Midscale model for organic growth and phasing

Abstract

To provide a system for the industrial growth of biologics.SOLUTION: The present disclosure is directed to the organic growth and phasing design for a system for the industrial growth of biologics. A system may include a number of subsystems, such as a buffer distribution subsystem, a media preparation subsystem, a bioreactor subsystem, a harvest subsystem, and / or a purification subsystem. The subsystems may be highly interconnected for flexible process flow design. Additionally, the subsystems may be constructed with a total output capacity with a total number of equipment stations and operated at a lower output capacity with fewer than the total equipment stations to maintain headroom for growth in phases.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] Related Applications

[0001] This application is based on and claims priority to U.S. Provisional Patent Application No. 62 / 679,076, filed June 1, 2018, which is incorporated herein by reference. [Background technology]

[0002]

[0002] The pharmaceutical industry has traditionally been heavily focused on so-called blockbuster drugs that can require significant investment in permanent facilities dedicated to a single process. Once established, the efficiency of a single process flow can provide a strong return on initial investment.

[0003]

[0003] However, market factors demand industrial processes with increased flexibility at reduced cost. In particular, the growing demand for specialized drugs has highlighted the shortcomings of facilities designed for a single major product line. Pressures for quality control and extensive development and improvement can require multiple modifications or iterations of a particular process flow.

[0004]

[0004] For example, various biological cell culture processes can be used to extract useful organic components, such as proteins, enzymes, metabolites, or other biomass. Such processes are often time consuming and require significant capital investment to build a single facility to carry out the culture and growth process. In many cases, an existing facility previously configured for one product may not be easily or quickly reconfigured to accommodate a new product iteration in a test or production capacity.

[0005]

[0005] Furthermore, when the production stage is reached, existing facilities are not easily reconfigured to scale up from initial small product yields. However, traditional high-yield facilities can become inefficient when exposed to rapidly changing product cycles and may not be suitable for the early stages of development.

[0006]

[0006] In view of the above, there is a need for a flexible production line that can be quickly adapted and expanded based on needs, allowing for fast reconfiguration, greater throughput, and greater product diversity. Furthermore, there is also a need for flexibility in both process and equipment design, as well as the facilities and buildings designed to contain the process. In short, there is a need for organism growth and phasing for the industrial growth of biopharmaceuticals. Summary of the Invention

[0007]

[0007] Generally, the present disclosure is directed to a system for the growth of a biopharmaceutical. The system includes a bioreactor subsystem with at least two bioreactors and at least two harvest subsystems. Each harvest subsystem, either alone or in combination, is in fluid communication with each of the bioreactors in the bioreactor subsystem.

[0008] In some embodiments, the system includes a flow control assembly positioned between the at least two bioreactors and the at least two harvest subsystems. The flow control assembly may be configured to selectively control a flow rate of fluid from the first bioreactor to the first harvest subsystem and from the first bioreactor to the second harvest subsystem. The flow control assembly may be further configured to control a flow rate of fluid from the second bioreactor to the first harvest subsystem and from the second bioreactor to the second harvest subsystem.

[0009] In some embodiments, the system further includes at least one purification subsystem. Each purification subsystem, alone or in combination, is in fluid communication with each of the harvest subsystems. In some embodiments, at least one of the purification subsystems is configured to produce a different biologic than at least one other purification subsystem.

[0010] In some embodiments, a system as disclosed herein can be configured to process more than 100 batches per year.

[0011] In some embodiments, at least one of the bioreactors is 6000 L or larger. In some embodiments, the bioreactor subsystem comprises a seed train of fed-batch bioreactors and / or at least one perfusion seed train. In some embodiments, the perfusion seed train may feed at least one fed-batch bioreactor.

[0011]

[0012] The disclosure is further directed to at least one buffer dilution device that receives at least two inputs (i) and (ii), where (i) is a buffer concentrate source stream and (ii) is a water for injection source stream. The system further includes at least one buffer dilution device that delivers a buffer mixture including (i) and (ii) to a buffer mixture distribution manifold. The at least one buffer mixture distribution manifold may be used to control parameters of the buffer mixture delivered independently to each of the multiple destinations, the parameters including at least one of flow rate or concentration. The buffer mixture is not stored prior to delivery to any of the multiple destinations.

[0012]

[0013] In one embodiment, the system for buffer distribution further comprises a first buffer dilution device. The buffer mixture flowing out of the first buffer dilution device is delivered as inflow (i) to at least one downstream buffer dilution device. The inflow (i) of any buffer dilution device downstream of the first buffer dilution device may be partially diverted to another buffer dilution device or to at least one buffer mixture distribution manifold. The at least two buffer mixture distribution manifolds distribute buffers of different concentrations.

[0013]

[0014] In some embodiments, the at least two buffer dilution devices deliver approximately the same concentration of buffer to the at least one buffer distribution manifold.

[0015] In some embodiments, the at least one buffer destination includes a media preparation subsystem, a cell culture subsystem, a bioreactor subsystem, a harvest subsystem, and / or a purification subsystem.

[0014]

[0016] The present disclosure is further directed to a system for growing biologics generally including a perfusion seed train, a bioreactor subsystem including at least one bioreactor fed in whole or in part from the perfusion seed train, and at least two harvest subsystems, each of which may be configured, alone or in combination, to receive the effluent of each of the bioreactors in the bioreactor subsystem.

[0015]

[0017] The disclosure is further generally directed to a building including any of the systems described above. For example, in one embodiment, the building may include at least two purification subsystems, each including three chromatography column stations and three chromatography skid stations in a first room, a chromatography column station and a chromatography skid station in a second adjacent room, and a maintenance area accessible from either the first room or the second room. The maintenance area may be accessible from the first and second rooms by airlocks. It is further accessible from the rest of the building by another airlock.

[0016]

[0018] In some embodiments, the second chamber further comprises a filling device.

[0019] In some embodiments, the bioreactor subsystem comprises multiple bioreactor stations in one room, some or all of the stations being filled with bioreactors. In some embodiments, at least one bioreactor station is not filled with a bioreactor. In some embodiments, at least one of the bioreactor stations is not filled with a bioreactor, and at least one of the bioreactor stations that is filled with a bioreactor is configured to maintain fluid communication with a flow control assembly. The flow control assembly can be configured to selectively control the flow rate of fluid from any one of the at least two bioreactor stations to any one of the at least two harvest subsystems.

[0017]

[0020] In some embodiments, a building provided as herein may include logistics corridors, personnel areas, packaging areas, shipping areas, and / or receiving areas and may be configured to meet Leadership in Energy and Environmental Design (LEED) certification standards. The building may further include a plurality of motion, temperature, and / or humidity sensors distributed on, in, and / or around equipment within the building, and a monitoring system configured to track the output of the plurality of sensors and generate records and / or alarms in response to the output deviating from a predetermined range.

[0018]

[0021] The disclosure is further generally directed to a process for growing a biologic, the process including growing a cell mass, harvesting a material containing (i) the cells or (ii) a component of the cells, passing a first portion of the harvested material to a first purification subsystem, passing a second portion of the harvested material to a second purification subsystem, producing a first product via the first purification subsystem, and producing a second product via the second purification subsystem.

[0019]

[0022] A full and enabling disclosure of the present disclosure is set forth in more detail in the remainder of the specification, including reference to the accompanying drawings. [Brief description of the drawings]

[0020] [Figure 1]

[0023] FIG. 1 illustrates an exemplary layout of a facility provided as herein; [Diagram 2]

[0024] FIG. 1 shows a schematic of a buffer distribution subsystem. [Diagram 3]

[0025] FIG. 1 shows a schematic of the media preparation subsystem. [Figure 4]

[0026] FIG. 1 shows a schematic of a bioreactor subsystem. [Diagram 5]

[0027] FIG. 1 shows a schematic of a collection subsystem. [Figure 6]

[0028] FIG. 1 shows a schematic of the purification subsystem. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021]

[0029] Repeat use of reference characters in the present specification and drawings is intended to represent same or analogous features or elements of the invention.

[0030] It will be appreciated by those skilled in the art that this discussion is merely a description of exemplary embodiments and is not intended to limit the broad aspects of the present disclosure.

[0022]

[0031] In general, the present disclosure is directed to a system for the growth of biologics. As used herein, growing a biologic generally refers to culturing, providing, and processing biological material to achieve a particular purpose. In some cases, the purpose is to grow the grown material. In some cases, the desired product is the acquisition of the material itself. For example, a particular cell culture may be desired for its therapeutic use, and the growing process expands a given amount of the cell culture to harvest, wash, and possibly package the desired cells. In other cases, some by-product of the biomass is desired. For example, in some cases, a protein product of a particular cell culture is desired, and the growing process expands a given amount of the cell culture to extract, purify, and possibly package the protein product.

[0023]

[0032] The system may be described as a collection of organized subsystems. These subsystems may be organized within a single building or facility. If necessary, the facility may be built to meet standards for controlled and / or classified spaces, such as those defined by ISO 14644-1, FED STD 209E, and / or EU Good Manufacturing Practice (GMP) standards. Merely by way of example, disposable equipment may be mentioned. It is understood that reusable equipment falls within the scope of the present disclosure.

[0024]

[0033] A flexible process according to the present disclosure may be housed in a facility 000 as shown in FIG. 1. The exemplary facility may be divided into three sub-facilities: an upstream sub-facility 001, a downstream sub-facility 002, and an operator sub-facility 003. There may be more than one of any of the sub-facilities. The sub-facilities may be built adjacently or stacked, such as on different floors of a building. For example, the upstream sub-facility 001 may be on the second floor, while the downstream sub-facility 002 may be on the first floor. Alternatively, the sub-facilities may be housed in different adjacent buildings. In some examples, the upstream and downstream sub-facilities may be located in different subdivisions (e.g., rooms or floors) of a building, while the operator sub-facility 003 may be located in, around, between, under, and / or beside the upstream and / or downstream sub-facility.

[0025]

[0034] In one embodiment, as shown in FIG. 1, the upstream sub-facility 001 includes a buffer distribution subsystem 100, a media preparation subsystem 200, and a bioreactor subsystem 300, the downstream sub-facility 002 includes two harvest subsystems 400 and two purification subsystems 500, and the operational sub-facility 003 may include, for example, a logistics corridor, personnel areas, packaging areas, shipping areas, and / or receiving areas.

[0026]

[0035] In some embodiments, facility 000 is located within the same building as other facilities used for the production of other products. For example, multiple facilities 000 may be co-located for the production of multiple biologics.

[0027]

[0036] Of particular advantage, a single facility 000 may further produce any number of biologics within its contained subsystems. For example, the subsystems within a facility may be highly interconnected. In one embodiment shown in FIG. 1, the effluent of the bioreactor subsystem 300 may be in independent fluid communication with both of the harvest subsystems 400. Additionally, any one of the harvest streams from any one of the harvest subsystems 400 may be in fluid communication with any one or both of the purification subsystems 500. The buffer mixture generated in the buffer distribution subsystem 100 may be directed to any one of the other subsystems. In this manner, a single facility 000 may provide maximum flexibility for configuring one or more desired process flows.

[0028]

[0037] As used herein, fluid communication refers to a direct and intentional connection through several different conduits, allowing fluid to flow from one point to another. The fluid communication may be regulated by any number of valves, manifolds, or controllers.

[0029]

[0038] For example, in one embodiment, the flow control assembly may provide a framework for interconnected fluid communication. In some cases, the flow control assembly may be a system of pipes. Some embodiments may further employ any number of valves, diverters, throttles, meters, or manifolds, each controllable in a manual and / or automated fashion. In this manner, the flow control assembly may facilitate fluid communication between any number of subsystems or locations having subsystems of the facility 000.

[0030]

[0039] The flow control assembly can be selective in its flow control. For example, when a flow control assembly is used to provide fluid communication between a single bioreactor subsystem 300 having two or more bioreactors and two or more harvest subsystems 400, the flow control assembly individually selectively controls the effluent from any subset of the bioreactors to direct each subset to one or more destination harvest subsystems individually selected for effluent, also with individually customized flow parameters. In this manner, the flow control assembly can provide a very flexible fluid communication network between all or any subset of the bioreactors in the bioreactor subsystem 300 to all or any subset of the multiple harvest subsystems 400. In some embodiments, a similar flow control assembly can enable selective and individually controlled distribution of buffers to any number of subsystems, anywhere in the facility 000.

[0031]

[0040] Flexible embodiments of facility 000, in some examples, include a fluid connection network between multiple subsystems. The subsystems can be located and relocated to different locations within the facility, and the fluid connection network infrastructure (including, for example, flow control assemblies) can enable rapid reconnection of equipment.

[0032]

[0041] Advantageously, a fluid connection network is created to service more subsystems than it was originally constructed for. For example, a facility 000 as in FIG. 1 may include a fluid connection network that is fully configured to provide fluid connections between one bioreactor subsystem 300 and two harvest subsystems 400, but is initially operated with one harvest subsystem 400.

[0033]

[0042] Such interconnected facilities 000 may operate in parallel, in whole or in part, e.g., an entire product train may or may not be fully parallelized. In one embodiment, several parallel bioreactor subsystems 300 may feed a single harvest subsystem 400 that feeds several parallel purification subsystems 500. In another example, a bioreactor subsystem 300 feeds multiple harvest subsystems 400 that feed multiple purification subsystems 500. In this manner, a single facility 000 may produce several different products or several grades of the same product, e.g., a single harvest subsystem 400 may provide the same harvest culture to two purification subsystems 500, one optimized for quality and one optimized for yield. Similarly, various test scenarios may be run in parallel to observe different process effects on the very same source batch.

[0034]

[0043] Facilities prepared according to the present disclosure have the ability to be optimized to maximize throughput, and subsystems can be independently configured to achieve any desired throughput requirements. For example, different purification trains can be tuned at different operating throughputs to minimize excess product. Additionally, some embodiments may purposely operate below full capacity to temporarily lower operating costs when maximum throughput is not required.

[0035]

[0044] In some embodiments, the total output of facility 000 may be greater than about 50 batches per year, such as greater than about 75 batches per year, such as greater than about 100 batches per year, such as greater than about 150 batches per year, such as greater than about 300 batches per year. Generally, however, facility 000 may be scaled to produce less than about 1000 batches per year.

[0036]

[0045] The facility 000 may require an operating personnel, perhaps in some cases a large number of operating personnel. However, in some examples, the facility 000 may be partially or fully automated. For example, predictive analytics technology (PAT) may enable streamlined maintenance and monitoring procedures. Several sensors, such as vibration, noise, humidity, temperature, or power consumption sensors, may provide a depiction of the operational health of the facility 000 to the monitoring system. For example, vibration and temperature sensors may be mounted on the equipment casing. If any output of any of the sensors deviates from a predefined range, or alternatively, a range interpreted by the monitoring system as normal or stable, the monitoring system may generate an alarm to indicate that a particular area of ​​the facility 000, such as in a particular subsystem, requires attention. In this manner, the total operating personnel required may be reduced.

[0037]

[0046] Advantageously, the flexibility of the facility 000 provided herein allows for quick recovery from unplanned equipment downtime. For example, if one of the two harvesting subsystems 400 goes offline, the interconnected fluid communication network allows the effluent of the bioreactor subsystem 300 to be quickly and easily redirected to the second harvesting subsystem 400.

[0038]

[0047] Due to the highly flexible nature of the facility 000 provided in accordance with the present disclosure, the facility may be built to be energy efficient. For example, total utility consumption may be optimized to be efficient by selectively operating various subsystems only when necessary to meet demand. In some embodiments, the building containing the facility may be built to meet various standards, such as Leadership in Energy and Environmental Design (LEED) certification standards.

[0039]

[0048] In one embodiment, the system includes a buffer distribution subsystem 100 shown in Figure 2. The buffer solution may be a salt solution used to alter or maintain a given pH level in a target environment, such as, for example, a bicarbonate solution. For example, a buffer may be added to a bioreactor feed medium to obtain and / or maintain a particular pH. The area in which the buffer is prepared may be a controlled airspace, such as ISO 8 / Grade C / Class 100,000.

[0040]

[0049] The buffer may be mixed and subsequently stored in a large tank prior to delivery to the target environment. However, in some embodiments, the buffer concentrate may be mixed, for example, in a small disposable mixer (SUM) 102 or a large SUM 104. If used, the small and / or large SUMs may be of various sizes. For example, a small SUM may be greater than about 100 L, such as greater than about 500 L, such as greater than about 1000 L. However, in general, a small SUM may be smaller than about 3000 L, such as less than about 1000 L. A large SUM for buffer concentrate preparation may be greater than about 500 L, such as greater than about 1000 L, such as greater than about 2000 L, such as greater than about 3000 L. However, in general, a large SUM may be smaller than about 4000 L, such as less than about 3000 L.

[0041]

[0050] The contents of the buffer may be selected appropriately for a given application. In some embodiments, the treatment solution is a buffer such as Tris, Tricine, HEPES, MOPS, PIPES, TAPS, Bicine, BES, TES, cacodylic acid, MES, acetate, MKP, ADA, ACES, glycinamide, and acetamidoglycine, or acetic acid.

[0042]

[0051] In some embodiments, a bicarbonate buffer may be desirable. For example, sodium carbonate and sodium bicarbonate may be provided in dry form and mixed with water for injection. The dry contents may be deposited by automated equipment or manually.

[0043]

[0052] The buffer may be stored in a concentrated form in the holding tank 106. The holding tank may be of any useful configuration. For example, the holding tank may be a stainless steel vessel. In other embodiments, the holding tank may include one or more disposable bags.

[0044]

[0053] The stored concentrate may then be pumped to a buffer dilution device 112. The buffer dilution device 112 may receive a buffer concentrate stream 108 and sterile water for injection stream 110. In some embodiments, the buffer mixture stream 114 proceeds directly to a buffer mixture distribution manifold 116 after a single dilution stage.

[0045]

[0054] In some embodiments, the buffer mixture stream is directed into a second buffer dilution device 112 that dilutes the buffer mixture stream into a second buffer mixture stream 118. The second buffer mixture stream 118 can be distributed by a second buffer mixture distribution manifold 120. In this manner, the buffer distribution subsystem 100 can produce and distribute buffer solutions having any number of different concentrations.

[0046]

[0055] In some embodiments, two or more buffer dilution devices 112 produce the same concentration of buffer solution. Such embodiments are more resistant to downtime due to system failure or maintenance and may also allow for increased flexibility to fluctuating buffer demands.

[0047]

[0056] In one embodiment, the buffer mixture distribution manifold 116 controls the flow rate of the buffer mixture to any number of destinations. In some embodiments, the buffer mixture distribution manifold is automated to individually control the flow rate of the buffer mixture to multiple destinations, and optionally respond to sensor feedback indicative of a target parameter, such as the pH of the destination environment.

[0048]

[0057] The buffer mixture distribution manifold may optionally be centrally located to allow compact and efficient routing of buffer distribution lines to any number of destinations. In some embodiments, some destinations may also be manifolds intended for local distribution. For example, a central manifold may pump buffer to a particular room in a processing facility, and smaller manifolds within this room may distribute the buffer to devices within the room that require the buffer solution.

[0049]

[0058] In some embodiments, a system provided in accordance with the present disclosure includes a media preparation subsystem 200, as shown in FIG. 3. In general, the media preparation subsystem 200 prepares a substrate to feed the inoculated biological material. For example, the media may include sugar, yeast, sulfuric acid, chlorine, or other suitable feed materials. The media may be prepared in a controlled atmosphere, such as under ISO 8 / Grade C / Class 100,000 conditions.

[0050]

[0059] In some embodiments, the media preparation subsystem 200 includes a manual assembly of components. In some embodiments, the components may be refrigerated within the media preparation subsystem 200. The prepared media may be refrigerated after mixing for a longer shelf life.

[0051]

[0060] The medium can be prepared according to the requirements of any given application. The medium can be a defined medium, a synthetic medium, a non-defined medium, or a basal medium. The medium can be an enriched medium or a highly enriched medium. The medium can include various amino acids, such as non-essential and essential amino acids, such as glutamine and branched chain amino acids. The concentration of the medium can be altered to manipulate the response of the cell culture of interest.

[0052]

[0061] The medium components include, for example, a buffer, an amino acid content, a vitamin content, a salt content, a mineral content, a serum content, a carbon source content, a lipid content, a nucleic acid content, a hormone content, a trace element content, an ammonia content, a cofactor content, an indicator content, a small molecule content, a hydrolysate content, and an enzyme regulator content.

[0053]

[0062] Minerals that are optionally present include bismuth, boron, calcium, chlorine, chromium, cobalt, copper, fluorine, iodine, iron, magnesium, manganese, molybdenum, nickel, phosphorus, potassium, rubidium, selenium, silicon, sodium, strontium, sulfur, tellurium, titanium, tungsten, vanadium, and zinc. Exemplary salts and minerals include CaCl2 (anhydrous), CuSO4 5H2O, Fe(NO3).9H2O, KCl, KNO3, KH2PO4, MgSO4 (anhydrous), NaCl, NaH2PO4H2O, NaHCO3, Na2SE3 (anhydrous), ZnSO4.7H2O; linoleic acid, lipoic acid, D-glucose, hypoxanthine disodium, phenol red, putrescine diHCl, sodium pyruvate, thymidine, pyruvic acid, sodium succinate, succinic acid, sodium succinate hexahydrate, glutathione (reduced), para-aminobenzoic acid (PABA), methyl linoleic acid, bactopeptone G, adenosine, cytidine, guanosine, 2'-deoxyadenosine HCl, 2'-deoxycytidine HCl, 2'-deoxyguanosine, and uridine.

[0054]

[0063] Suitable media and culture methods, for example for mammalian cell lines, are well known in the art, as described, for example, in U.S. Patent No. 5,633,162. Examples of standard cell culture media for laboratory flask or low density cell culture and adapted to the needs of a particular cell type include, for example, Roswell Park Memorial Institute (RPMI) 1640 medium (Morre, G., The Journal of the American Medical Association, 199, p. 519f. 1967), L-15 medium (Leibovitz, A. et al., Amer. J. of Hygiene, 78, 1 p. 173ff. 1963), Dulbecco's modified Eagle's Medium (DMEM), Eagle's minimal essential medium (MEM), and others. Medium), Ham's F12 medium (Ham, R. et al., Proc. Natl. Acad. Sc. 53, p288ff. 1965), or Iscoves' modified DMEM lacking albumin, transferrin, and lecithin (Iscoves et al., J. Exp. med. 1, p. 923ff., 1978). It is known that such culture media can be supplemented with fetal bovine serum (FBS, also called FCS), which provides a natural source of large amounts of hormones and growth factors. Cell culture of vertebrate and mammalian cells, respectively, is performed according to conventional methods and is covered in detail in, for example, R. Ian Fresney, Culture of Animal Cells, a manual, 4th edition, Wiley-Liss / New York, 2000.

[0055]

[0064] In some embodiments, a high density growth culture medium may be used. High density growth media can be supplemented with nutrients such as total amino acids, energy sources such as glucose, inorganic salts, vitamins, trace elements (defined as inorganic compounds that are usually present at final concentrations in the micromolar range), buffers, the four nucleosides or their corresponding nucleotides, antioxidants such as glutathione (reduced form), vitamin C, and other components such as important membrane lipids, e.g., cholesterol or phosphatidylcholine, or lipid precursors, e.g., choline or inositol. High density media are enriched in most or all of these compounds, and contain them in higher amounts (fortified) than the aforementioned standard media, such as those that can come from EP-435 911 A or GB-2251249 compared to RPMI1640, with the exception of inorganic salts, and the osmolality of the essentially isobaric medium is adjusted based on the inorganic salts. GB-2251249 provides examples of suitable high density growth media. In some embodiments, the high density cell culture medium contains acetate in the amount mentioned above, possibly in the absence of butyrate. In addition, the culture medium may be balanced and enriched in that most of the amino acids other than tryptophan exceed 75 mg / L in the culture medium. In conjunction with the general amino acid requirements, the combined amount of glutamine and asparagine may exceed 1 g / L in total, such as exceeding 2 g / L in the high density culture medium. It goes without saying that the latter more preferred embodiment is less suitable for recombinant cell lines transfected with glutamine synthetase (GS) vectors, especially after a series of amplifications of the GS gene sequence have occurred. In those cells, for example, an excess of glutamine combined from exogenous and endogenous sources leads to the production of ammonia, which should be avoided.

[0056]

[0065] Small volumes of media may be prepared in a biosafety cabinet. ISO 5 / Grade A / Class 100 controlled biosafety cabinets provide isolation for the preparation of small inoculum media.

[0057]

[0066] Generally, however, large volumes of medium can be prepared continuously or batch by batch in the SUM. For example, batches can be prepared in several SUMs in a subsystem, such as a small SUM 204, or a large SUM 206, or both, as shown in FIG. 3. The SUMs used to prepare the medium can be larger than about 100 L, such as more than about 500 L, such as more than about 1000 L, such as more than about 2000 L. Generally, however, the SUMs are smaller than about 4000 L, such as less than about 3000 L, such as less than about 2000 L.

[0058]

[0067] The buffer solutions used in the media preparation subsystem 200 may be pumped directly into the SUMs from the buffer mixture distribution manifold. Alternatively, the buffer solutions may be pumped into a temporary holding tank (e.g., tote 202) from which the buffer mixture may be dispensed into the SUMs as needed. Depending on the volume of the SUMs and / or process demands, different concentrations of buffer may be dispensed into different SUMs than are received from the first and second buffer mixture distribution manifolds.

[0059]

[0068] The prepared medium may be stored in at least one holding tank 208. In some embodiments, the prepared medium may be stored directly in a disposable bag. If necessary, the bag may be stored in a refrigerated room to increase the shelf life of the medium. In some embodiments, the holding tank is directly refrigerated.

[0060]

[0069] In some embodiments, a system provided in accordance with the present disclosure includes a bioreactor subsystem 300, as shown in Figure 4. The bioreactor subsystem may be housed in a single room that is optionally designated a controlled, non-sensitive area.

[0061]

[0070] The bioreactor subsystem 300 generally cultivates a biologic through at least one seed train prior to placement into at least one production bioreactor. For example, some embodiments initiate a seed train from a small source, such as a flask or vial, that is manually prepared in the glove box 302. For example, a flask containing an amount of inoculum medium may be shaken by hand to initiate the cultivation process in an isolated incubator. In other examples, the process is automated by pumping a small amount of medium into a flask that is pre-filled with the inoculum.

[0062]

[0071] After inoculation of the small sample, some embodiments transport the growing cell mass to several incubation stages in a seed train. For example, the seed train may proceed from a wave incubator 304 (e.g., a perfusion-based wave incubator) to a first disposable bioreactor (SUB) 306 (e.g., 20 L-500 L), to a second SUB 308 (e.g., 100 L-1000 L), to a third SUB 310 (e.g., 500 L-4000 L) before reaching a production bioreactor 312.

[0063]

[0072] The seed train may proceed by manually transferring active cell mass from one incubation stage to the next, in some embodiments, the transfer may be automated and controlled by feedback from sensors measuring cell mass, cell concentration, glucose levels, or other such progress indicators.

[0064]

[0073] In one embodiment, the seed train includes a perfusion-based reactor system 316 that feeds directly into at least one production bioreactor. For example, a perfusion reactor may reduce the time required to grow a cell culture to the size or mass required by the production bioreactor. The perfusion seed train continuously provides fresh medium to the cell culture while removing any spent medium to keep the cell culture in either a filtered section or a holding vessel (e.g., a bioreactor). In this manner, the culture is always provided with optimal growth conditions, allowing for very high culture concentrations. The perfusion-based reactor system 316 may replace one or more intermediate SUB stages in the seed train. For example, in some cases, the perfusion-based reactor system may feed directly into at least one production bioreactor, such as two bioreactors, such as three bioreactors, such as five bioreactors, etc. In some cases, all bioreactors in the bioreactor subsystem 300 may be fed by the perfusion-based reactor system 316. Media and / or buffers may be pumped directly from the media preparation subsystem 200 or the buffer distribution subsystem 100 to the perfusion reaction system 316.

[0065]

[0074] When used in conjunction with a perfusion reactor, the bioreactor 312 may or may not be specifically designed for perfusion feeding. For example, with respect to the harvest port and piping design of a bioreactor used for a perfusion mode of operation, the dip tube needs to be appropriately sized to allow unimpeded flow of culture out of the bioreactor and into the coupled cell retention device, and to be able to flow from the cell retention device back into the bioreactor without foaming and shearing of the cells or cell aggregates. The smooth flow of cell culture during the perfusion mode or harvest phase of a fed-batch process is to ensure that the cells are not mechanically damaged while passing through such piping, as mechanical damage can result in the release of cellular factors (e.g., enzymes such as glutathione reductase, thioredoxin reductase or metabolites such as thioredoxin and NADPH) that can adversely affect the performance of the process and the quality of the product made. Secondly, the smooth flow of the cell culture may result in the culture not becoming hypoxic or anoxic while passing through such tubing, thereby preventing activation of released cellular factors that may adversely affect process performance and product quality during further processing.

[0066]

[0075] The production bioreactor 312 may include stirred, rocking, airlift, or other bioreactor types. In some embodiments, the production bioreactor is a continuous, fed-batch, and / or perfusion-fed SUB. Additionally or alternatively, one or more of the production bioreactors 312 may be perfusion reactors. In some embodiments, the bioreactors may be of various diameters, such as those described in U.S. Patent Publication No. 2017 / 0369828 A1, which is incorporated herein by reference in its entirety.

[0067]

[0076] When more than one production bioreactor 312 is present in the bioreactor subsystem 300, the production bioreactors 312 can be the same or different sizes, such as greater than about 500 L, such as greater than about 1000 L, such as greater than about 2000 L, such as greater than about 5000 L, such as greater than about 8000 L, such as greater than about 12000 L, such as greater than about 18000 L, etc. In one embodiment, the production bioreactor 312 is smaller than about 20000 L, such as less than about 15000 L, such as less than about 10000 L.

[0068]

[0077] The bioreactor unit may perform one or more or all of the following: supplying nutrients and / or carbon sources, injecting a suitable gas (e.g., oxygen), inflowing fermentation or cell culture medium, separating gas and liquid phases, maintaining growth temperature, maintaining pH levels, agitation (e.g., stirring), and / or cleaning / sterilization. In one embodiment, the bioreactor is suitable for culturing suspension or attachment-dependent (adherent) cells.

[0069]

[0078] In one embodiment, the bioreactor subsystem 300 is suitable for cell therapy and / or viral therapy operations. In one embodiment, the bioreactor subsystem 300 is suitable for culturing prokaryotic or eukaryotic cells. Examples of cells include, but are not limited to, bacterial cells (e.g., E. coli, P. pastoris), yeast cells (e.g., S. cerevisae, T. Reesei), plant cells, insect cells (e.g., Sf9), Chinese Hamster Ovary cells (CHO and any genetically modified or derived CHO cell lines), mouse cells (e.g., mouse embryonic fibroblasts, cells derived from mouse cancer models), human cells (e.g., cells from any tissue or organ, cells from cancer or other diseased cell lines, stem cells), hybridoma cells, or other genetically modified or hybrid cells. In one embodiment, the cells express or produce a product, such as a recombinant therapeutic or diagnostic agent. Examples of products produced by the cells include, but are not limited to, antibody molecules (e.g., monoclonal antibodies, bispecific antibodies), fusion proteins (e.g., Fc fusion proteins, chimeric cytokines), other recombinant proteins (e.g., glycosylated proteins, enzymes, hormones), or lipid-encased particles (e.g., exosomes, virus-like particles).

[0070]

[0079] The production bioreactor 312 may optionally be installed within a bioreactor station 314. For example, the bioreactor station 314 may provide physical floor, wall, or cabinet space within a room to place the bioreactor, as well as connections for any electrical, mechanical, heating / cooling water, media, buffers, sterile WFI, and / or cell culture conduits required by the bioreactor. For example, each bioreactor station 314 may be in communication with at least one buffer distribution manifold 116 or 120 to provide the required concentrations of buffers to the bioreactor.

[0071]

[0080] Additionally or alternatively, the bioreactor station 314 may provide a connection for the output of any effluent from the bioreactor. For example, the bioreactor station 314 may be in fluid communication with a flow control assembly. When not loaded with bioreactors, fluid communication may be controlled, for example, by closed valves. However, any existing fluid communication conduits aid in the rapid expansion of bioreactor capacity by allowing new bioreactors installed at station 314 to be easily connected to the flow control assembly without the need to install additional conduits or other piping with the addition of each new bioreactor.

[0072]

[0081] In some examples, the bioreactor stations 314 are uniquely configured for a particular size, type, or configuration of bioreactor. In other examples, each bioreactor station 314 can accommodate a wide variety of bioreactors with little or no customization. In this manner, bioreactor reconfiguration is streamlined. For example, the bioreactor subsystem 300 can be quickly scaled in response to growing product demands.

[0073]

[0082] In some examples, a production bioreactor 312 is placed in each bioreactor station 314. In other examples, such as those shown in Figure 4, some bioreactor stations 314 do not include a production bioreactor 312. In such examples, an empty station waits to receive a production bioreactor if additional output is required.

[0074]

[0083] The stations 314 may be filled all at once, or may be filled in stages as desired. For example, the bioreactor subsystem may be operated once with a first portion of the stations filled. A second time, a second portion of the stations may be filled in addition to or instead of the first portion. That is, the first and second portions need not overlap for any number of the stations. The bioreactor subsystem 300 may alternate between operation on the first portion and operation on the second portion to minimize downtime between bioreactor batches.

[0075]

[0084] In some examples, a second portion of stations may be completely filled, in addition to the first portion, with this second portion being equal to or less than the amount of previously unused bioreactor stations 314. For example, a bioreactor subsystem 300 having six bioreactor stations 314 that initially operates with a first portion of four bioreactors 312 may add a second portion of two bioreactors 312 if needed. In another example, when titers fall below a design threshold, additional bioreactors may be quickly brought online to increase the total bioreactor volume. In this manner, a high-output bioreactor subsystem 300 may be built and implemented in stages as demand rises from small to large scale.

[0076]

[0085] The bioreactor subsystem 300 may be equipped with more than one bioreactor station, such as more than two, such as more than five, such as more than ten. In one embodiment, the bioreactor subsystem 300 is equipped with fewer than 30 stations, such as fewer than 20, such as fewer than ten. In some embodiments, the bioreactor subsystem 300 may be operated with less than 80% of the stations filled, such as less than 20%, such as less than 40%, such as less than 60%. In additional examples, the bioreactor subsystem may be operated with more than 20% of the stations filled, such as more than 40%, such as more than 60%, such as more than 80%, or even up to 100%. The stations may be filled or deactivated in any number of phases or steps, such as 1, 2, 3, 4, or more.

[0077]

[0086] In some embodiments, the production bioreactor may receive medium directly from the medium holding tank 208. For example, the medium may be transferred to all of the tanks containing the production bioreactor 312. The flow rate may be distributed to the bioreactor station 314 via a main line to the bioreactor subsystem 300 and then distributed to each station via a manifold or via direct lines. The flow rate in each line may be monitored and / or controlled to ensure optimal operation.

[0078]

[0087] In other examples, medium may be manually distributed to each bioreactor station 314 in a mobile container. For example, totes or bags may be filled directly in the medium preparation subsystem 200 and transferred to the bioreactor subsystem 300. In some embodiments, the medium preparation subsystem 200 and the bioreactor subsystem 300 are adjacent and a pass-through airlock may allow for easy transfer. The bioreactor station may include a bin or container for the medium-containing totes or bags. In such examples, once the bin or container is filled, the flow of medium into the bioreactor may be monitored and / or controlled to ensure optimal operation.

[0079]

[0088] For example, control may follow an open loop (e.g., a timer) or a closed loop (e.g., in response to feedback from sensors reading temperature, cell density, glucose levels, etc.). The control system may also notify the operator of the need to refill media containers of the bioreactor station 314 in some examples.

[0080]

[0089] Systems prepared according to the present disclosure may, in some embodiments, include at least one harvest subsystem 400. For example, as shown in FIG. 5, the system may include two harvest subsystems 400, each having a centrifuge 402 that may receive the effluent of the bioreactor subsystem 300. For example, the centrifuge 402 may remove cells and large cellular solids from the bioreactor effluent. The output of the centrifuge 402 may be passed to a series of one or more filtration steps. For example, a bank of depth filters 404 may further purify the harvest stream and reduce turbidity to a target level. In addition, the depth filters 404 may be charged to remove other impurities, such as removing endotoxins, viral particles, and / or protein impurities.

[0081]

[0090] After at least one centrifugation, clarification, and / or filtration step, the harvest flow may be directed to at least one harvest tank 406. In some examples, each harvest subsystem 400 includes at least one harvest tank 406. In other examples, two or more harvest subsystems may share at least one harvest tank 406. For example, multiple centrifuges 402 and multiple depth filters 404 may optionally pass effluent to one tank 406. The harvest tank 406 may be connected to at least one buffer mixture distribution manifold 116 or 120 for controlling the pH of the harvest solution. In some examples, multiple harvest subsystems 400, each configured independently, may operate in parallel and may be located adjacent to one another or may be removed from one another.

[0082]

[0091] In some embodiments, a system provided in accordance with the present disclosure includes a purification subsystem 500, as shown in the exemplary embodiment of Figure 5. In some embodiments, the purification subsystem 500 can be divided into a pre-viral train 500-1 and a post-viral train 500-2.

[0083]

[0092] Pre-viral train 500-1 may include at least one chromatography, filtration, or mixing device.

[0093] One-dimensional (1D) chromatography suitable for use in the methods described herein are known to those of skill in the art and include, for example, affinity chromatography, gel filtration chromatography, ion exchange chromatography, reversed phase chromatography, and hydrophobic interaction chromatography. In some embodiments, the 1D chromatographic method is HPLC reversed phase chromatography. Chromatography. Chromatography can include high performance liquid chromatography (HPLC), gas chromatography (GC), capillary electrophoresis, ion mobility. See also, for example, Process Scale Purification of Antibodies, Uwe Gottschalk 2011 John Wiley & Sons ISBN:1118210743; Antibodies Vol1 Production and Purification, G.Subramanian 2013 Springer Science & Business Media; Basic Methods in Antibody Production and Characterization, Gary C.Howard 2000 CRC Press.

[0084]

[0094] Additional exemplary chromatographic methods include, but are not limited to, strong anion chromatography (SAX), liquid chromatography (LC), high performance liquid chromatography (HPLC), ultra performance liquid chromatography (UPLC), thin layer chromatography (TLC), amide column chromatography, and combinations thereof. Exemplary mass spectrometry (MS) includes, but is not limited to, tandem MS, LC-MS, LC-MS / MS, matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS), Fourier transform mass spectrometry (FTMS), ion mobility separation mass spectrometry (IMS-MS), electron transfer dissociation (ETD-MS), and combinations thereof. Exemplary electrophoretic methods include, but are not limited to, capillary electrophoresis (CE), CE-MS, gel electrophoresis, agarose gel electrophoresis, acrylamide gel electrophoresis, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting using antibodies that recognize specific glycan structures, and combinations thereof. Exemplary nuclear magnetic resonance (NMR) methods include, but are not limited to, one-dimensional NMR (1D-NMR), two-dimensional NMR (2D-NMR), correlation spectroscopy magnetic angle spinning NMR (COSY-NMR), total correlation spectroscopy NMR (TOCSY-NMR), heteronuclear single quantum coherence NMR (HSQC-NMR), heteronuclear multiple quantum coherence (HMQC-NMR), rotating nuclear Overhauser effect spectroscopy NMR (ROESY-NMR), nuclear Overhauser effect spectroscopy (NOESY-NMR), and combinations thereof.

[0085]

[0095] Suitable spectroscopic techniques include, for example, Raman spectroscopy. Raman spectroscopy is a technique that is gaining popularity in different specialties of forensic science. Some examples of its use today are in the detection of drugs (Hodges et al., "The Use of Fourier Transformation"). Transform Raman Spectroscopy in the Forensic Identification of Illicit Drugs and Explosives”, Molecular Spectroscopy 46:303-307 (1990)), lipstick (Rodger et al., “The In-Situ Analysis of Lipsticks by Surface Enhanced Resonance Raman Scattering”, Analyst 1823-1826 (1998)), and fibers (Thomas et al., “Raman Spectroscopy and the Forensic Analysis of Black / Grey and Blue Cotton Fibres Part1: Investigation of the Effects of Varying Laser Wavelength”, Forensic Sci.Int.152:189-197(2005)), and paint (Suzuki et al., “In Situ Identification and Analysis of Automotive Paint Pigments Using Line Segment Excitation Raman Spectroscopy: I. Inorganic Topcoat Pigments”, J.Forensic Sci.46:1 053-1069 (2001)), and ink (Mazzella et al., "Raman Spectroscopy of Blue Gel Pen Inks," Forensic Sci. Int. 152:241-247 (2005)) analysis. The theory behind Raman spectroscopy is based on the inelastic scattering of low intensity nondestructive laser light by a solid, liquid, or gas sample. Little or no sample preparation is required, and the amount of test material required is a few picograms or femtoliters (10 -12 Grams or 10 -15The amount of Raman spectroscopy can be as little as 1 liter. A typical Raman spectrum consists of several narrow bands, providing the unique vibrational signature of the material (Grasselli et al., "Chemical Applications of Raman Spectroscopy", New York: John Wiley & Sons (1981)). Unlike another type of vibrational spectroscopy, infrared (IR) absorption spectroscopy, Raman spectroscopy shows very little interference from water (Grasselli et al., "Chemical Applications of Raman Spectroscopy", New York: John Wiley & Sons (1981)), making it an excellent technique for analyzing body fluids and their traces. Proper Raman spectroscopy does not damage the sample. Stain or swab can be field tested and still be available for further use in the laboratory for DNA analysis, which is very important for forensic applications. The design of portable Raman spectrometers that provide crime scene identification capabilities is now a reality (Yan et al., "Surface-Enhanced Raman Scattering Detection of Chemical and Biological Agents Using a Portable Raman Integrated Tunable Sensor", Sensors and Actuators B.6 (2007); Eckenrode et al., "Portable Raman Spectroscopy systems for Field Analysis", Forensic Science Communications 3:(2001)).

[0086]

[0096] Types of Raman spectroscopy suitable for use in conjunction with the present disclosure include, but are not limited to, conventional Raman spectroscopy, Raman microspectroscopy, near-field Raman spectroscopy including but not limited to tip-enhanced Raman spectroscopy, surface-enhanced Raman spectroscopy (SERS) and surface-enhanced resonance Raman spectroscopy (SERRS), coherent anti-Stokes Raman spectroscopy (CARS), etc. Both Stokes and anti-Stokes Raman spectroscopy may be used.

[0087]

[0097] In some embodiments, rapid non-invasive testing is performed on one or more bioprocess intermediates using chromatographic or spectroscopic methods described herein, hi some embodiments, rapid testing is performed using Raman spectroscopy.

[0088]

[0098] 6, the harvest stream from the harvest tank 406 is delivered to a chromatography skid 504 and its associated chromatography column 502. After the first chromatography step, the product stream may be passed to a SUM 506, such as for viral inactivation, before proceeding to a filtration device 508.

[0089]

[0099] The filtration device 508 may include various filtration mechanisms. In some examples, a tangential flow filtration (TFF) device is used. For example, the TFF device may allow for diafiltration of the product stream. In such examples, the TFF device may optionally be connected to at least one buffer mixture distribution manifold 116 or 120 for buffer exchange or concentration manipulation. The TFF may optionally work together with the SUM 506 for retentate recirculation and / or retention.

[0090]

[0100] In some embodiments, the TFF process consists of two stages: volume reduction and diafiltration. During the volume reduction step, the bulk volume (e.g., cell culture medium) is filtered into the pass-through side of the filter until the desired product concentration is reached in the processing bag or holding tank. In the diafiltration stage after the volume reduction stage, the concentrated product is washed with a fluid such as a buffer to remove undesirable or unacceptable cell culture or harvest medium components. Further volume reduction may also be performed after diafiltration to reach the desired product density.

[0091]

[0101] The pre-viral train 500-1 may include any number of chromatography, filtration, mixing, and / or other desired devices. For example, FIG. 6 shows a pre-viral train with three chromatography stages. However, the chromatography, filtration, and / or mixing devices may each be present in any quantity, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, or even 20 or 30, with each device present in an independently selected amount.

[0092]

[0102] The post-viral train 500-2 may include any number of desired finishing devices. For example, the post-viral train may include one or more final polishing chromatography and / or filtration stages in addition to or instead of the chromatography stages in the pre-viral train 500-1. The critical virus reduction step is considered to be a suitable point for spatial isolation, and the post-viral separation subunit is considered to be essentially virus-free. The post-viral separation subunit houses equipment and utilities suitable for any one of ultrafiltration (tangential filtration), regular filtration, chromatography, formulation, titration, mixing, concentration, buffer exchange, bulk drug substance container filling, and refrigeration.

[0093]

[0103] In some examples, the post virus train may include empty equipment stations, such as a chromatography column station 512, a chromatography skid station 514, a SUM station 516, and / or a TFF device station 518, as shown in Figure 6. Similar to the bioreactor station 314, the various stations 512, 514, and 516 may provide the respective devices with the required electrical and mechanical connections for operation, as well as any necessary heated / cooled water, sterile WFI, product streams, buffers, media, filtration media, sorbents, eluents, or any other inflows required by the respective devices. For example, the chromatography column station 512 may provide the necessary electrical, sorbent, eluent, and / or product stream connections while also providing physical floor, wall, or cabinet space for mounting.

[0094]

[0104] Similar to that discussed with respect to bioreactor station 314, the chromatography columns and skid stations may be present in filled, empty, or inactivated volumes, in any of the exemplary quantities described above, or in other quantities as desired.

[0095]

[0105] Optionally, both the pre-virus train and the post-virus train may include equipment installed in associated stations. For example, each chromatography column and skid in pre-virus train 500-1 may be installed in column station 512 and skid station 514, respectively. If the post-virus train also has at least one column station 512 and skid station 514, any of the chromatography columns or skids in pre-virus train 500-1 may be easily removed from pre-virus train 500-1 and installed in the post-virus train. Train 500-2 can be quickly installed. Optionally, both the pre-viral and post-viral trains can include at least one empty station to allow for easy and rapid insertion of additional equipment if the need arises. For example, all stations required to accommodate a parallel pre-viral train can be housed in pre-viral train 500-1 for future throughput requirements. In this manner, the purification subsystem can be quickly and flexibly adapted for incremental growth.

[0096]

[0106] The post-virus train 500-2 may further include several different filling devices 520. For example, the filling device 520 may be a bottle filling device, or alternatively a bag filling device, or generally a filling device container. The device may in some examples be a self-contained unit, optionally having its own controlled airspace, such as an airspace controlled to ISO 5 / Grade A / Class 100.

[0097]

[0107] In some embodiments, the maintenance area 500-3 may be adjacent to one or both of the pre-virus and post-virus trains. When adjacent to both, airlocks 510 allow access from each train as well as from outside the purification subsystem 500. For example, the maintenance area 500-3 may be used for equipment cleaning, repair storage, or staging. For example, chromatography columns may be packaged and prepared for rapid insertion into either the pre-virus or post-virus trains. In some embodiments, the maintenance area 500-3 has both a chromatography column station 512 and a chromatography skid station 514.

[0098]

[0108] The description of various embodiments of the present disclosure may be utilized for the production of pharmaceutical and biopharmaceutical products. The devices, facilities, and methods described herein are suitable for culturing any desired cell line, including prokaryotic and / or eukaryotic cell lines. Moreover, in embodiments, the devices, facilities, and methods are suitable for culturing suspension cells or anchorage-dependent (adherent) cells, and are suitable for production operations configured for the production of pharmaceutical and biopharmaceutical products, such as polypeptide products, nucleic acid products (e.g., DNA or RNA), or cells and / or viruses, such as those used in cell therapy and / or virus therapy.

[0099]

[0109] In embodiments, the cells express or produce a product, such as a recombinant therapeutic or diagnostic agent. As described in more detail below, examples of products produced by the cells include, but are not limited to, 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), 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), cellular therapeutics (e.g., pluripotent stem cells, mesenchymal stem cells, and adult stem cells), vaccines or lipid-coated particles (e.g., exosomes, virus-like particles), RNA (e.g., siRNA) or DNA (e.g., plasmid DNA), antibiotics, or amino acids. In embodiments, the devices, facilities, and methods can be used to produce biosimilars.

[0100]

[0110] As mentioned, in embodiments, the devices, facilities, and methods involve the use of eukaryotic cells, e.g., mammalian cells or lower eukaryotic cells, e.g., yeast cells or filamentous fungal cells, or prokaryotic cells, such as gram-positive or gram-negative cells, and / or products of eukaryotic or prokaryotic cells, e.g., proteins, which are synthesized by eukaryotic cells in a large-scale manner. , peptides, antibiotics, amino acids, nucleic acids (such as DNA or RNA). Unless otherwise stated herein, the devices, facilities, and methods may include any desired volume or production capacity, including, but not limited to, bench scale, pilot scale, full production scale capacity.

[0101]

[0111] In an embodiment, the cell is a eukaryotic cell (e.g., a mammalian cell). The mammalian cell may be, for example, a human or murine or bovine cell line or cell line. Such cells, cell lines or cell lines are, for example, mouse myeloma (NSO) cell lines, Chinese hamster ovary (CHO) cell lines, HT1080, H9, HepG2, MCF7, MDBK Jurkat, NIH3T3, PC12, BHK (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. Preferably, the mammalian cell is a CHO cell line. In one embodiment, the cell is a CHO cell. In one embodiment, the cell is a CHO-K1 cell, a CHO-K1 SV cell, a DG44 CHO cell, a DUXB11 CHO cell, a CHOS, a CHO GS knockout cell, a CHO FUT8 GS knockout cell, a CHOZN, or a CHO-derived cell. The CHO GS knockout cell (e.g., a GSKO cell) is, for example, a CHO-K1 SV GS knockout cell. The CHO FUT8 knockout cell is, for example, Potelligent® CHOK1 SV (Lonza Biologics, Inc.). The eukaryotic cell may also be an avian cell, cell line, or cell line, such as, for example, an EBx® cell, EB14, EB24, EB26, EB66, or EBv13.

[0102]

[0112] In one embodiment, the eukaryotic cell is a stem cell.The stem cell may be a pluripotent stem cell, including, for example, embryonic stem cell (ESC), adult stem cell, induced pluripotent stem cell (iPSC), tissue-specific stem cell (e.g., hematopoietic stem cell), and mesenchymal stem cell (MSC).

[0103]

[0113] In one embodiment, the cells are any differentiated form of the cells described herein, hi one embodiment, the cells are derived from any progenitor cell in culture.

[0104]

[0114] In an embodiment, the cell is a hepatocyte, such as a human hepatocyte, an animal hepatocyte, or a non-parenchymal cell. For example, the cell may be an adherent metabolic test human hepatocyte, an adherent induction test human hepatocyte, an adherent Qualyst Transporter Certified™ human hepatocyte, a suspension test human hepatocyte (including 10 donor and 20 donor pooled hepatocytes), a human hepatic Kupffer cell, a human hepatic stellate cell, a dog hepatocyte (including single and pooled beagle hepatocytes), a mouse hepatocyte (including CD-1 and C57BI / 6 hepatocytes), a rat hepatocyte (including Sprague-Dawley, Wistar Han, and Wistar hepatocytes), a monkey hepatocyte (including cynomolgus or rhesus hepatocytes), a cat hepatocyte (domestic shorthair hepatocyte), and a rabbit hepatocyte (New Zealand white hepatocyte). Exemplary hepatocytes are commercially available from Triangle Research Labs, LLC, 6 Davis Drive Research Triangle Park, NC, USA 27709.

[0105]

[0115] In one embodiment, the eukaryotic cell is a yeast cell (e.g., a cell of the genus Pichia (e.g., Pichia pastoris, Pichia methanolica, Pichia kluyveri, and Pichia angusta), a cell of the genus Komagataella (e.g., Komagataella pastoris, Komagataella spp. ... magataella pseudopastoris, or Komagataella phaffii), Saccharomyces (e.g., Saccharomyces cerevisae, cerevisiae, Saccharomyces kluyveri, Saccharomyces uvarum), Kluyveromyces (e.g., Kluyveromyces lactis, Kluyveromyces marxianus), Candida (e.g., Candida utilis, Candida cacaoi, Candida boidinii), Geotrichum (e.g., Geotrichum fermentans), Hansenula polymorpha, Yarrowia lipolytica, or Schizosaccharomyces pombe. Preferred is the species Pichia pastoris. Examples of Pichia pastoris strains are X33, GS115, KM71, KM71H, and CBS7435.

[0106]

[0116] In one embodiment, the eukaryotic cell is a fungal cell (e.g., a cell of the genus Aspergillus (e.g., A. niger, A. fumigatus, A. orzyae, A. nidula, etc.), Acremonium (e.g., A. thermophilum, etc.), Chaetomium (e.g., C. thermophilum, etc.), Chrysosporium (e.g., C. thermophile, etc.), Cordyceps (e.g., C. militaris, etc.), Corynascus, Cytenomyces, Fusarium (e.g., F. oxysporum, etc.), Glomerella (e.g., G. graminicola, etc.), etc.), Hypocrea (e.g. H. jecorina), Magnaporthe (e.g. M. orzyae), Myceliophthora (e.g. M. thermophile), Nectria (e.g. N. heamatococca), Neurospora (e.g. N. crassa), Penicillium, Sporotrichum (e.g. S. thermophile), Thielavia (e.g. T. terrestris, T. heterothallica), Trichoderma (e.g. T. reesei), or Verticillium (e.g. V. dahlia).

[0107]

[0117] In one embodiment, the eukaryotic cell is an insect cell (e.g., an Sf9, Mimic™ Sf9, Sf21, High Five™ (BT1-TN-5B1-4), or T1-Ea88 cell), an algae cell (e.g., of the genus Amphora, Bacillariophyceae, Dunaliella, Chlorella, Chlamydomonas, Cyanobacteria, Nannochloropsis, Spirulina, or Ochromonas), or a plant cell (e.g., a monocotyledonous (e.g., maize, rice, wheat, or Setaria) or dicotyledonous (e.g., cassava, potato, soybean, tomato, tobacco, alfalfa, Physcomitrella patens, or Arabidopsis cell).

[0108]

[0118] In one embodiment, the cell is a bacterial or prokaryotic cell.

[0119] In an embodiment, the prokaryotic cell is a gram-positive cell, such as bacillus, streptomyces, streptococcus, staphylococcus, or lactobacillus.The bacillus that can be used is, for example, B.subtilis, B.amyloliquefaciens, B.licheniformis, B.natto, or B.megaterium.In an embodiment, the cell is B.subtilis, such as B.subtilis 3NA and B.subtilis 168.The bacillus can be obtained, for example, from Bacillus Genetic Stock Center, Biological Sciences 556, 484 West12th Avenue, Columbus Ohio 43210-1214.

[0109]

[0120] In one embodiment, the prokaryotic cell is a Salmonella spp., or Escherichia coli, e.g., TG1, TG2, W3110, DH1, DHB4, DH5a, HMS174, HMS1 74(DE3), NM533, C600, HB101, JM109, MC4100, XL1-Blue, and Origami, as well as gram-negative cells such as those derived from E. coli B strains, e.g., BL-21 or BL21(DE3), all of which are commercially available.

[0110]

[0121] Suitable host cells are commercially available from microorganism collections such as, for example, DSMZ (Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH, Braunschweig, Germany) or the American Type Culture Collection (ATCC).

[0111]

[0122] In embodiments, the cultured cells are used to produce proteins, e.g., antibodies, e.g., monoclonal antibodies, and / or recombinant proteins, for therapeutic use. In embodiments, the cultured cells produce peptides, amino acids, fatty acids, or other useful biochemical intermediates or metabolites. For example, in embodiments, molecules having molecular weights from about 4000 daltons to greater than about 140,000 daltons can be produced. In embodiments, these molecules can have a range of complexity and can include post-translational modifications, including glycosylation.

[0112]

[0123] In embodiments, the protein is, for example, BOTOX, Myobloc, Neurobloc, Dysport (or other serotypes of botulinum neurotoxin), alglucosidase alfa, daptomycin, YH-16, chorionic gonadotropin alfa, filgrastim, cetrorelix, interleukin-2, aldesleukin, teceleulin, denileukin diftitox, interferon alfa-n3 (injection), interferon alfa-n1, DL-8234, interferon, Suntory (gamma -1a), Interferon gamma, Thymosin alpha 1, Tasonermin, DigiFab, ViperaTAb, EchiTAb, CroFab, Nesiritide, Abatacept, Alefacept, Rebif, Eptothermin alfa, Teriparatide (osteoporosis), Injectable calcitonin (bone disease), Calcitonin (nasal, osteoporosis), Etanercept, Hemoglobin glutamer 250 (bovine), Drotrecogin alfa, Collagenase, Carperitide, Recombinant human epidermal growth factor (topical gel, wound healing), DWP401, Darbepoetin alfa, Epoetin Omega, Epoetin beta, Epoetin alfa, Desirudin, Lepirudin, Bivalirudin, Nonacog alfa, Mononine, Eptacog alfa (activated), rFVIII + VWF, Reconate, rFVIII, rFVIII (recombinant), Alphnmate, Octocog alfa, rFVIII, Palifermin, Indikinase, Tenecteplase, Alteplase, Pamiteplase, Reteplase, Nateplase, Monteplase, Follitropin alfa, rFSH, hpFSH, Micafungin, Pe Gfilgrastim, Lenograstim, Nartograstim, Sermorelin, Glucagon, Exenatide, Pramlintide, Iniglucerase, Galsulfase, Leucotropin, Molgramostim, Triptorelin acetate, Histrelin (subcutaneous implant, Hydron), Deslorelin, Histrelin, Nafarelin, Leuprolide sustained release formulation (ATRIGEL), Leuprolide implant (DUROS), Goserelin, Eutropin, KP-102 program, Somatropin, Mecasermin (growth inhibition),Enlfavirtide, Org-33408, insulin glargine, insulin glulisine, insulin (inhaled), insulin lispro, insulin deternir, insulin (buccal, RapidMist), mecasermin linfavert, anakinra, cermoleukin, 99mTc-apsitide injection, myelopid, Betaseron, glatiramer acetate, Gepon, sargramostim, oprelvekin, human leukocyte-derived alpha interferon, Bilive, insulin (recombinant, e), recombinant human insulin, insulin aspart, mecasenin, Roferon-A, interferon-alpha 2, Alfaferone, interferon alfacon-1, interferon alpha, Avonex' recombinant human luteinizing hormone, dornase alfa, trafermin, ziconotide, taltirelin, divotermin alfa, atosiban, becaplermin, eptifibatide, Zemaira, CTC-111, Shanvac-B, HPV vaccine (quadrivalent), octreotide, lanreotide, and Sestim, agalsidase beta, agalsidase alpha, laronidase, copper acetate prezatide (topical gel), rasburicase, ranibizumab, Actimmune, PEG-Intron, Tricomin, recombinant house dust mite allergy desensitization injection, recombinant human parathyroid hormone (PTH) 1-84 (subcutaneous injection, osteoporosis), epoetin delta, transgenic antithrombin III, Granditropin, Vitrase, recombinant insulin, interferon-alpha (oral lozenge), GEM-21S, vapreotide, idurus lufase, omnapatrilat, recombinant serum albumin, certolizumab pegol, glucarpidase, human recombinant C1 esterase inhibitor (angioedema), lanoteplase, recombinant human growth hormone, enfuvirtide (needle-free injection, Biojector2000), VGV-1, interferon (alpha), lucinactant, aviptadil (inhalation, pulmonary disease), icatibant, ecallantide, omiganan, aurograb, pexiganan acetate, ADI-PEG-20, LDI-200, degarelix, syntredelin besed. Cintredelinbesudotox, Favld, MDX-1379, ISAtx-247, liraglutide, teriparatide (osteoporosis), tifacogin, AA4500, T4N5 liposomal cosmetic solution, catumaxomab, DWP413, ART-123, Chrysalin, desmoteplase, amediplase, corifollitropin alfa, TH-9507, teduglutide, Diamyd, DWP-412, growth hormone (sustained release injection), recombinant G-CSF, insulin (inhaled, AIR), insulin (inhaled,Technosphere), insulin (inhaled, AERx), RGN-303, DiaPep277, interferon beta (hepatitis C virus infection (HCV)), interferon alpha-n3 (oral), belatacept, transdermal insulin patch, AMG-531, MBP-8298, Xerecept, opevacan, AIDSVAX, GV-1001, LymphoScan, ranpirnase, Lipoxysan, ruspultide, MP52 (beta-tricalcium phosphate carrier, bone regeneration), melanoma vaccine, sipuleucel-T (sipuleuc el-T), CTP-37, Insegia, Vitespen, Human thrombin (freezing, surgical bleeding), Thrombin, TransMID, Alfimeprase, Puricase, Terlipressin (intravenous, hepatorenal syndrome), EUR-1008M, Recombinant FGF-I (injectable, vascular disease), BDM-E, Rotigaptide, ETC-216, P-113, MBI-594AN, Duramycin (inhaled, cystic fibrosis), SCV-07, OPI-45, Endostatin, Angiostatin, ABT-510, Bowman-Birk inhibitor concentrate, XMP-6 29, 99mTc-Hynic-Annexin V, Kahalalide F, CTCE-9908, Teverelix (sustained release), Ozarelix, Rornidepsin, BAY-504798, Interleukin 4, PRX-321, Pepscan, Ivoctadequin, rhlactoferrin, TRU-015, IL-21, ATN-161, Cilengitide, Albuferon, Biphasix, IRX-2, Omega interferon, PCK-3145, CAP-232, Pa Sileotide, huN901-DMI, ovarian cancer immunotherapy vaccine, SB-249553, Oncovax-CL, OncoVax-P, BLP-25, CerVax-16, multi-epitope peptide melanoma vaccine (MART-1, gp100, tyrosinase), nemifitide, rAAT (inhaled), rAAT (dermatology), CGRP (inhaled, asthma), pegsnercept, thymosin beta 4, plitidepsin, GTP-200, ramoplanin, GRASPA, OBI-1, AC-100, salmon calcitonin (oral, eligen), calcitonin (oral,Osteoporosis, disease), examorelin, capromorelin, Cardeva, bellafermin, 131I-TM-601, KK-220, T-10, ularitide, deperestat, hematide, Chrysalin (topical), rNAPc2, recombinant factor V111 (pegylated liposomal formulation), bFGF, pegylated recombinant staphylokinase mutant, V-10153, SonoLysis Prolyse, NeuroVax, CZEN-002, Islet Cell Neogenesis Therapy, rGLP-1, BIM-51077, LY-548806, Exenatide (Controlled Release, Medisorb), AVE-0010, GA-GCB, Avorelin, ACM-9604, Linaclotide Acetate, CETi-1, Hemospan, VAL (Injectable), Rapid Acting Insulin (Injectable, Viadel), Intranasal Insulin, Insulin (Inhaled), Insulin (Oral, Eligen) , recombinant methionyl human leptin, pitrakinra subcutaneous injection, eczema), pitrakinra (inhaled dry powder, asthma), Multikine, RG-1068, MM-093, NBI-6024, AT-001, PI-0824, Org-39141, Cpn10 (autoimmune disease / inflammation), talactoferrin (topical), rEV-131 (ophthalmology), rEV-131 (respiratory disease), oral recombinant human insulin (diabetes), RPI-78M, oprelvekin (oral), CYT-99007 CTLA4-Ig, DTY-001, Balategrast, Interferon alfa-n3 (topical), IRX-3, RDP-58, Tauferon, Bile salt-stimulated lipase, Merispase, Alarine phosphatase, EP-2104R, Melanotan-II, Bremelanotide, ATL-104, Recombinant human microplasmin, AX-200, SEMAX, ACV-1, Xen-2174, CJC-1008, Dynorphin A, SI-6603, LAB GHRH, AER-002, BGC-728, Malaria vaccine (virosome, PeviPRO), ALTU-135, Parvovirus B19 vaccine, Influenza vaccine (recombinant neuraminidase), Malaria / HBV vaccine, Anthrax vaccine, Vacc-5q, Vacc-4x, HIV vaccine (oral), HPV vaccine, TatToxoid, YSPSL, CHS-13340, PTH(1-34) liposomal cream (Novasome), Ostabolin-C, PTH analog (topical, psoriasis), MBRI-93.02, MTB72F vaccine (tuberculosis), MVA-Ag85A vaccine (tuberculosis), FARA04, BA-210, recombinant plague FIV vaccine, AG-702, OxSODrol, rBetV1, Der-p1 / Der-p2 / Der-p7 allergen targeted vaccine (house dust mite allergy), PR1 peptide antigen (leukemia), mutant ras vaccine, HPV-16 E7 lipopeptide vaccine, Labirinthin vaccine (adenocarcinoma), CML vaccine, WT1-peptide vaccine (cancer), IDD-5, CDX-110, Pentrys, Norelin, CytoFab, P-9808, VT-111, Icrocaptide, Telbermin (dermatology, diabetic foot ulcer), Rupintrivir, Reticulose, rGRF, HA, alpha-galactosidase A, ACE-011, ALTU-140, CGX-1160, Angio Tensin therapeutic vaccine, D-4F, ETC-642, APP-018, rhMBL, SCV-07 (oral, tuberculosis), DRF-7295, ABT-828, ErbB2-specific immunotoxin (anticancer), DT3SSIL-3, TST-10088, PRO-1762, Combotox, cholecystokinin-B / gastrin receptor binding peptide, 111In-hEGF, AE-37, trasnizumab-DM1, Antagonist G, IL-12 (recombinant), PM-02734, IMP-321, rhIGF-BP3, BLX-883, CUV-1647 (local), L-19-based radioimmunotherapy (cancer), Re-188-P-2045, AMG-386, DC / 1540 / KLH vaccine (cancer), VX-001, AVE-9633, AC-9301, NY-ESO-1 vaccine (peptide), NA17.A2 peptide, melanoma vaccine (pulsed antigen therapy), prostate cancer vaccine, CBP-501, recombinant human lactoferrin (Drya i), FX-06, AP-214, WAP-8294A (injectable), ACP-HIP, SUN-11031, peptide YY[3-36] (obesity, intranasal), FGLL, Atacise puto, BR3-Fc, BN-003, BA-058, human parathyroid hormone 1-34 (nose, osteoporosis), F-18-CCR1, AT-1100 (celiac disease / diabetes), JPD-003, PTH(7-34) liposomal cream (Novasome), duramycin (ophthalmology, dry eye), CAB-2, CTCE-0214, glycoPEGylated erythropoietin, EPO-Fc, CNTO-528, AMG-114, JR-013, factor XIII, aminocandin, PN-951 , 716155, SUN-E7001, TH-0318, BAY-73-7977, Teverelix (immediate release), EP-51216, hGH (controlled release, Biosphere), OGP-I, sifvirtide, TV4710, ALG-889, Org-41259, rhCC10, F-991, thymopentin (pulmonary disease), r(m)CRP, hepatoselective insulin, subarin, L19-IL-2 fusion protein, elafin, NMK-150, ALTU-139, EN-122004, rhTPO, thrombopoietin Receptor agonists (thrombocytopenic disorders), AL-108, AL-208, nerve growth factor agonists (pain), SLV-317, CGX-1007, INNO-105, oral teriparatide (eligen), GEM-OS1, AC-162352, PRX-302, LFn-p24 fusion vaccine (Therapore), EP-1043, pneumococcal pediatric vaccine, malaria vaccine, meningococcal group B vaccine, neonatal group B streptococcal vaccine, anthrax vaccine, HCV vaccine (gpE1+gpE2+MF-59), otitis media therapy , HCV vaccine (core antigen + ISCOMATRIX), hPTH(1-34) (transdermal, ViaDerm), 768974, SYN-101, PGN-0052, aviscumnine, BIM-23190, tuberculosis vaccine, multi-epitope tyrosinase peptide, cancer vaccine, encastim, APC-8024, GI-5005, ACC-001, TTS-CD3, vascular targeted TNF (solid tumors), desmopressin (buccal controlled release), onercept, and TP-9201.

[0113]

[0124] In some embodiments, the polypeptide is adalimumab (HUMIRA), infliximab (REMICADE™), rituximab (RITUXAN™ / MAB THERA™), etanercept (ENBREL™), bevacizumab (AVASTIN™), trastuzumab (HERCEPTIN™), pegrillgrastim (NEULASTA™), or any other suitable polypeptide, including biosimilars and biobetters.

[0114]

[0125] Other suitable polypeptides are those listed in Table 1 of US2016 / 0097074 and Tables 1-3 of US2017 / 0260763.

[0126] These and other modifications and variations to the present invention may be practiced by those skilled in the art without departing from the spirit and scope of the present invention, as more particularly set forth in the appended claims. In addition, it should be understood that aspects of the various embodiments may be interchanged in whole or in part. Moreover, those skilled in the art should appreciate that the foregoing description is merely exemplary and is not intended to limit the invention as further set forth in such appended claims.

Claims

1. a bioreactor subsystem comprising at least two bioreactors; at least two harvesting subsystems, each harvesting subsystem, either alone or in combination, in fluid communication with each of the bioreactors in the bioreactor subsystem; 1. A system for growing a biologic comprising:

2. 10. The system of claim 1, further comprising a flow control assembly positioned between the at least two bioreactors and the at least two harvest subsystems, the flow control assembly configured to selectively control a flow rate of fluid from a first bioreactor to a first harvest subsystem and from the first bioreactor to a second harvest subsystem, the flow control assembly further configured to control a flow rate of fluid from a second bioreactor to the first harvest subsystem and from the second bioreactor to the second harvest subsystem.

3. 10. The system of claim 1, further comprising at least one purification subsystem, each purification subsystem, either alone or in combination, in fluid communication with each of said harvesting subsystems.

4. 10. The system of claim 1, wherein at least one of the purification subsystems is configured to produce a different biologic than at least one other purification subsystem.

5. 10. The system of claim 1 configured to process more than 100 batches per year.

6. 10. The system of claim 1, wherein at least one of the bioreactors is 6000 L or larger.

7. 10. The system of claim 1, wherein the bioreactor subsystem comprises a seed train of a fed-batch culturator.

8. 10. The system of claim 1, wherein the bioreactor subsystem comprises at least one perfusion seed train.

9. 10. The system of claim 8, wherein the bioreactor subsystem comprises at least one fed-batch bioreactor.

10. At least one buffer dilution device that receives at least two inputs (i) and (ii), (i) being a buffer concentrate source stream and (ii) being sterile water for an injection source stream; at least one buffer dilution device that delivers a buffer mixture comprising (i) and (ii) to a buffer mixture distribution manifold; At least one buffer mixture distribution manifold may be used to control parameters of the buffer mixture delivered independently to each of a plurality of destinations, the parameters including at least one of flow rate or concentration; A system for dispensing buffer solutions, wherein the buffer mixture is not stored prior to delivery to any of the plurality of destinations.

11. A first buffer dilution device, wherein the buffer mixture output from the first buffer dilution device is fed as input (i) to at least one downstream buffer dilution device. a first buffer dilution device, the inflow (i) of any buffer dilution device downstream of the first buffer dilution device may be partially diverted to another buffer dilution device or to at least one buffer mixture distribution manifold; 11. The system of claim 10, wherein at least two buffer mixture distribution manifolds distribute buffers of different concentrations.

12. 11. The system of claim 10, wherein the at least two buffer dilution devices deliver approximately the same concentration of buffer to the at least one buffer distribution manifold.

13. 11. The system of claim 10, wherein at least one buffer destination comprises a media preparation subsystem, a cell culture subsystem, a bioreactor subsystem, a harvest subsystem, and / or a purification subsystem.

14. Perfused seed train; a bioreactor subsystem comprising at least one bioreactor fed in whole or in part from the perfused seed train; at least two harvesting subsystems, each harvesting subsystem, either alone or in combination, in fluid communication with each of the bioreactors in the bioreactor subsystem; 1. A system for growing a biologic comprising:

15. A building comprising the system according to claim 14.

16. Each of the at least two purification subsystems comprises: three chromatography column stations and three chromatography skid stations in the first room; a chromatography column station and a chromatography skid station in a second adjacent room; a maintenance area accessible from either the first room or the second room; 16. The building of claim 15, wherein the maintenance area is accessible from the first and second rooms by an airlock and is further accessible from the remainder of the building by another airlock.

17. 17. The building of claim 16, wherein the second room further comprises a filling device.

18. 16. The building of claim 15, wherein the bioreactor subsystem comprises multiple bioreactor stations in one room, some or all of the stations being filled with bioreactors.

19. 20. The system of claim 18, wherein at least one bioreactor station is not filled with a bioreactor.

20. 20. The system of claim 19, wherein at least one of the bioreactor stations is not filled with a bioreactor and at least one of the bioreactor stations that is filled with a bioreactor is configured to maintain fluid communication with a flow control assembly, the flow control assembly configured to selectively control a flow rate of fluid from any one of the at least two bioreactor stations to any one of the at least two harvest subsystems.

21. The facility may further include a logistics corridor, a staff area, a packaging area, a shipping area, and / or a receiving area. The building according to claim 15.

22. 16. The building of claim 15, configured to meet Leadership in Energy and Environmental Design (LEED) certification standards.

23. a plurality of motion, temperature, and / or humidity sensors distributed on, in, and / or around equipment within the building; and a monitoring system configured to track outputs of the plurality of sensors and to generate records and / or alarms in response to outputs that deviate from predefined ranges.

24. 1. A process for growing a biologic comprising: Growing the cell mass; (i) harvesting a cell or (ii) a material comprising a component of said cell; passing a first portion of the harvested material to a first purification subsystem; passing a second portion of the harvested material to a second purification subsystem; Producing a first product via the first purification subsystem; producing a second product via the second purification subsystem; The process includes:

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