WO2013165310A1

WO2013165310A1 - Container comprising a tubing arrangement

Info

Publication number: WO2013165310A1
Application number: PCT/SE2013/050484
Authority: WO
Inventors: Colin R. Tuohey; Thomas Erdenberger; Richard L. Damren; Parrish M. Galliher
Original assignee: GE Healthcare Bio Sciences Corp; Xcellerex Inc
Current assignee: Global Life Sciences Solutions USA LLC
Priority date: 2012-05-04
Filing date: 2013-05-02
Publication date: 2013-11-07
Anticipated expiration: 2014-11-04

Description

CONTAINER COMPRISING A TUBING ARRANGEMENT

TECHNICAL AREA The present invention relates to a container comprising a tubing arrangement, and a mixing vessel and/or a bioreactor comprising the container.

BACKGROUND A variety of vessels, devices, components and unit operations for manipulating fluids and/or for carrying out chemical, biochemical and/or biological processes are available. For instance, biological materials (e.g., animal and plant cells) including, for example, mammalian, plant or insect cells and microbial cultures can be processed using bioreactors. Traditional bioreactors, which are typically designed as stationary reusable tanks or containers, or disposable bioreactors, many of which utilize plastic sterile bags or containers, may be used for such purposes. Although chemical manufacturing systems, pharmaceutical manufacturing systems, bioreactor systems and other fluid manipulating systems (e.g., mixing systems) are known, improvements to such systems would be beneficial.

For such systems it is essential that they maintain sterile, aseptic, substantially particle- free around or in the system, especially for applications involving cell culture, clean manufacturing processes, and use of dangerous materials and/or infectious agents. During recent years, disposable or single use appliances in connection with the above mentioned systems have been used increasingly. In such systems it is essential that the performance of the system is kept stable while costs of the appliances are kept as low as possible. Therefore, there is a need to improve such systems and the

performance of such systems. Such systems comprising environmental containment systems are described in for example US2008/0139865 A1 , which is incorporated herein by reference. SUMMARY OF THE INVENTION

It is thus an object of the present invention to provide an improved system comprising a container for holding a fluid and useable in for example traditional bioreactors, which are designed as stationary reusable tanks or containers, or disposable bioreactors, which utilize plastic sterile flexible bags or containers. Especially, it is an object of the present invention to provide the container with improved sparging of a fluid, i.e. a liquid or a gas, into the container. It is a further object of the invention to improve a mixing operation inside the container comprising a mixer.

It is a further object of the invention to prevent entangling of tubings inside the container during shipping and use of the container.

The objectives above are at least mainly attained by the present invention that relates to a container comprising a tubing arrangement, the container capable of holding a fluid and comprising a container wall having an outer wall surface and an inner wall surface, the inner wall surface defining an interior chamber for holding the fluid. The container comprises at least two ports configured to allow a fluid to flow into the interior chamber, wherein the at least two ports are provided in the container wall. The arrangement also comprises at least two sparge tubes to provide the fluid in the interior chamber, and each of the sparge tubes, respectively, is connected to a port in the interior chamber. The arrangement further comprises at least one body member which couples the at least two sparge tubes to keep the sparge tubes in a fixed location. By this

arrangement it is possible to keep the sparge tubes tethered, i.e. in a fixed position, and thus for example entanglement of the tubes inside the container is prevented.

The tubing arrangement may further comprise at least one fastening element fixed to the container wall and arranged to be attached to the body member. The fastening element thus connects the body member to the container wall. The fastening element may have different heights or may have adjustable height and thus makes it possible to keep the body member and the sparge tubes connected to the body member at a desired distance from the container wall. The tubing arrangement may comprise at least two body members and each of the body members is configured to keep at least two sparge tubes in a fixed location. By providing several body members, the stability of the tubing arrangement may be improved.

The at least two body members are preferably connected to each other, e.g. by means of the sparge tubes, whereby the sparge tubes in the tubing arrangement are kept in a fixed location in an easy way. Also, this further improves the stability of the tubing arrangement.

The container may comprise a mixer comprising an axially extending mixer shaft and radially extending impeller blades. In one embodiment of the invention, the tubing arrangement may comprise at least four ports provided in the container wall, wherein the ports are connected to a respective sparge tube. Preferably, the ports surround the mixer radially or in a radial direction, whereby sparging can be provided in an even manner. In this way it is possible to improve the stability of the tubing arrangement, and thus further prevent entanglement of the tubes inside the container. Also entanglement with other appliances, such as a mixer, inside the container can be prevented.

The sparge tubes may be arranged underneath impeller blades in an axial direction of the mixer or a mixer shaft of the mixer, or sparge tubes may be located above the impeller blades in the axial direction of the mixer. Preferably, the gas is directed towards the center of the impeller.

The body member may comprise a through hole that extends perpendicular to the lengthwise direction of the body member. This makes it possible to couple tubes or shafts to the body member and thus obtain a desired structure for the tubing

arrangement. This also enables e.g. the connection of tubes in an easy way. The through hole is preferably configured to allow the fluid to pass through the hole, i.e. so that a sparge tube can be arranged to pass through the hole. Thus, this construction of the body member does not prevent the fluid from passing the body member and the body member may be used to connect tubes and/or body members in a space saving manner while the stability of the arrangement can be further improved.

The sparge tube may comprise a tee fitting to direct the fluid to a desired direction. In this way, it is possible to improve the sparging of the fluid inside the container, since the direction of the flow of the fluid may be easily adjusted. Alternatively or additionally the sparge tube may comprise at least one and preferably a plurality of orifices to direct the fluid to a desired direction. Thus, sparging of fluid can be obtained towards the desired direction. The orifices may be drilled to the sparge tube and the sparging can be adapted to the desired process in an easy and cost saving way. The structure of the tubing arrangement can thus also be simplified.

The sparge tube preferably comprises an end cap. Through this arrangement it is possible to prevent fluid from escaping in the wrong location and prevent liquid build-up in the sparge tube.

The body member may comprise at least one hose barb fitting protruding outwards from the body member for attaching the body member to a further tube.

The port in the container wall may comprise a hose barb fitting for attaching the sparge tube to the port. In this way a leak tight attachment can be provided in an easy and low- cost manner.

The at least one body member is preferably a rigid body member. In this way, the stability of the arrangement can be improved.

The container is preferably flexible. In this way the container may be disposable and it may be shipped to a customer in a space saving manner.

The present invention also relates to a mixing vessel and to a bioreactor comprising the container as described above.

Further advantages and objects of the present invention are described in the following detailed description of the invention with reference to the attached drawings. BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 is a schematic illustration of a bioreactor with an environmental containment system in which the container according to the present invention is useable.

Fig. 2 is a schematic perspective illustration of a mixing vessel comprising a container with a tubing arrangement according to the present invention.

Fig. 3 illustrates schematically a partially cross-sectioned top view of a tubing

arrangement in a container according to the present invention.

Fig. 4 illustrates schematically a partially cross-sectioned, enlarged and cut side view of a fastening means according to the present invention. Fig. 5A illustrates a side view of the body member according to the present invention.

Fig. 5B illustrates a top view of the body member shown in Fig. 5A.

DETAILED DESCRIPTION

A container comprising a tubing arrangement, which is herein also referred to a tethered sparge tube arrangement, is suitable for use for example in an environmental containment system shown in Fig. 1 . These systems may be self-sufficient and independently customized to perform a specific biological, chemical, or pharmaceutical process. Such systems may be customized to perform a particular process at a first location, disassembled, and then shipped to a second location to perform the same process at the second location. Furthermore, the use of apparatuses having an environmental containment enclosure can allow the apparatuses to be used in non- sterile or non-clean room environments for processes requiring such environments, since the enclosed space(s) formed by the environmental containment enclosure(s) can be operated under sterile, aseptic, particle-free, or reduced-particle conditions. This feature can substantially save costs as clean room or other facilities may not be required. The container of the present invention can also be used in traditional clean room arrangements, with or without a supporting vessel. The container according to the present invention may be used in vessels which are a part of a bioreactor system. Bioreactors may be used to produce a variety of products from various organisms such as, for example, bacteria, insects/insect cells, fungi, mammalian cells, human cells, yeast, shrimp, fish, protozoa, nematodes, viruses, algae, and higher plants/plant cells. The container may also be used in other types of systems e.g. in those involving containment systems in general, systems for

containment and/or processing of a fluid (e.g., a liquid or a gas) in mixing systems, systems related to a biological, chemical, and/or pharmaceutical manufacturing process, e.g., primary recovery, filtration and chromatography systems, cell culture systems, microscopy and/or other analytical devices, etc., as well as other applications that require sterile, aseptic, substantially particle-free, and/or reduced-particle environments. It should also be understood that while many examples provided herein involve the use of vessels comprising collapsible bags or flexible containers, aspects of the invention can be integrated with vessels comprising non-collapsible or rigid containers, and other configurations involving liquid containment. Fig. 1 shows a non-limiting example of an inventive bioreactor system including a container, such as a flexible container, and an environmental containment enclosure. The apparatus 100 includes a first containment apparatus in the form of a vessel 1 14, which, in the illustrated embodiment, is a reusable support structure (e.g., a stainless steel tank) that surrounds and contains a container 1 18. Apparatus 100 also includes a second containment apparatus in the form of an environmental containment enclosure 120, which surrounds a portion of vessel 1 14.

The container 1 18 is configured as a collapsible bag (e.g., a polymeric bag).

Additionally or alternatively, all or portions of the collapsible bag or other container may be formed of a substantially rigid material such as a rigid polymer, metal, and/or glass. Container 1 18 may be disposable and may be configured to be easily removable from support structure 1 14. If a collapsible bag is used, collapsible bag 1 18 is fluid tight to enable it to contain a liquid 122, which may contain reactants, media, and/or other components necessary for carrying out a desired process such as a chemical, biochemical and/or biological reaction. Collapsible bag 1 18 may also be configured such that liquid 122 remains substantially in contact only with the collapsible bag during use and is not in contact with support vessel 1 14. The bag may be disposable and used for a single reaction or a single series of reactions, after which the bag is discarded. Because the liquid in the collapsible bag does not come into contact with the support structure 1 14, the support structure can be reused without cleaning. That is, after a reaction takes place in the container 1 18, the container can be removed from support structure 1 14 and replaced by a second (e.g., disposable) container. A second reaction can be carried out in the second container without having to clean either the first container or the reusable support structure. Also shown in FIG. 1 are an optional inlet port 142 and optional outlet port 146, which can be formed in container 1 18 and/or reusable support structure 1 14 and can facilitate more convenient introduction and removal of a liquid and/or gas from the container. The container may have any suitable number of inlet ports and any suitable number of outlet ports. For example, one or more connections 164 may be positioned at a top portion of container 1 18 or at any other suitable location. Connections 164 may include openings, tubes, and/or valves for adding or withdrawing liquids, gases, and the like from container 1 18, each of which may optionally include a flow sensor and/or filter (not shown). Optionally, connections 164 may be in fluid communication with gas

introduction and withdrawal ports 165.

In accordance with the present invention a plurality of inlet ports may be used to provide different gas compositions via a plurality of spargers 147, which will be described in more detail below. The spargers 147 allow separation of gases prior to their introduction into the container. These spargers 147 may be positioned at any suitable location with respect to container 1 18. Preferably, the container includes two or more spargers 147 located at a bottom portion, i.e. in the lower half of the container height, of the container. The spargers 147 may be associated with sparge tubes which may be connected to the container wall via ports, for example, the sparge tubes may be connected to inlet and/or outlet ports to form, e.g., delivery and harvest lines, respectively, for introducing and removing liquid from the container.

The support structure 1 14 and/or container 1 18 may also include one or more ports 154 that can be used for sampling, analyzing (e.g., determining pH and/or amount of dissolved gases in the liquid), or for other purposes. These ports may be aligned with one or more access ports 156 of environmental containment enclosure 120.

Optionally, the container and/or support structure may include a utility tower 150, which may be provided to facilitate interconnection of one or more devices internal to the container and/or support structure with one or more pumps, controllers, and/or electronics (e.g., sensor electronics, electronic interfaces, and pressurized gas controllers) or other devices. Such devices may be controlled using a control system 134.

The container 1 18 according to the present invention may be used in systems including multiple sparges 147. A control system 134 may be operatively associated with each of the sparges 147 and configured to operate the sparges 147 independently of each other. This can allow, for example, control of multiple gases being introduced into the container.

Apparatus 100 may optionally include a mixing system such as an impeller 151 positioned within container 1 18, which can be rotated (e.g., about a single axis) using a motor 152 that may be external to the container 1 18. In some embodiments, as described in more detail below, the impeller and motor are magnetically coupled. The mixing system can be controlled by control system 134.

Fig. 2 illustrates the container 1 18 according to one embodiment of the invention in more detail. The container 1 18 comprises a container wall 218 that extends over the whole container 1 18. The container 1 18 is flexible and collapsible and comprises a tubing arrangement 200 with two sparge tubes 220 and two rigid body members 210 that are coupled together by means of the sparge tubes 220 so that the sparge tubes 220 are kept in fixed location. By rigid is meant a material that is not bending, or a material that is stiff. The sparge tubes 220 are coupled to ports 230 that penetrate the container wall and thus allow a fluid to enter the container. The ports 230 are connected to the sparge tubes 220. The ports are connected to a fluid source, such as oxygen gas, so that fluid can be provided into the container. The ports 230 couple the sparge tubes 220 in a fluid tight manner, e.g. by means of hose barb fittings, so that the interior chamber of the container and thus the process can be kept sterile.

In the container shown in Fig. 2, the tubing arrangement 200 surrounds a mixer 151 in a radial direction. The mixer includes a mixer shaft 157 and an impeller 155 with impeller blades. As shown in the drawing, the sparge tubes 220 are connected to each other via two body members 210 and two tube legs, i.e. sparge tubes 224, that extend in a perpendicular direction in respect to the longitudinal direction of the tubes 220, i.e. in the same direction as the body members 210. Thus a tee coupling is formed to couple the tubes 220 and 224 to each other. In the embodiment shown in Fig. 2, the sparge tubes 224 are drilled in proximity to the tee to provide orifices to the tubes. The orifices are drilled to the tubes 224 so that fluid or gas may be released towards the center of the impeller 155, and thus the orifices in the tubes form a sparger 147 that releases fluid to the container.

In another embodiment, the tubes may be provided with a tee fitting to provide a sparger 147. This embodiment will be described in more detail with reference to Fig. 3 below.

Further shown in Fig. 2 are end caps 222 that cap the sparge tubes 220 and prevent the gas from escaping in the wrong direction and prevent liquid build-up in the sparge tubes 220. Also the arrangement comprises four fastening elements 232, such as hose barbs, fixed to the container wall 218 by means of welding. The fastening elements 232 are connected to the body member 210 by means of tubes or other type of connectors. The connectors may be capped on the opposite side of the body member 210 with a cap 234. The fastening element thus indirectly connects the body member 210 to the container wall 218. The fastening element 232 may have a desired height or may have adjustable height, e.g. by means of a telescopic construction, and thus makes it possible to keep the body member 210 and the sparge tubes 220 connected to the body member at a desired distance from the container wall 218. The container further comprises an outlet 146 supported by a supporting means 148. The construction of an alternative tubing arrangement 200 useable in the arrangement shown in Fig. 2 is shown in more detail in Fig. 3 which shows the embodiment in a view from above, whereby the container wall 218 corresponds to the side wall of the container as shown in Fig. 2.

The spargers 147 may be obtained as follows. The container wall 218 of the container 1 18 may be penetrated by means of a welded hose barb port 230 with ports on either side of the container wall 218. Generally, another way of penetrating the container wall 218 could be by providing a bulk head style non-integral fitting with ports on either side of the container wall 218. In the embodiment shown in Fig. 2, the container 1 18, or the container wall 218, comprises two ports, wherein each of the ports 230 is configured to allow a fluid to flow into the interior chamber of the container 1 18. As further shown schematically in Fig. 3, the arrangement 200 comprises two sparge tubes 220 that are connected to the respective port 230 in the interior chamber of the container 1 18 by means of a hose barb fitting. The sparge tubes 220 provide fluid in the interior chamber of the container 1 18. The arrangement further comprises two body members 210 which couple the sparge tubes 220 and keep the sparge tubes 220 in a fixed location.

The arrangement 200 may comprise a plurality of body members, whereby the stability of the tubing arrangement can be improved. Also, the ports 230 could be placed at the bottom of the container 1 18 and the tubing arrangement 200 could be constructed such that it extends vertically (or in axial direction) in a multiple level construction, e.g. over the part or over the entire height of the container. Thus, tethered sparge tubes 220 are provided and a means for gas to be injected into the disposable bioreactor or fermentor from any boundary surface of the bag is provided. As further shown in Fig. 3, internal to the container 1 18, the sparge tube 220 comprises two legs 220a, 220b with a tee fitting 240 in the middle. The tee fitting 240 in the middle makes it possible to direct the gas or fluid in the tube towards the center of the arrangement 200 in an easy way. In this arrangement, no tubes or legs 224 are needed, but could be included if desirable. Other constructions could be used to enable an easy direction of the gases, and for example a valve construction could be used, e.g. one involving a three way valve. This could be beneficial for example in the arrangement shown in Fig. 2, in which the arrangement 200 comprises an impeller shaft 157 in the center of the tubing

arrangement 200.

Generally, when gases (or fluids) are directed towards the center of the mixer or impeller 155 the mixing of the gases in the container can be improved. Of course, the gas could be directed towards any other advantageous location for optimal gas distribution in the liquid phase. Also, the sparge tube 220 may comprise several legs and tee fittings as well as body members 210, whereby a construction of variable height can be provided inside the container while the construction remains stable. The end of each sparge tube 220 may be capped by means of a cap 222 to prevent gas from escaping from the tube in a wrong location and to prevent liquid build-up in the sparge tube 220.

To further improve the stability of the tubing arrangement 200 shown in Fig. 3, the sparge tubes 220 may be fixed to the inner wall of the container 1 18 by means of fastening means 232, as illustrated in Fig. 4. The body members 210 secure the sparge tubes 220 in place.

In Fig. 4 the construction for the fastening means 232 for the connection between the body member 210 and the container wall 218 is illustrated schematically.

It should be noted that the dimensions of the sparge tubes and tubes in general are enlarged in all the drawings. It should be noted that in fact the connection between the hose barbs and the tubes is liquid tight and thus the wall of the tube and the hose barb are in connected without passages between the wall of the tube and the hose barbs.

Fig. 4 shows the arrangement partially cut as indicated by the line on the right hand side of the figure. As can be seen, the container 1 18 or the container wall comprises hose barb 252 and a tube 254, and the tube 252 is fixedly attached to the hose barb 252. The body member 210 comprises a hose barb fitting 250 to enable coupling to the tube 254, and is also fixedly attached to the tube 254. The body member comprises a through hole 260 that extends perpendicular to the lengthwise direction of the body member 210. The lengthwise direction is illustrated in Fig. 3 and 4 with an arrow and a reference sign "L". The through hole 260 is configured to allow the fluid to pass through the hole. For example the through hole 260 can be configured to allow a tube, e.g. a sparge tube 220, containing the fluid to pass through the hole 260. Alternatively, a rod or similar passes through the hole 260 to make the construction of the tubing

arrangement stable.

Fig. 5A shows a side view of the body member 210 illustrated in Fig. 4 including the through hole 260 and a barb fitting 250. Fig. 5B shows a top view of the body member 210 illustrated in Fig. 4, wherein the through hole is illustrated to extend through the hole body member in a direction perpendicular to the length direction L of the body member.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims, the invention may be practiced otherwise than as specifically described. In addition, any combination of two or more of the features presented above is included within the scope of the present invention.

Claims

1 . A container comprising a tubing arrangement (200), the container (1 18) capable of holding a fluid and comprising a container wall (218) having an outer wall surface and an inner wall surface, the inner wall surface defining an interior chamber for holding the fluid, and the container (1 18) comprising at least two ports (230) configured to allow a fluid to flow into the interior chamber, wherein the at least two ports (230) are provided in the container wall (218), wherein the arrangement (200) comprises at least two sparge tubes (220) to provide the fluid in the interior chamber, and wherein each of the sparge tubes (220),

respectively, is connected to a port (230) in the interior chamber, and wherein the arrangement (200) comprises at least one body member (210) which couples the at least two sparge tubes (220) to keep the sparge tubes (220) in a fixed location.

2. The container according to claim 1 , wherein the tubing arrangement (200) further comprises at least one fastening element (232) fixed to the container wall (218) and arranged to be attached to the body member (210).

3. The container according to claim 1 or 2, wherein the tubing arrangement (200) comprises at least two body members (210), wherein each of the body members is configured to keep at least two sparge tubes (220) in a fixed location.

4. The container according to claim 3, wherein the at least two body members

(210) are connected to each other, whereby the sparge tubes (220) in the tubing arrangement (200) are kept in a fixed location.

5. The container according to any one of claims 1 to 4, wherein container

comprises a mixer (151 ) comprising an axially extending mixer shaft (157) and radially extending impeller blades (155) and the tubing arrangement (200) comprises at least four ports (230) provided in the container wall (218), wherein the ports (230) are connected to a respective sparge tube (220), and wherein the ports (230) radially surround the mixer (151 ).

6. The container according to any one of the preceding claims, wherein the body member (210) comprises a through hole (260) that extends perpendicular to the lengthwise direction (L) of the body member (210).

7. The container according to claim 6, wherein the through hole (260) is configured to allow the fluid to pass through the hole.

8. The container according to any one of the preceding claims, wherein the sparge tube (220) comprises a tee fitting (240) to direct the fluid to a desired direction.

9. The container according to any one of the preceding claims 1 -7, wherein the sparge tube (220) comprises a series of orifices to direct the fluid to a desired direction

10. The container according to any one of the preceding claims, wherein the sparge tube (220) comprises an end cap (222).

1 1 . The container according to any one of the preceding claims, wherein the body member (210) comprises at least one hose barb fitting (250) protruding outwards from the body member (210) for attaching the body member (210) to a further tube.

12. The container according to any one of the preceding claims, wherein the port (230) comprises a hose barb fitting for attaching the sparge tube (220) to the port (230).

13. The container according to any one of the preceding claims, wherein the at least one body member (210) is a rigid body member.

14. The container according to any one of the preceding claims, wherein the

container (1 18) is flexible.

15. A mixing vessel comprising the container according to any one of the preceding claims 1 -14.

16. A bioreactor comprising the container according to any one of the preceding claims 1 -14.