Large-scale bioreactor systems and methods

Abstract

The large-scale bioreactor system includes a stainless steel large-scale bioreactor having at least one valve assembly and a sterile connector assembly coupled to the at least one valve assembly of the bioreactor. The perfusion device includes an alternating tangential flow filtration assembly having an autoclaved valve assembly coupled to the sterile connector assembly, and the sterile connector assembly includes one of a tri-clamp sterile connector or a hose assembly. The single-use supply container includes the sterile connector assembly.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of the filing date of U.S. Provisional Patent Application No. 63 / 404,033, filed September 6, 2022, the entire contents of which are expressly incorporated herein by reference.

[0002] The present disclosure relates generally to large-scale bioreactor systems, and more particularly to the integration of perfusion devices and / or single-use supply vessels with large-scale bioreactors and associated pressure management. [Background technology]

[0003] The use of perfusion in cell culture bioreactors allows for improved performance compared to traditional fed-batch cell culture processes. Perfusion allows for the continuous addition of nutrients to the cell culture and the continuous removal of metabolic by-products. In fed-batch mode, the tank volume limits the cell culture process, and all by-products are contained inside the bioreactor until harvest.

[0004] Traditionally, perfusion has been applied to small-scale bioreactors, e.g., with a working volume of up to 2,000 L. The pressures realized by perfusion devices are greater on large-scale stainless steel bioreactors due to the increased height of liquid above the perfusion device (static head) when placed in its conventional location near the bottom of the side of a large-scale bioreactor. Additionally, perfusion devices containing single-use components are particularly sensitive to pressure due to the lower pressure ratings of the single-use components. Summary of the Invention [Means for solving the problem]

[0005] According to a first aspect, a large-scale bioreactor system includes a stainless steel large-scale bioreactor having at least one valve assembly and a sterile connector assembly coupled to the at least one valve assembly of the bioreactor. A perfusion device including an alternating tangential flow filtration (ATF) assembly having an autoclaved valve assembly is coupled to the sterile connector assembly, and the sterile connector assembly includes one of a tri-clamp sterile connector or a hose assembly.

[0006] According to a second aspect, a large-scale bioreactor system includes a stainless steel large-scale bioreactor having a side and an autoclaved valve assembly coupled to the side of the bioreactor, at least one sterile connector is coupled to the autoclaved valve assembly, and an irradiated single-use perfusion device is coupled to the at least one sterile connector.

[0007] According to yet another aspect, a large-scale bioreactor system includes a stainless steel large-scale bioreactor having at least one valve assembly and a single-use adapter assembly including a wye connector assembly coupled to the at least one valve assembly. Multiple single-use perfusion devices are connected to the single-use adapter assembly, allowing multiple perfusion units to be coupled to the bioreactor without the need for stationary steaming of the bioreactor when coupling multiple single-use perfusion units.

[0008] According to yet another aspect, a large-scale bioreactor system includes a stainless steel large-scale bioreactor and at least one stainless steel transfer panel having multiple inputs coupled to the bioreactor. Multiple single-use supply vessels are coupled to the at least one stainless steel transfer panel at the operating level of the bioreactor. Another aspect is that the stainless steel large-scale bioreactor can be installed in a pit recess in the floor to facilitate easy access to the probe belt for operation. Another aspect is that a completely enclosed stainless steel large-scale bioreactor can be installed in an uncontrolled space and operated with only a probe belt and localized additional panels. Such a configuration reduces the size of the controlled space, leading to lower operating costs for the manufacturing facility housing the bioreactor.

[0009] According to yet another aspect, a method of integrating at least one single-use perfusion device with a stainless steel large-scale bioreactor includes one of: (1) coupling a connector assembly to at least one valve assembly of the stainless steel large-scale bioreactor; or (2) coupling an autoclaved valve assembly to a side of the stainless steel large-scale bioreactor at a higher elevation to reduce pressure from a static head of liquid. The method further includes one of: (1) coupling an ATF assembly-autoclaved valve assembly of the single-use perfusion device to the connector assembly; or (2) coupling an irradiated single-use perfusion device to the autoclaved valve assembly. The method still further includes managing pressure in the single-use perfusion device via at least one pressure sensor in the single-use perfusion device, and automatically reducing, by a control system, one or more of the flow rate or pressure in the single-use perfusion device upon detecting a pressure above a safety limit pressure.

[0010] In some embodiments of any aspect, the stainless steel large scale bioreactor is configured to hold a volume of more than 2,000 L, such as a volume ranging from more than 2,000 L to 20,000 L, such as a volume ranging from 10,000 L to 20,000 L.

[0011] The present disclosure will be more fully understood from the following description taken in conjunction with the accompanying drawings. Some of the drawings may be simplified by the omission of selected elements for the purpose of more clearly showing other elements. The omission of such elements in some of the drawings does not necessarily indicate the presence or absence of the particular element in any of the illustrative embodiments, unless explicitly depicted in the corresponding written description. None of the drawings are necessarily drawn to scale. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 is a schematic diagram of a stationary steaming in place (SIP) bioreactor (BRX) with valve assemblies of a large-scale stainless steel bioreactor system according to one embodiment of the present disclosure, illustrating how the stationary steam bioreactor can be steamed in place without a perfusion device or a supply vessel connected to a valve. [Figure 2] FIG. 2 is a schematic diagram of the stationary steam bioreactor of FIG. 1 with a sterile connector assembly coupled to a valve assembly of the stationary steam bioreactor. [Figure 3] FIG. 3 is a schematic diagram of the stationary steam bioreactor of FIG. 2 with a connector assembly coupled to the valve assembly so that steaming can be performed in place, independent of the operation of the stationary steam bioreactor. [Figure 4] FIG. 4 is a schematic diagram of the stationary steam bioreactor of FIG. 2 or 3 with a single-use (SU) or reusable perfusion device including an alternating tangential flow filtration (ATF) assembly coupled to a connector assembly using a sterile connector. [Figure 5] FIG. 5 is an enlarged view of the perfusion device and associated valve assembly of FIG. 4 after it has been pre-autoclaved (COP) and assembled in preparation for connection to a stationary steam bioreactor. [Figure 6]FIG. 10 is another large-scale bioreactor system according to another embodiment of the present disclosure, having one of a plurality of autoclaved valve assemblies coupled to a large-scale stainless steel bioreactor and an irradiated single-use perfusion device aseptically coupled to the autoclaved valve assembly using a sterile connector. [Figure 7] FIG. 10 is another large-scale bioreactor system according to another embodiment of the present disclosure, including a hose assembly connecting at least one valve assembly of the stainless steel large-scale bioreactor to an autoclave perfusion device, such as a single-use perfusion device or a reusable autoclave perfusion device, including an ATF assembly. [Figure 8] FIG. 1 is a schematic diagram of a clean-in-place bioreactor having at least one valve assembly and the ability to clean the at least one valve assembly in-place using the clean-in-place bioreactor. [Figure 9] FIG. 10 is another large-scale bioreactor system according to another embodiment of the present disclosure, including a large-scale stainless steel bioreactor having at least one valve assembly and a wye assembly coupled to the at least one valve assembly to allow for the use of multiple perfusion devices on a single port. [Figure 10] FIG. 12 is a schematic diagram of multiple wye assemblies coupled to multiple valve assemblies, allowing multiple single-use supply vessels to be coupled to the wye assemblies and ultimately to the large-scale stainless steel bioreactor without the need to steam-in-place the valve assemblies multiple times. [Figure 11] FIG. 1 is a schematic diagram of a large-scale stainless steel bioreactor system having one of a plurality of autoclaved valve assemblies coupled to the top of the large-scale stainless steel bioreactor and an irradiated perfusion device aseptically coupled to at least one autoclaved valve assembly. [Figure 12]FIG. 1 is a schematic diagram of any one of the large-scale stainless steel bioreactors of the large-scale bioreactor system of the present disclosure, positioned at least partially within a pit to allow efficient operation of the probe belt and transfer panel. [Figure 13] FIG. 1 is a schematic diagram of a large-scale stainless steel bioreactor with a transfer panel coupled to the bioreactor that allows multiple feed vessels to be coupled to the stainless steel bioreactor at the operational level. DETAILED DESCRIPTION OF THE INVENTION

[0013] Generally, a large-scale bioreactor system is disclosed. The large-scale bioreactor system includes a stainless steel large-scale bioreactor having at least one valve assembly, a sterile connector assembly coupled to the at least one valve assembly of the bioreactor, and a single-use or reusable perfusion device coupled to the sterile connector assembly. The single-use perfusion device includes a perfusion filter assembly having an autoclaved valve assembly coupled to the sterile connector assembly. So configured, a new perfusion device can be installed while the stainless steel large-scale bioreactor is running cell culture by repeating stationary steaming of the sterile connector assembly or through use of the sterile connector valve assembly.

[0014] Referring now to FIGS. 1-5, a schematic diagram of a large-scale bioreactor system 10 (FIG. 4) according to some embodiments of the present disclosure is shown. As shown in FIG. 1, the large-scale bioreactor system 10 includes a stainless steel large-scale bioreactor 12 having at least one valve assembly 14. In these examples, the stainless steel large-scale bioreactor 12 is a stationary steam SIP bioreactor BRX having a large-scale volume. For example, the large-scale bioreactor 12 is configured to hold a volume of fluid up to 10,000 L, or in one example, more. The at least one valve assembly 14 of the stationary steam large-scale bioreactor 12 allows the stationary steam stainless steel large-scale bioreactor 12 to be fully sterilized before perfusion begins. Specifically, the at least one valve assembly 14 includes a first valve 16, a second valve 18, a third valve 20, and a fourth valve 21 disposed downstream from each of the first and second valves 16 and 18. Steam, such as clean steam CS, flows upward into third valve 20 and through first and second valves 16, 18 to port 24. After flowing into third valve 20, the steam also flows downward into fourth valve 21 through steam trap 22, where clean steam condensate CSC is released, for example, to sterilize bioreactor 12 and create a steam-sterilized, sterile environment. In one example, upon completion of a SIP cycle, condensate, such as clean steam condensate CSC, is discharged from steam trap 22. Additionally, as will be understood by those skilled in the art, process waste PW flows out of third and fourth valves 20, 21 and into a drain during cleaning. After use, stationary steam bioreactor 12 is cleaned in place and the stationary steam treatment process occurs again before any further use. At least one valve assembly 14 also includes a port 24 for coupling to a sterile connector valve assembly 26, such as an autoclaved sterile connector valve assembly 26 as shown in FIG. 2, or another device as further described below.

[0015] 2, a sterile connector valve assembly 26 is coupled to at least one valve assembly 14. Specifically, in this example, the sterile connector valve assembly 26 is coupled to the port 24 of the at least one valve assembly 14 and includes a first valve 28 and a second valve 30 disposed downstream from the first valve 28. A steam trap 32 is disposed further downstream from the second valve 30. In this example, the autoclaved connector valve assembly 26 also includes a tri-clamp sterile connector 34 coupled to the first valve 28. The sterile connector valve assembly 26 enables sterile connectivity of single-use or reusable perfusion devices to the large-scale bioreactor 12. The tri-clamp sterile connector 34 is configured to couple to a perfusion device, as described further below. As will be appreciated, in one example, the tri-clamp sterile connector 34 and the first and second valves 28, 30 may be pre-assembled and autoclaved prior to connection to the at least one valve assembly 14, e.g., for steam sterilization.

[0016] 3, the large-scale bioreactor 12 is again shown having at least one valve assembly 14 and a sterile connector valve assembly 26 coupled to the at least one valve assembly 14. However, in this example, the sterile connector valve assembly 26 is sterilized independently of the stationary steam large-scale bioreactor 12. Specifically, if, during a cell culture run in the large-scale bioreactor 12, it is determined that a perfusion device needs to be maintained and / or replaced, for example, the run of the large-scale bioreactor 12 can continue because the sterile connector valve assembly 26 can steam, for example, the first valve 28 and the second valve 30 and all other components of the sterile connector valve assembly 26 to provide a sterilized connector assembly for coupling to a repaired and / or new perfusion device.

[0017] Referring now to FIG. 4, a schematic diagram of a large-scale bioreactor system 10 according to one embodiment of the present disclosure is shown. More specifically, a stainless steel large-scale bioreactor 12 is shown having at least one valve assembly 14 and a sterile connector valve assembly 26 coupled to the at least one valve assembly 14. Additionally, a perfusion device 36, which may be a reusable or single-use perfusion device, is coupled to a tri-clamp sterile connector 34 of the sterile connector valve assembly 26. As shown in both FIG. 4 and the enlarged view of the perfusion device 36 in FIG. 5, the perfusion device 36 includes an alternating tangential flow filtration (ATF) assembly 38 having an autoclaved valve assembly 40 operably coupled to the sterile connector 34 of the sterile connector valve assembly 26. So configured, the large-scale bioreactor system 10 enables connectivity of a perfusion device 36, such as a reusable or single-use perfusion device, to the stainless steel large-scale bioreactor 12 without additional steam sterilization.

[0018] As also shown in FIG. 5 , the autoclaved valve assembly 40 of the perfusion device 36 includes a first valve 42 and a second valve 44 disposed downstream from the first valve 42. In this example, the autoclaved valve assembly 40 is cleaned out of situ (COP) and assembled prior to autoclaving in preparation for connection to the stationary steam bioreactor 12. COP generally refers to systems and equipment, such as the autoclaved valve assembly 40 in this example, that have undergone one or more of disassembly, rearrangement, or special processing to clean and sterilize. In other words, COP is defined as a method of cleaning equipment items by removing them from their operating area and transporting them to a designated cleaning station for cleaning. Additionally, as shown in FIG. 4 , a sterile connector 46 is operably coupled to the first valve 42 and directly coupled to the tri-clamp sterile connector 34 of the sterile connector assembly 26. The sterile connector assembly 26 allows connectivity between the single-use perfusion device 36 and the stainless steel large-scale bioreactor 12 and provides the ability to re-steam on subsequent perfusion devices during a single run of the bioreactor 12. In such an alternative, with this configuration, the perfusion device 36 and / or new perfusion devices can be installed by repeating the in-place steaming or using the sterile connector assembly 26, for example, while the stainless steel large-scale bioreactor 12 is running a cell culture.

[0019] Referring now to Figure 6, another large-scale bioreactor system 100 is shown. Similar to the large-scale bioreactor system 10 of Figures 1-5, the large-scale bioreactor system also includes a stainless steel large-scale bioreactor 112 having a side 113 and a first valve assembly 114, such as an autoclaved valve assembly 114, coupled to the side 113 of the stainless steel large-scale bioreactor 112. The autoclaved valve assembly 114 includes a first valve 116, a second valve 118 downstream from the first valve 116, a third valve 120 downstream from both the first valve 116 and the second valve 118, and a fourth valve 121 downstream from the third valve 120. Again, similar to the valve assembly 14 of the system 10 of Figures 1-5, steam, such as clean steam CS, flows into the third valve 120, upward through the first and second valves 116, 118, and into port 134. Furthermore, after flowing into the third valve 120, the steam flows downward through a fourth valve 121 and through a steam trap 122 to sterilize the bioreactor 112, e.g., to allow for an in-place steaming process to occur prior to further use. However, in this large-scale bioreactor system 100, similar to the sterile connector assembly 26 of the system 10 of FIGS. 1-5, there is only at least one sterile connector 134 coupled to the autoclaved valve assembly 114, such as the first valve 116 of the autoclaved valve assembly 114, and not the second valve assembly. This same autoclaved valve assembly 114 allows for connection via the at least one sterile connector 134 of the autoclaved valve assembly 114 to a factory-assembled and irradiated perfusion device 136, which may be, for example, a single-use or reusable perfusion device. More specifically, as shown in FIG. 6, the irradiated perfusion device 136 includes a first sterile connector 138 coupled to the at least one sterile connector 134 .Irradiated perfusion device 136 also includes a second sterile connector 140 configured to be coupled to a second autoclaved valve assembly (not shown), such as a second valve assembly, which is also configured to be directly coupled to side 113 of large scale stainless steel bioreactor 112 on a different valve assembly similar to valve assembly 114. As a result, multiple perfusion devices can be aseptically coupled to large scale stainless steel bioreactor 112 without additional steam sterilization.

[0020] Referring now to Figure 7, a schematic diagram of another large-scale bioreactor system 200 of the present disclosure is shown. Similar to the large-scale bioreactor system 10 of Figure 4, for example, the large-scale bioreactor system 200 includes a stainless steel large-scale bioreactor 212 having a sidewall 213 and at least one valve assembly 214 disposed on the sidewall 213. Again similar to the large-scale bioreactor system 10 of Figure 4, the same valve assembly 214 in the large-scale bioreactor 200 of Figure 7 allows for the connection of an autoclaved device and cleaning-in-place of the valve assembly 214, but without the use of a sterile connector such as the sterile connector 34 of Figure 4. Instead, a hose assembly is used as an alternative connector for coupling the stainless steel large-scale bioreactor 212 to an autoclaved perfusion device, as described further below.

[0021] More specifically, as shown in FIG. 7 , the stainless steel large-scale bioreactor 212 of the large-scale bioreactor system 200 is also a stationary steam bioreactor having a large-scale volume. For example, the large-scale bioreactor 212 is configured to hold a volume of fluid of up to 10,000 L, or in one example, greater than 10,000 L. The at least one valve assembly 214 of the stationary steam large-scale bioreactor 212 allows the stationary steam stainless steel large-scale bioreactor 212 to be fully sterilized once perfusion begins. Specifically, similar to the at least one valve assembly 14 of the bioreactor system 10 of FIG. 4 , the at least one valve assembly 214 includes a first valve 216, a second valve 218, and a third valve 220 located downstream from each of the first and second valves 216, 218. As shown, a fourth valve 221 may also be located downstream from the third valve 220. Steam, such as clean steam CS, again flows upward into third valve 220 and through first and second valves 116, 118 to port 224. Furthermore, after flowing into third valve 220, the steam also flows downward into fourth valve 221 and through steam trap 222 to sterilize bioreactor 212 and create a steam-sterilized, sterile environment. Furthermore, as will be appreciated by those skilled in the art, process waste PW may again flow out of third and fourth valves 220, 221 and be discharged into a drain during cleaning. After use, stationary steam bioreactor 212 is cleaned in place and again undergoes a stationary steaming process before any further use. At least one valve assembly 214 also includes a port 224 for connection to hose assembly 235, which in turn connects to a perfusion device or another device as further described below.

[0022] The hose assembly 235 includes a hose body 237 having a first end 237A and a second end 237B. The first end 237A is removably coupled to the port 224 of the at least one valve assembly 214, and the second end 237B is coupled to an autoclaved perfusion device 236, which is functionally equivalent to the perfusion device 36 of the system 10 of FIG. 4 but does not have a single-use connector, such as the single-use connector 46 of FIG. 5. The autoclaved perfusion device 236 may include a single-use or reusable perfusion device and still be within the scope of the present disclosure. The autoclaved perfusion device 236 includes an ATF assembly 238 having an autoclaved valve assembly 240 coupled to the second end 237b of the hose body 237. The autoclaved valve assembly 240 of the autoclaved perfusion device 236 includes a first valve 242 and a second valve 244 disposed downstream from the first valve 242. The second end 237B of the hose body 237 is operably coupled to the connector of the first valve 242. The hose assembly 235 enables connectivity between the autoclaved device 236 and the stainless steel large-scale bioreactor 212 and provides the ability to resteam on a subsequent perfusion device during a single run of the stainless steel large-scale bioreactor 212.

[0023] 8 , hose assembly 235 can be disconnected from its connection to perfusion device 236 and coupled to another one of multiple ports 224 on the same large-scale bioreactor 212. Stationary purification valve assembly 250 allows at least one valve assembly 214 to be cleaned, thereby allowing another perfusion device, such as a single-use perfusion device, to be aseptically coupled to the stainless steel large-scale bioreactor 212 via hose assembly 235. In this example, the stationary purification steam supply of stationary purification valve assembly 250 flows upward through hose assembly 235, through first valve 216 into third valve 220 and second valve 218 of at least one valve assembly 214, to port 224. In addition, the stationary purification steam supply also flows downward from third valve 220 through downstream fourth valve 221. In this manner, each of the first, second, third, and fourth valves 216, 218, 220, and 221 of the at least one valve assembly 214 and port 224 is cleaned. While this description refers to at least one valve assembly 214, it will be understood that the at least one valve assembly 214 may include multiple valve assemblies (not shown), and the stationary clean steam supply of the stationary clean valve assembly 250 may also flow through each of the valves and ports of another one of the multiple valve assemblies, allowing multiple valve assemblies to be cleaned.

[0024] 9, another schematic diagram of another large-scale bioreactor system 300 of the present disclosure is shown. The large-scale bioreactor system 300 includes a stainless steel large-scale bioreactor 312 having a side 313 and at least one valve assembly 314 coupled to the side 313, and an autoclaved valve assembly 326 coupled to the at least one valve assembly 314. In one example, the stainless steel large-scale bioreactor 312 is again a stationary steam bioreactor having a large volume. For example, the large-scale bioreactor 312 is configured to hold a volume of fluid up to 10,000 L, or in one example, greater than 10,000 L. The at least one valve assembly 314 of the stationary steam large-scale bioreactor 312 allows the stationary steam stainless steel large-scale bioreactor 312 to be fully sterilized once perfusion begins. 4, for example, the at least one valve assembly 314 includes a first valve 316, a second valve 318, a third valve 320 disposed downstream from each of the first and second valves 316, 318, and a fourth valve 321 disposed downstream from the third valve 320. Again, steam, such as clean steam CS, flows upward into the third valve 320 and into the second and first valves 318, 316 to port 324 to sterilize the at least one valve assembly 314 and the bioreactor 312. Additionally, after flowing through the third valve 320, the steam also flows downward through a fourth valve 321 and a steam trap 322 to sterilize the bioreactor 312 and create a steam-sterilized, sterile environment. Additionally, as will be understood by those skilled in the art, process waste PW flows out of the fourth valve 321 and into a drain during cleaning. After use, the steam-in-place bioreactor 312 is cleaned-in-place and the steaming process occurs again before any further use. The at least one valve assembly 314 again includes a port 324 for coupling to an autoclaved valve assembly 326 and a sterile connector 334.

[0025] Unlike the other bioreactor systems 10, 100, and 200, bioreactor system 300 includes an adapter assembly 360, such as a single-use adapter assembly and / or a wye assembly, that is configured to be and / or be coupled to a sterile connector 334, as also shown in Figure 9. So configured, wye assembly 360 is coupled to at least one valve assembly 314, in this example, via sterile connector 334.

[0026] An adapter assembly 360, such as wye assembly 360, includes a connector 361 that couples directly to sterile connector 334 (which is ultimately coupled to port 324), a pair of tubes 362 extending outward from connector 361, a first sterile connector 364 coupled to one of the tubes 362, and a second sterile connector 366 coupled to the other tube 362. Each of the first sterile connector 364 and second sterile connector 366 of wye assembly 360 is configured to couple to a reusable or single-use perfusion device, such as any one of the previously described perfusion devices 36, 136, and 236. This allows, for example, multiple perfusion devices to be operably coupled to stainless steel large-scale bioreactor 312 via port 324 of valve assembly 314 without the need for stationary steam treatment of stainless steel large-scale bioreactor 312 when coupling multiple reusable or single-use perfusion devices. In another example, an adapter assembly 360, such as wye assembly 360, can be a first wye assembly 360, and a second wye assembly (not shown) can be coupled to the first wye assembly. So configured, coupling one or more additional wye assemblies to the first wye assembly 360 allows two or more reusable or single-use perfusion devices and multiple reusable or single-use perfusion devices to be operably coupled to the stainless steel large-scale bioreactor 312, again without the need for stationary steaming of the stainless steel large-scale bioreactor 312 when coupling multiple single-use perfusion devices. Similarly, a greater number of perfusion devices can be connected to the large-scale bioreactor 312, each without the need for a separate reactor port.

[0027] 10, in another example, a large-scale bioreactor system 300 is shown that includes a large-scale stainless steel bioreactor 312 configured to hold a volume of fluid up to, or in one example, greater than, 10,000 L. In this example, the at least one valve assembly of the large-scale bioreactor system 300 includes at least one valve assembly 380 configured to be coupled to a transfer panel (not shown in FIG. 10) mounted at a local or remote distance from the large-scale stainless steel bioreactor 312 for the purpose of connecting and transferring a feed liquid held in a single-use container to the large-scale stainless steel bioreactor 312. The valve assembly 380 is configured to allow steam sterilization of a transfer line 385 and connection to a single-use supply container. Steam from the large-scale bioreactor 300, in one example, exits through at least one transfer line, such as transfer line 385, passes downward to a tri-clamp port 324 and a sterile connector assembly 334 through a first valve 381, a second valve 382, ​​a third valve 383, and a fourth valve 384 to the steam trap 322. Thus, the entire line is made sterile up to the sterile connector 334. Similarly, cleaning of the line is accomplished by supplying a clean-in-place (CIP) solution through the first and second valves 381, 382, ​​through the transfer line 385 to the large-scale bioreactor 300. As shown in FIG. 10 , ports 324 of the first and second valves 381, 382 are coupled to a single-use adapter, such as the sterile connector 334, with the port 324 designed to allow direct exposure to clean steam at sterilization temperatures and pressures. As further shown, single-use adapters, such as sterile connector assemblies 334, are then coupled to single-use manifolds 370. In this example, each single-use manifold 370 includes eight or more arms 371 having inlets 372, each configured to couple to a supply container 373, such as a single-use supply container. In another example, single-use manifold 370 can include more or fewer than eight arms 371, such as four or six, and still be within the scope of the present disclosure.So configured, multiple supply vessels can be operably coupled to the stainless steel large-scale bioreactor 312 without additional steaming and cleaning. One or more additional supply vessels 373 can be connected using the sterile connectors 372 during operation of the large-scale stainless steel bioreactor 312. Additionally, one or more additional single-use manifolds 370 can be connected to the inlet 372 via sterile connectors on the first single-use manifold assembly 370 to further increase the number of supply vessels 373 that can be connected.

[0028] Referring again to FIG. 10 , if a sterile connector is used that cannot tolerate direct exposure to clean steam at sterilization temperatures and pressures, another configuration allows for isolating the sterile connector from the steam. For example, one or more valve assemblies 390A and 390B can be prepared by combining a valve or two valves, such as valve 391A of valve assembly 390A and first and second valves 391B and 393B of valve assembly 390B, which can be any valve with a low holding volume on its side so that it can be steamed and autoclaved relative to a sterile connector, such as sterile connector 392A or sterile connector 392B. Each of valve assemblies 390A and 390B can be inserted, for example, by combining with a tri-clamp connector, into a valve assembly that can be installed in a local or remote transfer panel, such as at least one valve assembly 380A shown in FIG. 10 . From that point on, cleaning, steaming, and operation are the same as those described above with respect to FIG. 9 . The use of a single-use manifold assembly with sterile connectors 392 provides a larger inner diameter, lower shear, and higher flow rates than can be achieved using sterile connectors that can be directly exposed to clean steam at sterilization temperatures and pressures. However, because the main transfer line 385 and at least one valve assembly 380, 380a are the same, either type of sterile connector can be used and multiple similar supply vessels 373 can be connected. Alternatively, if higher flow rates or lower shear are required, a single connection to the supply vessel 373 can be used.

[0029] 11, the bioreactor system 100 of FIG. 6 is again shown, including all of the same features of the bioreactor system 100, except for the location of the valve assembly 114 relative to the stainless steel large-scale bioreactor 112. Specifically, the valve assembly 114 is mounted at a higher position on the side 113 of the stainless steel large-scale bioreactor 112, which in turn causes the perfusion device 136 to be mounted at a higher position on the side 113 of the stainless steel large-scale bioreactor 112. Notably, in this example, the side 113 of the bioreactor 112 includes a bottom end 113A and a top end 113B, and the valve assembly 114 is coupled to the side 113 of the bioreactor at a position closer to the top end 113B of the side 113 than to the bottom end 113A, as shown in FIG. 11. In this configuration, the height, and therefore the pressure, of the liquid L disposed above the perfusion device 136 is reduced.

[0030] Generally, when installed in a typical location near the bottom, such as the bottom end 113A, of a stainless steel large-scale bioreactor 12, 112, 136, 236, the pressure achieved by a single-use perfusion device, such as the perfusion device 36, 136, 236, will be greater on the stainless steel large-scale bioreactor 12, 112, 212, 312 due to the increased height of liquid (static head) above the perfusion device 36, 212, 312. Perfusion devices that include single-use components are particularly sensitive to greater pressures due, for example, to the lower pressure ratings of the single-use components. Therefore, installing the valve assembly 114, and thus the perfusion device 136 of FIG. 11, reduces pressure near the top end 113B of the bioreactor 112.

[0031] 11 , a perfusion device 136, such as a single-use perfusion device, includes at least one pressure sensor 180 and may include additional pressure sensors. In this example, the perfusion device 136, such as an RFT single-use perfusion device, includes a first pressure sensor 180, a second pressure sensor 182, and a third pressure sensor 184. In addition, the perfusion device 136 also includes a circulation pump 186 disposed upstream from the first pressure sensor 180 and a permeate pump 188 disposed downstream from the third sensor 184.

[0032] 11 , a control system 190 for monitoring the pressure of at least one perfusion device 136 is communicatively coupled to the single-use perfusion device 136 via a network, such as a wireless network 192 or a wired connection 193. While the control system 190 is operatively coupled to the perfusion device 136, the control system 190 may optionally be coupled to any one of the other aforementioned perfusion devices 36, 236 and still fall within the scope of the present disclosure. In one example, the control system 190 includes a computing device 194 having a memory 195, a processor 196, and a network interface 197. It will be understood that the computing device 194 may include any known inputs, receivers, and transmitters. Additionally, the control system 190 further includes an alarm 198. So configured, the control system 190 monitors the pressure at one or more of the first, second, and third pressure sensors 180, 182, and 184. If the pressure approaches a predefined safety limit, an alarm 198 will be activated to notify the user so that the bioreactor system 100 can be investigated and evaluated. If the pressure monitored by the control system 190 approaches a predefined safety limit any further, the control system 190 will automatically reduce the fluid flow rate, and therefore the pressure, to maintain a safe operating pressure of the bioreactor system 100. If the pressure monitored by the control system 190 reaches a safe pressure limit, the control system 190 will stop the circulation pump 186 to prevent damage to the perfusion device 136.

[0033] 12, any one of the large scale stainless steel bioreactors 12, 112, 212, 312 described above can be placed in a pit 402 as shown in FIG. 12. Additionally, a valve assembly 14, 114, 214, 314 is then attached near the bottom end 13A, 113A, 213A, 313A of the side 13, 113, 213, 313 of the large scale stainless steel bioreactor 12, 112, 212, 312 and to a bottom weld seam 404 on the body of the bioreactor 12, 112, 212, 312. This allows the single-use perfusion device 36, 136, 236 to be placed at a higher position and minimizes the hold-up volume between the perfusion device 36, 136, 236 and the large scale stainless steel bioreactor 12, 112, 212, 312. Alternatively, by adjusting the depth of the pit, the pit 402 may be used to position any one of the perfusion devices 36, 136, 236 higher on the side 13, 113, 213 of the bioreactor 12, 112, 212, thereby also reducing pressure.

[0034] Referring now to FIG. 13 , another schematic diagram of another large-scale bioreactor system 500 of the present disclosure is shown. The large-scale bioreactor system 500 includes a stainless steel large-scale bioreactor 512 having a side 513 with a bottom end 513A and a top end 513B. Additionally, at least one stainless steel transfer panel 520 having multiple inputs 522 is coupled to the side 513 of the stainless steel large-scale bioreactor 512 at the top end 513B of the side 513. Furthermore, at an operating level 526 of the stainless steel large-scale bioreactor 512, multiple single-use supply containers 524 are coupled to one or more of the multiple inputs 522 of the transfer panel 520. While one stainless steel transfer panel 520 is shown in FIG. 13 , it will be understood that multiple stainless steel transfer panels could alternatively and / or additionally be coupled to the bioreactor 512 and still fall within the scope of the present disclosure. So configured, this avoids the user having to climb stairs every time they want to connect something to the bioreactor 512. Additionally, this allows for connections to be made to single-use supply containers inside a smaller clean space, for example, with some portions of the bioreactor located in a controlled, non-classified (CNC) space, without placing the entire bioreactor 512 inside the clean space. This also allows a user to work from the clean, controlled space and operate the bioreactor 512 from the clean, controlled space without having to access the top end of the bioreactor 512.

[0035] Additionally, it will be appreciated that any components of any one or more of the aforementioned bioreactor systems 10, 100, 200, 300, and 500 requiring steaming may be steamed while the associated bioreactor 12, 112, 212, 312, 512 is being steamed. In this scenario, the steam source is from the bioreactor tank 12, 112, 312, 412, 512, and the steam flows through the valves to the steam traps shown. For example, in the bioreactor system 10 of FIG. 2, steam may flow from the bioreactor 12 through the first and second valves 16, 18 to the third and fourth valves 20, 21 and to the steam trap 22. Similarly, in the bioreactor system 100 of FIG. 6, steam may also flow from the bioreactor 112 through the first, second, third, and fourth valves 116, 118, 120, 121 to the steam trap 122. 9, steam can again flow from bioreactor 212 into first, second, third, and fourth valves 216, 218, 220, 221 to steam trap 222. Similarly, in bioreactor system 300, steam can again flow from bioreactor 312 into first and second valves 316, 318 through third and fourth valves 320, 321 to steam trap 322.

[0036] In view of the foregoing, it will be appreciated that any one or more of the aforementioned bioreactors 12, 112, 212, 312 may be integrated with any one or more of the aforementioned perfusion devices, such as reusable or single-use perfusion devices 36, 136, 236, according to one or more of the following methods. Specifically, according to one example, a method of integrating at least one single-use perfusion device 36, 136, 236 with a stainless steel large-scale bioreactor 12, 112, 212, 312 includes one of: (1) coupling a connector assembly 26, such as a sterile connector assembly 26, to at least one valve assembly 14, 214 of the stainless steel large-scale bioreactor 12, 112, 212, 312; or (2) coupling an autoclaved valve assembly 116 to the side 113 of the stainless steel large-scale bioreactor 112. Additionally, the method further includes one of (1) coupling an autoclaved valve assembly 40, 240 of the ATF assembly of the single-use perfusion device 36, 236 to the connector assembly 26, or (2) coupling the irradiated perfusion device 136 to the autoclaved valve assembly. Additionally, the method still further includes managing the pressure in the perfusion device 36, 136, 236 via at least one pressure sensor in the perfusion device 36, 136, 236, and automatically reducing one or more of the flow rate or pressure in the perfusion device 36, 136, 236 by the control system 190 upon detecting a pressure above a safety limit pressure.

[0037] In one example, the method includes coupling a sterile connector assembly to at least one valve assembly 14, 214 of the stainless steel large-scale bioreactor 12, 212 and coupling an autoclaved valve assembly 40 to a sterile connector assembly 26, where the sterile connector assembly 26 includes one of a tri-clamp connector assembly or a hose assembly. In another example, the method includes coupling an autoclaved valve assembly 114 to the side 113 of the stainless steel large-scale bioreactor 112 and coupling a perfusion device 136 to one of the plurality of autoclaved valve assemblies 114. While only one autoclaved valve assembly 114 is shown, it will be understood that more than one autoclaved valve assembly 114 can be coupled to the perfusion device 136 and still fall within the scope of the present disclosure.

[0038] In yet another example, the method includes coupling a connector assembly 360 to at least one valve assembly 314 of a stainless steel large-scale bioreactor 312, where the connector assembly 360 includes a wye connector assembly. Additionally, the method further includes coupling an autoclaved valve assembly 40 of an ATF assembly of at least one perfusion device 36 to the connector assembly 360, where the at least one perfusion device 36 or supply vessel 373, 524 includes multiple perfusion devices or supply vessels connected to the wye connector assembly. This allows multiple perfusion devices or supply vessels to be coupled to the bioreactor 12, 112, 212, 312 without having to steam-in-place the bioreactor 12, 112, 212, 312 when coupling the multiple perfusion devices or supply vessels.

[0039] The above description describes various bioreactor systems and methods that integrate at least one single-use perfusion device and / or at least one feed vessel with a stainless steel large-scale bioreactor. It will be appreciated that the systems and methods of the present disclosure include several advantages. For example, the described systems and methods allow for connection of a perfusion device to a large-scale bioreactor (e.g., a bioreactor having a volume of over 2,000 L), exchange of a perfusion device or feed vessel during a cell culture run, and pressure management of the perfusion device while creating a steam sterilization aseptic environment. Additionally, the same valve assembly in the bioreactor allows for both connection of factory-assembled, irradiated single-use perfusion devices and autoclaved devices, as well as in-place cleaning of the bioreactor valve assembly, for example, without the use of sterile connectors.

[0040] While the foregoing methods and systems and elements thereof have been described in terms of exemplary embodiments, they are not limited to these exemplary embodiments. The detailed description is to be construed as exemplary only and does not describe every possible embodiment of the invention, as describing every possible embodiment of the invention would be impractical, if not impossible. Many alternative embodiments can be implemented, using either current technology or technology developed after the filing date of this patent, that would still fall within the scope of the claims that define the invention.

[0041] It is understood that the legal scope of the present invention is defined by the terms of the claims set forth at the end of this patent. The appended claims should be interpreted broadly to include other variations and embodiments thereof that may be made by those skilled in the art without departing from the scope of equivalents of the devices, systems, methods, and elements thereof.

Claims

1. A large-scale bioreactor system, A large stainless steel bioreactor having at least one valve assembly, A sterile connector assembly coupled to the at least one valve assembly of the bioreactor, A perfusion device or reusable alternating tangential flow filtration (ATF) assembly having an autoclaved valve assembly coupled to the sterile connector assembly, wherein the sterile connector assembly includes one of a Triclamp sterile connector or a hose assembly, and A large-scale bioreactor system including a bioreactor.

2. The bioreactor system according to claim 1, wherein the perfusion device is a single-use perfusion device, the single-use perfusion device further includes at least one pressure sensor, and the bioreactor system further includes a control system coupled to the single-use perfusion device for monitoring the pressure of the single-use perfusion device via the at least one pressure sensor.

3. The bioreactor system according to claim 1, wherein the perfusion device or reusable ATF assembly is a single-use perfusion device or reusable ATF assembly further comprising at least one pressure sensor, and the bioreactor system further comprising a control system coupled to the single-use perfusion device or the reusable ATF assembly for monitoring the pressure of the single-use perfusion device or the reusable ATF assembly via the at least one pressure sensor.

4. The bioreactor system according to claim 2 or 3, further comprising an alarm, the control system being configured to activate when the control system detects a pressure exceeding a safety pressure limit, and the control system being configured to automatically reduce one or more flow rates or pressures in the single-use perfusion device or the reusable ATF assembly in response to the detection of the pressure exceeding the safety pressure limit.

5. The bioreactor system according to any one of claims 1 to 3, wherein the bioreactor has a body with a side portion having an upper end, and at least one valve assembly of the bioreactor and one or more of the autoclaved valve assemblies are coupled to the side portion of the bioreactor near the upper portion of the bioreactor.

6. The bioreactor system according to any one of claims 1 to 3, wherein the bioreactor comprises a body and a bottom, the bioreactor is configured to be at least partially positioned within a pit, and at least one valve assembly of the stainless steel large bioreactor and one or more of the autoclaved valve assemblies are bonded to the bottom weld seam of the bottom of the bioreactor.

7. The bioreactor system according to any one of claims 1 to 3, wherein the sterile connector assembly is a sterile connector valve assembly having a Triclamp sterile connector, and a new perfusion device or ATF assembly is coupled to the stainless steel large bioreactor by repeatedly subjecting the sterile connector assembly to stationary steam treatment while the bioreactor is performing cell culture.

8. The bioreactor system according to any one of claims 1 to 3, wherein the sterile connector assembly is a hose assembly, the hose assembly connects the at least one valve assembly of the bioreactor to the perfusion device or ATF assembly, the hose assembly includes a hose body having a first end and a second end, the first end being operably coupled to the at least one valve assembly, and the second end being operably coupled to the perfusion device or ATF assembly.

9. A large-scale bioreactor system, A large-scale stainless steel bioreactor with sides, An autoclaved valve assembly coupled to the side of the bioreactor, At least one sterile connector coupled to the autoclaved valve assembly, A pre-irradiated, single-use perfusion device coupled to at least one sterile connector and A large-scale bioreactor system including a bioreactor.

10. The bioreactor system according to claim 9, wherein the autoclaved valve assembly is a first valve assembly, and the bioreactor system further comprises a second autoclaved valve assembly configured to be coupled to the side of the bioreactor adjacent to the first valve assembly, and at least one sterile connector configured to be coupled to the second autoclaved valve assembly.

11. The bioreactor system according to claim 9 or 10, wherein the irradiated single-use perfusion device further includes at least one pressure sensor, and the bioreactor system further includes a control system coupled to the irradiated single-use perfusion device for monitoring the pressure of the single-use perfusion device via the at least one pressure sensor.

12. The bioreactor system according to claim 11, further comprising an alarm, the alarm being configured to activate when the control system detects a pressure exceeding a safety pressure limit, and the control system being configured to automatically reduce one or more flow rates or pressures in the single-use perfusion device in response to the detection of the pressure exceeding the safety pressure limit.

13. The bioreactor system according to claim 9 or 10, wherein the bioreactor has a body with a side portion having an upper end, and the autoclaved valve assembly is coupled to the side portion of the bioreactor near the upper end of the bioreactor.

14. The bioreactor system according to claim 9 or 10, wherein the bioreactor has a body and a bottom end, the bioreactor is configured to be at least partially positioned in a pit, and the autoclaved valve assembly is bonded to the bottom weld seam of the bottom of the bioreactor.

15. A large-scale bioreactor system, A large stainless steel bioreactor having at least one valve assembly, A single-use adapter assembly including a wire connector assembly coupled to at least one valve assembly, A plurality of single-use perfusion devices connected to the single-use adapter assembly, wherein the plurality of perfusion units can be coupled to the bioreactor without requiring stationary steam treatment of the bioreactor when coupling the plurality of single-use perfusion units, and A large-scale bioreactor system including a bioreactor.

16. The bioreactor system according to claim 15, further comprising a single-use manifold coupled to one of the single-use adapter assemblies or one of the at least one valve assemblies, wherein the single-use manifold comprises any one of two, three, four, five, six, seven, or eight arms, each arm comprising an inlet for coupling to a single-use perfusion unit, enabling the plurality of single-use perfusion devices to be operably coupled to the stainless steel large bioreactor.

17. The bioreactor system according to claim 15, wherein the Wy connector assembly is a first Wy connector assembly, and a plurality of Wy connector assemblies are coupled to the first Wy connector assembly, enabling the plurality of single-use perfusion devices to be operably coupled to the stainless steel large bioreactor.

18. The bioreactor system according to any one of claims 15 to 17, wherein the single-use perfusion device further includes at least one pressure sensor, and the bioreactor system further includes a control system for monitoring the pressure of at least one of the plurality of single-use perfusion devices via the at least one pressure sensor.

19. The bioreactor system according to claim 18, further comprising an alarm, the control system being configured to activate when the control system detects a pressure exceeding a safety pressure limit, and the control system being configured to automatically reduce one or more flow rates or pressures in at least one of the plurality of single-use perfusion devices in response to the detection of the pressure exceeding the safety pressure limit.

20. The bioreactor system according to any one of claims 15 to 17, wherein the bioreactor has a body with a side portion having an upper end, and the at least one valve assembly of the bioreactor is coupled to the side portion of the bioreactor near the upper part of the bioreactor.

21. The bioreactor system according to any one of claims 15 to 17, wherein the bioreactor comprises a body and a bottom, the bioreactor is configured to be at least partially positioned within a pit, and the at least one valve assembly of the large stainless steel bioreactor is bonded to the bottom weld seam of the bottom of the bioreactor.

22. A large-scale bioreactor system, A large-scale stainless steel bioreactor, At least one stainless steel transfer panel having multiple inputs coupled to the bioreactor, At the operating level of the bioreactor, a plurality of supply containers coupled to the at least one stainless steel transfer panel and A large-scale bioreactor system including a bioreactor.

23. The bioreactor system according to claim 22, wherein the at least one stainless steel transfer panel is a first stainless steel transfer panel, and the bioreactor system further comprises one or more additional stainless steel transfer panels, each configured to be coupled to an additional plurality of supply containers.

24. The bioreactor system according to claim 22 or 23, wherein the plurality of supply containers further include at least one pressure sensor, and the bioreactor system further includes a control system for monitoring the pressure of at least one of the plurality of single-use supply containers via the at least one pressure sensor, wherein the at least one supply container is at least one single-use supply container.

25. The bioreactor system according to claim 24, further comprising an alarm, the control system being configured to activate when the control system detects a pressure exceeding a safety pressure limit, and the control system being configured to automatically reduce one or more flow rates or pressures in at least one of the plurality of single-use supply containers in response to the detection of the pressure exceeding the safety pressure limit.

26. The bioreactor system according to claim 22 or 23, wherein the bioreactor comprises a body having a side portion having an upper end, and at least one valve assembly coupled to the side portion of the bioreactor near the upper end of the bioreactor.

27. The bioreactor system according to claim 22 or 23, wherein the bioreactor comprises at least one valve assembly and a body with a bottom, the bioreactor is configured to be at least partially positioned in a pit, and the at least one valve assembly of the large stainless steel bioreactor is bonded to a bottom weld seam of the bottom of the bioreactor.

28. The large-scale stainless steel bioreactor has a volume of more than 2,000 L, as described in any one of claims 1 to 3.

29. The large-scale stainless steel bioreactor has a volume in the range of more than 2,000 L to 20,000 L, as described in any one of claims 1 to 3.

30. A method for integrating at least one perfusion device with a stainless steel large-scale bioreactor, (1) coupling the connector assembly to at least one valve assembly of the stainless steel large bioreactor, or (2) coupling the autoclaved valve assembly to the side of the stainless steel large bioreactor, (1) connecting the autoclaved valve assembly of the alternating tangential flow filtration (ATF) assembly of the perfusion device to the connector assembly, or (2) connecting the irradiated perfusion device to the autoclaved valve assembly, The pressure of the perfusion device is managed via at least one pressure sensor of the perfusion device, and the control system automatically reduces one or more of the flow rate or pressure in the perfusion device when it detects a pressure exceeding the safety limit pressure. A method that includes this.

31. The method according to claim 30, wherein one of (1) coupling a connector assembly to at least one valve assembly of a stainless steel large bioreactor, or (2) coupling an autoclaved valve assembly to the side of the stainless steel large bioreactor, is to couple a sterile connector valve assembly to the at least one valve assembly of the stainless steel large bioreactor.

32. The method according to claim 31, wherein one of (1) coupling an autoclaved valve assembly of an ATF assembly of a perfusion device to the connector assembly, or (2) coupling an irradiated perfusion device to the autoclaved assembly, comprises coupling the autoclaved valve assembly of the perfusion device to the connector assembly, the connector assembly comprising one of a triclamp connector assembly or a hose assembly.

33. The method according to claim 32, wherein one of (1) coupling a connector assembly to at least one valve assembly of a stainless steel large bioreactor, or (2) coupling an autoclaved valve assembly to the side of the stainless steel large bioreactor, is the autoclaved assembly to the side of the stainless steel large bioreactor.

34. The method according to claim 33, wherein one of (1) coupling an autoclaved valve assembly of a perfusion device ATF assembly to the connector assembly, or (2) coupling an irradiated perfusion device to the autoclaved valve assembly, comprises coupling the irradiated perfusion device to the autoclaved assembly.

35. The method according to claim 30, wherein one of (1) coupling a connector assembly to at least one valve assembly of a stainless steel large bioreactor, or (2) coupling an autoclaved valve assembly to the side of the stainless steel large bioreactor, comprises coupling a connector assembly to at least one valve assembly of the stainless steel large bioreactor, the connector assembly includes a Wy connector assembly.

36. The method according to claim 35, wherein one of (1) coupling an autoclaved valve assembly of an ATF assembly of at least one perfusion device to the connector assembly, or (2) coupling an irradiated perfusion device to at least one sterile connector comprises coupling the autoclaved valve assembly of an ATF assembly of at least one perfusion device to the connector assembly, wherein the at least one perfusion device comprises a plurality of perfusion devices connected to the connector assembly, and the plurality of perfusion devices can be coupled to the bioreactor without requiring the bioreactor to undergo stationary steam treatment when coupling the plurality of perfusion devices.

37. The method according to any one of claims 30 to 36, further comprising coupling at least one stainless steel transfer panel having multiple inputs to the stainless steel large bioreactor.

38. The method according to any one of claims 30 to 36, further comprising coupling the at least one valve assembly of the stainless steel large bioreactor and one or more of the autoclaved valve assemblies to the side of the stainless steel large bioreactor.

39. The stainless steel large-scale bioreactor has a volume of more than 2,000 L, according to any one of claims 30 to 36.

40. The method according to any one of claims 30 to 36, wherein the large-scale stainless steel bioreactor has a volume in the range of more than 2,000 L to 20,000 L.

Images