WO2005068059A1 - Mixing bag with integral sparger and sensor receiver - Google Patents
Mixing bag with integral sparger and sensor receiver Download PDFInfo
- Publication number
- WO2005068059A1 WO2005068059A1 PCT/US2005/000464 US2005000464W WO2005068059A1 WO 2005068059 A1 WO2005068059 A1 WO 2005068059A1 US 2005000464 W US2005000464 W US 2005000464W WO 2005068059 A1 WO2005068059 A1 WO 2005068059A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- fluid
- bag
- receiver
- vessel
- rigid
- Prior art date
Links
- 239000012530 fluid Substances 0.000 claims abstract description 71
- 238000010364 biochemical engineering Methods 0.000 claims abstract description 17
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- -1 polyethylene Polymers 0.000 description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
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- 230000001954 sterilising effect Effects 0.000 description 2
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- 241000251468 Actinopterygii Species 0.000 description 1
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- 229910000831 Steel Inorganic materials 0.000 description 1
- RTAQQCXQSZGOHL-UHFFFAOYSA-N Titanium Chemical compound [Ti] RTAQQCXQSZGOHL-UHFFFAOYSA-N 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000002457 bidirectional effect Effects 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- UBAZGMLMVVQSCD-UHFFFAOYSA-N carbon dioxide;molecular oxygen Chemical compound O=O.O=C=O UBAZGMLMVVQSCD-UHFFFAOYSA-N 0.000 description 1
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- 239000000565 sealant Substances 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
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Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/50—Mixing receptacles
- B01F35/513—Flexible receptacles, e.g. bags supported by rigid containers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
- B01F23/20—Mixing gases with liquids
- B01F23/23—Mixing gases with liquids by introducing gases into liquid media, e.g. for producing aerated liquids
- B01F23/231—Mixing gases with liquids by introducing gases into liquid media, e.g. for producing aerated liquids by bubbling
- B01F23/23105—Arrangement or manipulation of the gas bubbling devices
- B01F23/2312—Diffusers
- B01F23/23121—Diffusers having injection means, e.g. nozzles with circumferential outlet
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
- B01F23/20—Mixing gases with liquids
- B01F23/23—Mixing gases with liquids by introducing gases into liquid media, e.g. for producing aerated liquids
- B01F23/231—Mixing gases with liquids by introducing gases into liquid media, e.g. for producing aerated liquids by bubbling
- B01F23/23105—Arrangement or manipulation of the gas bubbling devices
- B01F23/2312—Diffusers
- B01F23/23123—Diffusers consisting of rigid porous or perforated material
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
- B01F23/20—Mixing gases with liquids
- B01F23/23—Mixing gases with liquids by introducing gases into liquid media, e.g. for producing aerated liquids
- B01F23/233—Mixing gases with liquids by introducing gases into liquid media, e.g. for producing aerated liquids using driven stirrers with completely immersed stirring elements
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F27/00—Mixers with rotary stirring devices in fixed receptacles; Kneaders
- B01F27/80—Mixers with rotary stirring devices in fixed receptacles; Kneaders with stirrers rotating about a substantially vertical axis
- B01F27/808—Mixers with rotary stirring devices in fixed receptacles; Kneaders with stirrers rotating about a substantially vertical axis with stirrers driven from the bottom of the receptacle
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/45—Magnetic mixers; Mixers with magnetically driven stirrers
- B01F33/453—Magnetic mixers; Mixers with magnetically driven stirrers using supported or suspended stirring elements
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/45—Magnetic mixers; Mixers with magnetically driven stirrers
- B01F33/453—Magnetic mixers; Mixers with magnetically driven stirrers using supported or suspended stirring elements
- B01F33/4534—Magnetic mixers; Mixers with magnetically driven stirrers using supported or suspended stirring elements using a rod for supporting the stirring element, e.g. stirrer sliding on a rod or mounted on a rod sliding in a tube
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/20—Measuring; Control or regulation
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/20—Measuring; Control or regulation
- B01F35/21—Measuring
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/20—Measuring; Control or regulation
- B01F35/21—Measuring
- B01F35/213—Measuring of the properties of the mixtures, e.g. temperature, density or colour
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/50—Mixing receptacles
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/14—Bags
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/22—Transparent or translucent parts
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F2101/00—Mixing characterised by the nature of the mixed materials or by the application field
- B01F2101/23—Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/45—Magnetic mixers; Mixers with magnetically driven stirrers
Definitions
- the present invention relates generally to vessels in which fluids are agitated and, more particularly, to a mixing bag including an integral sparger and sensor receiver.
- a traditional bioreactor is a sterile vessel made out of stainless steel or glass with highly controlled environmental parameters including temperature, pH, oxygen concentration, CO2 concentration, which are monitored by permanent sensors built into the rigid vessel.
- the fluid in the bioreactor must also be agitated in order to maintain uniform distribution of temperature, gases and nutrients.
- agitation is typically provided by an impeller with the blades housed on the shaft connected to an external motor and introduced inside the bioreactor through the dynamic seal in an effort to maintain sterility.
- concentration of dissolved oxygen must be maintained.
- controlled introduction of other gases like carbon dioxide and nitrogen are normally necessary during bioreactor runs.
- the most efficient way of introducing gases in to bioreactor fluid is sparging, which involves forming small bubbles in the fluid. Such bubbles have large surface to volume ratio and thus can be dissolved more quickly than large size bubbles.
- porous solid materials like titanium associated with the rigid bioreactor provide sparging.
- metal sparging rings with small pre-drilled holes are permanently affixed in some rigid bioreactors.
- the bioreactors are not readily disposable and thus must be cleaned and sterilized before reuse for bioprocessing.
- the impeller, sparger, gas, temperature and pH sensors are reusable components that must be cleaned and sterilized after each batch.
- it is desirable that all the fluid touching components are only used once. This presents the challenging task of providing inexpensive fluid-touching components that can be discarded along with the bag after use. Another challenge is positioning the components of the bioreactor on the flexible bag.
- a bioreactor plastic bag which is basically thin film
- bioreactor components like impeller shafts, spargers, sensors are housed on the rigid walls of the vessel by means of threads, bolts or clamps. Obviously, this method of component attachment does not work for plastic bags. Thus, a need is identified for an improved manner of providing a mixing bag or flexible vessel with an integrated sparger and sensor(s).
- the improvement provided by the invention would be easy to implement using existing manufacturing techniques and without significant additional expense. Overall, a substantial gain in efficiency and ease of use would be realized as a result of the improvement, and would greatly expand the potential applications for which advanced mixing systems may be used, including bioprocessing.
- a vessel intended for use in bioprocessing comprises a bag capable of receiving and holding the fluid.
- the bag includes a rigid, integral sparger including means for forming bubbles from a gas supplied to the fluid in the bag.
- the sparger is welded to the bag to form a fluid-impervious seal.
- the means for forming bubbles comprises a perforated film attached to the rigid material, but may instead comprise a gas permeable film attached to the rigid material.
- the means for forming bubbles comprises a perforated piece of material (including possibly a film) attached to a face of the rigid material forming the sparger.
- the rigid material further includes a first receiver for receiving and holding a magnetic fluid-agitating element at a home location when positioned in the vessel.
- the receiver is an upstanding post
- the means for forming bubbles comprises an outer wall of the post.
- the post may also provide a centering or alignment function for any fluid-agitating element positioned in the bag, and may include an oversized head for retaining it in place.
- a vessel intended for use in bioprocessing with a fluid is disclosed.
- the vessel comprises a bag capable of receiving and holding the fluid.
- the bag includes a rigid receiver facing an interior portion.
- a transparent tube having a closed end is also provided for insertion through the receiver and in communication with the interior.
- the rigid receiver is welded to the bag to form a fluid-impervious seal.
- a bushing is also provided for holding the receiver in the tube.
- Means for sealing the tube in the receiver may also be provided to stem fluid leakage when the bag is filled.
- a fluorescent sensor element is externally attached to the closed end of the tube.
- the invention further comprises the combination of the vessel with a fiber optic cable for transmitting light to the sensor element and receiving the light reflected therefrom. Use of such a sensor arrangement allows for various characteristics of the fluid to be observed in a substantially non-invasive fashion.
- a method of manufacturing a bioreactor comprises providing a bag capable of receiving and holding a fluid.
- the method further comprises attaching to the bag an integral sparger including means for forming bubbles from a gas supplied to the fluid in the bag.
- the step of placing a magnetic fluid-agitating element in the bag may also be performed.
- a method of bioprocessing a fluid comprises providing a bag capable of receiving and holding a fluid; attaching to the bag an integral sparger including means for forming bubbles in the fluid when present; supplying gas to the sparger; and disposing of the bag, including the sparger.
- the method may further include the step of recovering the fluid from the bag.
- the sparger comprises a rigid material
- the step of attaching comprises welding the bag to the rigid material to create a fluid-impervious seal.
- the method may further comprise mixing the fluid using a magnetic fluid- agitating element.
- a method of arranging a sensor for a fluid bioprocessing operation comprises providing a bag capable of receiving and holding a fluid, and attaching to the bag a rigid receiver. A transparent tube including a closed end is then inserted in the receiver. The sensor may be positioned on or in the tube.
- the method further comprises the steps of disposing of the bag with the tube in the receiver.
- the method may further comprise inserting a fiber optic cable in an open end of the tube.
- Still another part of the method may comprise externally attaching a fluorescent sensor element to a closed end of the tube.
- Figures 1 , 1 a, lb and 1 c are a partially schematic, partially cross-sectional side views of one embodiment of a vessel in the form of a bag having a flexible portion and a rigid portion;
- Figure 2 is a partially schematic, partially cross-sectional side view showing the vessel of Figure 1 positioned in a rigid vessel, with the fluid-agitating element aligned with and levitated/rotated by an adjacent motive device;
- Figure 3 a is partially schematic, partially cross-sectional side view showing another embodiment of the vessel, including a hat or cap-shaped rigid portion having a cavity facing inwardly;
- Figure 3b is a side view similar to Figure 3a;
- Figure 4a is partially schematic, partially cross-sectional side view showing another embodiment of the vessel, including a hat or cap-shaped rigid portion having a cavity facing outwardly;
- Figure 4b is a side view similar to Figure 4a;
- Figures 5a, 5b, 6a, 6b, and 7a, 7b are each partially schematic
- the bag 10 includes a body having a flexible or non-rigid portion 12, which is illustrated schematically, and a rigid or stiff portion 14, which is shown in cross-section.
- the bag 10 may be hermetically sealed and may have one or more openings or fittings (not shown) for introducing or recovering a fluid.
- the bag 10 may be unsealed or open- ended.
- the particular geometry of the bag 10 employed normally depends on the application and is not considered critical to the invention.
- the bag 10 is capable of receiving and at least temporarily holding a fluid (which is used herein to denote any substance capable of flowing, as may include liquids, liquid suspensions, gases, gaseous suspensions, or the like, without limitation).
- the rigid portion 14 includes a first receiver 1 for receiving and holding a fluid-agitating element 18 at a home location (or expected position), when positioned in the bag 10.
- holding as used herein defines both the case where the fluid-agitating element 18 is directly held and supported by the first receiver 16 (see below) against any significant side-to-side movement (save tolerances), as well as where the first receiver 16 merely limits the fluid-agitating element to a certain degree of side-to-side movement within the bag 10.
- an opening 18a is provided in the fluid-agitating element 18 and the first receiver 16 is a post 20 projecting toward the interior of the bag 10 (see Figures la and lb).
- the post 20 is sized for receiving the fluid- agitating element 18 by extending through the opening 18a formed in the body 18b thereof (which is depicted as being annular, but not necessarily circular in cross-section).
- the size of the opening 18a is such that the fluid-agitating element 18 may freely rotate and move in the axial direction along the post 20 without contacting the outer surface thereof.
- the post 20 serving as the first receiver 16 is still considered to hold, confine, or keep the fluid-agitating element 18 at a home location or expected position within the vessel 20 by contacting the surface adjacent to the opening 18a as a result of any side-to-side movement (the boundaries of which are defined by the dimensions of the opening).
- the flexible portion 12 of the bag 10 may be made from one or more sheets of thin (e.g., having a thickness of between 0.1 and 0.2 millimeters) polyethylene film secured together to define a compartment for receiving the fluid.
- the film used is clear or translucent, although the use of opaque or colored films is also possible.
- the rigid portion 14 including the post 20 may be formed of materials, such as high density polyethylene (HDPE), ultrahigh molecular weight (UHMW) polyethylene, or like materials. Of course, these materials do have som.e inherent flexibility when used to form relatively thin components or when a moderate amount of bending force is applied thereto. Despite this flexibility, the rigid portion 14 is distinguished from the flexible portion 12, in that it generally maintains its shape under the weight of fluid introduced in the bag 10.
- the post 20 may include a portion 20a for capturing the fluid-agitating element 18 and assisting in holding it thereon.
- the portion 20a is preferably oversized and forms the head or end of the post 20.
- oversized it is meant that at least one dimension (length, width, diameter) of this portion 20a of the post 20 is greater than the corresponding dimension of the opening 18a in the fluid-agitating element 18.
- the portion 20a is shown in Figure 1 as being disc-shaped, such that it provides the head end of the post 20 with a generally T-shaped cross section.
- the oversized portion 20a is strategically positioned at a certain distance along the post 20.
- the post 20 maybe removably attached to the rigid portion 14 through the opening 18a in the fluid-agitating element 18 (such as by providing a threaded bore in the rigid portion for receiving a threaded end of the post, or as shown in Figure 1 c, a bore 14a having a groove 14b for establishing a snap-fit engagement with a corresponding projection 20b on a tapered end portion 20c of the post).
- this portion should be sufficiently thin such that it flexes or temporarily deforms to allow the fluid-agitating element 18 to pass initially (see Figure lb and note action arrow A, which demonstrates the direction of force for deforming the oversized head 20a such that it passes through the opening 18a).
- this portion 20a of the post 20 need not be oversized, as defined above, but instead may simply be sufficiently close in size to that of the opening 18a such that the fluid- agitating element 18 must be precisely aligned and register with the post 20 in order to be received or removed.
- the fluid-agitating element 18 is held in place in the vicinity of the post 20, but remains free of direct attachment.
- the first receiver 16 (post 20) confines or holds the fluid-agitating element 18 at a home location or expected position within the bag 10, it is still free to move side-to-side to some degree (which in this case is defined by the size of the opening 18a), and to move along the first receiver 16 in the axial direction (vertical, in the embodiment shown in Figure 1), as is necessary for levitation.
- the rigid portion 14 in this embodiment further includes a substantially planar peripheral flange 22.
- the flange 22 maybe any shape or size, and is preferably attached or connected directly to the bag 10 at the interface I between the two structures (which may be created by overlapping the material forming the flexible portion 12 of the bag on an inside or outside surface of the flange 22 to form an overlapping joint, or possibly in some cases by forming a butt joint).
- the connection may be made using well-known techniques, such as ultrasonic or thermal welding (heat or laser) at the interface to form a seal (which is at least liquid-impervious and preferably hermetic).
- the interface I may be used at the interface I, although this is obviously less preferred in view of the desirability in most cases for the more reliable, leak-proof seal afforded using welding techniques.
- the judicious use of inert sealants may be made along the joint thus formed to ensure that a leak-proof, hermetic seal results.
- the need for such an interface may be altogether eliminated by simply affixing the rigid portion 14 to an inside or outside surface of the bag 10 (see Figures 16a and 16b).
- the bag 10 shown in Figure 1 maybe manufactured as described above, with the fluid-agitating element 18 received on the post 20 (which may be accomplished using the techniques shown in Figures 1 b and 1 c).
- the empty bag 10 may then be sealed and folded for shipping, with the fluid-agitating element 18 held at the home location by the post 20. Holding in the axial direction (i.e., the vertical direction in Figure 1 ) may be accomplished by folding the bag 10 over the post 20, or by providing the portion 20a that is oversized or very close in size to the opening 18a in the fluid-agitating element 18.
- the bag 10 is then unfolded. It may then be placed in a rigid or semirigid support structure, such as a container C, partially open along at least one end such that at least the rigid portion 14 remains exposed (see Figure 2).
- Fluid F may then be introduced into the bag 10, such as through an opening or fitting (which may be a sterile or aseptic fitting, in the case where the bag 10 is pre-sterilized or otherwise used in a sterile environment).
- an opening or fitting which may be a sterile or aseptic fitting, in the case where the bag 10 is pre-sterilized or otherwise used in a sterile environment.
- An external motive device 24 is then used to cause the fluid-agitating element 18 (which is at least partially magnetic or ferromagnetic) to at least rotate to agitate any fluid F in the bag 10.
- the fluid-agitating element 18 is at least partially magnetic and is shown as being levitated by the motive device 24, which is optional but desirable.
- the levitation may be provided by a field-cooled, thermally isolated superconducting element SE (shown in phantom in Figure 2) positioned within the motive device 24 and thermally linked to a cooling source (not shown).
- the fluid-agitating element 18 may then be rotated by rotating the superconducting element SE (in which case the fluid-agitating element 18 should produce an asymmetric magnetic field, such as by using at least two spaced magnets having alternating polarities).
- a separate drive structure e.g., an electromagnetic coil
- a coupling capable of transmitting torque to the particular fluid-agitating element (which may be "levitated" by a hydrodynamic bearing; see, e.g., U.S. Patent No. 5,141 ,327 to Shiobara).
- a drive structure such as a shaft
- the particular means used to levitate and/or rotate the fluid-agitating element 18 is not considered critical to practicing the inventions disclosed herein.
- the fluid-agitating element 18 is also depicted as including a plurality of vanes or blades B to improve the degree of fluid agitation.
- the vanes or blades B preferably project in a direction opposite the corresponding surface of the rigid portion 14.
- the particular number, type, and form of the vanes or blades B is not considered important, as long as the desired degree of fluid agitation for the particular application is provided. Indeed, in applications where only gentle agitation is required, such as to prevent damage to delicate suspensions or to merely prevent stagnation of the fluid F in the bag 10, the vanes or blades B need not be provided, as a rotating smooth- walled annular element 18 still provides some degree of agitation. As explained above, it may be desirable to not only know the general location or position of the fluid-agitating element 18 within the bag 10, but also to assure its position relative to the motive device 24.
- the rigid portion 14 may be provided with a second receiver 26 to facilitate the correct positioning of the motive device 24 relative to the fluid-agitating element 18 when held at the home location.
- the second receiver 26 takes the form of a second post 28 projecting in a direction opposite the first post 20.
- the second post 28 is essentially coaxial with the first post 20 (although the post 20 maybe a separate component that fits into a receiver 14a defined by the second post 28; see Figure 1 c) and is adapted to receive an opening 24a, such as a bore, in the adjacent end face 24b forming a part of the housing for the motive device 24.
- the second post 28 helps to assure that the alignment between the fluid-agitating element 18 (which is generally held in the vicinity of the first receiver 16/post 20, which is the home location) and the motive device 14 is proper such that the desired coupling for transmitting the levitation or rotational force may be formed.
- the second receiver 26, such as second post 28 has a cross-sectional shape corresponding to the shape of the opening 24a.
- the second post 28 may be square in cross-section for fitting in a correspondingly- shaped opening 24a or locator bore.
- the second post 28 could have a triangular cross-sectional shape, in which case the opening 24a would be triangular.
- the second receiver 26 may be provided with a certain shape that corresponds only to the opening 24 in the motive device 24 having that type of superconducting element or drive structure.
- a similar result could also be achieved using the relative sizes of the second receiver 26 and the opening 24a, as well as by making the size of the opening 18a such that it only fits on a first receiver 16 having a smaller width or diameter, and then making the second receiver 26 correspond to an opening 24a in a motive device 24 corresponding to that element 18.
- an external structure is provided to which a motive device could be directly or indirectly attached and held in a suspended fashion (see, e.g., U.S. Patent No.4,209,259 to Rains et al., the disclosure of which is incorporated herein by reference). This structure serves to automatically align the motive device with the fluid-agitating element supported therein.
- a bag 10 per se is generally incapable of providing reliable support for the motive device 24, which can weigh as much as twenty kilograms.
- the motive device 24 in the embodiments disclosed herein for use with a vessel in the form of a bag 10 is generally supported from a stable support structure (not shown), such as the floor, a wheeled, height adjustable platform, or the like. Since there is thus no direct attachment with the bag 10, the function performed by the second receiver 26 in aligning this device with the fluid-agitating element 18 is an important one.
- FIGs 3a and 3b Another embodiment of the vessel forming one aspect of the present invention is shown in Figures 3a and 3b. In this embodiment, the vessel is again a bag 10 including a flexible portion 12 and a rigid portion 14.
- the rigid portion 14 is cap or hat-shaped with a peripheral flange 22 for attachment to the flexible portion 12 of the bag 10.
- the connection between the two structures may be formed using the various techniques described above, and preferably results in a fluid-impervious, hermetic seal.
- the rigid portion 14 includes a first receiver 16 in the form of a recess or cavity 30 facing the interior of the bag (see action arrow B) for receiving a correspondingly-shaped portion of the fluid-agitating element 18 in the bag 10 and holding it at a home location, at least when oriented as shown in Figure 3a.
- the portion of the fluid-agitating element 18 received in the cavity 30 is preferably the body 18b, which as described above is at least partially magnetic or ferromagnetic and may optionally support a plurality of vanes or blades B.
- the body 18b of the fluid- agitating element 18 is circular in cross-section and the cavity 30 is sized and shaped such that the body (which need not include opening 18a in view of the absence of post 20) may freely be inserted, rotate, and levitate therein.
- the fluid-agitating element 18 could also be in the form of a conventional magnetic stirrer (which of course would not be levitated), such as a bar having a major dimension less than the corresponding dimension (e.g., the diameter) of the cavity 30.
- the fluid-agitating element 18 in this embodiment is again free of direct attachment from the first receiver 16, but is held at a home location, even in the event decoupling.
- the fluid-agitating element 18 may be positioned in the first receiver 16 in the bag 10. The bag 10 may then be sealed, folded for storage or shipping, stored or shipped, and ultimately unfolded for use.
- the folding is preferably completed such that the fluid-agitating element 18 is captured in the cavity 30 and remains held in place during shipping by an adjacent portion of the bag 10. Consequently, upon unfolding the bag 10, the fluid-agitating element 18 is at the expected or home location, but remains free of direct attachment and ready to be rotated (and possibly levitated). If levitated, the levitation height established by the superconducting bearing or hydrodynamic bearing is preferably such that at least a portion of the body 18b of the fluid-agitating element 18 remains within the confines of the cavity 30. This helps to assure that the fluid-agitating element 18 remains held at the home location (that is, in the vicinity of the first receiver 16), even in the case of accidental decoupling from the motive device 24.
- the fluid-agitating element 18 will engage the sidewall of the cavity 30 and simply come to rest therein, which defines the home location. This not only improves the chance of an automatic recoupling, but also makes the task of manually reforming the coupling an easy one.
- An option to assure that a magnetic fluid-agitating element 18 remains associated with the first receiver 16, even if inverted, is to attach an attractive structure, such as a magnet 32 (shown in phantom in Figure 3 a), to the exterior of the rigid portion 14. The non-contact coupling thus established helps ensure that the fluid-agitating element 18 remains in the home location prior to being coupled to an external motive device.
- the magnet 32 is removed once the bag 10 is positioned on or in a support structure, such as a container C (see Figure 2). Such a magnet 32 may also be used with the embodiment of Figure 1 , which eliminates the need for providing the post 20 with portion 20a.
- the magnet 32 is preferably annular with an opening that is received by the second receiver 26, which advantageously helps to ensure the proper alignment for forming the coupling.
- Yet another option is to provide a frangible adhesive on the fluid-agitating element 18 to hold it in place temporarily in the first receiver 16 prior to use. The strength of any adhesive used is preferably such that the bond is easily broken when the fluid-agitating element 18 is levitated in the first receiver 16.
- the first receiver 16 in this embodiment also serves the dual function of helping to align the fluid-agitating element 18 relative to an external motive device 24.
- the periphery of the sidewall 34 and the end wall 36 defining the cavity 30 in the rigid portion 14 define a second receiver 26 adapted to receive an opening 24a formed in an adjacent face of a motive device 24.
- the opening 24a is preferably sized and shaped for being received by the second receiver 26, and may even help to ensure that the bag 10 is used only with a motive device 24 having the correct superconducting element or magnetic structure(s) for levitating and/or rotating the fluid-agitating element 18.
- the opening 24a is also cylindrical.
- the opening 24a also has a depth such that the end wall 36 rests on the corresponding face 24c of the motive device 24. This feature may be important to ensure that the gap between the superconducting element and/or drive structure in the motive device 24 and the at least partially magnetic or ferromagnetic body 18b of the fluid-agitating element 18 is minimized, which helps to ensure that the strongest possible coupling is established and that the maximum amount of driving torque is transferred.
- the gaps are shown as being oversized in Figure 3b merely to provide a clear depiction of the relative interaction of the structures shown.
- Figures 4a and 4b show an embodiment similar in some respects to the one shown in Figure 3 a and 3b.
- the rigid portion 14 includes a peripheral flange 22 connected to the flexible portion 12 of the bag 10 to form a seal.
- the rigid portion 14 includes a sidewall 34 and end wall 26 that together define a cavity 30.
- a major difference is that the cavity 30 of the rigid portion 14 essentially faces outwardly, or toward the exterior of the bag 10 (e.g., in a direction opposite action arrow B).
- the sidewall 34 and end wall 36 define the first receiver 16 for receiving the fluid-agitating element 18, which is shown having an annular body 18b that is at least partially magnetic or ferromagnetic and may support a plurality of vanes or blades B.
- the first receiver 16 in the form of the periphery of the sidewall 34 provides a similar receiving function as both the post 20 and the cavity 30 of the other embodiments, since it is capable of maintaining, holding, or confining the fluid-agitating element 18 substantially in a home or expected position within the bag 10.
- the maximum amount of side-to-side movement is of course dependent on the size of the opening 18a in the fluid-agitating element.
- the outwardly- facing cavity 30 is adapted to serve as the second receiver 26 for receiving a portion of a motive device 24 used to levitate and rotate the fluid-agitating element 18 and serving to align the two.
- the motive device 24 may include a head end 24d adapted for insertion in the cavity 30 to form the desired coupling with the fluid-agitating element 18 positioned adjacent thereto.
- the spacing between the head end 24d and at least the sidewall 34 is preferably minimized to maximize the strength of the coupling between the motive device 24 and the fluid-agitating element 18.
- the end face 24b of the head end 24d may rest against and assist in supporting the bag 10 (which, as described above, may be positioned in a separate, semi-rigid container (not shown)).
- the possible use of a levitating fluid-agitating element 18 with a superconducting bearing or a hydrodynamic bearing is described. In such systems, a real possibility exists that the fluid-agitating element 18 might accidentally decouple or disconnect from the motive device 24, such as if the fluid is viscous or the amount of torque transmitted exceeds the strength of the coupling.
- an added advantage of the use of the first receiver 16 in each of the above-referenced embodiments is that, despite being free from direct attachment, it still serves the function of holding the fluid-agitating element 18 at the home location in instances where accidental decoupling occurs. This significantly reduces the downtime associated with such an event, since the general position of the fluid-agitating element 18 is known.
- first receiver in the bag 10 also improves the chances of automatic recoupling, since the fluid-agitating element 18 remains generally centered relative to the motive device 14 and held generally at the home location, even when decoupling occurs.
- a related advantage is provided by forming the first receiver 16 in or on a rigid portion 14 of the bag 10. Specifically, in the case where a fluid-agitating element rests on a surface of a bag, the contact over time could result in damage and could even lead to an accidental perforation, which is deleterious for obvious reasons. The possibility for such damage or perforation also exists when a levitating fluid-agitating element 18 accidentally decouples.
- the potential for such damage or perforation is substantially eliminated in the foregoing embodiments, since the first receiver 16 helps to keep the fluid-agitating element 18 adjacent to the flange 22 of the rigid portion 14, which is generally thicker and less susceptible to being damaged or perforated. In other words, if the fluid-agitating element 18 becomes decoupled, it only engages or contacts the rigid portion 14 of the bag 10. Thus, it is preferable for the flange 22 to be oversized relative to the fluid-agitating element 18. While the embodiments of Figures 1-4 are described as bags 10 including both a flexible portion 12 and a rigid portion 14, it should be appreciated that the present invention extends to a completely rigid vessel (that is, one made of metal, glass, rigid plastics, or the like).
- the post 20 preferably includes a portion 20a for capturing the fluid-agitating element 18 thereon, but without any other means of direct attachment or bearing.
- the focus has been on a fluid-agitating element 18 capable of levitating in the vessel.
- the inventions described herein may also be applied to a bag 10 in combination with a fluid-agitating element 18 directly supported by one or more bearings.
- the first receiver 16 associated with the rigid portion 14 of the bag 10 may be in the form of an inwardly-projecting post 20 including a slide bearing 40 for providing direct support for the fluid-agitating element 18.
- the bearing 40 is preferably sized and shaped such that it fits into an opening 18a forming in the fluid-agitating element 18, which may rest on the adjacent surface of the post 20 or may be elevated slightly above it. In either case, it should be appreciated that the first receiver 16 receives and holds the fluid- agitating element 18 in a home location, both during shipping and later use.
- the material forming the slide bearing 40 is preferably highly wear-resistant with good tribological characteristics. The use of a slide bearing 40 is preferred in applications where the bag 10 is disposable and is merely discarded, since it is less expensive than a corresponding type of mechanical roller bearing (and is actually preferred even in the case where the bag 10 is reused, since it is easier to clean).
- the rigid portion 14 of the bag 10 in this embodiment may further include a second receiver 26 in the form of a second post 28 coextensive and coaxial with the first post 20.
- the second post 28 is received in an opening 24a formed in an end face 24b of a motive device 24.
- the motive device 24 in this case includes only a drive structure DS (shown in phantom in Figure 5b) for forming a coupling with the body 18b, which is magnetic or ferromagnetic (iron, magnetic steel, etc.).
- the drive structure DS may be a permanent magnet or may be ferromagnetic, as necessary for forming the coupling with the fluid-agitating element 18, which may be disc-shaped, cross-shaped, an elongated bar, or have any other suitable shape.
- the drive structure DS may be rotated by a direct connection with a motor (not shown), such as a variable speed electric motor, to induce rotation in the fluid-agitating element 18.
- the drive structure DS may be an electromagnet with windings to which current is supplied to cause the magnetic fluid-agitating element 18 rotate and possibly levitate slightly to create a hydrodynamic bearing (see, e.g., U.S. Patent No. 5,141,327, the disclosure of which is incorporated herein by reference).
- the particular type of motive device 24 employed is not considered critical to the present invention.
- Figures 6a and 6b show an embodiment of the bag 10 in which the first receiver 16 is in the form of a cavity 30 formed in the rigid portion 14 and facing inwardly.
- a bearing 40 is provided in the cavity 30 for providing direct support for a fluid-agitating element 18 positioned therein.
- the bearing 40 may be a slide bearing adapted for insertion in the opening 18a of the fluid-agitating element 18 formed on the head end of a post 42.
- the post 42 may be supported by or unitarily formed with the end wall 36.
- the periphery of the sidewall 34 also defines a second receiver 26 for receiving a corresponding opening 24a in a motive device 24, which in view of the direct support provided by bearing 40 need only provide the force necessary to rotate the fluid-agitating element 18 in a non-contact fashion.
- the embodiment shown in Figures 7a and 7b is the direct support counterpart for the embodiment shown in Figures 4a and 4b.
- the rigid portion 14 again includes a cavity 30 facing outwardly or toward the exterior of the bag 10 and a first receiver 16 for receiving and defining a home location for a fluid-agitating element 18.
- the first receiver 16 includes a bearing 40 for supporting the fluid-agitating element 18, which again is at least partially magnetic or ferromagnetic.
- the bearing 40 may be a slide bearing formed on the head end of a post 44 integral with the end wall 36 of the rigid portion 14 and adapted for fitting into an opening or recess 18a in the fluid-agitating element 18, or may be a different type of bearing for providing support therefor.
- the motive device 24 includes a head end 24d adapted for insertion in a second receiver 26 defined by the cavity 30.
- This head end 24d preferably includes the drive structure DS that provides the force for causing the at least partially magnetic or ferromagnetic fluid-agitating element 18 to rotate about bearing 40.
- the fluid-agitating element 18 includes an optional depending portionl 8b that extends over the sidewall 34. As should be appreciated, this portion may also be magnetized or ferromagnetic such that a coupling is formed with the drive structure DS.
- a similar type of fluid-agitating element 18 could also be used in the levitation scheme of Figures 4a and 4b.
- the sparger 100 includes a face portion 102a for attaching to the bag 10 and a tubular projecting portion 102b or coupling with an external source of gas, such as through a tube U.
- the face portion 102a may comprise a disk-shaped piece of rigid plastic material, and may be welded directly to the flexible material forming the bag 10 adjacent the fluid F when present such that a fluid-impervious seal results.
- Gas introduced through the tube U from a remote source (not shown) thus enters the bag 10, passing through any fluid present.
- the means for forming bubbles in the gas entering the fluid comprises a perforated piece of plastic film 104 may also be secured adjacent the face portion 102a of the sparger 100, such as by welding.
- the holes in the film 204 are preferably in the sub- millimeter range.
- a porous film may be used, various types of which are generally well known in the art (see, e.g., U.S. Patent No. 4,814,124, incorporated herein by reference).
- the bag 10 with the integral sparger 100 may simply be pre-sterilized with the impeller 18, and then disposed upon recovering all or part of the fluid, or alternatively a product therefrom once mixing is complete.
- the rigid portion 14 is welded to the bag 10, as described above, preferably along the bottom and such that a fluid-impervious seal is formed.
- a receiver in the form of a post 20 may be removably attached to the rigid portion 14 through the opening (not shown) in the fluid-agitating element 18 (such as by providing a threaded bore in the rigid portion for receiving a threaded end of the post, or as shown in Figure lc, a bore 14a having a groove for establishing a snap-fit engagement with the post), and may include an oversized head for providing a retaining function.
- a second receiver 26 in the form of a recess or cavity in the rigid portion 14 may also be provided for receiving an external motive device, such as a rotating drive magnet or superconducting element (not shown).
- the face 14a of the rigid portion 14 further includes a first passage 14b in communication with both the interior of the bag 18 and a second passage 14c leading to an external source of gas via a tube U. Gas introduced through the tube U thus exits into the interior of the bag 10 through the first passage 14b, which may be associated with a perforated piece of material 106 comprising the means for forming the bubbles.
- the first passage 14b is annular and includes seating ledges 14d for receiving the material 106, which takes the form of an annular piece of plastic film having a plurality of holes or apertures to form the perforations (which, again, are preferably sized in the sub-millimeter range) that is welded in place.
- the sparger 100 thus created in the illustrated embodiment releases the bubbles in close proximity to the fluid- agitating element 18, thus enhancing their dispersion throughout the fluid.
- the sparger 100 is integral with the rigid portion 14 connected to the bag 10, which may again include a receiver in the form of an inwardly projecting post 20 for receiving the fluid-agitating element 18 and an outwardly directed alignment structure 26.
- the post 20 in this embodiment includes an inlet 20a for connecting with a tube U coupled to an external source of gas (not shown), such as through an opening or open end of the bag, and an outer wall 20b formed of a perforated or gas permeable material that serves as the means for forming bubbles in this embodiment.
- a center support 20c may also be provided concentric with the outer wall 20b.
- gas passing through the tube U exits the outer wall 20b of the post 20 as bubbles (the size of which depend on the size of the perforations made, which again are preferably in the sub- millimeter range).
- the fluid-agitating element 18 when the fluid-agitating element 18 is levitated and/or rotated by an adjacent, but external motive device 24, the bubbles are released adjacent the opening 18a and dispersed throughout the fluid F.
- the bag 10 Once use of the bag 10 is complete, it may then simply be discarded along with the sparger 100.
- the bag includes a rigid receiver 200 creating a port for receiving a sensor S, such as one comprised of a translucent or transparent (preferably glass), close-ended tube G and a fiber optic cable L for transmitting light to and receiving back the reflected light (note bidirectional arrows).
- the receiver 200 includes a face 202a having a periphery to which the bag 10 is attached, such as by welding, to form a fluid impervious seal.
- a tubular portion 202b of the receiver 200 receives a bushing 204, which in turn receives the closed end of the transparent tube G and allows it to pass into contact with the fluid F when present in the bag 10.
- a sealing tube 206 couples the bushing 204 to the tubular portion 202b of the receiver 200 (which may include a slightly oversized, frusto-conical portion defining a hold-assist ledge for the tube).
- Fasteners such as cable ties 208, may help to removably secure the sealing tube 206 in place, although other means for sealing could be used instead, such as adhesives or the like.
- the important point is that no appreciable amount of fluid can pass the tube G once inserted in the receiver 200.
- the tube G may carry a fluorescent sensing element E, preferably by way of external attachment to the translucent or transparent closed end.
- this element E may change its fluorescence characteristics in response to change in the pH, dissolved oxygen, carbon dioxide, or temperature of the fluid it is touching. The fluorescence characteristics can then be measured by external apparatus capable of illuminating the sensitive element E, such as through the cable L, and the closed end of the tube G.
- this type of sensitive element E is not only disposable, but also advantageously does not require any power or leads.
- An example of an off-the-shelf sensor element E is one manufactured by PreSens (or Precision Sensing) GmbH Josef-Engert-Str.
- the rigid portion 14 as part of the bag 10 by forming a seal at an interface between the two, it could also be positioned in contact to an inner or outer surface of the bag and attached using vacuum-forming techniques, adhesives, or the like.
- the bag 10 would essentially line the inside surfaces of the sidewall 34 and end wall 36 (see Figure 16a).
- the bag 10 would cover the sidewall 34 and end wall 36 (see Figure 16b) . In both cases, the need for the flange 22 may be eliminated.
- any of the first receivers with a tapered or frusto-conical engagement surface that mates with a corresponding surface on the fluid-agitating element, as disclosed in my co-pending patent application Ser. No. PCT/US01/31459, the disclosure of which is incorporated herein by reference.
- the integral sparger 14 may also be provided in the embodiment in which the rigid portion 14 is cap or cup-shaped, such as by providing the perforated/permeable material (whether film, rigid, or otherwise) for forming the bubbles along the peripheral flange 22, sidewall 34, or end wall 36, and providing a passage to allow for gas to communicate with it from a remote source (such as through an external tube).
Abstract
Description
Claims
Priority Applications (11)
Application Number | Priority Date | Filing Date | Title |
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EP05705228.4A EP1701780B8 (en) | 2004-01-07 | 2005-01-07 | Bioprocessing vessel with integral sparger, and method of its manufacture |
JP2006549426A JP2007522801A (en) | 2004-01-07 | 2005-01-07 | Mixing bag with integral sparger and sensor receptacle |
CA2552717A CA2552717C (en) | 2004-01-07 | 2005-01-07 | Mixing bag with integral sparger and sensor receiver |
US11/304,417 US7469884B2 (en) | 2004-01-07 | 2005-12-15 | Mixing bag with integral sparger and sensor receiver |
IL176667A IL176667A (en) | 2004-01-07 | 2006-07-02 | Mixing bag with integral sparger and sensor receiver |
US11/746,898 US7384027B2 (en) | 2004-01-07 | 2007-05-10 | Mixing bag with integral sparger and sensor receiver |
US12/341,478 US7992846B2 (en) | 2004-01-07 | 2008-12-22 | Mixing bag with integral sparger and sensor receiver |
US12/761,111 US8123199B2 (en) | 2004-01-07 | 2010-04-15 | Bioreactor |
US13/405,609 US8550439B2 (en) | 2004-01-07 | 2012-02-27 | Mixing bag with integral sparger and sensor receiver |
US13/599,652 US8708319B2 (en) | 2004-01-07 | 2012-08-30 | Mixing bag with integral sparger and sensor receiver |
US14/263,499 US9770696B2 (en) | 2004-01-07 | 2014-04-28 | Mixing bag with integral sparger and sensor receiver |
Applications Claiming Priority (4)
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US53503104P | 2004-01-07 | 2004-01-07 | |
US60/535,031 | 2004-01-07 | ||
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US60/599,960 | 2004-08-09 |
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US11/304,417 Continuation US7469884B2 (en) | 2004-01-07 | 2005-12-15 | Mixing bag with integral sparger and sensor receiver |
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WO2005068059A1 true WO2005068059A1 (en) | 2005-07-28 |
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PCT/US2005/000464 WO2005068059A1 (en) | 2004-01-07 | 2005-01-07 | Mixing bag with integral sparger and sensor receiver |
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US (7) | US7469884B2 (en) |
EP (2) | EP1701780B8 (en) |
JP (1) | JP2007522801A (en) |
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IL (1) | IL176667A (en) |
WO (1) | WO2005068059A1 (en) |
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CA2552717A1 (en) | 2005-07-28 |
EP2813281B1 (en) | 2016-08-17 |
EP1701780B8 (en) | 2014-09-24 |
IL176667A0 (en) | 2006-10-31 |
US7469884B2 (en) | 2008-12-30 |
IL176667A (en) | 2012-03-29 |
US20100197003A1 (en) | 2010-08-05 |
US20130171723A1 (en) | 2013-07-04 |
US20060131765A1 (en) | 2006-06-22 |
US7384027B2 (en) | 2008-06-10 |
JP2007522801A (en) | 2007-08-16 |
US20090135667A1 (en) | 2009-05-28 |
US7992846B2 (en) | 2011-08-09 |
US8550439B2 (en) | 2013-10-08 |
US20120152769A1 (en) | 2012-06-21 |
EP1701780A4 (en) | 2011-11-23 |
US8123199B2 (en) | 2012-02-28 |
CA2552717C (en) | 2011-11-29 |
EP1701780B1 (en) | 2014-07-30 |
US20070252290A1 (en) | 2007-11-01 |
US20140234951A1 (en) | 2014-08-21 |
US8708319B2 (en) | 2014-04-29 |
EP2813281A1 (en) | 2014-12-17 |
EP1701780A1 (en) | 2006-09-20 |
US9770696B2 (en) | 2017-09-26 |
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