JP2023518693A - Centrifuge system to separate cells in suspension - Google Patents

Abstract

A device for separating a cell suspension into a separation liquid and a concentrate liquid comprises a disposable structure (178, 240, 250) releasably located in a cavity within a solid wall rotatable centrifuge bowl (172). , 370, 414). A portion of the bowl and disposable structure rotates about an axis (174, 428). Stationary inlet feed lines (184, 430), separate liquid discharge lines (212, 436) and concentrate discharge lines (230, 448) extend along the axis of the rotating disposable structure. A separate liquid centripetal pump (208, 438) fluidly connects to the separate liquid discharge line. A concentrate centripetal pump (216, 450) fluidly connects to the concentrate discharge line. At least one concentrate channel (380, 454) and concentrate centripetal chamber (376, 452) are structured to facilitate cell concentrate flow. [Selection drawing] Fig. 1

Description

Field of Invention

The present invention relates to the centrifugation of raw materials. An exemplary configuration of the invention relates to an apparatus for separating cells in suspension by a centrifugation process.

Devices and methods for separating cells in suspension are useful in many technical settings. There is room for improvement in such devices and methods.

Exemplary configurations disclosed herein relate to apparatus and methods for centrifuging cells in large scale cell culture using pre-sterilized, disposable flow path components. The centrifuge used in the present invention can be any solid wall centrifuge that uses pre-sterilized, disposable components and is capable of processing cell suspensions at high cell concentrations.

An exemplary configuration uses a rotatably fixed supply/ejection member. Disposable members often have a flexible membrane attached to a rigid frame having a core with an expanded diameter. The disposable member may further have at least one rescue pump. This disposable structure can be supported in a multi-use rigid bowl with a frusto-conical interior. These structural members allow the system to maintain sufficiently high angular velocities to achieve settling velocities that can efficiently handle highly concentrated cell culture streams. Since it has characteristics that can minimize turbidity at the time of feeding, the cell concentrate can be discharged continuously or semi-continuously, and the overall production rate (production rate) can be reduced to the rate currently used You can do more than that. The structures and methods exemplified herein can be effectively implemented with less risk of contamination.

U.S. Published Application No. 2010/0167388 U.S. Pat. No. 10,384,216 U.S. Patent No. 9,222,067 U.S. Patent No. 6,615,590 U.S. Patent No. 9,427,748

Ramesh S. Gaonker, "Microprocessor Architecture, Programming, and Applications with the 8085" (Prentic Hall, 2002)

FIG. 1 is a schematic diagram showing an exemplary configuration of a centrifuge system with disposable multi-use components. 2 is a detailed view of the upper flange area of the centrifuge system configuration of FIG. 1 showing how the flexible chamber material is sealed to the surface of the flange; FIG. 3 is an isometric cut-away view showing the core and top flange of the disposable member of the centrifuge system configuration shown in FIG. 1; FIG. Figure 4 is a schematic diagram showing the configuration shown in Figure 1, where the pump chamber of the centrifuge system has accelerator fins. 5 is an isometric view of the upper portion of the pump chamber in the centrifuge system configuration shown in FIG. 4; FIG. FIG. 6 is an isometric cut-away view showing the core and upper and lower flanges of a disposable centrifuge system with expanded core diameter (performing shallow pool centrifugation). 7 is an isometric view of the feed accelerator of FIG. 6; FIG. FIG. 8 is an isometric cut-away view showing the core and upper and lower flanges of a disposable centrifuge system with a standard core diameter, a feed accelerator with curved vanes, and an elliptical bowl. 9 is an isometric view of the delivery accelerator of FIG. 8; FIG. FIG. 10 is a schematic diagram showing part of a centrifuge system with continuous discharge of concentrate. FIG. 11 is a schematic diagram showing another configuration having a centrifuge system with continuous discharge of concentrate. FIG. 12 is a schematic diagram showing a centrifuge system with continuous discharge of concentrate with a diluent introduction function. FIG. 13 is a schematic diagram showing yet another configuration of a centrifugal separator system for continuous discharge of concentrate with a centripetal pump throttling mechanism. FIG. 14 is an isometric cut away view showing the core and top flange of a disposable centrifuge system with a core and a feed accelerator with straight blades. 15 is an isometric view of the feed accelerator of FIG. 14; FIG. FIG. 16 is an isometric cut-away view showing another centrifugation system with continuous discharge of concentrate. FIG. 17 is an isometric exploded view showing another centripetal pump. FIG. 18 is an isometric view showing a plate of another centripetal pump with spiral channels therein. FIG. 19 is a schematic diagram showing a centrifuge system operating to ensure that a positive pressure is maintained within the centrifuge core cavity. 20 is a schematic diagram illustrating a simplified exemplary logic flow performed by at least one control circuit shown in FIG. 19; FIG. FIG. 21 is a schematic cross-sectional view of another centrifuge system with continuous centrate and concentrate discharge. FIG. 22 is a schematic cross-sectional view of yet another centrifuge system with continuous separation liquid and concentrate discharge. FIG. 23 is a schematic cross-sectional view of yet another centrifuge system with continuous separation liquid and concentrate discharge. FIG. 24 is a schematic diagram illustrating an exemplary continuous liquid separation and concentrate discharge centrifuge system. 25 is a schematic diagram illustrating the logic flow corresponding to the exemplary control system of FIG. 24; FIG. FIG. 26 is a cross-sectional view showing an exemplary top portion of a disposable centrifuge configuration with a concentrate/separate dam within the separation chamber. FIG. 27 is a cross-sectional view showing an exemplary top portion of a disposable structure having vanes in the separate and concentrate pump chambers to control the radial position of the air/liquid interface. FIG. 28 is a perspective view showing the surface of an exemplary concentrate or separate liquid pump chamber having multiple chamber vanes. Figure 29 is an axial cross-sectional view showing an exemplary top portion of a disposable structure similar to that shown in Figure 27 showing the location of the air/liquid interface. FIG. 30 is an axial cross-sectional view of an exemplary upper portion of a disposable structure with air channels that maintain pressurized air within air pockets. FIG. 31 is a schematic diagram showing an exemplary centrifugation system with backpressure control of the separation liquid flow. FIG. 32 is an axial cross-sectional schematic diagram showing yet another continuous separator/concentrate discharge centrifugation system. 33 is a schematic cross-sectional view of the upper portion of the system shown in FIG. 32; FIG. Figure 34 is a schematic cross-sectional view similar to Figure 32 but showing the system in operation and with an annular cell concentrate region set within the separation chamber. 35 is an external front perspective view of yet another disposable centrifuge structure; FIG. 36 is a cross-sectional view of the disposable structure shown in FIG. 35; FIG. 37 is an exploded view showing the upper disk-shaped portion of the disposable structure shown in FIG. 35; FIG. 38 is a perspective view of the lower portion of the upper disc-shaped portion of the structure shown in FIG. 35; FIG.

In the field of cell culture applied to biopharmaceutical processes, it is necessary to separate cells from a fluid medium, such as the fluid in which the cells are cultured. Target products of cell culture may be molecular species excreted by cells into the medium, molecular species remaining in cells, or cells themselves. At the production (production, production) scale, the early stages of the cell culture process typically occur in bioreactors that can be operated in either batch or continuous mode. Repeated batch processes are also possible. The product of interest often needs to be separated from other process components prior to final purification and product formulation. Cell harvesting is a general term referring to the separation of these cells from other process components. In addition, clarification is a term that refers to cell separation whose purpose is a cell-free supernatant (or separated liquid), and cell recovery is a term that refers to separation whose purpose is a cell concentrate. Exemplary configurations disclosed herein relate to cell harvesting in large scale cell culture systems.

Cell harvest separation methods include batch centrifugation, intermittent centrifugation, continuous centrifugation, semi-continuous centrifugation, tangential filtration (TFF) and depth filtration. Centrifugation for cell harvesting in large-volume cell cultures at production scale has historically been a complex process requiring clean-in-place (CIP) and sterilization-in-place (SIP) techniques to establish an aseptic environment that prevents microbial contamination. It is a very versatile system. Smaller systems are available for laboratory scale and continuous cell harvesting processes. According to the UniFuge centrifuge system, manufactured by Pneumatic Scale Corporation and described in US Published Application No. 2010/0167388, which is fully incorporated herein by reference, in volumes of up to about 2000 liters using intermittent processing Cell harvesting can be successfully performed by batch culture in the range of 3-30 liters/minute. US Pat. No. 10,384,216 and US Pat. No. 9,222,067, both owned by Pneumatic Scale Corporation, the assignee of the present application, are also incorporated herein in their entirety. Generally speaking, intermittent processing requires that both rotation of the centrifuge bowl and feed flow be periodically stopped in order to drain the concentrate. This method of treatment is usually successful in cultures with low concentration and high viability. It should be noted that this method allows large batches to be processed and the cell concentrate to be discharged relatively quickly and completely.

Sometimes it is necessary to harvest cells from high concentration and/or low viability cell cultures that contain high concentrations of cells and cell debris in the feedstock, sometimes referred to as "high turbidity feedstock." Such highly turbid feedstocks can be slow to process in some centrifuge systems. The reason is as follows.
(1) It is necessary to slow down the feed flow rate and increase the residence time in the centrifuge to separate small cell debris particles.
(2) The higher the concentration of both cells and cell debris, the faster the cell concentrate fills the bowl, requiring the bowl to be stopped and the concentrate to drain.
These factors combine to reduce the net processing speed and can lead to unacceptably long cell harvest processing times. In addition to the high costs associated with long processing times, centrifugation times are also long, increasing the risk of product contamination and possible loss of even low viability cell cultures.

High concentrations of cells and cell debris in the feedstock result in very high viscosity cell concentrates. This makes it difficult to completely drain the cell concentrate from the centrifuge, even with extended drain cycles. In some cases, extending the buffer rinse cycle may be sufficient to completely drain the concentrate, but it may be necessary to make either or both of these adjustments to the drain cycle. occurs, and the processing time is long. It becomes more complex and costly to perform large volume cell culture processing.

なお、式中のｖは沈降速度、Δρは固液密度差、ｄは粒子径、ｒは粒子のラジアル位置、ωは角速度、そしてμは液体速度である。スケールアップジオメトリーの場合、ボウルの半径を変えると、粒子が占める最大半径方向位置ｒが変わる。従って、式１の他のパラメーターを一定に維持した場合、ボウル半径が大きくなり、平均沈降速度が増し、所定の分離効率に関して処理量が増える。ところが、半径が長くなると、必要になることもある原料強度が強くなり、また他の工学的な制限もあるため、ボウルの角速度を維持することがより難しくなる。角速度の低下が、比例して大きくなる半径の二乗根を上回ると、平均沈降速度および（半径に比例する）処理量増加が共に減少する。 It is often impractical to scale up the system size by increasing the bowl size and extending the feed portion of the intermittent process cycle. This is because the discharge cycle of the cell concentrate is correspondingly longer. Another limitation is the inability to scale up simple geometries, resulting in variability in the scale up of proper fluid dynamic factors. For any centrifugation, the maximum throughput rate depends on the sedimentation velocity of the particles to be separated. This settling velocity is given by the variation of Stokes' law defined by Equation 1.

Formula 1

In the formula, v is the sedimentation velocity, Δρ is the solid-liquid density difference, d is the particle diameter, r is the radial position of the particle, ω is the angular velocity, and μ is the liquid velocity. For the scaled-up geometry, changing the radius of the bowl changes the maximum radial position r occupied by the particles. Therefore, if the other parameters of Equation 1 are held constant, the bowl radius increases, the average settling velocity increases, and the throughput increases for a given separation efficiency. However, the longer the radius, the higher the raw material strength that may be required, and other engineering limitations, making it more difficult to maintain the angular velocity of the bowl. Both the average settling velocity and the throughput increase (proportional to radius) decrease when the angular velocity drop exceeds the proportionally larger square root of the radius.

One engineering limitation that needs to be considered is that the angular velocities required to rotate a larger bowl are difficult to achieve in practice. This requires a larger, more costly centrifuge drive platform.

Furthermore, if the angular velocity is kept constant while the radius increases, the force pushing the cells towards the wall of the centrifuge also increases. If the bowl is rotated at a sufficiently high angular velocity to achieve the desired treatment efficiency, the stress on the walls of the container and the cells within the container will be high. As for cells, packing them to too high a concentration will damage them. Cell damage is detrimental in applications where cell viability needs to be maintained, leading to contamination of products present in solution in the separation fluid. High viscosities resulting from excessively high cell concentrations can also adversely affect complete evacuation of the cell concentrate.

Exemplary configurations provide continuous or semi-continuous production of low-viability cell suspension cultures containing high concentrations of cells and cell debris at rates suitable for processing large volumes of cell suspension on an industrial scale. Apparatus and methods for continuous centrifugation are included. Some exemplary centrifuges are pre-sterilized, single-use designs and are capable of processing such cell suspensions at flow rates in excess of 20 liters/minute. Because of this flow capacity, the total run time can be set at 2-3 hours for a 2000 liter bioreactor. An exemplary configuration of the single-use centrifuge system can process approximately 300-2,000 liters of fluid while operating at approximately 20-40 liters/minute.

A disposable centrifuge configuration 1000 is shown in FIG. The centrifuge structure 1000 has a core portion 1510, an upper flange 1300, a lower flange 1200, and a flexible liner 1100 sealed to both the upper and lower flanges 1300 and 1200. The core structure 1500 ( 3). The centrifuge structure 1000 comprises a centripetal pump 1400 with a pair of stationary paring discs 1410 in a rotary pump chamber 1420 and a rotary mechanical seal 1700 .

Centrifuge structure 1000 also comprises feed/discharge device 2000 . The device 2000 comprises a plurality of coaxial tubes centered around the axis of rotation 1525 (shown in FIG. 12) of the centrifuge 1000 . The innermost part of the supply/discharge device 2000 comprises supply piping 2100 . A plurality of ancillary lines coaxially surround the supply line 2100 to provide separation liquid discharge 2200, concentrate liquid discharge 2500 (see, for example, FIG. 12), or diluate supply 5000 (see, for example, FIG. 12). Piping or fluid paths may be provided. Each portion of the supply/exhaust connections can be fluidly connected to a portion of the interior of the centrifuge 1000 by appropriate fluid connections, and can also be fluidly connected to coaxial tubing to remove separate, concentrate, or dilute liquid from the system. Fluids can be removed or they can be added to the system.

The upper and lower flanges 1300 , 1200 shown in FIG. 1 are axially aligned with the core portion 1510 and have conical bowls that are concave with respect to the core portion 1510 . The core portion 1510 has a generally cylindrical body with a hollow cylindrical center sized to receive a supply line 2100 having an axis 1525 (best seen in FIG. 12). Top flange 1300, core portion 1510 and bottom flange 1200 can be of unitary construction, providing a more rigid support for flexible liner 1100, sometimes referred to as a membrane. In other configurations, the core portion 1500 can be constructed from multiple members. In other configurations, core portion 1510 and upper flange 1300 may be constructed from a single piece, and lower flange 1200 may be constructed from a separate piece. Alternatively, core portion 1510 and lower flange 1200 may be constructed from a single piece, and upper flange 1300 may be constructed from a separate piece.

An exemplary unitary core portion 1510/upper flange 1300 configuration is shown in FIG. This unitary member can be joined to the lower flange 1200 to form the internal support structure 1500 for the disposable member of the centrifuge 1000 . This structure secures the flexible liner 1100 around a rigid or semi-rigid fixed internal support structure 1500 at the top and bottom. The exterior of the flexible liner 1100 is also supported by the walls of the bowl and cover of the multi-use structure 3000 when the centrifuge system is in use.

The exemplary separation chamber 1550 is a generally cylindrical open chamber that is generally bounded by the outer surface 1515 of the core portion 1510 and the flexible liner 1100 and the upper surface 1210 of the lower flange 1200 and Bounded by lower surface 1310 of upper flange 1300 . Separation chamber 1550 is fluidly connected to supply line 2100 by hole 1530 extending from central cavity 1520 of core portion 1510 to outer surface 1515 of core portion 1510 . This isolation chamber 1550 is also fluidly connected to the pump chamber 1420 by a similar hole 1540 through the core portion 1500 . In this embodiment, hole 1540 is angled upwardly toward pump chamber 1420 and opens into isolation chamber 1550 just below the junction chamber of core portion 1510 and top flange 1300 . As shown in FIG. 12, holes 1420 or 4420 enter the pump chamber at an angle other than an upward angle, such as horizontal or a downward angle. Additionally, in some configurations, slits or gaps between accelerator fins may be used in place of these holes 1420,4420.

Also shown in FIG. 1 is a feed/discharge device 2000 having a feed line carrier 2300 through which the feed line 2100 extends to the position shown in FIG. 3 near the bottom of the centrifuge 1000 . At this position, both the supply function and the discharge function can be exhibited without the supply pipe 2100 moving. Shear forces during the feed process are controlled by careful design of the gap between the nozzle 2110 of the feed pipe 2100 and the upper surface 1210 of the lower flange 1200, and by the diameter and centrifugal force of the nozzle 2110 of the feed pipe 2100. It can be minimized by the angular velocity of the separator. US Pat. No. 6,615,590 (which is incorporated herein by reference) discloses how to select the proper relationship to minimize shear forces. Other suitable supply lines known to those skilled in the art that minimize the shear forces generated in response to the supply of liquid cell culture medium to the rotating centrifuge can also be used.

Also shown in FIG. 1 is a centripetal pump 1400 that discharges the separated liquid via the separated liquid discharge channel 2200 . In the configuration shown in FIG. 1, the separate liquid pump 1400 is located on top of the upper flange 1300 within the pumping chamber 1420 . Pump chamber 1420 is the chamber formed by upper surface 1505 of core 1510 and inner surfaces 1605 , 1620 of centrifuge cover 1600 . Centrifuge cover 1600 may comprise a cylindrical wall portion (shown in FIG. 5) and an engaging cap portion 1610 in the form of a generally circular disc (shown in FIG. 5). The centrifuge cover 1600 may be integrally formed or may be formed from separate members.

In other configurations, the shape and location of the separation liquid pump chamber 1420 may vary, as will be described in more detail below. Pumping chamber 1420 is an axially symmetrical pumping chamber near the top of core structure 1500 , through a hole or slit 1530 extending from near the exterior of core 1515 into separation liquid pumping chamber 1420 . is in fluid connection with isolation chamber 1550 . In some configurations, the separation liquid pump chamber 1420 can be recessed within the separation chamber 1550, as best seen in FIGS.

The exemplary separation liquid pump 1400 includes a pair of pairing discs 1410 . Pairing discs 1410 are two thin circular discs (plates) that are axially aligned with axis 1525 of core structure 1500 . 1-5, the pairing discs 1410 are held stationary against the centrifuge structure 1000 and separated from each other by a constant gap 1415 (1415 in FIG. 15). This gap 1415 between the pairing discs 1410 fluidly connects and removes the separated liquid from the centrifuge 1000 , which flows between the pairing discs 1410 and terminates at the separated liquid outlet 2400 . It flows into a hollow cylindrical separated liquid discharge channel 2200 surrounding 2300 .

Exemplary single-use centrifuge construction 1000 is housed within multi-use centrifuge construction 3000 . This structure 3000 comprises a bowl 3100 and a cover 3200 . The walls of the centrifuge bowl 3100 support the flexible liner 1100 of the centrifuge 1000 as the centrifuge 1000 rotates. Because of this, the outer structure of the single-use centrifuge 1000 and the inner structure of the multi-use structure 3000 match each other. Similarly, the upper surface of the upper flange 1200, the upper outer side of the core portion 1510 and the lower portion of the wall portion 1640 of the centrifuge cover 1600 conform to the inner surface of the multi-use bowl cover 3200 which is adapted to provide rotational support. . The multi-use bowl 1300 and bowl cover 3200 prevent shear forces from tearing the liner 1100 and breaking contact with the single-use centrifuge 1000, as described in more detail below. In some configurations, by selecting a matching disposable centrifuge 1000, an existing multi-use structure 3000 can be incorporated into the disposable process. Also, in other configurations, the multi-use structure 3000 may be specifically designed for use with the single-use centrifuge 1000 .

FIG. 2 shows part of an exemplary construction of top flange 1300, plastic liner 1100 and cover 3200 of multi-use centrifuge structure 3000 to illustrate the sealing of flexible liner 1100 to top flange 1300. show. The flexible liner 1100 can be a thermoplastic elastomer such as polyurethane (TPU) or other tough, tear resistant, biocompatible polymers, and can be formed from top and bottom flanges. 1300 and 1200 may be formed from rigid polymers such as polyetherimide, polycarbonate and polysulfone. Flexible liner 1100 is a thin sleeve or envelope such as that extending between and sealed to upper and lower flanges 1300 , 1200 to form the outer wall of isolation chamber 1550 . Materials for liner 1100 and upper and lower flanges 1300, 1200 and core portion 1510 are exemplary only. A person skilled in the art should be able to select suitable materials with properties similar to known materials.

A thermal adhesive attachment process can be used to bond different materials within the areas shown in FIG. A thermal bond 1110 is formed by preheating the flange material, placing an elastomeric polymer on top of the heated polymer, and applying heat and pressure at a temperature above the softening point of the elastomeric film liner 1100 . Similarly, the plastic liner 1100 is adhered to the bottom flange 1200 . Although thermal bonds 1110 have been described, this is merely exemplary. Other means of establishing an equally strong and relatively permanent bond between the flexible film and the flange material can be substituted, such as temperature, chemicals, adhesives or other bonding means.

An exemplary disposable is pre-sterilized. A heat bond 1110 maintains sterility within the disposable compartment while the components are removed from their protective packaging and installed in the centrifuge. In use, extensible flexible liner 1100 conforms to the walls of reusable bowl 3100 . Because the reusable bowl 3100 provides sufficient support and the flexible liner 1100 exhibits sufficient resilience, the single-use centrifuge 1000 allows large radius centrifuges 1000 to carry liquid cell culture media and other suspensions. It is sufficiently capable of withstanding the strong rotational forces generated when the centrifuge is rotated at angular velocities high enough to reach sedimentation velocities where the turbid liquid is charged and the throughput rate reaches about 2-40 liters/minute.

In addition to the thermal bond 1110, sealing ridges or "nubbins" 3210 are present on the bowl cover 3200 so that the thermoplastic elastomeric film compresses the rigid top flange 1300, further enhancing the sealing action. . The same compression seal is used on the bottom of the bowl 3100 so that the rigid bottom flange 1200 can be sealed with a thermoplastic elastomeric film. These seals support the thermal bond area 1110 by isolating it from shear forces caused by the hydrostatic pressure generated during centrifugation when the chamber is filled with liquid. The seal between the thermal bond area 1110 and the compression nubbin 3210 was tested at 3000 xg, corresponding to a hydrostatic pressure of 97 psi at the bowl wall. The lining should be sufficiently thin and compressible so that the nubins 3210 compress and grip the flexible liner 1100, but with minimal risk of tearing near the thermal bonds 1110 and compression nubs 3210. can be done. In one example configuration, the flexible TPU liner has a seal thickness of 0.010 inches and does not tear or leak.

A configuration corresponding to the illustrations in FIGS. 1 and 2 was tested in a 5.5 inch diameter bowl. The hydraulic capacity at 2000×g was >7 liters/minute and mammalian cells could be successfully separated with 99% efficiency at a flow rate of 3 liters/minute.

In most cases, the top and bottom flanges 1300, 1200 may have a shape similar to that shown in FIG. 1, but in some cases the top surface of the disposable centrifuge may have a different shape, as shown in FIGS. may be 10 and 11, both the upper flange and the bowl cover are disc-shaped when compared to the generally conical bowl cover 3200 which matches the generally conical upper flange 1300. In the configuration shown in FIGS. Those skilled in the art should be able to apply the sealing techniques of the present invention to different shaped sealing surfaces.

4 and 5 illustrate example configurations with features that improve the efficiency of centripetal pump 1400. FIG. As shown in detail in FIG. 5, the internal structure of the disposable member, similar to the internal structure shown in FIGS. Radial fins 1630 may be thin, generally rectangular radial plates that extend perpendicularly from inner surface 1620 of cap portion 1610 . In an exemplary configuration, six fins 1630 are used, but other configurations may have fewer than six fins 1630 or more. In this configuration, the fins form part of the inner surface of cap 1620 , although other configurations may include top surface 1620 of pumping chamber 1420 that may take other shapes than cap 1610 . When the centrifuge system is in use, the fins 1630 rest on the pairing disc 1410 of the centripetal pump 1400 within the pump chamber 1420 . These fins 1630 transmit the angular rotation of centrifuge 1000 to the separated liquid in pump chamber 1420 .

As a result, the efficiency of the centripetal pump 1400 is improved, the gas-liquid interface in the pump chamber 1420 on the pairing disc 1410 is stabilized, and the size of the gas barrier is increased. The gas barrier is generally a cylindrical gas column that extends outwardly from the outside of the feed/discharge mechanism 2000 and into the pump chamber 1420 to the inner surface of the rotating separating liquid. . This increase in the size of the gas barrier is due to the high angular velocity of the separated liquid, which is forced against the walls of the centrifuge. When the spinning separated liquid in the pump chamber 1420 contacts the stationary paring disc 1410, friction can result in reduced efficiency of the pump 1400. FIG. If a plurality of radial fins 1630 that rotate at the same angular velocity as the separation liquid are attached, the decrease in speed can be suppressed. This slowdown results from the collision of the spinning separation liquid with the stationary pairing disc 1410 .

FIG. 6 shows an exemplary configuration showing a core structure 1500 for use with highly turbid feedstocks. Core structure 1500 comprises core portion 1510 , upper flange 1300 and lower flange 1200 . The core portion 1510 has a cylindrical central cavity 1520 into which the supply line 2100 can be inserted. The distance from central axis 1525 to the outside of core portion 1515 (the core portion width indicated by dashed line 6000 in FIG. 6) is longer than the corresponding distance in the configuration shown in FIG. Using a larger diameter core 1510 reduces the depth of separation chamber 1550 (indicated by dashed line 6010), allowing centrifuge 1000 to operate as a shallow pool centrifuge. Generally, the depth 6010 of the isolation chamber 1550 is the distance between the outside of the core portion 1510 shown in FIGS. 1 and 12 and the flexible liner 1100 . For shallow pool centrifuges, the depth 6010 of separation chamber 1550 is small relative to the diameter of the centrifuge. As can be read from the exemplary configuration shown in FIG. 12, shallow pool depth 6010 is shallower at the bottom of separation chamber 1550 and slightly deeper at the top of separation chamber 1550 to facilitate removal of the cell concentrate. Become. In some configurations of the invention, the ratio of average separation pool depth 6010 to core width is 1:1 or less than 1:1. An illustrative example of a shallow pool centrifuge is a selected model of the ViaFuge® centrifuge system manufactured by Pneumatic Scale Corporation. One advantage of shallow pool centrifuges is that they allow separation at higher feed flow rates. For a given bowl inner diameter, this advantage can be realized with a higher average gravity, resulting in a higher settling velocity for a given angular velocity. As a result, the separation properties are improved, which is advantageous when separating highly turbid feedstocks containing high concentrations of cell debris.

The example configuration of core structure 1500 shown in FIG. 6 also has accelerator vanes 1560 as part of lower flange 1200 . Alternative configuration where accelerator vanes 1560 (shown in FIG. 12) fluidly connect the central cavity 1520 of the core 1510 and the separation chamber 1550, rather than holes 1530 drilled into the solid core 1510 (shown in FIGS. 10 and 11). become.

In the exemplary configuration of the core structure 1500 shown in FIG. 6, the accelerator blades 1560 are arranged in a plurality of radial directions (also referred to herein as "radial direction" and "radial", respectively. ), and these vanes extend upwardly from the upper conical surface of the lower flange 1200 . These plates 1580 extend perpendicularly upward to the base of the core portion 1510 . Also, the plate 1580 extends generally radially outward from the vicinity of the axis 1525 of the core portion 1510 . In an exemplary configuration, there are twelve plates 1580, as best illustrated in FIG. In other configurations, the number of plates may be 12 or less, or 12 or more. Furthermore, in other configurations, plate 1580 may be curved in the direction of rotation of centrifuge 1000, as shown in the exemplary configuration of FIG. The inner surface of the lower flange 1200 may be configured to form an elliptical accelerator bowl 1590 from which curved plates extend upward. The above configurations are exemplary, and those skilled in the art will appreciate that they may be combined or configured in different ways to further benefit from the turbidity reduction provided by these plates, the shape of the lower flange 1200, and/or the embedded accelerator. should be able to be partially modified.

Further advantages of the continuous or semi-continuous operation disposable centrifuge 1000 are shown in FIGS. 10-12. The exemplary configuration shown in Figure 10 includes a second centripetal pump 4400 that draws the cell concentrate. Centripetal pump 4400 for removing the cell concentrate is located above centripetal pump 1400 for removing the separation liquid. This centripetal pump 4400 comprises a pumping chamber 4420 and a pairing disc 4410 . A plurality of holes or continuous slits 4540 extend from the upper perimeter of the separation chamber 1550 into the pump chamber 4420 such that the upper outer portion of the separation chamber 1550 is fluidly connected to the second pump chamber 4420 . Similar to pump chamber 1400, pump chamber 4400 may also have a different shape than that shown in FIGS. It has a symmetrical shape. Similar to pumping chamber 1400 , this pumping chamber can be partially or wholly formed by a recess in core structure 1500 . If the separation liquid pump chamber 1400 is near the top of the core structure 1500, the cell concentrate pump chamber 4400 will be provided entirely above it. Pump chamber 4400 fluidly connects with separation chamber 1550 by a hole or slit 4540 to remove the cell concentrate. These slits or the like extend from near the outer top wall of the separation chamber 1550 so that the heavier cell concentrate pushed there by the centrifugal force can be collected.

In the configuration shown in FIG. 10, the pairing disc 4410 used for the concentrate discharge pump 4400 has approximately the same radius as used for the separate liquid discharge pump 1400 and is rotatably fixed. In a configuration such as that shown in FIG. 11, the radius of the pairing disc 4410 of the concentrate exhaust pump 4400 is larger than that of the separate liquid exhaust pump 1400, and the pumping chamber 4420 is correspondingly larger. Pairing discs with various intermediate diameters are also available. The optimum diameter depends on the properties of the cell concentrate to be discharged. The larger the diameter of the paring disc, the higher the pumping capacity, but the greater the shear force generated.

In the configurations shown in Figures 1, 4 and 10, the pairing disc 4410 of the concentrate discharge pump 4400 is rotatably fixed. In other configurations, such as the configuration shown in FIG. 11, the pairing disc 4410 is configured to rotate at an angular velocity between zero and the angular velocity of the centrifuge 1000 . The desired angular velocity can be controlled by a number of mechanisms well known to those skilled in the art. An example of a control means is an external slip clutch that allows the pairing disc 4410 to rotate at an angular velocity that is a fraction of the angular velocity of the centrifuge 1000 . Other means of controlling the angular velocity of the pairing discs will be appreciated by those skilled in the art.

In the configurations shown in Figures 1, 4 and 10-12, the gaps 1415, 4415 between the pairing discs 1410, 4410 are fixed. For configurations such as the configuration shown in FIG. 13, the gap 1415, 4415 between the pairing discs 1410, 4410 can be adjusted to control the flow rates at which separated and concentrated liquids are removed from the centrifuge 1000. FIG. One of each pair of pairing discs 1410 and 4410 is attached to a throttle tube 6100 that can move up and down. The throttle tube 6100 can move up and down to narrow or widen the gap 1415,4415 between each pair of pairing discs 1410,4410. Additionally, an external peristaltic pump 2510 (not shown) can be attached to the concentrate removal line 2500 (not shown) to facilitate removal of the concentrate. This pump 2510 can be controlled by a sensor 4430 in pump chamber 4420 . This sensor 4430 (not shown) can be used to control the diluent pump 5150, thereby allowing concentrate removal to be synchronized with diluent addition.

Also, FIG. 13 shows a configuration in which a separation liquid pump 1400 is provided at the base of the centrifuge 1000 . In the configuration shown in FIG. 13, a separate fluid reservoir 1555 is formed between the pump chamber 1420 and the flexible liner 1100 . A hole 1530 extends from the core portion 1510 underlying the pumping chamber 1420 to this separation fluid reservoir 1555 . Further, in the illustrated exemplary configuration, the separation liquid pump 1400 can be used to remove separation liquid because the hole 1540 extends from the separation chamber 1550 adjacent the outer surface 1515 of the core portion 1510 into the pumping chamber 1420. . In addition, a hole 4540 extends from between the separation chamber 1550 and its upper outer surface into the pump chamber 4420 so that cell concentrate can flow into the pump chamber 4420 and be removed using the centripetal pump 4400 .

As noted above, in the exemplary configuration shown, the gap 1415, 4415 between the pairing discs 1410, 4410 uses a throttle tube 6100 connected to one of each pair of pairing discs 4410, 1410. can be adjusted by The throttle tube 6100 and one of each pairing disc 4410,1410 can be moved up and down to narrow or widen the gap 1415,4415. In the exemplary configuration shown, throttle tube 6100 is attached to the lower and upper pairing discs of pairing disc pair 4410, 1410, respectively. In other configurations, this mounting order may be reversed, one centripetal pump may be throttled, or two centripetal pumps may be arranged in parallel (rather than vice versa as shown in FIG. 13). It is also possible to adjust the aperture.

As can be seen from the configuration shown in FIGS. 10-12, the walls of the solid multi-use bowl 3100 are thicker at the base than at the top, resulting in a frusto-conical interior disposable centrifuge with a smaller radius at the bottom than at the top. A machine structure 1000 can be supported. Thus, due to the larger radius of the upper end of separation chamber 1550 , the more concentrated cell concentrate is directed to the outer upper portion of separation chamber 1550 and into centripetal pump chamber 4420 . In the configuration shown, the multi-use bowl 3100 has a thicker wall at the base than at the top resulting in a frusto-conical shape. Those skilled in the art will recognize that a utility bowl 3100 with a frusto-conical internal shape also has walls of uniform thickness, and that the utility bowl 3100 can be modified to have a purposeful internal shape.

In the exemplary configuration shown in Figures 10-12, the feeding mechanism 2000 also includes additional channels for removing cells or cell concentrates. In the configuration shown in FIG. 1, a cylindrical channel 2200 around a feed line 2100 is used to remove the separation liquid. The configuration shown in FIGS. 10-12 also includes a cylindrical coaxial channel, referred to herein as cell outlet tube 2500, through which cells or cell concentrates are removed. A cell outlet tubing 2500 surrounds the concentrate withdrawal channel 2200 . If the centrifuge is designed for use with concentrates that are expected to be very viscous, an additional cylindrical coaxial channel 5000 can be provided around the feed line 2100 to allow the cell concentrate to A diluent can be introduced into the pump chamber 4420 to reduce the viscosity of the concentrate. In the exemplary configuration shown in FIG. 12, the diluent channel 5000 comprises a coaxial tube that surrounds the cell exit channel, opening at its lower end into a thin disc-shaped channel 5100 on the paring disc 4410 and Fluid communication is provided to the pumping chamber 4420 for evacuation near the outer edge. Injecting the diluent by this means at this location would limit the diluent from being mixed with and discharged with the concentrate rather than being introduced into the separation liquid, depending on the application. Undesirable. Alternatively, the diluent can be introduced directly into the pairing disc and allowed to diffuse radially upwards, or directly into another disc above the pairing disc.

The choice of diluent depends on what the separation process is for and on the nature of the cell concentrate to be diluted. Depending on the case, the diluent may be a simple iso-osmotic buffer or deionized water. Also, it may be advantageous to use a diluent specific to the properties of the cell concentrate. For example, in production-scale batch cell cultures at low cell viability, flocculating agents are commonly added to the medium. This is because cells and cell debris agglomerate into large particles when fed to a centrifuge, and high sedimentation velocities facilitate separation. Since both cells and cell debris have a negative surface charge, compounds used as flocculating agents are generally cationic polymers, such as polyethylenimine, which carry multiple positive charges. Carrying multiple positive charges, such flocculants bind negatively charged cells and cell debris into large aggregates. The use of such flocculating agents can also lead to high viscosity cell concentrates, which is undesirable. Therefore, a particularly useful diluent for the present invention is a deflocculant that interferes with the binding that increases the viscosity of the cell concentrate. An example of a deflocculating agent is a high salt buffer such as sodium chloride solution, the concentration of which is between 0.1M and 1.0M. Other deflocculating agents that are effective in reducing the viscosity of cell concentrates are anionic polymers such as polymers of acrylic acid.

For cell concentrates that are to maintain cell viability, diluents are chosen that are shear protectors such as dextran or Pluronic F-68. Using this shear protector in conjunction with an iso-osmotic buffer increases cell viability upon exit from the centrifuge.

Operation of the exemplary centrifuge shown in FIG. 4 is described below. During the feed cycle, feed suspension enters the rotating bowl apparatus via feed line 2100 . As this feed suspension enters the central cavity 1520 of the core portion 1510 near the lower flange 1200 , centrifugal force forces it outward along the upper surface of the lower flange 1200 and separates it through the holes 1530 in the core portion 1510 . Flow into chamber 1550 .

Separation liquid collects in separation chamber 1550 , the hollow, generally cylindrical space beneath upper flange 1300 surrounding core portion 1510 . Separation liquid flows upward from the inlet until it meets hole 1540 between separation chamber 1550 and pump chamber 1420 at the top of separation chamber 1550 adjacent core portion 1410 and enters the separation chamber via hole 1530. do. Particles denser than the liquid move from the pores 1530 toward the outer wall of the separation chamber 1550 by sedimentation (particle concentrate). When the centrifuge 1000 stops rotating, the particle concentrate moves downward under the influence of gravity into the nozzle 2110 of the supply line 2100 and is discharged by the supply/discharge mechanism 2000 .

During rotation, separation liquid flows through holes 1540 into separation liquid pump chamber 1420 . Within the pumping chamber 1420, the rotating separated liquid impinges on the stationary paring disc 1410, converting the liquid's kinetic energy into pressure that forces it upward through the separated liquid discharge path within the supply/discharge mechanism 2000. It is discharged and exits from the separated liquid discharge pipe 2400 .

The provision of radial fins 1630 on the inner surface 1620 of the cap portion 1610 of the rotating centripetal pump 1400 increases the efficiency of this pump 1400 . These fins 1630 impart the angular momentum of the rotating device to the separation liquid in the pumping chamber 1420 . Otherwise, slowdown may occur due to friction when the rotating separating fluid stationary paring disc 1410 collides. The centripetal pump 1400 provides a better separation liquid evacuation means than mechanical seals due to the air-liquid interface in the pumping chamber 1420 . The gas within the pumping chamber 1420 is not subject to contamination of the external environment by the rotating seal 1700 . Because the separating liquid being drained between the pairing discs 1410 does not come into contact with air during the feeding process, nor during the draining process, excess excess often occurs when air is introduced into the cell culture medium during the draining process. It can prevent foaming.

In the configuration of centrifuge 1000 shown in FIGS. 4 and 5, the cell concentrate periodically interrupts rotation of the bowl and feed flow and then pumps the collected cell concentrate along the outer wall of separation chamber 1550. Ejected by ejecting by. This process is known as intermittent processing. When separation chamber 1550 reaches volumetric capacity, centrifugal rotation is interrupted. The cell concentrate flows down the nozzle 2110 of the supply line 2100 and is collected by pumping it out of the supply line 2100 . A valve (not shown) is operated from the outside of the centrifuge 1000 to collect the concentrated liquid in a collection container (not shown). If the entire batch in the bioreactor has not been completely processed, restart bowl rotation and feed flow and continue the feed/drain cycle until the batch is all processed.

As noted above, the process is slowed when the cell culture medium is concentrated, or if the cell culture medium contains a significant amount of cell debris. This is because the residence time must be extended to capture small debris particles by reducing the feed flow rate, rapidly filling the separation chamber 1550, and frequently and repeatedly interrupting rotation between culture batches. There is a need. In addition, as the cell concentrate becomes more viscous, the action of gravity becomes less effective and the cell concentrate does not reach the bottom of the centrifuge 1000, resulting in the need for extended periods of time and possibly washing. It will be necessary to remove the remaining cells using

A modified single-use centrifuge of the exemplary configuration shown in FIGS. The diluent can be added to the cell concentrate at the time of cell removal, making cell removal easier and more complete.

The construction of the single-use centrifuge structure 1000 shown in FIGS. 6-12 operates as described above. A feed suspension is fed to the disposable centrifuge structure 1000 from a feed tube 2100 . As the feed suspension hits the accelerator vanes 1560 , these vanes 1560 impart an angular velocity to the feed suspension that approximates that of the disposable centrifuge 1000 . The use of vanes 1560 rather than holes 1530 allows a larger volume of feed suspension to be delivered to the separation chamber 1550 at a slower radial velocity, thus providing apertures with a smaller cross-section than the apertures between vanes 1560. It is possible to prevent the jet flow that occurs when feeding the feed suspension into the holes 1530 . This slower feed flow upon entering the separation zone or pool minimizes disturbance of the liquid in the pool and allows sedimentation to be carried out more efficiently.

As the centrifuge 1000 rotates, particles that are denser than the separation liquid are forced out of the separation chamber 1550 , and separation liquid containing no particles approaches the core portion 1510 . The shape of the centrifuge bowl 3100 is an inverted truncated cone, with a radius at the top that is smaller than the radius at the bottom. Centrifugal force causes the particles to collect in the upper, off-center portion (upper outer portion) of the separation chamber. In the case of this centrifuge 1000, an operation of semi-continuously discharging the concentrate is performed. In the case of the discharge of the separating liquid, the operation is carried out as a whole as described with reference to FIG. Evacuation of the cell concentrate is performed in a similar manner, except that cell concentrate that collects near the upper outer portion of the separation chamber 1550 flows into the concentrate discharge pump chamber 4400 through holes 4540 near the upper outer wall of the separation chamber 1550 .

The suspension feed rate and angular rate of rotation are measured using sensors such as, but not limited to, the vibration sensor disclosed in U.S. Pat. can be monitored by With such a sensor system, filling is performed at a lower flow rate until the sensor configuration indicates that the centrifuge is nearly filled, and then the feed flow rate and angular velocity are adjusted appropriately in response to this information. For example, after the centrifuge is almost completely filled, reduce the feed rate or interrupt the feed, increase the angular velocity to increase the sedimentation velocity, and if sedimentation and evacuation are essentially complete, the cycle repeat. If the system is optimized as described above to minimize the need to interrupt the process, it becomes possible to operate the system continuously or nearly continuously at the angular velocity required for sedimentation.

If the concentrate is to be discharged semi-continuously, the suspension is continuously pumped into the centrifuge 1000 using a concentrate pump 4400 that operates intermittently and removes the concentrate. The operation of the concentrate pump 4400 can be controlled by an optical sensor in the concentrate drain line that indicates whether concentrate is being drained. Instead of concentrate pump 4400, the discharge cycle may be managed using controllers and sensors that determine when to open and close valves to most efficiently process the suspension.

Additionally, the average rate of ejection can be controlled by using a centrifuge 1000 with an adjustable gap between the pairing discs 4410,1410. It should be noted that the pair of pairing discs 4410, 1410 need only be adjustable. When the gap between the pairing discs 4410, 1410 (which form part of the flow path exiting the centrifuge 1000) is opened, flow is initiated and closed to stop flow. This gap thus acts as an internal valve. It may be advantageous to widen or narrow the gap 4415, 1415 between the pairing discs 4410, 1410, depending on the product of interest or on the properties of the product. A change in the gap affects both the pumping action and the shear rate associated with the pairing disc.

The rate (amount) of concentrate and segregate removal from centrifuge 1000 and the viability of the removed concentrate can be further controlled using features of the exemplary configurations shown in FIGS. . The addition of accelerator fins 4630 to the concentrate pump chamber 4420, similar to the accelerator fins in the separate pump chamber 1420, eliminates some of the frictional slowdown between the flowing concentrate and the disk 4410, thus reducing the concentration of the concentrate. increase the speed at which the In addition to the accelerator fins 4630 on the top surface of the pump chamber 4420, such fins 4630 can also be attached to the bottom surface of the pump chamber 4420 for added effect. Slits can also be used in place of the holes 1540,4540 to minimize shear forces on the feedstock entering the pumping chambers 1420,4420.

If the viability of the concentrate is a concern, providing a rotatable paring disc 4410 within the pumping chamber 4420 reduces the shear forces applied to the concentrate while in contact with the surface of the paring disc 4410. be able to. Adjusting the rotational speed of the paring disc 4410 to a speed that is between the rest and the rotational speed of the separation chamber 1550 can balance concentrate viability and ejection speed. The desired angular velocity can be controlled by a number of mechanisms known to those skilled in the art. An example of a control means is a slip clutch using which the pairing disc rotates at an angular velocity that is a fraction of the angular velocity of the centrifuge. The use of slip clutches is well known to those skilled in the art. Moreover, there are means other than slip clutches that are obvious to those skilled in the art, all of which can adjust angular velocity.

A peristaltic pump 2510 can also be used to increase the efficiency and reliability of concentrate removal, especially when the feed suspension is highly concentrated. The use of peristaltic pump 2510 allows more precise control of the flow rate of concentrate from centrifuge 1000 than centripetal pump 4400 alone. This is because the centripetal pump speed cannot be adjusted as easily as the peristaltic pump 2510 speed.

Additionally, a diluent pump 5150 can be used to pump a diluent such as sterile water or buffer through the diluent flow path 5000 into the concentrate pump chamber 4420 to reduce the viscosity of the concentrate. See above for a more complete description of useful diluents. The operating speed of either or both centripetal pump 2510 and diluent pump 5150 is controlled by a concentration sensor 4430 responsive to an automatic controller (described below) provided in concentrate outlet connection 2500. can be done. For this controller, pump speeds for both diluent addition and concentrate withdrawal are adjusted according to the concentration of particles in the concentrate, respectively, in response to concentration sensor 4430 and/or according to the standard feed/drain cycle. You can set the program to start, stop, or change the

FIG. 16 shows another exemplary configuration of the core portion used in a centrifuge that continuously feeds the concentrated liquid and the separated liquid and performs continuous separation processing. The core 10 is similar to the cores described above for installation in the rotatable bowl of a centrifuge. During processing, the centrifuge bowl and core rotate about axis 12 . The centrifuge has a stationary device 14 and a rotatable device 16 .

As in the previous embodiment, the stationary device 14 is provided with a supply line 18 . A supply line 18 is coaxial with the axis 12 and terminates in an opening 20 adjacent the bottom of a separate chamber or cavity 22 in the core. The static device further has a separation liquid centripetal pump 24 . Separating liquid centripetal pump 24 , which will be described in detail below, has an inlet opening 26 and an annular outlet opening 28 . The annular outlet opening is provided with a separation liquid line 18 . This annular outlet opening fluidly connects to the separation liquid line 30 . The separation liquid line extends into and coaxially surrounds the supply line 18 .

In this exemplary configuration, the separate liquid centripetal pump 24 is located within the separate liquid pump chamber 32 . The separation liquid pump chamber is part of the rotatable device and is constituted by a wall that becomes the inlet opening 26 of the separation liquid centripetal pump that is exposed to the pool of separation liquid during operation.

Additionally, this exemplary configuration includes a concentrate centripetal pump 34 . Concentrate centripetal pump 34 of this exemplary configuration may also have a configuration similar to that described in detail above. In an exemplary configuration, the concentrate centripetal pump 34 includes an inlet opening 36 located in a wall bounding the annular circumference of the centripetal pump. The outer diameter of the concentrate centripetal pump 34 is larger than the outer diameter of the separated liquid pump. Furthermore, the concentrate pump is provided with an outlet opening 38 . This annular outlet opening 38 is fluidly connected to a concentrate outlet line 40 which extends coaxially around the separate liquid line 30 .

In the exemplary configuration, the concentrate centripetal pump inlet opening 36 is located within the concentrate pump chamber 42 . A concentrate pumping chamber is constituted by a rotatable device 16 . In operation, the inlet opening 36 of the concentrate centripetal pump is exposed to the concentrate within the concentrate pump chamber 42 . Concentrate pump chamber 42 is bounded vertically (up and down) with upper portion 44 . At least one fluid seal 46 extends between the outer periphery of outlet line 40 and upper portion 44 . The exemplary seal 46 is configured to reduce the likelihood of fluid leakage from the interior of the separation chamber and to prevent the introduction of contaminants from the outer regions of the core.

During operation of the centrifuge, the bowl and core with cavity and separation chamber rotate about axis 12 in the direction of rotation. This rotation in the direction of rotation manipulates the cell suspension introduced via feed line 18 into a separate liquid that is discharged via separate liquid line 30 and a concentrate that is discharged via concentrate outlet line 40. Occasionally/behaviorally separate.

The cell suspension enters separation chamber 22 through tubing opening 20 at the bottom of the separation chamber. The cell suspension flows outward due to centrifugal force and multiple accelerator vanes 48 . When the suspension is caused to flow outward by the accelerator vanes, a centrifugal force acts on the cell suspension, causing the cell fluid to flow outward toward the annular tapered wall 50 which bounds the outer separation chamber. As shown, concentrated cell fluid forces outwardly and upwardly against tapered wall 50 and flows through a plurality of concentrate slots 52 . The concentrate flows upwardly over the concentrate slot and into the concentrate pump chamber 42 from which it is expelled by the concentrate centripetal pump 34 .

In the exemplary configuration, in operation, the cell-free separation fluid is located adjacent to the vertical annular wall 54 bounding the interior of the separation chamber 22 . This separation liquid flows upwardly into the separation liquid hole 56 in the annular base structure bounded by the separation liquid pump chamber 32 to form a pool of separation liquid within the separation liquid chamber. From the separation liquid chamber, the separation liquid flows by operation of the separation liquid centripetal pump 24 and is sent from the core portion through the separation liquid piping 30 .

In the exemplary configuration of FIG. 16, the configurations of the concentrate and separate pumps are generally the same as in FIG. In FIG. 17, the separation liquid centripetal pump 24 is shown in isometric view. As shown in FIG. 17, an exemplary separation liquid centripetal pump has a disk-shaped body, which includes first plate 58 and second plate 60 . In operation, the first and second plates are held in releasable engagement by a tightening member such as screw 62 . Of course, other configurations may employ different configurations and fastening methods.

In an exemplary configuration, the second plate 60 comprises walls bounding three sides of a curved spiral channel 64 . It should be noted that in the exemplary configuration, the centripetal pump includes a pair of generally opposed volute channels 64 . Other configurations may employ other numbers and configurations of spiral flow channels.

In an exemplary configuration, the first and second plates comprise a centripetal disk-like body with a vertically extending outer wall 67 forming an annular perimeter 66 . An inlet opening 68 to the spiral channel 64 extends around this annular circumference. An annular collection chamber 70 extends radially outwardly from axis 12 and is fluidly connected to the spiral flow path. An annular collection chamber 70 receives fluid entering from inlet opening 68 . This annular collection chamber 70 fluidly connects to an annular outlet opening coaxial with axis 12 . In this exemplary configuration of the separation liquid centripetal pump, the annular outlet opening is the annular space extending between the outer wall of the supply line 18 and the inner wall of the second plate 60, the outlet of the inner wall of the second plate 60 being It is fluidly connected to the separated liquid outlet line 30 .

In an exemplary configuration, each spiral channel 64 is configured such that the channel curves in the direction of rotation of the bowl and separation chamber. The direction of rotation is indicated by arrow R in FIG. In an exemplary configuration, each of the vertically extending walls 74 bounding the spiral channel and facing in the direction of rotation are curved in the direction of rotation. The curved configuration of the wall 74 horizontally bounding the spiral flow path enhances the pumping characteristics of this exemplary configuration. Additionally, the opposing walls of each spiral flow channel in the exemplary configuration have similar curved configurations. The curved configuration of the vertically extending wall horizontally bordering the spiral channels provides a constant cross-sectional area from the entrance of each spiral channel to the collection chamber. This constant cross-sectional area may also be achieved using a generally flat wall 78 that extends between walls 74, 76 and bounds the spiral flow path horizontally on one side. be. Further, in the exemplary construction, the first plate 58 has a generally planar circular surface 80 on one side which faces inwardly when the plate is constructed into the disk-like body of the centripetal pump. Become. In this exemplary configuration, the circular surface 80 borders both volute 64 sides of the centripetal pump horizontally.

It should be noted that for the above exemplary configuration with a pair of plates, one plate is formed with a recess having walls bounding three of the four sides of the curved spiral flow path. Alternatively, the other plate may form a surface that interfaces with the remaining sides of the spiral flow path, and other configurations may employ other configurations and structures.

The exemplary centripetal pump configuration shown in FIG. 16 provides greater liquid displacement than a similarly sized pairing disk centripetal pump. Additionally, this exemplary configuration requires less liquid heating than a comparable pairing disc.

In the above exemplary configuration, the outer diameter of the annular outer circumference of the separate liquid centripetal pump 24 is smaller than the outer outer diameter of the outer circumference of the concentrate centripetal pump 34 . Using this arrangement, the separation liquid centripetal pump does not draw excessive amounts of liquid from the separation liquid pool formed in the separation liquid pumping chamber 32 . The presence of sufficient liquid in the separate liquid pump chamber prevents waves from forming in the separate liquid adjacent to the inlet of the separate liquid centripetal pump. Wave generation due to insufficient separation liquid not only imparts vibration to the centrifuge and core, but also degrades these other properties.

Due to the large annular perimeter of the concentrate pump of the illustrated configuration, the liquid preferentially flows out of the core by the concentrate centripetal pump. Exemplary configurations allow the downstream flow of concentrate output line 40 to control the ratio of separate to concentrate flow from the core.

In an exemplary configuration utilizing centripetal pumps with the above configurations, furthermore, the characteristics and flow characteristics of the centrifuge can be tailored to the specific raw materials and requirements of the separation process being performed. Specifically, the diameter of the circumference of the centripetal pump can be set to obtain optimum characteristics for a particular process activity. For example, the larger the diameter of the circumference of the centripetal pump, the stronger the flow and the higher the pressure at the outlet. Furthermore, the larger diameters allow for higher mixing ratios than smaller diameters. However, the larger the diameter, the more intense the heating than in the case of a centripetal pump with a smaller peripheral diameter. Thus, a smaller diameter perimeter can be used to keep heating low. It should be noted that the size, area and number of outlet openings may be varied, and different spiral flow path configurations may be employed, in order to tailor flow and pressure characteristics for a particular separation process. be.

FIG. 19 is a schematic diagram of an exemplary system that helps maintain positive pressure within a separation chamber, referred to herein as a cavity, during cell suspension processing. As explained in connection with the above configuration, it is generally desirable to maintain a positive pressure above atmospheric pressure within the separation chamber throughout the process. With this arrangement, there is a risk that contaminants will be introduced into the separation chamber by penetrating one or more fluid seals extending during processing between the stationary device and the core rotatable device. becomes smaller. Further, as previously mentioned, it is generally desirable to maintain a positive pressure of air within the isolation chamber in contact with the inner surface of the fluid seal. The presence of an air pocket adjacent to the seal prevents the seal from coming into contact with the work piece, reducing the risk of introducing contaminants into the treated liquid and of liquid leaking out of the separation chamber.

The exemplary system described with reference to FIG. 19 maintains a constant positive pressure in the separation chamber, thereby reducing the risk of introducing contaminants or leakage of other treated liquids.

The centrifuge comprises a rotatable bowl 82 as shown schematically in FIG. The centrifuge bowl is rotatable about axis 84 by motor 86 or other suitable device.

The illustrated exemplary centrifuge structure includes a rotatable disposable core 88 bounded by a cavity 90, referred to herein as the separation chamber.

As with the other configurations above, this exemplary core includes a stationary device that has a suspension inlet feed line 92 with an inlet opening 94 located adjacent the bottom region of the cavity. Prepare. The stationary device further comprises at least one centripetal pump 96 . A centripetal pump of this exemplary configuration has a disk-like body with at least one pump inlet 98 adjacent its outer periphery and a pump outlet 100 located adjacent the center of the centripetal pump. This pump outlet fluidly connects to the separate liquid outlet line 102 . A separate liquid outlet line extends coaxially around the suspension inlet line in the manner previously described. A rotatable upper portion 104 of the fluid-containing isolation chamber is operatively connected to at least one seal 106 that fluidly seals the core cavity with respect to the inlet and outlet piping. The at least one seal 106 is a rotatable top portion 104 of the core portion having an upper inner wall bounding the stationary separated liquid outlet tubing 102 and, as shown, internally of the cavity 90 . extends in an operationally/operatively sealing relationship between and.

In the exemplary configuration, inlet tubing 92 fluidly connects to pump 108 . Pump 108 in the exemplary configuration is a peristaltic pump useful for non-damaging delivery of the cell suspension. Of course, this type of pump is exemplary only and other types of pumps can be used in other configurations. Further, in the exemplary configuration, the pump 108 is reversible such that the pump 108 operates as a feed pump and pumps a controlled feed of cell suspension from the inlet line 110 into the inlet tubing. In a further exemplary configuration, this pump 108 can operate as a concentrate take-off/drainage pump after separating the cell concentrate by centrifugation. In performing this function, the pump 108 operates to expel the cell concentrate from the separation chamber by reversing the fluid flow in the inlet tubing 92 from the flow that supplied the cell suspension to the separation chamber. The cell concentrate is then discharged into concentrate line 112 . As shown in FIG. 19, inlet line 110 and concentrate line 112 are selectively opened and closed by valves 114 and 116, respectively. In an exemplary configuration, these valves 114 and 116 comprise pinch valves to open and close flow in flexible lines or tubing. Of course, this method is exemplary only, and other configurations may utilize other methods.

In the exemplary system, separated liquid outlet tubing 102 fluidly connects to separated liquid discharge line 118 . This separated liquid discharge line fluidly connects to the separated liquid discharge pump 120 . In an exemplary configuration, separate liquid discharge pump 120 is a variable flow pump that selectively adjusts the flow rate. For example, in some exemplary configurations, pump 120 may comprise a peristaltic pump having a motor whose speed can be controlled to selectively moderate pump flow. The outlet of the separated liquid discharge pump delivers the treated separated liquid to a suitable collection chamber or other processing equipment.

19, the pressure damping reservoir 122 is fluidly connected to the separate liquid outlet line 118 fluidly intermediate the separate liquid outlet piping 102 and the pump 120. As shown in FIG. In an exemplary configuration, the pressure damping reservoir comprises a generally vertically extending vessel to hold the separation liquid in an interior region thereof in liquid-tight relation. The pressure damping reservoir includes a bottom port 124 that fluidly connects to the separate liquid discharge line 118 .

Opposite this reservoir 122 is an upper port 126 . This upper port is exposed to air pressure. In an exemplary configuration, the top port is exposed to air pressure from a high air pressure source, generally indicated at 128 . In an exemplary configuration, the high pressure source comprises a compressor, air storage tank, or other suitable device that provides a source of high air pressure above atmospheric pressure within the range required for operation of the system. Air from the high pressure source 128 is treated with a sterile filter 130 to remove impurities. A regulator 132 maintains a generally constant above-atmospheric air pressure level at the top port of the pressure damping reservoir. In an exemplary configuration, the air pressure regulator includes an electronic fast-acting regulator to ensure that an overall constant air pressure is maintained at a desired level. The exemplary fast regulator 132 activates as soon as the pressure drops below the desired level, increases the pressure acting on the upper port 126, and when the pressure acting on the upper port rises above the regulator setpoint, Immediately release the pressure on the regulator.

In some configurations, the outlet of the regulator may be operatively fluidly connected to the interior of the upper portion 104 of the separation chamber via an air line 143 shown schematically in phantom lines. In such an exemplary configuration, the regulator outlet pressure acting on reservoir top port 126 also acts via air line 143 on an air pocket inside the isolation chamber. This air pocket extends downward to the level of the cavity above the peristaltic pump inlet, into the interior of at least one seal 106, and radially from an area adjacent axis 84 to the upper inner wall inside upper portion 104. Extend. In an exemplary configuration, line 143 applies positive pressure through at least one isolated flow path to a region within the isolation chamber underlying at least one seal. This isolated flow path extends through the stationary structure of the device with the separated liquid outlet line 102 and the inlet feed line 92 . At least one separate passage of air line 143 applies air pressure to the interior of upper portion 104 through at least one air opening that opens into the separation chamber. The at least one opening 145 is located outside the outer surface of the outlet line 102 above the inlet 98 to the centripetal pump and below the at least one seal 106 . Of course, this configuration of air lines to apply positive air pressure inside the isolation chamber air pocket and at least one seal is exemplary only, and other configurations and methods may be used in other configurations.

In an exemplary configuration of pressure damping reservoir 122, upper liquid level sensor 134 detects the separation liquid within the pressure damping reservoir. The upper level sensor operationally/behaviorally detects the separation liquid at the upper level. A low level sensor 136 detects liquid in the reservoir at a lower level. A high level sensor 138 is positioned to detect a high level in the reservoir that is higher than the upper level. This high level sensor detects an unacceptably high level of liquid, indicating an abnormal event requiring system shutdown or other safety precautions. In an exemplary configuration, liquid level sensors 134, 136 and 138 comprise capacitive proximity sensors useful for detecting adjacent separation liquid levels within pressure damping reservoirs. These types of sensors are exemplary only, and other configurations may use other sensors and methods.

Exemplary configurations also include other members suitable for operation of the system. For example, other valves, lines, pressure connections and other suitable components may be provided for proper processing and operation for particular systems such as suspensions, separate liquids and concentrates. For example, an additional valve is valve 140 for controlling the opening and closing of separated liquid discharge line 118 . Additional lines, valves, connections and components thereof that may be used in the system are components that vary depending on the nature of the system.

The exemplary system of Figure 19 includes at least one control circuit 142, which may also be referred to as a controller. The at least one control circuit 142 includes one or more processors 144 . The processor is operationally/operatively connected to one or more data storage devices 146 . A processor, as used herein, means processing data stored in one or more data storage devices or received from an external source, analyzing information, controlling other devices, or otherwise processing data in accordance with processor-executable instructions. means an electronic device that performs the action of One or more control circuits may be implemented as hardware circuits, software, firmware, or applications that enable the control circuit to receive, store, or otherwise process data and perform other actions. For example, the control circuitry may implement one or more microprocessors, CPUs, FPGAs, ASICs, other integrated circuits or other types of circuitry capable of performing functions in the manner of electronic computing devices. Data storage devices include one or more of RAM, flash memory, hard drives, solid state devices, CDs, DVDs, optical memories, magnetic memories, and circuit-readable media for storing computer-executable instructions and/or data. Supports volatile/non-volatile memory.

Circuit-executable instructions include any of a number of programming languages and formats. Examples of these languages and formats include, but are not limited to, routines, subroutines, threads of execution, objects, scripts, methodologies, and functions that perform actions such as those described herein. The structure of the control circuit is described in Ramesh S. et al., incorporated herein by reference. It subsumes, corresponds to, and utilizes the principles described in the textbook entitled "Microprocessor Architecture, Programming, and Applications with the 8085" by Gaonker (Prentic Hall, 2002). . Of course, these circuit structures are exemplary only and other configurations may use other circuit structures for storing, processing, analyzing and outputting information.

In an exemplary configuration, the at least one control circuit 142 is operationally/operatively connected to a suitable interface with at least one sensor, such as sensors 134 , 136 and 138 . At least one control circuit is also operationally/operatively connected to the variable flow displacement pump 120 . Moreover, in some exemplary configurations, at least one control circuit may be operatively/operatively associated with other devices such as motor 86, pump 108, regulator 132, pneumatic pressure source 128, fluid control valves, and other devices. can be connected to

At least one of the control circuits illustrated above operationally/operationally receives data and controls such devices in accordance with circuit-executable instructions stored in data storage device 146 . In an exemplary configuration, the liquid level 147 within the fluid damping reservoir is characteristically corresponding to the pressure within the separation liquid discharge line 102 . One exemplary configuration that does not use the air line 143 utilizes the fact that the pressure in the separation liquid drain line is indicative of the pressure in the upper portion 104 of the core and is indicative of the nature of the pressure in the separation chamber adjacent the seal 106. and controls the operation of the evacuating pump and other components. As previously discussed, it is desirable to maintain a positive pressure above atmospheric pressure and an air pocket adjacent to at least one seal in the separation chamber to prevent introduction of impurities into the separation chamber resulting from negative pressure. It should be noted that if the liquid level becomes too high in the separation chamber, the pressure and treated suspension will overflow the seals, creating potential contamination problems and undesirable exposure and loss of treated liquid. This is because the back pressure on the separate liquid line leading to the outlet from the centripetal pump becomes too high.

In an exemplary configuration, centripetal pump discharge force and pump output pressure level are generated as a function of bowl rotation speed. This pressure output level of the centripetal pump varies with the rotational speed of the bowl and core. In the exemplary configuration without air line 143, back pressure should be controlled at the separate liquid outlet line. Controlling the speed of the motor that operates the pump 120 and the pressure damping reservoir fluid level 147 creates back pressure. The back pressure is maintained below the pump output pressure (so that the centripetal pump can pump the separation liquid out of the separation chamber) and a positive pressure above atmospheric pressure to separate contaminants across the seal. Entry into the chamber is positively prevented and air is maintained at high pressure in the separation chamber adjacent to the seal, separating the seal from the components of the suspension being treated.

In an exemplary configuration, the high pressure applied to the top port 126 of the pressure damping reservoir is maintained by a regulator 132 . In addition, separation flowing out of the separation chamber is controlled by at least one control circuit 142 that maintains the speed of pump 120 and maintains liquid level 147 between upper liquid level 134 and lower liquid level 136 sensed by sensor 134 . The liquid is controlled to maintain the pressure in the upper region of the separation chamber at a desired constant value so that the separation liquid does not contact or overflow the seal.

In an alternative configuration using air line 143, the positive pressure level of the regulator acts on both the fluid in reservoir 122 and the region above the centripetal inlet of the separation chamber. Since the level of positive pressure applied to both of these locations is the same, the back pressure in the separation drain line (the pressure exerted on the fluid in the reservoir) is almost always the same as the pressure in the air pocket at the top of the separation chamber. This allows the centripetal pump to operate completely unaffected by any pressure.

In this exemplary configuration, the pump 120 and other system components are responsive and controllable to at least one control circuit 142 . Therefore, a sufficient volume of air always exists inside the reservoir 122 when the separated liquid is generated. Thus, the reservoir adds a desired damping effect to the pressure change in the separated liquid discharge line to dampen it. This change would otherwise be caused by the pumping action of pump 120 . Calming is accomplished by maintaining the fluid in reservoir 122 below the upper fluid level detected by sensor 134 . In addition, the reservoir fluid level is controlled to remain above the lower fluid level detected by sensor 136 . As a result, the centripetal pump does not discharge air, and the aeration in the separation liquid can be reliably suppressed.

In an exemplary arrangement, the separation liquid flowing out of the separation chamber is controlled by operation of at least one control circuit. An exemplary control circuit operates the system during process conditions to maintain a generally constant flow rate of cell suspension into the separation chamber 90 by the pump 108, while the separation process is controlled by operation of the motor 86. and maintain a constant bowl speed to separate the separation liquid and the cell concentrate. This exemplary configuration also operates to maintain an ideal constant back pressure to the separation liquid discharge line from the centripetal pump while maintaining the air in the separation chamber above the level below the air pocket and at least A single seal 106 separates the separation liquid and the concentrate to be treated.

In an exemplary configuration, the pressure maintained through operation of the regulator in the pressure damping reservoir is set to approximately 2 kpa (0.29 psi) above atmospheric pressure. An exemplary system was found to be set at this pressure to ensure that seal integrity and separability were maintained throughout all stages of cell suspension processing. Of course, this setpoint is exemplary only, and other configurations may employ other pressure setpoints and pressure damping reservoir configurations, sensors, and the like.

FIG. 20 illustrates exemplary logic executed through operation of at least one control circuit 142 in maintaining a desired pressure level in the separation liquid discharge line and within the upper portion of the separation chamber. 1 is a schematic diagram; FIG. It should be noted that control circuitry in some exemplary configurations may perform many additional or different functions beyond those described above. These functions include, in addition to the pressure control functions described above, overall control of the different processes and steps of centrifuge operation. As shown in FIG. 20, in an initial subroutine step 148, at least one control circuit 142 is operated to determine if the centrifuge operation is currently in a mode in which separated liquid is being drained from the separation chamber. can be done. When in this mode, at least one control circuit is activated to operate the separate liquid drain pump 120 to drain separate liquid coming from the separate liquid drain line 118 . To do this, the pump motor should be activated. In an exemplary configuration, the flow rate of pump 120 may be initially set to a set value or may be varied depending on specific operating conditions that may be determined through control circuit operation during the process. Operation of the separate liquid discharge pump is performed at step 150 .

At least one control circuit is then operated to determine in step 152 whether the liquid is at the high level of the high level sensor 138 . If it is at this level, it indicates an undesirable condition. If liquid is detected at the level of sensor 138, the control circuitry is operated to take steps to address this condition. To do this, the pump 120 can be operated to increase the flow rate and, with continued operation of the centrifuge, a determination can be made as to whether the liquid level has fallen within a predetermined period of time. Alternatively, and in addition, the pump 108 can be slowed down by at least one control circuit to reduce incoming liquid flow. If the liquid level does not drop within the predetermined time even after taking such measures, an additional step is taken. In such a process, rotation of bowl 182 may be slowed or stopped. Such measures include deactivating pump 108 to avoid introducing more suspension into the separation chamber. These steps, commonly referred to as normal operation shutdown of the system, can be indicated as step 154 .

If no liquid is detected at the high liquid level sensor 138 , at least one control circuit is operated to determine whether liquid is detected at the upper liquid level of the sensor 134 . This is step 156 . When liquid is detected at the upper fluid level, at least one control circuit operates in response to its stored command to increase the speed of the dispense pump 120 and thus the flow rate. In an exemplary configuration, this is done by increasing the speed of a motor that is part of the pump. This is step 158 . As the pump flow rate increases, the pressure damping reservoir fluid level 147 begins to drop as more fluid is displaced by the pump 120 .

At step 156 , if no liquid is detected at the upper liquid level of sensor 134 , at least one control circuit is operated to determine whether liquid is detected at the upper liquid level of sensor 134 . This is step 160 . If no liquid is detected at the upper fluid level, at least one control circuit operates according to its program to reduce the flow rate of pump 120 . In an exemplary configuration, this is done by slowing down the motor. This is step 162 . In an exemplary configuration, reducing the flow rate of pump 120 causes fluid level 147 to begin to rise within the pressure damping reservoir. In some exemplary arrangements, if the fluid level does not rise within the reservoir within a predetermined time period, the control circuit operates according to its program and takes additional measures, such as stopping step 154 above. The control circuit of the exemplary configuration is activated to vary the pump 120 delivery rate to keep the liquid level 147 in the pressure damping reservoir generally constant between the liquid levels of the sensors 134 and 136 during separation liquid production. maintain a suitable liquid level.

An exemplary configuration maintains a generally constant high pressure of sterile air across the liquid in the pressure damping reservoir, which consistently ensures a similar high pressure in the separation liquid outlet tubing and in the seals within the separation chamber. can be maintained at Exemplary configurations also allow pressure to be maintained at a desired level during different operating conditions of the centrifuge with the bowl rotating at different speeds. These conditions include, for example, when the cell suspension is initially filled into the separation chamber at a relatively high flow rate, and when the centrifuge rotates at a relatively low speed. The pressure can be maintained even during the following conditions for final filling. Under this condition, the flow rate of cell suspension into the separation chamber is lower and the rotation speed of the bowl is higher. In addition, positive pressure can be maintained as previously described during the supply of the suspension to the bowl and the discharge of the separation liquid from the separation chamber. Exemplary configurations also operate at least one control circuit to maintain a positive pressure while the concentrate is expelled from the separation chamber. A positive pressure is maintained in the separation chamber throughout all of these conditions, which reduces the potential for contamination and undesirable conditions. These conditions are otherwise caused by negative pressure conditions (below atmospheric pressure).

Of course, the features, components, structures and control methods described above are exemplary only, and other approaches may be used in other configurations. Furthermore, although the system in the exemplary configuration processes both the separate liquid and the concentrate in batch mode rather than in continuous mode, the principles are applicable to other types of systems.

While the use of a pressure damping reservoir in the exemplary configuration ensures that the desired pressure level is maintained in the outlet line and separation chamber, other methods may be used in other exemplary configurations. For example, in some configurations, pressure may be directly sensed and/or applied to the outlet line, separation chamber, or other location that corresponds to the pressure within the separation chamber. In some configurations, the flow rate of the evacuating pump can be controlled so that a suitable pressure level can be maintained. In yet another configuration, an exemplary control circuit operates to control both the discharge pump and the pump that delivers suspension to the core and/or suitable valves or other flow control devices to provide a suitable pressure level. can also be maintained. The choice of such alternative approach is desirably dependent on the particular centrifuge used and the liquid to be processed.

FIG. 21 is a schematic diagram of another centrifuge system 170 configured for continuous or semi-continuous separation of cell culture batches into a cell separate liquid and a cell concentrate. This exemplary system includes a rigid centrifuge bowl 172 that can rotate about an axis 174 . The bowl includes a cavity 176 configured to releasably receive a disposable structure 178 . This rigid bowl has an upper opening 180 . An annular locking ring or other locking structure 182 releasably secures this disposable structure 178 within the bowl cavity.

An exemplary disposable structure 178 of this configuration includes a central axially extending supply line 184 . This supply line is used to deliver the cell culture batch fluid to the interior region 186 of the disposable structure 178, as described below. A supply line 184 extends from the top at the first axial end 188 of the disposable device to the interior region at the bottom at the second axial end 192 . Disposable structure 178 includes a generally disk-shaped portion 194 adjacent the first axial end. The exemplary disc-shaped portion 194 exhibits a stiffness as a whole. That is, the portion may be rigid or semi-rigid and have an annular perimeter 196 . The annular perimeter is configured to engage the annular boundary wall 198 of the centrifuge bowl cavity 176 . The annular periphery of disc-shaped portion 194 is configured to engage rigid bowl 172 so that the disposable structure rotates therewith.

Additionally, this exemplary disposable structure 178 includes a hollow, rigid or semi-rigid cylindrical core 200 . The core portion 200 is operatively/operatively engaged with the disc-shaped portion 194 and rotates therewith. The core portion 200 is axially aligned with the disk-shaped portion and extends axially halfway between the upper and lower portions of the disposable structure 178 . Core portion 200 includes an upper opening 202 and a lower opening 204 through which supply line 184 passes.

Disc-shaped portion 194 includes a generally circular separation liquid centripetal chamber 206 . A separation liquid centripetal pump 208 is located within the pump chamber 206 . A generally annular separator opening 210 fluidly connects to the separator pump chamber 206 . As used herein, "substantially annular" means discrete and/or continuous apertures in an annular configuration. Separate liquid centripetal pump 208 is fluidly connected to separate liquid discharge line 212 . Separated liquid discharge line 212 extends coaxially in surrounding relationship with supply line 184 . The discharged separated liquid passes through a generally annular opening on the circumference of the centripetal pump and the annular space of the separated liquid discharge line 212 external to the supply line.

Additionally, disc-shaped portion 194 includes concentrate centripetal pump chamber 214 . Concentrate centripetal pump 214 is a generally cylindrical chamber and is located above separate liquid centripetal pump chamber 206 . Concentrate centripetal pump chamber 214 has concentrate centripetal pump 216 located therein. The concentrate centripetal pump is fluidly connected to the concentrate outlet line 220 . Concentrate discharge line 220 extends in annular surrounding relationship with separate liquid discharge line 212 . Concentrate passes through a generally annular opening around the circumference of the concentrate centripetal pump and through an annular space in concentrate discharge line 220 outside of the separate liquid discharge line.

A generally annular concentrate opening 218 fluidly connects to the concentrate pump chamber 214 . In an illustrative example, the generally annular concentrate opening and the generally annular separate opening are concentric coaxial openings, with the concentrate opening radially outward of the separate opening. Of course, this configuration is exemplary only, and other configurations may take other approaches and configurations.

Additionally, the exemplary disposable structure 178 includes a flexible outer wall 222 . This flexible outer wall 222 is a liquid-tight wall and extends in supportive engagement with the wall bounding the rigid bowl cavity 176 in the operative position of the exemplary disposable configuration 178 . In the exemplary configuration, the flexible outer wall 222 operatively engages the disk-shaped portion 194 to provide a rigid fluid connection. The flexible outer wall is internally frusto-conical with a small inner radius adjacent the bottom of the disposable structure adjacent the second axial end 192 .

An exemplary flexible outer wall 222 extends in surrounding relation to at least a portion of the core portion 200 . Additionally, the outer wall 222 bounds an annular separation chamber 224 . Separation chamber 224 extends radially between the outer wall of core portion 200 and flexible outer wall 222 . A generally annular concentrate aperture 218 and a generally annular separate aperture 210 are each in fluid communication with separation chamber 224 .

In an exemplary configuration, flexible outer wall 222 comprises a textured outer surface 226 . This textured outer surface forces air out of the space between the cavity bounding surface of the rigid bowl 172 and the flexible outer wall 222 . In an exemplary configuration, the textured outer surface spans substantially the entire area of the flexible outer wall that contacts the rigid bowl. In an exemplary configuration, the textured outer surface includes outwardly extending protrusions or depressions 228 in one or more patterns to form spaces or depressions therebetween to facilitate ventilation. Venting occurs in the bowl cavity upon placing the disposable structure 178 in the bowl cavity through either the upper opening 180 or the lower opening 230 . In an exemplary arrangement, the protrusions may be constructed from a resilient deformable material and decrease in height in response to the force of the liner against the rigid walls of the bowl. The textured outer surface 226 of the flexible outer wall 222 reduces the risk of air pockets being trapped between the rigid bowl of the centrifuge and the disposable structure. Such air pockets create variations in the wall contours and can lead to unbalanced and/or distorted separation chamber contours that adversely affect the separation process. Of course, the air release structures as described above are merely exemplary, and other configurations may employ other air release structures.

Additionally, the exemplary disposable structure shown in Figure 21 includes a lower disk-like portion 232 that exhibits a rigid or semi-rigid design. This rigid or semi-rigid material allows it to maintain its shape during manipulation. In an exemplary configuration, the lower disc-shaped portion 232 is conical and is operatively/operatively connected to and attached to the lower end of the core portion 200 by a vertically extending wall or other structure. A plurality of angularly spaced passages 234 extend between the upper surface of the disc-shaped portion 232 and the radially outwardly facing lower portion of the core. Channel 232 extends radially outwardly and upwardly relative to the bottom of second axial end 192 so that cells in the cell culture batch entering interior region 186 through opening 190 in feed line 184 are It flows radially outward and upward into the separation chamber 224 .

In the exemplary configuration, the flexible outer wall 222 extends below a lower disc-shaped portion 232 at the second axial end 192 of the disposable structure. The flexible outer wall 222 extends halfway the walls of the rigid bowl 172 bounding the cavity in which the lower disc-shaped portion 232 and the disposable structure are located.

In the exemplary configuration, while centrifuge bowl 172, upper disc-shaped portion 194, lower disc-shaped portion 232 and flexible outer wall 222 are rotating, feed line 184, separate liquid discharge line 212, concentrate discharge line 220, separate centripetal pump 208 and concentrate centripetal pump 216 remain stationary. At least one annular resilient seal 236 extends in operational/operative sealing engagement between the outer surface of the concentrate discharge line 220 and the upper disc-shaped portion 194 . Because at least one seal 236 maintains an airtight seal as described above, an air pocket can be maintained within interior region 186 during cell processing and the seal can be separated from the cell culture batch being processed. An air pocket maintained within the interior region of the disposable structure keeps the separate fluid centripetal pump 208 and the concentrate centripetal pump 216 in fluid communication with the cell culture batch. Similar to the above, a positive pressure can be maintained within the interior region to ensure separation of the at least one seal 236 from the treated cell culture batch. Alternatively, other approaches may be taken to keep the seal separate from the liquid being processed.

Exemplary system 170 operates similarly to that described above. Cells in the cell culture batch flow through supply line 184 into interior region 186 of disposable structure 178 . These cells enter interior region 186 through feed line opening 190 at the lower axial end of the disposable structure. Under centrifugal force, cells flow outward through opening 234 and into separation chamber 224 . The outwardly and upwardly tapered outer wall 222 causes the cell concentrate containing cells or cell sap to collect in the radially outward upper region of the separation chamber 224 . Separation fluid, which is generally free of cells, collects in the separation chamber adjacent radially inwardly to the outer wall of core 200 .

In an exemplary configuration, cell separation fluid flows upwardly through a generally annular separation opening and into a separation fluid pump chamber. This separated liquid passes inwardly through a generally annular opening in the separated liquid centripetal pump and then upwardly through the separated liquid discharge line 212 . At the same time, the cell concentrate flows into concentrate centripetal chamber 214 through generally annular outlet opening 218 . Cell concentrate passes inwardly through a generally annular opening in concentrate centripetal pump 216 before flowing upwardly through concentrate outlet line 220 . This exemplary configuration allows continuous or semi-continuous operation of exemplary system 170 . The operation of the system 170 can be controlled in a manner similar to that described above, so that the system can be operated reliably over long periods of time, with the desired cell concentrate and the total cell-free isolate as separate output fluid streams. can send.

FIG. 22 shows another centrifuge system 238. As shown in FIG. This system 238 comprises a disposable structure 240 . This disposable structure 240 is in many respects the same as the disposable structure 178 previously described. Some of the structures and features of this disposable structure 240 that are generally the same as disposable structure 178 described above are indicated by the same reference numerals as disposable structure 178 .

Disposable structure 240 differs from disposable structure 178 in that it has a lower disc-shaped portion 242 that exhibits a rigid or semi-rigid structure. As for the lower disc-shaped portion 242 , it is of a generally conical structure operationally/operatively connected to the lower end of the core portion 200 . A plurality of radially outwardly and upwardly extending channels 244 extend between the lower end of core portion 200 and lower disc-shaped portion 242 . In addition, the exemplary lower disc-shaped portion 242 includes angularly spaced radially extending vanes 246 . As for the flow passages, they extend radially outward between each pair of angularly adjacent vanes 246 . In this exemplary configuration, the vanes 246 extend upwardly from the bottom of the disc-shaped portion 242, and at least some of the vanes operatively/operatively engage the core at the radially outer portion. In an exemplary configuration, these vanes 246 accelerate cell culture batch processing, facilitating movement and separation within the interior region of the disposable structure.

Another exemplary configuration of centrifuge system 248 is shown in FIG. This exemplary configuration comprises disposable structure 250 . This disposable structure 250 is similar in many respects to the disposable structure 178 described above. Some of the same structures and features of disposable structure 178 described above are indicated by the same reference numerals in disposable structure 250 .

Disposable structure 250 differs from disposable structure 178 in that it includes a lower disc-shaped portion 252 . Lower disc-shaped portion 252 is a generally rigid or semi-rigid conical structure that is operationally/operatively connected to core 200 via a wall or other suitable structure. The lower disc-shaped portion 252 includes a plurality of angularly spaced apart accelerator vanes 254 extending radially outwardly. These accelerator vanes 254 extend downwardly from the lower conical side of disk-shaped portion 252 . Each directly and angularly adjacent pair of vanes 254 has a channel extending therebetween. In this exemplary configuration, flexible outer walls 222 extend intermediate the lower ends of vanes 254 and the walls of rigid bowl 172 bounding cavity 176 . In this exemplary configuration, the accelerator is in the liquid and accelerates the cell culture batch liquid, facilitating the separation occurring within the interior region of the disposable structure. Of course, features of the disposable structures described herein may be combined in different configurations to facilitate separate processing of different types of raw materials and raw materials with different properties and to generate desired output fluid streams. .

FIG. 26 illustrates another disposable structure 304. FIG. The disposable structure 304 is similar to the disposable structure 178 described above, except as noted below. Elements that are the same as disposable structure 178 are shown using the same reference numerals in FIG.

Disposable structure 304 comprises a continuous annular concentrate dam 306 . Concentrate dam 306 extends downward into separation chamber 224 and radially inwardly from generally annular concentrate opening 218 . An exemplary annular concentrate dam, shown in cross-section, has a tapered outer surface that extends downwardly below the concentrate opening and, in axial cross-section, extends outwardly toward opening 218. 308.

Additionally, the disposable structure 304 comprises a continuous annular separation liquid dam 310 . The separated liquid dam 310 extends downwardly in the separation chamber 224 below the generally annular separated liquid opening 210 . The separated liquid dam 310 is located radially outward from the separated liquid opening 210 . In the exemplary configuration, concentrate dam 306 and separate liquid dam 310 extend approximately the same downward distance into separation chamber 224 . It should be noted that other configurations can also be employed in other configuration examples. Also, in other configurations, the centrifuge structure may include either a concentrate dam or a separate liquid dam, but not both.

An annular recess 312 extends radially into the separation chamber between the separate and concentrate dams. This exemplary annular recess extends upward between the separate liquid dam and the concentrate dam to form an annular pocket therebetween.

In the exemplary configuration, the concentrate dam 306 allows primarily cells and other solids to be separated to pass outwardly along the upper portion bounded by the separation chamber 224, opening the concentrate opening 218 and the concentrate. The centripetal pump chamber 214 is reliably reached. Additionally, the separation liquid dam 310 allows the predominantly cell-free separation liquid to flow along the upper surface bounding the separation chamber 224 into the generally annular separation liquid opening 210 and into the separation liquid pumping chamber 206 . becomes possible. It should be noted that many variations can be made to the concentrate dam and the segregate dam in different configurations depending on the nature of the liquid to be treated and the operating conditions of such liquid to be treated.

FIG. 24 is a schematic diagram illustrating an exemplary control system for continuously processing a cell culture as a whole to produce a flow of a generally cell-free separate liquid and a cell concentrate. This exemplary configuration uses the centrifuge system 170 described above. It should be noted that exemplary system features and the like can be used with many different types of feedstocks, centrifuge systems and structures described herein.

In the exemplary configuration shown, centrifuge bowl 172 is rotated by motor 256 about axis 174 at a predetermined speed. Supply tubing 184 is operationally/operatively connected to cell culture supply line 258 that receives cell culture batch fluid. This supply line is operationally/operatively connected to the supply pump 260 . In an exemplary configuration, feed pump 260 may be a peristaltic pump or other pump suitable for delivering cell culture medium at a predetermined flow rate to the disposable structure.

Separated liquid discharge line 212 fluidly connects to separated liquid discharge line 262 . Separate liquid optical density sensor 264 is operationally/operatively connected to the interior region of separate liquid discharge line 262 . In an exemplary configuration, the separation fluid optical density sensor is an optical sensor that operationally/operationally determines the density of cells presently coming from the disposable structure. To do this in an exemplary configuration, an intensity drop in light output may be measured by an emitter received by a receiving device and having at least a portion of the separation liquid stream. The amount of light from the emitter received by the receiving device decreases as the cell density of the separation liquid increases. This is but one example of a sensor that can be utilized to determine the density or quantity of cells present in the separation fluid, and other types of sensors can be employed in other configurations. For example, near-infrared light, other visible light, or invisible light can be used as the light. Other detection configurations may also utilize electromagnetic, acoustic, or other signal forms. The separate liquid discharge line is operationally/operatively connected to the separate liquid pump 266 . In an exemplary configuration, the separation liquid pump may be a peristaltic pump or other variable flow pump suitable for pumping the separation liquid.

In the illustrated configuration, the concentrate drain line 220 is operationally/operatively connected to the interior region of the concentrate drain line 268 . A concentrate optical density sensor 270 is operationally/operatively connected to at least a portion of the interior region of the concentrate discharge line 268 . Exemplary liquid concentrate optical density sensors may be operated in the same manner as the separate liquid optical density sensors described above. It will be appreciated that the concentrate optical density sensor may have different constructions and different characteristics, and that other exemplary configurations may utilize different types of cell density sensors. Concentrate discharge line 268 is operationally/operatively connected to concentrate pump 272 . In an exemplary configuration, concentrate pump 272 may comprise a peristaltic pump or other variable flow pump suitable for pumping concentrate without damage. It should be noted that these structures and components are exemplary only and that other systems may have different or additional components.

The exemplary control system includes control circuitry 274, sometimes referred to herein as a controller. In exemplary implementations, the control circuitry may include one or more processors 276 and may include one or more data storage devices 278 . The one or more data storage devices comprise one or more tangible media, which hold circuit-executable instructions and data, described below when executed by the controller. You can perform operations such as Examples of such media include solid-state memory, magnetic memory, optical memory, and other non-transitory media for holding circuit-executable instructions and/or data. Further, the control circuit may comprise the structures and the like already described.

The operations performed by the exemplary controller 274 will now be described with reference to the logic flow schematic shown in FIG. In the illustrated configuration, the controller 274 operates to control the operation of each component in the system and maintain an output flow that simultaneously generates a cell-free separate liquid and a cell concentrate as a whole. To this end, optical density sensors in the separate and concentrate outlet lines, respectively, are used to detect the cell density (or turbidity) of the output liquid and to adjust the operation of system components to adjust the output. It can be kept within the desired range.

When using the exemplary control system, the concentration of cells in the cell culture to be treated is measured prior to starting the system. Determine the desired axial rotational speed of the centrifuge, as well as the operating speed of the feed pump 260 . In an exemplary configuration, the speed of rotation of the centrifuge and the feed rate of feed cell fluid by the feed pump are generally maintained as constant setpoints by the controller. Of course, other configurations and systems can employ alternative methods in which the controller can regulate the speed and flow rate during cell processing.

In an exemplary configuration, the output rate (flow rate) of the external concentrate pump 272 is set to an initial value (sometimes referred to herein as the "prime value") based on the determined cell concentration. do. The exemplary configuration also sets a "prime duration" that corresponds to the time that the external concentrate pump 272 will initially operate with a default value. The disposable structure 178 is partially filled during this duration. In this exemplary system, the "base rate" is set for the feed rate from feed pump 260 as well as the concentrate pump based on cell density. The base speed of the concentrate pump is the speed (corresponding to the flow rate) at which the concentrate pump will operate after the initial duration. In an exemplary configuration, the set basal time would correspond to a concentrate pump speed that produces a separate liquid with a cell density below the desired set limit and a cell concentrate with a cell density above the desired set range. be done. These setting values and setting ranges are received in response to suitable input device inputs and stored in at least one data storage device.

In the exemplary logic flow shown in FIG. 25, operation of concentrate pump 272 at initial speed is indicated by step 280 . The controller determines whether the concentrate pump is operating at an initial speed for a time corresponding to the initial required time to at least partially fill the disposable structure 178 .

Once the concentrate pump has been running at the initial speed for the initial duration, the controller increases the speed of the concentrate pump to the base speed indicated by step 284 . Controller 274 operates to monitor the cell density of the separation fluid detected by sensor 264 . The controller also determines whether the optical density is higher than the desired set point, as indicated at step 286 . If the optical density of the separate liquid is not above the set point, then the separate liquid is sufficiently depleted of cells and cellular material that the controller does not change the operating speed of the concentrate pump and the logic proceeds to step 284. back to

If at step 286 the optical density of the separation liquid is found to be higher than the set point, then the logic proceeds to step 288 . At step 288, the controller is activated to increase the speed of the concentrate pump by the set incremental step amount. The purpose of this speed increment is to cause the optical density of the separation liquid as a whole to clear the set point as a result of the decrease in the number of cells in the separation liquid.

After increasing the speed of the concentrate pump 272 at step 288, the controller operates in response to sensor 264 to increase the optical density of the separate liquid at step 290 from the speed (flow rate) increment of the concentrate pump for a set period of time. Determine if it is still above the set point after elapse. If so, the controller continues to monitor the optical density of the separate liquid until it is no longer above the set point. In an exemplary configuration, the instructions include set times. This setting is used by the concentrate pump speed controller before the adjustment to the basal speed is found to be sufficient to maintain the optical density of the separate liquid at or below the desired set point. The optical density of the separated liquid should not be higher than the set point over time. In step 292, the increased concentrate pump speed corresponds to a constant production of well-depleted separation liquid or a retention set time separation liquid corresponding to reaching a programmed wait time. is maintained below the set point. In response to a constant production of separation liquid in which the cells have been sufficiently removed, or in response to reaching a programmed wait time, the controller increases the basal speed value of the concentrate pump at step 294 . Make adjustments to correspond to the basal velocity. The controller sets the new base speed and the logic returns to step 284. It should be noted that if step 286 determines that the separate optical density is still above the set point, the concentrate pump speed may be readjusted.

An exemplary controller simultaneously monitors the optical density of cells in the output concentrate stream. To this end, the optical density detected by sensor 270 can be monitored. As shown in step 296, the controller operates to determine if the optical density within the concentrate is below the desired set point. If this optical density is greater than or equal to the desired setpoint value stored in the data storage device, then the cell concentration in the concentrate output stream is greater than or equal to the desired level and the logic returns to step 284 . If the optical density of the concentrate is below the desired set point, which means the cell level within the concentrate is below the desired, the controller moves to step 298 . At step 298, the speed of the concentrate pump is reduced by a predetermined incremental step. As the concentrate pump slows down, the output flow rate decreases and overall the amount of cells in the concentrate output stream decreases, resulting in a higher optical density of the concentrate output stream.

Next, as shown in step 300, concentrate pump 272 operates at the new reduced speed. As shown in step 302, the controller makes a determination as to whether the deceleration is sufficient to operate the concentrate pump at this deceleration rate at a set time corresponding to the setpoint stored in the data storage device. Before, the concentration of cells in the output concentrate stream is high. After this time has elapsed in step 302, the controller returns to step 284 from which the logic flow repeats to determine if further speed adjustments are required.

Of course, this simplified schematic logic flow is exemplary only, and in other configurations, different logic flows and/or additional operating parameters of system components may be used to obtain the desired output flows of separate liquid and concentrate. It may be monitored and adjusted. For example, in another illustrative example, the separation liquid output pump speed, and thus the separation liquid output flow, is at least partially equal to the optical density detected by the separation liquid optical density sensor corresponding to the level of cells in the separation liquid. may be changed in response to For example, if the level of cells in the separated liquid detected exceeds a set limit, the flow rate of the separated liquid may be reduced under the control of the controller. This may be done by a controller, either alternatively or in conjunction with controlling the concentrate discharge flow rate. A controller may alter the separation liquid flow to ensure that the cell level in the separation liquid is maintained below a set limit or within a set range.

Alternatively or additionally, the controller may control the flow rate of the cell suspension into the disposable structure. To do this, the flow rate of the isolate and concentrate from the disposable structure is varied to maintain cell levels in the isolate and concentrate within programmed set limits stored in memory associated with the controller. You may In addition, the controller operates according to its programming to change other process parameters such as bowl rotation speed, diluent introduction, diluent introduction volume, set limits and desired process rate, and Concentrate properties may be maintained. In other exemplary configurations, other properties or parameters can also be monitored or adjusted by the control system to obtain the desired product.

FIG. 27 is a cross-sectional view of yet another disposable centrifuge structure 314. As shown in FIG. This centrifuge structure 314 is generally similar to the disposable separator structure 178 except as follows. This disposable structure 314 includes elements that can operationally/operationally reliably and more stably maintain the air/liquid interface of the air pocket that extends inwardly and separates the seal 236 from the liquid to be treated at the desired radial position. Prepare.

In the disposable configuration 314 , the separation liquid pump 208 is located within the separation liquid pump chamber 316 . The isolate pump chamber 316 is vertically bounded at its bottom by a lower isolate centripetal chamber surface 318 and at its top side by a circular upper isolate centripetal chamber surface 320 .

A lower separate liquid pumping chamber surface 318 extends radially outwardly from the lower separate liquid centripetal pumping chamber opening 322 . In an exemplary configuration, the lower separation liquid centripetal pumping chamber opening 322 extends through the circular top of the core portion 200, and the top opening 202 described above corresponds. A supply line 184 extends through the lower separate liquid centripetal chamber opening.

An upper segregate centripetal chamber surface 320 extends radially outwardly from a circular upper segregate centripetal chamber opening 324 . The supply line 184 and the separated liquid discharge line 212 extend axially through the upper separated liquid centripetal chamber opening.

A plurality of angularly spaced, upwardly extending lower separator chamber vanes 326 extend above lower separator centripetal chamber surface 318 . Lower separate liquid chamber vanes 326 originate from lower separate liquid centripetal chamber opening 322 and extend radially outward. The lower separator vane 326, shown in more detail in FIG. 28, extends radially outwardly from the axis of rotation 174 for a distance V of the lower separator vane. In an exemplary configuration, the lower separate liquid chamber vanes 326 extend upwardly within a circular recess in the lower separate liquid centripetal chamber surface 318 . It should be noted that this configuration is only an example and that other configurations can be used. For example, the radial length of the vanes, the height of the vanes, and the depth and diameter of the recesses can be varied to achieve desired fluid pressure characteristics.

A plurality of angularly spaced downwardly extending upper separator chamber vanes 328 extend from the upper separator centripetal chamber surface 320 . Each of the upper separate liquid chamber vanes 328 originates from the upper separate liquid centripetal chamber opening 324 and extends radially outward. The upper separator vane extends radially outwardly from the axis of rotation 174 the distance it extends the distance of the upper separator vane. In an exemplary configuration, the distance of the upper separating liquid vane corresponds approximately to the distance V of the lower separating liquid vane. In an exemplary configuration, the upper separator vanes extend downward into a circular recess in the upper separator centripetal chamber surface with a similar configuration to the lower separator chamber vanes shown in FIG. 28, but oriented in the opposite direction. is.

In the illustrated exemplary configuration, the separation liquid centripetal pump 208 includes a generally annular separation liquid centripetal opening 330 . A generally annular separation fluid centripetal pump opening 330 extends radially outwardly from the axis of rotation 174 the distance it extends is the distance of the separation fluid pump opening. The separation liquid pump opening distance at which separation liquid centripetal pump opening 330 is located is greater than the distance of the lower separation liquid vane and the distance of the upper separation liquid vane for reasons that will be shown later.

In the exemplary configuration of disposable structure 314 , concentrate centripetal pump 216 is located within concentrate pump chamber 332 . Concentrate pump chamber 332 is vertically bounded on the underside by a circular lower concentrate centripetal pump chamber surface 334 . The separation liquid pump chamber 332 is vertically bounded on the upper side by a circular upper separation liquid centripetal pump chamber surface 336 .

A lower concentrate centripetal chamber surface 334 extends radially outwardly from a lower concentrate centripetal chamber opening 338 . In the exemplary configuration, the lower concentrate centripetal chamber opening corresponds in size to, and is continuous with, the upper isolate centripetal chamber opening 324 . The supply line 184 and the separated liquid discharge line 212 extend through the lower concentrate centripetal chamber opening 338 .

A plurality of angularly spaced, upwardly extending lower concentrate chamber vanes 340 extend over the lower concentrate centripetal chamber surface 334 . Lower concentrate chamber vanes 334 originate from lower concentrate centripetal chamber openings 338 and extend radially outward. The lower concentrate chamber vanes 334 extend radially outwardly from the axis of rotation the distance they extend is the distance of the lower concentrate vanes. In an exemplary configuration, the lower concentrate centripetal chamber vanes 334 extend over circular recesses in the lower concentrate centripetal chamber surface similar to the upper and lower separate chamber vanes. Of course, this configuration is merely an example.

An upper concentrate centripetal chamber surface 336 extends radially outwardly from the upper concentrate centripetal chamber opening 342 . The feed line 184 , the separate liquid discharge line 212 and the concentrate discharge line 220 extend coaxially through the upper concentrate centripetal chamber opening 342 . A plurality of angularly spaced upper concentrate chamber vanes 344 extend downwardly from surface 336 . The upper concentrate chamber vane extends radially outwardly from the upper concentrate centripetal chamber opening 342 the distance it extends the distance of the upper concentrate vane. The upper concentrate chamber vanes extend into upwardly extending circular recesses in the upper concentrate centripetal chamber surface. In an exemplary configuration, the upper concentrate chamber vanes are configured similarly to the lower concentrate chamber vanes and upper and lower separate chamber vanes described above. Of course, this approach is exemplary only and other approaches can be used in other configurations.

Concentrate centripetal pump 216 includes a generally annular concentrate pump opening 346 . The concentrate pump opening is positioned radially from the axis of rotation 174, the distance being the distance of the concentrate pump opening. Of course, this configuration is only an example, and other approaches can be used in other configurations.

In the exemplary disposable structure 314 , the upper and lower concentrate chamber vanes 344 , 340 and upper and lower separate chamber vanes 326 , 328 form an annular air/liquid interface 348 within the separate pump chamber 330 and an annular air/liquid interface 348 within the concentrate pump chamber 332 . Operations are performed to stabilize the air/liquid interface 350 and position them radially. As shown in FIG. 28, the air/liquid interface 348 is located radially midway along the length of the separation chamber vane. That is, it is located radially inward from the separation liquid pump opening 330 . Radially extending separator vanes provide the centrifugal pump force and maintain an annular air/liquid interface 348 in a radial position both above and below the separator centripetal pump. That is, it is located radially inward of the separation liquid pump opening 330 . In the exemplary configuration, the vanes also act to maintain an air/liquid interface, thus maintaining a coaxial circular configuration both above and below the separate liquid pump. Exemplary configurations also allow the radial position of the interface with respect to the axis of rotation to be controlled as described below, so that the separation liquid pump opening 330 can be maintained in the separation liquid at all times and not exposed to air.

Upper concentrate chamber vanes 344 and lower concentrate chamber vanes 340 operate similarly to the separate fluid chamber vanes. These concentrate chamber vanes maintain a circular air/liquid interface 350 within the concentrate pump chamber 332 at a radial distance inside the generally annular concentrate pump opening 346 . This arrangement ensures that the concentrate pump opening is always exposed to concentrate, but not to air. It should be noted that although in the illustrated configuration the separate liquid centripetal pump and the concentrate centripetal pump are approximately the same size, in other configurations the sizes of the centripetal pumps may differ. In such cases, the radial distance from the axis of rotation that the separate chamber vanes and the concentrate chamber vanes extend may differ. The radial position of the air/liquid interfaces in the separate and concentrate pump chambers relative to the axis of rotation may also be different. A number of different vane configurations can be employed depending on the specific relationship between the members comprising the disposable device and the specific workpiece being processed by the disposable structure.

The top portion of yet another disposable structure 352 is shown in FIG. This disposable structure 352 is similar to the disposable structure 304, except as follows. Disposable structure 352 has an air line 354 extending in coaxial relationship surrounding concentrate discharge line 220 . A pneumatic line 354 communicates with an opening 356 inside the disposable structure. These openings 356 extend from the interior of the air line to above the concentrate centripetal pump 216 in the concentrate pump chamber 332 . In this exemplary configuration, the seal 236 operationally/operatively engages the air line 354 and maintains airtight engagement with the air line, as well as the concentrate drain line, the separate liquid discharge line, and the supply line. Additionally, air lines may be utilized to selectively maintain air pressure levels in air pockets within the disposable structure. Such a configuration can utilize the system described above, or an external source of pressurized air, to isolate the seal of the centrifuge structure from the workpiece while maintaining the air/liquid interface at the desired location. It is also applicable to other systems. Of course, this structure is exemplary only, and other configurations may employ other approaches.

FIG. 31 is a schematic diagram showing a system 358 that can be used to continuously separate a cell suspension into a substantially cell-free separate liquid and a concentrate liquid. This system 358 is similar to the system 170 described above, except as follows. In the illustrated configuration, system 358 operates using a disposable structure similar to disposable structure 352 . Controller 274 of system 358 controls the position of the air/liquid interface within the disposable structure, ensuring that the interface is maintained radially inward with respect to the axis of rotation of each of the separate and concentrate pump openings. perform an operation.

In an exemplary configuration, a flow back pressure regulator 360 is fluidly connected to the separate liquid discharge line 262 . In this exemplary configuration, the flow back pressure regulator 360 is fluidly intermediate the separate liquid discharge line 212 and the separate liquid pump 266 . Exemplary system 358 includes pressurized air source 362 . The pressurized air source 362 is connected to a pressure control pilot valve 364 . This control valve is operationally/operatively connected to the controller 274 . A variable pressure in pilot line 366 is selected in response to a signal from controller 274 . Pilot line 366 fluidly connects to back pressure regulator 360 . The pressure applied by the pressure control pilot valve in pilot line 366 can control the separation liquid flow and thus the back pressure of the separation liquid flow applied by flow back pressure controller 360 .

In the exemplary configuration, pressure control valve 368 is in fluid communication with pressurized air source 362 . Control valve 368 is also operatively/operatively connected to controller 274 . In this exemplary configuration, control valve 368 is controlled to selectively apply precise pressure to air line 354 and air pockets within the top of disposable structure 352 .

In an exemplary configuration, controller 274 operates according to stored executable instructions to control operation of system 358 as described with respect to system 170 . The exemplary configuration also operates controller 274 to control pilot pressure valve 364 to vary the back pressure applied by back pressure regulator 360 to separate liquid discharge line 212 . Controller 274 also controls valve 368 . A controller is actuated to maintain and selectively vary the pressure applied to the upper air pocket inside the disposable structure. The controller operates according to its programming to alter the back pressure of the separate liquid flow and/or the air pocket pressure, and to direct the air/liquid interface of the air pocket inward from the separate liquid pump opening 330 and the concentrate pump opening 346. Maintain the rotation axis distance. Pressure changes in both the back pressure of the separate liquid flow and the air pocket, along with the separate liquid chamber vanes and concentrate vanes in this exemplary configuration, maintain air/liquid interface stability and radial outward extent. Therefore, introduction of air from the disposable structure into the separated liquid output and the concentrated liquid output can be reliably suppressed. In addition, the back pressure and flow of the separation liquid can be selected so that the cellular level of the discharged concentrate and correspondingly the detected optical density can be strongly influenced. Thus, the controller operates according to its programming to control concentrate flow rate, segregate backpressure, segregate flow rate, air pocket internal pressure, cell suspension flow rate to the disposable structure, or any other centrifugation process conceivable. Operating variables can be selectively varied to maintain separate and concentrate properties within set limits and/or ranges stored in at least one storage device associated with the controller. Further, the exemplary configuration allows separation and manipulation of different types of raw materials at different flow rates while maintaining reliable control of the separation process. Of course, while control of the position of the air/liquid interface has been described with respect to features of system 170, such control may be employed in other types of systems having other or different types of processing elements. be able to.

Yet another disposable structure 370 is shown in Figures 32-34. This exemplary disposable structure 370 includes various features similar to those described in connection with the other disposable structures 178 , 240 and 250 . It should be noted that additional features used in other disposable structures and described herein can also be used in configurations with the features and relationships shown in disposable structure 370 .

Disposable structure 370 has an upper disc-shaped portion 372 . This exemplary upper disc-shaped portion 372 has a separation fluid centripetal pumping chamber 374 therein. Separate liquid centripetal pump 208 is housed within separate liquid centripetal pump chamber 374 . The separation liquid centripetal pump chamber has a separation liquid chamber volume within an upper disc-shaped portion.

The separation liquid centripetal chamber is in fluid communication with the separation chamber via at least one separation liquid channel 410 . For the separator channel 410 , it is fluidly connected to at least one separator channel inlet 412 . Exemplary at least one separation fluid channel inlet 412 is in fluid communication with the separation chamber proximate to, but extending radially outwardly from, the cylindrical wall of the cylindrical core. In the illustrated exemplary configuration, at least one separator channel inlet 412 is a substantially annular unitary inlet and the separator channel is a substantially annular unitary channel.

Additionally, the upper disk-like portion has a concentrate centripetal pumping chamber 376 . The exemplary concentrate pump chamber 376 is a cylindrical chamber that is horizontally partitioned by a vertically extending circular partition wall 378 . Concentrate centripetal pump chamber 376 has a concentrate chamber volume within an upper disc-shaped portion. In this exemplary configuration, the volume (capacity) of the segregate chamber within the upper disc-shaped portion is greater than the volume of the concentrate chamber for reasons described below.

An exemplary upper disc-shaped portion has an upper piece 394 and a lower piece 396 . In an exemplary configuration, the upper and lower pieces are held in releasable engagement. Of course, this method is exemplary and other configurations may use other methods. In operation or in service, the exemplary lower piece 396 abuts the upper annular interface 398 on its upper side. The exemplary lower piece 396 abuts the lower annular interface 400 on its underside. The upper annular interface 398 has a radially outwardly facing conical annular top surface portion 402 and a radially inwardly facing, radially planar extending top surface portion 404 . The lower annular interface 398 has a radially outward, conical annular lower surface portion 406 and a radially inward, radially planar extending lower surface portion 408 . In the operative position of the upper piece 394 and the lower piece 396, the radially inward, radially planar, horizontally extending lower surface 408 and the radially inward, radially planar, horizontally Extending upper surfaces 404 extend parallel to each other. It should be noted that in the operative position, the conical annular top portion 402 and the conical annular bottom portion need not be in a parallel relationship for reasons explained below.

In the exemplary configuration, a substantially annular cell concentrate channel 380 extends between upper piece 394 and lower piece 396 of upper disc-shaped portion 372 . An annular cell concentrate channel 380 extends radially inwardly from a substantially annular cell concentrate channel inlet 382 . The concentrate channel inlet is located further radially outward from the separate liquid inlet 412 . For the concentrate channel inlet 382, as shown in FIG. 34, a radial It is located in fluid connection with the upper region of the separation chamber 224 at the directional perimeter.

In the exemplary configuration, a substantially annular funnel channel 381 extends upwardly and radially inwardly to an annular cell concentrate channel inlet 382 . An exemplary lower piece 396 of the upper disc-shaped portion 372 in the operative position abuts a substantially planar, radially extending surface 379 on the radially lower inward facing side. The exemplary radially extending surface 379 terminates radially outwardly at the annular edge 377 . In the operational position of the disposable structure, annular funnel channel 381 extends outwardly from annular edge 377 . Additionally, exemplary upper piece 394 of exemplary annular disc-shaped portion 372 has a substantially annular cell concentrate guiding surface 383 . An annular cell concentrate guide surface 383 extends below the annular funnel channel 381 and abuts the separation chamber 224 radially outwardly at the axial level of the radially extending planar surface 379 . In this exemplary configuration, the annular cell concentrate guide surface 383 extends further radially outward and upwardly adjacent the funnel channel.

In this exemplary configuration, annular cell concentrate channel 380 terminates radially inward at concentrate centripetal chamber 376 at a substantially annular cell concentrate outlet 386 . Also, in the exemplary configuration, the annular cell concentrate outlet 386 is positioned to extend to the midpoint of the vertically extending boundary wall 378 . In this exemplary configuration, the concentrate centripetal pump has a substantially annular concentrate centripetal pump inlet 388 . The annular cell concentrate outlet of channel 380 is radially and axially aligned with concentrate centripetal pump inlet 388 .

In the exemplary configuration described above, the annular cell concentrate channel has a tapered portion 390 and a radially extending portion 392 in its axial cross-section. The radially extending portion 392 is directly radially between the radially planarly extending upper surface portion 404 of the lower piece 396 and the radially planarly extending lower surface portion 408 of the upper piece 394 . direction outward. Moreover, the exemplary radially extending portion 392 extends radially outward directly from the annular cell concentrate outlet 386 . In the operational position of the disposable configuration, the horizontally and radially extending portion 392 of the annular cell concentrate channel has a constant channel height. That is, the channel height refers to the dimension of the channel transverse to the direction of concentrate flow in each region of the channel. As a result of this, the horizontally and radially extending portion has a constant cross-sectional area throughout its length. In this exemplary configuration, the horizontally and radially extending portions 392 are axially and radially aligned so that the height in the axial cross-section is level with the inlet of the concentrate centripetal pump.

The tapered portion 390 of the annular cell concentrate channel 380 extends between the conical annular upper surface portion 402 of the lower piece and the conical annular lower surface portion 406 of the upper piece. For the annular cell concentrate channel 380 , the channel height (and channel cross-sectional area) tapers continuously from the cell concentrate channel entrance 382 to the point where the tapered portion fluidly connects to the radially extending portion 392 . configured to have a channel portion that This configuration increases the cross-sectional area of channel portion 390 perpendicular to the direction of concentrate flow and provides greater proximity to the axis of rotation. A continuous gradual increase in the cross-sectional area of the concentrate channel means that the cross-sectional area of the channel at right angles to the direction of concentrate flow increases smoothly in the channel portion, and the channel cross-sectional area changes immediately by 10% or more. It means that there are no discrete steps in locations in the part.

In this configuration, the constant increase in channel height in proximity to the axis of rotation of the disposable system within the tapered portion 390 maintains the desired high fluid velocity of the cell concentrate. In an exemplary configuration, the annular cell concentrate channel avoids areas of pressure drop and thus favors the radially inward velocity of the components of the cell concentrate from the channel inlet 382 to the channel outlet 386. can be maintained.

In the exemplary configuration, annular inlet 382 to annular channel 380 is the portion of the channel with the lowest height and smallest cross-sectional area in the axial cross-section. At the annular inlet 382, cells and the fluid in which they are suspended begin to flow radially inward. In this area of the disposable structure, the cells, which are thicker than the liquid, are subjected to centrifugal acceleration forces directed radially outward by the rotation of the centrifuge. During operation of the exemplary configuration, an external concentrate pump, such as the concentrate pump 272 previously described, is operated to maintain a flow rate that results in an average velocity of the radially inwardly flowing liquid flow at the annular channel inlet 382 . That is, it maintains the sedimentation velocity and the radially outward force acting on the cells caused by the centrifugal force acting on the concentrate at the channel entrance. Further, in some exemplary configurations, the tapered portion 390 has an upwardly tapering configuration such that the sedimentation force component opposing cell flow at the channel entrance and the channel portion is less than the sedimentation force component that opposes cell movement within the radially inward channel.

In an exemplary configuration, the height of the annular tapered portion of the annular concentrate channel increases with decreasing radial distance from the axis of rotation of the disposable structure. In an exemplary configuration of channels in the axial cross-section, the channel height of tapered portion 390, and thus the cross-sectional area perpendicular to the direction of concentrate flow, increases with increasing proximity to the axis of rotation of the disposable structure ( continuously increasing with decreasing radial distance from the axis of rotation). For an exemplary configuration in which the height of the channel increases continuously with decreasing radial distance from the axis, the cells in the cell concentrate will be forced radially inward along with the liquid in which they are suspended. is preferred because it continuously passes through the concentrate channel at . Despite the increased channel height, the cells are subject to a weakened radially outward centrifugal acceleration force with a corresponding decrease in radial distance from the axis of rotation, resulting in a channel entrance 382 A suitable radial inward velocity can be maintained throughout the radially inward migration of the cell concentrate from. Thus, in the exemplary configuration, cell concentrate moves radially inwardly from annular inlet 382 toward cell concentrate outlet 386 and into concentrate centripetal pump chamber 376, and during passage therethrough, cell concentrate. The liquid maintains a suitably high radial inward velocity in channel portion 390 throughout channel 380 .

It should be noted that in the exemplary configuration shown in FIGS. 32-34, the channel portion with continuously increasing cross-sectional area begins at the channel entrance, but in other configurations other approaches and alternative arrangements may be used. can be applied. For example, in some alternative configurations, channel portions having this arrangement may be provided in other locations. Such a position depends on the particular configuration of the concentrate channel and in such a way that the concentrate can achieve sufficiently high flow velocities in the particular portion of the channel to thereby inhibit sedimentation or other forces that impede the desired flow. It can be set according to the conditions necessary for the movement of the concentrate and the cells therein. Further, although this exemplary configuration uses a single annular concentrate channel portion of continuously increasing cross-sectional area, other configurations may use other channels, such as multiple concentrate channels. The configurations shown in FIGS. 32-34 are exemplary only, and other configurations can be used.

In an exemplary configuration, concentrate outlet line 220 fluidly connects to an external concentrate pump during system operation, as previously described. Exemplary configurations may be similar to system 170 previously described, or with other systems responsive to a control circuit that produces an outlet flow of cell concentrate while maintaining a suitable back pressure in the concentrate outlet line. Similarly, the concentrate pump can be activated. It will be appreciated that numerous other components can be implemented in the system that activates the disposable structure 370 to provide the operational capabilities listed herein.

During operation of the exemplary system, disposable structure 370 connects to the centrifuge bowl and separates the cell suspension in the internal region of the structure. The cell suspension is separated into a substantially cell-free cell fraction and into a cell-rich cell concentrate, as previously described. In an exemplary configuration, centrifugation produces an annular cell concentrate region 384 at the radial perimeter of the upper portion of the separation chamber. This exemplary annular cell concentrate region not only maintains fluid connection with the annular channel inlet 382, but also has a substantially annular cell concentrate guiding surface and an annular funnel channel along which the cell concentrates. fluid moves toward channel inlet 382).

In some exemplary configurations, an annular cell concentrate guiding surface 383 bounding the radial perimeter of the separation chamber adjacent to the radially extending surface 379 of the upper disc-shaped portion is actuated to direct the cells. Push the concentrate upwards, pushing it upwards into the annular funnel channel 381 . This is because the guide surfaces extend further radially outward and upwardly close to the final flow path. The centrifugal force generated by the rotation of the centrifuge displaces the cell concentrate forced radially outwardly against the annular cell concentrate guiding surface 383, engaging the guiding surface and flowing into the annular funnel channel. can do. The guide surface guides the cell concentrate, which moves upward and radially inward into the annular cell concentrate inlet. As the cell concentrate is guided radially upward toward the channel inlet 382 by an annular converging surface bounded by a funnel channel of annular axial cross-section, the radial inward velocity of the liquid component of the cell concentrate is increases with decreasing funnel channel area. As previously discussed, the illustrated configuration has the lowest height (and the lowest cross-sectional area), thereby maximizing the fluid velocity of the liquid phase of the cell concentrate at the annular channel inlet 382 .

In an exemplary configuration and its method of operation, the external concentrate pump is actuated and the cell concentrate in the annular cell concentrate channel 380 is forced radially inward of the liquid phase of the cell concentrate, which is higher than the sedimentation velocity of the cells. At a flow rate that produces an average velocity, it produces a flow that moves continuously from the annular cell concentrate inlet 382 to the annular cell concentrate outlet 386 . In an exemplary configuration, a constant high average velocity of the liquid phase of the cell concentrate can be achieved across the height of the channel and over the radial length of the channel, so that the cell concentrate and cells therein form an annular shape. It will favorably flow into the cell concentrate channel. Further, the exemplary configuration includes a continuously increasing cross-section of the tapered portion 390 of the channel, a constant height of the radially extending portion 392, and a generally constant height of the generally annular cell concentrate channel. , because of the relatively small cross-sectional area, there are no regions of significant pressure drop along the length of the channel, and liquid phase velocity and cell continuity can be well maintained throughout the channel.

Moreover, during operation of the exemplary configuration, the cell concentrate entering the concentrate centripetal pump chamber 376 must be sufficiently compressed to ensure that the liquid and cellular phases of the cell concentrate pass radially inwardly through the concentrate centripetal pump inlet. Move at high speed and flow rate. In this exemplary configuration, this result is facilitated not only by the volume and placement of the concentrate centripetal pump chamber, but also by the placement of channel outlet 386 relative to centripetal pump inlet 388 of centripetal pump 216 .

These features of the exemplary configuration facilitate radially inward flow of cell concentrate in the exemplary disposable structure and facilitate operation of the disposable structure and corresponding system. These features and configurations are exemplary, and the principles described herein can be applied to other arrangements and other disposable or multi-use structures to achieve desired performance characteristics and cell separation in other centrifugation processes. Separation can be achieved.

Yet another configuration of disposable structure 414 is shown in FIGS. 35-38. This exemplary alternative disposable structure has many of the features already described and has an upper disc-shaped portion 416 . The outer wall 418 is configured to operatively connect with the centrifuge bowl in which the disposable structure 414 is located. This structure has a lower portion 420 of the outer wall 418 . The disposable structure illustrated in FIG. 36 has an interior region that is frusto-conical in shape and has a smaller inner diameter adjacent the lower portion 420 .

Similar to the previously described configurations, disposable structure 414 has a cylindrical core 422 . The cylindrical core 422 extends axially between the upper and lower portions of the interior region of the structure and has a cylindrical outer partition wall 424 . It should be noted that while cylindrical core 422 is shown in FIG. 36 as a solid structure, in other configurations a hollow core structure can be used. In the exemplary configuration, a cylindrical core 422 extends in an interior region between the bottom of the upper disc-shaped portion and a plurality of upwardly directed, angularly spaced vanes 426 . These vanes 426 have fluid channels between them that extend upwardly from the inside of the wall bounding the lower portion of the interior region of the disposable structure.

In an exemplary configuration, disposable structure 414 is configured to be rotatable about axis 428 within a centrifuge bowl. In addition, the disposable structure has vertically extending supply tubing 430 in the operational position, which tubing receives the cell culture in the interior region of the structure. An exemplary feed line 430 extends downward to a line opening 432 in the lower portion of the disposable structure to the area where the influent to be separated into cell separation and cell concentrate solutions is to be introduced. In the exemplary configuration shown, the supply line 430 extends axially through a cylindrical opening 434 in the core.

In its operational position, the exemplary disposable structure described above has a vertically extending separation liquid drain line 436 . This vertically extending separation liquid discharge line 436 fluidly connects to a separation liquid centripetal pump 438 . A separation liquid centripetal pump is located within a separation liquid centripetal chamber 440 located within the upper disk-shaped portion 416 . Separation liquid centripetal chamber 440 is fluidly connected to separation chamber 442, which extends radially between the outer wall of core 422 and the wall that abuts the inner region of the disposable structure. The separation liquid centripetal chamber fluidly connects to the separation chamber 442 via at least one separation liquid channel inlet 444 and at least one separation liquid channel 446 . In an exemplary configuration, the at least one separation fluid channel inlet 444 is located in the separation chamber radially outward from, but radially adjacent to, the cylindrical wall bounding the core 422 . In an exemplary configuration, the shape of the separation liquid channel inlet is arcuate. Further, in the operative position of the exemplary configuration, the separation liquid centripetal pumping chamber 440 has horizontally extending upper and lower separation liquid pumping chamber surfaces which are aligned with the upper and lower separation liquid chamber surfaces, such as the vanes 326 and 328 previously described. It can have wings.

Additionally, the exemplary disposable structure 414 has a concentrate drain line 448 that extends perpendicular to the operational position. As with the other configurations described above, concentrate drain line 448, separate liquid drain line 436 and feed line 430 are coaxially arranged on the disposable structure. Concentrate outlet line 448 fluidly connects to concentrate centripetal pump 450 . Concentrate centripetal pump 450 is located within concentrate centripetal chamber 452 within upper disc-shaped portion 416 . In an exemplary configuration, each concentrate centripetal chamber is bounded by an upper and lower concentrate centripetal chamber surface, each surface can have radially extending chamber vanes similar to the vanes previously described.

In this exemplary configuration, the concentrate centripetal chamber fluidly connects to the separation chamber via a plurality of radially extending concentrate channels 454 . In an exemplary configuration, each concentrate channel extends into the upper disc-shaped portion and is angularly spaced from each other channel. The exemplary upper disc-shaped portion 416 consists of an upper piece 456 and a lower piece 458 . Additionally, the exemplary top disc-shaped portion has a bottom piece 460 . In the operational exemplary configuration, top piece 456, bottom piece 458 and bottom piece 460 are each in sandwich-like engagement. In the operational position, each of the plurality of concentrate channels abuts at least one downwardly facing surface of upper piece 456 and at least one upwardly facing surface of lower piece 458 .

As shown in FIG. 36, each of the plurality of concentrate channels 454 has a concentrate channel inlet 462 . In an exemplary configuration, the concentrate channel inlets have a minimum cross-sectional area in a direction perpendicular to the direction of concentrate flow throughout each respective concentrate channel. Each concentrate channel inlet 462 fluidly connects to the perimeter of the separation chamber via a vertical opening 464 extending into the upper disc-shaped portion. In the exemplary configuration, each concentrate channel 454 passes through the bottom piece 460 and is fluidly connected to the separation chamber via a respective vertical opening 464 between and bordering the top piece 456 and the bottom piece 458 . do. In the exemplary configuration, each vertical opening 464 has an arcuate elongated slot in the bottom piece 460 . It will be appreciated that this arrangement is exemplary and other approaches may be used in other configurations.

In the exemplary configuration, each concentrate channel has a generally constant cross-sectional width from the channel inlet 462 to each opening from the channel to the concentrate centripetal chamber. Each concentrate channel has channel portions that continuously increase in cross-sectional area perpendicular to the direction of concentrate flow within each concentrate channel portion. This cross-sectional area increases as the location of the channel portion increases in proximity to the axis of the disposable structure. In an exemplary configuration, for multiple concentrate channels, the cross-sectional area perpendicular to the direction of concentrate flow in tapered portion 468 continuously tapers as a result of varying channel heights in the tapered portion. configured to The height of each channel increases with increasing proximity to the axis of rotation. Of course, this configuration is exemplary and other configurations may take other approaches.

In the exemplary configuration, each channel portion of continuously increasing cross-sectional area arrangement begins at a respective concentrate channel inlet 462 and continues through an upwardly and radially inwardly extending tapered portion 468 . Each tapered portion 468 fluidly connects to a horizontally and radially extending portion 470 of a respective concentrate channel 454 . In the exemplary configuration, each horizontally and radially extending portion of each concentrate channel extends from tapered portion 468 to concentrate centripetal chamber 450 . In an exemplary configuration, the horizontally and radially extending portion of the concentrate channel is of constant cross-sectional area in a direction perpendicular to concentrate flow radially inward from the tapered portion to the concentrate centripetal chamber. be. It should be noted that this configuration is exemplary and other approaches may be applied in other configurations.

During operation of the exemplary disposable structure 414, the upper disc-shaped portion 416, wall 418 and core pivot and connect to the centrifuge bowl. The supply line 430 , the concentrate drain line 436 and the concentrate drain line 448 are stationary along with the concentrate centripetal pump 450 and the concentrate centripetal pump 438 . In an exemplary configuration, the cell culture is separated into cell separate liquid and cell concentrate in separation chamber 442 by rotationally induced centrifugal force. The cell concentrate accumulates in the upper and radially outer region of the separation chamber 442, while the substantially cell-free separation liquid accumulates in the separation chamber region adjacent the cylindrical outer wall of the core.

Separation liquid enters the separation liquid centripetal chamber via a plurality of separation liquid channel inlets 444 and exits the separation liquid centripetal chamber via a separation liquid centripetal pump 438 and a separation liquid exhaust line. An external concentrate pump, such as the concentrate pump already described, is operatively connected to the concentrate outlet line so that the concentrate flows out of the disposable separation chamber. This concentrate flow causes cell concentrate to move upward through vertical openings 464 and through concentrate inlets 462 where the relatively small cross-sectional area of the concentrate channel inlets accelerates the cell concentrate. . The high fluid and cell flow rates in the cell concentrate exert a force on the cells contained in the cell concentrate to overcome the radially outward force acting on the cells as described in relation to the conventional configuration. causes the cells to enter the upward and radially inward tapered portion 468 .

The upwardly and radially inwardly extending tapered portion continuously tapers in cross-sectional area perpendicular to the direction of concentrate flow. This continuously increasing cross-sectional area allows the flow rate of the concentrate to be maintained sufficiently high at each radial location throughout the channel portion so that the cells are displaced despite the force acting radially outward at each location of the channel portion. The concentrate will flow into the concentrate chamber at a favorable high velocity. In an exemplary configuration, the cell concentrate flows out the tapered portion of the concentrate channel and through the horizontally and radially extending portion of each channel into a concentrate centripetal pump chamber that maintains a suitable high velocity of the cell concentrate. Reach 452. As a result, the exemplary configuration provides the cell concentrate with flow characteristics that facilitate cell concentrate flow within the disposable structure and facilitates the cell separation process. Of course, the configuration of disposable structure 414 is exemplary, and other configurations may utilize other configurations.

In some exemplary configurations, the top, bottom and bottom pieces of upper disc-shaped portion 416 can be releasably engaged. This facilitates manufacturing and allows inspection, cleaning, etc. In other exemplary configurations, the pieces can remain in permanent engagement. In yet another exemplary configuration, for structures similar to those described herein, for structures similar to those described herein, other It can be formed from constituent components. Additionally, the configuration arrangement of the disposable structure 414 is exemplary and the useful principles and structures described herein can be used with other separator structural configurations.

Thus, the novel centrifuge system and method of the illustratively disclosed configuration achieves at least some of the objects set forth above and overcomes the problems encountered when using conventional devices and systems. can be solved, and the desired result described above can be obtained.

Certain terminology has been used in the above description for the sake of brevity, clarity, and understanding, and should not be construed as unnecessary limitation. These terms are intended to be descriptive and are intended to be broadly interpreted. Furthermore, the description and explanations herein are exemplary, and the invention is not limited to the exact details shown and described.

It should be noted that features and/or relationships associated with one example configuration can be combined with features and/or relationships in another example configuration. That is, various features and/or relationships can be combined in further configurations. The inventive scope of this disclosure is not limited to only the exemplary configurations shown herein and illustrated in the drawings.

In the claims, any feature recited as a means of performing a function is to be construed to encompass any means known by those of ordinary skill in the art to perform that function, and is not intended to be used herein. It is not intended to be limiting to the structure shown, or the mere equivalent thereof.

Having described the discovery of new and useful features and principles, how to make, use and operate them, and the advantages obtained, new and useful structures, devices, elements, arrangements, parts, combinations and systems. , acts, methods and relationships are as set forth in the claims.

10 Core 12 Axis 14 Static device 16 Device 18 Supply pipe 20 Opening 22 Cavity 24 Separating liquid centripetal pump 26 Inlet opening 28 Outlet opening 30 Separating liquid piping 32 Separating liquid pump chamber 34 Concentrate centripetal pump 36 Inlet opening 38 Outlet opening 40 Concentrate outlet pipe 42 Concentrate pump chamber 44 Upper part 46 Fluid seal 48 Accelerator vane 50 Tapered wall 52 Concentrate slot 54 Annular wall 56 Separating liquid hole 58 First plate 60 Second plate 62 Screw 64 Spiral channel 66 Perimeter 67 Outer wall 68 Inlet opening 70 Annular collection chamber 74 Wall 76 Wall 78 Wall 80 Circular surface 82 Bowl 84 Axis 86 Motor 88 Core 90 Cavity/separation chamber 92 Suspension inlet feed line 94 Inlet opening 96 Centripetal pump 98 Inlet 100 Pump outlet 102 isolate outlet tubing 104 top 106 seal 108 pump 110 inlet line 112 concentrate lines 114, 116 valve 118 isolate drain line 120 isolate drain pump 122 pressure damping reservoir 124 bottom port 126 top port 128 top port 130 sterile filter 132 Controllers 134, 136 and 138 Level Sensor 140 Valve 142 Control Circuit 143 Air Line 144 Processor 145 Aperture 146 Data Storage Device 147 Level 148 Subroutine Step 150 Step 152 Step 154 Step 156 Step 158 Step 160 Step 162 Step 170 Centrifuge System 172 Centrifuge Bowl 174 Axis 176 Cavity 178 Disposable Structure 180 Top Opening 182 Fixing Structure 184 Supply Line 186 Interior Region 188 First Shaft End 190 Opening 192 Second Shaft End 194 Disc Shaped Portion 196 Annular Perimeter 198 Annular Boundary Wall 200 Core Part 202 Upper opening 204 Lower opening 206 Separation liquid centripetal pump chamber 208 Separation liquid centripetal pump 210 Separation liquid opening 212 Separation liquid discharge pipe 214 Concentrate liquid centripetal pump chamber 216 Concentrate liquid centripetal pump 218 Concentrate liquid opening 220 Concentrate liquid discharge pipe 222 Outer wall 224 Separation Chamber 226 Textured Outer Surface 228 Indentation 230 Lower Opening 232 Disc-Shaped Portion 234 Channel 236 Seal 238 Centrifuge System 240 Disposable Structure 242 Lower Disc-Shaped Portion 244 Channel 246 Vanes 248 Centrifuge System 250 Disposable Structure 252 Bottom Disk-like portion 254 Accelerator vane 256 Motor 258 Cell culture feed line 260 Feed pump 262 Separate liquid discharge line 264 Separate liquid optical density sensor 266 Separate liquid pump 268 Concentrate discharge line 270 Concentrate optical density sensor 272 Concentrate pump 274 Control circuit 276 Processor 280 Step 284 Step 286 Step 288 Step 290 Step 292 Step 294 Step 296 Step 298 Step 300 Step 302 Step 304 Disposable Structure 306 Annular Concentrate Dam 308 Tapered Outer Surface 310 Separate Liquid Dam 312 Annular Recess 314 Disposable Centrifuge Structure 316 Separation liquid pump chamber 318 Lower separation liquid centripetal pump chamber surface 320 Upper separation liquid centripetal pump chamber surface 322 Lower separation liquid centripetal pump chamber opening 324 Upper separation liquid centripetal pump chamber opening 326 Lower separation liquid chamber blade 328 Upper separation liquid chamber blade 330 Separation Liquid centripetal pump opening 332 Concentrate pump chamber 334 Lower concentrate centripetal pump chamber surface 336 Upper separated liquid centripetal pump chamber surface 338 Lower concentrate centripetal pump chamber opening 340 Lower concentrate chamber vane 342 Upper concentrate centripetal pump chamber opening 344 Upper concentrate Chamber vane 346 Concentrate pump opening 348 Air/liquid interface 350 Air/liquid interface 352 Disposable structure 354 Air line 356 Opening 358 System 360 Back pressure regulator 362 Pressurized air source 364 Pressure control pilot valve 366 Pilot line 368 Pressure control valve 370 Disposable structure 372 Upper disk-like portion 374 Isolate centripetal chamber 376 Concentrate centripetal chamber 377 Annular edge 378 Circular partition wall 379 Surface 380 Cell concentrate channel 381 Funnel channel 382 Cell concentrate channel inlet 383 Cell concentrate guide surface 384 Cell concentrate 386 Cell concentrate outlet 388 Concentrate centripetal pump inlet 390 Tapered portion 392 Radially extending portion 394 Upper piece 396 Lower piece 398 Upper annular interface 400 Lower annular interface 402 Conical annular upper surface 404 Top portion 406 Conical annular bottom portion 408 Bottom portion 410 Separator channel 412 Inlet 414 Disposable structure 416 Top disc-shaped portion 418 Outer wall 420 Bottom portion 422 Cylindrical core 424 Outer partition wall 426 Vanes 428 Axis 430 Supply tubing 432 Tubing opening 434 Cylindrical shaped opening 436 separated liquid outlet line 438 separated liquid centripetal pump 440 separated liquid centripetal pump chamber 442 separated liquid centripetal pump chamber 442 separated liquid chamber 444 separated liquid channel inlet 446 separated liquid channel 448 condensate outlet line 450 concentrated liquid centripetal pump 452 concentrated liquid centripetal pump chamber 454 concentrated liquid channel 456 top piece 458 bottom piece 460 bottom piece 462 concentrate channel inlet 464 vertical opening 468 tapered portion 470 portion 1000 centrifuge structure 1100 flexible liner 1110 thermal bond 1200 bottom flange 1210 top surface 1300 top flange 1310 bottom surface 1400 centripetal pump 1410 Pairing Disc 1415 Gap 1420 Round Pump Room 1420 Rotation Pump Room 1500 Core Structure 1505 Top 1515 Outer 1520 Cavity 1525 Rotation Axis 1530 Slits / Pole 15550 Separation Room 1555 Separation Rules 1560 Pretend 1580 Plate 1580 Plate 1580 Plate 1590 accelerator bowl 1600 centrifugal separator cover 1605, 1620 inner surface 1610 engagement cap portion 1630 radial fin 1640 wall portion 1700 rotary mechanical seal 2000 feed/discharge device 2100 feed pipe 2110 nozzle 2200 separated liquid discharge 2300 feed pipe carrier 2400 separated liquid outlet 2500 concentrated liquid Exhaust 2500 Cell Ejection Tube 2510 Peristaltic Pump 3000 Multi-Use Structure 3100 Bowl 3200 Multi-Use Bowl Cover 3210 Nabin 4400 Centripetal Pump 4410 Pairing Disc 4415 Gap 4420 Pump Chamber 4430 Concentration Sensor 4540 Slit/hole 4630 Accelerator Fins 5000 Diluent Supply 5100 Disk channel 5150 Diluent pump 6000 Distance 6010 Depth 6100 Throttle tube R Arrow V Distance

Claims (24)

A structure for releasably receiving within a rotatable centrifuge bowl, the structure being provided within the bowl for operating cells in a cell culture into a cell concentrate and a cell dissociate within an interior region of said structure. A device having a structure that effectively isolates
In the operating position the structure comprises an upper disc-shaped portion;
a lower part;
a cylindrical core vertically intermediate said upper disc-shaped portion and said lower portion;
a separation chamber radially outward from and in surrounding relation to said core;
an outer wall configured to operatively engage the bowl, comprising:
extending in liquid-tight relation to said upper disc-shaped portion and bounded by said separation chamber;
extending in a relationship surrounding the core and the separation chamber;
and an outer wall having an inner shape of a truncated cone, the inner radius adjacent to the lower portion being smaller than the inner radius adjacent to the upper disc-shaped portion;
a vertically extending supply pipe;
a vertically extending separated liquid discharge pipe;
a vertically extending concentrate discharge line;
has
rotatable about a vertical axis with said upper disc-shaped portion and said outer wall in operative engagement with said bowl;
said structure further comprising a separation liquid centripetal pump axially aligned with said core and coaxially positioned about said supply line and in fluid communication with said separation liquid discharge line;
said separation liquid centripetal is located in a separation liquid centripetal chamber within said upper disc-shaped portion;
said separation fluid centripetal chamber is in fluid communication with said separation chamber via at least one separation fluid channel extending into said upper disc-shaped portion;
each separation liquid channel extending in fluid communication between a separation liquid channel inlet located respectively radially outwardly of the core and the separation liquid centripetal chamber;
The structure is further axially aligned with the core, coaxially positioned about the supply line, vertically above the separate liquid centripetal pump, and in fluid communication with the concentrate discharge line. having a concentrate centripetal pump that
said concentrate centripetal pump is located in a concentrate centripetal chamber within said upper disc-shaped portion;
said concentrate centripetal pumping chamber is in fluid communication with said separation chamber via at least one radially extending concentrate channel extending within said upper disc-shaped portion;
said at least one radially extending concentrate channel extending radially between each radially outwardly positioned concentrate channel inlet of every single separate liquid inlet;
when the bowl rotates, the upper disc-shaped portion and the outer wall rotate relative to each of the feed line, the separated liquid discharge line, the concentrate discharge line, the separated liquid centripetal pump and the concentrate centripetal pump;
each concentrate channel having a respective concentrate channel inlet and a channel portion intermediate said concentrate centripetal chamber, the direction of concentrate flow within the respective concentrate channel portion area of said concentrate centripetal chamber; A device characterized by a continuously increasing cross-sectional area perpendicular to the axis, with a corresponding increase in radial proximity to said axis.
2. The apparatus of claim 1, wherein said channel portion of each said concentrate channel begins at said respective concentrate channel inlet and extends radially inward.
2. The apparatus of claim 1, wherein said channel portion of each said concentrate channel begins at said respective concentrate channel inlet and extends radially inwardly and upwardly from said concentrate channel inlet.
said channel portion of each said concentrate channel beginning at said respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
2. Each of said concentrate channels further comprises a horizontally radially extending portion, said horizontally radially extending portion extending fluidly intermediate said channel portion and said concentrate centripetal chamber. The apparatus described in .
said channel portion of each said concentrate channel beginning at said respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
each of said concentrate channels further having a horizontally radially extending portion extending fluidly intermediate said channel portion and said concentrate centripetal chamber;
a horizontally radially extending portion of each of said concentrate channels terminating radially inward at a respective cell concentrate channel outlet within said concentrate centripetal pump chamber;
2. The apparatus of claim 1, wherein the horizontally radially extending portion of each concentrate channel has a constant cross-sectional area along its length.
said channel portion of each said concentrate channel beginning at said respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
2. The apparatus of claim 1, wherein each of said concentrate channels further has a horizontally radially extending portion, said horizontally radially extending portion extending radially outwardly from said concentrate centripetal chamber. .
said channel portion of each said concentrate channel beginning at said respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
each of said concentrate channels further having a horizontally radially extending portion, said horizontally radially extending portion extending outwardly from said concentrate centripetal chamber;
said channel portion terminating in said horizontally and radially extending portion;
2. The apparatus of claim 1, wherein the horizontally and radially extending portion of each concentrate channel has a constant cross-sectional area along its length.
said channel portion of each said concentrate channel beginning at said respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
Each of said concentrate channels further has a horizontally radially extending portion extending outwardly from a respective cell concentrate channel outlet into said concentrate centripetal chamber. ,
said concentrate centripetal pump having a concentrate centripetal pump inlet, each cell concentrate channel outlet being axially and radially aligned with said concentrate centripetal pump inlet;
each channel portion terminating radially inwardly at a respective horizontally radially extending portion;
2. The apparatus of claim 1, wherein said horizontally and radially extending portion of each said concentrate channel has a constant cross-sectional area along its length.
said upper disc-shaped portion having an upper piece and a lower piece in mating relationship therewith;
2. The apparatus of claim 1, wherein each concentrate channel is bounded by at least one lower surface of the upper piece and at least one upper surface of the lower piece.
2. The apparatus of claim 1, wherein said at least one radially extending concentrate channel comprises a unitary, substantially annular concentrate channel.
2. The apparatus of claim 1, wherein said at least one radially extending concentrate channel comprises a plurality of angularly spaced individual concentrate channels.
said at least one radially extending concentrate channel comprises a single, substantially annular concentrate channel;
2. The apparatus of claim 1, wherein said channel portion of said concentrate channel begins at a substantially annular concentrate channel inlet and extends radially inwardly and upwardly from said concentrate channel inlet.
said at least one radially extending concentrate channel comprises a single, substantially annular concentrate channel;
said channel portion of said concentrate channel beginning at a substantially annular concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
The concentrate channel further has a substantially annular, horizontally radially extending portion that extends outwardly from the substantially annular cell concentrate channel outlet to the concentrate. extending to the liquid centripetal pump chamber,
2. The device of claim 1, wherein the channel portion terminates radially inwardly at said horizontal, radially extending portion.
said at least one radially extending concentrate channel comprises a single, substantially annular concentrate channel;
a channel portion of said concentrate channel beginning at a substantially annular concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
2. The apparatus of claim 1, wherein said upper disc-shaped portion has a substantially annular funnel channel, said annular funnel channel extending upwardly and radially inward to said annular concentrate channel inlet.
said at least one radially extending concentrate channel comprises a single, substantially annular concentrate channel;
said channel portion of said concentrate channel beginning at a substantially annular concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
said upper disc-shaped portion having a substantially annular funnel channel, said annular funnel channel extending upwardly and radially inward to said annular concentrate channel inlet;
The upper disc-shaped portion has a substantially annular cell concentrate guiding surface that extends below the annular funnel channel and bounds radially outwardly with the separation chamber. in contact with
2. The apparatus of claim 1, wherein said annular cell concentrate guiding surface further extends radially outwardly and upwardly adjacent said annular funnel channel.
said at least one radially extending concentrate channel comprises a single, substantially annular concentrate channel;
a channel portion of said concentrate channel beginning at a substantially annular concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
said upper disc-shaped portion having a substantially annular funnel channel extending upwardly and radially inward to said annular concentrate channel inlet;
The upper disc-shaped portion has a substantially annular cell concentrate guiding surface that extends below the annular funnel channel and bounds radially outwardly with the separation chamber. in contact with
said annular cell concentrate guiding surface extending further radially outward and upwardly adjacent said annular funnel channel;
said upper disc-shaped portion bounded by a radially extending surface on the underside of said separation chamber, said radially extending surface terminating radially outwardly at a substantially annular edge, said 2. The apparatus of claim 1, wherein an annular edge is axially above said radially outwardly extending annular cell concentrate guiding surface and said annular funnel channel extends upwardly from said annular edge. .
said at least one radially extending concentrate channel having a plurality of angularly spaced individual concentrate channels;
a channel portion of each of said concentrate channels beginning at a respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
2. Apparatus according to claim 1, having a constant cross-sectional width in a direction perpendicular to the direction of concentrate flow and a vertical height that varies according to radial distance from said axis.
said at least one radially extending concentrate channel having a plurality of angularly spaced individual concentrate channels;
said channel portion of each said concentrate channel beginning at a respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
having a constant cross-sectional width in a direction perpendicular to the direction of concentrate flow and a vertical height that varies according to radial distance from said axis;
2. The apparatus of claim 1, wherein said upper disc-shaped portion has a plurality of angularly spaced vertical openings, each vertical opening extending between the radial perimeter of said separation chamber and a respective channel entrance.
said at least one radially extending concentrate channel having a plurality of angularly spaced individual concentrate channels;
said channel portion of each said concentrate channel beginning at a respective concentrate channel inlet and extending radially inwardly and upwardly from said concentrate channel inlet;
having a constant cross-sectional width in a direction perpendicular to the direction of concentrate flow and a vertical height that varies according to radial distance from said axis;
2. The device of claim 1, wherein the upper disc-shaped portion has an upper piece and a lower piece in engagement, each concentrate channel bounding a surface of each of the upper and lower pieces.
2. The device of claim 1, wherein said structure is a disposable structure.
A structure for releasably receiving within a rotatable centrifuge bowl, the structure being provided within the bowl for operating cells in a cell culture into a cell concentrate and a cell dissociate within an interior region of said structure. A device having a structure that effectively isolates
The structure, in the operating position, has an upper disc-shaped portion and
a cylindrical core extending vertically under the upper disc-shaped portion;
an outer wall configured to operatively engage the bowl, comprising:
extending in liquid-tight engagement with said upper disc-shaped portion;
extending in surrounding relation to said core;
the shape at the end of the structure vertically spaced from the upper disc-like portion is frusto-conical with a shorter inner radius;
an outer wall extending in surrounding relation to said core and bounding said separation chamber within said structure extending radially intermediate said core and said outer wall;
a vertically extending cell culture feed line;
a vertically extending separated liquid discharge pipe;
a vertically extending concentrate discharge line;
has
said upper disc-shaped portion and said outer wall being operatively engaged with said bowl and rotatable about a vertical axis; coaxial about a vertical axis,
Said upper disc-shaped portion comprises:
a separation liquid centripetal pump chamber in fluid communication with said separation chamber via at least one separation liquid opening;
a concentrate centripetal pump chamber in fluid communication with said separation chamber via at least one concentrate channel, said at least one concentrate channel comprising:
having a concentrate channel inlet located radially outwardly from the at least one separate liquid opening;
extending radially and fluidly between the at least one concentrate inlet and the concentrate centripetal chamber;
the structure further includes a separation liquid centripetal pump coaxially positioned with respect to the supply line within the separation liquid centripetal chamber and in fluid communication with the separation liquid discharge line;
A concentrate centripetal pump coaxially positioned within the concentrate centripetal chamber with respect to the supply line, vertically above the separate liquid centripetal pump and in fluid communication with the concentrate outlet line. a concentrate centripetal pump;
when the bowl rotates, the upper disc-shaped portion and the outer wall rotate relative to each of the feed line, the separated liquid discharge line, the concentrate discharge line, the separated liquid centripetal pump and the concentrate centripetal pump;
Each concentrate channel has a channel portion extending intermediate said respective concentrate channel inlet and said concentrate centripetal chamber, said channel portion being oriented in the direction of concentrate flow within said respective concentrate channel portion. A device characterized by a continuously increasing cross-sectional area perpendicular to the axis with a corresponding increase in radial proximity to said axis.
22. The apparatus of claim 21, wherein each concentrate channel portion begins at a respective concentrate channel inlet of a respective concentrate channel and extends radially inwardly and upwardly from the respective channel inlet.
23. The apparatus of claim 22, wherein said at least one channel comprises a single, substantially annular concentrate channel.
23. The apparatus of claim 22, wherein said at least one channel comprises a plurality of angularly spaced individual concentrate channels.

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