Apparatus, system, and method for filtering waste in rocker bioreactors

Abstract

The bioprocessing device includes a vessel having a flexible exterior defining an internal cavity configured to receive a bioprocessing fluid, the vessel configured for selective attachment to a bioreactor. The vessel includes at least one filter within the internal cavity to retain cells within the vessel while waste is extracted from the internal cavity, and the flexible exterior defines a plurality of waste ports, each of which is fluidly connected to at least one filter to allow waste to be extracted from the vessel. The vessel is configured to reduce the likelihood of the filter clogging when high cell densities occur during bioprocessing within the vessel.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION Embodiments of the present invention relate generally to bioprocessing devices, systems, and methods, and more particularly to filtering waste from a rocker bioreactor vessel during a perfusion process. [Background technology]

[0002] Bioreactors are often used to carry out biochemical or biological processes and to manipulate the liquids and other products of such processes. Such bioreactors often contain flexible or collapsible vessels, such as disposable bags supported by an outer rigid structure. These "disposable" bioreactors include stirred-tank reactors, in which a sterile disposable bag and agitator are housed within a rigid tank, and rocker bioreactors, in which the disposable bag is secured to a rocking platform.

[0003] In a rocker bioreactor, disposable bags are secured to a tray, which is connected to a motorized rocking device. The rocking device rotates the tray back and forth around an axis / pivot point. The motion caused by the rocking device creates waves in the cell culture medium inside the bag. The waves mix the oxygen supplied to the bag, transferring it from a gas to a liquid, creating an ideal environment for cell growth. To monitor and promote cell growth, rocker bioreactors contain pumps that, among other functions, add fresh cell growth / expansion medium to the bag and remove spent medium from the bag.

[0004] In certain known rocker bioreactors, removal of spent medium is accomplished by a pump that draws the medium from the bag and sends it to a waste container through external tubing connected to a waste port in the bag wall. In perfusion processes, which provide a continuous supply of fresh nutrients while simultaneously removing spent medium and waste products, filters are used to retain the cells in the bag while waste products are removed.

[0005] Referring to FIG. 3, such filters, e.g., filter 38, typically rest or float on top of the cell culture medium 32 within bag 30 and have their own tubing 40 that connects to a waste port inside the bag. These filters contain a series of small openings or pores on the side facing the cell culture medium, sized to prevent cells from passing through the filter and out of the bag while allowing spent medium and other waste products to be removed. The opposite side of the filter typically includes a solid, non-porous surface onto which internal tubing 40 is formed or attached. In use, a pump applies suction to filter 38 through an external line connected to the waste port, drawing waste from bag 30 while retaining cells within the filter's pores.

[0006] However, in known bags, the filter can become clogged at high cell densities (e.g., above about 1e7 cells / mL). In particular, such filters can become clogged in and around the area where the inner tube 40 is formed or attached to the filter 38. This clogging can occur even when other parts of the filter are free of clogging, resulting in a filter that is easily clogged and functionally much smaller than its overall footprint.

[0007] Naturally, such clogging can have a detrimental effect on cell growth, as it prevents spent media and other waste products from being removed from the bag. This is generally undesirable, as biologics are time-consuming to manufacture and highly valuable. Indeed, this can be particularly serious in the case of cell therapies, where the cellular contents of the bag may be needed urgently.

[0008] In view of the above, there is a need for a rocker bioreactor vessel that can reduce or prevent the likelihood of waste filter clogging at high cell densities, particularly during continuous and / or perfusion bioprocessing. Summary of the Invention

[0009] Certain embodiments commensurate in scope with the originally claimed subject matter are summarized below. These embodiments are not intended to limit the scope of the claimed subject matter; rather, these embodiments are intended merely to provide a brief summary of possible embodiments. Indeed, this disclosure may encompass a variety of forms that may be similar to or different from the embodiments set forth below.

[0010] According to one aspect of the present invention, a bioprocessing apparatus includes a vessel having a flexible exterior defining an internal cavity configured to receive a bioprocessing fluid, the vessel configured for selective attachment to a bioreactor. The apparatus further includes at least one filter within the internal cavity for retaining cells within the vessel while waste material is extracted from the internal cavity, and a plurality of waste ports formed on the flexible exterior, each of the plurality of waste ports fluidly connected to the at least one filter to allow waste material to be extracted from the vessel. The vessel is configured to reduce the likelihood of the filter becoming clogged when high cell densities occur during bioprocessing within the vessel.

[0011] In one embodiment, the bioreactor is a rocker bioreactor and the bioprocessing is a continuous perfusion cell culture process.

[0012] In one embodiment, the device further comprises a stopcock manifold fluidly connected to the plurality of waste ports via external waste lines, the stopcock manifold allowing a user to select an unclogged portion of the at least one filter in the event that another portion of the at least one filter becomes clogged.

[0013] In one embodiment, the at least one filter is a plurality of filters, and each of the plurality of waste ports is fluidly connected to a separate filter of the plurality of filters so as to allow waste to be extracted from the container.

[0014] In one embodiment, the multiple filters are separate filter membranes formed on a single piece of material.

[0015] In one embodiment, the device further comprises a manifold fluidly connected to the plurality of waste ports via external waste lines, the manifold allowing for simultaneous extraction of waste through each of the plurality of filters.

[0016] In one embodiment, the manifold may be a stopcock manifold that allows a user to select an unclogged filter from the plurality of filters in the event that another filter from the plurality of filters becomes clogged.

[0017] In one embodiment, each of the plurality of filters is fluidly connected to a waste port via a filter line disposed within the interior cavity.

[0018] In one embodiment, the multiple filters can be two filters each fluidly connected to a separate waste port via a filter line.

[0019] In one embodiment, the plurality of filters is four filters, each filter fluidly connected to a separate waste port via a filter line.

[0020] In one embodiment, the at least one filter may include pores having a pore size of about 1.2 μm.

[0021] In one embodiment, the high cell density may be greater than about 1e7 cells / mL.

[0022] In one embodiment, multiple filters are secured within the interior cavity.

[0023] According to another aspect of the present invention, a bioprocessing system includes a vessel having a flexible exterior defining an internal cavity configured to receive a bioprocessing fluid, the vessel configured to be selectively attached to a rocker bioreactor, and a plurality of filters within the internal cavity for retaining cells within the vessel during a continuous perfusion cell culture process. The system further includes a plurality of waste ports formed on the flexible exterior, the plurality of waste ports fluidly connected to the plurality of filters via filter lines disposed within the internal cavity to allow waste to be extracted from the vessel, and a manifold fluidly connected to the plurality of waste ports via external waste lines. The system also includes a waste bag fluidly connected to the manifold for receiving waste from the vessel during the continuous perfusion cell culture process. The manifold allows waste to be extracted simultaneously through each of the plurality of filters.

[0024] In one embodiment, the manifold is a stopcock manifold that allows a user to select an unclogged filter from the plurality of filters if another filter from the plurality of filters becomes clogged during a continuous perfusion cell culture process.

[0025] In one embodiment, the multiple filters may be multiple separate filter membranes formed on a single piece of material.

[0026] In one embodiment, the multiple filters can be two filters each fluidly connected to a separate waste port via a filter line.

[0027] In one embodiment, the plurality of filters can be four filters, each fluidly connected to a separate waste port via a filter line.

[0028] In one embodiment, each of the plurality of filters may include pores having a pore size of about 1.2 μm.

[0029] In one embodiment, the high cell density may be greater than about 1e7 cells / mL.

[0030] According to another aspect of the present invention, a bioprocessing vessel includes a vessel having a flexible exterior defining an internal cavity configured to receive a bioprocessing fluid, the vessel configured for selective attachment to a bioreactor. The vessel further includes a plurality of filters within the internal cavity for retaining cells within the vessel while waste is extracted from the internal cavity, and a manifold disposed on the vessel, the manifold having a plurality of filter line ports within the internal cavity and at least one waste port on the flexible exterior. The vessel further includes a plurality of filter lines fluidly connecting the plurality of filters to the plurality of filter line ports on the manifold, thereby allowing waste to be extracted from the vessel via the at least one waste port. The vessel is configured to reduce the likelihood of the filters clogging when cell densities are high during bioprocessing within the vessel.

[0031] In one embodiment, the manifold may be a stopcock manifold that allows a user to select an unclogged filter from the plurality of filters in the event that another filter from the plurality of filters becomes clogged at high cell densities during bioprocessing. [Brief explanation of the drawings]

[0032] The invention will be better understood from reading the following description of non-limiting embodiments, with reference to the accompanying drawings, in which:

[0033] [Figure 1] FIG. 1 is a perspective view of a rocker bioreactor system suitable for use in embodiments of the present invention. [Figure 2] 2 is a perspective view of a known bioreactor vessel for use in conjunction with the rocker bioreactor of FIG. 1, showing the filter of the vessel. [Figure 3] FIG. 3 is a side view of the bioreactor vessel / bag of FIG. 2. [Figure 4]FIG. 1 is a perspective view of a bioreactor vessel and filter according to one embodiment of the present invention. [Figure 5] FIG. 1 is a perspective view of a bioreactor vessel and filter according to an alternative embodiment of the present invention. [Figure 6] FIG. 1 is a perspective view of a bioreactor vessel and filter according to another embodiment of the present invention. [Figure 7] FIG. 1 is a schematic diagram of a bioreactor vessel, filter, and associated equipment according to one embodiment of the present invention. [Figure 8] FIG. 1 is a schematic diagram of a bioreactor vessel, filters, and associated equipment showing four filter and port configurations according to one embodiment of the present invention. [Figure 9] 9 is a schematic diagram of the filter and port configuration of FIG. 8, where the filter is mounted on a plate fixed to the bottom surface of the vessel in accordance with an embodiment of the present invention. [Figure 10] 9 is a schematic diagram of the filter and port configuration of FIG. 8 with the filter secured to the bottom surface of the vessel, according to another embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0034] Reference will now be made in detail to the exemplary embodiments of the invention, examples of which are illustrated in the accompanying drawings. Wherever possible, the same reference numbers will be used throughout the drawings to refer to the same or like parts.

[0035] As used herein, the terms "flexible" or "foldable" refer to a structure or material that is flexible or that can be bent without breaking, and may also refer to compressible or expandable materials. An example of a flexible structure is a bag made of polyethylene film. As used herein, the terms "rigid" and / or "semi-rigid" are used interchangeably to describe a "non-collapsible" structure, i.e., a structure that does not fold, bend, or otherwise deform under normal force and that has a significantly reduced elongated dimension.

[0036] As used herein, the term "vessel" refers to a flexible bag, flexible vessel, semi-rigid vessel, or rigid vessel, as the case may be. The term "vessel" as used herein is intended to encompass bioreactor vessels, where the walls or portions of the walls are flexible or semi-rigid, disposable flexible bags, and other vessels or conduits commonly used in biological or biochemical processes, including, for example, cell culture / purification systems, fermentation systems, mixing systems, media / buffer preparation systems, and filtering / purification systems.

[0037] As used herein, the term "bag" refers to a flexible or semi-rigid container or vessel used, for example, as a bioreactor or mixer to house contents. Although embodiments of the present invention are described for use with bioprocessing bags, including but not limited to bioreactor bags and mixer bags, embodiments may be configured for use with other bags or vessels.

[0038] Embodiments may be used to perform a variety of bioprocesses within rocker bioreactors, including but not limited to cell growth. Certain embodiments may find broad application in bioprocessing and biochemical environments, and may potentially be used in non-biological / biochemical contexts as well. Similarly, while embodiments are described and illustrated with respect to specific rocker bioreactor systems and in connection with continuous perfusion cell culture processes, embodiments may also be utilized in other types of reactor / mixers, particularly those equipped with vessels in which continuous cell culture occurs and / or where waste is removed via filters.

[0039] The embodiments are similarly not limited to any particular size / shape of container / bag, and although the embodiments are particularly suitable for and / or described for use with 20 L to 50 L flexible bags, other sizes, e.g., 2 L to 10 L, may also be used.

[0040] 1, an exemplary rocker bioreactor system 10 suitable for use in embodiments of the present invention is shown. As shown, the system 10 includes a bioreactor 12 connected to one or more peristaltic pumps 14. The bioreactor 12 is also operably connected to a controller, such as a local or remote computer, that provides process / protocol monitoring, etc., via a wired or wireless connection (not shown).

[0041] Bioreactor 12 includes a removable tray 20 configured to selectively receive and support vessels / bags. In an embodiment, tray 20 includes a removable lid 28 with a hinged door 30 that can be lifted to access tray 20 and the vessels secured thereto. Bioreactor 12 also includes a base 16 operatively connected to tray 20. Base 16 houses a rotation mechanism, e.g., a motor, that can rotate tray 20 back and forth.

[0042] In use, the containers / bags are secured to the tray 20 via one or more attachment mechanisms, such as selectively lockable clips at either end of the tray 20. The tray 20 and bag are then rotated back and forth, creating a rocking motion that induces waves in the cell culture medium within the bag to promote growth.

[0043] 2 and 3, a conventional flexible reactor vessel / bag 30 is shown containing a fluid 32 containing growth medium therein. Removal of spent medium is accomplished by a pump 14, such as a peristaltic pump, which draws the medium from the bag 30 and discharges it into a waste container via an external line / tubing 36 connected to a waste port 34 in the bag wall. To provide perfusion, for example, a filter 38 is positioned within the bag 30 to retain the cells within the bag 30 while removing the spent fluid medium 32. The filter 38 has its own filter line / tubing 40 inside the bag, which is also connected to the waste port 34. However, known filters can only remove a minimum of about 1e7 cells / mL (1 x 10 7 ) may cause clogging.

[0044] In particular, filter 38 may become clogged in and around the area where inner tube 40 is formed or attached to filter 38. This clog may exist even when other portions of filter 38 are unclogged, resulting in filter 38 easily becoming clogged and functionally smaller than its overall footprint. As will be appreciated, such clogs can have a detrimental effect on cell growth by preventing spent media and other waste products from being removed from bag 30.

[0045] Referring to Figure 4, a bioprocessing apparatus according to one embodiment of the present invention is shown. As shown, the apparatus includes a bag / container 130 having a flexible exterior defining an interior cavity configured to contain a bioprocessing fluid 132, such as cell culture media. The container 130 is configured to be selectively attached to a tray 20 of a rocker bioreactor 16.

[0046] As shown, the interior cavity of the container 130 includes at least one filter 138 and a plurality of waste ports 134, 137. In the illustrated embodiment, the filter 138 extends transversely relative to the bag 30 and is connected to a plurality of waste ports 134, 137, which are fluidly connected to a waste bag (not shown) via a plurality of internal tubing / filter lines 140, 141 and external waste lines 136, 139. As will be appreciated, the filter lines 140, 141 are fluidly connected to the filter 138 such that waste that passes through the pores on the cell culture medium-facing side of the filter (not shown) exits the container 130 via the waste ports 134, 137.

[0047] The waste ports 134, 137 are formed on the flexible exterior surface of the container 130. As will be appreciated, the waste ports 134, 137 may be welded to the container 130, molded into the container 130, or otherwise integrated with the container 130. However, embodiments are not limited to a particular size, shape, or configuration of the waste ports 134, 137; conventional ports may be suitable, and commercially available external waste lines 136, 139 and waste bags may be used. The location of the waste ports 134, 137 may vary, but in embodiments, they may be located near (e.g., directly above) where the filter lines 140, 141 connect to the at least one filter 138.

[0048] The filter 138 has numerous pores (not shown) on the side facing the cell culture medium. The opposite side of the filter 138 is solid and non-porous, and has filter lines 140, 141 attached or formed thereto. In embodiments, the filter 138 is a laminate or composite of porous and non-porous materials with a mesh spacer disposed between them to prevent the porous and non-porous materials from collapsing together when suction is applied. The filter lines 140, 141 can be secured to the filter in a variety of ways, such as by heat welding or molding.

[0049] It will be appreciated that embodiments are not limited to a particular filter material, structure, or pore size, although in particular embodiments, the filter may be fabricated from a polymer such as polyvinylidene fluoride (PVDF) or polysulfone (PES) and have a pore size of approximately 1.2 μm. Similarly, the filter lines 140, 141 are not limited to a particular polymer material, inner or outer diameter, length, or shape.

[0050] In certain embodiments, filter 138 may be connected to two or more filter lines. For example, filter 138 may be connected to a filter line located at each corner of filter 138, or in an array or other arrangement along filter 138. Indeed, while embodiments utilize at least two filter lines, the number of filter lines is not limited to only two, and is determined by various factors, such as the amount of space available for corresponding waste ports on the flexible exterior of the bag. In certain embodiments, four filter lines (and four exhaust ports) may be used (see Figures 8-10).

[0051] Similarly, embodiments are not limited to any particular shape or size of filter, although rectangular filters are common. It will be appreciated that other filter shapes may be utilized without departing from the invention. In embodiments, filter 138 is approximately 7 inches by 7 inches (17.8 cm by 17.8 cm) for 2 L and 10 L containers / bags and approximately 15 inches by 15 inches (38.1 cm by 38.1 cm) for 20 L and 50 L containers / bags, although other sizes may be used.

[0052] The container / bag itself can be manufactured from a variety of materials, and the invention is not limited in this regard. In embodiments, the container can be a multi-layer laminate of USP Class VI material. The container can be fully or partially transparent to allow observation of the internal cavity.

[0053] 4, filter lines 140, 141 reduce the likelihood of clogging by providing suction at multiple locations on filter 138. In this manner, a greater portion of filter 138 may be effectively utilized. Additionally, as described in more detail below, in certain embodiments, one or more of filter lines 140, 141 may be selectively activated / deactivated, for example, via a stopcock manifold, allowing a user to select an unclogged portion of the filter for use when another portion becomes clogged.

[0054] Referring now to Figure 5, an embodiment is also available that includes a longitudinally extending filter 238. This embodiment is similar to the embodiment of Figure 4, except for the orientation of the filter 238 and the location of the waste port 247. That is, the device includes a container 230 with a filter 238 within its internal cavity. The filter 238 is fluidly connected to waste ports 234 and 237 via filter lines 240 and 241, respectively. Waste lines 236 and 239 extend from the waste ports 234 and 237 to waste bags (not shown).

[0055] Referring to Figure 6, another embodiment of the present invention is shown. In this embodiment, a container 330 has a filter that includes multiple filters, which are separate filter membranes 335, 338 formed on a single piece or sheet of material (e.g., a polymer laminate). As used herein, the term "filter" includes such filter membranes as well as physically separated filters. Each filter membrane 335, 338 has pores on the side facing the cell culture medium (not shown). The filter membranes 335, 338 are separated by a section 350 that effectively separates or seals the filter membranes 335, 338. This can be achieved in a variety of ways; in one embodiment, the section 350 is a heat seal between the filter membranes 335, 338.

[0056] Each of the filter membranes 335, 338 is connected to its own internal (e.g., first and second) filter line 340, 341 and waste port 334, 337. The first filter line 340 and second filter line 341 use separate (e.g., first and second) waste lines 336, 339 operably connected to a pump and waste bag.

[0057] In this embodiment (and the embodiment shown in FIG. 7), it is envisioned that the filter membranes 335, 338 may be used independently of one another via a stopcock manifold or other switching device, allowing a user to select an unclogged filter from the plurality of filters in the event that another filter from the plurality becomes clogged.

[0058] As will be appreciated, in certain embodiments, multiple filters may be attached to each other (or to specific filters of the multiple filters) via other structures or mechanisms, i.e., the filter membranes need not be formed from a single sheet of material, but may be completely separate filters welded together at specific locations.

[0059] 7, a system 300 for bioprocessing includes a vessel 360 having a flexible exterior defining an interior cavity. The interior cavity contains a plurality of filters 362, 364. Here, the plurality of filters 362, 364 are two filters that are physically separated and are not filter membranes on a single piece of sheet material.

[0060] Filters 362, 364 are fluidly connected to multiple waste ports 370, 372 via internal filter lines 366, 368. Each filter 362, 364 is connected to a separate waste port via a filter line 366, 368. External waste lines 374, 376 are connected to waste ports 370, 372 and to a three-way valve / stopcock manifold 378, which is further connected to a peristaltic pump 382 and waste bag 348 via lines / tubing 380, 384. As previously mentioned, in this embodiment, clogged filters can be replaced with unused filters during the bioprocessing process via stopcock manifold 342.

[0061] Referring now to Figure 8, in certain embodiments, a system 400 for bioprocessing includes a vessel 430 with a flexible exterior defining an interior cavity containing multiple filters (e.g., filter membranes 420, 422, 424, 426), the filters being four filters formed on a single piece of material (e.g., a polymer laminate). As with the embodiment of Figure 6, the filter membranes 420, 422, 424, 426 are separated from one another by heat seals or other structures, represented schematically by the illustrated grid. As with the other embodiments, the filter membranes 420, 422, 424, 426 have pores that include a fluid-facing side that contacts the cell culture medium when the filter is suspended on the cell culture medium.

[0062] In this embodiment, each filter membrane 420, 422, 424, 426 includes a filter line internal to the vessel, not shown but configured according to the previously described embodiment. One filter line fluidly connects each filter membrane 420, 422, 424, 426 to a plurality of waste ports (e.g., four waste ports 431, 433, 435, 437). Each filter membrane 420, 422, 424, 426 is connected to a separate waste port 431, 433, 435, 437 via a filter line. External waste lines 432, 434, 436, 439 are connected to the waste ports 431, 433, 435, 437 and are connected to a four-way manifold 442, which is connected via lines / tubing to a peristaltic pump 446 and a waste bag 448. As will be appreciated, in this embodiment, waste may be extracted from all four filter membranes simultaneously via manifold 442 .

[0063] In an embodiment, manifold 442 is a stopcock manifold (or other switching manifold or device) that allows for replacement of clogged filter membranes with unused or known unclogged filter membranes midway through a bioprocessing run.

[0064] Referring to Figure 9, in this embodiment, a system 500 for bioprocessing includes a vessel 530 having a flexible exterior defining an interior cavity, the interior cavity containing a plurality of filters, e.g., four filter membranes 520, 522, 524, 526 formed on a single piece of material, such as a polymer laminate. Similar to the embodiment of Figure 8, the filter membranes 520, 522, 524, 526 are separated from one another by heat seals or other structures, represented schematically by the illustrated grid. Additionally, the filter membranes 520, 522, 524, 526 have pores that include fluid-facing sides that contact the cell culture medium when the filters are suspended on the medium.

[0065] Each filter membrane 520, 522, 524, 526 includes a filter line that fluidly connects the membrane to multiple waste ports (e.g., four waste ports 531, 533, 535, 537), external waste lines 532, 534, 536, 539, a four-way manifold 542 (which in embodiments may be a stopcock manifold), a pump 546, and a waste bag 548 via lines / tubing. However, in this embodiment, the filter membranes 520, 522, 524, 526 are secured to a rigid plate 552 that is secured to the bottom of the vessel.

[0066] Similarly, as shown in Figure 10, a bioprocessing system 600 includes a vessel 630 containing four filter membranes 620, 622, 624, and 626 formed on a single piece of material, and includes a filter line fluidly connecting the membranes to multiple waste ports (e.g., four waste ports 631, 633, 635, and 637), external waste lines 632, 634, 636, and 639, a four-way manifold 642 (e.g., a stopcock manifold), a pump 646, and a waste bag 648 via lines / tubing. However, in this embodiment, the filter membranes 620, 622, 624, and 626 are not attached to a rigid plate as in the embodiment of Figure 9, but are instead secured to the bottom of the vessel via multiple tethers. The tethers may be formed from the same single piece of material as the filter membranes or may be welded to the bottom of the vessel. Of course, other attachment mechanisms may be employed without departing from the scope of the present invention.

[0067] Although embodiments are described for use in connection with manifolds and stopcock manifolds, in general, a variety of manifolds or switching mechanisms / valve banks may be utilized, including pneumatic valves, etc. In embodiments, such manifolds / switching mechanisms may be operably connected to a controller.

[0068] In certain embodiments, the container may include a manifold on the container itself. In such embodiments, the manifold is disposed on the flexible exterior surface of the container (e.g., formed within or otherwise attached to the container), with multiple filter line ports of the manifold disposed within the interior cavity and at least one waste port disposed on the flexible exterior surface. In such embodiments, multiple filters within the interior cavity are fluidly connected to the multiple filter line ports via multiple filter lines, allowing waste to be extracted from the container via the at least one waste port. In embodiments, the manifold may be a stopcock manifold. As will be appreciated, in such embodiments, an external manifold is not required, and an external waste line is connected to the pump and waste bag.

[0069] In embodiments with multiple filters / filter membranes, the cumulative surface area of ​​the filters / filter membranes can be comparable to that of a single 7 inch by 7 inch or 15 inch by 15 inch filter.

[0070] In certain embodiments, filter selection and the flow of spent media / waste to the waste container can be automated or electronically controlled. For example, a flow sensor (not shown) located in the external waste line 374, 376, etc., can detect a clogged filter (or filter membrane) and send a warning to the user, automatically shut down the problematic filter, or select another filter within the vessel's internal cavity that has not yet been used (or is known to be unclogged). In embodiments, filter selection via a stopcock or manifold can be remotely controlled by the user via a smart device or other controller, or scheduled / preprogrammed. That is, filters can be activated in an alternating sequence to prevent clogging.

[0071] In this regard, embodiments of the present invention also contemplate a method for filtering waste materials in a rocker bioreactor. In one aspect, the method includes initiating a bioprocessing procedure, e.g., cell culture, in a vessel having an internal cavity with a plurality of filters. The method further includes determining whether one or more of the plurality of filters are clogged. If a filter is clogged, the method selects and activates an unclogged filter to allow waste materials to be extracted from the vessel. In certain embodiments, the method includes periodically selecting, activating, and deactivating different filters from the plurality of filters to prevent future filter clogs. The method may also include an initial step of activating all filters present in the vessel to maximize waste material removal.

[0072] As previously mentioned, in embodiments, the method steps may be automated, scheduled, or pre-programmed, for example, via multiple sensors and controllers. In other embodiments, a user may manually perform the method steps of the present invention.

[0073] As used herein, elements or steps described in the singular followed by the words "a" or "an" should be understood not to exclude a plurality of such elements or steps, unless expressly excluded. Furthermore, references to "one embodiment" or "embodiments" of the present invention are not intended to be interpreted as excluding the existence of additional embodiments that also incorporate the recited features. Furthermore, unless expressly stated to the contrary, embodiments "comprising," "including," or "having" an element or elements having a particular characteristic may include additional elements that do not have that characteristic. [Explanation of symbols]

[0074] 10 Rocker Bioreactor System 12 Bioreactor 14, 382, ​​446, 546, 646 Peristaltic pump 16 Rocker Bioreactor 20 trays 28 Lid 30 Bags 32 Cell culture media, fluid media 34, 134, 137, 234, 237, 334, 337, 370, 372, 631, 633, 635, 637 Disposal Ports 36, 40, 380, 384 tubes 38, 138, 238, 362, 364 filters 130, 230, 330, 360, 430, 530, 630 containers 136, 236, 336, 339 Disposal lines 139, 239, 374, 376, 432, 434, 436, 439, 532, 534, 536, 539, 632, 634, 636, 639 External waste line 140, 141, 240, 241 Filter Line 247, 431, 433, 435, 437, 531, 533, 535, 537 Disposal Ports 300, 400, 500, 600 Bioprocess Systems 335, 338, 420, 422, 424, 426, 520, 522, 524, 526, 620, 622, 624, 626 Filter membranes 340 First Filter Line 341 Second Filter Line 342, 378 Stopcock Manifold 348, 448, 548, 648 Disposal Bags 350 Section 366, 368 Internal filter line 442, 642 four-way manifold 552 Rigid Plate

Claims

1. a vessel having a flexible exterior defining an interior cavity configured to receive a fluid for a bioprocess, the vessel being configured to be selectively attached to a bioreactor; at least one filter within the interior cavity for retaining cells within the vessel while waste is extracted from the interior cavity; a plurality of waste ports formed on the flexible outer surface, each of the plurality of waste ports being fluidly connected to the at least one filter to allow waste to be extracted from the container; 1. An apparatus for bioprocessing comprising: The apparatus, wherein the vessel is configured to reduce the likelihood of a filter becoming clogged when cell densities are high during bioprocessing within the vessel.

2. 10. The apparatus of claim 1, wherein the bioreactor is a rocker bioreactor and the bioprocessing is a continuous perfusion cell culture process.

3. 10. The device of claim 1, further comprising a stopcock manifold fluidly connected to a plurality of waste ports via external waste lines, the stopcock manifold allowing a user to select an unclogged portion of the at least one filter when another portion of the at least one filter becomes clogged.

4. 10. The device of claim 1, wherein the at least one filter is a plurality of filters, and each of the plurality of waste ports is fluidly connected to a separate filter of the plurality of filters so as to enable waste to be extracted from the container.

5. 5. The device of claim 4, wherein the multiple filters are separate filter membranes formed on a single piece of material.

6. 5. The device of claim 4, further comprising a manifold fluidly connected to the plurality of waste ports via external waste lines, the manifold enabling waste to be simultaneously extracted through each of the plurality of filters.

7. 7. The apparatus of claim 6, wherein the manifold is a stopcock manifold that allows a user to select an unclogged filter from the plurality of filters in the event that another filter from the plurality of filters becomes clogged.

8. 5. The device of claim 4, wherein each of the plurality of filters is fluidly connected to the waste port via a filter line disposed within the internal cavity.

9. 5. The device of claim 4, wherein the plurality of filters is two filters, each filter fluidly connected to a separate waste port via a filter line.

10. 5. The device of claim 4, wherein the plurality of filters is four filters, each filter fluidly connected to a separate waste port via a filter line.

11. 10. The device of claim 1, wherein the at least one filter comprises pores having a pore size of about 1.2 μm.

12. 10. The device of claim 1, wherein the high cell density is greater than about 1e7 cells / mL.

13. The device of claim 1 , wherein a plurality of filters are secured within the interior cavity.

14. a vessel having a flexible exterior defining an interior cavity configured to receive a bioprocessing fluid, the vessel being configured to be selectively attached to a rocker bioreactor; a plurality of filters within the interior cavity for retaining cells within the vessel during a continuous perfusion cell culture process; a plurality of waste ports formed on the flexible exterior surface, the waste ports fluidly connected to the plurality of filters via filter lines disposed within the interior cavity to allow waste to be extracted from the container; a manifold fluidly connected to the plurality of waste ports via external waste lines; a waste bag fluidly connected to the manifold for receiving waste from the vessel during the continuous perfusion cell culture process; 1. A system for bioprocessing comprising: The manifold allows for simultaneous extraction of waste through each of the plurality of filters.

15. 15. The system of claim 14, wherein the manifold is a stopcock manifold that allows a user to select an unclogged filter from the plurality of filters if another filter from the plurality of filters becomes clogged due to high cell density during the continuous perfusion cell culture process.

16. 15. The system of claim 14, wherein the multiple filters are multiple separate filter membranes formed on a single piece of material.

17. 15. The system of claim 14, wherein the plurality of filters are two filters each fluidly connected to a separate waste port via a filter line.

18. 15. The system of claim 14, wherein the plurality of filters is four filters each fluidly connected to a separate waste port via a filter line.

19. 15. The system of claim 14, wherein each of the plurality of filters includes pores having a pore size of about 1.2 μm.

20. 16. The system of claim 15, wherein the high cell density is greater than about 1e7 cells / mL.

21. a vessel having a flexible exterior defining an interior cavity configured to receive a fluid for a bioprocess, the vessel being configured to be selectively attached to a bioreactor; a plurality of filters within the interior cavity for retaining cells within the vessel while waste is extracted from the interior cavity; a manifold disposed on the vessel, the manifold having a plurality of filter line ports within the interior cavity and at least one waste port on the flexible exterior surface; a plurality of filter lines fluidly connecting a plurality of filters to a plurality of filter line ports on the manifold so that waste can be extracted from the container through the at least one waste port; A bioprocessing vessel comprising: A bioprocessing vessel configured to reduce the likelihood of filter clogging when cell densities are high during bioprocessing within the vessel.

22. 22. The vessel of claim 21, wherein the manifold is a stopcock manifold that allows a user to select an unclogged filter from the plurality of filters if another filter from the plurality of filters becomes clogged under conditions of high cell density during bioprocessing.

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