Flexible facility configuration for the manufacture of therapeutic products

Abstract

Disclosed herein is a flexible facility for manufacturing at least one therapeutic product, the flexible facility comprising a reconfigurable single-use area and a stainless steel area, which may optionally be partitioned by partitions.

Description

【Technical Field】 【0001】 Cross - reference to Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 356,361, filed on June 28, 2022, the entire content of which is incorporated herein by reference. 【0002】 The present disclosure relates to a flexible manufacturing facility that enables the production of therapeutic products, such as active pharmaceutical ingredients, cell therapies, etc., which may require different equipment, such as single - use systems and stainless - steel equipment, in sequential or parallel campaigns. In some embodiments, the flexible manufacturing facility combines a reconfigurable single - use area and a conventional stainless - steel area, and the reconfigurable single - use area and the stainless - steel area are optionally partitioned by partitions. 【Background Art】 【0003】 Biopharmaceuticals are used worldwide in a variety of therapeutic areas ranging from oncology to cardiology and beyond. However, the costs associated with the manufacture of biopharmaceuticals are high due to their complex production methods and the concentration on single modalities and / or single products in typical biomanufacturing facilities. In the manufacture of biopharmaceuticals, a multi-step process is typically used that includes scale-up of cell culture, cell culture for large-scale production, and purification of the desired biopharmaceutical after cell harvest, and these steps are often specific to a given modality or product. The complexity of manufacturing has been increasing in recent years due to the availability of many new modalities such as monoclonal antibodies, BiTE® molecules, fusion proteins, CAR-T cell therapies, RNA therapies, etc. Although improvements in all aspects of production have reduced the manufacturing costs on a commercial scale for many biopharmaceuticals, the initial costs associated with the construction and maintenance of biomanufacturing facilities, including the initial costs associated with campaign switching to change or expand the modalities and / or the types of products manufactured in a given facility, can still be prohibitively high. Therefore, there is a need in the art for new, reconfigurable biomanufacturing facility configurations that enable the manufacture of multiple products and / or modalities in a single facility over its entire useful life, at reduced cost and reduced time, and that facilitate long-term supply planning. 【0004】 Biopharmaceuticals are generally produced in an industry-standard platform called "fed-batch". In the fed-batch process, a semi-open system is used in which one or more nutrients are supplied in bolus to the bioreactor during cell culture. The fed-batch production process is often carried out in large stainless-steel bioreactors, and much of the cost associated with biopharmaceutical manufacturing can be attributed to the capital investment required to build and maintain the infrastructure necessary to support the production and purification of a particular biopharmaceutical. Exemplarily, the construction of a fed-batch facility often requires a capital investment of approximately $1.5 billion and approximately six years to obtain approval. Furthermore, in conventional configurations for manufacturing biological products such as monoclonal antibodies and microbial products, efficient use of facility space is not possible, for example, due to the need for large stainless-steel tanks and extensive stainless-steel fixed piping to support the fed-batch process. Additionally, the layout of conventional biomanufacturing facilities, which often includes extensive fixed piping and large equipment, may limit its ability to be easily reconfigured or expanded to manufacture new product lines. This limitation is product-specific and is often exacerbated by the diverse platforms used to manufacture biopharmaceuticals, which lack compatibility. 【0005】 An ideal biomanufacturing facility should be inexpensive to construct, quickly expandable and reconfigurable to rapidly and cost-effectively incorporate new products and processes. Additionally, it would be advantageous for such a manufacturing facility to be able to maintain and improve the high levels of quality required in current Good Manufacturing Practice (cGMP) for drug manufacturing (e.g., compliance with 22 C.F.R. Part 11). 【0006】 Single-use production technologies that simplify equipment and facility design compared to reusable systems that require cleaning and sterilization between production batches are becoming increasingly prevalent in biomanufacturing facilities. Single-use components do not require cleaning after each batch and are a cost-effective means of manufacturing biopharmaceuticals. However, the manufacturing requirements for certain biopharmaceuticals exceed the current capacity or other capabilities achievable with single-use technology. Therefore, for certain biopharmaceuticals with high volume requirements, it is not possible to avoid reusable large-capacity stainless steel containers. Such containers must be connected by stainless steel piping to other unit operation systems, water supply systems, media and buffer supply systems, and in-place cleaning and in-place steam sterilization systems. As noted above, the fabrication and installation of these stainless steel containers and all the utilities that support them are expensive and require a significant lead time for design and manufacture. 【0007】 Accordingly, there is a need for a flexible manufacturing facility that enables a user to efficiently allocate common resources between product lines and that can be easily reconfigured to accommodate new manufacturing processes or new manufacturing lines, with a reduction in facility downtime. SUMMARY OF THE INVENTION MEANS FOR SOLVING THE PROBLEM 【0008】 The present disclosure provides a flexible manufacturing facility for the production of biopharmaceuticals, the flexible manufacturing facility being adaptable to the specific needs of one or more biopharmaceuticals, such as one or more biopharmaceuticals of different modalities. This novel facility layout utilizes a reconfigurable single-use area and a stainless steel area, which may optionally be partitioned by partitions. 【0009】 Disclosed herein is a flexible facility for manufacturing at least one therapeutic product, A reconfigurable single-use area comprising at least one single-use bioreactor, wherein one or more of the at least one single-use bioreactor has a capacity of at least 1000 L (e.g., at least 2000 L, more than 2000 L), the reconfigurable single-use area, A stainless steel area comprising at least one stainless steel bioreactor, wherein each stainless steel bioreactor in the stainless steel area has a capacity greater than 5000 L, the stainless steel area, Comprising, One or more of the at least one stainless steel bioreactor is fluidly connected to one or more of the at least one single-use bioreactor, Flexible facility. 【0010】 In some embodiments, one or more of the at least one single-use bioreactor has a capacity of 1000 L. In some embodiments, a plurality of the at least one single-use bioreactor has a capacity of 1000 L. 【0011】 In some embodiments, one or more of the at least one single-use bioreactor has a capacity of at least 2000 L. In some embodiments, one or more of the at least one single-use bioreactor has a capacity of 2000 L. In some embodiments, a plurality of the at least one single-use bioreactor has a capacity of 2000 L. 【0012】 In some embodiments, one or more of the at least one single-use bioreactor has a capacity of at least 3000 L. In some embodiments, one or more of the at least one single-use bioreactor has a capacity of 3000 L. In some embodiments, a plurality of the at least one single-use bioreactor has a capacity of 3000 L. 【0013】 In some embodiments, one or more of the at least one single-use bioreactor has a capacity of at least 5000 L. In some embodiments, one or more of the at least one single-use bioreactor has a capacity of 5000 L. In some embodiments, a plurality of the at least one single-use bioreactor has a capacity of 5000 L. 【0014】 In some embodiments, one or more of the at least one single-use bioreactor has a capacity of 1000 L, 2000 L, 3000 L, or 5000 L. In some embodiments, a plurality of the at least one single-use bioreactor has a capacity independently selected from 1000 L, 2000 L, 3000 L, and 5000 L. 【0015】 In some embodiments, one or more of the at least one single-use bioreactor has a capacity of 200 L. In some embodiments, one or more of the at least one single-use bioreactor has a capacity of 500 L. 【0016】 Non-limiting examples of equipment that can be used in a flexible facility include bioreactors, disk stack centrifuges, single-use centrifuges, tangential flow filtration (TFF) skids, depth filtration skids, in-line dilution skids, chromatography columns with associated control equipment, media tanks, harvest tanks, purification vessels, depth filter holders, water softening and / or dechlorination systems, clean steam generators, water for injection (WFI) storage tanks, WFI break tanks, WFI stills, cooling towers, switchboards, emergency generators, chillers, pumps, autoclaves, air handling units, process waste neutralization (e.g., fiber reinforced plastic (FRP), etc.) equipment, bio-waste collection and / or inactivation systems, stationary cleaning systems, stationary steam sterilization systems, parts washers, and / or other equipment. 【0017】 As non-limiting examples, but not by way of limitation, U.S. Patent Application Publication No. 2013 / 0280797, U.S. Patent Application Publication No. 2012 / 0077429, U.S. Patent Application Publication No. 2011 / 0280797, and U.S. Patent Application Publication No. 2009 / 0305626, and U.S. Patent No. 8,298,054, U.S. Patent No. 7,629,167, and U.S. Patent No. 5,656,491 (which are incorporated herein by reference) describe exemplary facility areas, equipment, and / or systems having features that may be suitable for use in the flexible facilities described herein. 【0018】 In some embodiments, the stainless steel area, considered as a whole, has a lower air classification than the reconfigurable single-use area, considered as a whole. In some embodiments, the stainless steel area, considered as a whole, has a lower architectural finish than the reconfigurable single-use area, considered as a whole. In some embodiments, the stainless steel area, considered as a whole, has a lower air classification and a lower architectural finish than the reconfigurable single-use area, considered as a whole. 【0019】 In some embodiments, the stainless steel area is an unclassified manufacturing space. In some embodiments, at least a portion (e.g., the seed train area, the purification area) of the reconfigurable single-use area is a classified manufacturing space. The integrity and environmental control inside at least a portion of the reconfigurable single-use area facilitate the use of the unclassified manufacturing space. 【0020】 In some embodiments, one or more of at least one stainless steel bioreactor are supported on the ground. 【0021】 In some embodiments, one or more of at least one stainless steel bioreactor can be suspended from the structure itself. 【0022】 In some embodiments, the stainless - steel area includes a plurality of stainless - steel bioreactors, each having a capacity of at least 10,000 L. 【0023】 In some embodiments, each stainless - steel bioreactor within the stainless - steel area has a capacity of 5000 L to 20,000 L. In some embodiments, each stainless - steel bioreactor within the stainless - steel area has a capacity of 5000 L to 15,000 L. In some embodiments, each stainless - steel bioreactor within the stainless - steel area has a capacity of 5000 L to 10,000 L. 【0024】 In some embodiments, the stainless - steel area does not have a physical barrier separating it from the reconfigurable single - use area. 【0025】 In some embodiments, the reconfigurable single - use area and the stainless - steel area are partitioned by a partition. In some embodiments, the partition is a wall. In some embodiments, the partition is a plastic sheet. 【0026】 In some embodiments, the stainless - steel area is completely enclosed. In some embodiments, the stainless - steel area is completely enclosed by one or more walls and a ceiling. 【0027】 In some embodiments, the stainless - steel area has a separate air handling from the reconfigurable single - use area. 【0028】 In some embodiments, a portion of at least one stainless - steel bioreactor penetrates a wall and is fluidly connected via a pipe to one or more of at least one single - use bioreactor. In some embodiments, this portion is sufficient to facilitate the connection between instrumentation units and / or process units. In some embodiments, this portion is minimally sufficient to facilitate the connection between instrumentation units and / or process units. 【0029】 In some embodiments, at least 1% (e.g., at least 2%, at least 3%, at least 4%, at least 5%; 1%, 2%, 3%, 4%, 5%) of the external container surface area of at least one stainless steel bioreactor penetrates the wall and is fluidly connected via a tube to one or more of at least one single-use bioreactor. In some embodiments, 3% - 7% (e.g., 3.5% - 6.5%, 4% - 6%, 4.5% - 5.5%; 3%, 4%, 5%, 6%, 7%) of the external container surface area of at least one stainless steel bioreactor penetrates the wall and is fluidly connected via a tube to one or more of at least one single-use bioreactor. 【0030】 In some embodiments, the portion penetrating the wall includes one or more components selected from a probe belt for enabling access to bioreactor instrumentation, a perfusion connection, and a connection for feed liquid (e.g., nutrient feed, antifoam-containing liquid). 【0031】 In some embodiments, the fluid connection is configured to enable bypass of the stainless steel area. In some embodiments, the fluid connection is detachable. In some embodiments, the fluid connection can be moved between different bioreactors within a flexible facility. 【0032】 In some embodiments, the stainless - steel area further includes a recovery area. In some embodiments, the recovery area within the stainless - steel area includes one or more centrifuges. In some embodiments, the recovery area within the stainless - steel area includes one or more stainless - steel centrifuges. In some embodiments, the equipment within the stainless - steel area is connected with pipes and / or one or more valves to enable the fluid connections necessary to complete the specific processes being performed. In some embodiments, one or more valves are used to bypass specific equipment not necessary for the process being performed. In some embodiments, one or more valves are used to shut off the fluid flow to enable the replacement of equipment. For example, in some embodiments, an inlet conduit from a seed train is connected to each stainless - steel bioreactor. In some embodiments, an outlet conduit from each stainless - steel bioreactor to a centrifuge for recovery is connected. 【0033】 In some embodiments, the reconfigurable single - use area may include one or more additional spaces independently selected from a cell - culture unit, a pre - viral unit, a post - viral unit, a utility yard, a warehouse, a media buffer facility, an office, a personnel unit, and a production unit. The production unit may be useful for the manufacture of therapeutic products. 【0034】 In some embodiments, the reconfigurable single - use area may include a reagent - preparation area, a seed - train area, and a purification area. 【0035】 In some embodiments, the reconfigurable single-use area further includes equipment for performing downstream purification. In some embodiments, the equipment for performing downstream purification is single-use or stainless steel. In some embodiments, the equipment for performing downstream purification is single-use. In some embodiments, the equipment for performing downstream purification is stainless steel. In some embodiments, some of the equipment for performing downstream purification is stainless steel and some of the equipment for performing downstream purification is single-use. 【0036】 In some embodiments, the stainless steel area further includes equipment for performing downstream purification. In some embodiments, the equipment for performing downstream purification is single-use or stainless steel. In some embodiments, the equipment for performing downstream purification is single-use. In some embodiments, the equipment for performing downstream purification is stainless steel. 【0037】 In some embodiments, the reconfigurable single-use area further includes one or more centrifuges. In some embodiments, the one or more centrifuges are single-use and / or stainless steel equipment. In some embodiments, the one or more centrifuges are single-use equipment. In some embodiments, the one or more centrifuges are stainless steel equipment. 【0038】 In some embodiments, the stainless steel area further includes one or more centrifuges. In some embodiments, the one or more centrifuges are single-use and / or stainless steel equipment. In some embodiments, the one or more centrifuges are single-use equipment. In some embodiments, the one or more centrifuges are stainless steel equipment. 【0039】 In some embodiments, the reconfigurable single-use area includes a reagent preparation area. In some embodiments, the reconfigurable single-use area includes a wall around the reagent preparation area. In some embodiments, the reagent preparation area has separate air handling. In some embodiments, the reconfigurable single-use area includes a seed train area. In some embodiments, the reconfigurable single-use area includes a wall around the seed train area. In some embodiments, the seed train area has separate air handling. In some embodiments, the reconfigurable single-use area includes a purification area. In some embodiments, the reconfigurable single-use area includes a wall around the purification area. In some embodiments, the purification area has separate air handling. 【0040】 In some embodiments, the reconfigurable single-use area includes a reagent preparation area, a seed train area, and a purification area. In some embodiments, the reconfigurable single-use area includes a reagent preparation area, a seed train area, and a purification area, along with one or more partitions that at least partially compartmentalize one or more of the reagent preparation area, the seed train area, and the purification area. In some embodiments, the reconfigurable single-use area includes a reagent preparation area, a seed train area, and a purification area, and the reagent preparation area, the seed train area, and the purification area are not all served by the same air handling system. 【0041】 In some embodiments, the stainless steel area is adjacent to the seed train area, or adjacent to the purification area, or adjacent to both the seed train area and the purification area. 【0042】 In some embodiments, the reconfigurable single-use area includes one or more seed trains. In some embodiments, the reconfigurable single-use area includes one or more wave bioreactors, e.g., one or more wave bioreactors having a capacity of at least 50 L (e.g., 50 L). In some embodiments, the reconfigurable single-use area includes one or more seed bioreactors, such as one or more seed bioreactors having a capacity of at least 500 L (e.g., 500 L) or at least 2,000 L (e.g., 2,000 L). 【0043】 In some embodiments, the reconfigurable single-use area further includes one or more purification skids. In some embodiments, each of the one or more purification skids includes one or more components independently selected from a chromatography skid, a chromatography column, a virus inactivation system, a virus filtration system, an ultrafiltration diafiltration system, and a bulk filtration system. In some embodiments, the reconfigurable single-use area further includes a chromatography skid. In some embodiments, the reconfigurable single-use area further includes a virus inactivation system. In some embodiments, the reconfigurable single-use area further includes a virus filtration system. In some embodiments, the reconfigurable single-use area further includes an ultrafiltration / diafiltration system. In some embodiments, the reconfigurable single-use area further includes a bulk filtration system. In some embodiments, the reconfigurable single-use area further includes a virus inactivation system and a virus filtration system. 【0044】 In some embodiments, one or more purification skids within the flexible facility have separate air handling. 【0045】 In some embodiments, the flexible facility includes a plurality of reconfigurable connections for connecting different equipment (e.g., bioreactors, purification skids, etc.) within the flexible facility. 【0046】 In some embodiments, in the reconfigurable single-use area, utilities are provided overhead, from one or more walls, or from one or more floor-mounted utility distribution units, or a combination of the aforementioned utility distribution options. In some embodiments, in the reconfigurable single-use area, utilities are provided overhead. In some embodiments, the utilities within the reconfigurable single-use area are easily accessible. 【0047】 In some embodiments, multiple devices within the reconfigurable single-use area are movable within the reconfigurable single-use area. 【0048】 In some embodiments, the flexible facility further includes a quality assurance (QA) section, an administrative / office section, plant utilities (e.g., air handling, electricity, and plumbing), a quality control area, a laboratory area, and a receiving area. 【0049】 In some embodiments, the flexible facility further includes one or more additional spaces independently selected from a utility space, a media buffer facility, a dressing room, and a production unit. In some embodiments, the flexible facility further includes a utility space. In some embodiments, the flexible facility further includes a media buffer facility. In some embodiments, the flexible facility further includes a dressing room. In some embodiments, the flexible facility further includes a production unit. In some embodiments, the flexible facility further includes a utility space, a media buffer facility, a dressing room, and a production unit. In some embodiments, at least one additional space (e.g., a utility space, a media buffer facility, a dressing room, and / or a production unit) is located within the reconfigurable single-use area. 【0050】 In some embodiments, the flexible facility includes at least one of a fermentation unit, a pre-virus unit, a post-virus unit, a utility space, a warehouse, a media buffer facility, an office, a personnel unit, and a production unit. 【0051】 In some embodiments, the flexible facility can include any typical manufacturing and cleanroom equipment. 【0052】 In some embodiments, the flexible facility is configured to produce one or more therapeutic products independently selected from monoclonal antibodies, BiTE® molecules, RNA-based drug substances, cell therapy products, and fermentation products. In some embodiments, the flexible facility is configured to produce monoclonal antibodies. In some embodiments, the flexible facility is configured to produce BiTE® molecules. In some embodiments, the flexible facility is configured to produce RNA-based drug substances. In some embodiments, the flexible facility is configured to produce cell therapy products. In some embodiments, the flexible facility is configured to produce fermentation products. 【0053】 In some embodiments, the stainless-steel bioreactor within the stainless-steel area is dedicated to a single therapeutic product. In some embodiments, the entire flexible facility is dedicated to the production of a single therapeutic product. In some embodiments, the stainless-steel bioreactor within the stainless-steel area contains only one cell line for the production of a therapeutic product. 【0054】 In some embodiments, the flexible facility is dedicated to the simultaneous production of more than one (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.) therapeutic products (e.g., simultaneous production of multiple products in, e.g., a seed train, a production bioreactor, and a purification skid). In some embodiments, the flexible facility is dedicated to the simultaneous production of 2 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 3 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 4 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 5 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 6 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 7 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 8 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 9 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 10 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 11 therapeutic products. In some embodiments, the flexible facility is dedicated to the simultaneous production of 12 therapeutic products. 【0055】 In some embodiments, in the flexible facility, during campaign switching, more than one (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.) therapeutic products are produced. For example, in some embodiments, after cell harvest and during purification, the production of a second therapeutic product can be initiated in the seed train. In some embodiments, the stainless steel bioreactor within the stainless steel area contains more than one therapeutic product-producing cell line. 【0056】 In some embodiments, the flexible facility is dedicated to the production of more than one (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, etc.) therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 10 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 15 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 20 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 25 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 30 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 35 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 40 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 45 therapeutic products during its useful life. In some embodiments, the flexible facility is dedicated to the production of up to 50 therapeutic products during its useful life. 【0057】 In some embodiments, the flexible facility is configured to accommodate GMP (Good Manufacturing Practice) processes. In some embodiments, the flexible facility is configured to accommodate GMP processes. In some embodiments, the flexible facility is configured to accommodate non-GMP processes. In some embodiments, the flexible facility is capable of producing GMP clinical products. In some embodiments, the flexible facility is capable of producing GMP commercial products. In some embodiments, the flexible facility is capable of producing non-GMP products. In some embodiments, the flexible facility is capable of producing GMP clinical products and GMP commercial products. In some embodiments, the flexible facility is capable of producing GMP clinical products and non-GMP products. In some embodiments, the flexible facility is capable of producing GMP commercial products and non-GMP products. In some embodiments, the flexible facility is capable of producing GMP clinical products, GMP commercial products, and non-GMP products. 【Brief Description of the Drawings】 【0058】 【Figure 1】 Shows a non-limiting example of a flexible manufacturing facility according to the present disclosure. FIG. 1 is a top view of an exemplary embodiment of a flexible facility. In some embodiments, the flexible facility of FIG. 1 can be constructed in a series of stages such as a first stage, a second stage, and subsequent stages. 【Figure 2A】 Is a schematic diagram showing various components of the flexible manufacturing facility of FIG. 1. 【Figure 2B】 Is a schematic diagram showing some components of at least one stainless steel bioreactor of FIGS. 1 and 2A. 【Figure 2C】 Is a schematic diagram showing at least one purification skid of one or more purification skids of FIGS. 1 and 2A. 【Mode for Carrying Out the Invention】 【0059】 The present disclosure relates to a new manufacturing facility configuration that incorporates state-of-the-art technologies of a next-generation biomanufacturing platform based on single-use equipment and enables process enhancement within a conventional fed-batch facility based on reusable stainless-steel equipment. This new facility configuration can facilitate the simultaneous or sequential production of multiple biopharmaceuticals manufactured using diverse manufacturing platforms. This facility configuration also provides enhanced flexibility with respect to the layout of conventional biomanufacturing facilities. For example, the facility can be configured or reconfigured to accommodate any of the following production technologies, namely, (a) fed-batch cell culture processes (e.g., at scales up to 10,000 L, greater than 10,000 L scale) (e.g., using single-use and / or stainless-steel equipment), (b) single-use cell culture processes at scales up to 2,000 L (e.g., using only single-use technologies), and (c) continuous upstream manufacturing at scales up to 500 L. 【0060】 In addition to providing platform flexibility, the biomanufacturing facilities described herein are expected to exhibit one or more of the following advantages compared to conventional fed-batch facilities with similar production capacity, namely, (a) reduced financial risk due to reduced space requirements, reduced equipment capacity, reduced stainless-steel fixed piping, and reduced automated stainless-steel valves (e.g., capital expenditure less than 50%), (b) more rapid construction with earlier approval acquisition due to improved space utilization, and (c) reduction in carbon emissions and water consumption (e.g., up to 50%) when sustainable building materials are used and dependence on stainless-steel equipment requiring regular cleaning and sterilization procedures is reduced. 【0061】 Referring now to FIGS. 1-2C, a flexible manufacturing facility 10 for manufacturing at least one therapeutic product is shown. The flexible facility 10 includes a reconfigurable single-use area 12 that includes at least one single-use bioreactor 14. In this embodiment, and as further described below, one or more of the at least one single-use bioreactors 14 have a capacity of at least 1000 L. In another example, one or more of the at least one single-use bioreactors 14 have a capacity of at least 2000 L and are still within the scope of the present disclosure. 【0062】 The flexible facility 10 also includes a stainless-steel area 16 that includes at least one stainless-steel bioreactor 18. Each stainless-steel bioreactor 18 within the stainless-steel area 16 has a capacity greater than 5000 L. Additionally, as further described below, one or more of the at least one stainless-steel bioreactors 18 can be fluidly connected to one or more of the at least one single-use bioreactors 14. In another example, as shown in FIG. 1, the stainless-steel area 16 includes a plurality of stainless-steel bioreactors 20 each having a capacity of at least 10,000 L. 【0063】 In one example, at least one stainless steel bioreactor 18 is a large-scale stainless steel bioreactor and includes at least one valve assembly. In one example, and as described, at least one stainless steel bioreactor 18 may include a stationary steam sterilization bioreactor configured to hold a large volume of fluid, for example, a volume of up to 10,000 L or more. The at least one valve assembly of the stationary steam sterilization large-scale bioreactor enables the complete sterilization of the stationary steam sterilization large-scale bioreactor before starting cell culture (e.g., perfusion cell culture). Specifically, the at least one valve assembly may include a plurality of valves, and steam such as clean steam CS flows into one valve, then upward into other valves and reaches the ports. Further, after flowing into the first valve, the steam also flows downward into yet another valve, passes through a steam trap to sterilize the bioreactor, and creates a steam-sterilized aseptic environment. Further, as will be understood by those skilled in the art, process waste PW flows out of some of the valves during cleaning and is discharged to the drain. After use, the stationary steam sterilization bioreactor is stationary cleaned and the stationary steam sterilization process is performed again before further use. The at least one valve assembly also includes a port for coupling to a sterile connector valve assembly, such as an autoclaved sterile connector valve assembly, or other device. The sterile connector valve assembly enables the sterile connection of a single-use perfusion device or a reusable perfusion device to at least one stainless steel bioreactor 18, such as a stationary steam sterilization large-scale bioreactor. Further, the at least one valve assembly enables the sterile connection of a single-use bioreactor 14 to at least one stainless steel bioreactor 18. 【0064】 In another example, at least one stainless steel bioreactor 18 can include a side surface and a first valve assembly, such as an autoclaved valve assembly coupled to the side surface of the at least one stainless steel bioreactor 18. The autoclaved valve assembly can include a plurality of valves, and steam, such as clean steam CS, flows into a first valve, then upward into other valves, and reaches a port. Further, after flowing into the first valve, the steam flows downward through yet another valve, through a steam trap, sterilizes the at least one stainless steel bioreactor 18, and causes, for example, a stationary steam sterilization process before further use. In this example, there may be only at least one sterile connector coupled to a valve assembly, such as an autoclaved valve assembly. This autoclaved valve assembly enables connection to a factory-assembled and irradiated perfusion device or an autoclaved perfusion device, which can be, for example, a single-use perfusion device or a reusable perfusion device, via at least one sterile connector of the autoclaved valve assembly. Further, a plurality of perfusion devices can be aseptically coupled to the at least one stainless steel bioreactor 18 without additional steam sterilization. Further, the autoclaved valve assembly also enables connection of at least one single-use bioreactor 14 to the at least one stainless steel bioreactor 18. In yet another example, the at least one stainless steel bioreactor 18 can enable connection of an autoclaved perfusion device and in-place cleaning of the valve assembly without the use of a sterile connector. Instead, a hose assembly may be used as an alternative connector for coupling the at least one stainless steel bioreactor 18 to an autoclaved perfusion device and / or the at least one single-use bioreactor 14. 【0065】 As further shown in FIGS. 1 and 2A, in another example, the reconfigurable single-use area 12 and the stainless-steel area 16 are partitioned by a partition 22. In one example, the partition is a wall. In another example, the stainless-steel area 16 is completely enclosed and has separate air handling, as further described below. Further, a portion 24 of at least one stainless-steel bioreactor 18 penetrates the partition wall 22 and is fluidly connected via a tube 26 to one or more of at least one single-use bioreactor 14. As further described below, the fluid connection is configured to allow bypass of the stainless-steel area 16. 【0066】 In another example, at least one stainless-steel bioreactor 18 includes an adapter assembly such as a single-use adapter assembly and / or a Y-shaped assembly configured to be a sterile connector and / or coupled to a sterile connector. An adapter assembly such as a Y-shaped assembly includes a connector that is directly coupled to a sterile connector that is ultimately coupled to a pair of tubes such as tube 26. Each of one or more sterile connectors of the Y-shaped assembly is configured to be coupled to, for example, a reusable or single-use perfusion device in the reconfigurable single-use area 12 of FIGS. 1 and 2A, or at least one single-use bioreactor 14. This enables operationally coupling a plurality of perfusion devices or a plurality of single-use bioreactors 14 to, for example, at least one stainless-steel bioreactor 18. 【0067】 In another example, the portion 24 of at least one stainless-steel bioreactor 18 that penetrates the partition wall 22 includes one or more components including, but not limited to, a probe belt 27a, a perfusion connection 27b for perfusion, and a feed tank connection 27c as illustrated in FIG. 2B. 【0068】 As shown in FIGS. 1 and 2A, and in one example, the stainless steel area 16 includes one or more centrifuges 30. Further, the reconfigurable single-use area 12 also includes one or more centrifuges 32. In another example, the reconfigurable single-use area 12 includes one or more seed trains 34. In yet another example, the reconfigurable single-use area 12 includes one or more purification skids 36. In one example, and as shown in FIG. 2C, each of the one or more purification skids 36 includes one or more components independently selected from and / or including a chromatography skid 37a, a chromatography column 37b, a virus inactivation system 37c, a virus filtration system 37d, an ultrafiltration diafiltration system 37e, a bulk filtration system 37f, a conjugation tank system 37g, an oxidation tank system 37h, and a reduction tank system 37i. 【0069】 In some embodiments, the flexible facility 10 of the present disclosure exhibits or is configured with one or more of the following characteristics. 1) An extensive arrangement of reconfigurable space for cell culture processes (e.g., to accommodate high titers (e.g., titers of 1 to 15 g / L or more) at a 10,000 L scale, even higher titers (e.g., up to 30 g / L or more) at a 2,000 L scale). As part of a single therapeutic product manufacturing process, fed-batch culture and perfusion culture can be accommodated in one facility. 2) Include a stainless steel bioreactor 18 (e.g., a 10,000 L scale stainless steel production bioreactor) that maximizes purification capacity using commercially available chromatography resins with high binding capacity. 3) Include a stainless steel bioreactor 18 (e.g., a 10,000 L scale stainless steel production bioreactor) that is compatible with the typical maximum split ratio from a 2,000 L (N-1) bioreactor to a production bioreactor (N bioreactor). Exemplarily, with a 5:1 split ratio, a 2,000 L seed reactor in the (N-1) stage enables processing of a 10,000 L production bioreactor. 4) By means of a sterile connection between the stainless steel bioreactor 18 and the single-use bioreactor 14, the enhancement of the fed-batch operation is promoted. 5) Most of the stainless steel bioreactor 18 is arranged within the stainless steel area 16 with low-level air classification and low-level architectural finish, reducing construction costs and operating costs (such as air purification costs). 6) The downstream purification area is reconfigurable to accommodate a wide range of outputs from various upstream technologies, for example, by utilizing various easily movable production skids. The reconfigurable downstream purification area is configured to enable operation in batch mode or connection mode. In batch mode, it involves pool containers that separate various purification steps, and in connection mode, the pool containers are removed and direct connections between purification steps are used. 7) The reconfigurable downstream purification area is located adjacent to the reconfigurable seed train area within the reconfigurable single-use area 12. 【0070】 Each of the reconfigurable single-use area 12 and the stainless steel area 16 may include sub-areas that are partially or completely partitioned (for example, by partitions 22 such as walls or plastic sheets) from other parts of the reconfigurable single-use area 12 and the stainless steel area 16 in some embodiments. Further, the reconfigurable single-use area 12 may include one or more non-single-use devices (such as a stainless steel centrifuge) that are not bioreactors in some embodiments. Similarly, the stainless steel area 16 may include one or more non-stainless steel devices in some embodiments that are not bioreactors. 【0071】 The flexible facility 10 disclosed herein is useful for manufacturing at least one (for example, one, more than one (for example, two, three, four, etc.)) therapeutic product at a specific time. Exemplarily, the flexible facility 10 is adapted to manufacture various different therapeutic products simultaneously or sequentially by facilitating the compatibility of process-specific equipment while deploying common resources as much as possible. 【0072】 Non-limiting exemplary features Although not limiting, some exemplary embodiments / features of the present disclosure include the following. 1. A flexible facility for manufacturing at least one therapeutic product, comprising A reconfigurable single-use area including at least one single-use bioreactor, wherein one or more of the at least one single-use bioreactors have a capacity of at least 1000 L (e.g., 1000 L, 2000 L, 3000 L, 5000 L, at least 2000 L, at least 3000 L, at least 5000 L), and a reconfigurable single-use area; A stainless steel area including at least one stainless steel bioreactor, wherein each stainless steel bioreactor in the stainless steel area has a capacity greater than 5000 L, and a stainless steel area; Including One or more of the at least one stainless steel bioreactors are fluidly connected to one or more of the at least one single-use bioreactors, A flexible facility. 2. The flexible facility according to feature 1, wherein one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of the at least one single-use bioreactors have a capacity of 1000 L, 2000 L, 3000 L, or 5000 L. 3. The flexible facility according to feature 1 or 2, wherein one of the at least one single-use bioreactors has a capacity of 1000 L, 2000 L, 3000 L, or 5000 L. 4. The flexible facility according to any one of features 1 to 3, wherein none of the single-use bioreactors in the reconfigurable single-use area have a capacity greater than 2000 L. 5. One or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of the at least one single-use bioreactor has a capacity smaller than 2000L (e.g., 1000L, 500L, 50L), and is a flexible facility according to any one of features 1 to 4. 6. The reconfigurable single-use area includes one or more wave bioreactors (e.g., one or more wave bioreactors having a capacity of at least 50L (e.g., 50L)) and / or one or more seed bioreactors (e.g., one or more seed bioreactors having a capacity of at least 500L (e.g., 500L) or at least 2,000L (e.g., 2,000L)), and is a flexible facility according to any one of features 1 to 5. 7. The stainless-steel area includes a plurality of stainless-steel bioreactors, each having a capacity of at least 10,000L, and is a flexible facility according to any one of features 1 to 6. 8. Each stainless-steel bioreactor in the stainless-steel area has a capacity of 5000L to 20,000L, and is a flexible facility according to any one of features 1 to 7. 9. Each stainless-steel bioreactor in the stainless-steel area has a capacity of 5000L to 15,000L, and is a flexible facility according to any one of features 1 to 8. 10. Each stainless-steel bioreactor in the stainless-steel area has a capacity of 5000L to 10,000L, and is a flexible facility according to any one of features 1 to 9. 11. The stainless-steel area includes five 10,000L bioreactors, and is a flexible facility according to any one of features 1 to 10. 12. One or more of the at least one stainless-steel bioreactor are supported on the ground, and is a flexible facility according to any one of features 1 to 11. 13. One or more of the at least one stainless-steel bioreactor are suspended from the structure itself, and is a flexible facility according to any one of features 1 to 12. 14. The reconfigurable single-use area is a flexible facility as described in any one of features 1 to 13, including a reagent preparation area, a seed train area, and a purification area. 15. The reconfigurable single-use area is a flexible facility as described in any one of features 1 to 14, including a reagent preparation area, a seed train area, and a purification area, with one or more partitions that at least partially demarcate one or more of the reagent preparation area, the seed train area, and the purification area. 16. The seed train area includes one or more devices independently selected from a tank for storing a cell culture medium; a tank for storing an acidic solution, a basic solution, and a buffer; inoculation preparation equipment; a bioreactor (e.g., suitable for culturing cells), such as a locker-type bioreactor, a 500L bioreactor, and a 2000L single-use bioreactor; a tank suitable for containing cells; and a tank suitable for containing a product produced by the cells, in the flexible facility as described in feature 14 or 15. 17. The devices in the seed train area are in a single-use format, in the flexible facility as described in any one of features 14 to 16. 18. The stainless steel area is adjacent to the seed train area, in the flexible facility as described in any one of features 1 to 17. 19. The stainless steel area is adjacent to the purification area, in the flexible facility as described in any one of features 1 to 17. 20. The stainless steel area is adjacent to both the seed train area and the purification area, in the flexible facility as described in any one of features 1 to 17. 21. The stainless steel area further includes a recovery area, in the flexible facility as described in any one of features 1 to 20. 22. The stainless steel area is an unclassified manufacturing space, in the flexible facility as described in any one of features 1 to 21. 23. At least a part (e.g., the seed train area, the purification area) of the reconfigurable single-use area is a classified manufacturing space, in the flexible facility as described in any one of features 1 to 22. 24. The reconfigurable single - use area and the stainless - steel area are partitioned by a partition, and it is a flexible facility according to any one of Features 1 to 23. 25. The flexible facility according to Feature 24, wherein the partition is a plastic sheet. 26. The flexible facility according to Feature 25, wherein the partition is a wall. 27. The flexible facility according to Feature 26, wherein the stainless - steel area is completely enclosed and has separate air handling. 28. The flexible facility according to Feature 26 or 27, wherein a part of at least one stainless - steel bioreactor penetrates the wall and is fluid - connected to one or more of at least one single - use bioreactor via a pipe. 29. The flexible facility according to any one of Features 26 to 28, wherein at least 5% of the external container surface area of at least one stainless - steel bioreactor penetrates the wall and is fluid - connected to at least one of at least one single - use bioreactor via a pipe. 30. The flexible facility according to Feature 28 or 29, wherein the part penetrating the wall includes one or more components independently selected from a probe belt for enabling access to bioreactor instrumentation, a perfusion connection part, and a supply - liquid connection part. 31. The flexible facility according to any one of Features 1 to 30, wherein the fluid connection part includes a sterile connection module. 32. The flexible facility according to any one of Features 1 to 31, wherein the sterile connection module connects one or more seed - train bioreactors in the reconfigurable single - use area to one or more stainless - steel bioreactors in the stainless - steel area. 33. The flexible facility according to any one of Features 1 to 32, wherein the fluid connection part is configured to enable bypass of the stainless - steel area. 34. The flexible facility according to any one of Features 1 to 33, wherein the fluid connection part is configured to enable bypass of the stainless - steel area by using one or more valves. 35. The flexible facility according to any one of features 1 to 34, wherein the fluid connection part is detachable. 36. The flexible facility according to any one of features 1 to 35, wherein the fluid connection part can be moved between different bioreactors in the flexible facility. 37. The flexible facility according to any one of features 1 to 36, wherein the flexible facility includes a plurality of reconfigurable connection parts for connecting different devices (such as bioreactors, purification skids, etc.) in the flexible facility. 38. The flexible facility according to any one of features 1 to 37, wherein the reconfigurable single - use area and / or the stainless - steel area further includes one or more centrifuges. 39. The flexible facility according to any one of features 1 to 38, wherein the reconfigurable single - use area includes one or more seed trains. 40. The flexible facility according to any one of features 1 to 39, wherein the reconfigurable single - use area and / or the stainless - steel area includes one or more purification skids (for example, one or more purification skids including only single - use equipment; one or more purification skids including only stainless - steel equipment; one or more purification skids including both single - use equipment and stainless - steel equipment). 41. Each of the one or more purification skids includes one or more components independently selected from a chromatography skid, a chromatography column, a virus inactivation system, a virus filtration system, an ultrafiltration diafiltration system, and a bulk filtration system, for the flexible facility according to feature 40. 42. The flexible facility according to any one of features 1 to 41, further including one or more additional spaces independently selected from a utility space, a medium buffer facility, a dressing room, and a production unit. 43. The flexible facility according to any one of features 1 to 42, wherein the flexible facility is configured to produce one or more drug substances independently selected from monoclonal antibodies, BiTE (registered trademark) molecules, RNA - based drug substances, and fermentation products. 44. The flexible facility is configured to perform one or more processes independently selected from a mammalian cell culture fed-batch process, a mammalian cell culture perfusion process, a continuous mammalian cell culture process, a microbial fermentation fed-batch process, and any combination of the foregoing, the flexible facility according to any one of features 1 to 43. 45. The flexible facility can be constructed in a series of stages, the flexible facility according to any one of features 1 to 44. 46. The flexible facility is extensible to add additional product lines, the flexible facility according to any one of features 1 to 45. 47. The stainless-steel bioreactor within the stainless-steel area contains only one therapeutic product-producing cell line, the flexible facility according to any one of features 1 to 46. 48. The stainless-steel bioreactor within the stainless-steel area contains more than one therapeutic product-producing cell line, the flexible facility according to any one of features 1 to 46. 49. The single-use bioreactor within the reconfigurable single-use area contains only one therapeutic product-producing cell line, the flexible facility according to any one of features 1 to 48. 50. The single-use bioreactor within the reconfigurable single-use area contains more than one therapeutic product-producing cell line, the flexible facility according to any one of features 1 to 48. 51. Each therapeutic product-producing cell line is independently selected from prokaryotic cell lines and eukaryotic cell lines, the flexible facility according to any one of features 47 to 50. 52. Each therapeutic product-producing cell line is independently selected from prokaryotic cell lines, the flexible facility according to any one of features 47 to 50. 53. Each therapeutic product-producing cell line is independently selected from mammalian cell lines, the flexible facility according to any one of features 47 to 50. 54. Each therapeutic product production cell line is independently selected from cell lines capable of producing viral therapeutics (e.g., oncolytic anti-cancer viruses, viral vectors for gene therapy and / or viral immunotherapy), and is a flexible facility according to any one of features 47 to 50. 55. Each therapeutic product production cell line is independently selected from CHO cell lines, and is a flexible facility according to any one of features 47 to 50. 【0073】 Definition: As used herein, the terms "a" and "an" mean "one or more" unless otherwise indicated. Further, "one or more" and "at least one" are used interchangeably herein. Further, unless the context requires a different interpretation, singular terms include the plural, and plural terms include the singular. 【0074】 As used herein, the term "bioreactor" means any vessel useful for the growth of a cell culture (e.g., a mammalian cell culture or a bacterial cell culture). "Bioreactor" as used herein encompasses the term "fermenter" (i.e., a vessel useful for the growth of a bacterial cell culture, typically including a more powerful stirrer and an increased gas flow compared to a vessel used for the growth of a mammalian cell culture). Non-limiting examples of bioreactors include stirred tank type, air-lift type, fiber type, microfiber type, hollow fiber type, ceramic matrix type, fluidized bed type, fixed bed type, and / or jet loop type bioreactors. In some embodiments, an exemplary bioreactor can perform one or more (e.g., one, two, three, all) of the following steps: supply of nutrients and / or carbon source, injection of appropriate gas (e.g., oxygen, etc.), inflow and outflow of fermentation or cell medium, separation of gas phase and liquid phase, maintenance of temperature, maintenance of oxygen and CO2 levels, maintenance of pH level, agitation (e.g., stirring), and / or washing / sterilization. Unless otherwise indicated by the context, a bioreactor can be suitable for a batch process, a semi-fed batch process, a fed batch process, a perfusion process, and / or a continuous fermentation process. Any suitable bioreactor diameter can be used. Unless otherwise indicated by the context, in some embodiments, a bioreactor can have a volume of 100 mL to 50,000 L. Unless otherwise indicated, a bioreactor can be of any size as long as it is useful for culturing cells, and typically, a bioreactor is sized to be suitable for the volume of the cell culture to be grown therein. In non-limiting embodiments and unless otherwise indicated by the context, a bioreactor can be at least 1 liter (L), or 2, 5, 10, 50, 100, 200, 250, 500, 1,000, 1,500, 2,000, 2,500, 5,000, 8,000, 10,000, 12,000 liters or more, or any volume in between. The internal conditions of a bioreactor, including but not limited to pH and temperature, can be controlled during the culture period.A person skilled in the art can recognize and select a bioreactor suitable for use in the flexible facility disclosed herein based on relevant considerations. 【0075】 As used herein, the terms "cell culture" or "culturing" refer to the growth and proliferation of cells outside of a multicellular organism or tissue. Suitable culture conditions for mammalian cells and bacterial cells are known in the art. (See, for example, Animal cell culture: A Practical Approach, D. Rickwood, ed., Oxford University Press, New York (1992).) Mammalian cells may be cultured in suspension or while attached to a solid culture medium. In some embodiments, a fluidized bed bioreactor, a hollow fiber bioreactor, a roller bottle, a shake flask, and / or a stirred tank bioreactor may be used for cell culture, with or without microcarriers. In some embodiments, a 500 L to 2000 L bioreactor is used for cell culture (e.g., as part of a seed train). In some embodiments, a 1000 L to 2000 L bioreactor is used for cell culture (e.g., as part of a seed train). 【0076】 As used herein, the term "cell culture medium" (also referred to as "medium", "culture medium", "cell culture medium", "tissue culture medium", etc.) refers to any nutrient solution used to grow cells, such as bacterial cells or mammalian cells. Cell culture media generally provide one or more of the following components: an energy source (e.g., in the form of a carbohydrate such as glucose); one or more essential amino acids (e.g., all 20 basic amino acids in addition to cysteine); vitamins and / or other organic compounds typically required at low concentrations; lipids or free fatty acids; and trace elements such as inorganic compounds or naturally occurring elements typically required at very low concentrations, e.g., in the micromolar range. As used herein, cell culture media include any nutrient solution that is typically used in and / or is known to be used in any cell culture process, including but not limited to batch culture, extended batch culture, fed-batch culture, and / or perfusion culture, or continuous culture of cells. 【0077】 As used herein, "fed-batch culture" refers to a form of suspension culture, specifically a method of culturing cells in which additional components are provided to the culture medium at one or more time points after the start of the culture process. The components supplied typically include nutrient supplements for the cells that are depleted during the culture process. Additionally or alternatively, the additional components may include supplemental components (e.g., cell cycle inhibitory compounds, etc.). In some embodiments, the fed-batch cell culture medium composition may be more concentrated or higher in concentration than the basal cell culture medium composition. The fed-batch culture may be stopped at some point, and the cells and / or components in the medium may be harvested and optionally purified. 【0078】 As used herein, a "perfusion" cell culture medium refers to a cell culture medium that is typically used in a cell culture maintained by a perfusion or continuous culture method and is a complete cell culture medium sufficient to support the cell culture during this process. In some embodiments, the perfusion cell culture medium composition can be more concentrated or at a higher concentration than the basal cell culture medium composition to accommodate the method used to remove spent medium. In some embodiments, the perfusion cell culture medium can be used in both the growth and production phases. 【0079】 As used herein, a "production" cell culture medium refers to a cell culture medium that is typically used in a cell culture during the transition when exponential growth ends and protein production becomes dominant (i.e., the "transition" and / or "production" phases), and is a complete cell culture medium sufficient to maintain the desired cell density, viability, and / or productivity titer during this period. 【0080】 As used herein, a "therapeutic product" is a biological product for use in a subject (e.g., a human) in relation to the prevention, diagnosis, cure, or alleviation of a disease, disorder, defect, or injury. Non-limiting examples of therapeutic products include monoclonal antibodies, BiTE® molecules, RNA-based prodrugs, cell therapies, and fermentation products. 【0081】 Seed train area A seed train area, such as the area of the single-use area 12 within the flexible facility 10 where one or more seed trains 34 are disposed, can house equipment suitable for cell culture. Equipment for cell culture includes tanks for storing cell culture media; tanks for acidic, basic, and buffer solutions; inoculation preparers; rocker-type bioreactors, 500L bioreactors, and 2000L single-use bioreactors, such as the single-use bioreactor 14 (e.g., suitable for culturing cells) bioreactors; tanks suitable for containing cells or products produced by the cells; centrifuges, such as centrifuge 32; pumps; and other equipment useful for product recovery, but are not limited thereto. In some embodiments, the equipment within the seed train area is in single-use form. 【0082】 In some embodiments, the seed train area demonstrates the flexibility of the manufacturing facility of the present disclosure. For example, in some configurations where a stainless steel bioreactor 18 is also used, the seed train can include up to the N-1 bioreactor and the bioreactor unit including the N-1 bioreactor. In some embodiments, the product of the N-1 bioreactor is transferred to the stainless steel N bioreactor. In this configuration, the equipment for product recovery is bypassed and the equipment within the stainless steel area is prioritized. 【0083】 In some configurations where the stainless steel section is bypassed, the largest bioreactor unit is considered the N bioreactor (e.g., a 500L or 2000L N bioreactor, etc.), and the equipment within the seed train area for product recovery is utilized. 【0084】 In some embodiments, the seed train area includes one or more flasks, culture bags, and bioreactor units suitable for culturing cells. In some embodiments, the cell culture units within the seed train area increase in size within the seed train to accommodate larger cell cultures. In some embodiments, the seed train can include one wave bioreactor and two seed bioreactors. In some embodiments, the seed train can include one 50L wave bioreactor, one 500L single-use bioreactor, and one 2000L single-use bioreactor, and a 2000L single-use bioreactor such as the single-use bioreactor 14 is optionally configured for perfusion culture (such as ATF perfusion culture). In some embodiments, there may also be a parallel set of increasingly larger bioreactors, and each parallel set can include cells expressing the same product or cells expressing different products. 【0085】 In some embodiments, the seed train area can include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bioreactors, or multiples thereof when manufacturing multiple products. In some embodiments, each bioreactor within the seed train area is suitable for one or more process methods independently selected from a batch process, a semi-fed batch process, a fed batch process, a perfusion process, and / or a continuous process. In some embodiments, the bioreactor is a stirred tank reactor. In some embodiments, the bioreactor is an air-lift reactor. In some embodiments, the bioreactor can have a capacity of from about 100 milliliters to about 5,000 liters. Non-limiting exemplary bioreactor capacities include 100 milliliters, 250 milliliters, 500 milliliters, 750 milliliters, 1 liter, 2 liters, 3 liters, 4 liters, 5 liters, 6 liters, 7 liters, 8 liters, 9 liters, 10 liters, 15 liters, 20 liters, 25 liters, 30 liters, 40 liters, 50 liters, 60 liters, 70 liters, 80 liters, 90 liters, 100 liters, 150 liters, 200 liters, 250 liters, 300 liters, 350 liters, 400 liters, 450 liters, 500 liters, 550 liters, 600 liters, 650 liters, 700 liters, 750 liters, 800 liters, 850 liters, 900 liters, 950 liters, 1000 liters, 1500 liters, 2000 liters, 2500 liters, 3000 liters, 3500 liters, 4000 liters, 4500 liters, and 5000 liters. Generally, the bioreactor units within the seed train area are smaller than the bioreactor units within the stainless steel area. In a particular configuration, each seed train bioreactor is at least one-half, at least one-third, at least one-fourth, or at least one-fifth of the stainless steel production bioreactor. In some embodiments, the seed train section can accommodate four 2,000-liter containers for single-use technology operations. 【0086】 In some embodiments, the seed train area complies with appropriate manufacturing processes and biological safety standards. In some embodiments, the seed train area complies with the standards of Biosafety Level 1 (BSL1), Biosafety Level 2 (BSL2), Biosafety Level 3 (BSL3), or Biosafety Level 4 (BSL4). 【0087】 In some embodiments, the seed train area can include sub-compartments, and each sub-compartment can be used to perform different functions or aspects to support the cell culture production process. By way of example, in some embodiments, the seed train area can include a sub-compartment that houses one or more bioreactors and a sub-compartment that houses equipment for product recovery. 【0088】 Single-use production area In some embodiments where the stainless-steel area 16 is bypassed, the reconfigurable single-use area 12 includes one or more production bioreactors. The flexible facility 10 is easily expandable and scalable and can produce completely different therapeutic products within the same flexible facility 10 using different production bioreactors within the reconfigurable single-use area 12. 【0089】 For example, in some embodiments, in a modular unit configured as a first bioreactor module, the user can manufacture one type of therapeutic product, such as a monoclonal antibody product derived from a mammalian cell line. In a second modular unit, the user can manufacture a completely different therapeutic product, such as a microbial product. In some embodiments, the flexible facility 10 of the present disclosure can support multiple product lines simultaneously using the production bioreactors in one or both of the single-use production area 12 and the stainless-steel area 16. Further, in some embodiments, a flexible facility, such as the flexible facility 10 of the present disclosure, is expandable to add additional product lines. 【0090】 Stainless steel area In some embodiments, stainless steel section 16 houses equipment suitable for cell culture. Equipment suitable for cell culture includes, but is not limited to, bioreactors (e.g., suitable for culturing cells), tanks (e.g., suitable for containing cells, media, or products produced by cells), centrifuges, pumps, and other equipment useful for product recovery, as well as cleaning-in-place (CIP) systems. The CIP unit is typically a modular skid and has several tanks for holding cleaning fluids (e.g., caustic solutions and / or bleach), pumps, and sensors for sending the cleaning fluids to the appropriate tanks to be cleaned. 【0091】 In some embodiments, the stainless steel section 16 includes one or more stainless steel bioreactor units, such as a stainless steel bioreactor 18 suitable for culturing cells. The bioreactor unit can perform one or more of the following, namely, supply of nutrients and / or carbon source, injection of appropriate gas (e.g., oxygen), flow of cell culture medium, separation of gas phase and liquid phase, maintenance of growth temperature, maintenance of pH level, agitation (e.g., stirring), and / or washing / sterilization (e.g., one, two, three, four, all, etc.). In some embodiments, the stainless steel section can include 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, or 100 or more bioreactors. In some embodiments, each bioreactor within the stainless steel area 16 is suitable for one or more process methods independently selected from a batch process, a semi-fed batch process, a fed batch process, a perfusion process, and / or a continuous process. In some embodiments, at least one stainless steel bioreactor 18 is a stirred tank reactor. In some embodiments, at least one stainless steel bioreactor 18 is an airlift reactor. In some embodiments, at least one stainless steel bioreactor 18 can have a capacity of 1000 liters to 50,000 liters. Non-limiting examples include capacities of 1000 liters, 1500 liters, 2000 liters, 2500 liters, 3000 liters, 3500 liters, 4000 liters, 4500 liters, 5000 liters, 6000 liters, 7000 liters, 8000 liters, 9000 liters, 10,000 liters, 15,000 liters, 20,000 liters, or 50,000 liters. 【0092】 In some embodiments, approximately 95% of the stainless-steel bioreactor is in an unclassified space within the flexible facility 10, and a small portion (about 5%) of the stainless-steel bioreactor 18 is exposed in the classified and reconfigurable single-use area 12 of the flexible facility 10. In some embodiments, the exposed portion of the stainless-steel bioreactor 18 includes a probe belt 27a of the tank to enable access to tank instrumentation, a perfusion connection 27b, and any other connection 27c for adding feed liquid and solution to the tank, including connections for pH control, nutrient feed (medium), or other additions (i.e., addition of antifoaming agent). 【0093】 In some embodiments, the exposed portion 24 of the stainless-steel bioreactor 18 is directly exposed into the classified space by, for example, one or more connections made in one or more lines within the classified space that may enter the tank by means of the probe belt 27a or through piping at other tank locations (e.g., by addition of medium, etc.). 【0094】 In some embodiments, the stainless-steel area 16 is designed to accommodate several bioreactors 18. In some embodiments, the stainless-steel area 16 can accommodate five 10,000-liter containers for mammalian cell lines. 【0095】 In some embodiments, the stainless-steel area 16 includes five or more 10,000-liter containers and recovery equipment. In some embodiments, the stainless-steel area 16 includes five 10,000-liter containers and recovery equipment. In some embodiments, the bioreactors within the stainless-steel area are configured to produce monoclonal antibody products derived from mammalian cell lines. 【0096】 In some embodiments, one or more of the stainless-steel bioreactors 18 within the stainless-steel area 16 can be ground-mounted reactors. In some embodiments, one or more of the stainless-steel bioreactors 18 can be suspended from the structure itself. 【0097】 In some embodiments, the stainless-steel area 16 complies with appropriate manufacturing processes and biological safety standards. In some embodiments, the stainless-steel area 16 complies with the standards of Biosafety Level 1 (BSL1), Biosafety Level 2 (BSL2), Biosafety Level 3 (BSL3), or Biosafety Level 4 (BSL4). 【0098】 In some embodiments, the stainless-steel area 16 can include sub-compartments, and each sub-compartment can be used to perform different functions or aspects to assist the cell culture production process. By way of example, in some embodiments, the stainless-steel area 16 can include a sub-compartment for accommodating one or more bioreactors, a sub-compartment for accommodating equipment for product recovery, a sub-compartment for inoculation materials, and a sub-compartment for cleaning and decontaminating equipment and operators handling such equipment. 【0099】 In some embodiments, different production bioreactors within the stainless-steel area 16 can be used to produce completely different therapeutic products in the same flexible facility, such as the flexible facility 10. For example, in some embodiments, in a modular unit configured as a first bioreactor module, a user can manufacture one type of therapeutic product, such as a monoclonal antibody product derived from a mammalian cell line. In a second modular unit, the user can manufacture a completely different therapeutic product, such as a microbial product. 【0100】 Downstream processing area Standard downstream processing (DSP) units generally are an assembly of chromatography, mixing equipment, and filtration equipment. In non-limiting embodiments of the present disclosure, filtration equipment within the downstream processing area can include ultrafiltration equipment, microfiltration equipment, virus filtration equipment, and sterilization equipment, such as, for example, a pre-virus separation subunit and a post-virus separation subunit. Virus reduction can occur throughout an overall purification process of a typical mammalian cell-derived protein. For example, in some embodiments, virus reduction can occur within an entire mammalian process through a virus reduction chromatography step, virus inactivation using a mixing vessel, and / or virus filtration. The post-virus separation subunit houses equipment and utilities suitable for any one of the following, namely, ultrafiltration (tangential flow filtration), normal filtration, chromatography, formulation, titration, mixing, concentration, buffer exchange, and container filling and freezing of bulk drug substance. 【0101】 Non-limiting exemplary purification areas can include a stirred tank with an inlet conduit for a crude extract product from a separation module; an outlet conduit with a positive displacement pump connected to an inlet conduit of a first chromatography column operated by a computerized unit to which the conduit is also connected; an outlet conduit from a chromatography column connected to an inlet conduit of a stirred tank, the outlet conduit of which with a positive displacement pump is connected to an inlet conduit of a second chromatography column operated by a computerized unit to which the conduit is also connected; and an outlet conduit from a chromatography column connected to an inlet conduit of a stirred tank, the outlet conduit of which is, in turn, discontinuously connected to an inlet of a lyophilizer with an outlet. 【0102】 Classification level Different areas within the flexible facility 10 (e.g., the reconfigurable single-use area 12 and the stainless steel area 16, and parts thereof, etc.) can have different classification levels based on grading criteria. For example, in some embodiments, the different areas can have different classification levels based on grading criteria set by the regulatory agency of the jurisdiction in which the flexible facility 10 is regulated. In some embodiments, the different areas can have different classification levels based on grading criteria set by the regulatory agency of the jurisdiction where the flexible facility 10 is located. In some embodiments, the different areas can have different classification levels based on grading criteria set by the regulatory agency of the jurisdiction where the therapeutic products produced in the flexible facility 10 are sold. 【0103】 In some embodiments, the different areas can have different classification levels based on grading criteria set by the US Food and Drug Administration, or based on grading criteria set by the Rules Governing Medicinal Products in the European Union Volume 4 EU Guidelines to Good Manufacturing Practice Medicinal Products for Human and Veterinary Use, supplemented by EudraLex, Annex 1 Manufacture of Sterile Medicinal Products in the European Union. 【0104】 Control system In some embodiments, samples from cell cultures can be monitored and evaluated using any of the analytical techniques well-known in the art. In some embodiments, various parameters can be monitored over the course of the culture, including but not limited to the quality and characteristics of recombinant proteins and media. Samples can be taken and monitored intermittently at a desired frequency, including continuous monitoring, real-time monitoring, or near real-time monitoring. 【0105】 In some embodiments, the seed train equipment, the stainless steel bioreactor, and the purification equipment may each include sensors for monitoring reaction parameters and / or product quality parameters. Parameters to be monitored may include, but are not limited to, conductivity, temperature, pH, oxygen, and CO2. The sensors may be any type of invasive sensor well known in the art for monitoring these parameters, and the sensors contact the process fluid. Further, in some embodiments, the sensors may be non-invasive optical chemical sensors such as those described in, for example, U.S. Patent Nos. 6,673,532 and 7,041,493 and U.S. Patent Application Publication No. 20110065084. Further, spectrometers well known in the art can be used in the seed train equipment, the production bioreactor, and / or the purification equipment to monitor the product stream and / or the input to each piece of equipment. Parameters measured by such spectrometers may include, but are not limited to, absorbance, multi-angle light scattering, fluorescence, Raman scattering, circular dichroism, and infrared spectroscopic characteristics. 【0106】 In some embodiments, the equipment and processes within each area are computer controlled and monitored via a computer system and corresponding software, thereby enabling automation of the system. The equipment within each area may include data collection sensors and control units that can communicate with a central controller via either a wired connection or a wireless connection. 【0107】 In some embodiments, each area may further include an electronic control panel that can also be used to monitor, collect, and control the equipment and the processes occurring therein. The central controller may also include data storage for storing data, as well as applications and / or operating software for the central controller and system operation. The software may include a graphical user interface (GUI) that can be displayed on a workstation. The GUI may display several different screens for setting, monitoring, and controlling the equipment and the overall manufacturing process. 【0108】 Thus, in some embodiments, such software can be used to control procedures within the area and optionally can be customized via a central computer and / or an electronic control panel. The procedures are strategies for performing specific processes within the area (i.e., materials produced or to be produced by one execution of a batch process). In some embodiments, the process can be a series of chemical, physical, or biological activities for the conversion, transport, or storage of materials or energy. 【0109】 In some embodiments, the aseptic connection module connects one or more seed train bioreactors within a reconfigurable single-use area to one or more stainless steel bioreactors within a stainless steel area. 【0110】 In some embodiments, the aseptic connection module connects a recovery unit to one or more downstream purification units. 【0111】 Cell line In some embodiments, each bioreactor within the flexible facility 10 of the present disclosure is suitable for culturing prokaryotic or eukaryotic cells. In some embodiments, at least one bioreactor within the flexible facility 10 is suitable for culturing prokaryotic cells. In some embodiments, at least one bioreactor within the flexible facility 10 is suitable for culturing eukaryotic (e.g., mammalian) cells. In some embodiments, at least one bioreactor within the flexible facility 10 is suitable for culturing cells capable of producing viral therapeutics (e.g., oncolytic anti-cancer viruses, viral vectors for gene therapy and / or viral immunotherapy). In some embodiments, at least one bioreactor within the flexible facility 10 is suitable for culturing prokaryotic cells capable of producing viral therapeutics (e.g., oncolytic anti-cancer viruses, viral vectors for gene therapy and / or viral immunotherapy). In some embodiments, at least one bioreactor within the flexible facility 10 is suitable for culturing eukaryotic (e.g., mammalian) cells capable of producing viral therapeutics (e.g., oncolytic anti-cancer viruses, viral vectors for gene therapy and / or viral immunotherapy). 【0112】 Examples of cells suitable for culturing in the bioreactor of the flexible facility 10 of the present disclosure include, but are not limited to, bacterial cells (e.g., Escherichia coli (E. coli), Pichia pastoris), yeast cells (e.g., Saccharomyces cerevisiae, Trichoderma reesei), plant cells, insect cells (e.g., Sf9), Chinese hamster ovary cells (CHO, and any genetically modified or derived CHO cell line), mouse cells (e.g., mouse embryonic fibroblasts, cells derived from a mouse cancer model), human cells (e.g., cells from any tissue or organ, cells from a cancer or other disease cell line, stem cells, hybridoma cells, or other genetically modified or hybrid cells). Host cells suitable for use in a bioreactor housed within the flexible facility of the present disclosure include, for example, those commercially available from a culture collection such as DSMZ (Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH, Braunschweig, Germany) or American Type Culture Collection (ATCC). 【0113】 In some embodiments, the cell is a bacterial cell or a prokaryotic cell. In some embodiments, the prokaryotic cell is selected from Gram-positive cells (e.g., cells of the genus Bacillus (e.g., Bacillus subtilis (e.g., B. subtilis 3NA and B. subtilis 168), Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus natto, or Bacillus megaterium), the genus Streptomyces, the genus Streptococcus, the genus Staphylococcus, or the genus Lactobacillus). In some embodiments, the prokaryotic cell is a Gram-negative cell, e.g., Salmonella spp. or Escherichia coli, e.g., TG1, TG2, W3110, DH1, DHB4, DH5a, HMS174, HMS174(DE3), NM533, C600, HB101, JM109, MC4100, XL1-Blue, and Origami cells, and cells derived from E. coli B strains such as, for example, BL-21 or BL21(DE3). The prokaryotic cell or bacterial cell can be obtained from a commercial vendor. For example, Bacillus cells can be obtained, for example, from Bacillus Genetic Stock Center, Biological Sciences 556, 484 West 12th Avenue, Columbus Ohio 43210-1214. 【0114】 In some embodiments, the cells are eukaryotic cells such as mammalian cells. The mammalian cells can be, for example, cell lines or cell lineages of human, rodent, or bovine origin. Examples of such cells, cell lines, or cell lineages include, but are not limited to, mouse myeloma (NSO) cell line, Chinese hamster ovary (CHO) cell line, FIT1080, H9, HepG2, MCF7, MDBK Jurkat, NIH3T3, PC12, BF1K (baby hamster kidney cells), VERO, SP2 / 0, YB2 / 0, Y0, C127, L cells, COS (e.g., COS1 and COS7), QC1-3, HEK-293, VERO, PER.C6, HeLa, EB1, EB2, EB3, oncolytic, or hybridoma cell lines. In some embodiments, the mammalian cells are CHO cell lines. In some embodiments, the mammalian cells are CHO cells. In some embodiments, the mammalian cells are selected from CHO-K1 cells, CHO-K1 SV cells, DG44 CHO cells, DUXB11 CHO cells, CHOS cells, CHO GS knockout cells, CHO FUT8 GS knockout cells, CHOZN cells, and CHO-derived cells. The CHO GS knockout cells (e.g., GSKO cells, etc.) are, for example, CHO-K1 SV GS knockout cells. Further, the CHO FUT8 knockout cells are, for example, Potelligent® CHOK1 SV (Lonza, Inc.). In some embodiments, the eukaryotic cells can be, for example, avian cells, cell lines, or cell lineages such as EBx® cells, EB14, EB24, EB26, EB66, or EBv13. 【0115】 In some embodiments, any mammalian cell line can be used in the bioreactor included within the flexible facility 10 of the present disclosure. As non-limiting examples, a wide variety of mammalian cell lines suitable for growth in culture are available from the American Type Culture Collection (Manassas, Va.) and vendors. Non-limiting examples of cell lines commonly used in the art include monkey kidney CVl cell line transformed by SV40 (COS-7, ATCC CRL 1651); human fetal kidney cell line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., 1977, J. Gen Virol. 36:59); baby hamster kidney cells (BHK, ATCC CCL 10); mouse Sertoli cells (TM4, Mather, 1980, Biol. Reprod. 23:243-251); monkey kidney cells (CVl ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical cancer cells (HELA, ATCC CCL 2); dog kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human hepatocellular carcinoma cells (Hep G2, HB 8065); mouse mammary carcinoma (MMT 060562, ATCC CCL51); TRI cells (Mather et al., 1982, Annals N.Y Acad. Sci. 383:44-68); MRC5 cells or FS4 cells; mammalian myeloma cells, and many other cell lines, as well as Chinese hamster ovary (CHO) cells. 【0116】 CHO cells containing CHOK1 cells (ATCC CCL61) are widely used for producing complex recombinant proteins. In some embodiments, dihydrofolate reductase (DHFR)-deficient mutant cell lines (Urlaub et al., 1980, Proc Natl Acad Sci USA 77:4216-4220), DXB11 and DG-44 are desirable CHO host cell lines because an efficient DHFR selectable and amplifiable gene expression system enables high-level expression of recombinant proteins in these cell lines (Kaufman R.J., 1990, Meth Enzymol 185:537-566). Also included are glutamine synthetase (GS) knockout CHOK1SV cell lines that utilize methionine sulfoximine (MSX) selection based on glutamine synthetase (GS). Other suitable CHO host cells for use in the flexible facility bioreactors of the present disclosure include, but are not limited to, the following (the numbers in parentheses are ECACC accession numbers). CHO(85050302), CHO(protein-free)(00102307), CHO-K1(85051005), CHO-K1 / SF(93061607), CHO / dhFr-(94060607), CHO / dhFr-AC-free(05011002), and RR-CHOKI(92052129). 【0117】 Large-scale production of commercially useful proteins can be carried out in suspension culture. Accordingly, mammalian host cells used to generate the recombinant mammalian cells described herein can be adapted for growth in suspension culture, but this is not required. A variety of host cells adapted for growth in suspension culture are known and include mouse myeloma NS0 cells, and CLIO cells derived from the CFIO-S, DG44, and DXB11 cell lines. Other suitable cell lines include mouse myeloma SP2 / 0 cells, baby hamster kidney BF1K-21 cells, human PER.C6® cells, human fetal kidney F1EK-293 cells, and cell lines derived from or modified from any of the cell lines disclosed herein. 【0118】 In some embodiments, the eukaryotic cell is selected from lower eukaryotic cells such as, for example, yeast cells (e.g., Pichia (e.g., Pichia pastoris, Pichia methanolica, Pichia kluyveri, and Pichia angusta), Komagataella (e.g., Komagataella pastoris, Komagataella pseudopastoris, or Komagataella phaffii), Saccharomyces cells (e.g., Saccharomyces cerevisae, Saccharomyces kluyveri, Saccharomyces uvarum), Kluyveromyces cells (e.g., Kluyveromyces lactis, Kluyveromyces marxianus), Candida cells (e.g., Candida utilis, Candida cacaoi, Candida boidinii), Geotrichum cells (e.g., Geotrichum fermentans), Hansenula polymorpha, Yarrowia lipolytica, or Schizosaccharomyces pombe). In some embodiments, the eukaryotic cell is selected from Pichia pastoris strains. Non-limiting examples of Pichia pastoris strains include X33, GS115, KM71, KM71H, and CBS7435. 【0119】 In some embodiments, the eukaryotic cell is selected from fungal cells (e.g., cells of the genus Aspergillus (e.g., A. niger, A. fumigatus, A. orzyae, A. nidula, etc.), the genus Acremonium (e.g., A. thermophilum, etc.), the genus Ketomiopsis (e.g., C. thermophilum, etc.), the genus Chrysosporium (e.g., C. thermophile, etc.), the genus Cordyceps (e.g., C. militaris, etc.), the genus Collimonas, the genus Kutzneromyces, the genus Fusarium (e.g., F. oxysporum, etc.), the genus Glomerella (e.g., G. graminicola, etc.), the genus Hypocrea (e.g., Η. jecorina, etc.), the genus Magnaporthe (e.g., Μ. orzyae, etc.), the genus Meliophthora (e.g., Μ. thermophile, etc.), the genus Nectria (e.g., Ν. heamatococca, etc.), the genus Neurospora (e.g., Ν. crassa, etc.), the genus Penicillium, the genus Sporothrix (e.g., S. thermophile, etc.), the genus Tuberomyces (e.g., Τ. terrestris, Τ. heterothallica, etc.), the genus Trichoderma (e.g., Τ. reesei, etc.), or the genus Verticillium (e.g., V. dahlia, etc.)). 【0120】 In some embodiments, the eukaryotic cell is selected from insect cells (e.g., Sf9, Mimic™ Sf9, Sf21, High Five™ (BT1-TN-5B1-4), or BT1-Ea88 cells, etc.), algal cells (e.g., Amphora, Bacillariophyceae, Dunaliella, Chlorella, Chlamydomonas, Cyanophyta (cyanobacteria), Nannochloropsis, Spirulina, or Ochromonas, etc.), and plant cells (e.g., cells from monocotyledonous plants (e.g., corn, rice, wheat, or Setaria, etc.) or dicotyledonous plants (e.g., cassava, potato, soybean, tomato, tobacco, alfalfa, Physcomitrella patens or Arabidopsis, etc.)). 【0121】 In some embodiments, the cells express or produce a product, such as a recombinant therapeutic product. Examples of therapeutic products produced by the cells, as described in more detail below, include antibody molecules (e.g., monoclonal antibodies, bispecific antibodies), antibody mimetics (polypeptide molecules that specifically bind to an antigen but are not structurally related to antibodies, such as DARPins, affibodies, adnectins, or IgNARs, etc.), fusion proteins (e.g., Fc fusion proteins, chimeric cytokines), other recombinant proteins (e.g., glycosylated proteins, enzymes, hormones), viral therapeutics (e.g., anti-cancer oncolytic viruses, viral vectors for gene therapy and viral immunotherapy), cell therapeutics (e.g., pluripotent stem cells, mesenchymal stem cells, and adult stem cells), vaccines or lipid-encapsulated particles (e.g., exosomes, virus-like particles), RNA (e.g., siRNA) or DNA (e.g., plasmid DNA), antibiotics, or amino acids, but are not limited thereto. In some embodiments, the flexible facility according to the present disclosure can be used for the production of biosimilars. In some embodiments, the flexible facility according to the present disclosure can be used for the production of therapeutic products having various complexities, which may include post-translational modifications such as glycosylation. 【0122】 In some embodiments, the flexible facility according to the present disclosure enables the production of eukaryotic cells (e.g., mammalian cells or lower eukaryotic cells (e.g., yeast cells or filamentous fungal cells, etc.)), or prokaryotic cells (e.g., Gram-positive bacteria or Gram-negative bacteria, etc.), and / or products of eukaryotic or prokaryotic cells (e.g., proteins, peptides, antibiotics, amino acids, nucleic acids (e.g., DNA or RNA, etc.) synthesized on a large scale by eukaryotic cells). 【0123】 Culture methods Various methods that can be used in commercial processes for the production of recombinant proteins by mammalian cell culture include, but are not limited to, batch culture, fed-batch culture, and perfusion culture. 【0124】 Batch culture is a discontinuous method in which cells are grown in a fixed volume of culture medium for a short period and then the entire volume is harvested. Cultures grown using the batch method go through an increase in cell density until they reach the maximum cell density, after which the viable cell density decreases as the medium components are consumed and the levels of metabolic by-products (such as lactate and ammonia) accumulate. Harvesting is typically done when the maximum cell density (e.g., 5×10 6 cells / mL or more, depending on the medium composition, cell line, etc.) is reached. The batch process is the simplest culture method, but the viable cell density is limited by the availability of nutrients, and when the cells reach their maximum density, the culture declines and production decreases. In batch culture, the production period cannot be extended because the accumulation of waste and depletion of nutrients lead to a rapid decline in the culture, which is usually about 3 to 7 days. 【0125】 Fed-batch culture improves the batch process by providing a bolus or continuous supply of medium to replenish the consumed medium components. Fed-batch culture receives additional nutrients throughout the operation, resulting in a higher cell density (e.g., >10 - 30×10 6have the potential to achieve increased viable cell densities (cells / mL) and increased productivity titers. Unlike batch processes, by manipulating the feeding strategy and media composition, a two-phase culture can be created and maintained to distinguish between a cell growth phase (the growth phase) for achieving a desired cell density and a period during which cell growth has ceased or slowed (the production phase). Therefore, fed-batch cultures have the potential to achieve higher productivity titers compared to batch cultures. Typically, the batch method is used during the growth phase and the fed-batch method is used during the production phase, although the fed-batch feeding strategy can be used throughout the process. However, unlike batch processes, the bioreactor volume is a limiting factor that restricts the amount of feed. Also, as with batch processes, the accumulation of metabolic by-products leads to culture decline, which typically limits the production phase to a period of about 10 - 21 days. Fed-batch cultures are non-continuous, and harvest is typically performed when the metabolic by-product level or culture viability reaches a predetermined level. Fed-batch cultures can produce more recombinant protein compared to batch cultures without feeding. (See, for example, U.S. Patent No. 5,672,502.) 【0126】 Perfusion methods offer the possibility of improvement over batch and fed-batch methods by adding fresh media during culture while simultaneously removing spent media. Perfusion culture is a culture in which the cell culture receives fresh perfusion feed media while simultaneously removing spent media. A typical perfusion culture begins with a batch culture startup that lasts for 1 or 2 days, after which continuous, stepwise, and / or intermittent addition of fresh feed media to the culture and simultaneous removal of spent media are performed, retaining cells and additional high molecular weight compounds such as proteins (based on the molecular weight cut-off of the filter) throughout the growth and production phases of the culture. Various methods such as sedimentation, centrifugation, or filtration can be used to remove spent media while maintaining cell density. A non-limiting example of a filtration method is tangential flow filtration. Tangential flow is maintained by pumping the media through a hollow fiber filter module. See, for example, U.S. Patent No. 6,544,424; Furey, 2002, Gen. Eng. News. 22(7):62 - 63. 【0127】 Perfusion can be continuous, stepwise, intermittent, or any combination or all of these. The perfusion rate can range from less than the working volume per day to many times the working volume. The cells are retained in the culture, and the spent medium removed is substantially cell-free or has significantly fewer cells compared to the culture. Recombinant proteins expressed by cell culture can also be retained in the culture. 【0128】 In typical large-scale commercial cell culture strategies, the aim is to achieve high cell densities of 60 - 90(+)×10 6 cells / mL, where approximately one-third to more than one-half of the reactor volume is biomass. In perfusion cultures, very high cell densities of >1×10 8 cells / mL have been achieved, and even higher densities are predicted. A potential advantage of the perfusion process is the ability to maintain production cultures over longer periods compared to batch or fed-batch culture methods. However, to maintain long-term perfusion cultures, especially in the case of cultures at high cell densities that require more nutrients, it is necessary to increase the preparation, use, storage, and disposal of the medium. All of these result in increased production costs compared to batch and fed-batch methods. Furthermore, high cell densities can cause problems such as maintaining dissolved oxygen levels during production, as well as problems associated with increased aeration, including more oxygen supply and more carbon dioxide removal, which may require more foaming and defoaming strategy changes, and the work required to remove excess cell material can lead to product losses, potentially negating the advantage of increased titer due to increased cell mass, and can cause problems in the recovery and downstream processes. 【0129】 An alternative large-scale cell culture strategy suitable for use in the flexible facility 10 of the present disclosure is a combination of fed-batch feeding during the growth phase and continuous perfusion during the production phase. In some embodiments, a fed-batch culture with bolus feeding is used to maintain cell culture during the growth phase. Thereafter, perfusion feeding can be used during the production phase. In some embodiments, perfusion is initiated when the cells reach the production phase. 【0130】 Therapeutic product In the flexible facility according to the present disclosure, various therapeutic products can be produced. The polypeptides and proteins of interest are part of the therapeutic products and can be for scientific or commercial purposes, including protein-based therapeutics. The proteins of interest include, but are not limited to, secreted proteins, non-secreted proteins, intracellular proteins, or membrane-bound proteins. The polypeptides and proteins of interest can be produced by recombinant animal cell lines using cell culture methods and can be referred to as "recombinant proteins". The expressed proteins can be produced intracellularly or secreted into the culture medium from which they can be recovered and / or harvested. The term "isolated protein" or "isolated recombinant protein" refers to the polypeptide or protein of interest that is separated and purified from proteins or polypeptides or other contaminants that would interfere with its therapeutic, diagnostic, prophylactic, research, or other use. The proteins of interest include, but are not limited to, proteins that exert a therapeutic effect by binding to targets such as those listed below (including targets derived from them, targets related to them, and variants thereof). 【0131】 The protein of interest includes, but is not limited to, "antigen-binding protein". "Antigen-binding protein" refers to a protein or polypeptide that contains an antigen-binding region or antigen-binding moiety having affinity for another molecule (antigen) to which it binds. Examples of antigen-binding proteins include, but are not limited to, antibodies, peptibodies, antibody fragments, antibody derivatives, antibody mimetics, fusion proteins (including, for example, single-chain variable fragments (scFv), double-chain (bivalent) scFv, and IgGscFv (see, for example, Orcutt et al., 2010, Protein Eng Des Sel 23:221-228), heteroIgG (see, for example, Liu et al., 2015, J Biol Chem 290:7535-7562), mutains, and XmAb® (Xencor, Inc., Monrovia, CA). Also included are bispecific T cell engagers (BiTE®), such as bispecific T cell engagers having an extension such as extended half-life, e.g., HLE BiTE, HeteroIg BITE, and others, chimeric antigen receptors (CAR, CAR T), and T cell receptors (TCR). 【0132】 In some embodiments, the target protein may include colony-stimulating factors such as granulocyte colony-stimulating factor (G-CSF). Such G-CSF agents include, but are not limited to, Neupogen® (filgrastim) and Neulasta® (pegfilgrastim). Also, for example, Epogen® (epoetin alpha), Aranesp® (darbepoetin alpha), Dynepo® (epoetin delta), Mircera® (methoxypolyethylene glycol-epoetin beta), Hematide®, MRK-2578, INS-22, Retacrit® (epoetin zeta), Neorecormon® (epoetin beta), Silapo® (epoetin zeta), Binocrit® (epoetin alpha), epoetin alpha Hexal, Abseamed® (epoetin alpha), Ratioepo® (epoetin theta), Eporatio® (epoetin theta), Biopoin® (epoetin theta), epoetin alpha, epoetin beta, epoetin zeta, epoetin theta, epoetin delta, epoetin omega, epoetin iota, tissue plasminogen activator, GLP-1 receptor agonist, and variants or analogs thereof, and erythropoiesis-stimulating agent (ESA) such as any biosimilar of the foregoing are also included. 【0133】 In some embodiments, the target protein binds to one or more of the following, alone or in any combination: CD proteins including, but not limited to, CD3, CD4, CD5, CD7, CD8, CD19, CD20, CD22, CD25, CD30, CD33, CD34, CD38, CD40, CD70, CD123, CD133, CD138, CD171, and CD174; HER receptor family proteins including, for example, HER2, HER3, HER4, and EGF receptor; EGFRvIII; cell adhesion molecules including, for example, LFA-1, Mol, p150,95, VLA-4, ICAM-1, VCAM, and alpha v / beta 3 integrin; growth factors including, but not limited to, vascular endothelial growth factor ( "VEGF"); VEGFR2; growth hormone; thyroid stimulating hormone; follicle stimulating hormone; luteinizing hormone; growth hormone releasing factor; parathyroid hormone; Mullerian duct inhibitory substance; human macrophage inflammatory protein (MIP-1-alpha); erythropoietin (EPO); nerve growth factors such as NGF-beta; platelet-derived growth factor (PDGF); fibroblast growth factors including, for example, aFGF and bFGF; epithelial growth factor (EGF); Cripto; transforming growth factor (TGF) including, in particular, TGF-alpha and TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta4, or TGF-beta5; insulin-like growth factors-I and -II (IGF-I and IGF-II); des(1-3)-IGF-I (brain IGF-I); and osteogenic factor; insulin and insulin-related proteins including, but not limited to, insulin, insulin A chain, insulin B chain, proinsulin, and insulin-like growth factor binding proteins; coagulation and coagulation-related proteins including, for example, factor VIII, tissue factor, von Willebrand factor, protein C, alpha-1-antitrypsin, plasminogen activators such as urokinase and tissue plasminogen activator ( "t-PA"), bombesin, thrombin, thrombopoietin, and thrombopoietin receptor; colony stimulating factors (CSF) including, in particular, M-CSF, GM-CSF, and G-CSF; other blood and serum proteins including, but not limited to, albumin, IgE, and blood group antigens;Receptors and receptor-related proteins including, for example, the flk2 / flt3 receptor, the obesity (OB) receptor, the growth hormone receptor, and the T cell receptor; Neurotrophic factors including, but not limited to, bone-derived neurotrophic factor (BDNF) and neurotrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6); Relaxin A chain, relaxin B chain, and prorelaxin; Interferons including, for example, interferon-alpha, -beta, and -gamma; Interleukins (IL) such as, for example, IL-1 to IL-10, IL-12, IL-15, IL-17, IL-23, IL-12 / IL-23, IL-2Ra, IL1-R1, IL-6 receptor, IL-4 receptor and / or IL-13 receptor, IL-13RA2, or IL-17 receptor, IL-1RAP, etc.; Virus antigens including, but not limited to, AIDS envelope virus antigen; Lipoproteins; Calcitonin; Glucagon; Atrial natriuretic factor; Lung surfactant; Tumor necrosis factor-alpha and -beta; Enkephalinase; BCMA; IgKappa; ROR-1; ERBB2; Mesothelin; RANTES (regulated upon activation, normal T cell expressed and secreted); Mouse gonadotropin-related peptide; DNase; FR-alpha; Inhibin; And activin; Integrin; Protein A or D; Rheumatoid factor; Immunotoxin; Bone morphogenetic protein (BMP); Superoxide dismutase; Surface membrane protein; Decay-accelerating factor (DAF); AIDS envelope; Transport protein; Homing receptor; MIC (MIC-a, MIC-B); ULBP 1-6; EPCAM; Addressin; Regulatory protein; Immune adhesion molecule; Antigen-binding protein; Somatropin; CTGF; CTLA4; Eotaxin-1; MUC1; CEA; c-MET; Claudin-18; GPC-3; EPHA2; FPA; LMP1; MG7; NY-ESO-1; PSCA; Ganglioside GD2; Ganglioside GM2; BAFF; OPGL (RANKL); Myostatin; Dickkopf-1 (DKK-1); Ang2; NGF; IGF-1 receptor; Hepatocyte growth factor (HGF); TRAIL-R2; c-Kit; B7RP-1; PSMA; NKG2D-1; Programmed cell death protein 1 and ligand; PD1 and PDL1;Mannose receptor / hCGβ; Hepatitis C virus; Mesothelin dsFv[PE38] conjugate; Legionella pneumophila (lly); IFN gamma; Interferon gamma-induced protein 10 (IP10); IFNAR; TALL-1; Thymic stromal lymphopoietin (TSLP); Proprotein convertase subtilisin / Kexin type 9 (PCSK9); Stem cell factor; Flt-3; Calcitonin gene-related peptide (CGRP); OX40L; α4β7; Platelet specific (platelet glycoprotein IIb / IIIb (PAC-1); Transforming growth factor beta (TFGβ); Zona pellucida sperm-binding protein 3 (ZP-3); TWEAK; Platelet-derived growth factor receptor alpha (PDGFRα); Sclerostin; and any biologically active fragment or variant of any of the foregoing. 【0134】 In some embodiments, the protein of interest is abciximab, adalimumab, adecatumumab, aflibercept, alemtuzumab, alirocumab, anakinra, atacicept, basiliximab, belimumab, bevacizumab, biosozumab, blinatumomab, brentuximab vedotin, brodalumab, canertinib mertansine, canakinumab, cetuximab, certolizumab pegol, conatumumab, daclizumab, denosumab, eculizumab, edrecolomab, efalizumab, epratuzumab, etanercept, evolocumab, galiximab, ganitumab, gemtuzumab, golimumab, ibritumomab tiuxetan, infliximab, ipilimumab, leridani mab, lumiliximab, ixekizumab, mapatumumab, motesanib diphosphate, muromonab-CD3, natalizumab, nesiritide, nimotuzumab, nivolumab, ocrelizumab, ofatumumab, omalizumab, oprelvekin, palivizumab, panitumumab, pembrolizumab, pertuzumab, paxalisizumab, ranibizumab, rilotumumab, rituximab, romiplostim, romosozumab, sargramostim, tocilizumab, tositumomab, trastuzumab, ustekinumab, vedolizumab, visilizumab, volociximab, zanubrutinib, and zalutumumab, and any biosimilar of any of the foregoing. 【0135】 The proteins of interest may also include genetically engineered receptors such as chimeric antigen receptors (CARs or CAR-Ts) and T cell receptors (TCRs), as well as other proteins including antigen-binding molecules that interact with their target antigens. By incorporating an antigen-binding molecule that interacts with its target antigen, a CAR can be engineered to bind to an antigen (e.g., a cell surface antigen, etc.). A CAR typically incorporates an antigen-binding domain (such as an scFv) along with one or more co-stimulatory ("signaling") domains and one or more activation domains. 【0136】 All documents or portions of documents cited herein, including but not limited to patents, patent applications, papers, books, and academic articles, are hereby expressly incorporated by reference into this specification. The embodiments described in the present disclosure can be combined with one or more other embodiments of the present disclosure, unless the context clearly indicates otherwise. 【0137】 The disclosed subject matter is not intended to be limited in scope by the specific embodiments described herein. Instead, it is intended as an illustrative, non-limiting example of the individual aspects of the present disclosure. Functionally equivalent methods and components are within the scope of the present disclosure. Indeed, various modifications of the disclosed subject matter will be apparent to those skilled in the art from the foregoing and the appended drawings. Such modifications are intended to be within the scope of the disclosed subject matter. 【0138】 The descriptions of the various embodiments and / or examples of the disclosed subject matter are presented for illustrative purposes and are not intended to be exhaustive or limiting in any way. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terms used herein are chosen to best explain the principles of the embodiments, the practical application, or a technical improvement found in the marketplace, and / or to enable those skilled in the art to understand the disclosed subject matter.

Claims

[Claim 1] A flexible facility for manufacturing at least one therapeutic product, A reconfigurable single-use area comprising at least one single-use bioreactor, wherein one or more of the at least one single-use bioreactors have a capacity of at least 1000 L, and the reconfigurable single-use area comprises A stainless steel area comprising at least one stainless steel bioreactor, wherein each stainless steel bioreactor within the stainless steel area has a capacity greater than 5000 L, Includes, One or more of the at least one stainless steel bioreactors are fluidly connected to one or more of the at least one single-use bioreactors. Flexible facilities. [Claim 2] The flexible facility according to claim 1, wherein one or more of the at least one single-use bioreactors have a capacity of at least 2,000 L. [Claim 3] The flexible facility according to claim 1 or 2, wherein the stainless steel area includes a plurality of stainless steel bioreactors, each having a capacity of at least 10,000 L. [Claim 4] The flexible facility according to claim 1, wherein the reconfigurable single-use area and the stainless steel area are separated by a partition. [Claim 5] The flexible facility according to claim 4, wherein the partition is a wall. [Claim 6] The flexible facility according to claim 5, wherein the stainless steel area is completely enclosed and has separate air handling. [Claim 7] The flexible facility according to claim 5, wherein a portion of the at least one stainless steel bioreactor penetrates the wall and is fluidly connected via a pipe to one or more of the at least one single-use bioreactors. [Claim 8] The flexible facility according to claim 7, wherein the portion penetrating the wall includes one or more components independently selected from a probe belt, a perfusion connector, and a supply fluid tank connector. [Claim 9] The flexible facility according to any one of claims 1, 2, 4 to 8, wherein the fluid connection is configured to allow bypassing of the stainless steel area. [Claim 10] The flexible facility according to any one of claims 1, 2, 4 to 8, wherein the reconfigurable single-use area and / or the stainless steel area further comprises one or more centrifuges. [Claim 11] The flexible facility according to any one of claims 1, 2, 4 to 8, wherein the reconfigurable single-use area includes one or more seed trains. [Claim 12] The flexible facility according to any one of claims 1, 2, 4 to 8, wherein the reconfigurable single-use area further comprises one or more purified skids. [Claim 13] The flexible facility according to claim 12, wherein each of the one or more purification skids comprises one or more components independently selected from a chromatography skid, a chromatography column, a virus inactivation system, a virus filtration system, an ultrafiltration diafiltration system, and a bulk filtration system. [Claim 14] The flexible facility according to any one of claims 1, 2, 4 to 8, further comprising one or more additional spaces independently selected from utility spaces, culture medium buffer facilities, changing rooms, and production units. [Claim 15] The flexible facility according to any one of claims 1, 2, 4 to 8, wherein the flexible facility is configured to produce one or more therapeutic products independently selected from monoclonal antibodies, BiTE® molecules, RNA-based active pharmaceutical ingredients, cell therapies, and fermentation products. [Claim 16] The flexible facility according to any one of claims 1, 2, 4 to 8, wherein the flexible facility is configured to carry out one or more processes independently selected from a mammalian cell culture fed batch process, a mammalian cell culture perfusion process, a continuous mammalian cell culture process, a microbial fermentation fed batch process, and any combination thereof.

Images