) that does not easily bend or curve. The use of a non-flexible chamber allows the cells to remain substantially undisturbed. In an embodiment, the overall thickness (i.e., chamber height) of the cell culture chamber 402 is low, on the order of about 0.5 cm to about 5 cm. Preferably, the cell culture chamber has a volume of about 0.50 ml to about 500 ml, or about 1 ml to about 300 ml, more preferably about 50 ml to about 200 ml, or the cell culture chamber has a volume of about 180 ml. The use of a low chamber height (less than 5 cm, preferably less than 4 cm, less than 3 cm, or less than 2 cm) allows for effective medium and gas exchange close to the cells. The ports are configured to allow mixing via fluid recirculation without disturbing the cells. Static vessels of greater height can create concentration gradients that limit oxygen and fresh nutrients in areas close to the cells. By controlling flow dynamics, medium exchange can be performed without disturbing the cells. Medium can be removed from the additional chamber (where no cells are present) without risk of cell loss.

[0055] As described herein, in exemplary embodiments, the cassette is pre-filled with one or more of the following: cell culture, culture medium, cell wash medium, if desired, activation reagents, and / or vectors containing any combination thereof. In further embodiments, these various elements may be added later via suitable injection ports, etc.

[0056] As described herein, in embodiments, the cassette suitably further includes one or more of a pH sensor 406, a glucose sensor (not shown), an oxygen sensor, a carbon dioxide sensor (not shown), a lactate sensor / monitor (not shown), and / or an optical density sensor (not shown). The cassette may also include one or more sampling and / or injection ports. Examples of such sampling and injection ports include access ports for connecting the cartridge to external devices such as an electroporation unit or an additional medium source.

[0057] In an embodiment, the cassette 410 preferably includes a low temperature chamber, which may include a refrigerated area suitable for storage of cell culture media, and a high temperature chamber suitable for activating, transducing, and / or expanding the cell culture. Preferably, the high temperature chamber is separated from the low temperature chamber by a thermal barrier. As used herein, "low temperature chamber" refers to a chamber that is preferably maintained below room temperature, more preferably at about 4°C to about 8°C, to maintain cell media and the like at refrigerated temperatures. As used herein, "high temperature chamber" refers to a chamber that is preferably maintained above room temperature and at a temperature more suitable for enabling cell proliferation and growth, i.e., about 35-40°C, and more preferably at about 37°C. In an embodiment, the high temperature chamber preferably includes the cell culture chamber 206 (also referred to throughout as the proliferation chamber or cell proliferation chamber).

[0058] The fluid pathways, including the gas exchange lines, may be made from a gas permeable material, such as silicone. In some embodiments, the automated cell engineering system recirculates oxygen throughout the substantially non-yielding chamber during the cell production method. Thus, in some embodiments, the oxygen level of the cell culture in the automated cell engineering system is higher than the oxygen level of the cell culture in the flexible gas permeable bag. Higher oxygen levels can be important during the cell culture extension step, as increased oxygen levels can support increased cell growth and proliferation.

[0059] In embodiments, the methods and cartridges described herein are utilized in the COCOON® platform (Octane Biotech, Kingston, Ontario), which integrates multiple unit operations on a single turnkey platform. Multiple cell protocols are available for very specific cell processing objectives. To provide efficient and effective automation translation, the methods described utilize the concept of application-specific / sponsor-specific disposable cassettes that combine multiple unit operations, all focused on the core requirements of the final cell therapy product. Multiple automated cell engineering systems 400 can be integrated together into large-scale multi-unit operations to generate large volumes of cells or multiple different cell samples for individual patients.

[0060] The automated cell engineering system also includes a user interface 420 for receiving input from a user. The user interface 420 may be a touch pad, tablet, keyboard, computer terminal, or other suitable interface, allowing a user to input desired controls and criteria to the automated cell engineering system for controlling the automated process and flow paths. Preferably, the user interface is coupled to a computer control system to provide instructions to the automated cell engineering system and control the overall activity of the automated cell engineering system. Such instructions may include when to open or close various valves, when to provide media or cell masses, when to increase or decrease temperatures, etc.

[0061] Automation of unit operations in cell therapy production offers the opportunity for universal benefits across allogeneic and autologous cell therapy applications. In the unique scenario of patient-specific autologous cell products, the benefits of automation are further highlighted by the clinical success of these therapies, but are particularly attractive due to the complexities of small batch GMP compliance, economics, patient traceability, and the critical early detection of process deviations in microlots. With the emergence of complex manufacturing protocols, the value of end-to-end integration of automated unit operations in microlot cell production has been highlighted, a fact that has not been well explored to date. However, with the expected demand for these soon-to-be approved therapies, the implementation of a fully closed end-to-end system can provide a necessary solution to manufacturing bottlenecks such as hands-on time and footprint.

[0062] FIG. 4A illustrates an exemplary setup utilizing two combination fluidic switches (100 and 100''''') and two systems (300 and 300'), each of which includes two combination fluidic switches. As shown, each fluidic switch has the capacity to accept up to four inputs, including one or more flow-through lines, and four outputs. As shown, the inputs can include various connections to the cell culture chamber 402 inputs and outputs, various sampling port inputs and outputs, various media chamber inputs and outputs, various filter inputs and outputs, including magnetic filters, and the like, and pump connections.

[0063] In further embodiments, there is provided herein a combination fluidic switch for controlling fluid flow through multiple fluid flow paths, the combination fluidic switch comprising multiple fluid inputs, a first control valve through which fluids from the multiple fluid inputs are directed, a second control valve having individual fluid flow paths through which fluids from the multiple fluid inputs are directed, and multiple fluid outputs, the combination fluidic switch configured to allow fluid flow through designated combinations of fluid inputs and fluid outputs. A variety of control valves may be utilized in the combination switch, including those described herein and otherwise known in the art, to allow control of flow through multiple inputs, multiple flow paths, and from multiple outputs. The combinational nature of the fluidic switches described herein allows for coordinated control of fluid flow, preferably within an automated biological production system 400, preferably within a cassette 410 of a cell engineering system. The combinational nature of the fluidic switches also allows for control of fluid flow within a closed system, maintaining sterility and hermeticity of the fluid flow system.

[0064] The fluid flow paths described herein suitably connect various temperature zones and can connect various elements of the cell engineering system, including cell separation, cell washing, cell isolation, etc. The flow paths suitably direct various reagents and include cell or virus transport flow paths. The flow paths can also be utilized to direct flow from the cell engineering system (e.g., from the cassette) to external devices such as electroporation or mechanoporation hardware, and microscopes or optics (e.g., cameras), cell counting or cell sorting equipment, etc.

[0065] The scale of the flow paths can also be tailored by using various tube diameters, including mm to mm scales, and by using tubes to create turbulence or increase shear, or to eliminate or reduce turbulence or shear. The flow paths can also allow for the mixing or combination of reagents, cells, viruses, etc., and maintain their desired temperature and flow characteristics (i.e., steady flow or turbulent flow, as desired).

[0066] 5 illustrates an example combination fluidic switch 100 as described herein that includes a housing 502 having two opposing sides 504, each side having four openings 506 passing therethrough, and two two-position valves 508 disposed within the housing, each valve having four openings 506 passing therethrough. As described herein, the openings 506 in the sides 504 and the openings 506 in the two-position valves 508 are configured to receive tubes therethrough to create four flow paths within the combination fluidic switch, and the two-position valves are movable within the housing to allow fluid flow through only one flow path at a time.

[0067] In an exemplary embodiment, the two-position valve is slidable within the housing. In a further embodiment, the two-position valve is rotatable within the housing.

[0068] In a preferred embodiment, the switch 100 shown in FIG. 5 further includes four tubing lines passing through four openings in the side and four openings in the two-position valve.

[0069] In a further embodiment, Figure 6 illustrates a combination fluidic switch 100 comprising a support base 602 comprising two raised portions 604 and two recessed portions 608, the two raised portions 604 comprising a plurality of compartments 606 extending above the support base configured to allow tubing to pass therethrough to create four fluid flow paths. The two recessed portions 608 each comprise four stationary compression members 610 extending above the support base. The switch 100 also includes two two-position valves 612 having openings 614 allowing the stationary compression members 610 to pass therethrough, the valves further comprising four movable compression members 616 configured to compress against complementary stationary compression members 610 on the support base 602, thereby contracting the tubing between the movable compression members 616 and the complementary stationary compression members 610. Preferably, the two-position valve is configured to slide along the support base and to allow fluid flow through only one flow path at a time.

[0070] 6, the two two-position valves 612 are flat valves that slide within two recessed portions 608 of the support base 602. The combination fluid switch 100 also preferably further includes four tubing lines passing through the compartments, the stationary compression member, and the movable compression member.

[0071] 7A-7D show a further combination fluid switch 100. In an embodiment, the switch of FIGS. 7A-7D includes at least three stationary compression members 702, a first two-position 704 valve having two movable compression members 706 integrated into the valve, and a second two-position valve 704 having three movable compression members 706 integrated into the valve. The movable compression members 706 of the first and second two-position valves 704 are configured to compress against the complementary stationary compression members 702, thereby contracting a tube 708 between the movable compression members and the complementary stationary compression members (see FIGS. 7B-7D). The two-position valve 704 is configured to slide and allow fluid flow through one flow path at a time. In an exemplary embodiment, the stationary compression module 702 is disposed within a housing 710. Preferably, the two-position valve is a trumpet valve. As shown in Figures 7B-7D, the combination fluid switch 100 of Figures 7A-7D can further include four tubing lines 708 passing through the stationary compression member 702 and the movable compression member 706 to provide four fluid flow paths. Note that the movable and stationary compression members shown in Figures 7A-7D can also be reversed and still achieve the same control of the flow paths.

[0072] 8A-8C show the integration of six combination fluidic switches 100 into an automated cell engineering system 400. As shown, each system 300 preferably includes two combination switches 100. The systems are mounted on a support manifold 802 of a cassette 410 (with the remainder of the cassette removed for clarity). As described herein, the switches 100 preferably include two two-position valves (120, 122), such as trumpet valves. The use of six combination fluidic switches allows for the integration of a 6×2 flow channel through the automated cell engineering system 400. 2 (i.e., 24) flow path control. In a further embodiment, a combination switch may be integrated utilizing two combination switches 100 at either end with two systems 300 between them, resulting in 4+16+16+4=40 flow path variations or combinations.

[0073] 9 shows the connections of a system 300 including two combination switches 100 in a cassette 410, as described herein. A tubing connection 900 between the two combination switches 100 is also shown.

[0074] Additional Exemplary Embodiments A first embodiment is a combination fluid switch for controlling fluid flow through a plurality of fluid paths, the combination fluid switch comprising: a plurality of fluid inputs; a first two-position valve having individual fluid paths through which fluid from the plurality of fluid inputs is directed; a second two-position valve having individual fluid paths through which fluid from the plurality of fluid inputs is directed; and a plurality of fluid outputs, the combination fluid switch being configured to control fluid flow from the first fluid input to the first two-position valve when the first two-position valve is in an open position and when the second two-position valve is in an open position. a fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position; a fluid flow from the third fluid input to the third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position; and a fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0075] Embodiment 2 includes the combination fluidic switch of embodiment 1, wherein the fluid input comprises a tube.

[0076] Example 3 includes the combination fluidic switch of Example 1 or 2, wherein the fluid flow path comprises a tube.

[0077] A fourth embodiment includes the combination fluid switch according to any one of the first to third embodiments, in which the two-position valve is a trumpet valve.

[0078] A fifth embodiment includes the combination fluid switch according to any one of the first to fourth embodiments, in which the fluid output portion includes a tube.

[0079] A sixth embodiment includes the combination fluid switch of any one of the first to fifth embodiments, wherein the fluid input, the fluid flow path, and the fluid output are comprised of at least four tube lines.

[0080] A seventh embodiment includes a combination fluidic switch according to any one of the first to sixth embodiments, in which each of the fluid inputs is supplied from a single fluid source.

[0081] An eighth embodiment includes a combination fluidic switch according to any one of the first to seventh embodiments, in which the fluids in the fluid flow paths are not allowed to mix.

[0082] A ninth embodiment is a combination fluid switch for controlling fluid flow through at least four fluid flow paths, the combination fluid switch comprising: first, second, third, and fourth fluid inputs; a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; and first, second, third, and fourth fluid outputs, the combination fluid switch being configured to operate when the first two-position valve is in an open position and the second two-position valve is in a closed position. a combination fluid switch configured to allow fluid flow from a first fluid input to a first fluid output when the first two-position valve is in an open position, a fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, a fluid flow from a third fluid input to a third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and a fluid flow from a fourth fluid input to a fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0083] Example 10 includes the combination fluidic switch of example 9, wherein the fluid input comprises a tube.

[0084] An eleventh embodiment includes the combination fluidic switch of any one of the nine or tenth embodiments, wherein the fluid flow path comprises a tube.

[0085] A twelfth embodiment includes the combination fluid switch according to any one of the ninth to eleventh embodiments, wherein the two-position valve is a trumpet valve.

[0086] A thirteenth embodiment includes the combination fluid switch of any one of the ninth to twelfth embodiments, wherein the fluid output comprises a tube.

[0087] A fourteenth embodiment includes the combination fluid switch according to any one of the ninth to thirteenth embodiments, wherein the fluid input portion, the fluid flow path, and the fluid output portion are made up of four tube lines.

[0088] A fifteenth embodiment includes the combination fluidic switch of any one of the ninth to fourteenth embodiments, wherein each of the fluid inputs is supplied from a single fluid source.

[0089] A sixteenth embodiment is a system for controlling fluid flow through at least sixteen fluid flow paths, comprising: a first combination fluid switch having first, second, third, and fourth fluid inputs; a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed; and a first, second, third, and fourth fluid output, the first combination fluid switch being configured to operate when the first two-position valve is in an open position. a first combination fluid switch configured to allow fluid flow from the first fluid input to the first fluid output when the second two-position valve is in an open position, fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, fluid flow from the third fluid input to the third fluid output when the first two-position valve is in the open position and the second two-position valve is in the closed position, and fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in the closed position and the second two-position valve is in the closed position. a second combination fluid switch fluidically connected to the first combination fluid switch, the second combination fluid switch comprising: a fifth, sixth, seventh, and eighth fluid input; a third two-position valve having four fluid flow paths through which fluid from the fifth, sixth, seventh, and eighth fluid inputs is directed; a fourth two-position valve having four fluid flow paths through which fluid from the fifth, sixth, seventh, and eighth fluid inputs is directed; and a fifth, sixth, seventh, and eighth fluid output, the second combination fluid switch being configured to operate when the third two-position valve is in an open position and when the fourth two-position valve is in an open position. a second combination fluid switch configured to allow fluid flow from the fifth fluid input to the fifth fluid output at a certain time, fluid flow from the sixth fluid input to the sixth fluid output when the third two-position valve is in a closed position and the third two-position valve is in an open position, fluid flow from the seventh fluid input to the seventh fluid output when the third two-position valve is in an open position and the fourth two-position valve is in a closed position, and fluid flow from the eighth fluid input to the eighth fluid output when the third two-position valve is in a closed position and the fourth two-position valve is in a closed position.

[0090] Embodiment 17 is a system for controlling fluid flow through at least 16 fluid flow paths, comprising a plurality of combination fluid switches, each combination fluid switch comprising first, second, third, and fourth fluid inputs, a first two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, a second two-position valve having four fluid flow paths through which fluid from the first, second, third, and fourth fluid inputs is directed, and a first, second, third, and fourth fluid output. The fluid switch is a system configured to allow fluid flow from a first fluid input to a first fluid output when the first two-position valve is in an open position and the second two-position valve is in an open position, fluid flow from the second fluid input to the second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position, fluid flow from the third fluid input to the third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position, and fluid flow from the fourth fluid input to the fourth fluid output when the first two-position valve is in a closed position and the second two-position valve is in a closed position.

[0091] Example 18 includes the system of example 16 or 17, wherein each of the combination fluidic switches are fluidly connected to each other.

[0092] Embodiment 19 includes the system of embodiment 16 or 17, comprising four or more combination fluidic switches.

[0093] Example 20 includes the system of any one of Examples 16-19, wherein the fluid input comprises a tube.

[0094] Example 21 includes the system of any one of Examples 16-20, wherein the fluid flow path comprises a tube.

[0095] Example 22 includes the system of any one of Examples 16-21, wherein each of the valves is a trumpet valve.

[0096] Example 23 includes the system of any one of Examples 16-22, wherein the fluid output comprises a tube.

[0097] Example 24 includes the system of any one of Examples 16-23, wherein the fluid input, fluid flow path, and fluid output consist of at least eight tubing lines.

[0098] Embodiment 25 is an automated biological production system comprising: a sealable housing; and a cassette contained within the sealable housing, the cassette comprising a cell culture chamber; a combination fluidic switch as described in embodiment 1; a pumping system fluidly connected to the cell culture chamber and the combination switch; one or more of a temperature sensor, a pH sensor, a glucose sensor, a lactose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor; and a mechanism for automatically regulating one or more of a temperature, a pH level, a glucose level, a lactose level, an oxygen level, a carbon dioxide level, and an optical density.

[0099] Embodiment 26 is an automated biological production system comprising: a sealable housing; and a cassette contained within the sealable housing, the cassette comprising a cell culture chamber; the system of embodiment 8 or 9; a pumping system fluidly connected to the cell culture chamber and the system; one or more of a temperature sensor, a pH sensor, a glucose sensor, a lactose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor; and a mechanism for automatically regulating one or more of a temperature, a pH level, a glucose level, a lactose level, an oxygen level, a carbon dioxide level, and an optical density.

[0100] Embodiment 27 includes an automated biological production system according to embodiment 25 or 26 configured to produce cells.

[0101] Embodiment 28 includes the automated biological production system of any one of embodiments 25 to 27, further comprising a magnetic cell separation device.

[0102] Embodiment 29 includes the automated biological production system of any one of embodiments 25 to 28, further comprising an electroporation device.

[0103] Embodiment 30 includes the automated biological production system of any one of embodiments 25-29, wherein the automated biological production system includes at least 16 fluid flow paths.

[0104] Embodiment 31 includes the automated biological production system of any one of embodiments 25-30, wherein the automated biological production system includes at least 20 fluid flow paths.

[0105] Embodiment 32 is a combination fluid switch comprising a housing having two opposing sides, each side having four openings passing therethrough, and two two-position valves disposed within the housing, each valve having four openings passing therethrough, wherein the openings on the sides and the openings in the two-position valves are configured to receive tubes therethrough to create four flow paths within the combination fluid switch, and the two-position valves are movable within the housing to allow fluid flow through only one flow path at a time.

[0106] Embodiment 33 includes the combination fluid switch of embodiment 32, wherein the two-position valve is slidable within the housing.

[0107] Embodiment 34 includes the combination fluid switch of embodiment 32, wherein the two-position valve is rotatable within the housing.

[0108] Embodiment 35 includes the combination fluid switch of any one of embodiments 32-34, further comprising four tubing lines passing through the four openings in the side and the four openings in the two-position valve.

[0109] Embodiment 36 is a combination fluid switch comprising: a support base with a plurality of compartments extending above the support base, the two raised portions and two recessed portions, the two raised portions configured to allow a tube to pass therethrough to create four fluid flow paths, the two recessed portions each comprising four stationary compression members extending above the support base; and two two-position valves having openings allowing the stationary compression members to pass therethrough, the valves further comprising four movable compression members configured to compress against complementary stationary compression members on the support base, thereby contracting a tube between the movable compression members and the complementary stationary compression members, the two-position valves configured to slide along the support base and allow fluid flow through only one flow path at a time.

[0110] Embodiment 37 includes the combination fluid switch of embodiment 36, wherein the two two-position valves are flat valves that slide within two recessed portions of the support base.

[0111] Embodiment 38 includes the combination fluid switch of embodiment 36 or 37, further comprising four tubing lines passing through the compartment, the stationary compression member, and the movable compression member.

[0112] Embodiment 39 is a combination fluid switch comprising: a first two-position valve having at least three stationary compression members and two movable compression members integrated into the valve; and a second two-position valve having three movable compression members integrated into the valve, wherein the movable compression members of the first and second two-position valves are configured to compress against complementary stationary compression members, thereby contracting a tube between the movable compression members and the complementary stationary compression members, and the two-position valve is configured to slide and allow fluid flow through one flow path at a time.

[0113] Embodiment 40 includes the combination fluid switch of embodiment 39, wherein the static compression module is disposed within the housing.

[0114] Embodiment 41 includes the combination fluid switch of embodiment 39 or 40, wherein the two-position valve is a trumpet valve.

[0115] Example 42 includes the combination fluid switch of any one of Examples 39-41, further comprising four tubing lines passing through the stationary compression member and the movable compression member to provide four fluid flow paths.

[0116] Embodiment 43 is a combination fluid switch for controlling fluid flow through multiple fluid flow paths, comprising: multiple fluid inputs; a first control valve through which fluid from the multiple fluid inputs is directed; a second control valve having individual fluid flow paths through which fluid from the multiple fluid inputs is directed; and multiple fluid outputs, wherein the combination fluid switch is configured to allow fluid flow through designated combinations of the fluid inputs and fluid outputs.

[0117] Embodiment 44 is a method of controlling fluid flow in a closed cell engineering system comprising providing a plurality of fluid inputs, providing a plurality of fluid outputs, providing a plurality of flow paths connecting the fluid inputs to the fluid outputs, and providing a plurality of valves to control flow in the flow paths, thereby controlling the direction, rate, duration and / or interval of fluid flow in the flow paths.

[0118] It will be readily apparent to those skilled in the relevant art that other suitable modifications and adaptations to the methods and applications described herein can be made without departing from the scope of any of the embodiments.

[0119] Although specific embodiments have been illustrated and described herein, it should be understood that the claims should not be limited to the specific forms or arrangements of parts described and illustrated. Although exemplary embodiments are disclosed herein and specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limitation. Modifications and variations of the embodiments are possible in light of the above teachings. It should be understood, therefore, that the embodiments may be practiced otherwise than as specifically described.

[0120] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

Claims

1. 1. A combination fluid switch for controlling fluid flow through a plurality of fluid flow paths, comprising: (a) a plurality of fluid inputs; (b) a first two-position valve having respective fluid flow paths through which fluid from the plurality of fluid inputs is directed; (c) a second two-position valve having respective fluid flow paths through which fluid from the plurality of fluid inputs is directed; and (d) a plurality of fluid outputs; The combination fluid switch comprises: a fluid flow from a first fluid input to a first fluid output when the first two-position valve is in an open position and the second two-position valve is in an open position; a fluid flow from a second fluid input to a second fluid output when the first two-position valve is in a closed position and the second two-position valve is in an open position; a fluid flow from a third fluid input to a third fluid output when the first two-position valve is in an open position and the second two-position valve is in a closed position; and a combination fluid switch configured to allow fluid flow from a fourth fluid input to a fourth fluid output when the first two-position valve is in a closed position and when the second two-position valve is in a closed position.

2. The combination fluidic switch of claim 1 , wherein the fluid input comprises a tube.

3. The combination fluid switch of claim 1 or 2, wherein the fluid flow path comprises a tube.

4. The combination fluid switch of claim 1 , wherein the two-position valve is a trumpet valve.

5. The combination fluidic switch of claim 1 , wherein the fluid output comprises a tube.

6. 10. The combination fluid switch of claim 1, wherein said fluid input, said fluid flow path, and said fluid output comprise at least four tubing lines.

7. The combination fluidic switch of claim 1 , wherein each of said fluid inputs is supplied from a single fluid source.

8. 10. The combination fluid switch of claim 1, wherein fluids within the fluid flow paths are not allowed to mix.

9. 1. An automated biological production system comprising: (a) a sealable housing; (b) a cassette contained within the sealable housing, the cassette comprising: i. a cell culture chamber; ii. A combination fluid switch according to claim 1; iii. a pumping system fluidly connected to the cell culture chamber and the combination fluid switch; iv. one or more of a temperature sensor, a pH sensor, a glucose sensor, a lactose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor; v. An automated biological production system comprising: a cassette comprising a mechanism for automatically regulating one or more of temperature, pH level, glucose level, lactose level, oxygen level, carbon dioxide level, and optical density.

10. 10. The automated biological production system of claim 9 configured to produce cells.

11. 10. The automated biological production system of claim 9, further comprising a magnetic cell separation device.

12. 10. The automated biological production system of claim 9, further comprising an electroporation device.

13. 10. The automated biological production system of claim 9, wherein the automated biological production system comprises at least 16 fluid flow paths.

14. 10. The automated biological production system of claim 9, wherein the automated biological production system comprises at least 20 fluid flow paths.

15. 1. A combination fluid switch, comprising: (a) a housing having two opposing sides, each side having four openings therethrough; (b) two two-position valves disposed within the housing, each valve having four openings therethrough; the openings in the side and the openings in the two-position valve are configured to receive tubes therethrough to create four flow paths within the combination fluid switch; A combination fluid switch, wherein the two-position valve is movable within the housing to allow fluid flow through only one flow path at a time.

16. The combination fluid switch of claim 15 , wherein the two-position valve is slidable within the housing.

17. The combination fluid switch of claim 15 , wherein the two-position valve is rotatable within the housing.

18. The combination fluid switch of any one of claims 15 to 17, further comprising four tubing lines passing through the four openings in the side and the four openings in the two-position valve.

19. 1. A combination fluid switch, comprising: (a) a support base comprising two raised portions and two recessed portions, the two raised portions comprising a plurality of compartments extending above the support base configured to allow tubes to pass therethrough to create four fluid flow paths, the two recessed portions each comprising four stationary compression members extending above the support base; (b) two two-position valves having openings to allow the stationary compression members to pass therethrough, the valves further comprising four movable compression members configured to compress against complementary stationary compression members on the support base, thereby constricting a tube between the movable compression members and the complementary stationary compression members; A combination fluid switch, wherein the two-position valve is configured to slide along the support base and to allow fluid flow through only one flow path at a time.

20. 1. A combination fluid switch, comprising: (a) at least three stationary compression members; (b) a first two-position valve having two movable compression members, the two movable compression members being integrated into the first two-position valve; and (c) a second two-position valve having three movable compression members, the three movable compression members being integrated into the second two-position valve; the movable compression members of the first and second two-position valves are configured to compress against complementary stationary compression members, thereby constricting a tube between the movable compression member and the complementary stationary compression member; A combination fluid switch, wherein the two-position valve is configured to slide and allow fluid flow through one flow path at a time.

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