Note: Within nine months of the publication of the mention of the grant of the European patent in the European PatentBulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with theImplementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has beenpaid. (Art. 99(1) European Patent Convention).

Printed by Jouve, 75001 PARIS (FR)

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(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention of the grant of the patent: 13.09.2017 Bulletin 2017/37

(21) Application number: 07872150.3

(22) Date of filing: 15.06.2007

(51) Int Cl.:B01F 13/08 (2006.01) C12M 1/06 (2006.01)

B01F 15/00 (2006.01)

(86) International application number: PCT/US2007/014050

(87) International publication number: WO 2008/088371 (24.07.2008 Gazette 2008/30)

(54) GAS DELIVERY CONFIGURATIONS, FOAM CONTROL SYSTEMS, AND BAG MOLDING METHODS AND ARTICLES FOR COLLAPSIBLE BAG VESSELS AND BIOREACTORS

GASZUFUHRKONFIGURATIONEN, SCHAUMSTOFFSTEUERSYSTEME UND BEUTELFORMUNGSVERFAHREN SOWIE ARTIKEL FÜR FALTBARE BEUTELGEFÄSSE UND BIOREAKTOREN

CONFIGURATIONS DE DISTRIBUTION DE GAZ, SYSTÈMES DE COMMANDE DE MOUSSE, ET PROCÉDÉ DE MOULAGE AU SAC ET ARTICLES POUR RÉCIPIENTS ET BIORÉACTEURS DE TYPE SACS RÉTRACTABLES

(84) Designated Contracting States: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR

(30) Priority: 16.06.2006 US 814647 P

(43) Date of publication of application: 08.04.2009 Bulletin 2009/15

(73) Proprietor: GE Healthcare Bio-Sciences Corp.Marlborough, MA 01752 (US)

(72) Inventors: • GALLIHER, Parrish, M.

Littleton, Massachussets 01640 (US)• HODGE, Geoffrey, L.

Sutton, MA 01590 (US)• FISHER, Michael

Ashland, MA 01721 (US)

(74) Representative: Larsson, Jan Anders et alGE Healthcare Bio-Sciences AB Björkgatan 30751 84 Uppsala (SE)

(56) References cited: EP-A- 0 358 053 EP-A- 0 399 972EP-A- 1 329 301 EP-A2- 0 530 820WO-A-92/08743 WO-A-2005/118771DD-A1- 278 505 DE-A- 2 548 441GB-A- 1 181 877 GB-A- 2 342 418JP-A- 06 153 902 JP-A- 63 016 009US-A- 2 985 916 US-A- 4 209 259US-A- 4 310 437 US-A- 4 355 906US-A- 4 727 040 US-A- 4 883 759US-A- 4 910 054 US-A- 4 943 404US-A- 4 987 082 US-A- 5 075 234US-A- 5 935 843 US-A1- 2002 076 815US-A1- 2005 077 239 US-A1- 2006 033 222US-A1- 2006 129 026 US-B1- 6 581 802

• DATABASE WPI Week 200537 Thomson Scientific, London, GB; AN 2005-357148 XP002493795 & CN 1 581 708 A (LG ELECTRONIC RES & DEV CENT CO LTD) 16 February 2005 (2005-02-16)

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Description

FIELD OF INVENTION

[0001] The present invention relates generally to sys-tems for containing and manipulating fluids, and in certainembodiments, to systems and methods involving col-lapsible bags that may be used as reactors for performingchemical, biochemical and/or biological reactions con-tained therein.

BACKGROUND

[0002] A variety of vessels for manipulating fluidsand/or for carrying out chemical, biochemical and/or bi-ological reactions are available. For instance, biologicalmaterials (e.g., animal and plant cells) including, for ex-ample, mammalian, plant or insect cells and microbialcultures can be processed using bioreactors. Traditionalbioreactors, which are typically designed as stationarypressurized vessels, or disposable bioreactors, many ofwhich utilize plastic sterile bags, may be used. For in-stance, US4310437A and DE2548441A disclose biore-actors comprising rotating antifoaming devices. JP H06153902A discloses a bioreactor comprising a flexible cul-ture bag.Although reaction systems and other fluid manipulatingsystems (e.g., mixing systems) are known, improve-ments to such systems would be beneficial.

SUMMARY OF THE INVENTION

[0003] The present invention relates to a vessel con-figured to contain a volume of liquid comprising a con-tainer able to contain the volume of liquid, wherein thecontainer is a collapsible bag; a reusable support struc-ture surrounding and containing the collapsible bag; anda magnetically-driven antifoaming device, which is posi-tioned in a head space of the container when the con-tainer contains the volume of liquid, the antifoaming de-vice being configured and arranged to break up foam inthe head space during rotation of at least a portion of theantifoaming device, wherein the antifoaming device com-prises an impeller that is magnetically rotated about asingle axis.The present disclosure relates generally to systems forcontaining and manipulating fluids, and in certain em-bodiments, to systems and methods involving supportedcollapsible bags that may be used as bioreactors for per-forming chemical, biochemical and/or biological reac-tions contained therein. The subject matter of the presentdisclosure involves, in some cases, interrelated prod-ucts, alternative solutions to a particular problem, and/ora plurality of different uses of one or more systems and/orarticles.[0004] In one aspect of the disclosure, a series of ves-sels are provided. In one embodiment, a vessel config-ured to contain a volume of liquid is provided. The vessel

comprises a collapsible bag for containing the volume ofliquid, the collapsible bag having a volume of at least 2liters and a support structure surrounding and containingthe collapsible bag. The vessel also includes a first sparg-er connected to the collapsible bag, the first sparger beingin fluid communication with a source of a first gas com-position, and a second sparger connected to the collaps-ible bag, the second sparger being in fluid communicationwith a source of a second gas composition different fromthe first gas composition.[0005] In another embodiment, a vessel configured tocontain a volume of liquid comprises a collapsible bagable to contain the volume of liquid and a support struc-ture surrounding and containing the collapsible bag. Thevessel also includes a first sparger connected to the col-lapsible bag, the first sparger having a first aperture size,wherein at least a portion of the first sparger is dimen-sioned to be connected to a source of a first gas, and asecond sparger connected to the collapsible bag, thesecond sparger having a second aperture size, whereinat least a portion of the second sparger is dimensionedto be connected to a source of a second gas.[0006] In another embodiment, a vessel configured tocontain a volume of liquid comprises a container able tocontain the volume of liquid. The vessel may also includea magnetically-driven antifoaming device, at least a por-tion of which is positioned in a head space of the containerwhen the container contains the volume of liquid. Theantifoaming device is configured and arranged to breakup foam in the head space during rotation of at least aportion of the antifoaming device.[0007] In another embodiment, a vessel configured tocontain a volume of liquid comprises a collapsible bagable to contain the liquid and a reusable support structuresurrounding and containing the collapsible bag. The ves-sel also comprises a pressure sensor for determining apressure in the collapsible bag, the pressure sensor influid communication with the collapsible bag, and an an-tifoaming device associated with the collapsible bag andconfigured to break up foam in the collapsible bag. Thevessel may also include a control system operatively as-sociated with the pressure sensor and the antifoamingdevice, wherein the control system regulates the anti-foaming device upon receipt of a signal from the pressuresensor.[0008] In another embodiment, a collapsible bag con-figured to contain a volume of at least 2 liters is provided.The collapsible bag comprises a first rotatable impellerpositioned at a bottom portion of the collapsible bag, thefirst impeller able to be magnetically rotated, and a sec-ond impeller positioned at a top portion of the collapsiblebag, the second impeller able to be magnetically rotated.[0009] In another embodiment, a container able to con-tain a volume of liquid is provided. The container com-prises a collapsible bag having a volume of at least 40liters, wherein the collapsible bag does not include anyseams joining two or more flexible wall portions of thecollapsible bag, and a reusable support structure sur-

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rounding and containing the collapsible bag.[0010] In another aspect of the disclosure, a series ofmethods are provided. In one embodiment, a methodcomprises positioning a rigid component in a mold havinga shape configured to mold a container having a volumeof at least 10 mL, and introducing a first polymer or pol-ymer precursor into the mold. The method includes form-ing a seamless container within the mold by solidifyingthe polymer or polymer precursor to form the container,wherein the component is incorporated in the container.[0011] In another embodiment, a method comprisesintroducing a first polymer or polymer precursor into amold, the mold having a shape configured to mold a con-tainer having a volume of at least 10 mL, the mold furthercomprising at least one mandrel for forming a functionalcomponent of a liquid containment system. The methodalso includes forming a container within the mold, forminga component within the mold utilizing the mandrel, andjoining the functional component and the container with-out welding.[0012] In another embodiment, a method comprisesintroducing a first polymer or polymer precursor into amold, the mold having a shape configured to mold a col-lapsible bag having a volume of at least 10 mL and alsoconfigured to mold a base including a shaft configuredto support a magnetic impeller and forming a collapsiblebag within the mold. The method also includes introduc-ing a second polymer precursor into the mold, forming acomponent of the mixing system by solidifying the secondpolymer precursor, and joining the component of the mix-ing system and the collapsible bag without welding.[0013] In another embodiment, an article comprises acollapsible bag comprising a flexible wall portion and arigid portion comprising a base including a shaft config-ured to support a magnetic impeller, wherein the rigidportion is embedded with the flexible wall portion.[0014] Other advantages and novel features of thepresent invention will become apparent from the follow-ing detailed description of various non-limiting embodi-ments of the invention when considered in conjunctionwith the accompanying figures. In cases where thepresent specification and a document incorporated byreference include conflicting and/or inconsistent disclo-sure, the present specification shall control. If two or moredocuments incorporated by reference include conflictingand/or inconsistent disclosure with respect to each other,then the document having the later effective date shallcontrol.

BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Non-limiting embodiments of the present inven-tion will be described by way of example with referenceto the accompanying figures, which are schematic andare not intended to be drawn to scale. In the figures, eachidentical or nearly identical component illustrated is typ-ically represented by a single numeral. For purposes ofclarity, not every component is labeled in every figure,

nor is every component of each embodiment of the in-vention shown where illustration is not necessary to allowthose of ordinary skill in the art to understand the inven-tion. In the figures:

Fig. 1 illustrates one embodiment of the invention,showing a container contained within a supportstructure;Figs. 2A-2C illustrate techniques for forming a seam-less container, according to another embodiment ofthe invention;Fig. 3 illustrates a vessel for carrying out fluid ma-nipulations including biological, chemical, and bio-chemical processes, according to another embodi-ment of the invention;Figs. 4A-4B illustrate various devices including im-pellers, according to another embodiment of the in-vention;Fig. 5 shows an impeller magnetically coupled to anexternal motor, according to another embodiment ofthe invention;Fig. 6 shows an impeller, according to another em-bodiment of the invention;Fig. 7 shows an example of an antifoaming system,according to another embodiment of the invention;Fig. 8 shows another example of an antifoaming sys-tem, according to another embodiment of the inven-tion; andFig. 9 shows an example of a control and feedbackprocess, according to another embodiment of theinvention.

DETAILED DESCRIPTION

[0016] The present invention relates generally to sys-tems for containing and manipulating fluids, and in certainembodiments, to systems and methods involving col-lapsible bags that may be used as reactors for performingchemical, biochemical and/or biological reactions con-tained therein. Generally, the invention provides a seriesof improvements and features for fluid containment sys-tems such as gas delivery configurations, foam controlsystems and bag molding methods and articles for sup-ported collapsible bag vessels and bioreactors. For in-stance, in one aspect, fluids contained in a vessel canbe sparged, e.g., such that a fluid is directed into a con-tainer of the vessel, and in some cases, the sparging canbe controlled by rapidly activating or altering the degreeof sparging as needed. Multiple spargers may be usedin some cases. In another aspect, the vessel includes aseamless collapsible bag. In some cases, the collapsiblebag may be injected, blown, or spin cast molded. In yetanother aspect, the vessel includes a device which canreduce the foam produced or contained within the vessel.Sensors and/or controllers may optionally be used tomonitor and/or control foaming.[0017] The following documents form part of the stateof the art: U.S. Provisional Patent Application Serial No.

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60/903,977, filed February 28, 2007, entitled "WeightMeasurements of Liquids in Flexible Containers," by P.A.Mitchell, et al. published as U.S. Patent Application Pub-lication No. 2008/0213874 on September 4, 2008; U.S.Patent Application Serial No. 11/147,124, filed June 6,2005, entitled "Disposable Bioreactor Systems andMethods," by G. Hodge, et al., published as U.S. PatentApplication Publication No. 2005/0272146 on December8, 2005; International Patent Application No.PCT/US2005/020083, filed June 6, 2005, entitled "Dis-posable Bioreactor Systems and Methods," by G. Hodge,et al., published as WO 2005/118771 on December 15,2005; and International Patent Application No.PCT/LTS2005/002985, filed February 3, 2005, entitled"System and Method for Manufacturing," by G. Hodge,et al., published as WO 2005/076093 on August 18,2005.[0018] Although much of the description herein in-volves an exemplary application of the present inventionrelated to bioreactors (and/or biochemical and chemicalreaction systems), the invention and its uses are not solimited, and it should be understood that aspects of theinvention can also be used in other settings, includingthose involving containment systems in general, as wellas systems for containment and/or processing of a fluidin a container (e.g., mixing systems). It should also beunderstood that while many examples provided hereininvolve the use of collapsible bags or flexible containers,aspects of the invention can be integrated with systemsinvolving non-collapsible bags, rigid containers, and oth-er configurations involving liquid containment.[0019] In one aspect, vessels configured to contain avolume of liquid are provided. In certain embodiments,the vessels are a part of a bioreactor system. For exam-ple, a non-limiting example of a bioreactor system includ-ing a container, such as a flexible container, is shown inthe schematic diagram of Fig. 1. As shown in the embod-iment illustrated in Fig. 1, vessel 10 includes a reusablesupport structure 14 (e.g., a stainless steel tank) that sur-rounds and contains a container 18. In some embodi-ments, the container is configured as a collapsible bag(e.g., a polymeric bag). Additionally and/or alternatively,all or portions of the collapsible bag or other containermay comprise a substantially rigid material such as arigid polymer, metal, and/or glass. In other embodiments,a rigid container is used in this configuration. The con-tainer may be disposable and may be configured to beeasily removable from the support structure. In some em-bodiments, the container is non-integrally connected tothe support structure. As used herein, the term "integrallyconnected," when referring to two or more objects,means separation of the two or more objects requirescausing damage to at least one of the object (or compo-nents of the object), for example, by breaking or peeling(e.g., separating components fastened together via ad-hesives, tools, etc.).[0020] If a collapsible bag is used, the collapsible bagmay be constructed and arranged for containing a liquid22, which may contain reactants, media, and/or other

components necessary for carrying out a desired processsuch as a chemical, biochemical and/or biological reac-tion. The collapsible bag may also be configured suchthat liquid 22 remains substantially in contact only withthe collapsible bag during use and not in contact withsupport structure 14. In such embodiments, the bag maybe disposable and used for a single reaction or a singleseries of reactions, after which the bag is discarded. Be-cause the liquid in the collapsible bag may not come intocontact with the support structure, the support structurecan be reused without cleaning. That is, after a reactiontakes place in container 18, the container can be removedfrom the support structure and replaced by a second(e.g., disposable) container. A second reaction can becarried out in the second container without having toclean either the first container or the reusable supportstructure.[0021] Also shown in Fig. 1 are an optional inlet port42 and optional outlet port 46, which can be formed inthe container and/or reusable support structure and canfacilitate more convenient introduction and removal of aliquid and/or gas from the container. The container mayhave any suitable number of inlet ports and any suitablenumber of outlet ports. For example, a plurality of inletports may be used to provide different gas compositions(e.g., via a plurality of spargers 47), and/or to allow sep-aration of gases prior to their introduction into the con-tainer. These ports may be positioned in any suitablelocation with respect to container 18. For instance, forcertain vessels including spargers, the container may in-clude one more gas inlet ports located at a bottom portionof the container. Tubing may be connected to the inletand/or outlet ports to form, e.g., delivery and harvestlines, respectively, for introducing and removing liquidfrom the container. Optionally, the container and/or sup-port structure may include a utility tower 50, which maybe provided to facilitate interconnection of one or moredevices internal to the container and/or support structurewith one or more pumps, controllers, and/or electronics(e.g., sensor electronics, electronic interfaces, and pres-surized gas controllers) or other devices. Such devicesmay be controlled using a control system 34.[0022] For systems including multiple spargers, controlsystem 34 may be operatively associated with each ofthe spargers and configured to operate the spargers in-dependently of each other. This can allow, for example,control of multiple gases being introduced into the con-tainer.[0023] The vessel may optionally include a mixing sys-tem such as an impeller 51, which can be rotated (e.g.,about a single axis) using a motor 52 that can be externalto the container. In some embodiments, as described inmore detail below, the impeller and motor are magneti-cally coupled. The mixing system can be controlled bycontrol system 34. Mixing systems are described in fur-ther detail below.[0024] Additionally and/or alternatively, the vessel mayinclude an antifoaming system such as a mechanical an-

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tifoaming device. As shown in the embodiment illustratedin Fig. 1, an antifoaming device may include, for example,an impeller 61 that can be rotated (e.g., magnetically)using a motor 62, which may be external to the container.The impeller can be used to collapse a foam containedin a head space 63 of the container. In some embodi-ments, the antifoaming system is in electrical communi-cation with a sensor 43 (e.g., a foam sensor) via controlsystem 34. The sensor may determine, for instance, thelevel or amount of foam in the head space or the pressurein the container, which can trigger regulation or controlof the antifoaming system. In other embodiments, theantifoaming system is operated independently of anysensors.[0025] The support structure and/or the container mayalso include, in some embodiments, one or more ports54 that can be used for sampling, analyzing (e.g., deter-mining pH and/or amount of dissolved gases in the liquid),or for other purposes. The support structure may alsoinclude one or more site windows 60 for viewing a levelof liquid within the container. One or more connections64 may be positioned at a top portion of the container orat any other suitable location. Connections 64 may in-clude openings, tubes, and/or valves for adding or with-drawing liquids, gases, and the like from the container,each of which may optionally include a flow sensor and/orfilter (not shown). The support structure may further in-clude a plurality of legs 66, optionally with wheels 68 forfacilitating transport of the vessel.[0026] It should be understood that not all of the fea-tures shown in Fig. 1 need be present in all embodimentsof the disclosure and that the illustrated elements maybe otherwise positioned or configured. Also, additionalelements may be present in other embodiments, suchas the elements described herein.[0027] Various aspects of the present invention are di-rected to a vessel including a container such as a col-lapsible bag. "Flexible container", "flexible bag", or "col-lapsible bag" as used herein, indicates that the containeror bag is unable to maintain its shape and/or structuralintegrity when subjected to the internal pressures (e.g.,due to the weight and/or hydrostatic pressure of liquidsand/or gases contained therein expected during opera-tion) without the benefit of a separate support structure.The collapsible bag may be made out of inherently flex-ible materials, such as many plastics, or may be madeout of what are normally considered rigid materials (e.g.,glass or certain metals) but having a thickness and/orphysical properties rendering the container as a wholeunable to maintain its shape and/or structural integritywhen subjected to the internal pressures expected duringoperation without the benefit of a separate support struc-ture. In some embodiments, collapsible bags include acombination of flexible and rigid materials; for example,the bag may include rigid components such as connec-tions, ports, supports for a mixing and/or antifoaming sys-tem, etc.[0028] The container (e.g., collapsible bag) may have

any suitable size for containing a liquid. For example, thecontainer may have a volume between 1-40L, 40-100 L,100-200 L, 200-300 L, 300-500 L, 500-750 L, 750-1,000L, 1,000-2,000 L, 2,000-5,000 L, or 5,000-10,000 L. Vol-umes greater than 10,000 L are also possible.[0029] In some embodiments, the collapsible bag isdisposable and is formed of a suitable flexible material.The flexible material may be one that is USP Class VIcertified, e.g., silicone, polycarbonate, polyethylene, andpolypropylene. Non-limiting examples of flexible materi-als include polymers such as polyethylene (e.g., linearlow density polyethylene and ultra low density polyethyl-ene), polypropylene, polyvinylchloride, polyvinyldichlo-ride, polyvinylidene chloride, ethylene vinyl acetate, poly-carbonate, polymethacrylate, polyvinyl alcohol, nylon,silicone rubber, other synthetic rubbers and/or plastics.As noted above, portions of the flexible container maycomprise a substantially rigid material such as a rigidpolymer (e.g., high density polyethylene), metal, and/orglass (e.g., in areas for supporting fittings, etc.). In otherembodiments, the container is substantially rigid materi-al. All or portions of the container may be optically trans-parent to allow viewing of contents inside the container.The materials or combination of materials used to formthe container may be chosen based on one or more prop-erties such as flexibility, puncture strength, tensilestrength, liquid and gas permeabilities, opacity, andadaptability to certain processes such as blow molding,injection molding, or spin cast molding (e.g., for formingseamless collapsible bags).[0030] The container (e.g., collapsible bag) may haveany suitable thickness for holding a liquid and may bedesigned to have a certain resistance to puncturing dur-ing operation or while being handled. For instance, thewalls of a container may have a total thickness of lessthan or equal to 250 mils (1 mil is 25.4 micrometers), lessthan or equal to 200 mils, less than or equal to 100 mils,less than or equal to 70 mils (1 mil is 25.4 micrometers),less than or equal to 50 mils, less than or equal to 25mils, less than or equal to 15 mils, or less than or equalto 10 mils. In some embodiments, the container includesmore than one layer of material that may be laminatedtogether or otherwise attached to one another to impartcertain properties to the container. For instance, one lay-er may be formed of a material that is substantially oxy-gen impermeable. Another layer may be formed of a ma-terial to impart strength to the container. Yet another layermay be included to impart chemical resistance to fluidthat may be contained in the container. It should be un-derstood that a container may be formed of any suitablecombinations of layers and that the invention is not limitedin this respect. The container (e.g., collapsible bag) mayinclude, for example, 1 layer, greater than or equal to 2layers, greater than or equal to 3 layers, or greater thanequal to 5 layers of material(s). Each layer may have athickness of, for example, less than or equal to 200 mils,less than or equal to 100 mils, less than or equal to 50mils, less than or equal to 25 mils, less than or equal to

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15 mils, less than or equal to 10 mils, less than or equalto 5 mils, or less than or equal to 3 mils, or combinationsthereof.[0031] In one set of embodiments of the disclosure,the container is seamless. The container may be, for ex-ample, a seamless collapsible bag or a seamless rigid(or semi-rigid) container. Many existing collapsible bagsare constructed from two sheets of a plastic materialjoined by thermal or chemical bonding to form a containerhaving two longitudinal seams. The open ends of thesheets are then sealed using known techniques and ac-cess apertures are formed through the container wall.During use, collapsible bags having seams can causethe formation of crevices at or near the seams wherefluids or reagents contained therein are not thoroughlymixed. In certain embodiments involving, for example,the use of collapsible bags for performing a chemical,biochemical and/or biological reaction, unmixed rea-gents can cause a reduction in yield of a desired product.The presence of the seams in a collapsible bag can alsoresult in the inability of the collapsible bag to conform tothe shape of a reusable support structure that may sup-port the bag. By using collapsible bags without any seamsjoining two or more flexible wall portions of the bag, how-ever, the problems of mixing and conformity may beavoided or reduced. In certain embodiments, seamlesscollapsible bags can be made specifically to fit a particularreusable support structure having a unique shape andconfiguration. Substantially perfect-fitting collapsiblebags can be used, for example, as part of a bioreactorsystem or a biochemical and/or chemical reaction sys-tem. Seamless rigid or semi-rigid containers may also bebeneficial in some instances.[0032] In certain embodiments, a collapsible bag thatdoes not include any seams joining two or more flexiblewall portions of the collapsible bag (i.e., a seamless col-lapsible bag) has a certain volume for containing a liquid.The seamless collapsible bag may have a volume of, forexample, at least 1 L, at least 10 liters, at least 20 liters,at least 40 liters, at least 50 liters, at least 70 liters, atleast 100 liters, at least 150 liters, at least 200 liters, atleast 300 liters, at least 500 liters, at least 700 liters, orat least 1,000 liters. Seamless collapsible bags may alsohave volumes greater than 1,000 liters (e.g., 1,000-5,000liters or 5,000-10,000 liters) as needed. In some embod-iments, the collapsible bag is positioned in a reusablesupport structure for surrounding and containing the flex-ible container.[0033] In one embodiment, a seamless collapsible bagis formed in a process in which the bag liner (e.g., theflexible wall portions of the bag), as well as one or morecomponents such as a component of an agitator/mixersystem (e.g., a shaft and/or a support base), port, etc. iscast from one continuous supply of a polymeric precursormaterial. In some cases, the casting may occur withouthermetically sealing, e.g., via welding. Such a seamlesscollapsible bag may allow the interior liquid or other prod-uct to contact a generally even surface, e.g., one which

does not contain substantial wrinkles, folds, crevices, orthe like. In addition, in some cases, the collapsible bagcomplementarily fits within a support structure when in-stalled and filled with a liquid or product. The seamlesscollapsible bag may also have a generally uniform poly-meric surface chemistry which may, for example, mini-mize side reactions. Methods of forming seamless col-lapsible bags involving more than one polymeric precur-sor materials can also be performed.[0034] Seamless collapsible bags can be created by avariety of methods. In one embodiment, a seamless col-lapsible bag is formed by injecting liquid plastic into amold that has been pre-fitted with components such asports, connections, supports, and rigid portions config-ured to support a mixing system (e.g., a shaft and/or abase) that are subsequently surrounded, submerged,and/or embedded by the liquid plastic. The componentmay be a rigid component, e.g., one that can substantiallymaintain its shape and/or structural integrity during use.Any suitable number of components (e.g., at least 1, 2,5, 10, 15, etc.) can be integrated with containers (e.g.,collapsible bags) using methods described herein. Themold may be designed to form a collapsible bag havingthe shape and volume of the mold, which may have asubstantially similar shape, volume, and/or configurationof a reusable support structure.[0035] In one embodiment, the container is formed byusing an embedded component/linear molding (ECM)technique. In one such technique, rigid or pre-made com-ponents such as tube ports, agitator bases, etc. are firstpositioned in the mold. A polymer or polymer precursorused to form a container (e.g., a seamless collapsiblebag) may be introduced (e.g., in a melt state) via a pol-ymer fabrication technique such as those described be-low. In some cases, a component or a portion of the com-ponent is partially melted by the polymer precursor, al-lowing the component to form a continuous element withthe container. That is, the component can be joined (e.g.,fused) with one or more wall portions of the container(e.g., flexible wall portions of a collapsible bag) to forma single, integral piece of material without seams.[0036] Referring now to Fig. 2A, an illustrative exampleof this process is shown. In this figure, a premountedcomponent 202 is held between mold walls 205 and 206.Polymer precursor 210 is then introduced into the moldvia any suitable technique. The polymer precursor thenflows around component 202 and is hardened, solidified,set, or cured, thereby forming a seamless connectionbetween the polymer itself and the embedded compo-nent.[0037] In some cases, components are designed withthinner portions that can be melted with a polymer pre-cursor (e.g., in the melt state) during formation of a con-tainer. For example, as shown in the embodiment illus-trated in Fig. 2B, component 203, a rigid portion includinga recess 212 into which at least a portion of a drive head(not shown) of a mixing and/or antifoaming system canbe inserted, is positioned in a mold comprising a shape

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configured to mold a container. Polymer precursor 210is then introduced into the mold via any suitable tech-nique. The polymer precursor then flows around compo-nent 203 and melts thinner portions 204, causing fusionof portions of the walls of the container and portions ofthe component. The container and component are thenhardened, solidified, set, or cured, thereby forming a sin-gle, integral piece of material where a seamless connec-tion exists between the polymer itself and the embeddedcomponent. This technique can be used to form, for ex-ample, a container (e.g., a seamless collapsible bag)having a volume of greater than, e.g., 40 L, 50 L, 100 L,200 L, 1,000 L, etc., and having embedded therein com-ponents such as one or more support bases and/or shaftsfor impellers of a mixing system(s).[0038] Accordingly, one embodiment of the disclosureincludes the method of joining together a wall portion ofa container and at least a portion of a functional compo-nent during formation of the container within a mold,wherein the portion of the functional component is meltedduring the joining step. The wall portion of the containermay have a first thickness and the portion of the functionalcomponent may have a second thickness, the thickness-es being within, for example, less than 100%, 80%, 60%,40%, 20%, 10%, or 5% of each other, relative to the largerof the first and second thicknesses.[0039] In another embodiment, the container may beformed using a continuous component/liner molding(CCM) technique. In one such technique, a collapsiblebag or other container is cast de novo from a polymer orpolymer precursor stream. The polymer or polymer pre-cursor used to form the seamless collapsible bag is in-troduced via a polymer fabrication technique such asthose described below. Components can be introducedinto the flexible container by using mandrels, barriers,baffles, and the like to direct the polymer precursor toform functional components of a liquid containment sys-tem such as tube ports and agitator bases as, for exam-ple, one continuous polymer. After setting or curing ofthe polymer or polymer precursor, the mandrels, barriers,etc., may be withdrawn. For example, as is shown in Fig.2C, mandrel 209 is held between mold walls 205 and206. Polymer 210 is then introduced into the mold viaany suitable technique, such as those described below.The polymer then flows around mandrel 209 and is hard-ened or set around the mandrel. The mandrel can thenbe subsequently withdrawn. This technique can be usedto form, for example, a container (e.g., a seamless col-lapsible bag) having a volume of greater than, e.g., 40L, 50 L, 100 L, 200 L, 1,000 L, etc., having combinedtherein components (e.g., rigid components) that arejoined to the collapsible bag without welding.[0040] Combinations of these and/or other techniquesmay also be used in other embodiments. For instance,in some cases, different polymer formulations (such aslow molecular weight polyethylene, high molecularweight polyethylene, polypropylene, silicone, polycar-bonate, polymethacrylate, combinations thereof or pre-

cursors thereof) can be simultaneously injected into re-gions of the mold designed to form a more rigid structuresuch as tubing or sensor ports, agitation systems, etc. Inone particular embodiment, a method involves introduc-ing a first polymer or polymer precursor into a mold com-prising a shape configured to mold a collapsible bag hav-ing a volume of at least 10 mL, 1L, 40 L, 100 L, or 1,000L, etc. The mold may further comprise a shape configuredto mold a component of a mixing and/or antifoaming sys-tem such as a shaft and/or base configured to supportan impeller. The method may also include introducing asecond polymer or polymer precursor into the mold tomold the component of the mixing system. Accordingly,the component of the mixing system and the collapsiblebag may be joined without welding using methods de-scribed herein. In some instances, the first and secondpolymers or polymer precursors are introduced simulta-neously. The first and second polymers or polymer pre-cursors may be the same in some embodiments, or dif-ferent in other embodiments. Such a method can be usedto form, for example, base plates for mixing/agitation sys-tems, antifoaming systems, or other components. In oth-er embodiments, a number of polymers can be intro-duced into the mold (e.g., simultaneously) to form con-tainers with multiple components.[0041] As mentioned, a polymer or polymer precursormay be introduced into a mold to form a container suchas a collapsible bag (e.g., a seamless collapsible bag)using any suitable technique. For instance, in one em-bodiment, the collapsible bag may be fabricated via aspin casting process. For example, during spin casting,a mold may be spun during injection of the polymer orpolymeric precursor to deposit a uniform coating of plas-tic on the mold surface. In another embodiment, the col-lapsible bag is fabricated via an injection molding proc-ess. For instance, the polymeric precursor may bepumped into the space between an inner mold and theouter mold. In yet another embodiment, the collapsiblebag can be fabricated via a blow molding process. Thepolymer may be deposited, for example, via a gas injec-tion, to expand the polymer against the mold surface. Inyet another embodiment, a combination of these and/orother techniques may be used. Those of ordinary skill inthe art will be familiar with polymer processing techniquessuch as spin casting, injection molding, and/or blow mold-ing, and will be able to use such techniques in the meth-ods of the present invention as described herein to pre-pare suitable collapsible bags or other containers.[0042] Although many embodiments herein describeseamless collapsible bags, in some embodiments, col-lapsible bags or other containers described herein canbe fabricated with seams between flexible wall portionsof the container. In other embodiments, collapsible bagsor other containers can be fabricated with seams be-tween a component and one or more flexible wall portionsof the container. The act of joining two or more wall por-tions or a wall portion and a portion of a component maybe achieved by methods such as welding (e.g., heat weld-

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ing and ultrasonic welding), bolting, use of adhesives,fastening, or other attaching techniques. Combination ofseams and seamless connections can also be fabricated.It should also be understood that while many of the meth-ods described herein refer to fabrication of collapsiblebags, the methods may also be applied to rigid contain-ers. The methods described herein used to form contain-ers such as collapsible bags (e.g., bags with or withoutseams) may be adapted to include components of vari-ous sizes. For instance, although the flexible wall portionsof a collapsible bag may having a thickness of, for exam-ple, less than or equal to 100 mils, less than or equal to70 mils, less than or equal to 50 mils, less than or equalto 25 mils, less than or equal to 15 mils, or less than orequal to 10 mils, a component to be incorporated withthe container may have a thickness or a height of, forexample, greater than 0.5 mm, greater than 1 cm, greaterthan 1.5 cm, greater than 2 cm, greater than 5 cm, orgreater than 10 cm. In some cases, the component hasat least one cross-sectional dimension (e.g., a height,length, width, or diameter) of, for example, greater than0.5 mm, greater than 1 cm, greater than 1.5 cm, greaterthan 2 cm, greater than 5 cm, greater than 10 cm, greaterthan 15 cm, greater than 20 cm, greater than 25 cm, orgreater than 30 cm. In certain embodiments, the thick-ness of a collapsible bag (or other container) and thethickness of a portion of a component to be joined (e.g.,fused) with the collapsible bag are within 30%, 20%, 15%,10% or 5% of each other (relative to the thickest portion).This matching of thicknesses can aid joining (e.g., melt-ing, welding, etc.) of the materials, as described in moredetail below.[0043] Components that are integrated with collapsiblebags or other containers may be formed in any suitablematerial, which may be the same or a different materialfrom that of the bag or container. For instance, in oneembodiment, a container is formed in a first polymer anda component is formed in a second polymer that is dif-ferent (e.g., having a different composition, molecularweight, and/or chemical structure, etc.) from the first pol-ymer. Those of ordinary skill in the art will be familiar withmaterial processing techniques and will be able to usesuch techniques in the methods described herein tochoose suitable materials and combinations of materials.[0044] In some embodiments, components that are in-tegrated with collapsible bags or other containers usingmethods described herein are formed in one or morematerials that is/are USP Class VI certified, e.g., silicone,polycarbonate, polyethylene, and polypropylene. Non-limiting examples of materials that can be used to formcomponents include polymers such as polyethylene(e.g., low density polyethylene and high density polyeth-ylene), polypropylene, polyvinylchloride, polyvinyldichlo-ride, polyvinylidene chloride, ethylene vinyl acetate, pol-yvinyl alcohol, polycarbonate, polymethacrylate, nylon,silicone rubber, other synthetic rubbers and/or plastics,and combinations thereof. Ceramics, metals, and mag-netic materials can also be used to form all or portions

of a component. In some embodiments, all or portions ofa component are rigid; in other embodiments, all or por-tions of a component are flexible. The material(s) usedto form a component may be chosen based on, for ex-ample, the function of the component and/or one or moreproperties such as compatibility with the container, flex-ibility, tensile strength, hardness, liquid and gas perme-abilities, and adaptability to certain processes such asblow molding, injection molding, or spin cast molding.[0045] In certain embodiments, especially in certainembodiments involving fluid manipulations or carryingout a chemical, biochemical and/or biological reaction ina container (e.g., a collapsible bag), the container is sub-stantially closed, e.g., the container is substantiallysealed from the environment outside of the container ex-cept, in certain embodiments, for one or more inlet and/oroutlet ports that allow addition to, and/or withdrawal ofcontents from, the container. If a collapsible bag is used,it may be substantially deflated prior to being filled witha liquid, and may begin to inflate as it is filled with liquid.In other embodiments, aspects of the invention can beapplied to opened container systems.[0046] In some cases, fluids may be introduced and/orremoved from a vessel via inlet ports and/or outlet ports.The vessel may be a part of a reactor system for per-forming a biological, biochemical, or chemical reaction.As mentioned, a container (e.g., a collapsible bag), whichmay be part of the vessel, may have any suitable numberof inlet ports and any suitable number of outlet ports. Insome cases, pumps, such as disposable pumps, may beused to introduce a gas or other fluid into the container,e.g., via an inlet port, and/or which may be used to removea gas or other fluid from the container, e.g., via an outletport. For instance, a magnetically-coupled pump may becreated by encasing a disposable magnetic impellerhead in an enclosure with inlet(s) and outlet(s) thatachieves fluid pumping. Flexible blades may be used toenhance pumping or provide pressure relief. In anotherembodiment, pumping of fluids, gas and/or powder maybe achieved without pump heads and/or pump chambersby sequentially squeezing, for example, an electrome-chanical-polymeric tube that effectively achieves "peri-stalsis." One way valves in the tube may optionally beused, which may aid in the prevention of backflow.[0047] Certain aspects of the present invention alsoinclude a support structure, for example, support struc-ture 14 as shown in Fig. 1, which can surround and con-tain container 18. The support structure may have anysuitable shape able to surround and/or contain the con-tainer. In some cases, the support structure is reusable.The support structure may be formed of a substantiallyrigid material. Non-limiting examples of materials that canbe used to form the reusable support structure includestainless steel, aluminum, glass, resin-impregnated fib-erglass or carbon fiber, polymers (e.g., high-density pol-yethylene, polyacrylate, polycarbonate, polystyrene, ny-lon or other polyamides, polyesters, phenolic polymers,and combinations thereof. The materials may be certified

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for use in the environment in which it is used. For exam-ple, non-shedding materials may be used in environ-ments where minimal particulate generation is required.[0048] In some embodiments, the reusable supportstructure may be designed to have a height and diametersimilar to standard stainless steel bioreactors (or otherstandard reactors or vessels). The design may also bescaleable down to small volume bench reactor systems.Accordingly, the reusable support structure may haveany suitable volume for carrying out a desired chemical,biochemical and/or biological reaction. In many instanc-es, the reusable support structure has a volume substan-tially similar to that of the container. For instance, a singlereusable support structure may be used to support andcontain and single container having a substantially sim-ilar volume. In other cases, however, a reusable supportstructure is used to contain more than one container. Thereusable support structure may have a volume between,for example, 1-100 L, 100-200 L, 200-300 L, 300-500 L,500-750 L, 750-1,000 L, 1,000-2,000 L, 2,000-5,000 L,or 5,000-10,000 L. Volumes greater than 10,000 L arealso possible.[0049] In other embodiments, however a vessel of thepresent invention does not include a separate container(e.g., collapsible bag) and support structure, but insteadcomprises a self-supporting disposable container. Thecontainer may be, for example, a plastic vessel and, insome cases, may include an agitation system integrallyor removably attached thereto. The agitation system maybe disposable along with the container. In one particularembodiment, such a system includes an impeller weldedor bolted to a polymeric container. It should therefore beunderstood that many of the aspects and features of thevessels described herein with reference to a containerand a support structure (for example, a seamless con-tainer, a sparging system, an antifoaming device, etc.),are also applicable to a self-supporting disposable con-tainer.[0050] As an example, a container, such as container18 shown in Fig. 3, may include various sensors and/orprobes for controlling and/or monitoring one or moreprocess parameters inside the disposable container suchas, for example, temperature, pressure, pH, dissolvedoxygen (DO), dissolved carbon dioxide (DCO2), mixingrate, and gas flow rate. The sensor may also be an opticalsensor in some cases.[0051] In some embodiments, process control may beachieved in ways which do not compromise the sterilebarrier established by a disposable container. For exam-ple, gas flow may be monitored and/or controlled by arotameter or a mass flow meter upstream of an inlet airfilter. In another embodiment, disposable optical probesmay be designed to use "patches" of material containingan indicator dye which can be mounted on the inner sur-face of the disposable container and read through thewall of the disposable container via a window in the re-usable support structure. For example, dissolved oxy-gen, pH, and/or CO2 each may be monitored and con-

trolled by an optical patch and sensor mounted on, e.g.,a gamma-irradiatable, biocompatible polymer which, canbe sealed to, embedded in, or otherwise attached to thesurface of the container.[0052] As a specific example of a sensor, as shown inthe embodiment illustrated in Fig. 3, container 18 maybe operatively associated with a temperature controller106 which may be, for example, a heat exchanger, aclosed loop water jacket, an electric heating blanket, ora Peltier heater. Other heaters for heating a liquid insidea container are known to those of ordinary skill in the artand can also be used in combination with container 18.The heater may also include a thermocouple and/or aresistance temperature detector (RTD) for sensing a tem-perature of the contents inside the container. The ther-mocouple may be operatively connected to the temper-ature controller to control temperature of the contents inthe container. Optionally, a heat-conducting materialmay be embedded in the surface of the container to pro-vide a heat transfer surface to overcome the insulatingeffect of the material used to form other portions of thecontainer.[0053] In general, as used herein, a component of aninventive system that is "operatively associated with" oneor more other components indicates that such compo-nents are directly connected to each other, in direct phys-ical contact with each other without being connected orattached to each other, or are not directly connected toeach other or in contact with each other, but are mechan-ically, electrically (including via electromagnetic signalstransmitted through space), or fluidically interconnectedso as to cause or enable the components so associatedto perform their intended functionality.[0054] Cooling may also be provided by a closed loopwater jacket cooling system, a cooling system mountedon the reactor, or by standard heat exchange through acover/jacket on the reusable support structure (e.g., theheat blanket or a packaged dual unit which provides heat-ing and cooling may a component of a device configuredfor both heating/cooling but may also be separate froma cooling jacket). Cooling may also be provided by Peltiercoolers. For example, a Peltier cooler may be applied toan exhaust line to condense gas in the exhaust air to helpprevent an exhaust filter from wetting out.[0055] In certain embodiments, a reactor system in-cludes gas cooling for cooling the head space and/or exitexhaust. For example, jacket cooling, electrothermaland/or chemical cooling, or a heat exchanger may beprovided at an exit air line and/or in the head space ofthe container. This cooling can enhance condensate re-turn to the container, which can reduce exit air filter plug-ging and fouling. In some embodiments, purging of pre-cooled gas into the head space can lower the dew pointand/or reduce water vapor burden of the exit air gas.[0056] In some cases, sensors and/or probes (e.g.,probe 106) may be connected to a sensor electronicsmodule 132, the output of which can be sent to a terminalboard 130 and/or a relay box 128. The results of the sens-

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ing operations may be input into a computer-implement-ed control system 115 (e.g., a computer) for calculationand control of various parameters (e.g., temperature andweight/volume measurements) and for display and userinterface. Such a control system may also include a com-bination of electronic, mechanical, and/or pneumatic sys-tems to control heat, air, and/or liquid delivered to or with-drawn from the disposable container as required to sta-bilize or control the environmental parameters of theprocess operation. It should be appreciated that the con-trol system may perform other functions and the inventionis not limited to having any particular function or set offunctions.[0057] The one or more control systems can be imple-mented in numerous ways, such as with dedicated hard-ware and/or firmware, using a processor that is pro-grammed using microcode or software to perform thefunctions recited above or any suitable combination ofthe foregoing. A control system may control one or moreoperations of a single reactor for a biological, biochemicalor chemical reaction, or of multiple (separate or intercon-nected) reactors.[0058] Each of systems described herein (e.g., withreference to Fig. 3), and components thereof, may beimplemented using any of a variety of technologies, in-cluding software (e.g., C, C#, C++, Java, or a combinationthereof), hardware (e.g., one or more application-specificintegrated circuits), firmware (e.g., electrically-pro-grammed memory) or any combination thereof.[0059] Various embodiments according to the disclo-sure may be implemented on one or more computer sys-tems. These computer systems, may be, for example,general-purpose computers such as those based on IntelPENTIUM-type and XScale-type processors, MotorolaPowerPC, Motorola DragonBall, IBM HPC, Sun Ul-traSPARC, Hewlett-Packard PA-RISC processors, anyof a variety of processors available from Advanced MicroDevices (AMD) or any other type of processor. It shouldbe appreciated that one or more of any type of computersystem may be used to implement various embodimentsof the invention. The computer system may include spe-cially-programmed, special-purpose hardware, for ex-ample, an application-specific integrated circuit (ASIC).Aspects of the invention may be implemented in soft-ware, hardware or firmware, or any combination thereof.Further, such methods, acts, systems, system elementsand components thereof may be implemented as part ofthe computer system described above or as an independ-ent component.[0060] In one embodiment, a control system operative-ly associated with a vessel described herein is portable.The control system may include, for example, all or manyof the necessary controls and functions required to per-form a fluidic manipulation (e.g., mixing and reactions)in the control system. The control system may include asupport and wheels for facilitating transport of the vessel.Advantageously, such a portable control system can beprogrammed with set instructions, if desired, transported

(optionally with the vessel), and hooked up to a vessel,ready to perform a fluid manipulation in a shorter amountof time than conventional fluid manipulation control sys-tems (e.g., less than 1 week, 3 days, 1 day, 12 hours, 6hours, 3 hours, or even less than 1 hour).[0061] A vessel, including the container, may also beconnected to one or more sources of gases such as air,oxygen, carbon dioxide, nitrogen, ammonia, or mixturesthereof, in various aspects of the invention. The gasesmay be compressed, pumped, etc. Such gases may beused to provide suitable growth and/or reaction condi-tions for producing a product inside the container. Thegases may also be used to provide sparging to the con-tents inside the container, e.g., for mixing or other pur-poses. For instance, in certain embodiments employingspargers, bubble size and distribution can be controlledby passing an inlet gas stream through a porous surfaceprior to being added to the container. Additionally, thesparging surface may be used as a cell separation deviceby alternating pressurization and depressurization (orapplication of vacuum) on the exterior surface of the po-rous surface, or by any other suitable method.[0062] As a specific example, Fig. 3 shows varioussources of gases 118 and 124. The inlet gases may op-tionally pass through filter 120 and/or a flow meter and/orvalve 122, which may be controlled by controller system115, prior to entering the container. Valve 122 may be apneumatic actuator (actuated by, e.g., compressedair/carbon dioxide or other gas 124), which may be con-trolled by a solenoid valve 126. These solenoid valvesmay be controlled by a relay 128 connected to terminalboard 130, which is connected to the controller system115. The terminal board may comprise, for example, aPCI terminal board, or a USB/parallel, or fire port terminalboard of connection. In other embodiments, flush closingvalves can be used for addition ports, harvest and sam-pling valves. Progressive tubing pinch valves that areable to meter flow accurately can also be used. In somecases, the valves may be flush closing valves (e.g., forinlet ports, outlet ports, sampling ports, etc.). The inletgases may be connected to any suitable inlet of the ves-sel. In one embodiment, the inlet gases are associatedwith one or more spargers which can be controlled inde-pendently, as described in more detail below.[0063] As shown in the exemplary embodiment illus-trated in Fig. 3, the container and support structure illus-trated in Fig. 1 can be operatively associated with a va-riety of components as part of an overall bioreactor sys-tem 100, according to another aspect of the invention.Accordingly, the container and/or support structure mayinclude several fittings to facilitate connection to function-al component such as filters, sensors, and mixers, aswell as connections to lines for providing reagents suchas liquid media, gases, and the like. The container andthe fittings may be sterilized prior to use so as to providea "sterile envelope" protecting the contents inside thecontainer from airborne contaminants outside. In someembodiments, the contents inside the container do not

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contact the reusable support structure and, therefore, thereusable support structure can be reused after carryingout a particular chemical, biochemical and/or biologicalreaction without being sterilized, while the containerand/or fittings connected to the container can be discard-ed. In other embodiments, the container, fittings, and/orreusable support structure may be reused (e.g., aftercleaning and sterilization).[0064] A vessel may also include a mixing system formixing contents of the container, in another aspect. Insome cases, more than one agitator or mixer may beused, and the agitators and/or mixes may the same ordifferent. More than one agitation system may be used,for example, to increase mixing power. In some cases,the agitator may be one in which the height can be ad-justed, e.g., such that the draft shaft allows raising of animpeller or agitator above the bottom of the tank and/orallows for multiple impellers or agitators to be used. Amixing system of a vessel may be disposable or intendedfor a single use (e.g., along with the container), in somecases.[0065] Various methods for mixing fluids can be imple-mented in the container. For instance, mixers based onmagnetic actuation, sparging, and/or air-lift can be used.Direct shaft-drive mixers that are sealed and not mag-netically coupled can also be used. In one particular em-bodiment, mixing systems such as the ones disclosed inU.S. Patent Application Serial No. 11/147, 124, filed June6, 2005, entitled "Disposable Bioreactor Systems andMethods," by G. Hodge, et al., published as U.S. PatentApplication Publication No. 2005/0272146 on December8, 2005, are used with embodiments described herein.For example, the mixing system may include a motor112, e.g., for driving an impeller (or other componentused for mixing) positioned inside the container, a powerconditioner 114, and/or a motor controller 116.[0066] In some cases, a plurality (e.g., more than 1, 2,or 3, etc.) of mixers or impellers are used for mixing con-tents in a container. Additionally and/or alternatively, amixing system may include an adjustable height impellerand/or an impeller with varying impeller blade configura-tions. For instance, the mixer may have an extended driveshaft which allows the impeller to be raised to differentheights relative to the bottom of the container. The ex-tended shaft can also allow integration of multiple impel-lers. In another embodiment, a bioreactor system in-cludes more than one agitation drive per container, whichcan increase mixing power.[0067] To enhance mixing efficiency, the containermay include baffles such as internal film webs or protru-sions, e.g., positioned across the inside of the containeror extending from the inner surface of the container atdifferent heights and at various angles. The baffles maybe formed of in any suitable material such as a polymer,a metal, or a ceramic so long as they can be integratedwith the container.[0068] In one embodiment, a direct drive agitator isused. Typically, the agitator includes a direct shaft drive

that is inserted into the container. In certain instances,the location where the shaft exits the container may bemaintained in a sterile condition. For instance, internaland/or external rotating seals may be used to maintaina sterile seal, and/or live hot steam may be used to fa-cilitate maintenance of the sterile seal. By maintainingsuch a sterile seal, contamination caused by the shaft,e.g., from the external environment, from the exiting gas-es, etc., may be reduced or avoided.[0069] In another embodiment, a magnetic agitator isused. Typically, a magnetic agitator uses magnets suchas fixed or permanent magnets to rotate or otherwisemove the agitator, for example, impellers, blades, vanes,plates, cones, etc. In some cases, the magnets withinthe magnetic agitator are stationary and can be turnedon or activated in sequence to accelerate or deceleratethe agitator, e.g., via an inner magnetic impeller hub. Asthere is no penetration of the container by a shaft, theremay be no need to maintain the agitator in a sterile con-dition, e.g., using internal and/or external rotating seals,live hot steam, or the like.[0070] In yet another embodiment, an electromechan-ical polymeric agitator is used, e.g., an agitator that in-cludes an electromechanical polymer-based impellerthat spins itself by "paddling," i.e., where the agitator ismechanically flapped to propel the agitator or impeller,e.g., rotationally.[0071] Specific non-limiting examples of devices thatcan be used as a mixing system, and/or an antifoamingsystem in certain embodiments, are illustrated in Figs.4A-8. The devices shown include a magnetically-actuat-ed impeller, although other arrangements are possible.In some of these magnetic configurations, the motor isnot directly connected to the impeller. Magnets associ-ated with a drive head can be aligned with magnets as-sociated with an impeller hub, such that the drive headcan rotate the impeller through magnetic interactions. Insome cases, the motor portion (and other motor associ-ated components) may be mounted on the support struc-ture.[0072] As shown in Fig. 4A, this exemplary systemgenerally includes an impeller support 300 affixed to por-tions of a container wall 302, preferably at a lower portionthereof, an impeller hub 304, a motor 306, a motor shaft308 and a drive head 310. The impeller support may beaffixed to the wall of the container using any suitable tech-nique, e.g., by heat welding together two portions of atwo-piece impeller support, sandwiching the containerwall therebetween or onto the wall, or using other meth-ods described herein. As one example, an opening in thewall of the container may be used to allow a central por-tion of the impeller plate to extend from an exterior of thecontainer to the interior (or vise versa). Then a sealingring (not shown) may be adhered or the container maybe welded directly to an outer circumference of the im-peller support to seal the container wall therebetween.As another example, an undersized opening in the wallof the container may be used to form a seal with a cir-

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cumferencial edge of the impeller support slightly largerthan the opening. In other embodiments, at least a portionof the impeller support is embedded with a wall of thecontainer and/or the impeller support and container arefabricated simultaneously (e.g., by spin casting, injectionmolding, or blow molding).[0073] One feature according to one embodiment ofthe disclosure is directed to the inclusion of one or morespargers associated with an impeller support, which maybe used to direct air or other gases into the container. Insome cases, the sparger may include porous, micro-po-rous, or ultrafiltration elements 301 (e.g., sparging ele-ments). The spargers may be used to allow a gaseoussparge or fluids into and/or out of the container by beingdimensioned for connection to a source of a gas; thisconnection may take place via tubing 306. Such spargingand/or fluid addition or removal may be used, in somecases, in conjunction with a mixing system (e.g., the ro-tation of the impeller hub). Sparging systems are de-scribed in more detail below.[0074] In the embodiment illustrated in Fig. 4A, the in-terior side of the impeller support may include a shaft orpost 312 to which a central opening in the impeller hub304 receives. The impeller hub may be maintained at aslight distance 305 above the surface of the impeller sup-port (e.g., using a physical spacer) to prevent friction ther-ebetween. Low friction materials may be used in the man-ufacture of the impeller hub to minimize friction betweenthe impeller hub and the post. In another embodiment,one or more bearings may be included to reduce friction.For instance, the impeller hub may include, in certaininstances, a bearing 323 (e.g., a roller bearing, ball bear-ing (e.g., a radial axis ball bearing), thrust bearing, racebearing, double raceway bearing, lazy-susan bearing, orany other suitable bearing) for reducing or preventingfriction between the impeller support and the post. Addi-tionally, the drive head may include a physical spacer324 for reducing or preventing friction between the drivehead and the impeller support.[0075] The impeller hub also may include one or moremagnets 314, which may be positioned at a periphery ofthe hub or any other suitable position, and may corre-spond to a position of a magnet(s) 316 provided on thedrive head 310. The poles of the magnets may be alignedin a manner that increases the amount of magnetic at-traction between the magnets of the impeller hub andthose of the drive head.[0076] The drive head 310 may be centrally mountedon a shaft 308 of motor 306. The impeller hub also mayinclude one or more impeller blades 318. In some cases,the embedded magnet(s) in the impeller can also be usedto remove ferrous or magnetic particles from solutions,slurries, or powders.[0077] An example of such a system is described inmore detail in U.S. Patent Application Serial No.11/147,124, filed June 6, 2005, entitled "Disposable Bi-oreactor Systems and Methods," by G. Hodge, et al.,published as U.S. Patent Application Publication No.

2005/0272146 on December 8, 2005.[0078] Fig. 4B illustrates another embodiment, havinga mechanically-driven impeller. As shown, this embodi-ment generally includes an impeller support 400, an im-peller hub 404 with shaft 405, and an external motor 406with shaft 408. The connection of shafts between theimpeller hub shaft and the motor shaft may be accom-plished in a matter familiar to one of ordinary skill in theart (e.g., gear box, hex drive, or the like).[0079] The impeller support can be affixed, for in-stance, to a side of the bioreactor wall 402 at a lowerportion thereof. The impeller support may be affixed tothe wall of the bioreactor by any of the methods discussedherein. Porous, micro-porous, or ultrafiltration elements401 may also be included in the present embodiment toallow gaseous sparge or fluids into and out of the biore-actor, as discussed in detail below. In the embodimentillustrated in Fig. 4B, the shaft of the impeller hub maybe received in a seal 412 (which may also include a bear-ing, in some cases) centrally located in an impeller sup-port 400. The seal can be used to insure that the contentsof the container are not contaminated. The impeller hubcan also be maintained at a slight distance above thesurface of the impeller support to prevent friction there-between. The impeller hub may include one or more im-peller blades 418, or other suitable mixing structures,such as vanes, plates, cones, etc.[0080] One aspect of the disclosure involves the rec-ognition that a careful and close alignment, vertically andhorizontally, between the drive head and impeller supportcan add significant benefits to devices of the invention.Prior to the invention, this may not have been recognizedor appreciated as indicated by the general acceptanceof the utility, and potential lack of need for improvement,of typical magnetic stirring/rotating mechanisms for flu-ids. An example includes traditional magnetic stir bar ar-rangements in chemical and biochemical laboratories,where it is typically quite acceptable for magnetic stirrermotors, and stir bars located within a reaction flask, tobe not carefully aligned or carefully positioned in proxim-ity to each other. The present invention involves recog-nition that better positioning and proximity can beachieved through techniques of the invention and resultin better potential stirring torque and/or stirring rotationalrates.[0081] Referring now to Fig. 5, one embodiment of adrive head magnetically coupled to an impeller of theinvention is illustrated schematically. In Fig. 5, an impellersupport 501, shown in a cross-section, includes a sub-stantially horizontal portion 504, from which a substan-tially vertical impeller shaft 508 extends upwardly sup-porting an impeller 509 (which may include a core 510and blades 511). Impeller 509 may rotate about shaft508. Optionally, this rotation may be facilitated by a bear-ing 507, which may be any suitable bearing such as aroller bearing, ball bearing (e.g., a radial axis ball bear-ing), thrust bearing, race bearing, double raceway bear-ing, lazy-susan bearing, or the like. Impeller support 501

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includes drive head alignment elements 512 which, inthe embodiment illustrated, are substantially verticaldownwardly-depending ridges which can define a circu-lar recess into which at least a portion of a drive head516 can be inserted. Guide elements 512 are positionedsuch that drive head, when engaged with the impellersupport, position the drive head at a predetermined de-sired location relative impeller 509. In one arrangement,guide elements 512 center the drive head, when engagedwith the impeller support, with respect to impeller 509.As a further, optional embodiment, a physical spacer 520can be provided between drive head 516 and a bottomsurface 524 of the impeller support aligned with that por-tion of the top surface 526 of the drive head at the locationat which the drive head is ideally positioned with respectto the impeller support. Physical spacer 520 physicallyseparates, by a desired distance, the bottom surface 524of the impeller support with a top surface 526 of the drivehead, but, at least one portion between the top surfaceof the drive head and bottom surface of the impeller sup-port, may define a continuous, physical connection (freeof voids of air or the like), between the drive head andthe impeller support. This allows for closer tolerance ofthe drive head with the impeller support than would havebeen realized in many prior arrangements, and it allowsfor reproducible and secure engagement of the drivehead with the impeller support. In some cases, the drivehead includes a recess 528 into which at least a portionof physical spacer 520 can be inserted. This arrangementcan allow reproducible and secure engagement of thedrive head with the physical spacer.[0082] The bottom of the impeller support and the topsurface of the drive head can be separated (e.g., usinga physical spacer) by a distance 521. In one embodiment,distance 521 is no greater than 50% of average thickness530 of the substantially horizontal portion 504 of the im-peller support. In other embodiments, this distance is nomore than 40%, 30%, 20%, 10%, or 5% of the thicknessof the impeller support.[0083] In some embodiments, physical spacer 520 hasa thickness no greater than 50% of average thickness530 of the substantially horizontal portion 504 of the im-peller support. In other embodiments, this thickness isno more than 40%, 30%, 20%, 10%, or 5% of the thick-ness of the impeller support.[0084] In one set of embodiments, physical spacer 520is a bearing that facilitates rotation of the drive head rel-ative to the impeller support. Where physical spacer 520is a bearing, any suitable bearing can be selected suchas a roller bearing, ball bearing (e.g., a radial axis ballbearing), thrust bearing, race bearing, double racewaybearing, lazy-susan bearing, or the like.[0085] In the embodiment illustrated in Fig. 5, the drivehead can vary in position, relative to shaft 508, horizon-tally no more than 5 mm during normal operation or, inother embodiments, no more than 4, 3, 2, 1 (0.5, or 0.25mm during normal operation). The drive head can alsovary in distance relative to bottom surface 524 of the im-

peller support by no more than 10 mm, 1 mm, 0.5 mm,0.25 mm, 0.1 mm, or 0.005 mm in certain embodimentswith the use of the arrangements illustrated in Fig. 5.[0086] The arrangements of Fig. 5, especially in em-bodiments where physical spacer 520 is used, also addsphysical support to impeller support 501 in addition toany other physical support which the impeller support501 might receive. This added support is particularly ad-vantageous in collapsible bag arrangements includingimpellers (e.g., for mixers and/or antifoaming devices).[0087] Optionally, impeller support 501 may includespargers 540 positioned beneath blades of the impeller.The spargers can be dimensioned for connection to oneor more sources of gas. For example, the spargers mayinclude a port that can be connected to tubing 542 in fluidcommunication with one or more sources of gas.[0088] Although many of the figures described hereinshow impellers that are positioned at or near a bottomportion of a container, in other embodiments, impellerscan be positioned at any suitable location within a con-tainer, for example, near the center or a top portion of acontainer. This can be achieved by extending the lengthof a shaft which supports the impeller, or by any othersuitable configuration. Positions of impellers in a contain-er may depend on the process to be performed in thecontainer. For instance, in some embodiments wheresparging is required, impellers may be positioned nearthe sparger such that the impeller can sweep and/or reg-ulate the bubbles introduced into the container. Addition-ally, although the figures described herein show a singleimpeller associated with a shaft, more than one impellercan be used in some instances. For example, a first im-peller coupled to a shaft may be located near a bottomportion of the container and a second impeller coupledto the shaft may be positioned near the center of thecontainer. The first impeller may provide adequatesweeping of a sparged gas, and the second impeller mayprovide adequate mixing of contents within the container.[0089] In one aspect of the disclosure, the impeller sup-port is uniquely designed to be readily fastenable to acollapsible bag. Certain know arrangements of impellersattached to collapsible bags may suffer from drawbacksresulting from non-ideal attachment of the bag to the im-peller support, or non-ideal techniques for such attach-ment, or both. As shown in the embodiment illustrated inFig. 5, the present invention, in this aspect, includes animpeller support having a base, substantially perpendic-ular to a shaft upon which the impeller rotates, having afirst portion 534 of average thickness sufficient to ade-quately support the impeller shaft, and a second, periph-eral portion 536 thinner than the first portion for facilitatingattachment to the bag. The first portion thickness is de-fined as the overall thickness cross-section taken up bythe first portion at any point and, where the first portionincludes a ribbed or other structure including variousthicknesses, the thickness for purposes of this discussionis defined as the thickest portion. The second, peripheralportion, in one embodiment, defines a composition sim-

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ilar to or essentially identical to that of the collapsible bag,and is provided in a thickness similar to that of the col-lapsible bag. In other embodiments, the second, periph-eral portion is formed by a composition different than thatof the collapsible bag. For instance, in some embodi-ments, the first portion is formed in low density polyeth-ylene, and the second portion is formed in high densitypolyethylene, polypropylene, silicone, polycarbonate,and/or polymethacrylate.[0090] The thickness of the peripheral portion of thesupport and the thickness of the walls of collapsible bag540, prior to attachment, may differ by no more than100%, or by no more than 80%, 60%, 40%, 20%, or 10%in other embodiments (e.g., as a percentage of the great-er thickness between the walls of the bag and the pe-ripheral portion). This aspect of the disclosure involves,in part, the discovery that where the thickness of the pe-ripheral portion of the impeller support and the thicknessof the disposable bag (at least the portion attachable tothe impeller support) are made of similar (or compatible)materials and are of similar thickness, then joining of oneto the other can be facilitated easily, reproducibly, andwith a product that is free of significant irregularity andthickness in the transition of the bag to the impeller sup-port attachment portion. Thus, one aspect of the disclo-sure involves the product of attachment of a collapsiblebag and an impeller support each as defined above, andin another aspect involves a kit including an impeller sup-port and a collapsible bag prior to attachment. As de-scribed herein, joining of the bag and the support can beperformed by any suitable method including, for example,molding (e.g., as described above in connection withFigs. 2A-2C) and welding (e.g., ultrasonic or heat weld-ing).[0091] Another aspect of the disclosure involves im-pellers with replaceable blades. Fig. 6 illustrates an im-peller 570 according to one embodiment of this aspectof the disclosure which includes a hub 572 which canhave a generally circular outer perimeter and may includea center passage 576 before within which the impellershaft or post (not illustrated) resides. Hub 572 includesone or more slots 578 within which one or more impellerblades 580 can, in some embodiments, be replaceablyinserted. As illustrated, one slot 578 is shown not con-taining a blade and one slot 578 is shown containing animpeller blade. The blade and blade slots are illustratedvery schematically and, of course, those of ordinary skillin the art will recognize that a variety of different sizes,shapes, and pitches of blades and slots can be selectedby those of ordinary skill in the art for a variety of mixingpurposes described herein and known in the art. Blades580 can be positioned and held within slots 578 securelyenough for suitable use and accordance with the inven-tion by any number of techniques including, for example,friction fitting, press fitting, detent mechanism, a clippingand clip release arrangement, fastening with screws,pegs, clamps, or the like, welding (e.g., heat and ultra-sonic welding), and use of adhesives.

[0092] The replaceable blade arrangement of the in-vention as illustrated in Fig. 6 provides the advantage inthat different blades can be used with a single hub in amixing/rotating arrangement so that the arrangement canbe used for different purposes or involving different rota-tional speed, torque, mixing profile, or the like. For ex-ample, blades of a first size or pitch can be replaced withblades of a second size or pitch to create greater or lessersheer, aeration, mixing or the like as would be understoodby those of ordinary skill in the art. While replaceableblades (e.g., airplane propeller blades) are known in dif-ferent fields, replaceable blades in a collapsible bag ar-rangement such as that of the present invention wouldnot have been expected to have been found based uponknowledge in the art because such bags typically wereused only for mixing media containing cells which, toavoid being lysed, must be stirred below a threshold ofsheer, or for media containing other materials which cantolerate much higher sheer. In this aspect of the presentinvention, collapsible bag arrangements can be preparedwith multiple blades and provided for use with either orboth of two or more mixing profiles.[0093] In one aspect, the impeller (in some embodi-ments, via magnetic coupling of the drive head to theimpeller) is driven by a motor able to reverse its directionof rotation and/or to be finely tuned with respect to rota-tional speed. Reversal of direction of spin is not found orsuggested by the known art but provides significant ad-vantage as recognized by the present invention in termsof variety of aeration/sparger profiles, or the like. Finetuning of impeller speed has been determined accordingto the present invention to allow for a precise and con-trollable degree and/or balance of aeration/sparging,sheer, or the like, which has been determined to be quiteuseful in connection with various media for mixture, es-pecially those including cells. This embodiment of theinvention allows for reproducible and controllable adjust-ment of rotational speed of the impeller that amounts ofplus or minus 5% or less through a range of rotationalspeeds of between 10% and 90% of total maximum im-peller rotational speed. In other embodiments, rotationaltuning of 4%, 3%, 2%, or 1% of this speed is facilitated.In one arrangement, these aspects are realized by useof a servo motor.[0094] In another aspect, the present disclosure in-cludes a system that can reduce foam produced withina container or reduce the amount of foam contained in ahead space of a container. In some cases, sensors and/orcontrollers may also be used to monitor and/or controlfoaming. As a specific, non-limiting example, a foam sen-sor may be inserted into a head space. In one embodi-ment, the foam sensor may include two or more foamprobes with a voltage potential between them that is ableto activate antifoaming (via a control system), for exam-ple, the addition of a chemical antifoaming agent, e.g.,via a pump. In another embodiment, a foam sensor in-cludes a first portion including a probe and a second por-tion including a wall of a reusable support structure (or a

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wall of a container) that can detect a change of electricalcurrent flow between the probe and the wall (e.g., due tothe presence of the foam). In another embodiment, afoam sensor uses an amperage draw of an external motorto detect foam in a container. In such an arrangement,the current may increase when the foam contacts an im-peller driven by the motor. In yet another embodiment,the foam sensor can activate a mechanical antifoamingdevice via a control system. As another specific non-lim-iting example, mechanical foam control may be achieved,for example, via a mounting impeller in head of a con-tainer, turned upside down, optionally with fritted or non-fritted exhaust air exit elements to enhance foam break-age. Examples of impellers that can be used in an anti-foaming system are described above in connection withFigs. 4A-8.[0095] More generally, in some embodiments, a deviceable to reduce or eliminate foam may be associated witha vessel, for example, within a container of the vessel.The antifoaming device may be able to reduce or elimi-nate foaming, for example, prior to foam accumulating inan exhaust or an outlet, plugging of filters, or the like.The antifoaming system may be run continuously, peri-odically, or in some cases, in response to certain events,e.g., within a bioreactor system and/or within the contain-er. For example, the antifoaming device may include oneor more sensors and a control system which is able tomonitor foaming and act to reduce or eliminate the foam-ing.[0096] In various embodiments, the antifoaming de-vice may include a control system, which can include oneor more foam sensors, for example, within a container.The foam sensor may be positioned in any place able todetect foaming and/or a consequence of foaming, for ex-ample, the alteration of pressure within an outlet of thecontainer. For instance, a foam sensor may be positionedin a head space within the container, in an outlet port, inan exit line (e.g., in an exit air line, and/or upstream ofan exit air filter), or the like. Detection of foam in one ormore of the foam sensors may cause the control systemto enact antifoam measures, as discussed below. In an-other set of embodiments, a sensor that detects foam orthe effects of foam in a container may be a pressuresensor that detects overpressure (e.g., due to cloggingby the foam), which may be used to determine the degreeof foaming. For example, in one embodiment, foam de-tection may occur by a pressure sensor in a head spacewithin a container, or within an outlet port.[0097] Antifoaming techniques include, but are not lim-ited to, a lowering of gas flow rate to the container and/orshut of gassing entirely, the addition of chemical anti-foaming agents, which are known to those of ordinaryskill in the art (e.g., via an external pump), a mechanicalfoam breaker mounted in head space, and/or the lower-ing or ceasing of an agitation rate of a mixer in the con-tainer.[0098] In one set of embodiments, a mechanical anti-foaming system is used to reduce or eliminate foaming.

The mechanical antifoaming system can be positionedin any suitable location within the bioreactor system suchas within the container. For instance, a mechanical anti-foaming system may be fitted to the inside head spaceof a container of the present invention, or to an outlet port(which allows air and/or foam to exit the container). Themechanical antifoaming system may have any suitablestructure able to reduce and/or eliminate foam. For in-stance, in one embodiment, the mechanical foam break-er includes one or more stainless steel plates or conesmounted on a hollow rotating shaft that penetrates thecontainer. The shaft can be rotated by an external motor(e.g., a magnetically-operated motor) or other suitableapparatus. In some cases, a hollow motor shaft may beused, which may also provide a passage for gases exitingthe container in some cases. In other cases, impellerssuch as the ones shown in Figs. 4A-8 can be used.[0099] In certain instances, a shaft associated with amechanical antifoaming device is positioned between theinside and outside of the container. The location wherethe shaft exits the container may be maintained in a sterilecondition. For instance, internal and/or external rotatingseals may be used to maintain a sterile seal, and/or livehot steam may be used to facilitate maintenance of thesterile seal. By maintaining such a sterile seal, contam-ination caused by the shaft, e.g., from the external envi-ronment, from the exiting gases, etc., may be reducedor avoided.[0100] Without wishing to be bound by any theory, it isbelieved that, as gas and/or foam passes through a me-chanical antifoaming device, for example, between spin-ning impellers, plates or cones, a centrifugal force is ap-plied to the foam which overcomes stabilizing surfacetension forces and can result in collapse of the foam bub-ble. The fluid from the collapsed bubble may thus beejected out and down to the container. Thus, the exitinggases may be at least substantially free of foam. Themechanical antifoaming device can be left running at alltimes or activated as needed via a sensor, e.g., via asensor mounted in the head space of the container, aspreviously described.[0101] Referring now to Fig. 7, one embodiment ofsuch a mechanical antifoaming system is shown. In thisfigure, a foam 602 is produced, e.g., during a chemical,biochemical, and/or a biological reaction within a con-tainer 608 (e.g., a rigid container or a collapsible bag).In this example, foaming is detected by a foaming sensor611 in electrical communication with a control system,which then causes an antifoaming device to activate.Here, the antifoaming device includes an external drivemotor 614, which rotates a shaft 617 connected to vanes622. Shaft 617 penetrates container 608 to reach its in-terior. The foam 602 is then disrupted as it is passedbetween spinning vanes 622. In addition, in this example,shaft 617 is hollow, and gases can pass through the shaft,exiting container 608, as indicated by arrows 619, pass-ing through conduits 616 and optional filter 617.[0102] In another embodiment, a mechanical anti-

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foaming device of the invention does not penetrate thecontainer. Thus, the mechanical antifoaming device maylack a rotating shaft or a hollow shaft, and/or may lack aseal. In addition, as there is no penetration of the con-tainer by such a mechanical antifoaming device, theremay be no need to maintain the mechanical antifoamingdevice in a sterile condition, e.g., using internal and/orexternal rotating seals, live hot steam, or the like. Sucha system may be formed, for instance, from steel, stain-less steel or inexpensive hard plastic for installation incollapsible or disposable bags or other containers de-scribed herein.[0103] Referring now to Fig. 8, a non-limiting exampleof such a mechanical antifoaming system is shown. Inthis figure, within container 608, foam 602 is produced,e.g., during a chemical and/or a biological reaction withinthe container. The foaming is detected by a foaming sen-sor 611, and a control system then causes an antifoamingdevice to activate. In this particular example, the anti-foaming device includes an external drive motor 614,which includes a spinning external magnetic drive head630. The lack of a penetrating shaft allows container 608to be quickly removed from motor 614. Inside container608, an internal magnetic hub 632 rotates as a result ofthe rotation of external magnetic drive head 632. Thiscauses rotation of blades 638 attached to internal mag-netic hub 632 (e.g., about a single axis), which thenbreaks down the foam. In this example, gases do notleave via a hollow shaft (as in Fig. 7), but instead passthrough an outlet port 240 (which may be hollow, containa porous frit element, etc.), exiting container 608, as in-dicated by arrows 619, passing through conduits 616 andfilter 617.[0104] Although Figs. 7 and 8 show mechanical anti-foaming devices that are positioned at or near a top por-tion of a container, in other embodiments, the devicescan be positioned at any suitable location within a con-tainer, for example, near the center portion of a container.This can be achieved, for example, by extending thelength of a shaft which supports an impeller, or by anyother suitable configuration. The impeller can also belowered or risen depending on the levels of liquid andfoam in the container. Additionally, antifoaming devicesmay include more than one impeller in some instances.For example, a first impeller coupled to a shaft may belocated near a top portion of the container and a secondimpeller coupled to the shaft may be positioned near thecenter of container. An antifoaming system of a vesselmay be disposable or intended for a single use (e.g.,along with the container), in some cases.[0105] The impeller systems described herein may al-low the system to mix fluids, solids, or foams of any type.For example, fluids inside the container may be mixedto provide distribution of nutrients and dissolved gasesfor cell growth applications. The same disposable con-tainer may be used for mixing buffers and media or othersolutions in which a disposable product contact surfaceis desirable. This may also include applications in which

the vessel is not required to be sterile or maintain sterility.Moreover, embodiments described herein enable thecontainer holding the fluids/mixtures/gases to be re-moved and discarded from the reusable support structuresuch that the reusable support structure is not soiled bythe fluids that are mixed in the container. Thus, the re-usable support structure need not to be cleaned or ster-ilized after every use.[0106] Another aspect of the disclosure includes mul-tiple spargers (including sparging elements) that may bedimensioned for connection to different sources of gasand/or which may be independently controlled. The typeof gas, number of spargers, and types and configurationsof spargers used in a bioreactor system or a biochemi-cal/chemical reaction system may depend, in part, on theparticular process to be carried out (e.g., an aerobic ver-sus anaerobic reaction), the removal of any toxic byprod-ucts from the liquid, the control of pH of a reaction, etc.As described in more detail below in connection with cer-tain embodiments described herein, a system may in-clude separate spargers for different gases which mayhave different functions in carrying out, for example, achemical, biochemical and/or biological reaction. For in-stance, a bioreactor system for cell cultivation may in-clude different types of gases such as a "dissolved oxy-gen (DO) control gas" for controlling the amount of dis-solved oxygen in the culture fluid, a "strip gas" for con-trolling the amount of toxic byproducts in the culture fluid,and a "pH control gas" for controlling the pH of the culturefluid. Each type of gas may be introduced into the cultureusing different spargers that can be independently oper-ated and controlled. Advantageously, such a system mayprovide faster process control and less process controlvariability (compared to, for example, certain systemsthat combine a DO control gas, strip gas, and pH controlgas into one gas stream introduced into a reactor). Chem-ical, biochemical and/or biological reactions carried outin bioreactor systems described herein may also requirelower consumption of gas which can save money on ex-pensive gases, and/or less total gas flow rate (e.g., for astrip gas), which can reduce foam generation and/or re-duce the size of inlet gas sterile filters required.[0107] In some embodiments, vessels described here-in are a part of a bioreactor system. In bioreactors usedfor certain types of cell cultivation, cells may require nu-trients such as sugars, a nitrogen source (such as am-monia (NH3) or amino acids), various salts, trace metalsand oxygen to grow and divide. Like the other nutrients,even and uniform distribution of oxygen throughout thereactor may be essential to provide uniform cell growth.Poor distribution of oxygen can create pockets of cellsdeprived of oxygen, leading to slower growth, alterationof the cell metabolism or even cell death. In certain ap-plications where the cells are engineered to produce abioproduct, oxygen deprivation can have a sever affecton the quantity and quality of bioproduct formation. Theamount of nutrients available to cells at any one timedepends in part on the nutrient concentration in the fluid.

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Sugars, nitrogen sources, salts, and trace metals maybe soluble in fluid and, therefore, may be in excess andreadily available to the cells. Oxygen, on the other hand,may be relatively poorly soluble or "dissolved" in water.In addition, the presence of salts plus the elevated tem-perature necessary to grow cells may further reduce dis-solved oxygen concentration. To compensate, a rapiddissolved oxygen sensing system, constant and steadytransfer of oxygen into the fluid (e.g., using one or morespargers as described herein), combined with rapid andeven distribution in the bioreactor may be used to reduceor prevent oxygen starvation.[0108] Since oxygen transfer from the gas bubbles en-tering the fluid of the culture may be an important controlparameter, the time constant of responsiveness of thegas delivery system may also be important. In certainembodiments, as cell population density increases, theresponse rate of the gassing system to supply oxygenenriched DO control gas may become increasingly im-portant. Accordingly, in some embodiments, systems de-scribed herein include one or more sensors such as aDO sensor which detects the need for more oxygen (orother gas), a gas controller, and one or more spargerswhich can be signaled to enrich the culture with extraoxygen using, for example, a N2/O2/air control gas. Sincedelay time (e.g., several minutes) for this enriched gasto reach the reactor can result in a drop in DO which canlead to oxygen starvation, systems described herein mayinclude a control feedback loop between the sensor(s),gas controller, and sparger(s). Thus, responsive, andeven supply and distribution of oxygen-bearing controlgas (e.g., a N2/O2/air mix) may be provided for controlled,predictable cell growth and bioproduct formation. Sys-tems described herein allowing independent control ofspargers and/or gas compositions may be advantageouscompared to systems that require gases to be flushedout before sparging a different gas into the container.[0109] In addition, since compressed air and oxygenmay be expensive to supply to the reactor, a system thatprovides just enough air enriched with just enough oxy-gen such that the bubbles are not lost to the head spaceof the container (and lost out through the exhaust line)may be implemented. This can be performed, for exam-ple, by controlling the amount and flow rate of a controlgas independently of other gases used in the system(e.g., a strip gas and/or a pH control gas).[0110] Without wishing to be bound by any theory, it isbelieved that the rate of oxygen transfer into the biore-actor fluid from air, pure oxygen or a gas mixture is directlyrelated to the amount of total surface area of the bubblesin the fluid. Hence, larger bubbles provide less total sur-face area than a fine mist of very small bubbles. For thisreason, in certain embodiments of the invention, a controlgas may be provided through microporous spargers tocreate very small bubbles. A microporous sparger mayinclude apertures having a size (e.g., average diameter)of, for example, less than less than 500 microns, lessthan 200 microns, less than 100 microns, less than 60

microns, less than 50 microns, less than 40 microns, lessthan 30 microns, less than 20 microns, less than 10 mi-crons, less than 3 microns, less than about 1 micron, orless than 0.1 microns. In certain embodiments, micropo-rous spargers have an aperture size between 0.1 and100 microns. Of course, spargers having larger aperturesizes may also be used. For instance, a sparger mayhave an aperture size between 0.1 and 10 mm. The ap-erture size may be greater than 100 microns, greaterthan 200 microns, greater than 500 microns, greater than1 mm, greater than 3 mm, greater than 5 mm, greaterthan 7 mm, or greater than 10 mm. The aperture mayhave any suitable cross-sectional shape (e.g., circular,oval, triangular, irregular, square or rectangular, or thelike). Spargers having combinations of aperture sizes canbe incorporated into vessels described herein.[0111] Additionally, good cell growth and controlledmetabolism may be dependent upon removal of toxic by-products of cell growth, such as, for example, carbondioxide, ammonia and volatile organic acids. Carbon di-oxide may be highly soluble in water, which can exacer-bate its toxic effect on cells. These byproducts can be"stripped" out of the culture fluid by gassing the cultureusing a strip gas. Accordingly, even distribution of stripgas and strip gas that is introduced at a flow rate suffi-ciently high enough for bubbles to escape out of the cul-ture (and out the exhaust vent, for example) may be im-portant for cell growth and/or bioproduct production.These parameters may be controlled independently ofother gases used in the system (e.g., a control gas and/ora pH control gas) using a separate sparger for the stripgas.[0112] In some instances, a strip gas is introduced intoa container using a sparger having an aperture size be-tween 0.1 and 10 mm. For example, the aperture sizemay be greater than 100 microns, greater than 200 mi-crons, greater than 500 microns, greater than 1 mm,greater than 3 mm, greater than 5 mm, greater than 7mm, or greater than 10 mm. These aperture sizes canallow relatively larger bubbles to pass through the liquidof the container, which can strip any toxic byproducts outof the liquid without creating large amounts of foam inthe head space of the container.[0113] In certain embodiments, a pH control gas isused to control the pH of the fluid in a bioreactor system.For example, carbon dioxide may be used to increasesolution pH and ammonia may be used to decrease so-lution pH. In one embodiment, a pH control gas may in-clude a combination of carbon dioxide, ammonia, or othergases to control (e.g., increase or decrease) pH. In an-other embodiment, the pH of a reaction fluid is controlledby a first sparger containing an agent that increases pH(e.g., CO2) and a second sparger containing an agentthat decreases pH (e.g., NH3).[0114] One or more pH control gases may be addedto a container of the bioreactor system upon signals froma pH control sensor associated with the system. The pHcontrol gases may be operated independently and with-

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out interference by oxygen demand (e.g., a DO controlgas) or strip gas systems. A pH control gas may be in-troduced into a container using spargers having aper-tures of various sizes.[0115] In other embodiments, cells that are normallygrown without oxygen (e.g., anaerobic reactions) orwhich are even sensitive to oxygen require removal ofoxygen from the culture. Even and controlled distributionof nitrogen gas in these cultures may be used to controlproper cell growth and product formation.[0116] As mentioned, in some embodiments of thepresent disclosure, gases such as air, CO2, O2, N2, NH3,and/or dissolved oxygen may be sparged into the con-tainer. In some cases, the sparging can be controlled,for instance, such that the sparging can be rapidly acti-vated or altered as needed. Multiple spargers may beused in some cases. For example, in one embodiment,different gas compositions may each be introduced intothe container using multiple spargers, e.g., a first spargerfor a first gas composition, a second sparger for a secondgas composition, a third sparger for a third gas compo-sition, etc. The gases may differ in composition and/or inconcentration. As a specific example, a first gas compo-sition may include air with 5% CO2, and a second gascomposition may include air with 10% CO2; in anotherexample, a first gas composition may include O2, and asecond gas composition may include N2; in yet anotherexample, a first gas composition may include a controlgas, a second gas composition may include a strip gas,and a third gas composition may include a pH controlgas. Of course, other combinations of gases are alsopossible. In some cases, multiple spargers may be usefulto allow faster responses, e.g., as the gas compositionbeing introduced into the container may be rapidlychanged by activating different spargers, e.g., singlyand/or in combination. As a specific example, the gasbeing introduced into a container can be rapidly switchedfrom a first gas (via a first sparger) to a second gas (viaa second sparger), and/or to a combination of the firstand second gas, or a combination of the second gas anda third gas, etc. The flow rates of each gas can also bechanged independently of one another. (In contrast, witha single sparger, a change in composition requires thatthe new composition reach the sparger before being in-troduced into the container.) Moreover, the use of multi-ple spargers can allow customization of the type of sparg-er for a particular type of gas, e.g., a strip gas, DO controlgas, pH control gas, air, CO2, O2, N2, NH3, or any othersuitable gas, if desired.[0117] Sparging may be run continuously, periodically,or in some cases, in response to certain events, e.g.,within a bioreactor system and/or within the container.For example, as mentioned, the spargers may be con-nected to one or more sensors and a control systemwhich is able to monitor the amount of sparging, the de-gree of foaming, the amount or concentration of a sub-stance in the container, and respond by initiating, reduc-ing, or increasing the degree of sparging of one or more

composition(s) of gases.[0118] In one particular embodiment, a vessel (e.g., aspart of a reactor system for performing a biological, bio-chemical or chemical reaction) is configured to containa volume of liquid and includes a container (e.g., a col-lapsible bag) having a volume of at least 2 liters (or anyother suitable volume) to contain the volume of the liquid.The vessel may optionally include a support structure forsurrounding and containing the container. Additionally,the vessel includes a first sparger connected or dimen-sioned to be connected to a source of a first gas compo-sition in fluid communication with the container, and asecond sparger connected or dimensioned to be con-nected to a source of a second gas composition differentfrom the first gas composition in fluid communication withthe container. The vessel further comprises a control sys-tem operatively associated with the first and secondspargers and configured to operate the spargers inde-pendently of each other. Of course, third, fourth, fifth, orgreater numbers of spargers can be included (e.g., great-er than 10, or greater than 20 spargers), depending on,for example, the size of the container. In some embodi-ments, the vessel further comprises a mixing system in-cluding an impeller and a base plate, wherein the firstand/or second spargers is associated with the base plate.In one particular embodiment, the first gas compositioncomprises air and the second gas composition comprisesair supplemented with O2 and N2. If additional spargersare included, the spargers can be connected to a sourceof gas comprising N2, CO2, NH3 and/or any other suitablegas.[0119] In another exemplary embodiment, a vesselconfigured to contain a volume of liquid comprises a con-tainer (e.g., a collapsible bag) to contain the liquid, andoptionally, a support structure for surrounding and con-taining the collapsible bag. The vessel includes a firstsparger connected to the container, the first sparger hav-ing a first aperture size, wherein at least a portion of thefirst sparger is dimensioned to be connected to a sourceof a first gas composition. The vessel also includes asecond sparger connected to the container, the secondsparger having a second aperture size, wherein at leasta portion of the second sparger is dimensioned to beconnected to a source of a second gas composition. Thesecond gas composition may have the same or a differentcomposition than the first gas composition. In some em-bodiments, the vessel is part of a bioreactor system; or,the vessel may be a part of a biochemical/chemical re-action system, or a mixing system. The vessel may in-clude a control system operatively associated with thefirst and second spargers and may be configured to op-erate the spargers (or gases associated therewith) inde-pendently of each other. The vessel may include anysuitable number of spargers (e.g., greater than 10 orgreater than 20 spargers), and the container may haveany suitable volume (e.g., at least 2, 10, 20, 40, or 100liters). The first and/or second gas composition(s) mayinclude, for example, N2, O2, CO2, NH3, or air. For ex-

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ample, in one instance, the first gas comprises air andthe second gas comprises air supplemented with O2 andN2. The first aperture size may be larger than the secondaperture size. For instance, the first aperture size maybe between 0.1 and 10 mm, and the second aperturesize may be between 0.1 and 100 microns.[0120] Apertures associated with spargers can beformed in any suitable material. For instance, in one em-bodiment, a porous polymeric material is used as a sparg-ing element to allow transport of gas from one side toanother side of the material. Apertures can also beformed in other materials such as metals, ceramics, pol-ymers, and/or combinations thereof. Materials havingpores or apertures can have any suitable configuration.For example, the materials may be knitted, woven, orused to form meshes or other porous elements. The el-ements may be in the form of sheets, films, and blocks,for example, and may have any suitable dimension. Insome cases, such elements are incorporated with impel-lers or impeller supports, e.g., as illustrated in Fig. 5. Theelements can be positioned and held within regions ofthe impeller or impeller support securely enough for suit-able use and accordance with the invention by anynumber of techniques including, for example, friction fit-ting, press fitting, detent mechanism, a clipping and cliprelease arrangement, fastening with screws, pegs,clamps, or the like, welding (e.g., heat and ultrasonicwelding), and use of adhesives. In other embodiments,portions of the impeller and/or impeller support can befabricated directly with pores or apertures that can allowfluids to flow therethrough.[0121] The vessel may optionally include one or moresensors in electrical communication with the control sys-tem for determining an amount or concentration of a gas(e.g., O2, N2, CO2, NH3, a bi-product of a reaction) in thecontainer. Additionally and/or alternatively, the vesselmay include a sensor in electrical communication withthe control system for determining a pH of a liquid in thecontainer, or an amount or level of a foam in the bag.[0122] As mentioned, control systems and feedbackloops may be used to control the degree of sparging inone embodiment, or degree of mixing, or activity of anantifoaming system in other embodiments. One exampleof such a control and feedback process is shown in theembodiment illustrated in Fig. 9. System 700 may includea first sensor 702 (e.g., for detecting the amount and/orconcentration of CO2 of a liquid in the container) and asecond sensor 704 (e.g., for detecting the amount and/orconcentration of O2 of a liquid in the container). Aftercalibrating the sensors, reagents may be added to a con-tainer 708 and a fluidic manipulation process, such asmixing or performing a biological, chemical, or biochem-ical reaction, may be take place. The amount of a gassuch as O2 and CO2 may vary in the liquid of the containeras the process proceeds. For example, if a biologicalreaction involving cells takes place, the cells may con-sume O2 and form CO2 over time, which may vary de-pending on the growth stage of the cells. Thus, the

amount and/or concentration of gases can be determinedby the sensors (e.g., as a function of time), and signals712 and 714 related to the amounts and/or concentra-tions of the gases can be sent to a control system 720.The control system may include recorded parameters724, such as threshold levels of one or more gases thatcan inputted by a user prior to or during the reaction. Forexample, a parameter may include a certain thresholdlevel of CO2 in the liquid before a sparger is activated toreduce the amount of CO2 using a strip gas. Accordingly,a signal may be sent from the control system to activatea component 732, such as a valve connected to a sourceof a strip gas used to reduce the amount of CO2. As thestrip gas is introduced into container 708, the amountand/or concentration of CO2 may decrease, which canbe measured by 712 and signals sent to the control sys-tem. When the amount and/or concentration of CO2 de-creases to a certain level, the control system can loweror deactivate the amount of CO2 being introduced intothe container, thereby completing the feedback loop. Asimilar process can take place independently of the proc-ess described above using second sensor 714, whichmay measure, for example, a second gas, a pH, or anamount of a foam in a head space of the container.[0123] In another aspect, a bubble column or airlift sys-tem (utilizing bubbles of air or other gas) may be usedwith the disposable bioreactor bag. Such a system mayprovide a mixing force by the addition of gas (e.g., air)near the bottom of the reactor. Here, the rising gas bubbleand the lower density of gas-saturated liquid rise, dis-placing gas-poor liquid which falls, providing top-to-bot-tom circulation. The path of rising liquid can be guided,for example, using dividers inside the chamber of thebag. For instance, using a sheet of plastic which bisectsthe interior of the bioreactor bag, e.g., vertically, with agap at the top and the bottom. Gas may be added on oneside of the divider, causing the gas and gas-rich liquid torise on one side, cross over the top of the barrier sheet,and descend on the other side, passing under the dividerto return to the gas-addition point. In addition, such abubble column/air-lift mixing system and method may becombined with any of the other mixing systems describedherein.[0124] In one aspect, a bioreactor system as describedherein includes an enclosed resin loading/column pack-ing system. Typically, column packing typically may beaccomplished in a clean room with open carboys con-taining the resin which is manually mixed while the resinslurry is pumped onto the column. In one embodiment,however, a container such as a flexible container is load-ed with chromatography resin which is slurried by an ag-itator while the slurry is pumped into a column.[0125] In certain chemical, biochemical and/or biolog-ical processes requiring light, a bioreactor system de-scribed herein may include direct, indirect and/or piped-in lighting, e.g., using fiber-optics, according to anotheraspect of the invention. Any suitable light source may beused. Such bioreactor systems may be useful for

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processing, for example, plant cells, e.g., to activate pho-tosynthesis. In one particular embodiment, a phospho-rescent flexible container is used to provide light, e.g.,for growth of plant cells.[0126] While several embodiments of the present dis-closure have been described and illustrated herein, thoseof ordinary skill in the art will readily envision a variety ofother means and/or structures for performing the func-tions and/or obtaining the results and/or one or more ofthe advantages described herein, and each of such var-iations and/or modifications is deemed to be within thescope of the present invention. More generally, thoseskilled in the art will readily appreciate that all parameters,dimensions, materials, and configurations describedherein are meant to be exemplary and that the actualparameters, dimensions, materials, and/or configura-tions will depend upon the specific application or appli-cations for which the teachings of the present inventionis/are used. Those skilled in the art will recognize, or beable to ascertain using no more than routine experimen-tation, many equivalents to the specific embodiments ofthe invention described herein. It is, therefore, to be un-derstood that the foregoing embodiments are presentedby way of example only and that, within the scope of theappended claims and equivalents thereto, the inventionmay be practiced otherwise than as specifically de-scribed and claimed. The present invention is directedto each individual feature, system, article, material, kit,and/or method described herein. In addition, any combi-nation of two or more such features, systems, articles,materials, kits, and/or methods, if such features, sys-tems, articles, materials, kits, and/or methods are not mu-tually inconsistent, is included within the scope of thepresent invention.[0127] All definitions, as defined and used herein,should be understood to control over dictionary defini-tions, definitions in documents incorporated by refer-ence, and/or ordinary meanings of the defined terms.[0128] The indefinite articles "a" and "an," as usedherein in the specification and in the claims, unless clearlyindicated to the contrary, should be understood to mean"at least one."[0129] It should also be understood that, unless clearlyindicated to the contrary, in any methods claimed hereinthat include more than one step or act, the order of thesteps or acts of the method is not necessarily limited tothe order in which the steps or acts of the method arerecited.[0130] In the claims, as well as in the specificationabove, all transitional phrases such as "comprising," "in-cluding," "carrying," "having," "containing," "involving,""holding," "composed of," and the like are to be under-stood to be open-ended, i.e., to mean including but notlimited to. Only the transitional phrases "consisting of"and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively.

Claims

1. A vessel configured to contain a volume of liquidcomprising:

a container able to contain the volume of liquid,wherein the container is a collapsible bag;a reusable support structure surrounding andcontaining the collapsible bag; anda magnetically-driven antifoaming device, whichis positioned in a head space of the containerwhen the container contains the volume of liquid,the antifoaming device being configured and ar-ranged to break up foam in the head space dur-ing rotation of at least a portion of the antifoam-ing device, wherein the antifoaming device com-prises an impeller that is magnetically rotatedabout a single axis.

2. A vessel as in claim 1, wherein the impeller is con-nected to a top portion of the container.

3. A vessel as in one of the preceding claims, the vesselfurther comprising a sensor and a control system,the control system being operatively associated withthe magnetically-driven antifoaming device and thesensor.

4. A vessel as in claim 3, wherein the sensor is a foamsensor.

5. A vessel as in claim 3 or 4, wherein the sensor com-prises a first portion including a probe and a secondportion including a wall of the vessel.

6. A vessel as in claim 5, wherein the antifoaming de-vice is regulated by the control system via detectionof electrical current flow between the probe and thewall.

7. A vessel as in claim 1, further comprising:

a pressure sensor for determining a pressure inthe collapsible bag, the pressure sensor beingin fluid communication with the collapsible bag;and a control system operatively associated withthe pressure sensor and the antifoaming device,wherein the control system regulates the anti-foaming device upon receipt of a signal from thepressure sensor.

Patentansprüche

1. Gefäß, das zur Aufnahme eines Flüssigkeitsvolu-mens konfiguriert ist, umfassend :

einen Behälter, der in der Lage ist, ein Flüssig-

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keitsvolumen zu enthalten, wobei der Behälterein zusammenfaltbarer Beutel ist; undeine wiederverwendbare Trägerstruktur, dieden zusammenfaltbaren Beutel umgibt und ent-hält; undeine magnetisch angetriebene Antischaumvor-richtung, die in einem oberen Raum des Behäl-ters positioniert ist, wenn der Behälter das Flüs-sigkeitsvolumen enthält, wobei die Antischaum-vorrichtung derart konfiguriert und angeordnetist, dass sie Schaum in dem oberen Bereich auf-bricht während sich mindestens ein Teil der An-tischaumvorrichtung dreht, wobei die Anti-schaumvorrichtung ein Laufrad umfasst, dasmagnetisch um eine einzelne Achse gedrehtwird.

2. Gefäß nach Anspruch 1, wobei das Laufrad mit ei-nem oberen Abschnitt des Behälters verbunden ist.

3. Gefäß nach einem der vorstehenden Ansprüche,wobei das Gefäß ferner einen Sensor und ein Steu-ersystem umfasst, wobei das Steuersystem operativmit der magnetisch angetriebenen Antischaumvor-richtung und dem Sensor zusammenhängt.

4. Gefäß nach Anspruch 3, wobei der Sensor einSchaumsensor ist.

5. Gefäß nach Anspruch 3 oder 4, wobei der Sensoreinen ersten Abschnitt umfasst, der eine Sonde be-inhaltet und einen zweiten Abschnitt, der eine Wanddes Gefäßes beinhaltet.

6. Gefäß nach Anspruch 5, wobei die Antischaumvor-richtung durch das Steuersystem über die Erfassungeines elektrischen Stromflusses zwischen der Son-de und der Wand gesteuert wird.

7. Gefäß nach Anspruch 1, ferner umfassend:

einen Drucksensor zur Bestimmung einesDrucks im zusammenfaltbaren Beutel, wobeider Drucksensor in Fluidkommunikation mitdem zusammenfaltbaren Beutel ist;und ein Steuersystem, das operativ mit demDrucksensor und der Antischaumvorrichtungverbunden ist, wobei das Steuersystem die An-tischaumvorrichtung auf den Empfang eines Si-gnals vom Drucksensor hin reguliert.

Revendications

1. Cuve configurée pour contenir un volume de liquidecomprenant :

un récipient qui est à même de contenir le volu-

me de liquide, dans laquelle le récipient est unsac repliable ;une structure de support réutilisable entourantet contenant le sac repliable ; etun dispositif anti-mousse entraîné magnétique-ment, qui est positionné dans un espace de têtedu récipient lorsque le récipient contient le vo-lume de liquide, le dispositif anti-mousse étantconfiguré et agencé pour fractionner la moussede l’espace de tête au cours de la rotation d’aumoins une partie du dispositif anti-mousse, danslaquelle le dispositif anti-mousse comprend unehélice qui effectue une rotation magnétiqueautour d’un seul axe.

2. Cuve selon la revendication 1, dans laquelle l’héliceest raccordée à une partie supérieure du récipient.

3. Cuve selon l’une quelconque des revendicationsprécédentes, la cuve comprenant en outre un cap-teur et un système de commande, le système decommande étant associé en service au dispositif an-ti-mousse magnétiquement entraîné et au capteur.

4. Cuve selon la revendication 3, dans laquelle le cap-teur est un capteur de mousse.

5. Cuve selon la revendication 3 ou 4, dans laquelle lecapteur comprend une première partie comprenantune sonde et une seconde partie comprenant uneparoi de la cuve.

6. Cuve selon la revendication 5, dans laquelle le dis-positif anti-mousse est régulé par le système decommande via la détection de l’écoulement de cou-rant électrique entre la sonde et la paroi.

7. Cuve selon la revendication 1, comprenant en outre :

un capteur de pression pour déterminer unepression dans le sac repliable, le capteur depression étant en communication fluidique avecle sac repliable ;et un système de commande associé en serviceau capteur de pression et au dispositif anti-mousse, dans laquelle le système de comman-de régule le dispositif anti-mousse à la réceptiond’un signal issu du capteur de pression.

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REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the Europeanpatent document. Even though great care has been taken in compiling the references, errors or omissions cannot beexcluded and the EPO disclaims all liability in this regard.

Patent documents cited in the description

• US 4310437 A [0002]• DE 2548441 A [0002]• JP H06153902 A [0002]• US 90397707 P [0017]• US 20080213874 A [0017]• US 14712405 A [0017] [0065] [0077]

• US 20050272146 A [0017] [0065] [0077]• US 2005020083 W [0017]• WO 2005118771 A [0017]• WO LTS2005002985 A [0017]• WO 2005076093 A [0017]