LARGE SCALE BIOREACTOR SYSTEM AND METHOD

A large scale bioreactor system includes a stainless steel large scale bioreactor having at least one valve assembly, and an aseptic connector assembly coupled to the at least one valve assembly of the bioreactor. A perfusion device includes an Alternating Tangential Filtration assembly with an autoclaved valve assembly coupled to the aseptic connector assembly, and the aseptic connector assembly includes one of a triclamp aseptic connector or a hose assembly. Single use feed containers include an aseptic connector assembly.

FIELD OF DISCLOSURE

The present disclosure generally relates to large scale bioreactor systems and, more particularly, to integration of perfusion devices and/or single use feed containers with large scale bioreactors and associated management of pressure.

BACKGROUND

The use of perfusion in a cell culture bioreactor allows improved performance compared to a traditional fed batch cell culture process. Perfusion allows continuous addition of nutrients to the cell culture and continuous removal of metabolic byproducts. In fed batch mode, the tank volume limits the cell culture process, and all byproducts are contained inside the bioreactor until a harvest.

Traditionally, perfusion has been applied to small scale bioreactors, for example, up to 2000 L in working volume. Pressure realized by the perfusion device is greater on a large scale bioreactor due to an increased height of liquid (static head) above the perfusion device when installed in a traditional location near a bottom of a side of the large scale stainless steel bioreactor. In addition, perfusion devices that contain single use components are particularly sensitive to pressure due to the low pressure rating of the single use components.

SUMMARY

In accordance with a first aspect, a large scale bioreactor system comprises a stainless steel large scale bioreactor having at least one valve assembly, and an aseptic connector assembly coupled to the least one valve assembly of the bioreactor. A perfusion device including an Alternating Tangential Filtration (ATF) assembly with an autoclaved valve assembly is coupled to the aseptic connector assembly, and the aseptic connector assembly includes one of a triclamp aseptic connector or a hose assembly.

In accordance with a second aspect, a large scale bioreactor system comprises a stainless steel large scale bioreactor having a side, and an autoclaved valve assembly coupled to the side of the bioreactor. At least one aseptic connector is coupled to the autoclaved valve assembly, and an irradiated single use perfusion device is coupled to the at least one aseptic connector.

In accordance with yet another aspect, a large scale bioreactor system comprises 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. A plurality of single use perfusion devices are connected to the single use adapter assembly, enabling multiple perfusion units to be coupled to the bioreactor without having to steam-in-place the bioreactor upon coupling multiple single use perfusion units.

In accordance with yet another aspect, a large scale bioreactor system comprises a stainless steel large scale bioreactor, and at least one stainless steel transfer panel having a plurality of inputs coupled to the bioreactor. A plurality of single use feed containers are coupled to the at least one stainless steel transfer panel at a working level of the bioreactor. Another aspect is that the stainless steel large scale bioreactor may be installed in a pit depression in a floor to facilitate easy access to a probe belt for operations. Another aspect is that the fully-closed stainless steel large scale bioreactor may be installed in an uncontrolled space and operate exclusively at the probe belt and a local addition panel. So configured, the size of the controlled space is reduced, leading to lower operating cost for a manufacturing facility housing the bioreactor.

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

In some embodiments of any aspect, the stainless steel large scale bioreactor is configured to hold a volume of greater than 2,000 L, such as, e.g., a volume in the range of greater than 2,000 L to 20,000 L, such as, e.g., a volume in the range of 10,000 L to 20,000 L.

DETAILED DESCRIPTION

Generally, a large scale bioreactor system is disclosed. The large scale bioreactor system comprises a stainless steel large scale bioreactor having at least one valve assembly, an aseptic connector assembly coupled to the at least one valve assembly of the bioreactor, and a single use or reusable perfusion device coupled to the aseptic connector assembly. The single use perfusion device includes an perfusion filter assembly with an autoclaved valve assembly coupled to the aseptic connector assembly. So configured, a new perfusion device may be installed while the stainless steel large scale bioreactor is running a cell culture by repeating a steam-in-place of the aseptic connector assembly, or through the use of an aseptic connector valve assembly.

Referring now toFIGS.1-5, schematic representations of a large scale bioreactor system10(FIG.4) according to several aspects of the present disclosure are depicted. As depicted inFIG.1, the large scale bioreactor system10includes a stainless steel large scale bioreactor12having at least one valve assembly14. In these examples, the stainless steel large scale bioreactor12is a steam-in-place SIP bioreactor BRX with large scale capacity. For example, the large scale bioreactor12is configured to hold a volume of fluid of up to 10,000 L or more in one example. The at least one valve assembly14of the steam-in-place large scale bioreactor12enables the steam-in-place large scale bioreactor12to be completely sterilized before perfusion begins. Specifically, the at least one valve assembly14includes a first valve16, a second valve18, a third valve20disposed downstream from each of the first and second valves16,18, and a fourth valve21. Steam, such as clean steam CS, flows into the third valve20, upwardly into the first and second valves16,18, and to a port24. Additionally, after flowing into the third valve20, the steam also flows downwardly into the fourth valve21and through a steam trap22in which clean steam condensate CSC is released, for example, to sterilize the bioreactor12and create a steam sterilized aseptic environment. In one example, when a SIP cycle is finished, condensate, such as clean steam condensate CSC, drains from the steam trap22. In addition, process waste PW flows out of the third and fourth valves20,21during cleaning, emptying into a drain, as is understood by those having ordinary skill. After use, the steam-in-place bioreactor12is cleaned in-place and the steam-in-place process of sterilization occurs again before any further use. The at least one valve assembly14also includes the port24for coupling to an aseptic connector valve assembly26, such as an autoclaved aseptic connector valve assembly26, as depicted inFIG.2, or another device, as explained more below.

Referring now toFIG.2, the aseptic connector valve assembly26is coupled to the at least one valve assembly14. Specifically, and in this example, the aseptic connector valve assembly26is coupled to the port24of the at least one valve assembly14and includes a first valve28and a second valve30disposed downstream from the first valve28. A steam trap32is disposed further downstream from the second valve30. In this example, the autoclaved aseptic connector valve assembly26also includes a triclamp aseptic connector34coupled to the first valve28. The aseptic connector valve assembly26enables sterilized connectivity of a single use perfusion device or a reusable perfusion device to the large scale bioreactor12. The triclamp aseptic connector34is configured to couple to a perfusion device, as explained more below. As will be understood, and in one example, the triclamp aseptic connector34, and the first and second valves28,30may be preassembled and autoclaved before connecting to the at least one valve assembly14for steam sterilization, for example.

Referring now toFIG.3, the large scale bioreactor12having the at least one valve assembly14and the aseptic connector valve assembly26coupled to the at least one valve assembly14is again depicted. In this example, however, the aseptic connector valve assembly26is sterilized independent of the steam-in-place large scale bioreactor12. Specifically, if during a cell culture run in the large scale bioreactor12, it is determined that a perfusion device needs to be serviced and/or replaced, for example, the run of the large scale bioreactor12may continue. This is because the aseptic connector valve assembly26is able to steam the first valve28and the second valve30, for example, and all other parts of the aseptic connector valve assembly26to provide a sterilized connector assembly for coupling to the repaired and/or new perfusion device.

Referring now toFIG.4, a schematic representation of the large scale bioreactor system10according to an aspect of the present disclosure is depicted. More specifically, the stainless steel large scale bioreactor12having the at least one valve assembly14and the aseptic connector valve assembly26coupled to the at least one valve assembly14is depicted. In addition, a perfusion device36, which may be a reusable perfusion device or a single use perfusion device, is coupled to the triclamp aseptic connector34of the aseptic connector valve assembly26. As depicted in bothFIG.4and the close up view of the perfusion device36inFIG.5, the perfusion device36includes an Alternating Tangential Filtration (ATF) assembly38with an autoclaved valve assembly40operatively coupled to the aseptic connector34of the aseptic connector valve assembly26. So configured, the large scale bioreactor system10enables connectivity of the perfusion device36, such as a reusable perfusion device or a single use perfusion device, to the stainless steel large scale bioreactor12without additional steam sterilization.

As also depicted inFIG.5, the autoclaved valve assembly40of the perfusion device36includes a first valve42and a second valve44disposed downstream from the first valve42. In this example, the autoclaved valve assembly40is cleaned out of place (COP) and assembled prior to autoclaving in preparation for connection to the steam-in-place bioreactor12. Generally, COP denotes systems and equipment, such as the autoclaved valve assembly40in this example, which are one or more of disassembled, relocated, or specialty treated to clean and sanitize. Said another way, COP is defined as a method of cleaning equipment items by removing them from their operational area and taking them to a designated cleaning station for cleaning. In addition, an aseptic connector46is operatively coupled to the first valve42and directly coupled to the triclamp aseptic connector34of the aseptic connector assembly26, as depicted inFIG.4. The aseptic connector assembly26allows connectivity between the single use perfusion device36and the stainless steel large scale bioreactor12and provides an ability to restream on subsequent perfusion devices during a single run of the bioreactor12. Said another way, with this configuration, the perfusion device36and/or a new perfusion device may be installed while the stainless steel large scale bioreactor12is running a cell culture, by repeating the steam-in-place or by using the aseptic connector assembly26, for example.

Referring now toFIG.6, another large scale bioreactor system100is depicted. Like the large scale bioreactor system10ofFIGS.1-5, the large scale bioreactor system also includes a stainless steel large scale bioreactor112having a side113and a first valve assembly114, such as an autoclaved valve assembly114, coupled to the side113of the stainless steel large scale bioreactor112. The autoclaved valve assembly114includes a first valve116, a second valve118downstream from the first valve116, a third valve120downstream from both the first and second valves116,118, and a fourth valve121downstream from the third valve120. Again like the valve assembly14of the system10ofFIGS.1-5, steam, such as clean steam CS, flows into the third valve120, upwardly to the first and second valves116,118and to the port134. Additionally, after flowing into the third valve120, steam flows downwardly through the fourth valve121and through a steam trap122to sterilize the bioreactor112and create a steam-in-place process of sterilization before further use, for example. In this large scale bioreactor system100, however, there is only at least one aseptic connector134coupled to the autoclaved valve assembly114, such as the first valve116of the autoclaved valve assembly114, and not a second valve assembly, like the aseptic connector assembly26of the system10ofFIGS.1-5. This same autoclaved valve assembly114enables connection via the at least one aseptic connector134of the autoclaved valve assembly114to a factory assembled and irradiated perfusion device136, which may be a single use perfusion device or a reusable perfusion device, for example. More specifically, the irradiated perfusion device136includes a first aseptic connector138that is coupled to the at least one aseptic connector134, as depicted inFIG.6. The irradiated perfusion device136also includes a second aseptic connector140that is 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 the side113of the large scale stainless steel bioreactor112on a different valve assembly analogous to the valve assembly114. As a result, multiple perfusion devices are able to be aseptically coupled to the large scale stainless steel bioreactor112without additional steam sterilization.

Referring now toFIG.7, a schematic view of another large scale bioreactor system200of the present disclosure is depicted. Like the large scale bioreactor system10ofFIG.4, for example, the large scale bioreactor system200includes a stainless steel large scale bioreactor212having a side wall213and at least one valve assembly214disposed on the side wall213. Again like the large scale bioreactor system10ofFIG.4, the same valve assembly214of the large scale bioreactor200ofFIG.7enables connection of an autoclaved perfusion device and cleaning-in-place of the valve assembly214, but without the use of an aseptic connector, such as the aseptic connector34ofFIG.4. Instead, a hose assembly is used as an alternate connector to couple the stainless steel large scale bioreactor212to the autoclaved perfusion device, as explained more below.

More specifically, and as depicted inFIG.7, the stainless steel large scale bioreactor212of the large scale bioreactor system200is also a steam-in-place bioreactor with large scale capacity. For example, the large scale bioreactor212is configured to hold a volume of fluid of up to or greater than 10,000 L in one example. The at least one valve assembly214of the steam-in-place large scale bioreactor212enables the steam-in-place, stainless steel large scale bioreactor212to be completely sterilized when perfusion begins. Specifically, and like the at least one valve assembly14of the bioreactor system10ofFIG.4, the at least one valve assembly214includes a first valve216, a second valve218, and a third valve220disposed downstream from each of the first and second valves216,218. A fourth valve221may also be disposed downstream from the third valve220, as depicted. Steam, such as clean steam CS, again flows into the third valve220, upwardly to the first and second valves116,118and to a port224. Additionally, after flowing into the third valve220, steam also flows downwardly into the fourth valve221and through a steam trap222to sterilize the bioreactor212and create a steam sterilized aseptic environment. In addition, process waste PW again may flow out of the third and fourth valves220,221during cleaning, emptying into a drain, as is understood by those having ordinary skill. After use, the steam-in-place bioreactor212is cleaned in-place and the steam-in-place process of sterilization occurs again before any further use. The at least one valve assembly214also includes the port224for coupling to a hose assembly235, which in turn couples to a perfusion device or another device, as explained more below.

The hose assembly235includes a hose body237having a first end237A and a second end237B. The first end237A is removably coupled to the port224of the at least one valve assembly214and the second end237B is coupled to an autoclaved perfusion device236, which is functionally equivalent to the perfusion device36of the system10ofFIG.4, for example, but without a single-use connector, such as the single-use connector46ofFIG.5. The autoclaved perfusion device236may include a single-use perfusion device or a reusable perfusion device and still fall within the scope of the present disclosure. The autoclaved perfusion device236includes an ATF assembly238with an autoclaved valve assembly240coupled to the second end237B of the hose body237. The autoclaved valve assembly240of the autoclaved perfusion device236includes a first valve242, and a second valve244disposed downstream from the first valve242. The second end237B of the hose body237is operatively coupled to a connector of the first valve242. The hose assembly235allows connectivity between the autoclaved device236and the stainless steel large scale bioreactor212and provides an ability to restream on subsequent perfusion devices during a single run of the stainless steel large scale bioreactor212.

More specifically, and referring now toFIG.8, the hose assembly235may be removed from connection to the perfusion device236and coupled to another one of multiple ports224on the same large scale bioreactor212. A clean-in-place valve assembly250allows the at least one valve assembly214to be cleaned, which enables another perfusion device, such as a single use perfusion device, to be aseptically coupled to the stainless steel large scale bioreactor212via the hose assembly235. In this example, a clean-in-place steam supply of the clean-in-place valve assembly250upwardly flows into the third valve220and the second valve218of the at least one valve assembly214, up through the first valve216, to port224and through the hose assembly235. In addition, the clean-in-place steam supply also downwardly flows through the fourth valve221downstream from the third valve220. In this way, each of the first, second, third and fourth valves216,218,220and221of the at least one valve assembly214and the port224are cleaned. While this description refers to the at least one valve assembly214, it will be understood that the at least one valve assembly214may include multiple valve assemblies (not shown), and the clean-in-place steam supply of the clean-in-place valve assembly250may also flow through each of the valves and ports of another one of the multiple valve assemblies, allowing multiple valve assemblies to be cleaned.

Referring now toFIG.9, another schematic representation of another large scale bioreactor system300of the present disclosure is depicted. The large scale bioreactor system300includes a stainless steel large scale bioreactor312having a side313and at least one valve assembly314coupled to the side313and an autoclaved valve assembly326coupled to the at least one valve assembly314. In one example, the stainless steel large scale bioreactor312is again a steam-in-place bioreactor with large scale capacity. For example, the large scale bioreactor312is configured to hold a volume of fluid of up to or greater than 10,000 L in one example. The at least one valve assembly314of the steam-in-place large scale bioreactor312enables the steam-in-place large scale bioreactor312to be completely sterilized when perfusion begins. Specifically, and like the at least one valve assembly14ofFIG.4, for example, the at least one valve assembly314includes a first valve316, a second valve318, a third valve320disposed downstream from each of the first and second valves316,318, and a fourth valve321disposed downstream from the third valve320, as depicted. Steam, again such as clean steam CS, flows into the third valve320and upwardly into the second and first valves318,316and a port324to sterilize the at least one valve assembly314and the bioreactor312. In addition, after flowing through the third valve320steam also flows downwardly through the fourth valve321and through a steam trap322to sterilize the bioreactor312and create a steam sterilized aseptic environment. In addition, process waste PW flows out of the fourth valve321during cleaning, emptying into a drain, as is understood by those having ordinary skill. After use, the steam-in-place bioreactor312is cleaned in-place and the steam-in-place process of sterilization occurs again before any further use. The at least one valve assembly314again also includes the port324for coupling to the autoclaved valve assembly326and an aseptic connector334.

Unlike the other bioreactor systems10,100, and200, the bioreactor system300includes an adapter assembly360, such as a single use adapter assembly and/or a wye assembly, which is configured to be and/or is coupled to the aseptic connector334, as also depicted inFIG.9. So configured, the wye assembly360is coupled to the at least one valve assembly314via the aseptic connector334in this example.

The adapter assembly360, such as the wye assembly360, includes a connector361that is directly coupled to the aseptic connector334(which is ultimately coupled to the port324), a pair of tubes362outwardly extending from the connector361, a first aseptic connector364coupled to one of the tubes362, and a second aseptic connector366coupled to the other tube362. Each of the first aseptic connector364and the second aseptic connector366of the wye assembly360is configured to be coupled to a reusable or single use perfusion device, such as any one of the foregoing perfusion devices36,136, and236. This enables multiple perfusion devices to be operatively coupled to the stainless steel large scale bioreactor312via the port324of the valve assembly314, for example, without having to steam-in-place the stainless steel large scale bioreactor312upon coupling multiple reusable perfusion devices or single use perfusion devices. In another example, the adapter assembly360, such as the wye assembly360, is a first wye assembly360and a second wye assembly (not shown) may be coupled to the first wye assembly. So configured, coupling one or more additional wye assemblies to the first wye assembly360enables more than two reusable or single use perfusion devices and multiple reusable or single use perfusion devices to be operatively coupled to the stainless steel large scale bioreactor312, again without having to steam-in-place the stainless steel large scale bioreactor312upon coupling the multiple single use perfusion devices. Likewise, a greater number of perfusion devices can be connected to the large scale bioreactor312and each device does not require a distinct reactor port.

Referring now toFIG.10, in another example, the large scale bioreactor system300is depicted and includes the large scale stainless steel bioreactor312that is configured to hold a volume of fluid of up to or greater than 10,000 L in one example. In this example, the at least one valve assembly of the large scale bioreactor system300includes the at least one valve assembly380, which is configured to be coupled to a transfer panel (not depicted inFIG.10), mounted locally or at a remote distance from the large scale stainless steel bioreactor312for the purpose of connecting and transferring feed liquids held in single use containers to the large scale stainless steel bioreactor312. The valve assembly380is configured to enable steam sterilization of a transfer line385and connection to a single use feed container. Steam from the large scale bioreactor300passes out through at least one transfer line, such as the transfer line385in one example, and to a triclamp port324and the aseptic connector assembly334, and through a first valve381, a second valve382, a third valve383, and a fourth valve384and downwardly to the steam trap322. As such, the entire line is rendered sterile up to the aseptic connector334. Likewise, cleaning of the line is accomplished by supplying clean-in-place (CIP) solution through the first and second valves381,382and through transfer line385to the large scale bioreactor300. As depicted inFIG.10, the ports324of the first and second valves381,382are coupled to a single use adapter, such as the aseptic connector334, which are designed to tolerate direct exposure to clean steam at sterilizing temperatures and pressures. As further depicted, the single use adapter, such as the aseptic connector assembly334, in turn is coupled to a single use manifold370. In this example, each single use manifold370includes eight or more arms371having inlets372, each of which is configured to be coupled to a feed container373, such as a single use feed container. In another example, the single use manifold370may include more or fewer than eight arms371, such as four or six arms and still fall within the scope of the present disclosure. So configured, multiple feed containers are able to be operatively coupled to the stainless steel large scale bioreactor312without additional steaming and cleaning. One or more additional feed containers373can be connected using aseptic connectors372while the large scale stainless steel bioreactor312is in operation. In addition, one or more additional single use manifolds370may be connected to inlets372via the aseptic connector(s) on the first single use manifold assembly370to further increase the number of feed containers373that may be connected.

Referring again toFIG.10, in the event that an aseptic connector is to be employed that cannot tolerate direct exposure to clean steam at sterilizing temperatures and pressures, an alternate configuration enables isolation of the aseptic connector from the steam. For example, one or more valve assemblies390A and390B may be prepared by coupling a valve or two valves, such as valve391A of the valve assembly390A, and first and second valves391B,393B of the valve assembly390B, which may be any valve with a low hold up volume on the side to the steamed to an aseptic connector, such as an aseptic connector392A or an aseptic connector392B, and autoclaved. Each of valve assembly390A and valve assembly390B may be inserted in a valve assembly that may be installed on a local or remote transfer panel, such as at least one valve assembly380A depicted inFIG.10, by coupling with triclamp connectors, for example. From that point forward, the cleaning, steaming and operation are the same as that described above relative toFIG.9. By using a single use manifold assembly with aseptic connector392, a larger inner diameter, lower shear and higher flow rates are afforded than can be achieved using an aseptic connector that can be directly exposed to clean steam at sterilization temperatures and pressures. However, since the main transfer line385and the at least one valve assembly380,380A are the same, either type of aseptic connector may be used and a similar plurality of feed containers373can be connected. Alternatively, a single connection to the feed container373may be used when higher flow rates or lower shear are required.

Referring now toFIG.11, the bioreactor system100ofFIG.6is again depicted and includes all of the same features of the bioreactor system100except for the location of the valve assembly114relative to the stainless steel large scale bioreactor112. Specifically, the valve assembly114is mounted higher on the side113of the stainless steel large scale bioreactor112, which in turns results in the perfusion device136also being mounted higher on the side113of the stainless steel large scale bioreactor112. In particular, in this example, the side113of the bioreactor112includes a bottom end113A and a top end113B, and the valve assembly114is coupled to the side113of the bioreactor in a location closer to the top end113B of the side113than the bottom end113A, as depicted inFIG.11. So configured, the height of liquid L disposed above the perfusion device136, and therefore pressure, is reduced.

Generally, pressure realized by the single use perfusion device, such as the perfusion device36,136,236, is greater on the stainless steel large scale bioreactor12,112,212,312, due to the increased height of liquid (static head) above the perfusion device36,136,236when installed in a typical location near a bottom side, such as the bottom end113A, of the stainless steel large scale bioreactor12,112,212,312. Perfusion devices that include single use components are particularly sensitive to greater pressure due to the low pressure rating of the single use components, for example. Thus, by mounting the valve assembly114and, thus, the perfusion device136ofFIG.11, near the top end113B of the bioreactor112, pressure is reduced.

Still referring toFIG.11, the perfusion device136, such as the single use perfusion device, includes at least one pressure sensor180, and may include additional pressure sensors. In this example, the perfusion device136, such as an RFT single use perfusion device, includes a first pressure sensor180, a second pressure sensor182, and a third pressure sensor184. In addition, the perfusion device136also includes a circulation pump186disposed upstream from the first pressure sensor180, and a permeate pump188disposed downstream from the third sensor184.

As further depicted inFIG.11, a control system190for monitoring pressure of at least one perfusion device136is communicatively coupled via a network, such as a wireless network192or a wired connection193, to the single use perfusion device136. While the control system190is operatively coupled to the perfusion device136, the control system190may also optionally be coupled to any one of the other foregoing described perfusion devices36,236and still fall within the scope of the present disclosure. In one example, the control system190includes a computing device194having a memory195, a processor196, and a network interface197. It will be understood that the computing device194may also include any known input, receiver and transmitter. In addition, the control system190further includes an alarm198. So configured, the control system190monitors the pressure at one or more of the first, second and third pressure sensors180,182,184. If the pressure approaches a pre-defined safe limit, the alarm198will be activated to notify users so the bioreactor system100may be investigated and evaluated. If the pressure being monitored by the control system190approaches still closer to the pre-defined safe limit, the control system190will automatically reduce the flow rate of fluid, and thus the pressure, to maintain a safe operating pressure for the bioreactor system100. If the pressure being monitored by the control system190achieves the safe pressure limit, the control system190will stop the circulating pump186, preventing damage to the perfusion device136.

Referring now toFIG.12, any one of the foregoing large scale stainless steel bioreactors12,112,212,312may be disposed in a pit402, as depicted inFIG.12. In addition, the valve assembly14,114,214,314is then mounted near the bottom end13A,113A,213A,313A of the side13,113,213,313of the large scale stainless steel bioreactor12,112,212,312and against a bottom weld seam404of a body of the bioreactor12,112,212,312. This enables the single use perfusion device36,136,236to be positioned higher, minimizing hold-up volume between the perfusion device36,136,236and the large scale stainless steel bioreactor12,112,212,312. Alternatively, by adjusting the depth of the pit, the pit402may be used to locate any one of the perfusion devices36,136,236higher on the side13,113,213of the bioreactor12,112,212to also reduce pressure.

Referring now toFIG.13, another schematic diagram of another large scale bioreactor system500of the present disclosure is depicted. The large scale bioreactor system500includes a stainless steel large scale bioreactor512having a side513with a bottom end513A and a top end513B. In addition, at least one stainless steel transfer panel520having a plurality of inputs522is coupled to the side513of the stainless steel large scale bioreactor512at the top end513B of the side513. Further, a plurality of single use feed containers524is coupled to one or more inputs522of the plurality of inputs522of the transfer panel520at a working level526of the stainless steel large scale bioreactor512. While one stainless steel transfer panel520is depicted inFIG.13, it will be appreciated that multiple stainless steel transfer panels may alternatively and/or additionally be coupled to the bioreactor512and still fall within the scope of the present disclosure. So configured, this prevents a user from having to climb a stairwell every time they connect something to the bioreactor512. In addition, it allows making connections to single use feed containers inside a smaller clean space without locating the entire bioreactor512inside the clean space, for example, with some portion of the bioreactor located in a controlled, non-classified (CNC) space. This also enables users to work from a clean controlled space, operating the bioreactor512from the clean controlled space without having to access the top end of the bioreactor512.

In addition, it will be appreciated that all components of any one or more of the foregoing bioreactor systems10,100,200,300and500needing steaming may be steamed while the associated bioreactor12,112,212,312,512is being steamed. In this scenario, the steam source is from the bioreactor tank12,112,312,412,512and steam flows out through the valves and to the steam traps depicted in the figures. For example, in the bioreactor system10ofFIG.2, steam may flow out of the bioreactor12through the first and second valves16,18to the third and fourth valves20,21and to the steam trap22. Likewise, in the bioreactor system100ofFIG.6, steam may also flow out of the bioreactor112, through the first, second, third, and fourth valves116,118,120,121and to the steam trap122. In a similar manner, in the bioreactor system200ofFIG.9, steam may again flow out of the bioreactor212into the first, second, third and fourth valves216,218,220,221and to the steam trap222. Similarly, in the bioreactor system300, the steam may again flow from the bioreactor312and into the first and second valves316,318, through the third and fourth valves320,321, and to the stream trap322.

In view of the foregoing, it will be appreciated that any one or more the foregoing bioreactors12,112,212,312may be integrated with any one or more of the foregoing perfusion devices, such as the reusable perfusion devices or single use perfusion devices36,136,236according to one or more of the following methods. Specifically, and according to one example, a method of integrating at least one single use perfusion device36,136,236with a stainless steel large scale bioreactor12,112,212,312comprises coupling one of: (1) a connector assembly26, such as an aseptic connector assembly26to at least one valve assembly14,214of the stainless steel large scale bioreactor12,112,212,312; or (2) an autoclaved valve assembly116to a side113of the stainless steel large scale bioreactor112. In addition, the method further comprises coupling one of: (1) an autoclaved valve assembly40,240of an ATF assembly of a single use perfusion device36,236to the connector assembly26; or (2) an irradiated perfusion device136to the autoclaved valve assembly. Moreover, the method still further comprises managing pressure of the perfusion device36,136,236via at least one pressure sensor of the perfusion device36,136,236and automatically reducing one or more of a flow rate or a pressure in the perfusion device36,136,236by a control system190upon detecting a pressure greater than a safe limit pressure.

In one example, the method comprises coupling the aseptic connector assembly to the at least one valve assembly14,214of the stainless steel large scale bioreactor12,212and coupling the autoclaved valve assembly40to the aseptic connector assembly26wherein the aseptic connector assembly26, includes one of the triclamp connector assembly or a hose assembly. In another example, the method comprises coupling the autoclaved valve assembly114to the side113of the stainless steel large scale bioreactor112and coupling the perfusion device136to one of a plurality of the autoclaved valve assemblies114. While only one autoclaved valve assembly114is depicted, it will be understood that two or more autoclaved valve assemblies114may be coupled to the perfusion device136and still fall within the scope of the present disclosure.

In yet another example, the method comprises coupling a connector assembly360to at least one valve assembly314of a stainless steel large scale bioreactor312, the connector assembly360including a wye connector assembly. In addition, the method further comprises coupling the autoclaved valve assembly40of an ATF assembly of at least one perfusion device36to the connector assembly360, wherein the at least one perfusion device36or feed containers373,524comprise a plurality of perfusion devices or feed containers connected to the wye connector assembly. This enables multiple perfusion devices or feed containers to be coupled to the bioreactor12,112,212,312without having to steam-in-place the bioreactor12,112,212,312upon coupling the multiple perfusion devices or feed containers.

The above description describes various bioreactor systems and methods of integrating at least one single use perfusion device and/or at least one feed container with a stainless steel large scale bioreactor. It will be appreciated the systems and methods of the present disclosure include several advantages. For example, the systems and methods described enable connection of perfusion devices to large scale bioreactors (e.g., bioreactors with a capacity of greater than 2,000 L), replacement of a perfusion device or a feed container during a cell culture run, and pressure management for perfusion devices, all while creating a steam sterilized aseptic environment. In addition, the same valve assembly at the bioreactor enables both connection of a factory assembled and irradiated single use perfusion device and an autoclaved perfusion device and cleaning-in-place of the valve assembly of the bioreactor, without the use of an aseptic connector, for example.

Although the foregoing systems and methods, and elements thereof, have been described in terms of exemplary embodiments, they are not limited thereto. The detailed description is to be construed as exemplary only and does not describe every possible embodiment of the invention because describing every possible embodiment would be impractical, if not impossible. Numerous alternative embodiments could 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 defining the invention.

It should be understood that the legal scope of the invention is defined by the words of the claims set forth at the end of this patent. The appended claims should be construed broadly to include other variants and embodiments of same, which may be made by those skilled in the art without departing from the scope and range of equivalents of the devices, systems, methods, and their elements.