Apparatus, system and method for filtering waste in shaker bioreactor

Abstract

An apparatus for biological treatment includes a container having a flexible exterior surface, the container defining an interior cavity configured to receive a fluid for biological treatment, the container configured for selective attachment to a bioreactor. The container comprises: at least one filter within the interior cavity for retaining cells in the container when waste is extracted from the interior cavity; and a plurality of waste ports formed on the flexible outer surface, each of the plurality of waste ports being fluidly connected to at least one filter to allow extraction of waste from the container. The container is configured to reduce the likelihood of filter clogging at high cell density during a biological treatment procedure in the container.

Description

Technical Field

[0001] Embodiments of the present invention generally relate to bioprocessing apparatus, systems and methods, and more particularly to filtering waste from a vessel of a shaker bioreactor during perfusion processing. Background Art

[0002] Bioreactors are often used to perform biochemical and / or biological processes and / or manipulate liquids and other products of such processes. Such bioreactors typically include a flexible or collapsible container, such as a single-use disposable bag supported by an external rigid structure. These "single-use" bioreactors include stirred tank reactors (where the sterile disposable bag and agitator are contained within a rigid tank), and shaker bioreactors (where the single-use bag is secured to a rocking platform).

[0003] In a shaker bioreactor, a disposable bag is secured to a tray, which in turn is attached to a motorized rocking device. The rocking device causes the tray to pivot back and forth around an axis / pivot point. The motion generated by the rocking device induces waves in the cell culture matrix within the bag. The waves provide mixing and gas-to-liquid transfer of oxygen supplied to the bag, resulting in an ideal environment for cell growth. To monitor and promote cell growth, the shaker bioreactor includes (among other features) a pump to add fresh cell growth / expansion matrix to the bag and remove used matrix therefrom.

[0004] In some known shaking bioreactors, removal of spent substrate is accomplished via a pump that draws the substrate out of the bag and into a waste receptacle via external tubing connected to a waste port on the bag wall. In perfusion processes, where fresh nutrients are continuously provided while spent substrate and waste products are removed, filters are used to retain cells in the bag during waste removal.

[0005] Reference Figure 3 , such filters (e.g., filter 38) are typically placed or floated on the top of the cell culture matrix 32 in the bag 30, and have its own pipeline 40 inside the bag, which is connected to the waste port. These filters include a series of small openings or holes on the side facing the cell matrix, which are sized to prevent cells from passing through the filter and outside the bag, while allowing the removal of spent matrix and other waste products. The opposite filter side includes a solid non-porous surface, and the internal pipeline 40 is typically formed or attached to this solid non-porous surface. In use, the pump applies suction to the filter 38 via the external line connected to the waste port, and waste is extracted from the bag 30, and the cells are retained in the holes of the filter.

[0006] However, with known bags, the potential for filter clogging exists at higher cell densities (e.g., those exceeding about 1e7 cells / mL). In particular, such filters may become clogged in and around the area where the internal tubing 40 is formed or attached to the filter 38. This clogging may exist even when other portions of the filter are not clogged, which results in the filter being potentially susceptible to clogging and having much less effect than its overall footprint.

[0007] As will be appreciated, such blockages can be detrimental to cell growth because it prevents spent matrix and other waste products from being removed from the bag. This is generally undesirable because biologics are time consuming to produce and have high value. In fact, this can be particularly severe in cell therapy situations where the cellular contents of the bag may be urgently needed.

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

[0009] Some embodiments commensurate in scope with the original claimed subject matter are summarized below. These embodiments are not intended to limit the scope of the claimed subject matter, but these embodiments are only intended to provide a brief overview of possible embodiments. In fact, the present disclosure may include various forms, which may be similar to or different from the embodiments set forth below.

[0010] According to one aspect of the invention, an apparatus for bioprocessing includes a container having a flexible outer surface, the container defining an interior cavity configured to receive a fluid for bioprocessing, the container configured for selective attachment to a bioreactor. The apparatus also includes: at least one filter within the interior cavity for retaining cells in the container while waste is extracted from the interior cavity; and a plurality of waste ports formed on the flexible outer surface, each of the plurality of waste ports being fluidly connected to at least one filter to allow waste to be extracted from the container. The container is configured to reduce the likelihood of filter clogging at high cell densities during a bioprocessing protocol in the container.

[0011] In embodiments, the bioreactor is a shaker bioreactor and the bioprocessing protocol is a continuous perfusion cell culture process.

[0012] In an embodiment, the apparatus further comprises a shutoff tap manifold fluidly connected to the plurality of waste ports via external waste lines, the shutoff tap manifold allowing a user to select an unblocked portion of the at least one filter when another portion of the at least one filter becomes blocked.

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

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

[0015] In an embodiment, the apparatus 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 an embodiment, the manifold may be a shutoff tap manifold that allows a user to select an unclogged filter from the plurality of filters when another filter from the plurality of filters becomes clogged.

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

[0018] In an embodiment, the plurality of filters may be two filters, each of which is fluidly connected to a separate waste port via a filter line.

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

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

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

[0022] In an embodiment, a plurality of filters are secured within the interior cavity.

[0023] According to another aspect of the present invention, a system for bioprocessing includes: a container having a flexible outer surface, the container defining an internal cavity configured to receive a fluid for bioprocessing, the container configured for selective attachment to a shaker bioreactor; and a plurality of filters within the internal cavity for retaining cells in the container during a continuous perfusion cell culture process. The system also includes: a plurality of waste ports formed on the flexible outer surface, the plurality of waste ports being fluidly connected to the plurality of filters via filter lines positioned within the internal cavity to allow waste to be extracted from the container; and a manifold fluidly connected to the plurality of waste ports via an external waste line. The system also includes a waste bag, the waste bag being fluidly connected to the manifold for receiving waste from the container during a continuous perfusion cell culture process. The manifold allows waste to be extracted simultaneously through each of the plurality of filters.

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

[0025] In embodiments, the plurality of filters may be a plurality of individual filter membranes formed on a single piece of material.

[0026] In an embodiment, the plurality of filters may be two filters, each fluidly connected to a separate waste port via a filter line.

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

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

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

[0030] According to another aspect of the invention, a container for bioprocessing includes a container having a flexible outer surface, the container defining an internal cavity configured to receive a fluid for bioprocessing, the container being configured for selective attachment to a bioreactor. The container also includes: a plurality of filters within the internal cavity for retaining cells in the container while extracting waste from the internal cavity; and a manifold positioned on the container having a plurality of filter line ports within the internal cavity and at least one waste port on the flexible outer surface. The container further includes a plurality of filter lines that fluidly connect the plurality of filters to the plurality of filter line ports on the manifold to allow waste to be extracted from the container through at least one waste port. The container is configured to reduce the likelihood of filter clogging at high cell densities during a bioprocessing protocol in the container.

[0031] In embodiments, the manifold may be a shutoff stopcock manifold that allows a user to select an unclogged filter from a plurality of filters if another filter in the plurality of filters becomes clogged at high cell density during a bioprocessing procedure. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0033] Figure 1 is a perspective view of a shaker bioreactor system suitable for use with embodiments of the present invention;

[0034] Figure 2 It is a combination Figure 1A perspective view of a known bioreactor vessel used in a shaker bioreactor depicting a filter of the vessel;

[0035] Figure 3 yes Figure 2 A side view of a bioreactor container / bag;

[0036] Figure 4 is a perspective view of a bioreactor container and a filter according to an embodiment of the present invention;

[0037] Figure 5 is a perspective view of a bioreactor vessel and filter according to an alternative embodiment of the present invention;

[0038] Figure 6 is a perspective view of a bioreactor container and a filter according to another embodiment of the present invention;

[0039] Figure 7 is a schematic diagram of a bioreactor vessel, filter, and related equipment according to an embodiment of the present invention;

[0040] Figure 8 is a schematic diagram of a bioreactor vessel, filters, and associated equipment depicting a four filter and port configuration according to an embodiment of the present invention;

[0041] Fig. 9 According to an embodiment of the present invention Figure 8 a schematic diagram of a filter and port configuration of FIG. 1 , wherein the filter is mounted on a plate anchored to a bottom container surface; and

[0042] Fig.10 According to another embodiment of the present invention Figure 8 Schematic diagram of a filter and port configuration in which the filter is tethered to the bottom container surface. DETAILED DESCRIPTION

[0043] Reference will now be made in detail to exemplary embodiments of the present invention, examples of which are illustrated in the accompanying drawings. Whenever possible, the same reference symbols are used throughout the drawings to refer to the same or like parts.

[0044] As used herein, the terms "flexible" or "contractible" refer to structures or materials that are pliable or capable of bending without breaking, and may also refer to compressible or expandable materials. An example of a flexible structure is a bag formed from a polyethylene film. The terms "rigid" and / or "semi-rigid" are used interchangeably herein to describe "non-contractible" structures, in other words, structures that do not fold, contract, or otherwise deform to significantly reduce their extended dimensions under normal forces.

[0045] "Container" as the term is used herein means a flexible bag, a flexible vessel, a semi-rigid vessel, or a rigid vessel, as the case may be. The term "container" as used herein is intended to include bioreactor containers having flexible or semi-rigid walls or portions of walls, single-use 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, matrix / buffer preparation systems, and filtration / purification systems.

[0046] As used herein, the term "bag" means a flexible or semi-rigid vessel or container that is used, for example, as a bioreactor or mixer for the contents therein. Although embodiments of the present invention are described as being used for use with bioprocessing bags (including but not limited to bioreactor bags and mixer bags), embodiments may also be configured for use with other bags or containers.

[0047] Embodiments can be used to perform a variety of biological processes in a shaker bioreactor, and are not limited to cell expansion. Certain embodiments can be used broadly in bioprocessing and biochemical environments, and potentially in non-biological / biochemical contexts. Likewise, while the embodiments are described and depicted with respect to a specific shaker bioreactor system and in conjunction with a continuous perfusion cell culture process, the embodiments can potentially be used with other types of reactors / mixers, particularly those with containers in which continuous cell culture is performed and / or waste is removed via filters.

[0048] Likewise, embodiments are not limited to any particular size / shape of container / bag. Although embodiments may be particularly suitable and / or described for use with 20L-50L flexible bags, other sizes may be used, for example, 2L-10L.

[0049] Reference Figure 1 , depicts an exemplary shaker bioreactor system 10 suitable for use with embodiments of the present invention. As shown, the system 10 includes a bioreactor 12 connected to one or more peristaltic pumps 14. The bioreactor 12 is also operatively connected via a wired or wireless connection (not shown) to a controller, e.g., a local or remote computer that provides process / protocol monitoring, etc.

[0050] The bioreactor 12 includes a removable tray 20 configured to selectively receive and support a container / bag. In an embodiment, the tray 20 includes a removable lid 28 that features a hinged door 30 that can be raised to gain access to the tray 20 and any container secured thereto. The bioreactor 12 also includes a base 16 that is operatively connected to the tray 20. The base 16 houses a pivot mechanism, such as a motor, that enables the tray 20 to pivot back and forth.

[0051] In use, the container / bag is secured to the tray 20 via one or more attachment mechanisms, such as selectively lockable clamps positioned at opposite ends of the tray 20. The tray 20 and bag are then pivoted back and forth, creating a rocking motion that induces waves in the cell culture matrix within the bag to promote growth.

[0052] Now refer to Figure 2 and Figure 3 , depicting a conventional flexible reactor container / bag 30 with a fluid 32 containing a growth medium. Removal of the spent medium is accomplished via a pump 14 (e.g., a peristaltic pump) which draws the medium out of the bag 30 and into a waste receptacle via an external line / tubing 36 connected to a waste port 34 on the bag wall. To provide perfusion, e.g., to retain cells in the bag 30 while the spent fluid medium 32 is removed, a filter 38 is positioned within the bag 30. The filter 38 has its own filter line / tubing 40 inside the bag which is also connected to the waste port 34. However, with known filters, there are problems with the ability to filter cells at high cell densities (e.g., greater than about 1e7 cells / mL (1×10 7 ) of the cell density).

[0053] In particular, the filter 38 may become clogged in and around the area where the internal tubing 40 is formed or attached to the filter 38. This clog may exist even when other portions of the filter 38 are not clogged, which causes the filter 38 to potentially be susceptible to clogging and less effective than its overall footprint. As will be appreciated, such clogs may be detrimental to cell growth because it prevents spent matrix and other waste products from being removed from the bag 30.

[0054] Now refer to Figure 4 , depicting an apparatus for bioprocessing according to an embodiment of the present invention. As shown, the apparatus includes a bag / container 130 having a flexible outer surface that defines an internal cavity configured to receive a fluid 132 (e.g., a cell culture matrix, etc.) for bioprocessing. The container 130 is configured for selective attachment to the tray 20 of the shaker bioreactor 16.

[0055] 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 depicted embodiment, the filter 138 extends transversely relative to the bag 30 and is connected to a plurality of internal tubing / filter lines 140, 141 and to a plurality of waste ports 134, 137 to which a waste bag (not shown) is fluidly connected via external waste lines 136, 139. As will be appreciated, the filter lines 140, 141 are fluidly connected to the filter 138 to allow waste that has passed through the holes on the side of the filter facing the cell matrix (not shown) to exit the container 130 via the waste ports 134, 137.

[0056] Waste ports 134, 137 are formed on a flexible exterior surface of the container 130. As will be appreciated, the waste ports 134, 137 may be welded to the container 130, or may be molded into or otherwise integral with the container 130. However, embodiments are not limited to a particular size, shape, or configuration of waste ports 134, 137, and conventional ports may be suitable so that 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 proximate to where the filter lines 140, 141 connect to at least one filter 138 (e.g., directly above it).

[0057] The filter 138 has a side facing the cell culture substrate that includes a plurality of holes (not shown). The filter 138 also has an opposite side that is solid and non-porous, to which the filter lines 140, 141 are attached or formed. In an embodiment, the filter 138 is a laminate or composite of a porous material and a non-porous material with a mesh spacer in the middle to prevent the porous material and the non-porous material from shrinking on each other when suction is applied. The filter lines 140, 141 can be fixed to the filter by a variety of methods (such as heat welding or molding).

[0058] As will be appreciated, embodiments are not limited to any particular filter material, construction, or pore size, but in certain embodiments, the filter may be made of a polymer such as polyvinylidene fluoride (PVDF) or polysulfone (PES), and may have a pore size of approximately 1.2 μm. Likewise, the filter wires 140, 141 are not limited to a particular polymer material or inner or outer diameter, length, or shape.

[0059] In certain embodiments, the filter 138 may be connected to more than two filter lines. For example, the filter 138 may be connected to a filter line positioned at each corner of the filter 138 or in an array or other arrangement along the filter 138. In fact, while embodiments use at least two filter lines, they are not limited to only two, and the number of filter lines may be determined by a variety of factors, including the amount of space for corresponding waste ports on the flexible exterior of the bag. In a particular embodiment, four filter lines (and four waste ports) may be employed (see Figure 8-10 ).

[0060] Similarly, embodiments are not limited to a particular shape or size of filter, but quadrilateral filters are conventional. As will be appreciated, other filter shapes may be used without departing from the invention. In embodiments, filter 138 may be approximately 7"x7" (17.8 cm x 17.8 cm) for 2L and 10L containers / bags, and may be approximately 15"x15" (38.1 cm x 38.1 cm) for 20L and 50L containers / bags, but other sizes may be used.

[0061] The container / bag itself can be made of a variety of materials, and the present invention is not limited in this respect. In an embodiment, the container can be a multilayer laminate of USP Class VI material. The container can be completely or partially transparent to allow viewing of the internal cavity.

[0062] Refer again Figure 4 , the filter lines 140, 141 reduce the likelihood of clogging by providing suction at multiple locations on the filter 138. In this way, a larger portion of the filter 138 can be effectively used. In addition, as described in more detail below, in some embodiments, one or more of the filter lines 140, 141 can be selectively activated / deactivated via, for example, a shutoff manifold, so when another portion becomes clogged, the user can select an unclogged portion of the filter for use.

[0063] Now go to Figure 5 , an embodiment featuring a longitudinally extending filter 238 may also be used. This embodiment is similar to Figure 4 That is, the device includes a container 230 with a filter 238 in the interior cavity. The filter 238 is fluidly connected to waste ports 234, 237 via filter lines 240, 241, respectively. Waste lines 236, 239 extend from the waste ports 234, 237 to a waste bag (not shown).

[0064] Now refer to Figure 6, another embodiment of the present invention is described. In this embodiment, container 330 has a filter, which includes multiple filters, which are separate filter membranes 335, 338 formed on a single piece or sheet material (e.g., polymer laminate). As used herein, the term "filter" includes such filter membranes except physically separated filters. Each filter membrane 335, 338 has a hole on its side (not shown) facing the cell culture matrix. Filter membranes 335, 338 are separated by sections 350, which effectively isolate or seal each of the filter membranes 335, 338 from another. This can be achieved by a variety of methods, and in an embodiment, section 350 is a heat seal between filter membranes 335, 338.

[0065] 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 and second filter lines 340, 341 use separate (e.g., first and second) waste lines 336, 339 that are operatively connected to a pump and waste bag.

[0066] In this embodiment (and Figure 7 In the embodiment depicted in ), it is contemplated that the filter membranes 335, 338 can be used independently of each other via a shutoff plug manifold or other switching device. In this aspect, when another filter from a plurality of filters becomes clogged, a user can select a filter from the plurality of filters that is not clogged.

[0067] As will be appreciated, in some embodiments, multiple filters may be attached to each other (or to specific filters in the multiple filters) by other structures or mechanisms. That is, they need not be filter membranes formed from a single piece of material, but may be completely separate filters welded together at certain locations.

[0068] Now refer to Figure 7 , the system 300 for bioprocessing includes a container 360 with a flexible outer surface, which defines an internal cavity. The internal cavity includes 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 material.

[0069] The filters 362, 364 are fluidly connected to a plurality of 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 and 376 are connected to the waste ports 370, 372 and to a three-way valve / stopcock manifold 378, which in turn is connected to a peristaltic pump 382 and waste bag 348 via lines / tubings 380, 384. As mentioned, this embodiment allows for the exchange of a clogged filter with an unused filter via the stopcock manifold 342 during a bioprocessing procedure.

[0070] Now refer to Figure 8 In a particular embodiment, a system 400 for bioprocessing includes a container 430 with a flexible outer surface that defines an interior cavity that includes a plurality of filters, such as four filters, such as filter membranes 420, 422, 424, 426, formed on a single piece of material (e.g., a polymer laminate). Figure 6 As in the embodiment of the invention, the filter membranes 420, 422, 424, 426 are isolated from each other by heat seals or other structures schematically represented by the depicted mesh. Much like the other embodiments, the filter membranes 420, 422, 424, 426 have a side facing the fluid containing holes that contacts the cell matrix when the filter floats on the cell matrix.

[0071] In this embodiment, each filter membrane 420, 422, 424, 426 includes a filter line inside the container, which is not depicted, but is configured according to the previously described embodiments. The filter lines (one filter line for each filter membrane 420, 422, 424, 426) connect the membrane fluid to multiple 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 waste ports 431, 433, 435, 437 and to a four-way manifold 442, which in turn is connected to a peristaltic pump 446 and a waste bag 448 via lines / tubing. As will be appreciated, this embodiment allows waste to be extracted from all four filter membranes simultaneously via manifold 442.

[0072] In an embodiment, manifold 442 is a stopcock manifold (or other switching manifold or device) that allows a plugged filter membrane to be exchanged with an unused or known non-plugged filter membrane during a bioprocessing procedure.

[0073] Now refer to Fig. 9In this embodiment, a system 500 for bioprocessing includes a container 530 with a flexible outer surface that defines an interior cavity that includes a plurality of filters, which are four filter membranes 520, 522, 524, 526 formed on a single piece of material (e.g., a polymer laminate). Figure 8 As in the embodiment of the invention, the filter membranes 520, 522, 524, 526 are isolated from each other by heat seals or other structures schematically represented by the depicted mesh. Again, the filter membranes 520, 522, 524, 526 have a side facing the fluid containing holes, which contacts the cell matrix when the filter floats on the cell matrix.

[0074] Each filter membrane 520, 522, 524, 526 includes a filter line that connects the membrane fluid to a plurality of 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 via lines / tubing to a waste bag 548. However, with respect to this embodiment, the filter membranes 520, 522, 524, 526 are secured to a rigid plate 552 that is anchored to the bottom surface of the container.

[0075] Similarly, if Fig.10 As shown in FIG. 6 , the bioprocessing system 600 includes a vessel 630 including four filter membranes 620, 622, 624, 626 formed on a single piece of material and including filter lines connecting the membrane fluids to a plurality of waste ports (e.g., four waste ports 631, 633, 635, 637), external waste lines 632, 634, 636, 639, a four-way manifold 642 (e.g., a stopcock manifold), a pump 646, and via lines / tubing to a waste bag 648. However, in this embodiment, although there is no Fig. 9 620, 622, 624, 626 are secured to the bottom surface of the container via a plurality of tethers. The tethers may be formed from a single piece of material as the filter membranes and / or may be welded to the bottom surface of the container. Of course, other attachment mechanisms may be employed without departing from the scope of the present invention.

[0076] Although the embodiments are described for use in conjunction with a manifold and a shutoff manifold, a variety of manifolds or switching mechanisms / valve groups may be used in general. These include pneumatic valves, etc. In embodiments, such manifolds / switching mechanisms may be operatively connected to a controller.

[0077] In certain embodiments, the container may have a manifold positioned on the container itself. In such embodiments, the manifold may be positioned on the flexible outer surface of the container (e.g., formed therein or otherwise attached thereto) such that a plurality of filter line ports of the manifold will be positioned within the internal cavity and at least one waste port will be positioned on the flexible outer surface. In such embodiments, a plurality of filters within the internal cavity will be fluidly connected to a plurality of filter line ports via a plurality of filter lines to allow waste to be extracted from the container through at least one waste port. In embodiments, the manifold may be a stopcock manifold. As will be appreciated, in such embodiments, an external manifold will not be required and an external waste line will be connected to a pump and a waste bag.

[0078] In embodiments featuring multiple filters / filter membranes, the cumulative surface area of ​​the filters / filter membranes may total to the cumulative surface area of ​​a single 7"x7" or 15"x15" filter.

[0079] In certain embodiments, the selection of filter and the flow of used substrate / waste in the waste receiver can be automated or electronically controlled in addition.For example, the flow sensor (not shown) positioned at, for example, on the external waste line 374,376 can detect the filter (or filter membrane) blocked, and can send an alarm to the user and / or automatically cut off the affected filter and select another (or known not blocked) filter to be used from those filters in the inner cavity of the container.In an embodiment, the selection of filter via cut-off cock or manifold can be operated remotely by the user via intelligent device or other controllers, or can be planned / preprogrammed.That is, the filter can prevent blocking with alternating sequential activation.

[0080] In this regard, embodiments of the present invention also contemplate methods for filtering waste in a shaking bioreactor. On the one hand, the method includes the step of starting a bioprocessing procedure (e.g., cell culture) in a container having an internal cavity with multiple filters. The method also includes determining whether one or more of the multiple filters are clogged. If the filter is clogged, the method includes selecting and activating a filter that is not clogged so that waste can be extracted from the container. In certain embodiments, the method also includes periodically selecting, activating, and deactivating different filters from the multiple filters to prevent future filter clogging. The method may also include an initial step of activating all filters present in the container to maximize the removal of waste.

[0081] As mentioned, in embodiments, the method steps may be automated, scheduled or pre-programmed, for example via a plurality of sensors and a controller. In other embodiments, a user may manually perform the method steps of the invention.

[0082] As used herein, elements or steps stated in the singular and beginning with the word "a" or "an" should be understood as not excluding plural said elements or steps, unless such exclusion is explicitly stated. In addition, reference to "an embodiment" or "an embodiment" of the present invention is not intended to be interpreted as excluding the existence of additional embodiments that also incorporate the stated features. In addition, unless explicitly stated to the contrary, embodiments that "comprise," "include," or "have" an element or elements having a particular property may include additional such elements that do not have that property.

Claims

1. A device for biological treatment, comprising: a container having a flexible exterior surface, the container defining an interior cavity configured to receive a fluid for bioprocessing, the container configured for selective attachment to a bioreactor; at least one filter within the interior cavity, the at least one filter being configured to retain cells in the container while waste is extracted from the interior cavity; as well as 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 extraction of waste from the container; Wherein the container is configured to reduce the likelihood of filter clogging at high cell densities during bioprocessing procedures in the container.

2. The device according to claim 1, wherein The bioreactor is a shaker bioreactor and the bioprocessing protocol is a continuous perfusion cell culture process.

3. The device according to claim 1, further comprising: A stopcock manifold is fluidly connected to the plurality of waste ports via external waste lines, the stopcock manifold allowing a user to select an unobstructed portion of the at least one filter when another portion of the at least one filter becomes obstructed.

4. The device according to 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 to allow extraction of waste from the container.

5. The device according to claim 4, wherein: The plurality of filters are individual filter membranes formed on a single piece of material.

6. The device according to claim 4, further comprising: A manifold is fluidly connected to the plurality of waste ports via external waste lines, the manifold allowing waste to be extracted simultaneously through each of the plurality of filters.

7. The device according to claim 6, wherein: The manifold is a shutoff tap manifold that allows a user to select an unclogged filter from the plurality of filters when another filter from the plurality of filters becomes clogged.

8. The device according to claim 4, wherein: Each of the plurality of filters is fluidly connected to a waste port via a filter line positioned in the interior cavity.

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

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

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

12. The device according to claim 1, wherein: The high cell density is greater than about 1e7 cells / mL.

13. The apparatus according to claim 1, wherein: A plurality of filters are secured within the interior cavity.

14. A system for bioprocessing, the system comprising: a container having a flexible exterior surface, the container defining an interior cavity configured to receive a fluid for bioprocessing, the container configured for selective attachment to a shaker bioreactor; a plurality of filters within the interior cavity, the plurality of filters being used to retain cells in the container during a continuous perfusion cell culture process; a plurality of waste ports formed on the flexible exterior surface, the plurality of waste ports being fluidly connected to the plurality of filters via filter lines positioned within the interior cavity to allow extraction of waste from the container; a manifold fluidly connected to the plurality of waste ports via external waste lines; as well as a waste bag fluidly connected to the manifold for receiving waste from the container during the continuous perfusion cell culture process; wherein the manifold allows for simultaneous extraction of waste through each of the plurality of filters.

15. The system of claim 14, wherein: The manifold is a shutoff 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 at high cell density during the continuous perfusion cell culture process.

16. The system of claim 14, wherein: The plurality of filters are a plurality of individual filter membranes formed on a single piece of material.

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

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

19. The system of claim 14, wherein: Each of the plurality of filters includes pores having a pore size of approximately 1.2 μm.

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

21. A container for bioprocessing, the container comprising: a container having a flexible exterior surface, the container defining an interior cavity configured to receive a fluid for bioprocessing, the container configured for selective attachment to a bioreactor; a plurality of filters within the interior cavity, the plurality of filters being used to retain cells in the container while waste is extracted from the interior cavity; a manifold positioned on the container, the manifold having a plurality of filter line ports within the interior cavity and at least one waste port on the flexible exterior surface; as well as a plurality of filter lines fluidly connecting the plurality of filters with the plurality of filter line ports on the manifold to allow extraction of waste from the container through the at least one waste port; Wherein the container is configured to reduce the likelihood of filter clogging at high cell densities during bioprocessing procedures in the container.

22. The container according to claim 21, wherein The manifold is a shutoff stopcock manifold that allows a user to select an unplugged filter from the plurality of filters if another filter from the plurality of filters becomes plugged at high cell density during the bioprocessing procedure.

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