Impeller and sparger assemblies for bioprocessing system

Abstract

To provide impeller and / or sparger assemblies that provide increased oxygen transfer rates and kLa values in a bioreactor system to support increased cell culture cell densities.SOLUTION: A sparger assembly for a bioprocessing system includes a base plate and at least one aeration manifold removably connected to the base plate. Each aeration manifold includes at least one inlet for receiving a gas, and a plurality of gas outlet openings for delivering the gas to a fluid within the bioprocessing system. An impeller assembly for a bioprocessing system includes a hub, and at least one blade operatively connected to the hub. The at least one blade includes a first portion connected to the hub and extending generally vertically, and a second portion extending at an upward angle from the first portion.SELECTED DRAWING: Figure 10

Description

[Technical field]

[0001] FIELD OF THE DISCLOSURE

[0002] Embodiments of the present invention relate generally to bioprocessing systems and methods, and more particularly to an impeller and sparger assembly for a single-use biological reaction system. [Background technology]

[0002] A variety of vessels, devices, elements, and unit operations are known for carrying out biochemical and / or biological processes and / or manipulating the liquids and other products of such processes. To avoid the time, expense, and difficulty associated with sterilizing vessels used in biopharmaceutical manufacturing processes, single-use or disposable bioreaction bags and single-use mixing bags are used as such vessels. By way of example, biological materials (e.g., animal and plant cells), including, for example, mammalian, plant, or insect cells, and microbial cultures, can be processed using disposable or single-use mixers and bioreactors.

[0003] Increasingly, single-use or disposable containers are being used in the biopharmaceutical industry. Such containers can be flexible or collapsible plastic bags supported by an outer rigid structure such as a stainless steel shell or container. The use of sterilized disposable bags eliminates the time-consuming step of cleaning the container and reduces the chance of contamination. The bag can be positioned within the rigid container and filled with the desired fluids for mixing. A stirring assembly located within the bag is used to mix the fluids. Existing stirrers are either top-driven (having a shaft that extends downward into the bag and on which one or more impellers are mounted) or bottom-driven (having an impeller located at the bottom of the bag that is driven by a magnetic drive system or motor located outside the bag and / or container). Most magnetic stirring systems include a rotating magnetic drive head on the outside of the bag and a rotating magnetic stirrer (also referred to in this context as an "impeller") within the bag. The movement of the magnetic drive head allows for torque transmission, thereby allowing rotation of the magnetic stirrer, causing the stirrer to mix the fluids within the container. Magnetic coupling of the agitator inside the bag to a drive system or motor external to the bag and / or bioreactor vessel eliminates contamination issues, allows for a fully enclosed system, and prevents leaks. A magnetically coupled system can also eliminate the need to have a seal between the drive shaft and the vessel, since the drive shaft does not need to penetrate the wall of the bioreactor vessel to mechanically rotate the agitator.

[0004] Depending on the fluid being processed, the bioreactor system may include several fluid lines and different sensors, probes, and ports that are coupled to the bag for monitoring, analysis, sampling, and liquid transfer. For example, a sampling port is typically located at the bottom of the disposable bag and container, and a sampling line can be connected to the bag for sampling and draining from the bag. Existing bioreactor systems also typically utilize a sparger to introduce a controlled amount of a particular gas or combination of gases into the bioreactor. The sparger outputs small bubbles into the liquid to agitate the gas and / or dissolve the gas into the liquid. The delivery of gas through the sparger helps mix the materials and maintain a homogenous environment throughout the interior of the bag, which may be essential for cell growth in the bioreactor. Ideally, the sparger and the agitator are in close proximity to ensure optimal distribution of the gas throughout the container.

[0005] A high performance bioreactor system must provide good bulk mixing in combination with efficient gas dispersion to achieve large gas surface area and bubble size distribution, and therefore provide high oxygen transfer rates and kLa values ​​(volumetric mass transfer coefficient that represents the efficiency with which oxygen can be delivered to a bioreactor for a given set of operating conditions) that are desired in enhanced cell culture and / or microbial applications. Conventional solutions to achieve high kLa values ​​employ multiple impellers mounted on a single shaft. However, for single-use bioreactors, the use of multiple impellers results in a bulky appearance of disposable bags that cannot be folded efficiently. Furthermore, longer shafts with multiple impellers require stabilization, which increases the complexity and cost of the vessel and bag design, making the bag installation more cumbersome and difficult to use. Summary of the Invention [Problem to be solved by the invention]

[0006] In view of the above, there is a need for an impeller and / or sparger assembly that provides increased oxygen transfer rates and kLa values ​​in bioreactor systems to support increased cell culture cell densities. [Means for solving the problem]

[0007] In a first aspect, a sparger assembly for a bioprocessing system includes a base plate and at least one aeration manifold removably coupled to the base plate, each aeration manifold including at least one inlet for receiving gas and a plurality of gas outlet openings for delivering gas to a fluid in the bioprocessing system.

[0008] In a second embodiment, a bioprocessing system includes a vessel, a flexible bioprocessing bag positionable within the vessel, and a sparger assembly positioned at the bottom of the flexible bioprocessing bag. The sparger assembly includes a base plate and at least one aeration manifold removably coupled to the base plate. Each aeration manifold includes at least one inlet for receiving gas and at least one gas outlet opening for delivering gas to a fluid within the flexible bioprocessing bag.

[0009] In a third embodiment, a sparger assembly for a bioprocessing system comprises a base plate, at least one aeration manifold removably coupled to the base plate and supported in an elevated position relative to the base plate, each aeration manifold having at least one inlet for receiving gas and at least one gas outlet opening for delivering gas to a fluid in the bioprocessing system, and a cooperating mounting arrangement enabling coupling of an impeller of the bioprocessing system to the sparger assembly in close association with the aeration manifold.

[0010] In a fourth embodiment, an impeller assembly for a bioprocessing system includes a hub and at least one blade operably coupled to the hub, the at least one blade including a first portion coupled to the hub and extending generally vertically and a second portion extending at an upward angle from the first portion.

[0011] In a fifth embodiment, an impeller assembly for a bioprocessing system includes a hub having a central axis and a plurality of blades extending from the hub, at least one of the blades being oriented at one of a lead angle or a lag angle relative to a radial line extending from the central axis of the hub.

[0012] In a sixth embodiment, an impeller assembly for a bioprocessing system includes a hub and a plurality of blades extending from the hub, each blade having a leading edge and a trailing edge, at least one of the blades including an array of slots or openings in a leading edge of the blade.

[0013] The invention will be more readily understood from reading the following description of non-limiting embodiments, with reference to the accompanying drawings, in which: [Brief description of the drawings]

[0014] [Figure 1] FIG. 1 is a front elevation view of a biological reaction system in accordance with certain embodiments of the present invention. [Diagram 2] FIG. 2 is a simplified cross-sectional side elevation view of the biological reaction system of FIG. 1. [Diagram 3] FIG. 2 is a perspective view of a sparger assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 4] FIG. 2 is a perspective view of a sparger assembly according to certain embodiments of the present invention. [Diagram 5] FIG. 2 is a perspective view of a sparger assembly according to certain embodiments of the present invention. [Figure 6] FIG. 2 is a perspective view of a sparger assembly according to certain embodiments of the present invention. [Figure 7]FIG. 2 is a perspective view of a sparger assembly according to certain embodiments of the present invention. [Figure 8] FIG. 2 is a perspective view of a sparger assembly according to certain embodiments of the present invention. [Figure 9] FIG. 2 is a perspective view of a sparger assembly according to a particular embodiment of the present invention, shown with an impeller assembly mounted thereon. [Figure 10] FIG. 10 is a top plan view of the sparger assembly of FIG. [Figure 11] FIG. 2 is a perspective view of an impeller assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 12] FIG. 12 is a plan view from above of the impeller assembly of FIG. [Figure 13] FIG. 12 is a side elevational view of the impeller assembly of FIG. [Figure 14] FIG. 14 is an enlarged detail view of area A in FIG. 13. [Figure 15] FIG. 2 is a perspective view of an impeller assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 16] FIG. 2 is a perspective view of an impeller assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 17] FIG. 2 is a perspective view of an impeller assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 18] FIG. 18 is a schematic diagram of the impeller assembly of FIG. [Figure 19] FIG. 2 is a perspective view of an impeller assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 20] FIG. 2 is a perspective view of an impeller assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 21] FIG. 2 is a schematic diagram of an arrangement of openings in a sparger element / aeration manifold according to certain embodiments of the present invention. [Figure 22] FIG. 2 is a plan view from above of one arrangement of an aeration manifold of a sparger assembly in accordance with certain embodiments of the present invention. [Diagram 23] FIG. 2 is a perspective view of a sparger assembly according to certain embodiments of the present invention. [Figure 24] FIG. 24 is a side elevational view of the sparger assembly of FIG. 23 shown in use on a flexible biological reaction bag. [Diagram 25] FIG. 2 is a side view of an impeller assembly for use with the biological reaction system of FIG. 1, in accordance with certain embodiments of the present invention. [Figure 26] FIG. 26 is a perspective view of the impeller assembly of FIG. [Figure 27] FIG. 2 is an exploded perspective view of a sparger assembly according to certain embodiments of the present invention. [Figure 28] FIG. 28 is a top view of the sparger assembly of FIG. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Reference will now be made in detail to the exemplary embodiments of the present invention, examples of which are illustrated in the accompanying drawings. Wherever possible, the same reference numbers are used throughout the drawings to refer to the same or like parts.

[0016] As used herein, the terms "flexible" or "foldable" refer to structures or materials that are pliable or can bend without cracking, and can also refer to materials that are compressible or expandable. An example of a flexible structure is a bag formed from a polyethylene film. The terms "rigid" and "semi-rigid" are used interchangeably herein to describe structures that are "non-collapsible", i.e., structures that do not fold, collapse, or deform under normal forces to substantially shorten their elongated dimensions. Depending on the context, "semi-rigid" can also refer to structures that are more flexible than a "rigid" element, such as a bendable tube or conduit, but still do not collapse in the longitudinal direction under normal conditions and forces.

[0017] The term "vessel" is used herein and may refer to flexible bags, flexible containers, semi-rigid containers, rigid containers, or flexible or semi-rigid piping. The term "vessel" as used herein is intended to cover bioreactor containers having walls or portions of walls that are flexible or semi-rigid, single-use flexible bags, and other containers or conduits commonly used in biological or biochemical processes, including, for example, cell culture / purification systems, mixing systems, media / buffer preparation systems, filtration / purification systems, such as chromatography and tangential flow filtration systems, and their associated flow paths. As used herein, the term "bag" refers to flexible or semi-rigid containers or containers that are used, for example, as bioreactors or mixers for the contents therein.

[0018] As used herein, the terms "removably connected" or "removably coupled" mean that the aeration manifold / sparger element and base plate are connected in such a manner that they may be easily connected and / or removed to allow for easy user custom fabrication of the sparger assembly without special tools. Stated another way, "removably connected" is the opposite of "permanently connected."

[0019] Embodiments of the present invention provide a biological reaction system and a sparger assembly for the biological reaction system. In certain embodiments, the sparger assembly for the bioprocessing system comprises a base plate and at least one aeration manifold coupled to the base plate in a spaced apart vertical relationship relative to the base plate. Each aeration manifold comprises at least one inlet for receiving gas and a plurality of gas outlet openings for delivering gas to a fluid in the bioprocessing system.

[0020] Embodiments of the present invention provide a biological reaction system and an impeller assembly for the biological reaction system. In some embodiments, the impeller assembly for the bioprocessing system comprises a hub and at least one blade operably coupled to the hub. The at least one blade comprises a first portion coupled to the hub and extending generally vertically and a second portion extending at an upward angle from the first portion.

[0021] 1 and 2, a bioreactor system 10 according to some embodiments of the present invention is shown. The bioreactor system 10 includes a generally rigid bioreactor vessel or support structure 12 mounted on a base 14 having a plurality of legs 16. The vessel 12 may be formed, for example, from stainless steel, polymers, composites, glass, or other metals and may be cylindrical in shape, although other shapes may be utilized without departing from the broader aspects of the present invention. The vessel 12 may be provided with a lifting assembly 18 that provides support for a single-use flexible bag 20 that is disposed within the vessel 12. The vessel 12 may be of any shape or size so long as it is capable of supporting the single-use flexible bioreactor bag 20. For example, according to some embodiments of the present invention, the vessel 12 may be capable of receiving and supporting a 10-2000 L flexible or collapsible bioprocess bag assembly 20.

[0022] The vessel 12 may include one or more visual windows 22 that allow visualization of the fluid level in the flexible bag 20, and a window 24 positioned in a lower region of the vessel 12. The window 24 allows access to the interior of the vessel 12 for insertion and positioning of various sensors and probes (not shown) within the flexible bag 20, as well as for connecting one or more fluid lines to the flexible bag 20 for fluids, gases, etc., to be added to or withdrawn from the flexible bag 20. Sensors / probes and controls for monitoring and controlling critical process parameters include, for example, any one or more and combinations of temperature, pressure, pH, dissolved oxygen (DO), dissolved carbon dioxide (pCO2), mixing ratios, and gas flow rates.

[0023] With particular reference to FIG. 2, a schematic side elevation cross-sectional view of a bioreaction system 10 is shown. As shown therein, a single-use flexible bag 20 is disposed within and held by the container 12. In an embodiment, the single-use flexible bag 20 is formed from a suitable flexible material, such as a homopolymer or copolymer. The flexible material can be USP Class VI approved, for example, silicone, polycarbonate, polyethylene, and polypropylene. Non-limiting examples of flexible materials include polymers such as polyethylene (e.g., linear low density polyethylene and very low density polyethylene), polypropylene, polyvinyl chloride, polyvinyl dichloride, polyvinylidene chloride, ethylene vinyl acetate, polycarbonate, polymethacrylate, polyvinyl alcohol, nylon, silicone rubber, other synthetic rubbers, and / or plastics. In some embodiments, the flexible material can be a laminate of several different materials, such as, for example, laminates of Fortem™, Bioclear™ 10, and Bioclear 11 available from GE Healthcare Life Sciences. A portion of the flexible container may comprise a substantially rigid material, such as a rigid polymer, e.g., high density polyethylene, metal, or glass. The flexible bag may be supplied pre-sterilized, such as using gamma irradiation.

[0024] The flexible bag 20 includes an impeller 28 attached to a magnetic hub 30 at the bottom center of the interior of the bag, and the impeller 28 rotates on an impeller plate 32 also positioned at the bottom of the interior of the bag 20. Together, the impeller 28 and hub 30 (and in some embodiments the impeller plate 32) form an impeller assembly. A magnetic drive 34 external to the container 12 provides the motive force to rotate the magnetic hub 30 and impeller 28 to mix the contents of the flexible bag 20. While FIG. 2 shows the use of a magnetically driven impeller, other types of impellers and drive systems are possible, including a top driven impeller.

[0025] In certain embodiments, the impeller plate 32 may be configured as a sparger assembly used to introduce certain gases or air into the fluid in the bag 20 to agitate and / or dissolve the air or gas into the fluid. Thus, in some embodiments, the impeller, sparger, and their components form a combined impeller / sparger assembly. In other embodiments, the sparger assembly and impeller assembly may be separate and / or individual components. In either implementation, the sparger assembly and impeller assembly are in close proximity to ensure optimal distribution of gas throughout the bag 20, as discussed in detail below. As discussed below, it is contemplated that the sparger assembly (which may also serve as an impeller plate supporting the impeller) may take one of a variety of configurations.

[0026] For example, Figure 3 illustrates a particular embodiment of a sparger assembly 100 that may be utilized with the flexible bag 20 and bioreactor / bioprocessing system 10. As illustrated, the sparger assembly 100 includes a base plate 110 and a plurality of aeration passages, hollow aeration elements, or hollow aeration manifolds 112, 114 coupled to the base plate 110. In some embodiments, the aeration manifolds 112, 114 include a plurality of feet 116 that are received in corresponding support rods or mounting posts 118 of the base plate 110 for being supported in a vertically spaced apart relationship (i.e., elevated upwardly) relative to the base plate 110. In certain embodiments, the aeration manifolds 112, 114 and base plate 110 may be manufactured as an integral, single component. In other embodiments, the aeration manifolds 112, 114 may be manufactured as separate components that may be removably coupled to the base plate 110 through snap fastenings, clips, screws, or other connection means using feet 116 and posts 118. As shown in FIG. 3, each of the aeration manifolds 112, 114 may be arc shaped. In some embodiments, as shown in FIG. 3, the aeration manifolds 112, 114 may be semicircular arcs with multiple gas outlet openings or apertures 120 on their upper surfaces. In certain embodiments, the gas outlet openings 120 may be small holes in a porous frit. The aeration manifolds 112, 114 may also include one or more pipe couplers 122 forming inlets configured for mating connection with gas supply piping (not shown) to deliver gas to the aeration manifolds 112, 114. In some embodiments, the pipe connector 122 is a hose barb connector, although other connector types known in the art may be utilized without departing from the broader aspects of the present invention.

[0027] As shown in Figures 27 and 28, the aeration manifold 111 may be assembled from an inlet chamber unit 113 sealed to a perforated plate 119 via a gasket 117 and held together by a frame 121. The inlet chamber unit suitably has an inlet chamber 115 defined by a side wall 131 and a bottom wall 133, which is in fluid communication with a pipe connector 122 for a supply of gas that is evenly distributed through the inlet chamber to the perforated plate. The inlet chamber unit may be mounted to a base plate, for example, via feet 116 mounted with snap-fastening elements for forming a snap-fit ​​connection with columns in the base plate. The perforated plate may then be sealed to the inlet chamber via a gasket 117 in sealing abutment with the side wall 131 and a peripheral portion 135 of the perforated plate. The gasket 117 is shown as a separate detail, but may be molded integrally with the side wall 131 of the inlet chamber unit 113. The peripheral portion 135 can be non-porous, while the remainder of the plate is porous, with either an array of parallel small holes or a three-dimensionally connected porous network as in a porous frit. Alternatively, the entire plate is porous. Sealing pressure across the perforated plate and gasket can be applied by a frame that is connected to the inlet chamber unit, for example by a number of snap-fit ​​connections. The snap-fit ​​connections can, for example, comprise cantilevers 123 that extend from the periphery of the frame and engage with protrusions or lips 125 on the sidewalls of the inlet chamber unit. Alternatively, cantilevers extending from the inlet chamber unit can engage with protrusions / lips on the frame. The frame can further comprise one or more support members 127 extending across the frame to limit pressure bulging of the perforated plate.

[0028] In certain embodiments, the gas outlet openings 120 may all be the same size. In other embodiments, the gas outlet openings 120 of the first aeration manifold 112 may be different sizes than the gas outlet openings 120 of the second aeration manifold 114. For example, the gas outlet openings 120 of the first aeration manifold 112 may be smaller than the gas outlet openings 120 of the second aeration manifold 114. Thus, in such implementations, the first aeration manifold 112 with the relatively small gas outlet openings 120 generating relatively small bubbles may be utilized to supply oxygen, while the second aeration manifold 114 with the relatively large gas outlet openings 120 generating relatively large bubbles may be particularly suited to removing or scavenging CO2 with air, for example. If a porous frit is utilized, the openings / holes will not have the same size, but the various aeration manifolds may have openings with the same or different average sizes.

[0029] With further reference to FIG. 3, the base plate 110 may include a mounting arrangement that allows for coupling of an impeller of a bioprocessing system to the sparger assembly in close relation to the aeration manifold. In some embodiments, the mounting arrangement is a vertically extending mounting shaft 124 that is centrally located between the two arc-shaped manifolds 112, 114. The shaft 124 is configured to receive a magnetic hub (e.g., hub 30) of an impeller (e.g., impeller 28) and to support the impeller in a position where the lower edge of the impeller blades is located just above the upper surface of the manifolds 112, 114. Although the embodiments described herein disclose the sparger assembly as having a mounting shaft for receiving the impeller assembly, other cooperating mounting configurations are possible. For example, the sparger assemblies disclosed herein may have recessed bearings or receiving structures configured to receive a shaft secured to the impeller. Other coupling configurations are possible without departing from the broader aspects of the invention, including any configuration that employs retaining elements to couple the impeller and base plate together.

[0030] In certain embodiments, the base plate 110 may further include an opening 126 or fitting for fluidly coupling with drain piping for draining or harvesting the contents of the flexible bag 20. Incorporating the impeller mounting shaft 124 and drain opening 126 into the base plate 110 facilitates positioning of the flexible bag 20 within the bioreactor vessel 12, as well as aligning the magnetic hub 30 with the magnetic drive system and the drain port in the flexible bag 20 with the drain piping that is connected to the bottom of the bioreactor vessel 12.

[0031] 4, a sparger assembly 200 is shown according to some embodiments of the present invention. As shown, the sparger assembly 200 comprises a base plate 210 and a plurality of, i.e., four, aeration passages or hollow aeration manifolds 212, 214, 216, 218 coupled to the base plate 210. In some embodiments, the aeration manifolds 212, 214, 216, 218 comprise a plurality of feet 220 that are received in corresponding support rods or mounting posts 222 of the base plate 210 so as to be supported in a vertically spaced apart relationship (i.e., elevated upwardly) relative to the base plate 210, as previously described. Also as previously described, the aeration manifolds and base plate may be manufactured as an integral, single component or as separate components that may be removably coupled to the base plate 210 using the feet 220 and posts 222 through snap fastenings, clips, screws, or other connection means.

[0032] 4, each of the aeration manifolds can be a quarter circular arc and can include a plurality of gas outlet openings or apertures 224 on an upper surface thereof. The aeration manifolds 212, 214, 216, 218 can also include one or more pipe couplers 226 forming inlets configured for mating connection with one or more gas supply piping, such as piping 228, 230, for delivering gas to the aeration manifolds. In a particular embodiment, the pipe couplers 226 are hose barb couplers, although other connector types known in the art may be utilized without departing from the broader aspects of the present invention.

[0033] As with the embodiment of FIG. 3, the gas outlet openings 224 of each aeration manifold may be the same size. In other embodiments, the size of the gas outlet openings 224 of at least one of the aeration manifolds may differ from the size of the gas outlet openings 224 of at least one other of the aeration manifolds. For example, in some embodiments, a first pair of opposing aeration manifolds, e.g., aeration manifolds 212, 214 on opposite sides of a circle formed by the arrangement of the manifolds on the base plate 210, may have gas outlet openings 224 of a first size that differs from the size of the gas outlet openings 224 of a second pair of opposing aeration manifolds, e.g., aeration manifolds 216, 218 on opposite sides of a circle formed by the arrangement of the manifolds on the base plate. As previously disclosed, aeration manifolds with smaller gas outlet openings can be utilized to supply oxygen, while aeration manifolds with larger gas outlet openings can be utilized to remove or scavenge CO2 with air, for example.

[0034] In certain embodiments, an immediately adjacent pair of aeration manifolds, e.g., aeration manifolds 212, 216, may have gas outlet openings 224 of a first size, while another immediately adjacent pair of aeration manifolds, e.g., aeration manifolds 214, 218, may have gas outlet openings of a second size, where the second size is different from the first size. The configuration of base plate 210 and aeration manifolds 212, 214, 216, 218, and the selectively removable nature of the aeration manifolds, allow the configuration of sparger assembly 200 to be easily adjusted according to user preferences. In particular, this design allows for plug-and-play like functionality, such that a user can mount various combinations of aeration manifolds to base plate 210 to provide sparger assemblies of various configurations. For example, a user can easily install three aeration manifolds with smaller gas outlet openings 224 in combination with a single aeration manifold with a larger gas outlet opening 224 to increase oxygen delivery to the system if desired, or three aeration manifolds with larger gas outlet openings 224 in combination with a single aeration manifold with a smaller gas outlet opening 224 to increase CO2 removal without having to adjust the rate of gas delivery to the sparger assembly 200.

[0035] 3, the base plate 210 may include a vertically extending mounting shaft 232 centrally located between the aeration manifolds for receiving the impeller assembly. Additionally, as discussed above, the base plate 210 may include an opening 234 or fitting for fluidly coupling with exhaust piping for drainage or collection of the contents of the flexible bag 20.

[0036] Turning now to Figure 5, a sparger assembly 300 is shown according to some embodiments of the present invention. Sparger assembly 300 is similar in construction to sparger assembly 200 of Figure 4, with like numbers representing like parts. However, rather than each aeration manifold having a hose barb connector for connection to the gas supply piping, a T-shaped fitting 310 is utilized to fluidly interconnect two adjacent aeration manifolds (e.g., aeration manifold 212 and aeration manifold 216, and aeration manifold 214 and aeration manifold 218) and to connect gas supply piping 228, 230 to the aeration manifolds, respectively. In one implementation, the fluidly interconnected aeration manifolds may each have the same sized gas outlet openings 224. In another implementation, a first pair of interconnected aeration manifolds (e.g., aeration manifolds 212, 216) may have gas outlet openings 224 that are different in size than the gas outlet openings 224 of a second pair of interconnected aeration manifolds (e.g., aeration manifolds 214, 218). In yet other implementations, all of the aeration manifolds may have gas outlet openings 224 that are the same size.

[0037] Referring to Figure 6, a sparger assembly 400 is shown in accordance with certain embodiments of the present invention. The sparger assembly 400 is similar in construction to the sparger assembly 200 of Figure 4, with like numbers referring to like parts. However, rather than each aeration manifold having a hose barb connector for connection to the gas supply piping, an elbow fitting 410 is utilized to connect the gas supply piping 228, 230 to the aeration manifolds 212, 214, 216, 218, respectively. For example, an elbow fitting 410 may be utilized to connect the first gas supply piping 228 to the aeration manifolds 212, 216 and the second gas supply piping 230 to the aeration manifolds 214, 218. As previously described, some of the aeration manifolds may be configured with gas outlet openings 224 having a different size than those of the other aeration manifolds. In some embodiments, aeration manifolds that are connected to a common supply line may have gas outlet openings 224 that are the same size.

[0038] 3-6 depict sparger assemblies having two or four individual aeration manifolds, it is contemplated that the base plate may be manufactured with support posts 222 configured to receive three or more aeration manifolds of any partial circular shape (i.e., any section of a circle). Specifically, the sparger assembly may include any number of arc-shaped aeration manifolds that together form a broken (or unbroken) circular arc. In certain embodiments, the individual arc components may be separate components in a generally circular or annular arc that may be manufactured through additive manufacturing techniques. As such, the base plate allows the sparger assembly to be easily configured according to the user's preferences and adapted to the specific bioprocess being implemented in the biological reaction system 10. As previously discussed, the aeration manifolds may be configured for removable connection to the base plate to allow for easy custom fabrication of the sparger assembly.

[0039] 7, a sparger assembly 500 according to some embodiments of the present invention is shown. As shown, the sparger assembly 500 comprises a generally circular base plate 510 and an annular aeration manifold 512 removably coupled to the base plate 510. As with previously discussed embodiments, the aeration manifold 512 comprises a plurality of gas outlet openings 514 and is elevated above the base plate 510. In some embodiments, the aeration manifold 512 may comprise a plurality of feet 516 that are received by support rods or columns 518 in the base plate 510 to support the manifold 512 in a vertically spaced apart relationship relative to the base plate. The aeration manifold 512 may also comprise one or more pipe couplers 520 for coupling one or more gas supply pipes to the aeration manifold 512 in the manner previously described. Similar to the previously described embodiments, the base plate 510 may include a vertically extending mounting shaft 522 centrally located in the aeration manifold 512 for receiving the impeller assembly.

[0040] 8, a sparger assembly 600 according to certain embodiments of the present invention is shown. The sparger assembly 600 comprises a base plate 610 and a pair of nested aeration manifolds 612, 614 coupled to the base plate 610. As with the previously discussed embodiments, each aeration manifold 612, 614 comprises a plurality of gas outlet openings 616 and is elevated above the base plate 610 (e.g., supported on protruding posts 618 extending upwardly from the base plate 610). In certain embodiments, the aeration manifolds 612, 614 are removably coupled to the base plate 610 and comprise pipe couplers (not shown) for connecting one or more gas supply pipes (not shown) to the aeration manifolds 612, 614 in the manner previously described. Similar to previously described embodiments, the base plate 610 may include a vertically extending mounting shaft 620 centrally located in the aeration manifolds 612, 614 for receiving the impeller assembly of the biological reaction system 10. Additionally, the base plate 610 may include an opening 622 or fitting for fluidly coupling with drain piping for drainage or collection of the contents of the flexible bag 20.

[0041] 8, the aeration manifolds 612, 614 may have a pleated or sprocket-like shape. Specifically, in some embodiments, the outer aeration manifold 612 may have an inner periphery that is generally sprocket-shaped, and the inner aeration manifold 614 may have an outer periphery that is also generally sprocket-shaped. The inner aeration manifold 614 may be sized and oriented such that the "teeth" or crests 624 of the inner aeration manifold 614 are received in corresponding recesses or grooves 626 in the outer aeration manifold 612. In some embodiments, the gas exit openings 616 of the aeration manifolds 612, 614 may be the same or different sizes.

[0042] 9 and 10 show a particular embodiment of a sparger assembly 700 having a base plate 710 and a plurality of aeration manifolds supported thereon in an elevated or spaced apart vertical relationship relative to the base plate 710, according to some embodiments of the present invention. As shown therein, the aeration manifolds may comprise a plurality of outer arcuate aeration manifolds 712 and a plurality of inner arcuate aeration manifolds 714 nested with the outer aeration manifolds 712 or positioned at an inner radial position of the outer aeration manifolds 712. The aeration manifolds 712, 714 each include at least one gas outlet opening 716, the function of which has been previously described. The aeration manifolds 712, 714 are supported in an elevated position above the base plate 710 by a plurality of posts or protrusions (not shown), also as previously described.

[0043] In certain embodiments, the inner aeration manifold 714 and the outer aeration manifold are elevated above the support plate 710 at substantially the same distance (e.g., less than 5%, such as less than 1% difference in distance to the support plate). In certain embodiments, as best shown in FIG. 9, the inner aeration manifold 714 is positioned closer to the top surface of the base plate 710 than the outer aeration manifold 712. In this regard, the outer aeration manifold is elevated above the base plate 710 to a greater degree than the inner aeration manifold. This configuration allows the inner aeration manifold 714 to be positioned below the vanes of the impeller assembly 740, which is received on the impeller support shaft 718 (via the hub 744), and allows gas from the inner aeration manifold 714 to be discharged through the gas exit openings 716 below the vanes 742 of the impeller assembly 740. As shown in FIG. 10 , gas from the outer aeration manifold 712 can be discharged through gas exit openings 716 at a radial location outside the blades 742 of the impeller assembly 740 due to the positioning of the outer aeration manifold 712 radially outside the impeller blades.

[0044] Although the sparger assemblies of the present invention have been previously described as having sparger elements / aeration manifolds that are arc or arc shaped and arranged in a manner to form a circle or a portion of an arc, the present invention is not so limited in this respect. Specifically, the aeration manifolds themselves may have any shape desired (e.g., rectangular, triangular, oval, etc.) and may be arranged in an annular, circular, rectangular, or any polygonal shape. Other arrangements of the aeration manifolds in the base plate are also possible. For example, FIG. 22 shows a sparger assembly 750 having an aeration manifold 752 that is generally rectangular in shape and removably mounted in a base plate 754 to form a generally rectangular arrangement. In any embodiment, each aeration manifold may be separately or individually connected to one or more supplies of gases such that multiple gases may be delivered to each aeration manifold section as desired.

[0045] 23 and 24, a sparger assembly 760 is shown according to some embodiments of the present invention. However, rather than having an aeration manifold mounted in vertically spaced apart relationship to a base plate, the sparger assembly 760 comprises a base plate 762 having a hub (e.g., magnetic hub 30) and a sparger element or aeration manifold 764 extending radially from the hub 30. Although not mounted on a planar portion of the base plate 762, the aeration manifold is vertically spaced apart from the base plate. The aeration manifold 764 has gas exit openings 766, holes, or perforations that allow for dispersion of gas into the interior of the flexible biological reaction bag 20, as previously described. The aeration manifold 764 may be removably coupled to the hub 30, although in some embodiments, the aeration manifold 764 may be permanently fixed to the hub 30. As shown in FIG. 24, and as previously discussed, the magnetic hub 30 may include magnets 768 that cooperate with magnets 770 of the impeller 28 to drive the impeller 28 in rotation.

[0046] In conjunction with the previously described embodiments, by providing a sparger assembly with an aeration manifold for gas distribution that is elevated from the base plate (or at least elevated above the bottom surface of the vessel), the sparger gas can be introduced into the bioreactor in close association with the impeller, which provides more efficient gas dispersion to achieve a large gas surface area and bubble size distribution. Furthermore, because the aeration manifold is removably coupled to the base plate, the sparger assembly can be widely configurable and adaptable to provide almost any gas distribution profile desired. In particular, the modular nature of the sparger assembly described herein (i.e., base plate and removable aeration manifold) allows for easy customization and creation of the sparger assembly, including customization of gas outlet heights, gas outlet opening locations, sparging "density," etc.

[0047] In any of the previously described embodiments, the interior of the aeration manifold may be designed for optimal flow distribution, such as, for example, with a manifold groove system that promotes reduced pressure drop. In some embodiments, various components of the sparger assembly, including the aeration manifold, may be manufactured through additive manufacturing, which may be used to transition from solid to porous materials with built-in fluid passages to reduce part count and provide ease of assembly. While the previously described embodiments disclose hollow aeration manifolds with gas outlet openings, the manifold may be comprised of a porous frit, where the openings for gas release are small holes in the porous frit.

[0048] In some embodiments, the pattern of openings, holes, or pores in the aeration manifold of the sparger described herein can be any regular geometric pattern or random pattern. In certain embodiments, one or more openings in the aeration manifold can be arranged in a pattern configured such that the spacing s between the openings, holes, or pores is greater than the diameter of the bubbles generated by the openings, holes, or pores of diameter d. Having spacing between the openings, holes, or pores that is greater than the diameter of the bubbles helps prevent adjacent bubbles from coalescing because the bubbles do not contact each other at the surface of the sparger element / aeration manifold. The diameter of the bubbles generated by an opening, hole, or pore of a particular diameter not only depends on the diameter of the hole or pore, but is also greatly influenced by factors such as the surface energy of the material from which the sparger is constructed, and also depends on the physical and chemical properties of the liquid in which the bubbles are created, as this affects the surface tension of the air / liquid interface at the bubble surface.

[0049] Referring to FIG. 21, an example of a geometric pattern for the location of openings, holes, or perforations on the surface of an aeration manifold is shown. As shown in FIG. 21, the number of holes (e.g., holes 224) in a sparger element / aeration manifold (e.g., aeration manifold 112) is maximized by arranging the holes in an equilateral triangular pattern, where the holes are located at the apex of the triangle. This pattern may also be referred to as a hexagonal pattern. This pattern maximizes the number of holes that can be created in a sparger element with a particular surface area. In the equilateral triangular pattern of FIG. 21, every opening, hole, or perforation is equidistant from adjacent openings, holes, or perforations. Other geometric patterns, such as a simple rectangular grid, may be used without departing from the broader aspects of the invention. When holes or perforations are located at the corners of a rectangular grid, adjacent holes in the sparger element are located at two different distances, at the desired horizontal and vertical distance, and at a longer distance on the diagonal. Thus, for a particular desired minimum spacing between adjacent holes, the spacing of the holes on the diagonal will be a distance greater than the desired minimum. Such a rectangular pattern will result in a smaller number of holes in a particular surface area of ​​the sparger element than would be the case for a more efficient equilateral triangular pattern.

[0050] 11-18, various configurations of the impeller assembly of the bioreactor / bioprocessing system 10 are shown. With particular reference to FIGS. 11-14, in certain embodiments, the impeller assembly 800 comprises a hub 810 and at least one blade 812 extending radially from the hub 810. The hub 810 is rotatable about a vertical axis 814 extending through a center of the hub 810. In some embodiments, the hub 810 can be a magnetic hub configured to be driven by a magnetic drive system or motor (e.g., motor 34 of FIG. 2) positioned external to the flexible bag 20 and vessel 12.

[0051] Although the impeller assembly 800 is shown in FIGS. 11-14 as having three blades 812, it may have fewer than three blades (e.g., one blade or two blades) or more than three blades (e.g., four, five, or six blades) without departing from the broader aspects of the invention. The blades 812 may be equally spaced from one another around the hub 810. For example, the impeller assembly 800 may have three blades 812, and the blades 812 may be spaced 120° apart. The blades 812 each include a first portion 816 that is substantially vertical (e.g., deviating from vertical by less than 5°, such as less than 1°) and a non-vertical, non-horizontal, angled second portion 818 that extends upward from the first portion 816. While the first portion 816 and the second portion 818 are shown as being substantially planar (e.g., with a flatness tolerance of less than 5 mm, such as less than 1 mm), it is contemplated that in some embodiments, one or both of the first portion 816 and the second portion 818 of the vane 812 may have a curved or arcuate shape. As best shown in FIG. 13, the angled second portion 818 comprises a radiused portion 820 at the distal end of the vane 812. A radius 822 is also formed at the intersection between the vertical first portion 816 and the angled second portion 818.

[0052] With particular reference to FIGS. 13 and 14, impeller assembly 800 has a diameter d defined as the longest linear dimension from tip to tip. In certain embodiments, impeller diameter d may range from about one-quarter to about one-half the inner diameter of vessel 12. As best shown here, vertical first portion 816 and angled second portion form an angle α therebetween. In some embodiments, angle α is between about 100 degrees and about 180 degrees, such as between 120 degrees and 160 degrees. In certain embodiments, angle α is about 135 degrees (e.g., 130-140 degrees) such that angled second portion extends at an upward angle of about 45 degrees from horizontal.

[0053] As alluded to above, the impeller assembly 800 may be seated on the bottom of the flexible bag 20 in close association with the sparger assembly. For example, the impeller assembly 800 may be coupled to the base plate of one of the sparger assemblies disclosed herein such that the impeller blades 812 are in close association with the gas outlet openings of the sparger assembly. Through testing, it has been shown that the vertically straight portions 816 of the blades 812 of the impeller assembly 800 are particularly efficient at breaking up bubbles injected by the sparger assembly into the flexible bag 20, delivering significant power to the bioreactor system 10. Testing has also demonstrated that the angled portions 818 of the blades 812 facilitate mixing of the contents of the flexible bag 20. This combination of straight and angled blade sections therefore produces improved bubble breakage and efficient gas distribution (kLa) at optimal power consumption (i.e., without the need for higher power inputs or very high speed mixing that can cause shear damage and generate vortices that are harmful to cells).

[0054] In this regard, impeller assembly 800 optimizes high volume mixing and efficient gas distribution in a gas sparger to provide high oxygen transfer rates and kLa values, which are desirable in enhanced cell culture and / or microbial applications. In contrast to existing systems and devices, impeller assembly 800 achieves this performance while maintaining a relatively small profile (i.e., impeller assembly 800 remains bottom driven and located near the bottom of bag 20, allowing the bag to still easily collapse for storage and transport). This simple design also allows for easy user installation and configuration. Specifically, in some embodiments, impeller assembly 800 can be quickly and easily positioned on the mounting shaft of the sparger assembly base plate in the manner previously described.

[0055] Turning now to FIG. 15, an impeller assembly 850 is shown in accordance with certain embodiments of the present invention. As shown therein, the impeller assembly 850 includes a hub 852 and at least one vane 854 coupled to the hub 852. As in the embodiment of FIGS. 11-14, the hub 852 is rotatable about a vertical axis extending through a center of the hub 852. In some embodiments, the hub 852 may be a magnetic hub configured to be driven by a magnetic drive system or motor (e.g., motor 34 of FIG. 2) positioned external to the flexible bag 20 and container 12. In certain embodiments, the hub 852 may be formed as (or may be integral with) a generally flat disc 856 from which the vanes 854 extend.

[0056] The vanes 854 are substantially similar to the vanes 812 of the impeller assembly 800 of FIGS. 11-14, each including a first portion 858 that is substantially vertical (e.g., less than 5° deviation from vertical, such as less than 1°) and a non-vertical, non-horizontal, angled second portion 860 that extends upward from the first portion 858. Although the first portion 858 and the second portion 860 are shown as being substantially planar (e.g., with a flatness tolerance of less than 5 mm, such as less than 1 mm), it is contemplated that in some embodiments, one or both of the first portion 858 and the second portion 860 of the vane 854 may have a curved or arcuate shape. As shown in FIG. 15, in some embodiments, the vertical first portion 858 extends downward from the distribution disk 856, while the second portion 860 extends diagonally upward from the distribution disk 856. The vanes 854 may terminate at the outer periphery of the distribution disc 856 or may extend some way beyond such periphery, as shown in FIG.

[0057] Turning now to Figure 16, another impeller assembly 870 is shown according to some embodiments of the present invention. The impeller assembly 870 is substantially similar to the impeller assembly 850 of Figure 15, with like numbers representing like parts. However, as shown in Figure 16, the distribution disk 856 may additionally include radial slots 872 adjacent each (or at least some) of the vanes 854.

[0058] 25 and 26 show an impeller assembly 880 in which each vane 854, in addition to a substantially vertical first portion 858 and an angled second portion 860, further comprises a substantially horizontal (e.g., less than 5° deviation from horizontal, such as less than 1°) third portion 882 extending from a top end of the second portion. The third portion may have a distal end 884 that is wider than the proximal end 886. The third portion may even be substantially triangular with the distal end 884 forming the base of the triangle. The distal edges 858a, 860a, and 882a of the first, second, and third portions may be appropriately radiused to avoid potential damage to the walls of the flexible bioprocess bag.

[0059] Although the impeller assemblies 850, 870, 880 of Figures 15-16 and 25-26 have six blades, the impeller assemblies may have more or less than six blades, for example, three, four, five, seven, or eight blades, without departing from the broader aspects of the invention. In the embodiment of Figures 15 and 16, the vertical blade sections of the blades provide efficient radial liquid flow, while the angled blade sections allow axial fluid flow. Additionally, the distribution disk 856 functions to capture and thicken air / gas bubbles from the sparger assembly before dispersing them. As shown in Figure 16, slots 872 in the distribution disk 856 allow different bubble distribution patterns. These impeller assembly designs provide proper mixing and mass transfer of oxygen from the gas phase to the liquid phase, which is essential for cell culture for biopharmaceutical manufacturing, for example, at very large cell concentrations, where the demand for oxygen and uniform mixing is very high. Additionally, the impeller assembly disclosed herein efficiently disperses air bubbles from the sparger and provides efficient mixing without very high speed agitation which can cause shear damage and create harmful vortexes.

[0060] Turning now to Figure 17, there is shown an impeller assembly 900 according to some embodiments of the present invention. The impeller assembly 900 comprises a hub 910 and a number of blades 912, 914 attached to and extending radially outward from the hub 910. In certain embodiments, the hub 910 is a magnetic hub configured to be driven by an external magnetic drive system or motor, as previously discussed. Although Figure 17 shows the impeller assembly 900 having six blades 912, 914, the impeller assembly may have fewer or more than six blades without departing from the broader aspects of the present invention.

[0061] In some embodiments, one or more blades 912, 914 are coupled to the hub 910 at an angle offset from a radial line extending from the impeller shaft. For example, the blade 912 can be canted forward of a radial line extending from the impeller shaft with respect to a direction 916 of rotation of the impeller assembly 900, and the blade 914 can be canted backward of a radial line extending from the impeller shaft with respect to a direction 916 of rotation of the impeller assembly 900. As shown in FIG. 17, the blades may be alternately canted forward and backward. In such implementations, this blade configuration results in longer and shorter distances between the blade tips compared to the uniform distance between the blade tips without such canted or tilted blades. For example, the distance d1 between the tip of the backward oblique blade 914 (moving in the direction of rotation of the impeller assembly 900) and the tip of the next adjacent forward oblique blade 912 is increased compared to the distance between the blade tips when the blades are oriented along a radial line extending from the center of the hub 910. Also, the distance d2 between the tip of the forward oblique blade 912 (moving in the direction of rotation of the impeller assembly 900) and the tip of the next adjacent backward oblique blade 914 is decreased compared to the distance between the blade tips when the blades are oriented along a radial line extending from the center of the hub 910. In this regard, the impeller assembly 900 has alternating longer and shorter distances between the tips of the blades.

[0062] This inclination configuration of the vanes 912, 914 of the impeller assembly is shown more clearly in FIG. 18. As shown there, the alternating vanes 912 are oriented at a lead angle β1 relative to a precise radial line 918 extending from a central axis 920 of the impeller assembly 900. In contrast, the alternating vanes 914 are oriented at a lag angle β2 relative to a precise radial line 918 extending from a central axis 920 of the impeller assembly 900. In certain embodiments, the lead angle β1 of the vane 912 may be equal to the lag angle β2 of the vane 914. For example, in some embodiments, the lead angle β1 and the lag angle β2 may be between about 5 degrees and about 30 degrees. In some embodiments, the lead angle β1 and the lag angle β2 may be between about 5 degrees and about 10 degrees. In certain embodiments, the lead angle β1 and the lag angle β2 may be about 7 degrees, such as 6 to 8 degrees. In other embodiments, the lead angle β1 of vane 912 may be different from the delay angle β2 of vane 914. In still other embodiments, one or more of vanes 912 may have a different lead angle β1 than at least one other of vanes 912. Similarly, one or more of vanes 914 may have a different delay angle β2 than at least one other of vanes 914. It is contemplated that the number of vanes with a lead angle and the number of vanes with a delay angle may be the same or different.

[0063] In operation, the blades 912 oriented at a leading angle relative to a precise radial line 918 extending from the central axis 920 function to pull liquid inwardly toward the hub 910 in the direction of arrow B, as shown in FIG. 18. Conversely, the blades 914 oriented at a lagging angle relative to a precise radial line 918 extending from the central axis 920 function to push liquid away from the hub 910 in the direction of arrow C, as shown in FIG. 18. Thus, the impeller assembly 900 can be utilized to increase the efficiency of mixing, which can improve oxygen transfer within the bioreaction system 10. It is contemplated that the blade orientation / tilt aspects of the present invention can be employed in combination with existing blade shapes / forms / configurations known in the art to improve the mixing capabilities of an impeller.

[0064] Turning now to FIG. 19, an impeller assembly according to some embodiments of the present invention is shown. The impeller assembly 1000 comprises a hub 1010 and a number of blades 1012 mounted to the hub 1010. Although the impeller assembly 1000 of FIG. 19 has three blades 1012, fewer or more than three blades may be employed without departing from the broader aspects of the present invention. In certain embodiments, the impeller assembly 1000 is a marine impeller having arcuate or curved blades 1012. As shown in FIG. 19, in some embodiments, one or more of the blades 1012 comprise a number of slots 1014. In certain embodiments, the slots 1014 are generally vertically extending slots and are positioned at locations in the blades 1012 that are generally vertically aligned with locations in the sparger assembly where the sparger gas is discharged into the flexible bag 20. In some embodiments, the slot 1014 is formed in the forward or leading edge of the vane 1012 .

[0065] In use, the impeller assembly 1000 may be mounted to a mounting shaft of a sparger assembly, as discussed above. As indicated above, the slots 1014 are positioned such that as the vanes 1012 rotate, the slots 1014 pass closely over the gas exit openings in the sparger assembly.

[0066] Finally, referring to Figure 20, a similar impeller assembly 1100 is shown. However, rather than having slots in the leading edges of the vanes 1012, an array of dimples, holes, or openings 1110 may be formed in the leading edges of the vanes 1012. As with the embodiment of Figure 19, the openings 1110 are located in locations that generally correspond to the locations of the gas exit openings of the sparger assembly in which the impeller assembly 1100 is located.

[0067] It is also contemplated that slots or openings may be combined with any existing impeller design or configuration for a bioreactor system and with the impeller assembly configurations described herein. Utilizing an impeller with slots or openings in the area of ​​the impeller that passes closely over the gas outlet opening of the sparger assembly may increase the interfacial contact between the impeller blades and the fluid in the flexible bag 20. Thus, the impeller assemblies 1000, 1100 provide more efficient gas distribution in the gas sparger to provide the high oxygen transfer rates and kLa values ​​desired for enhancing cell culture without increasing the power requirements in the impeller drive system.

[0068] The impeller assembly and sparger assembly embodiments disclosed herein, and combinations thereof, provide various means of increasing the kLa (i.e., achieving more efficient gas distribution) of a biological reaction system to support enhanced cell culture and / or microbial applications. It is contemplated that the impeller assemblies disclosed herein may be utilized in combination with any existing sparger assembly. Similarly, the sparger assemblies disclosed herein may be utilized in conjunction with many existing impeller assemblies. Still further, it is contemplated that any of the impeller assemblies disclosed herein may be utilized in combination with any of the sparger assemblies also disclosed herein to provide both improved bulk mixing and efficient gas distribution. In this regard, the configuration of both the impeller assembly and sparger assembly of the present invention facilitates simple user operation or configuration of the combined impeller and sparger assembly. In particular, the impeller and / or sparger assemblies of the present invention can be easily manipulated (e.g., by interchanging aeration manifolds in the spargers and / or by connecting different impellers to the base plate of the sparger) to achieve almost any level of mass mixing or gas dispersion desired, depending on the particular cell culture or bioprocessing operation being performed in the bioprocessing system 10.

[0069] In some embodiments, a sparger assembly for a bioprocessing system comprises a base plate and at least one aeration manifold removably coupled to the base plate, each aeration manifold comprising at least one inlet for receiving gas and a plurality of gas outlet openings for delivering gas to a fluid in the bioprocessing system. In certain embodiments, the at least one aeration manifold is in a spaced apart vertical relationship to the base plate when coupled to the base plate. In some embodiments, the at least one aeration manifold can be annular in shape. In certain embodiments, the at least one aeration manifold is a two aeration manifold including a first aeration manifold and a second aeration manifold, the aeration manifolds being arranged in the form of a semicircular arc. In some embodiments, the at least one aeration manifold is a four aeration manifold, the aeration manifolds forming a quarter circular arc. In certain embodiments, the size of the gas outlet opening of at least one of the aeration manifolds is different from the size of the gas outlet opening of at least one other of the aeration manifolds. In some embodiments, the aeration manifolds are arranged on the base plate such that the size of the gas outlet opening of each aeration manifold is different from the size of the gas outlet opening of the immediately adjacent aeration manifold. In certain embodiments, a first pair of aeration manifolds are arranged on the base plate to form a first semicircle and a second pair of aeration manifolds are arranged on the base plate to form a second semicircle. The gas outlet openings of the first pair of aeration manifolds are a first size and the gas outlet openings of the second pair of aeration manifolds are a second size different from the first size. In some embodiments, the at least one aeration manifold is at least two aeration manifolds comprising a first aeration manifold and a second aeration manifold mounted coaxially with the first aeration manifold, and at least one of the first aeration manifold and the second aeration manifold has a pleated or sprocket-like periphery.In some embodiments, the at least one aeration manifold is a plurality of aeration manifolds, and at least one of the aeration manifolds is elevated a first distance above the base plate and at least one other of the aeration manifolds is elevated a second distance above the base plate, the first distance being greater than the second distance. In certain embodiments, the base plate comprises a shaft for receiving the impeller assembly and positioning the impeller assembly such that blades of the impeller assembly are positioned a distance above the at least one aeration manifold. In some embodiments, the base plate comprises an opening for mating with an exhaust port in the flexible bioprocess bag and an exhaust port of a support vessel that receives the flexible bioprocess bag.

[0070] In certain embodiments, the bioprocessing system includes a vessel, a flexible bioprocessing bag positionable in the vessel, and a sparger assembly positioned at the bottom of the flexible bioprocessing bag, the sparger assembly including a base plate and at least one aeration manifold removably coupled to and supported by the base plate. Each aeration manifold includes at least one inlet for receiving gas and at least one gas outlet opening for delivering gas to a fluid in the flexible bioprocessing bag. In some embodiments, the at least one aeration manifold is in a spaced apart vertical relationship to the base plate when coupled to the base plate. In certain embodiments, the at least one aeration manifold is annular in shape. In some embodiments, the at least one aeration manifold is a two aeration manifold including a first aeration manifold and a second aeration manifold, each of the aeration manifolds being arranged in the form of a semicircular arc. In certain embodiments, the at least one aeration manifold is four aeration manifolds, each of the aeration manifolds being approximately a quarter circular arc. In some embodiments, the size of the at least one gas outlet opening of at least one of the aeration manifolds is different than the size of the at least one gas outlet opening of at least one other of the aeration manifolds. In certain embodiments, the base plate comprises an opening for mating with an exhaust port in a flexible bioprocess bag and an exhaust port of a vessel.

[0071] In some embodiments, a sparger assembly for a bioprocessing system comprises a base plate, at least one aeration manifold removably coupled to the base plate and supported in an elevated position relative to the base plate, each aeration manifold having at least one inlet for receiving gas and at least one gas outlet opening for delivering gas to a fluid in the bioprocessing system, and a mounting arrangement that enables coupling of an impeller of the bioprocessing system to the sparger assembly in close relationship to the aeration manifold.

[0072] In some embodiments, an impeller assembly for a bioprocessing system comprises a hub and at least one blade operably coupled to the hub. The at least one blade comprises a first portion coupled to the hub and extending generally vertically and a second portion extending at an upward angle from the first portion. In certain embodiments, the first portion and the second portion are substantially planar (e.g., with a flatness tolerance of less than 5 mm, such as less than 1 mm). In some embodiments, the at least one blade is three blades. In certain embodiments, the second portion comprises a radiused distal end. In some embodiments, the hub comprises a generally planar disc, the disc comprising at least one slot adjacent each of the at least one blade. In some embodiments, the first portion and the second portion form an angle between about 100 degrees and about 180 degrees between them. In certain embodiments, the hub is a magnetic hub.

[0073] In certain embodiments, an impeller assembly for a bioprocessing system includes a hub having a central axis and a plurality of blades extending from the hub, at least one of the plurality of blades being oriented at one of a lead angle or a lag angle relative to a radial line extending from the central axis of the hub. In some embodiments, at least one of the plurality of blades is oriented at a lead angle relative to a first radial line extending from the central axis, and at least one other of the plurality of blades is oriented at a lag angle relative to a second radial line extending from the central axis. In certain embodiments, each of the plurality of blades is oriented at one of a lead angle or a lag angle relative to a respective radial line extending from the central axis of the hub, defining a lead blade or a lag blade of the plurality of blades. In some embodiments, the lead blades and the lag blades are positioned alternately in the direction of rotation of the hub. In certain embodiments, the lead angle is between about 5 degrees and about 30 degrees, and the lag angle is between about 5 degrees and about 30 degrees. In some embodiments, the distance between the tip of a leading blade and the tip of the next adjacent lagging blade, relative to the direction of rotation of the impeller assembly, is less than the distance between the tip of a lagging blade and the tip of the next adjacent leading blade. In certain embodiments, the impeller assembly has a maximum diameter that is between about one-quarter and about one-half the diameter of a bioreactor vessel in which the impeller assembly is configured to be positioned.

[0074] In some embodiments, an impeller assembly for a bioprocessing system includes a hub and a plurality of blades extending from the hub, each blade having a leading edge and a trailing edge. At least one of the blades includes an array of slots or openings at a leading edge of the blade. In some embodiments, the blade is a marine blade. In certain embodiments, the slot is a generally vertically extending slot. In some embodiments, each of the blades of the plurality of blades includes an array of slots or openings, respectively, at a leading edge of the blade. In certain embodiments, the array of slots or openings is located at a radial position on the blade that corresponds to a gas outlet of a sparger assembly of the bioprocessing system.

[0075] As used herein, elements or steps referred to in the singular and elements or steps preceded by the words "a" or "an" should be understood as not excluding a plurality of said elements or steps, unless expressly stated. Furthermore, references to "embodiments" of the invention are not intended to be interpreted as excluding the existence of additional embodiments that also incorporate the claimed features. Furthermore, unless expressly stated to the contrary, embodiments "comprising," "including," or "having" an element or elements having a particular property may include additional such elements not having that property. Any directional terms, such as "top," "bottom," "upper," "lower," "above," "below," "horizontal," "vertical," etc., refer to the directions as shown in the drawings, which are also the directions in the operating location of the bioprocessing system.

[0076] This written description uses examples to disclose some embodiments of the invention, including the best mode, and to enable one of ordinary skill in the art to practice embodiments of the invention, including making any device or system, using any device or system, and performing any incorporated methods. The patentable scope of the invention is defined by the claims, and may include other examples that occur to those of ordinary skill in the art. Such other examples are intended to be within the scope of the claims if they have structural elements that do not differ from the language of the claims, or if they include equivalent structural elements with insubstantial differences from the language of the claims.

[0077] [Section 1] A sparger assembly (100; 200; 300; 400; 500; 600; 700; 750; 760) for a bioprocessing system (10), comprising: Base plate (110;210;510;610;710;754) and at least one aeration manifold (112, 114; 212, 214, 218; 512; 612, 614) removably coupled to the base plate, each aeration manifold having at least one inlet for receiving gas and a plurality of gas outlet openings (120; 224; 514; 616; 716; 766) for delivering the gas to a fluid in the bioprocessing system; A sparger assembly comprising: [Section 2] 2. The sparger assembly of claim 1, wherein the at least one aeration manifold is in spaced vertical relationship to the base plate when coupled to the base plate. [Section 3] 3. The sparger assembly of claim 1 or 2, wherein the at least one aeration manifold has an annular shape. [Section 4] The at least one aeration manifold (111) comprises: an inlet chamber unit (113) having an inlet chamber (115) fluidly connected to a pipe connector (122); a perforated plate (119) sealed to the inlet chamber unit by a gasket (117); a frame (121) fixed to the inlet chamber unit and engaging the perforated plate to maintain the perforated plate and the gasket in sealing abutment with the inlet chamber unit; 4. The sparger assembly of any one of claims 1 to 3, comprising: [Section 5] 5. The sparger assembly of claim 4, wherein the frame is secured to the inlet chamber unit by a plurality of snap-fit ​​connections, such as a cantilever portion (123) extending from the frame engaging a protrusion (125) on the inlet chamber unit. [Section 6] 6. The sparger assembly of claim 4 or 5, wherein the inlet chamber unit comprises a plurality of feet (116) secured to posts in the base plate by snap-fit ​​connections (129). [Section 7] 7. The sparger assembly of any one of claims 1 to 6, wherein the at least one aeration manifold is a two aeration manifold including a first aeration manifold and a second aeration manifold, the aeration manifolds being arranged in a circular configuration on the base plate. [Section 8] 8. The sparger assembly of any one of claims 1 to 7, wherein the at least one aeration manifold is four aeration manifolds, the aeration manifolds being arranged in a circular configuration on the base plate. [Section 9] 9. The sparger assembly of claim 8, wherein the gas outlet opening of at least one of the aeration manifolds is a different size than the gas outlet opening of at least one other of the aeration manifolds. [Section 10] 10. The sparger assembly of any one of claims 7 to 9, wherein the aeration manifolds are positioned on the base plate such that the size of the gas outlet opening of each aeration manifold differs from the size of the gas outlet opening of an immediately adjacent aeration manifold. [Section 11] a first pair of the aeration manifolds are disposed on the base plate to form a first semicircle; a second pair of the aeration manifolds are disposed on the base plate to form a second semicircle; the gas outlet openings of the first pair of aeration manifolds are a first size; 11. The sparger assembly of any one of clauses 8 to 10, wherein the gas outlet openings of the second pair of aeration manifolds are a second size different than the first size. [Section 12] the at least one aeration manifold is at least two aeration manifolds comprising a first aeration manifold and a second aeration manifold mounted coaxially with the first aeration manifold; 3. The sparger assembly of claim 1 or 2, wherein at least one of the first aeration manifold and the second aeration manifold has a pleated or sprocket-like periphery. [Section 13] the at least one aeration manifold is a plurality of aeration manifolds; At least one of the aeration manifolds is elevated a first distance above the base plate; at least another of the aeration manifolds is elevated a second distance above the base plate; 13. The sparger assembly of any one of claims 1 to 12, wherein the first distance is greater than the second distance. [Section 14] 14. The sparger assembly of any one of claims 1 to 13, wherein the base plate receives an impeller assembly (28; 740; 800; 850; 870; 900; 1000; 1100) and comprises a shaft (124; 232; 522; 620; 718) for positioning the impeller assembly such that the blades (742; 812; 854; 912, 914; 1012) of the impeller assembly are positioned a distance above the at least one aeration manifold. [Section 15] 15. The sparger assembly of any one of claims 1 to 14, wherein the base plate includes an opening (234; 622) for mating with an outlet port in a flexible bioprocess bag (20) and an outlet port of a support vessel (12) that receives the flexible bioprocess bag. [Section 16] A container (12); a flexible bioprocessing bag (20) positionable within the vessel; a sparger assembly (100; 200; 300; 400; 500; 600; 700; 750; 760) positioned at the bottom of the flexible bioprocess bag, the sparger assembly comprising a base plate (110; 210; 510; 610; 710; 754) and at least one aeration manifold (112, 114; 212, 214, 218; 512; 612, 614) removably coupled to and supported by the base plate; A bioprocessing system (10) comprising: 1. A bioprocessing system, wherein each aeration manifold comprises at least one inlet for receiving a gas and at least one gas outlet opening (120; 224; 514; 616; 716; 766) for delivering said gas to fluid in said flexible bioprocess bag. [Section 17] 17. The bioprocessing system of clause 16, wherein the at least one aeration manifold is in spaced vertical relationship to the base plate when coupled to the base plate. [Section 18] 18. The bioprocessing system of clause 16 or clause 17, wherein the at least one aeration manifold has an annular shape. [Section 19] 19. The bioprocessing system of any one of clauses 16 to 18, wherein the at least one aeration manifold is a two aeration manifold including a first aeration manifold and a second aeration manifold, the aeration manifolds being arranged in a circular configuration on the base plate. [Section 20] 19. The bioprocessing system of any one of clauses 16 to 18, wherein the at least one aeration manifold is four aeration manifolds, the aeration manifolds being arranged in a circular configuration on the base plate. [Section 21] 21. The bioprocessing system of claim 19 or 20, wherein a size of the at least one gas outlet opening of at least one of the aeration manifolds differs from a size of the at least one gas outlet opening of at least one other of the aeration manifolds. [Section 22] 22. The bioprocessing system according to any one of claims 16 to 21, wherein the base plate comprises an opening (234; 622) for joining with an outlet port in a flexible bioprocess bag (20) and with an outlet port of the vessel (12). [Section 23] 23. The bioprocessing system of any one of clauses 16 to 22, wherein the sparger assembly is the sparger assembly of any one of clauses 1 to 15 or 24. [Section 24] A sparger assembly (100; 200; 300; 400; 500; 600; 700; 750; 760) for a bioprocessing system (10), comprising: Base plate (110;210;510;610;710;754) and at least one aeration manifold (112, 114; 212, 214, 218; 512; 612, 614) removably coupled to the base plate and supported in an elevated position relative to the base plate, each aeration manifold having at least one inlet for receiving gas and at least one gas outlet opening (120; 224; 514; 616; 716; 766) for delivering the gas to a fluid in the bioprocessing system; a mounting device (124; 232; 522; 620; 718) that allows for coupling of an impeller (28; 740; 800; 850; 870; 900; 1000; 1100) of the bioprocessing system to the sparger assembly in close association with the aeration manifold; A sparger assembly comprising: [Section 25] An impeller assembly (740; 800; 850; 870; 880; 900; 1000; 1100) for a bioprocessing system, comprising: Hub (30;810;852;910;1010) and at least one blade (742; 812; 854; 912, 914; 1012) operably coupled to said hub, said at least one blade having a first portion (816; 858) coupled to said hub and extending generally vertically, and a second portion (818; 860) extending at an upward angle from said first portion; An impeller assembly comprising: [Section 26] 26. The impeller assembly of clause 25, wherein the first portion and the second portion are generally planar. [Section 27] 27. The impeller assembly of claim 25 or 26, wherein the at least one blade is at least three blades, such as four blades, five blades, or six blades. [Section 28] 28. The impeller assembly of any one of clauses 25 to 27, wherein the second portion has a radiused distal end. [Section 29] 29. The impeller assembly (880) of any one of paragraphs 25 to 28, wherein the at least one blade (854) further comprises a third portion (882) extending horizontally from the second portion (860). [Section 30] 30. The impeller assembly of clause 29, wherein a distal end (884) of the third portion is wider than a proximal end (886) of the third portion, such that the third portion is generally triangular. [Section 31] 31. The impeller assembly of any one of paragraphs 25 to 30, wherein distal edges (858a, 860a) of the first and second portions are radiused. [Section 32] 32. The impeller assembly of any one of claims 29 to 31, wherein distal edges (858a, 860a, 882a) of the first portion, the second portion, and the third portion are radiused. [Section 33] The hub comprises a generally planar disc (856); 33. The impeller assembly of any one of clauses 25 to 32, wherein the disk comprises at least one slot (872) adjacent each of the at least one blade. [Section 34] 34. The impeller assembly of any one of clauses 25 to 33, wherein the first portion and the second portion form an angle between about 100 degrees and about 180 degrees therebetween. [Section 35] 35. The impeller assembly of claim 34, wherein the first portion and the second portion form an angle therebetween of about 135 degrees, or between 130 degrees and 140 degrees. [Section 36] 36. The impeller assembly of any one of paragraphs 25 to 35, wherein the diameter of the impeller assembly is between approximately one-quarter and about one-half the diameter of the bioreactor (10) in which the impeller assembly is disposed. [Section 37] 37. An impeller assembly as described in any one of clauses 25 to 36, wherein the hub is a magnetic hub. [Section 38] a hub having a central axis; a plurality of blades extending from the hub, at least one of the blades being oriented at one of a lead angle or a retard angle relative to a radial line extending from the central axis of the hub; 1. An impeller assembly for a bioprocessing system comprising: [Section 39] At least one of the plurality of vanes is oriented at a lead angle relative to a first radial line extending from the central axis; 40. The impeller assembly of claim 38, wherein at least one other of the plurality of vanes is oriented at a delay angle relative to a second radial line extending from the central axis. [Section 40] 40. The impeller assembly of claim 39, wherein each of the plurality of blades is oriented at one of a lead angle or a lag angle relative to a respective radial line extending from the central axis of the hub to define a lead blade or a lag blade of the plurality of blades. [Section 41] 41. An impeller assembly as described in any one of paragraphs 38 to 40, wherein the leading vanes and the lagging vanes are positioned alternately in the direction of rotation of the hub. [Section 42] the lead angle is between about 5 degrees and about 30 degrees; 42. The impeller assembly of any one of paragraphs 38 to 41, wherein the delay angle is between about 5 degrees and about 30 degrees. [Section 43] 43. An impeller assembly as claimed in any one of claims 38 to 42, wherein, with respect to the direction of rotation of the impeller assembly, the distance between the tip of a leading blade and the tip of the next adjacent lagging blade is less than the distance between the tip of a lagging blade and the tip of the next adjacent leading blade. [Section 44] 44. The impeller assembly of any one of paragraphs 38 to 43, having a maximum diameter that is between about one-quarter and about one-half the diameter of a bioreactor vessel in which the impeller assembly is configured to be positioned. [Section 45] 1. An impeller assembly for a bioprocessing system, comprising: Hub and a plurality of blades extending from the hub, each having a leading edge and a trailing edge, at least one of the blades comprising an array of slots or openings in a leading edge of the blade; An impeller assembly comprising: [Section 46] 46. ​​The impeller assembly of claim 45, wherein the impeller is a marine impeller. [Section 47] 47. The impeller assembly of claim 45 or 46, wherein the slots are generally vertically extending slots. [Section 48] 48. An impeller assembly as claimed in any one of clauses 45 to 47, wherein each of the blades of the plurality of blades respectively comprises an arrangement of slots or apertures at the leading edge of the blade. [Section 49] 49. The impeller assembly of any one of clauses 45 to 48, wherein the array of slots or openings is positioned at a radial position in the impeller that corresponds to a gas outlet of a sparger assembly of the bioprocessing system. [Section 50] A container (12); a flexible bioprocessing bag (20) positionable within the vessel; 50. The impeller assembly (740; 800; 850; 870; 880; 900; 1000; 1100) of any one of claims 25 to 49, positioned at the bottom of the flexible bioprocess bag; A bioprocessing system (10) comprising: [Section 51] 51. The bioprocess system of claim 50, which is a biological reaction system configured for culturing cells. [Section 52] 52. The bioprocessing system of claim 50 or 51, further comprising a sparger assembly positioned above the impeller assembly. [Section 53] 53. The bioprocessing system of claim 52, wherein the sparger assembly is the sparger assembly (100; 200; 300; 400; 500; 600; 700; 750; 760) of any one of clauses 1 to 15 or 24. [Section 54] 54. The bioprocessing system according to claim 53, wherein the base plate (110; 210; 510; 610; 710; 754) comprises an impeller mounting shaft (124) and a discharge opening (126). [Section 55] 55. The bioprocessing system of claim 53 or 54, wherein the impeller is positioned such that a lower edge of the impeller blades is positioned just above an upper surface of the manifold (112, 124). [Explanation of symbols]

[0078] 10 Biological reaction systems, bioreactors / bioprocess systems 12 vessels, support structure 14 Basics 16 Legs 18 Lifting Assembly 20 Flexible Bags 22 Visual Window 24 Windows 28 Impeller 30 Magnetic Hub 32 Impeller plate 34 Magnetic drive unit, motor 100 Sparger assembly 110 Foundation plate 111 Aeration manifold 112 aeration passage, hollow aeration element, hollow aeration manifold, first aeration manifold 113 Entrance Room Unit 114 Aeration passage, hollow aeration element, hollow aeration manifold, second aeration manifold 115 Entrance room 116 Foot 117 Gasket 118 Support rod, mounting column 119 Perforated plate 120 Gas outlet opening, opening 121 Frames 122 Pipe coupler 123 Cantilever 124 Mounting shaft 125 Protrusion, lip 126 Aperture 127 Auxiliary materials 131 Side wall 133 Bottom wall 200 Sparger assembly 210 Foundation plate 212, 214, 216, 218 Aeration passages, hollow aeration manifolds 220 Foot 222 Mounting column 224 Gas outlet opening, aperture, hole 226 Pipe coupler 228, 230 Piping 232 Mounting shaft 234 Aperture 300 Sparger Assembly 310 T-shaped connector 400 Sparger Assembly 410 Elbow type connector 500 Sparger Assembly 510 Foundation plate 512 Aeration manifold 516 Foot 522 Mounting shaft 600 Sparger Assembly 610 Foundation plate 612, 614 Aeration manifold 616 Gas outlet opening 618 Column section 620 installed shaft 622 Aperture 624 Teeth, top 626 Depression, groove 700 Sparger Assembly 710 Foundation plate 712, 714 Aeration manifold 716 Gas outlet opening 740 Impeller assembly 742 Feather 744 Hub 750 Sparger Assembly 752 Aeration Manifold 754 Foundation plate 760 Sparger Assembly 762 Foundation plate 764 Sparger elements, aeration manifold 766 Gas outlet opening 768 Magnet 770 Magnet 800 impeller assembly 812 Feather 816 First Part 818 Second Part 820 The part where the curve is formed 822 Radius 850 Impeller assembly 852 Hub 854 Feather 856 Distribution disk 858 First Part 858a Distal edge 860 Second Part 860a Distal edge 870 Impeller assembly 872 Slots 882 Third Part 882a Distal edge 884 Distal end 886 Proximal end 900 impeller assembly 910 Hub 912, 914 Feather 916 Direction of rotation 918 Radial Line 920 center axis 1000 impeller assembly 1010 Hub 1012 Feather 1014 Slots 1100 Impeller assembly 1110 Depressions, holes, openings d diameter d1, d2 distance s interval α angle β1 Lead angle β2 Delay angle

Claims

1. A sparger assembly (100; 200; 300; 400; 500; 600; 700; 750; 760) for a bioprocessing system (10), comprising: Base plate (110; 210; 510; 610; 710; 754), at least one aeration manifold (112, 114; 212, 214, 218; 512; 612, 614) removably coupled to said base plate and supported in an elevated position relative to said base plate, each aeration manifold comprising at least one inlet for receiving gas and at least one gas outlet opening (120; 224; 514; 616; 716; 766) for delivering said gas to a fluid in said bioprocessing system; a mounting device (124; 232; 522; 620; 718) that allows for coupling of an impeller (28; 740; 800; 850; 870; 900; 1000; 1100) of the bioprocessing system to the sparger assembly in close association with the aeration manifold; A sparger assembly comprising:

2. 1. An impeller assembly (740; 800; 850; 870; 880; 900; 1000; 1100) for a bioprocessing system, comprising: Hub (30; 810; 852; 910; 1010), at least one blade (742; 812; 854; 912, 914; 1012) operably connected to said hub, said at least one blade having a first portion (816; 858) connected to said hub and extending generally vertically, and a second portion (818; 860) extending at an upward angle from said first portion; An impeller assembly comprising:

3. The impeller assembly of claim 2 , wherein the first portion and the second portion are generally planar.

4. The impeller assembly of claim 2 or 3, wherein the at least one blade is at least three blades, such as four blades, five blades, or six blades.

5. 5. An impeller assembly as claimed in any one of claims 2 to 4, wherein the second portion has a radiused distal end.

6. 6. The impeller assembly of claim 2, wherein the at least one blade (854) further comprises a third portion (882) extending horizontally from the second portion (860).

7. 7. The impeller assembly of claim 6, wherein a distal end (884) of the third portion is wider than a proximal end (886) of the third portion, such that the third portion is generally triangular.

8. 8. The impeller assembly of claim 2, wherein distal edges (858a, 860a) of the first and second portions are radiused.

9. 9. The impeller assembly of claim 6, wherein distal edges (858a, 860a, 882a) of the first, second and third portions are radiused.

10. The hub comprises a generally planar disc (856); 10. The impeller assembly of claim 2, wherein the disk comprises at least one slot (872) adjacent each of the at least one blade.

11. 11. The impeller assembly of claim 2, wherein the first portion and the second portion form an angle between about 100 degrees and about 180 degrees therebetween.

12. 12. The impeller assembly of claim 11, wherein the first portion and the second portion form an angle therebetween of about 135 degrees, or between 130 degrees and 140 degrees.

13. 13. The impeller assembly of any one of claims 2 to 12, wherein the diameter of the impeller assembly is between approximately one-quarter and about one-half the diameter of the bioreactor (10) in which the impeller assembly is disposed.

14. 14. An impeller assembly as claimed in any one of claims 2 to 13, wherein the hub is a magnetic hub.

15. a hub having a central axis; a plurality of blades extending from the hub, at least one of the blades being oriented at one of a lead angle or a retard angle relative to a radial line extending from the central axis of the hub; 1. An impeller assembly for a bioprocessing system comprising:

16. At least one of the plurality of vanes is oriented at a lead angle relative to a first radial line extending from the central axis; The impeller assembly of claim 15 , wherein at least one other of the plurality of vanes is oriented at a delay angle relative to a second radial line extending from the central axis.

17. 17. The impeller assembly of claim 16, wherein each of the plurality of vanes is oriented at one of a lead angle or a lag angle relative to a respective radial line extending from the central axis of the hub to define a leading vane or a lag vane of the plurality of vanes.

18. 18. An impeller assembly as claimed in any one of claims 15 to 17, wherein the leading vanes and the lagging vanes are positioned alternately in the direction of rotation of the hub.

19. the lead angle is between about 5 degrees and about 30 degrees; 19. The impeller assembly of any one of claims 15 to 18, wherein the delay angle is between about 5 degrees and about 30 degrees.

20. 20. An impeller assembly as claimed in any one of claims 15 to 19, wherein, with respect to a direction of rotation of the impeller assembly, the distance between a tip of a leading vane and a tip of a next adjacent lagging vane is less than the distance between a tip of a lagging vane and a tip of a next adjacent leading vane.

21. 21. The impeller assembly of any one of claims 15 to 20, having a maximum diameter of between about one-quarter and about one-half the diameter of a bioreactor vessel in which the impeller assembly is configured to be positioned.

22. 1. An impeller assembly for a bioprocessing system, comprising: Hub and a plurality of blades extending from the hub, each having a leading edge and a trailing edge, at least one of the blades comprising an array of slots or openings in a leading edge of the blade; An impeller assembly comprising:

23. 23. The impeller assembly of claim 22, wherein the blade is a marine blade.

24. 24. An impeller assembly as claimed in claim 22 or 23, wherein the slot is a generally vertically extending slot.

25. 25. The impeller assembly of claim 22, 23 or 24, wherein each of the blades of the plurality of blades is provided with an arrangement of slots or openings, respectively, at the leading edge of the blade.

26. 26. The impeller assembly of any one of claims 22 to 25, wherein the array of slots or openings is located at a radial position in the impeller that corresponds to a gas outlet of a sparger assembly of the bioprocessing system.

27. A container (12); a flexible bioprocessing bag (20) positionable within the vessel; 27. An impeller assembly (740; 800; 850; 870; 880; 900; 1000; 1100) according to any one of claims 2 to 26, positioned at the bottom of the flexible bioprocess bag; A bioprocessing system (10) comprising:

28. 28. The bioprocess system of claim 27, which is a biological reaction system configured for culturing cells.

29. 30. The bioprocessing system of claim 27 or 28, further comprising a sparger assembly positioned above the impeller assembly.

30. 30. The bioprocessing system of claim 29, wherein the sparger assembly is the sparger assembly (100; 200; 300; 400; 500; 600; 700; 750; 760) of claim 1.

31. 31. The bioprocessing system of claim 30, wherein the base plate (110; 210; 510; 610; 710; 754) comprises an impeller mounting shaft (124) and a discharge opening (126).

32. 32. The bioprocessing system of claim 30 or 31, wherein the impeller is positioned such that a lower edge of the impeller blades is located just above a top surface of the manifold (112, 124).

33. A container (12); a flexible bioprocessing bag (20) positionable within the vessel; a sparger assembly (100; 200; 300; 400; 500; 600; 700; 750; 760) positioned at the bottom of the flexible bioprocess bag, the sparger assembly comprising a base plate (110; 210; 510; 610; 710; 754) and at least one aeration manifold (112, 114; 212, 214, 218; 512; 612, 614) removably coupled to and supported by the base plate; A bioprocessing system (10) comprising: each aeration manifold comprising at least one inlet for receiving gas and at least one gas outlet opening (120; 224; 514; 616; 716; 766) for delivering said gas to fluid in said flexible bioprocess bag; 10. The bioprocessing system of claim 1 , wherein the sparger assembly is the sparger assembly of claim 1 .

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