Systems, devices, and methods for cell culture

Abstract

The embodiments described herein generally relate to assemblies, systems, and methods for cell culture and the production of biopharmaceuticals. A multi-purpose assembly is disclosed herein that facilitates a combination of a perfusion process and a collection process. The multi-purpose assembly and system are the same and simplify and streamline the biomanufacturing process, including by improving at least automation capabilities, reducing the physical footprint, reducing the risk of contamination, and reducing the rate of use of consumables. Methods of using the foregoing assemblies and systems are also disclosed herein.

Description

Technical Field

[0001] Cross-reference This application is a PCT application claiming the benefit of U.S. Application No. 63 / 357,593, filed Jun. 30, 2022, entitled "SYSTEMS, APPARATUS, AND METHODS FOR CELL CULTURE", which is hereby incorporated by reference in its entirety for all purposes.

[0002] The present disclosure generally relates to devices, systems, and methods for the production of biopharmaceuticals. More particularly, the present disclosure relates to the improvement of the utilization of cell retention devices in devices, systems, and methods for biomanufacturing.

Background Art

[0003] Cell culture, e.g., the culture of mammalian, bacterial, or fungal cells, can be performed to collect live cells for therapeutic purposes and / or to collect biological molecules such as proteins or chemicals (e.g., biopharmaceuticals) produced by the cells. During the process of biomanufacturing, including cell culture, filtration is performed to separate, clarify, modify, and / or concentrate fluid solutions, mixtures, or suspensions. In the biotechnology and pharmaceutical industries, filtration is essential for the production, processing, and testing of new drugs, diagnostics, and other biopharmaceuticals. For example, in the process of manufacturing biopharmaceuticals using animal or microbial cell cultures, filtration is performed for the clarification, selective removal, and / or concentration of specific components from the culture medium and / or to modify the medium prior to further processing. Filtration can also be used to increase productivity by maintaining the culture at a high cell concentration perfusion.

[0004] One use of the filtration unit can be a cell retention device (CRD). Note-worthy CRDs include tangential flow filtration (also called cross-flow filtration or TFF) systems. Such systems are widely used for the separation of fine particles suspended in a liquid phase and have important biomanufacturing applications. In contrast to dead-end filtration systems where a single fluid feed stream passes through a filter, tangential flow systems are characterized by a fluid feed stream that flows across the surface of the filter and is separated into two components, a permeate component that passes through the filter and a retentate component that does not. Compared to dead-end systems, TFF systems are less prone to fouling. Fouling of TFF systems can be further reduced by alternating the direction of the fluid feed stream across the filter elements as done in alternating tangential flow (ATF) technology, by backwashing the permeate passing through the filter, and / or by periodically cleaning the filter.

[0005] Advances in filtration have been made (see, for example, Tim Sewerin et al., “Advances and Applications of Hollow Fiber Nanofiltration Membranes: A Review. Membranes”, 2021, 11, 890). One such advance includes hollow fiber (HF) filters (HFFs) that can operate in tangential flow mode, an example of which is the tangential flow depth filtration (TM) (TFDF(TM)) filter. Such TFDF(TM) filters combine the advantages of tangential flow and depth filtration. However, certain HF filters are considered expensive and / or relatively bulky, and cell cultures are prone to contamination and loss of associated products during transfer between bioreactors for the cell growth and collection stages.

[0006] Therefore, it is desirable to have assemblies, systems, and methods that can improve these processes. SUMMARY OF THE INVENTION

[0007] Assemblies, systems, and methods suitable for the production of biologic agents are described herein. The bioreactor system disclosed herein includes: a) a bioreactor including an input port and an output port; b) a supply flow conduit operably connected to the output port and the inlet of a hollow fiber (HF) filter, the supply flow conduit being operable to carry fluid from the bioreactor to the HF filter; c) a hold-up liquid flow conduit operably connected to the input port of the bioreactor and the outlet of the HF filter, the hold-up liquid flow conduit being operable to carry fluid from the HF filter to the bioreactor; and d) a multipurpose assembly operably connected to the HF filter, the multipurpose assembly including a first flow path and a second flow path, the first flow path including a first pump operable to draw fluid from the HF filter and a flow meter operable to measure the flow rate of the fluid in the first flow path, the second flow path including a second pump operable to draw fluid from the HF filter, the first pump and the second pump being configured to have different pump capacities, a bioreactor system.

[0008] In some embodiments, the HF filter is operable with respect to tangential flow. In some embodiments, the HF filter is a tangential flow depth filtration (TFDF) filter. In some embodiments, the first and second pumps are configured to have different accuracy evaluations. In some embodiments, the system further includes one or more clarification filters in one or more of the first and / or second flow paths. In some embodiments, the first and second pumps are selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetic drive pumps, positive displacement pumps, membrane pumps, pressure pumps, Quantex (e.g., positive displacement rotary) pumps, gear pumps, diaphragm pumps, syringe pumps, and piston pumps. In some embodiments, the first pump and the second pump are peristaltic pumps. In some embodiments, the supply flow conduit comprises a supply flow pump selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetic drive pumps, positive displacement pumps, membrane pumps, pressure pumps, Quantex trademark pumps, gear pumps, diaphragm pumps, syringe pumps, and piston pumps, and is operable to carry unfiltered fluid from the bioreactor to the HF filter. In some embodiments, the hold liquid flow conduit is operable to carry cell culture fluid from the HF filter to the bioreactor.

[0009] In some embodiments, the first flow path including a first pump operable to draw fluid from the HF filter draws perfusion permeate flow fluid from the HF filter. In some embodiments, the second flow path including a second pump operable to draw fluid from the HF filter draws collection permeate flow fluid from the HF filter. In some embodiments, the first flow path and the second flow path have different inner diameters. In some embodiments, the first flow path and the second flow path have the same inner diameter. In some embodiments, the second flow path can direct fluid at a higher flow rate than the first flow path.

[0010] In some embodiments, the first pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 liters per minute (LPM) to 5 LPM, about 0.01 LPM to about 10 LPM, and / or about 0.01 LPM to about 15 LPM. In some embodiments, the first flow path and pump are operable to draw a feed flow rate of about 0.8 to about 2.2 liters per fiber per minute (L / fiber / min) from the HF filter. In some embodiments, the first pump is about 75×10 6 cells / mL or greater, about 50×10 6 cells / mL or greater, and / or about 25×10 6 operable to draw fluid from the HF filter while maintaining a viable cell density (VCD) of cells / mL or greater. In some embodiments, the first pump is operable to draw fluid from the HF filter at a rate of about 0.1 to about 5 bioreactor vessel volumes per day (VVD), about 0.5 to about 4.3 bioreactor VVD, about 1 to about 4.3 bioreactor VVD, and / or a maximum of about 5 bioreactor VVD. In some embodiments, the first pump is operable to draw fluid from the HF filter at a filter flow rate of about 50 to about 800 liters per square meter per hour (LMH), and / or about 100 to about 600 LMH.

[0011] In some embodiments, a flow meter operable to measure the flow rate of the fluid in the first flow path continuously monitors the VVD rate and communicates with the first pump to adjust the first pump speed to maintain the desired VVD rate. In some embodiments, a flow meter operable to measure the flow rate of the fluid in the first flow path can accurately monitor a flow rate of about 0 to about 8 LPM, or about 0.5 LPM to about 6 LPM. In some embodiments, the first pump is about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 , about 10,000 liters per square meter (L / m 2 ) to about 50,000 L / m 2 , and / or about 10,000 liters per square meter (L / m 2) to approximately 70,000 L / m 2 It is operable to draw fluid from the HF filter at a throughput equal to 2 . In some embodiments, the first pump is operable to draw fluid from the HF filter while maintaining a shear rate (s-1) of less than about 5,000 s-1, less than about 3,500 s-1, and / or less than about 2,500 s-1.

[0012] In some embodiments, the bioreactor has a volume of about 15 liters or less, about 50 liters or less, about 100 liters or less, or about 500 liters or less. In some embodiments, the bioreactor has a volume of about 500 liters, about 1,000 liters, about 1,500 liters, and / or about 3,500 liters or more. In some embodiments, the bioreactor has a volume equal to about 2,000 liters to about 7,000 liters. In some embodiments, the bioreactor has at least two input ports and at least two output ports and at least two of b), c), and d).

[0013] In some embodiments, the bioreactor system can operate in a perfusion process. In some embodiments, the bioreactor system can operate in a collection process. In some embodiments, the bioreactor system can operate in a perfusion process, a process between perfusion and collection, and a collection process. In some embodiments, the bioreactor system can operate in a continuous collection process. In some embodiments, the bioreactor system further comprises a human machine interface (HMI) control unit. In some embodiments, the HMI control unit can be programmed to display perfusion process control, control of the process between perfusion and collection, or collection process control.

[0014] In some embodiments, the bioreactor system further comprises one or more sensors. In some embodiments, the bioreactor system further includes one or more of a feed stream or a holding liquid stream fluid pump. In some embodiments, the bioreactor system further includes at least one component capable of facilitating at least one process intensification parameter. In some embodiments, the process intensification parameter is one or more of an increase in cell number, an increase in cell density, a supply of a rich cell culture growth medium, a rapid expansion of cell number, or an increase in the production of a biologic. In some embodiments, the bioreactor system is further capable of backflushing the HF filter with permeate fluid.

[0015] In some embodiments, the bioreactor system further comprises a second flow meter operable to measure the flow rate of the fluid in the second flow path and accurately monitor a flow rate of from about 0 to about 10 LPM, or from about 0.5 LPM to about 8 LPM. In some embodiments, the second pump is operable to draw fluid from the HF filter at a throughput equal to from about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 , from about 12,000 liters per square meter (L / m 2 ) to about 60,000 L / m 2 , and / or from about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 . In some embodiments, the second pump is operable to draw fluid from the HF filter at a filter flux rate of from about 150 to about 900 liters per square meter per hour (LMH), and / or from about 200 to about 700 LMH.

[0016] In some embodiments, the second flow path and the pump are operable to draw a supply flow rate of from about 1 to about 3 liters per fiber per minute (L / fiber / min) from the HF filter. In some embodiments, the second pump is operable to draw fluid from the HF filter at a flow rate of from about 0.01 LPM to about 8 LPM, from about 0.01 LPM to about 13 LPM, and / or from about 0.01 LPM to about 18 LPM.

[0017] In some embodiments, the first flow path comprises a connection system having 3 / 8 inch ID tubing and / or 1 / 8 inch ID tubing. In some embodiments, the second flow path comprises a connection system having 1 / 2 inch ID tubing and / or 1 / 8 inch ID tubing. In some embodiments, the first flow path has an accuracy requirement of about 1%. In some embodiments, the second flow path has an accuracy requirement of about 3%.

[0018] In some embodiments, the HF filter is composed of, and / or includes, polypropylene and / or polyethylene terephthalate. In some embodiments, the HF filter includes an isotropic pore structure. In some embodiments, the HF filter has an average pore size of from about 0.65 μm to about 8 μm. In some embodiments, the HF filter has an average pore size of from about 2 μm to about 5 μm. In some embodiments, the HF filter has an average pore size of about 2 μm.

[0019] In some embodiments, the bioreactor system is operable for use with shear-sensitive cells. In some embodiments, the bioreactor system is operable for use with animal cells. In some embodiments, the bioreactor system is operable for use with mammalian cells. In some embodiments, the bioreactor system is operable for use with Chinese hamster ovary (CHO) cells. In some embodiments, the bioreactor system is operable for use with human embryonic kidney 293 (HEK293) cells. In some embodiments, the bioreactor system is operable for use with cells for the production of biopharmaceuticals. In some embodiments, the biopharmaceuticals include antibodies, peptides, and / or viruses.

[0020] Also disclosed herein is a multipurpose filter assembly comprising: a) a hollow fiber (HF) filter; and b) a multipurpose assembly operably connected to the HF filter and including a first flow path and a second flow path, the first flow path including a first pump operable to draw fluid from the HF filter and a flow meter operable to measure the flow rate of the fluid in the first flow path, the second flow path including a second pump operable to draw fluid from the HF filter, the first pump and the second pump being capable of having different pump capacities and accuracy ratings. In some embodiments, the HF filter is operable for tangential flow.

[0021] In some embodiments, the HF filter is a tangential flow depth filtration (registered trademark) (TFDF (registered trademark)) filter. In some embodiments, the multi-purpose filter assembly further comprises one or more clarification filters in the first and / or second flow paths. In some embodiments, the first and second pumps are selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetic drive pumps, positive displacement pumps, membrane pumps, pressure pumps, Quantex (trademark) pumps, gear pumps, diaphragm pumps, syringe pumps, and piston pumps. In some embodiments, the first pump and the second pump are peristaltic pumps. In some embodiments, the first flow path and pump are operable to draw fluid from the HF filter at a throughput of about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 or more. In some embodiments, the first flow path and pump are operable to draw fluid from the HF filter at a throughput of about 10,000 liters per square meter (L / m 2 ) to about 50,000 L / m 2 or more. In some embodiments, the first flow path and pump are operable to draw fluid from the HF filter at a throughput of about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2It is operable to draw fluid from the HF filter at the throughput described above. In some embodiments, the first flow path and the pump are operable to draw a feed flow rate of from about 0.8 to about 2.2 liters per fiber per minute (L / fiber / min) from the HF filter. In some embodiments, the first flow path and the pump are operable to draw a feed flow rate of from about 0.8 to about 2.0 L / fiber / min and / or from about 1 to about 1.8 L / fiber / min from the HF filter. In some embodiments, the first flow path and the pump are operable to draw fluid from the HF filter at a filter flux of from about 50 to about 800 LMH and / or from about 100 to about 600 LMH. In some embodiments, the first flow path and the pump are operable to maintain a packed cell volume (PCV) in the blood of from about 2 to about 40%, from about 8 to about 40%, and / or from about 12 to about 35%. In some embodiments, the first flow path and the pump are operable to draw fluid from the HF filter while maintaining a viable cell density (VCD) of greater than about 25×10 6 cells / mL, about 50×10 6 cells / mL, about 75×10 6 cells / mL, about 100×10 6 cells / mL, and / or greater than about 25×10 7 cells / mL. In some embodiments, the first flow path and the pump are operable to draw fluid from the HF filter at a shear rate of less than about 5,000 s-1, about 3,500 s-1, and / or about 2,500 s-1. In some embodiments, the first flow path and the pump are operable to draw fluid from the HF filter at a flow rate of from about 0.01 LPM to about 5 LPM, from about 0.01 LPM to about 10 LPM, and / or from about 0.01 LPM to about 15 LPM.

[0022] In some embodiments, a flow meter operable to measure the flow rate of the fluid in the first flow path continuously monitors the LPM rate and communicates with the first pump to adjust the first pump speed to maintain the desired LPM rate. In some embodiments, a flow meter operable to measure the flow rate of the fluid in the first flow path can accurately monitor flow rates of from about 0 to about 8 LPM and / or from about 0.01 LPM to about 6 LPM. In some embodiments, the first flow path has an accuracy requirement of about 1%. In some embodiments, the second flow path has an accuracy requirement of about 3%.

[0023] In some embodiments, the first flow path and the second flow path have different inner diameters. In some embodiments, the first flow path and the second flow path have the same inner diameter. In some embodiments, the second flow path and pump are operable to draw fluid from the HF filter at a higher flow rate than the first flow path and the first pump. In some embodiments, the second flow path and pump are operable to draw fluid from the HF filter as a continuous collection process. In some embodiments, the HF filter is not replaced when the multi-purpose assembly directs fluid through one flow path and pump and then through the other flow path and pump. In some embodiments, the HF filter is not replaced when the multi-purpose assembly directs fluid through the first flow path and pump first and then through the second flow path and pump. In some embodiments, the first and second flow paths of the multi-purpose assembly are connected by a T-connector, a Y-connector, or a valve.

[0024] In some embodiments, the multi-purpose filter assembly further comprises a second flow meter operable to measure the flow rate of the fluid in the second flow path and accurately monitor flow rates of from about 0 to about 10 LPM or from about 0.5 LPM to about 8 LPM. In some embodiments, the second pump is from about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 and from about 12,000 L / m 2 to about 60,000 L / m 2and / or about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 and is operable to draw fluid from the HF filter at a throughput equal thereto. In some embodiments, the second pump is operable to draw fluid from the HF filter at a filter flow rate of about 150 to about 900 liters per square meter per hour (LMH), and / or about 200 to about 700 LMH. In some embodiments, the second flow path and pump are operable to draw a feed flow rate of about 1 to about 3 liters per fiber per minute (L / fiber / min) from the HF filter. In some embodiments, the second pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 8 LPM, about 0.01 LPM to about 13 LPM, and / or about 0.01 LPM to about 18 LPM.

[0025] In some embodiments, the HF filter is composed of and / or includes polypropylene and / or polyethylene terephthalate. In some embodiments, the HF filter includes an isotropic pore structure. In some embodiments, the HF filter has an average pore size of about 0.65 μm to about 8 μm. In some embodiments, the HF filter has an average pore size of about 2 μm to about 5 μm. In some embodiments, the HF filter has an average pore size of about 2 μm.

[0026] In some embodiments, the multi-purpose filter assembly is operable for use with shear-sensitive cells, animal cells, mammalian cells, CHO cells, and / or HEK293 cells. In some embodiments, the multi-purpose filter assembly is operable for use with cells for the production of biopharmaceuticals. In some embodiments, the biopharmaceuticals include antibodies, peptides, and / or viruses.

[0027] Also provided herein is a method for producing a biologic from cells, comprising: a) growing cells in a cell culture fluid within a bioreactor; b) during cell growth, perfusing the cell culture fluid through a hollow fiber (HF) filter to remove spent cell culture medium while retaining the grown cells, and adding an appropriate replacement volume of cell culture medium to the bioreactor to maintain a desired cell culture fluid level within the bioreactor, wherein the spent cell culture medium is perfused through the HF filter at a first flow rate for entry into a first flow path; c) after cell growth, collecting the cell culture fluid through the HF filter to obtain the biologic, and adding an appropriate replacement volume of cell culture fluid to the bioreactor to maintain a desired cell culture fluid level within the bioreactor during different stages of collection, wherein the cell culture medium is collected through the HF filter at a second flow rate for entry into a second flow path, and the second flow rate is greater than the first flow rate. In some embodiments, the HF filter is operable with respect to tangential flow. In some embodiments, the HF filter is a tangential flow depth filtration (registered trademark) (TFDF (registered trademark)) filter. In some embodiments, the replacement volume of cell culture medium in b) and / or c) is the same amount of spent cell culture medium removed. In some embodiments, the replacement volume of cell culture medium in b) and / or c) is greater than the amount of spent cell culture medium removed. In some embodiments, the replacement volume of cell culture medium in b) and / or c) is less than the amount of spent cell culture medium removed. In some embodiments, the replacement volume of cell culture medium is fresh cell culture medium. In some embodiments, the first flow rate is about 5 LPM or less, and the second flow rate is greater than about 5 LPM. In some embodiments, the HF filter is composed of and / or includes polypropylene and / or polyethylene terephthalate. In some embodiments, the HF filter has an isotropic pore structure. In some embodiments, the HF filter has an average pore size of about 0.65 μm to about 8 μm. In some embodiments, the HF filter has an average pore size of about 2 μm to about 5 μm.In some embodiments, the HF filter has an average pore size of about 2 μm. In some embodiments, the cells are shear-sensitive cells. In some embodiments, the cells are animal cells, mammalian cells, CHO cells, and / or HEK293 cells. In some embodiments, the biologic agent includes an antibody, a peptide, and / or a virus.

[0028] Also disclosed herein is a method for producing a biologic agent from cells, including the use of the bioreactor system and / or the multi-purpose filter assembly disclosed herein.

[0029] Also disclosed herein is the use of the multi-purpose filter assembly disclosed herein in a bioreactor system. In some embodiments, the bioreactor system is the bioreactor system of the present disclosure or the present invention.

[0030] Also disclosed herein is the use of the bioreactor system disclosed herein for culturing cells. In some embodiments, the cells produce a biologic agent, and the use may further include collecting the cell culture through an HF filter to obtain the biologic agent.

[0031] Also disclosed herein is a method for producing a biologic agent from cells, including the use of the bioreactor system or the multi-purpose filter assembly disclosed herein. Further disclosed herein is a method of using the system, assembly, or method disclosed herein for improving productivity, performance, efficiency, reducing the contamination rate, reducing capital investment, reducing the physical footprint, reducing the consumption of consumables, and / or reducing the operating cost of a facility, including the production process of the biologic agent.

[0032] Certain embodiments of the present invention are characterized by the following aspects.

[0033] Aspect 1 is a bioreactor system, comprising: a) a bioreactor including an input port and an output port; b) a supply flow conduit operably connected to the output port and the inlet of a hollow fiber (HF) filter, the supply flow conduit being operable to carry fluid from the bioreactor to the HF filter; c) a holding liquid flow conduit operably connected to the input port of the bioreactor and the outlet of the HF filter, the holding liquid flow conduit being operable to carry fluid from the HF filter to the bioreactor; and d) a multi-purpose assembly operably connected to the HF filter, the multi-purpose assembly comprising a first flow path and a second flow path, the first flow path including a first pump operable to draw fluid from the HF filter and a flow meter operable to measure the flow rate of the fluid in the first flow path, the second flow path including a second pump operable to draw fluid from the HF filter, the first pump and the second pump being configured to have different pump capacities.

[0034] Aspect 2 is the bioreactor system according to Aspect 1, wherein the HF filter is operable for tangential flow.

[0035] Aspect 3 is the bioreactor system according to Aspect 1 or 2, wherein the HF filter is a tangential flow depth filtration (registered trademark) (TFDF (registered trademark)) filter.

[0036] Aspect 4 is the bioreactor system according to any one of Aspects 1 to 3, wherein the first and second pumps are configured to have different accuracy evaluations.

[0037] Aspect 5 is the bioreactor system according to any one of Aspects 1 to 4, further comprising one or more clarification filters in one or both of the first and / or second flow paths.

[0038] Aspect 6 is the bioreactor system according to any one of aspects 1 to 5, wherein the first and second pumps are selected from the group consisting of a peristaltic pump, a centrifugal pump, a magnetic drive pump, a positive displacement pump, a membrane pump, a pressure pump, a Quantex (trademark) pump, a gear pump, a diaphragm pump, a syringe pump, and a piston pump.

[0039] Aspect 7 is the bioreactor system according to any one of aspects 1 to 6, wherein the first pump and the second pump are peristaltic pumps.

[0040] Aspect 8 is the bioreactor system according to any one of aspects 1 to 7, wherein the supply flow conduit is provided with a supply flow pump selected from the group consisting of a peristaltic pump, a centrifugal pump, a magnetic drive pump, a positive displacement pump, a membrane pump, a pressure pump, a Quantex (trademark) pump, a gear pump, a diaphragm pump, a syringe pump, and a piston pump, and is operable to carry unfiltered fluid from the bioreactor to the HF filter.

[0041] Aspect 9 is the bioreactor system according to any one of aspects 1 to 8, wherein the holding liquid flow conduit is operable to carry the cell culture solution from the HF filter to the bioreactor.

[0042] Aspect 10 is the bioreactor system according to any one of aspects 1 to 9, wherein the first flow path including the first pump operable to draw fluid from the HF filter draws the perfusion permeate fluid from the HF filter.

[0043] Aspect 11 is the bioreactor system according to any one of aspects 1 to 10, wherein the second flow path including the second pump operable to draw fluid from the HF filter draws the collected permeate fluid from the HF filter.

[0044] Aspect 12 is the bioreactor system according to any one of aspects 1 to 11, wherein the first flow path and the second flow path have the same inner diameter or different inner diameters.

[0045] Aspect 13 is the bioreactor system according to any one of Aspects 1 to 12, wherein the second flow path can induce a fluid at a higher flow rate than the first flow path.

[0046] Aspect 14 is the bioreactor system according to any one of Aspects 1 to 13, wherein the first pump is operable to draw a fluid from the HF filter at a flow rate of about 0.01 liters per minute (LPM) to 5 LPM.

[0047] Aspect 15 is the bioreactor system according to any one of Aspects 1 to 13, wherein the first pump is operable to draw a fluid from the HF filter at a flow rate of about 0.01 LPM to about 10 LPM.

[0048] Aspect 16 is the bioreactor system according to any one of Aspects 1 to 13, wherein the first pump is operable to draw a fluid from the HF filter at a flow rate of about 0.01 LPM to about 15 LPM.

[0049] Aspect 17 is the bioreactor system according to any one of Aspects 1 to 16, wherein the first flow path and the pump are operable to draw a supply flow rate of about 0.8 to about 2.2 liters per fiber per minute (L / fiber / min) from the HF filter.

[0050] Aspect 18 is such that the first pump is operable to draw a fluid from the HF filter while maintaining a viable cell density (VCD) of more than about 75×10 6 cells / mL, and is the bioreactor system according to any one of Aspects 1 to 17.

[0051] Aspect 19 is such that the first pump is operable to draw a fluid from the HF filter while maintaining a viable cell density (VCD) of more than about 50×10 6 cells / mL, and is the bioreactor system according to any one of Aspects 1 to 17.

[0052] Aspect 20 is the bioreactor system according to any one of aspects 1 to 17, wherein the first pump is operable to draw fluid from the HF filter while maintaining a VCD of more than about 25×10 6 cells / mL.

[0053] Aspect 21 is the bioreactor system according to any one of aspects 1 to 20, wherein the first pump is operable to draw fluid from the HF filter at a rate of about 0.1 to about 5 bioreactor vessel volumes per day (VVD).

[0054] Aspect 22 is the bioreactor system according to any one of aspects 1 to 21, wherein the first pump is operable to draw fluid from the HF filter at a rate of about 0.5 to about 4.3 bioreactor vessel volumes per day (VVD).

[0055] Aspect 23 is the bioreactor system according to any one of aspects 1 to 22, wherein the first pump is operable to draw fluid from the HF filter at a filter flow rate of about 50 to about 800 liters per square meter per hour (LMH).

[0056] Aspect 24 is the bioreactor system according to any one of aspects 1 to 23, wherein the first pump is operable to draw fluid from the HF filter at a filter flow rate of about 100 to about 600 LMH.

[0057] Aspect 25 is the bioreactor system according to any one of aspects 1 to 24, wherein a flow meter operable to measure the flow rate of the fluid in the first flow path continuously monitors the VVD rate and communicates with the first pump to adjust the first pump rate to maintain the desired VVD rate.

[0058] Aspect 26 is the bioreactor system according to any one of aspects 1 to 25, wherein a flow meter operable to measure the flow rate of the fluid in the first flow path can accurately monitor a flow rate of about 0 to about 8 LPM, or about 0.5 LPM to about 6 LPM.

[0059] Aspect 27 is the bioreactor system according to any one of aspects 1 to 26, wherein the first pump is operable to draw fluid from the HF filter at a throughput equal to about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 .

[0060] Aspect 28 is the bioreactor system according to any one of aspects 1 to 26, wherein the first pump is operable to draw fluid from the HF filter at a throughput equal to about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 .

[0061] Aspect 29 is the bioreactor system according to any one of aspects 1 to 28, wherein the first pump is operable to draw fluid from the HF filter while maintaining a shear rate (s-1) of less than about 5,000 s-1

[0062] Aspect 30 is the bioreactor system according to any one of aspects 1 to 29, wherein the first pump is operable to draw fluid from the HF filter while maintaining a shear rate (s-1) of less than about 3,500 s-1

[0063] Aspect 31 is the bioreactor system according to any one of aspects 1 to 30, wherein the first pump is operable to draw fluid from the HF filter while maintaining a shear rate (s-1) of less than about 2,500 s-1

[0064] Aspect 32 is the bioreactor system according to any one of aspects 1 to 31, wherein the bioreactor has a volume of about 15 liters or less, about 50 liters or less, about 100 liters or less, or about 500 liters or less

[0065] Aspect 33 is the bioreactor system according to any one of Aspects 1 to 31, wherein the bioreactor has a volume of about 500 liters or more.

[0066] Aspect 34 is the bioreactor system according to any one of Aspects 1 to 31, wherein the bioreactor has a volume of about 1,000 liters or more.

[0067] Aspect 35 is the bioreactor system according to any one of Aspects 1 to 31, wherein the bioreactor has a volume of about 1,500 liters or more.

[0068] Aspect 36 is the bioreactor system according to any one of Aspects 1 to 31, wherein the bioreactor has a volume equal to about 2,000 liters to about 3,500 liters.

[0069] Aspect 37 is the bioreactor system according to any one of Aspects 1 to 31, wherein the bioreactor has a volume equal to about 2,000 liters to about 7,000 liters.

[0070] Aspect 38 is the bioreactor system according to any one of Aspects 1 to 37, wherein the bioreactor has at least two input ports and at least two output ports, and at least two of b), c), and d).

[0071] Aspect 39 is the bioreactor system according to any one of Aspects 1 to 38, wherein the bioreactor system can operate in a perfusion process.

[0072] Aspect 40 is the bioreactor system according to any one of Aspects 1 to 39, wherein the bioreactor system can operate in a collection process.

[0073] Aspect 41 is the bioreactor system according to any one of Aspects 1 to 40, wherein the bioreactor system can operate in a perfusion process, a process between perfusion and collection, and a collection process.

[0074] Aspect 41.1 is the bioreactor system according to any one of Aspects 1 to 41, wherein the bioreactor system can operate in two or more perfusion processes.

[0075] Aspect 41.2 is the bioreactor system according to any one of Aspects 1 to 41.1, wherein the bioreactor system can operate in two or more perfusion processes, a process between perfusion and collection, and a collection process.

[0076] Aspect 42 is the bioreactor system according to any one of Aspects 1 to 41.2, wherein the bioreactor system can operate in a continuous collection process.

[0077] Aspect 43 is the bioreactor system according to any one of Aspects 1 to 42, further comprising a human-machine interface (HMI) control unit.

[0078] Aspect 44 is the bioreactor system according to Aspect 43, wherein the HMI control unit is programmed to display perfusion process control, control of the process between perfusion and collection, or collection process control.

[0079] Aspect 45 is the bioreactor system according to any one of Aspects 1 to 44, further comprising one or more sensors.

[0080] Aspect 46 is the bioreactor system according to any one of Aspects 1 to 46, further comprising one or more of a supply flow or a holding liquid flow fluid pump.

[0081] Aspect 47 is the bioreactor system according to any one of aspects 1 to 46, further comprising at least one component capable of promoting at least one process intensification parameter.

[0082] Aspect 48 is the bioreactor system according to aspect 47, wherein the process intensification parameter is one or more of an increase in cell number, an increase in cell density, a supply of a rich cell culture growth medium, a rapid expansion of cell number, or an increase in the production of a biopharmaceutical.

[0083] Aspect 49 is the bioreactor system according to any one of aspects 1 to 48, further capable of backflushing the HF filter with the permeate fluid.

[0084] Aspect 50 is the bioreactor system according to any one of aspects 1 to 49, further comprising a second flow meter operable to measure the flow rate of the fluid in the second flow path and accurately monitor a flow rate of about 0 to about 10 LPM, or about 0.5 LPM to about 8 LPM.

[0085] Aspect 51 is the bioreactor system according to any one of aspects 1 to 50, wherein the second pump is operable to draw fluid from the HF filter at a throughput equal to about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 .

[0086] Aspect 52 is the bioreactor system according to any one of aspects 1 to 50, wherein the second pump is operable to draw fluid from the HF filter at a throughput equal to about 12,000 liters per square meter (L / m 2 ) to about 60,000 L / m 2 , and / or at a throughput equal to about 12,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 .

[0087] Aspect 53 is the bioreactor system according to any one of aspects 1 to 52, wherein the second pump is operable to draw fluid from the HF filter at a filter flow rate of about 150 to about 900 liters per square meter per hour (LMH).

[0088] Aspect 54 is the bioreactor system according to any one of aspects 1 to 53, wherein the second pump is operable to draw fluid from the HF filter at a filter flow rate of about 200 to about 700 LMH.

[0089] Aspect 55 is the bioreactor system according to any one of aspects 1 to 54, wherein the second flow path and the pump are operable to draw a supply flow rate of about 1 to about 3 liters per fiber per minute (L / fiber / min) from the HF filter.

[0090] Aspect 56 is the bioreactor system according to any one of aspects 1 to 55, wherein the second pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 8 LPM.

[0091] Aspect 57 is the bioreactor system according to any one of aspects 1 to 55, wherein the second pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 13 LPM.

[0092] Aspect 58 is the bioreactor system according to any one of aspects 1 to 55, wherein the second pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 18 LPM.

[0093] Aspect 59 is the bioreactor system according to any one of aspects 1 to 58, wherein the first flow path comprises a connection system having a 3 / 8 inch ID tube.

[0094] Aspect 60 is the bioreactor system according to any one of aspects 1 to 58, wherein the first flow path comprises a connection system having a pipe with an ID of 1 / 8 inch.

[0095] Aspect 61 is the bioreactor system according to any one of aspects 1 to 60, wherein the second flow path comprises a connection system having a pipe with an ID of 1 / 2 inch.

[0096] Aspect 62 is the bioreactor system according to any one of aspects 1 to 60, wherein the second flow path comprises a connection system having a pipe with an ID of 1 / 8 inch.

[0097] Aspect 63 is the bioreactor system according to any one of aspects 1 to 62, wherein the first flow path has an accuracy requirement of about 1%.

[0098] Aspect 64 is the bioreactor system according to any one of aspects 1 to 63, wherein the second flow path has an accuracy requirement of about 3%.

[0099] Aspect 65 is the bioreactor system according to any one of aspects 1 to 64, wherein the HF filter is constructed of and / or includes polypropylene and / or polyethylene terephthalate.

[0100] Aspect 66 is the bioreactor system according to any one of aspects 1 to 65, wherein the HF filter includes an isotropic pore structure and / or has an average pore lumen diameter of about 0.65 μm to about 8 μm, or about 2 μm to about 5 μm.

[0101] Aspect 67 is the bioreactor system according to any one of aspects 1 to 66, which is operable for use with shear-sensitive cells.

[0102] Aspect 68 is the bioreactor system according to any one of aspects 1 to 67, which is operable for use with animal cells.

[0103] Aspect 69 is the bioreactor system according to any one of aspects 1 to 68, which is operable for use with mammalian cells.

[0104] Aspect 70 is the bioreactor system according to any one of aspects 1 to 69, which is operable for use with Chinese hamster ovary (CHO) cells.

[0105] Aspect 71 is the bioreactor system according to any one of aspects 1 to 69, which is operable for use with human embryonic kidney 293 (HEK293) cells.

[0106] Aspect 72 is the bioreactor system according to any one of aspects 1 to 71, which is operable for use with cells for the production of biopharmaceuticals.

[0107] Aspect 73 is the bioreactor system according to aspect 72, wherein the biopharmaceutical comprises an antibody, a peptide, and / or a virus.

[0108] Aspect 74 is a multipurpose filter assembly comprising: a) a hollow fiber (HF) filter, and b) a multipurpose assembly operably connected to the HF filter, the multipurpose assembly including a first flow path and a second flow path, the first flow path including a first pump operable to draw fluid from the HF filter and a flow meter operable to measure the flow rate of the fluid in the first flow path, the second flow path including a second pump operable to draw fluid from the HF filter, and the first pump and the second pump being capable of having different pump capacities and accuracy ratings.

[0109] Aspect 75 is the multipurpose filter assembly according to aspect 74, wherein the HF filter is operable for tangential flow.

[0110] Aspect 76 is the multipurpose filter assembly according to aspect 74 or 75, wherein the HF filter is a tangential flow depth filtration (registered trademark) (TFDF (registered trademark)) filter.

[0111] Aspect 77 is the multipurpose filter assembly according to any one of aspects 74 to 76, wherein the multipurpose filter assembly further includes one or more clarification filters in the first and / or second flow paths.

[0112] Aspect 78 is the multipurpose filter assembly according to any one of aspects 74 to 77, wherein the first and second pumps are selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetic drive pumps, positive displacement pumps, membrane pumps, pressure pumps, Quantex (trademark) pumps, gear pumps, diaphragm pumps, syringes, and piston pumps.

[0113] Aspect 79 is the multipurpose filter assembly according to any one of aspects 74 to 78, wherein the first pump and the second pump are peristaltic pumps.

[0114] Aspect 80 is the multipurpose filter assembly according to any one of aspects 74 to 79, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a throughput of about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 or more.

[0115] Aspect 81 is the multipurpose filter assembly according to any one of aspects 74 to 79, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a throughput of about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 or more.

[0116] Aspect 82 is the multi-purpose filter assembly according to any one of aspects 74 to 81, wherein the first flow path and the pump are operable to draw a supply flow rate of about 0.8 to about 2.2 liters per fiber per minute (L / fiber / min) from the HF filter.

[0117] Aspect 83 is the multi-purpose filter assembly according to any one of aspects 74 to 82, wherein the first flow path and the pump are operable to draw a supply flow rate of about 0.8 to about 2.0 L / fiber / min from the HF filter.

[0118] Aspect 84 is the multi-purpose filter assembly according to any one of aspects 74 to 83, wherein the first flow path and the pump are operable to draw a supply flow rate of about 1 to about 1.8 L / fiber / min from the HF filter.

[0119] Aspect 85 is the multi-purpose filter assembly according to any one of aspects 74 to 84, wherein the first flow path and the pump are operable to draw fluid from the HF filter with a filter flow of about 50 to about 800 LMH.

[0120] Aspect 86 is the multi-purpose filter assembly according to any one of aspects 74 to 85, wherein the first flow path and the pump are operable to draw fluid from the HF filter with a filter flow of about 100 to about 600 LMH.

[0121] Aspect 87 is the multi-purpose filter assembly according to any one of aspects 74 to 86, wherein the first flow path and the pump are operable to maintain a blood packed cell volume (PCV) of about 2 to about 40%.

[0122] Aspect 88 is the multi-purpose filter assembly according to any one of aspects 74 to 87, wherein the first flow path and the pump are operable to maintain a PCV of about 8 to about 40%.

[0123] Aspect 89 is the multi-purpose filter assembly according to any one of aspects 74 to 88, wherein the first flow path and the pump are operable to maintain a PCV of about 12% to about 35%.

[0124] Aspect 90 is the multi-purpose filter assembly according to any one of aspects 74 to 89, wherein the first flow path and the pump are operable to draw fluid from the HF filter while maintaining a VCD of more than about 25×10 6 cells / mL.

[0125] Aspect 91 is the multi-purpose filter assembly according to any one of aspects 74 to 90, wherein the first flow path and the pump are operable to draw fluid from the HF filter while maintaining a VCD of more than about 50×10 6 cells / mL.

[0126] Aspect 92 is the multi-purpose filter assembly according to any one of aspects 74 to 91, wherein the first flow path and the pump are operable to draw fluid from the HF filter while maintaining a VCD of more than about 75×10 6 cells / mL.

[0127] Aspect 93 is the multi-purpose filter assembly according to any one of aspects 74 to 92, wherein the first flow path and the pump are operable to draw fluid from the HF filter while maintaining a VCD of more than about 100×10 6 cells / mL.

[0128] Aspect 94 is the multi-purpose filter assembly according to any one of aspects 74 to 93, wherein the first flow path and the pump are operable to draw fluid from the HF filter while maintaining a VCD of more than about 25×10 7 cells / mL.

[0129] Aspect 95 is the multi-purpose filter assembly according to any one of aspects 74 to 94, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a shear rate of less than about 5,000 s-1.

[0130] Aspect 96 is the multi-purpose filter assembly according to any one of aspects 74 to 95, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a shear rate of less than about 3,500 s-1.

[0131] Aspect 97 is the multi-purpose filter assembly according to any one of aspects 74 to 96, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a shear rate of less than about 2,500 s-1.

[0132] Aspect 98 is the multi-purpose filter assembly according to any one of aspects 74 to 97, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 5 LPM.

[0133] Aspect 99 is the multi-purpose filter assembly according to any one of aspects 74 to 97, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 10 LPM.

[0134] Aspect 100 is the multi-purpose filter assembly according to any one of aspects 74 to 97, wherein the first flow path and the pump are operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 15 LPM.

[0135] Aspect 101 is the multi-purpose filter assembly according to any one of aspects 74 to 100, wherein a flow meter operable to measure the flow rate of the fluid in the first flow path continuously monitors the LPM rate and communicates with the first pump to adjust the first pump speed to maintain the desired LPM rate.

[0136] Aspect 102 is the multi-purpose filter assembly according to any one of aspects 74 to 101, wherein a flow meter operable to measure the flow rate of the fluid in the first flow path can accurately monitor a flow rate of about 0 to about 8 LPM.

[0137] Aspect 103 is the multi-purpose filter assembly according to any one of aspects 74 to 102, wherein a flow meter operable to measure the flow rate of the fluid in the first flow path can accurately monitor a flow rate of about 0.01 LPM to about 6 LPM.

[0138] Aspect 104 is the multi-purpose filter assembly according to any one of aspects 74 to 103, wherein the first flow path has an accuracy requirement of about 1%.

[0139] Aspect 105 is the multi-purpose filter assembly according to any one of aspects 74 to 104, wherein the second flow path has an accuracy requirement of about 3%.

[0140] Aspect 106 is the multi-purpose filter assembly according to any one of aspects 74 to 105, wherein the first flow path and the second flow path have the same inner diameter or different inner diameters.

[0141] Aspect 107 is the multi-purpose filter assembly according to any one of aspects 74 to 106, wherein the second flow path and the pump are operable to draw fluid from the HF filter at a higher flow rate than the first flow path and the first pump.

[0142] Aspect 108 is the multi-purpose filter assembly according to any one of aspects 74 to 107, wherein the second flow path and the pump are operable to draw fluid from the HF filter as a continuous collection process.

[0143] Aspect 109 is the multi-purpose filter assembly according to any one of aspects 74 to 108, wherein the HF filter is not replaced when the multi-purpose assembly guides the fluid through one flow path and pump and then through the other flow path and pump.

[0144] Aspect 110 is the multi-purpose filter assembly according to Aspect 109, where the HF filter is not replaced when the multi-purpose assembly guides the fluid to first pass through the first flow path and the pump, and then through the second flow path and the pump.

[0145] Aspect 111 is the multi-purpose filter assembly according to any one of Aspects 74 to 110, where the first flow path and the second flow path of the multi-purpose assembly are connected by a T-connector, a Y-connector, or a valve.

[0146] Aspect 112 is the multi-purpose filter assembly according to any one of Aspects 74 to 111, further comprising a second flow meter operable to measure the flow rate of the fluid in the second flow path and accurately monitor a flow rate of about 0 to about 10 LPM, or about 0.5 LPM to about 8 LPM.

[0147] Aspect 113 is the multi-purpose filter assembly according to any one of Aspects 74 to 112, where the second pump is operable to draw fluid from the HF filter at a throughput equal to about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 .

[0148] Aspect 114 is the multi-purpose filter assembly according to any one of Aspects 74 to 112, where the second pump is operable to draw fluid from the HF filter at a throughput equal to about 12,000 liters per square meter (L / m 2 ) to about 60,000 L / m 2 , and / or about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 .

[0149] Aspect 115 is the multi-purpose filter assembly according to any one of aspects 74 to 114, wherein the second pump is operable to draw fluid from the HF filter at a filter flow rate of about 150 to about 900 liters per square meter per hour (LMH).

[0150] Aspect 116 is the multi-purpose filter assembly according to any one of aspects 74 to 115, wherein the second pump is operable to draw fluid from the HF filter at a filter flow rate of about 200 to about 700 LMH.

[0151] Aspect 117 is the multi-purpose filter assembly according to any one of aspects 74 to 116, wherein the second flow path and pump are operable to draw a supply flow rate of about 1 to about 3 liters per fiber per minute (L / fiber / min) from the HF filter.

[0152] Aspect 118 is the multi-purpose filter assembly according to any one of aspects 74 to 117, wherein the second pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 8 LPM.

[0153] Aspect 119 is the multi-purpose filter assembly according to any one of aspects 74 to 117, wherein the second pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 13 LPM.

[0154] Aspect 120 is the multi-purpose filter assembly according to any one of aspects 74 to 117, wherein the second pump is operable to draw fluid from the HF filter at a flow rate of about 0.01 LPM to about 18 LPM.

[0155] Aspect 121 is the multi-purpose filter assembly according to any one of aspects 74 to 120, wherein the HF filter is constructed of and / or includes polypropylene and / or polyethylene terephthalate.

[0156] Aspect 122 is the multi-purpose filter assembly according to any one of Aspects 74 to 121, wherein the HF filter includes an isotropic pore structure and / or has an average pore lumen diameter of about 0.65 μm to about 8 μm, or about 2 μm to about 5 μm.

[0157] Aspect 123 is the multi-purpose filter assembly according to any one of Aspects 74 to 122, which is operable for use with shear-sensitive cells.

[0158] Aspect 124 is the multi-purpose filter assembly according to any one of Aspects 74 to 123, which is operable for use with animal cells.

[0159] Aspect 125 is the multi-purpose filter assembly according to any one of Aspects 74 to 124, which is operable for use with mammalian cells.

[0160] Aspect 126 is the multi-purpose filter assembly according to any one of Aspects 74 to 125, which is operable for use with Chinese hamster ovary (CHO) cells.

[0161] Aspect 127 is the multi-purpose filter assembly according to any one of Aspects 74 to 125, which is operable for use with human embryonic kidney 293 (HEK293) cells.

[0162] Aspect 128 is the multi-purpose filter assembly according to any one of Aspects 74 to 127, which is operable for use with cells for the production of biopharmaceuticals.

[0163] Aspect 129 is the multi-purpose filter assembly according to Aspect 128, wherein the biopharmaceutical includes an antibody, a peptide, and / or a virus.

[0164] Aspect 130 is a method for producing a biologic from cells, comprising: a) growing cells in a cell culture fluid in a bioreactor; b) during cell growth, perfusing the cell culture fluid through a hollow fiber (HF) filter to remove spent cell culture medium while retaining the growing cells, and adding an appropriate replacement volume of cell culture medium to the bioreactor to maintain a desired cell culture fluid level in the bioreactor, wherein the spent cell culture medium is perfused through the HF filter at a first flow rate for entry into a first flow path; and c) subsequent to cell growth, collecting the cell culture fluid through the HF filter to obtain the biologic, and adding an appropriate replacement volume of cell culture medium to the bioreactor to maintain a desired cell culture fluid level in the bioreactor during different stages of collection, wherein the cell culture medium is collected through the HF filter at a second flow rate for entry into a second flow path, and the second flow rate is greater than the first flow rate.

[0165] Aspect 131 is the method according to aspect 130, wherein the HF filter is operable with respect to tangential flow.

[0166] Aspect 132 is the method according to any one of aspects 130 to 131, wherein the HF filter is a tangential flow depth filtration (TFDF) filter.

[0167] Aspect 133 is the method according to any one of aspects 130 to 132, wherein the replacement volume of cell culture medium in b) and / or c) is approximately the same amount of spent cell culture medium being removed.

[0168] Aspect 134 is the method according to any one of aspects 130 to 132, wherein the replacement volume of cell culture medium in b) and / or c) is greater than the amount of spent cell culture medium being removed.

[0169] Aspect 135 is the method according to any one of aspects 130 to 132, wherein the replacement volume of cell culture medium in b) and / or c) is less than the amount of spent cell culture medium being removed.

[0170] Aspect 136 is the method according to any one of Aspects 130 to 135, wherein the replacement amount of the cell culture medium is fresh cell culture medium.

[0171] Aspect 136.1 is the method according to any one of Aspects 130 to 136, wherein the perfusion process includes a plurality of perfusion stages.

[0172] Aspect 136.2 is the method according to any one of Aspects 130 to 136.1, wherein the perfusion process includes a plurality of HF filters in the perfusion stage, and is used in the plurality of perfusion stages, the process between perfusion and collection, and the collection process.

[0173] Aspect 137 is the method according to any one of Aspects 130 to 136.2, wherein the first flow rate is about 5 LPM or less and the second flow rate is greater than about 5 LPM.

[0174] Aspect 138 is the method according to any one of Aspects 130 to 137, wherein the HF filter is composed of and / or includes polypropylene and / or polyethylene terephthalate.

[0175] Aspect 139 is the method according to any one of Aspects 130 to 138, wherein the HF filter includes an isotropic pore structure and / or an average pore lumen diameter of about 0.65 μm to about 8 μm, or about 2 μm to about 5 μm.

[0176] Aspect 140 is the method according to any one of Aspects 130 to 139, wherein the cells are shear-sensitive cells.

[0177] Aspect 141 is the method according to any one of Aspects 130 to 140, wherein the cells are animal cells.

[0178] Aspect 142 is the method according to any one of Aspects 130 to 141, wherein the cells are mammalian cells.

[0179] Aspect 143 is the method according to any one of Aspects 130 to 142, wherein the cell is a CHO cell.

[0180] Aspect 144 is the method according to any one of Aspects 130 to 142, wherein the cell is a HEK293 cell.

[0181] Aspect 145 is the method according to any one of Aspects 130 to 144, wherein the biopharmaceutical comprises an antibody, a peptide, and / or a virus.

[0182] Aspect 146 is a method for producing a biopharmaceutical from a cell, comprising using the bioreactor system or the multipurpose filter assembly according to any one of Aspects 1 to 129.

[0183] Aspect 147 is the use of the multipurpose filter assembly according to any one of Aspects 74 to 129 in a bioreactor system.

[0184] Aspect 148 is the use according to Aspect 147, wherein the bioreactor system comprises the features according to any one of Aspects 1 to 73.

[0185] Aspect 149 is the use of the bioreactor system according to any one of Aspects 1 to 74 for culturing cells.

[0186] Aspect 150 is the use according to Aspect 149, wherein the cell produces a biopharmaceutical, and the use further comprises collecting the cell culture through an HF filter to obtain the biopharmaceutical.

[0187] Aspect 151 is the system, assembly, method, or use according to any one of Aspects 1 to 150, wherein the system, assembly, method, or use results in one or more of an improvement in productivity, an improvement in performance, an improvement in efficiency, a reduction in the contamination rate, a reduction in capital investment, a reduction in the physical footprint, a reduction in the consumption of consumables, or a reduction in the operating cost of the facility, the system, assembly, method, or use including a biopharmaceutical production process.

Brief Description of the Drawings

[0188] This disclosure is described in conjunction with the following accompanying drawings.

[0189]

Figure 1

[0190]

Figure 2

[0191]

Figure 3

Modes for Carrying Out the Invention

[0192] I. Overview The embodiments described herein recognize that biomanufacturing is an essential component of biopharmaceutical production and thus modern medicine.

[0193] Cell retention devices (CRDs) play an important role in establishing perfusion processes. Generally, during a perfusion process, a CRD enables retention of cells in a bioreactor while fresh medium is continuously supplied to the bioreactor and permeate (e.g., spent media) is continuously withdrawn at a predetermined rate. Currently commercially available cell retention devices (e.g., ATF and TFF) cannot operate successfully at high perfusion rates (e.g., 2 vessel volumes per day (VVD) or higher) for large-scale cell (e.g., CHO cell) culture working volumes (e.g., 3000 L or more). These cell retention devices have limitations regarding a flow of about 13 liters / m 2 / h, and a throughput of about 17,500 L / m 2 .

[0194] As described herein, in some embodiments, the inventors have shown that an HF filter (e.g., a TFDF® filter) can be used as a cell retention device for large-scale cell (e.g., CHO cell) culture working volumes (e.g., 3000 L or more) with high perfusion rates (e.g., 2 vessel volumes per day (VVD) or more), resulting in higher cell densities and titers. As shown herein, the inventors have created a perfusion-capable HF filter skid (e.g., a TFDF® filter skid) and successfully tested an HF filter (e.g., a TFDF® filter) having parameters of a flow of up to 600 liters / m 2 / h and a throughput of 70,000 L / m 2 . In some embodiments, the inventors have discovered that a system including TFDF® is suitable for operation at a bioreactor VVD of up to 5. Further, the inventors have found that in some embodiments, the same HF filter (e.g., a TFDF® filter) utilized during the perfusion process can also be used for collection processes such as, but not limited to, a continuous harvest (CH) process (also known as an extended harvest process, etc.). To the inventors' knowledge, this is the first demonstration that such a process can be carried out at such high pressures and / or volumes in process modes other than the collection mode. In some embodiments, the HF filter can be utilized for perfusion, batch, fed-batch, continuous, and / or semi-continuous harvest. In some methods including CH, while fresh cell culture medium (e.g., containing metabolites / buffers) is being perfused through the bioreactor prior to the end of a production run, the permeate from the HF filter is collected for downstream processing. However, after the production run, the addition of fresh cell culture medium (e.g., containing metabolites / buffers) is stopped and / or decreased while the permeate draw remains active, resulting in a decrease in the volume of the bioreactor. In some embodiments, all cell culture fluids and / or all permeates are processed in a downstream process (DSP).

[0195] In some embodiments, as described herein, while maintaining a low cell shear rate and / or cell stress level, compared to conventional systems, methods and / or apparatuses, a higher working volume (e.g., greater than 3,000 L), a higher VVD, an increased rate of medium exchange during perfusion, a higher flow, multi-stage use (e.g., one or more perfusion stages and one or more collection stages), and / or a higher throughput are achieved by using an HF filter, and thus, the culture of relatively sensitive cells (e.g., animal cells, e.g., cells lacking a cell wall) is facilitated without the cells passing through the filter to which they are associated.

[0196] In some embodiments, techniques such as multi-purpose assemblies and / or systems operable in high flow rates, large volumes, bioreactors of any size, and at any stage of the cell culture cycle are described herein. As described herein, in some embodiments, such capabilities (e.g., high flow rate, large volume, suitability for use in bioreactors of any size, and / or suitability for use at any stage of the cell culture cycle) are facilitated by features associated with hollow fiber filters and, in certain embodiments, hollow fiber filters operable in tangential flow mode.

[0197] In some embodiments, techniques such as multi-purpose assemblies that can be used to reduce the risk of contamination, reduce the rate of consumable depletion, reduce the physical footprint associated with biomanufacturing, enable continuous or semi-continuous bioprocesses for purification, and / or simplify biologic production processes are described herein.

[0198] In certain embodiments, techniques are provided herein for reducing the physical footprint associated with biomanufacturing (e.g., the manufacturing area required), e.g., reducing the required footprint by at least about 50%. In some embodiments, methods, systems, and apparatuses are provided herein that can reduce the physical footprint associated with biomanufacturing by at least about 66%. In some embodiments, the reduction in physical footprint is achieved by utilizing the systems and / or assemblies described herein to combine one or more biomanufacturing processes (e.g., perfusion processes and / or collection processes) in the same physical container and / or system, e.g., by using the same bioreactor for a perfusion process used in a collection process (e.g., batch, fed-batch, continuous, and / or semi-continuous collection).

[0199] In some embodiments, the techniques described herein provide additional automation features. For example, in some embodiments, the techniques described herein result in the avoidance of the physical transfer of cell culture fluid from one bioreactor to another, thus reducing the number of manual processes and increasing the potential for process automation.

[0200] In some embodiments, the techniques described herein provide efficiency improvements. For example, in some embodiments, automated processes (e.g., phase transitions caused by cell density (e.g., seeding, filling of the bioreactor, addition of inducers, etc.)) impose little or no cell waste, process waste, and / or time waste on the manufacturer. In some embodiments, the techniques described herein result in a reduction in labor / handling effort. For example, in some embodiments, since the collection system and the perfusion system are the same, there is no need to prepare a dedicated collection device to receive the cell culture fluid (e.g., culture broth) generated during the perfusion preparation process.

[0201] In some embodiments, the techniques described herein result in a reduction in the consumable consumption rate. For example, in some embodiments, a reduction of greater than or equal to about 50% of the required consumables can be achieved. In some embodiments, the techniques described herein result in a reduction in consumable costs because, for example, no additional cell retention devices (CRDs) are required for collection (e.g., using the same CRD between the perfusion stage and the collection stage), and one CRD can be used for both processes. In some embodiments, the techniques provided herein can include the use of the same CRD during a perfusion process that includes two or more perfusion stages, such as 1, 2, 3, 4, 5, 6, or more than 6 perfusion stages, including but not limited to these. In some embodiments, the perfusion process includes a series of perfusion stages (e.g., a seed train). In some embodiments, a series of perfusion stages can include a progressive increase in the volume of the bioreactor and / or the viable cell density at each subsequent stage. In some embodiments, the techniques provided herein can include the use of the same CRD for 1, 2, 3, 4, 5, 6, or more than 6 perfusion stages, and at least one collection stage for about 1, 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, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, or 70 or more days, or more than 70 days. Thus, in some embodiments, the techniques described herein result in less consumable storage management effort and / or a smaller storage footprint.

[0202] In some embodiments, the techniques described herein reduce the risk of contamination. For example, in some embodiments, there is less manual application, less transfer of cell culture fluid or biologic production materials, and a reduced likelihood of introduction of contaminants.

[0203] In some embodiments, the techniques described herein result in a reduction in the number of CRDs required during a biomanufacturing process, particularly under conditions of large-scale biomanufacturing. For example, in some embodiments, the CRDs utilized during a perfusion process can be utilized during a harvest process (e.g., batch, fed-batch, continuous, and / or semi-continuous harvest). In some embodiments, the CRDs (e.g., HF filters) are already coupled during the perfusion process and are reused during a harvest stage (e.g., perfusion, batch, supply batch, continuous, semi-continuous, bulk harvest, etc.). In some embodiments, the CRDs remain attached to the bioreactor during transitions between processes. In some embodiments, the CRDs are in a "standby" mode, e.g., cells are still circulating through the lumen of the filter fiber, but the permeate is not drawn through the CRD.

[0204] The following description provides exemplary embodiments of the apparatus, system, and method described herein for biomanufacturing. Explanations and examples of various terms used herein are provided in Section II below.

[0205] II. Examples of Term Explanations It should be understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting.

[0206] Unless otherwise defined, all technical terms, notations, and other scientific, technical, or specialized terms used in this specification are intended to have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined in this specification for clarity and / or ease of reference, and including such definitions in this specification should not necessarily be construed as representing a substantial difference from what is commonly understood in the art. Generally, the nomenclature and techniques utilized in connection with chemistry, biochemistry, molecular biology, pharmacology, and toxicology described herein are well known and commonly used in the art.

[0207] Throughout this disclosure, various aspects are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the disclosure. Accordingly, a recitation of a range should be considered to specifically disclose all the possible sub-ranges as well as the individual numerical values within that range. For example, when a range of values is provided, it is understood that each intervening value between the upper and lower limits of that range, as well as any other stated or intervening value within the stated range, is included in the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also included in the disclosure in accordance with any specifically excluded limits within the stated range. When the stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included in the disclosure. This applies regardless of the breadth of the range.

[0208] The headings and subheadings between sections and subsections of this document are included solely to improve readability and do not imply that features cannot be combined across sections and subsections. Accordingly, sections and subsections do not describe separate embodiments.

[0209] All references cited herein, including patent applications, patent publications, and UniProtKB / Swiss-Prot accession numbers, are hereby incorporated by reference in their entirety as if each individual reference were specifically and individually indicated to be incorporated by reference.

[0210] As used herein, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. The term "and / or" as used herein refers to any and all possible combinations of one or more of the associated listed items and is to be understood to encompass them. As used herein, the terms "includes", "including", "comprises", and / or "comprising" specify the presence of the stated features, integers, steps, operations, elements, components, and / or units, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, units, and / or groups thereof.

[0211] The term "about" as used herein refers to including the normal error range for each value readily known. References to values or parameters with "about" herein include (and describe) embodiments directed to the value or parameter itself. For example, a description referring to "about X" includes a description of "X". In some embodiments, "about" can refer to ±15%, ±10%, ±5%, or ±1% as understood by one of ordinary skill in the art.

[0212] The term "amino acid" as used herein generally refers to any organic compound containing an amino group (e.g., -NH2), a carboxyl group (-COOH), and a side chain group (R) that varies based on the particular amino acid. Amino acids can be linked using peptide bonds.

[0213] As used herein, the phrase "at least one" when used in conjunction with a list of items means that one or more different combinations of the listed items may be used and only one of the items in the list may be required. An item can be a particular object, thing, step, action, process, or category. In other words, "at least one of" means that any combination or any number of items from the list may be used, but not all of the items in the list are required. For example, without limitation, "at least one of item A, item B, or item C" or "at least one of item A, item B, and item C" can mean item A; item A and item B; item B; item A, item B, and item C; item B and item C; or item A and item C. In some cases, "at least one of item A, item B, or item C" or "at least one of item A, item B, and item C" may mean two of item A, one of item B, and ten of item C; four of item B and seven of item C; or some other suitable combination, but is not limited thereto.

[0214] The use of the term "or" in the claims is used to mean "and / or" unless explicitly indicated to refer to only alternatives or the alternatives are mutually exclusive, but the present disclosure supports a definition that refers to only alternatives and "and / or". For example, "x, y, and / or z" can refer to "x" alone, "y" alone, "z" alone, "x, y, and z", "(x and y) or z", "x or (y and z)", or "x or y or z". It is specifically contemplated that x, y, or z may be specifically excluded from an embodiment.

[0215] As used herein, the term "biological agent" generally refers to biological molecules such as proteins or chemical substances produced by cells or viruses. This term can mean, but is not limited to, proteins, peptides, antibodies (including, but not limited to, antibody derivatives such as Fc fusions, scFvs, multispecific antibodies, bispecific antibodies), nucleic acids, metabolites, antigens, chemical substances, or any molecule such as a biopharmaceutical. This term can also mean an adeno-associated virus (AAV), such as an AAV-based gene therapy vector produced by cells. In some embodiments, the biological agent is readily administrable and / or can be made so, suitable for administration to a subject. In some embodiments, the biological agent is purified and / or formulated before it is administered to a subject so as to be suitable for such administration.

[0216] As used herein, the terms "biological sample", "biological specimen", or "biological material" generally refer to a specimen that has been sampled, typically to represent the source of the specimen from a subject. A biological sample can represent the whole subject, a particular tissue, cell type, or organism as a category or subcategory. A biological sample can contain macromolecules. A biological sample can contain small molecules. A biological sample can contain viruses. A biological sample can contain cells or cell derivatives. A biological sample can contain organelles. A biological sample can contain cell nuclei. A biological sample can contain rare cells from a population of cells. A biological sample can be of any cell type, including, but not limited to, prokaryotic cells, eukaryotic cells, bacteria, fungi, plants, mammals, or other animal cell types, mycoplasma, normal tissue cells, tumor cells, or any other cell type, whether derived from a single cell or a multicellular organism. A biological sample can contain components of cells. A biological sample can contain nucleotides (e.g., ssDNA, dsDNA, RNA), organelles, amino acids, peptides, proteins, carbohydrates, glycoproteins, or any combination thereof. A biological sample can be or contain a matrix (e.g., a gel or polymer matrix) containing one or more components derived from cells (e.g., cell beads), such as DNA, RNA, organelles, proteins, or any combination thereof, from cells. A biological sample can be obtained from a tissue of a subject. A biological sample can contain fixed cells. Such fixed cells may or may not contain a cell wall or cell membrane. A biological sample can contain one or more components of a cell, but may not contain other components of the cell. Examples of such components can include nuclei or organelles. A biological sample can contain live cells. The live cells can be culturable.

[0217] Throughout this specification, unless the context requires otherwise, the words "comprise", "comprises" and "comprising" are to be interpreted as including the stated step or element or group of steps or elements but not excluding any other step or element or group of steps or elements. As used herein, "consisting of" means including and limited to what follows the phrase "consisting of". Thus, the phrase "consisting of" indicates that the listed elements are necessary or essential and that no other elements can exist. "Consisting essentially of" means including any elements listed after the phrase and limited to other elements that do not interfere with or contribute to the activity or action specified in this disclosure for the listed elements. Thus, the phrase "consisting essentially of" indicates that the listed elements are necessary or essential, but other elements are not optional and may or may not exist depending on whether they affect the activity or action of the listed elements.

[0218] In addition, when terms such as "coupled to", "communicates with" or "is communicatively associated with" or similar terms are used herein, one element may be capable of communicating directly, indirectly, or both, with another element by one or more wired communication links, one or more wireless communication links, one or more optical communication links, or combinations thereof. Further, when a list of elements (e.g., elements a, b, c) is referenced, such reference is intended to include any one of the elements listed therein by itself, any combination consisting of less than all of the elements listed therein, and / or any combination of all of the elements listed therein.

[0219] As used herein, "contaminant" can refer to anything that is considered undesirable in a cell culture fluid and / or cell culture medium. For example, under certain conditions, contaminants can include, but are not limited to, unintended cells and / or viruses, cell culture debris, cell products that are not the target product for therapeutic purposes, cell metabolites, etc.

[0220] As used herein, "internal standard" can refer to something that can be included in the same sample as the target (e.g., spiked in). The internal standard can be used for calibration purposes. Further, the internal standard can be used in the systems and methods described herein.

[0221] Throughout this specification, references to "one embodiment", "an embodiment", "a particular embodiment", "related embodiments", "a particular embodiment", "additional embodiments", or "further embodiments", or combinations thereof, mean that the particular features, structures, or characteristics described in connection with the embodiment are included in at least one embodiment of the invention. Thus, the appearances of the foregoing phrases in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0222] As used herein, "feed stream" is the fluid stream exiting the bioreactor, which can include cells, cell culture medium, biopharmaceuticals, etc. and is directed to a filter. Generally, the feed stream contains a lower concentration of cells than the hold-up stream. In some cases, when the permeate is not being withdrawn from the filter, both streams can have the same concentration of cells.

[0223] As used herein, the term "collected permeate stream" includes the fluid that passes through the filter membrane and exits the filter, which can include cell culture medium, biopharmaceuticals, etc. and is generally targeted for further processing. Generally, the collected permeate stream is active during the collection process and contains a higher concentration of biopharmaceuticals and a lower concentration of cells than the hold-up stream.

[0224] As used herein, "model" may include one or more algorithms, one or more mathematical methods, one or more machine learning algorithms, or combinations thereof.

[0225] As used herein, the term "multi-purpose assembly" generally refers to a device that includes at least two flow paths, each flow path being operably connected to a pump configured to draw fluid from the permeate flow of the cell retention device at a desired rate.

[0226] As used herein, the term "ones" means more than one.

[0227] As used herein, the term "plurality" can be 2, 3, 4, 5, 6, 7, 8, 9, 10 or more.

[0228] As used herein, the term "peptide" generally refers to amino acids linked by peptide bonds. A peptide can include an amino acid chain between 10 and 50 residues. A peptide can include an amino acid chain shorter than 10 residues, including oligopeptides, dipeptides, tripeptides, and tetrapeptides. A peptide can include a chain longer than 50 residues and may be referred to as a "polypeptide" or "protein".

[0229] As used herein, the term "perfusion permeate flow" includes the fluid that passes through the filter membrane and exits the filter, may include cell culture medium, biopharmaceuticals, etc., and is generally the subject of further processing. Generally, the perfusion permeate flow is active during the perfusion process and includes a higher concentration of cell culture medium and a lower concentration of biopharmaceuticals than the collected permeate flow.

[0230] As used herein, the term "permeate" includes the fluid that passes through the filter membrane and exits the filter.

[0231] The terms "protein", "polypeptide", or "peptide" may be used interchangeably herein and generally refer to a molecule containing at least three amino acid residues. A protein can comprise a polymeric chain made from an amino acid sequence linked together by peptide bonds. A protein can be digested in preparation for mass spectrometry using a trypsin digestion protocol. If access is limited to the cleavage site, other proteases can be used to digest the protein in preparation for mass spectrometry.

[0232] As used herein, the term "retentate flow" includes the fluid exiting the filter without passing through the filter membrane (e.g., passing through the filter lumen), which can include cells, cell culture medium, biologic agents, etc., and is generally directed to a bioreactor. Generally, the retentate flow contains a higher concentration of cells than the feed flow. In some cases, when the permeate is not being withdrawn from the filter, both flows can have the same concentration of cells.

[0233] As used herein, the term "sample" generally refers to a sample obtained from a subject of interest and may include a biological sample of the subject. The sample may include a cell sample. The sample may include a cell line or a cell culture sample. The sample can contain one or more cells. The sample can contain one or more microorganisms. The sample may include a nucleic acid sample or a protein sample. The sample may also include a carbohydrate sample or a lipid sample. The sample may be derived from another sample. The sample may include a tissue sample such as a biopsy, a core biopsy, a needle aspirate, or a fine needle aspirate. The sample may include a fluid sample such as a blood sample, a urine sample, or a saliva sample. The sample may include a skin sample. The sample may include a buccal swab. The sample may include a plasma or serum sample. The sample may include a cell-free sample. The cell-free sample may contain extracellular polynucleotides. The sample may be derived from blood, plasma, serum, urine, saliva, mucosal excretions, sputum, feces, or tears. The sample may be derived from red blood cells or white blood cells. The sample may be derived from feces, cerebrospinal fluid, CNS fluid, gastric fluid, amniotic fluid, cyst fluid, peritoneal fluid, bone marrow, bile, other body fluids, tissue obtained from a biopsy, skin, or hair.

[0234] As used herein, the term "set of" means one or more. For example, a set of items includes one or more items.

[0235] As used herein, the term "sequence" generally refers to a biological sequence that includes one-dimensional monomers that can be assembled to generate a polymer. Non-limiting examples of sequences include nucleotide sequences (e.g., ssDNA, dsDNA, and RNA), amino acid sequences (e.g., proteins, peptides, and polypeptides), and carbohydrates (e.g., compounds containing C m (H2O) n and compounds containing).

[0236] As used herein, the term "subject" generally refers to an animal such as a mammal (e.g., human) or a bird (e.g., avian), or another organism such as a plant. For example, a subject can include a vertebrate, a mammal, a rodent (e.g., mouse), a primate, a monkey, or a human. Animals can include, but are not limited to, livestock, sport animals, and pets. A subject can include a healthy or asymptomatic individual, an individual having or suspected of having a disease (e.g., cancer) or a predisposition to a disease, and / or an individual in need of or suspected of being in need of treatment. A "subject" can be a "patient". A subject can include a microorganism or a pathogen (e.g., a bacterium, a fungus, an archaeon, a virus).

[0237] As used herein, "substantially" means being sufficient to function for the intended purpose. Thus, the term "substantially" allows for minor, insignificant variations from an absolute or complete state, dimension, measurement, result, etc., that are expected by one of ordinary skill in the art but do not materially affect overall performance. When used with respect to a numerical or parameter or a property that can be expressed as a numerical value, "substantially" means within 10 percent.

[0238] The following description provides only exemplary embodiments and is not intended to limit the scope, applicability, or configuration of the present disclosure. Rather, the following description of the preferred exemplary embodiments provides possible explanations to those of ordinary skill in the art for implementing various embodiments. It is understood that the functions and arrangements of the elements can be varied in various ways without departing from the spirit and scope as set forth in the appended claims.

[0239] In the following description, specific details are provided to give a comprehensive understanding of the embodiments. However, it will be understood that the embodiments may be practiced without these specific details. For example, circuits, systems, networks, processes, and other components may be shown as components in block diagram form in order not to obscure the embodiments with unnecessary detail. In other instances, well-known circuits, processes, algorithms, structures, and techniques may be shown without unnecessary detail in order to avoid obscuring the embodiments.

[0240] III. Overview of the Multi-Purpose Assembly Referring to FIG. 1, a non-limiting schematic view of an exemplary multi-purpose assembly 100 for the improved production of biopharmaceuticals is shown. The assembly comprises a supply flow conduit 101 operably connected to the inlet of a cell retention device such as a hollow fiber (HF) filter 102. In certain embodiments, the supply flow conduit is operable with any supply pump 121. In some embodiments, the HF filter is operable with tangential flow. In some embodiments, the HF filter is a tangential flow depth filtration (registered trademark) (TFDF (registered trademark)) filter. The assembly is operable to facilitate the entry of fluid (e.g., a fluid containing cell culture medium, cells, biopharmaceuticals, etc.) into the HF filter while allowing suitable fluid components (e.g., those of a size sufficient to do so) to pass through the filtration membrane and exit the HF filter as permeate 103. The HF filter is operable such that fluid that does not pass through the filter membrane can exit the HF filter as retentate 112. The permeate fluid can be drawn through the HF filter by the action of two or more pumps and enter two or more flow paths. Under certain operating conditions, such as operating in a perfusion process, the first pump 106 is operable to draw the permeate 103 into the first flow path 104 to form a perfusion permeate 107. The first pump 106 is operable to maintain a fluid flow rate suitable for a perfusion process, for example, such that the perfusion permeate 107 from the HF filter is pulled at a consistent and / or desirable rate, and is operably in communication with a sensor 105 operable for the accurate measurement of fluid flow rate. Under certain operating conditions, such as operating in a collection process, the second pump 110 is operable to draw the permeate 103 into the second flow path 108 to form a collection permeate 111. The second pump 110 and the collection permeate 111 are optionally operable to communicate with one or more sensors and / or to be in fluid communication with one or more downstream filtration units and / or processing units suitable for maintaining the collection process. Generally, the second flow path 108 and the second pump 110 draw the permeate at a higher fluid flow rate than the first flow path 104 and the first pump 106.

[0241] In certain embodiments, the multi-purpose assembly is operable for one or more perfusion processes (e.g., but not limited to, 1, 2, 3, 4, 5, 6, or more than 6 perfusion processes) and one or more collection processes (e.g., a collection process and / or additional processes after the start of a collection process). In certain embodiments, the multi-purpose assembly is operable for one or more perfusion processes, one or more intermediate processes (e.g., a process between one or more processes, in such a process, the permeate may or may not be withdrawn, and the fluid flow may or may not be maintained), and one or more collection processes (e.g., additional processes after the start of a collection process). In some embodiments, such a combination of processes is referred to as a combined process.

[0242] In certain embodiments, the multi-purpose assembly is connected to a turndown bioreactor. In certain embodiments, the turndown ratio is from about 30:1 to about 2:1. In certain embodiments, the turndown ratio is from about 20:1 to about 5:1. In certain embodiments, the turndown ratio is from about 10:1 to about 5:1. In certain embodiments, one or more of the multi-purpose assemblies used in the methods of the present disclosure are connected to a bioreactor having a turndown ratio of about 10:1. In certain embodiments, one or more of the multi-purpose assemblies used in the methods of the present disclosure are connected to a bioreactor having a turndown ratio of about 5:1.

[0243] In certain embodiments, two or more multi-purpose assemblies can be utilized in one system. In certain embodiments, one, two, three, four, or five multi-purpose assemblies are utilized in one system. In certain embodiments, when two or more multi-purpose assemblies are utilized within one system, the assemblies are utilized sequentially and / or in tandem. In certain embodiments, the utilization of two or more multi-purpose assemblies in a system that includes a bioreactor can enable an increase in the volume of the bioreactor as compared to a system having only one multi-purpose assembly.

[0244] In certain embodiments, exemplary operating parameters of throughput for three target outputs achievable using a multi-purpose assembly and / or a system comprising the same are shown in Table 1. [Table 1]

[0245] III.A. Cell Retention Device (CRD) In certain embodiments, the CRD is a filter suitable for both perfusion and collection processes. In certain embodiments, the CRD is a hollow fiber (HF) filter. In certain embodiments, the HF filter is operable with respect to tangential flow. In certain embodiments, the HF filter is a 3E-NF20A, 3E-NF40A, 3E-NF60, 3E-NF80A, 3E-NF90A (3E Memtech Pte Ltd.), De.mem NF (De.mem Limited), dNF80, dNF40 (NX Filtration), HFW100 (Pentair X-Flow (trademark)), NUF N80 (Ochemate (registered trademark)), or a tangential flow depth filtration (registered trademark) (TFDF (registered trademark); Repligen (registered trademark) Inc.) filter. In certain embodiments, the HF filter is a tangential flow depth filtration (registered trademark) (TFDF (registered trademark)) filter. In certain embodiments, the HF filter is composed of and / or includes PES / PVDF, PEI-based TFC, PES-PEM, mPES, and / or polyamide-based TFC.

[0246] In some embodiments, the HF filter is constructed of and / or includes a synthetic polymer. In some embodiments, the HF filter is composed of and / or includes polypropylene and / or polyethylene terephthalate. In some embodiments, the HF filter is constructed of and / or includes modified polyethersulfone, polyethersulfone, mixed cellulose ester, and / or polysulfone.

[0247] In certain embodiments, the HF filter has a filter surface area in the range of about 3 to about 6240 cm 2 . In certain embodiments, the HF filter is about 3 cm 2 , about 30 cm 2 , about 50 cm 2 , about 150 cm 2 , about 450 cm 2 , about 750 cm 2 , about 1,500 cm 2 , about 2,100 cm2 and having a filter surface area of about 6,000 cm 2 or greater than about 6,000 cm 2 . In certain embodiments, the HF filter has a filter surface area of about 150, 300, 450, 600, 750, 900, 1,050, 1,200, 1,350, 1,500, 1,650, 1,800, 1,950, 2,100, 2,250, 2,400, 2,550, 2,700, 2,850, 3,000, 3,150, 3,300, 3,450, 3,600, 3,750, 3,900, 4,050, 4,200, 4,350, 4,500, 4,650, 4,800, 4,950, 5,100, 5,250, 5,400, 5,550, 5,700, 5,850, 6,000, 6,150, 6,300, 6,450, 6,600, 6,750, 6,900, 7,050, 7,200, 7,350, 7,500, 7,650, 7,800, 7,950, 8,100, 8,250, 8,400, 8,550, 8,700, 8,850, 9,000, or greater than 9,000 cm 2 .

[0248] In certain embodiments, the HF filter includes one or more fibers. In certain embodiments, the fiber has a lumen through which a feed stream can flow, surrounded by a membrane layer through which a permeate can pass. In certain embodiments, the permeate fluid withdrawn through the membrane layer can leave the HF filter as the permeate (see, e.g., permeate 103 of FIG. 1). In certain embodiments, the tube is 20, 36, or 108 cm in length. In certain embodiments, the tube is 1, 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, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, or greater than 130 cm in length.

[0249] In certain embodiments, the HF filter comprises two or more fibers. In some embodiments, the HF filter comprises 1, 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, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 or more fibers.

[0250] In some embodiments, the HF filter has an average pore size of from about 0.65 μm to about 8 μm. In some embodiments, the HF filter has an average pore size of from about 2 μm to about 5 μm. In some embodiments, the HF filter has an average pore size of about 2 μm. In certain embodiments, the HF filter can have an average pore size of about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 μm, or any derivable range therein. In certain embodiments, the pore structure is anisotropic. In certain embodiments, the pore structure is not anisotropic. In certain embodiments, the pore structure is isotropic.

[0251] In certain embodiments, the HF filter filtrate channel includes a filter having a lumen with an inner diameter of about 2 mm therethrough. In certain embodiments, the HF filter filtrate channel includes a filter having a lumen with an inner diameter of about 4.6 mm therethrough. In certain embodiments, the TFDF® filtration filtrate channel includes a filter having an inner diameter lumen of about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 mm, or any range derivable therefrom. In certain embodiments, the HF filter filtrate channel includes a filter having a lumen from about 0.4 mm to about 1.5 mm. In certain embodiments, the HF filter filtrate channel does not have a lumen with an inner diameter of less than about 0.4 mm therethrough.

[0252] In certain embodiments, the HF filter has a wall thickness from about 0.05 mm to 0.075, 0.1, 0.125, 0.15, 0.175, 0.2, 0.225, 0.25, 0.275, 0.3, 0.325, 0.35, 0.375, 0.4, 0.425, 0.45, 0.475, 0.5 mm, or any range derivable therein. In certain embodiments, the HF filter has a wall thickness from about 1 mm to about 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5 mm, or any range derivable therein. In certain embodiments, the HF filter has a wall thickness of about 5 mm.

[0253] In certain embodiments, the permeate can flow through the HF filter at a flow rate greater than about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17.0, 17.5, 18.0, 18.5, 19.0, 19.5, or 20.0 liters per minute (LPM). In certain embodiments, the permeate can flow through the HF filter at a flow rate between about 0 LPM and / or about 0.1 LPM to about 20 LPM, or any range derivable therefrom. In certain embodiments, the permeate can flow through the HF filter at a flow rate in the range between about 0 LPM and / or about 0.1 LPM to about 5 LPM. In certain embodiments, the permeate can flow through the HF filter at a flow rate in the range between about 0 LPM and / or about 0.1 LPM to about 8 LPM. In certain embodiments, the permeate can flow through the HF filter at a flow rate in the range between about 0 LPM and / or about 0.1 LPM to about 10 LPM. In certain embodiments, the permeate can flow through the HF filter at a flow rate in the range between about 0 LPM and / or about 0.1 LPM to about 13 LPM. In certain embodiments, the permeate can flow through the HF filter at a flow rate in the range between about 0 LPM and / or about 0.1 LPM to about 15 LPM. In certain embodiments, the permeate can flow through the HF filter at a flow rate in the range between about 0 LPM and / or about 0.1 LPM to about 18 LPM.

[0254] In certain embodiments, the permeate can flow through the HF filter while maintaining a shear rate of less than about 10,000, 9,000, 8,000, 7,000, 6,000, 5,000, 4,500, 4,000, 3,500, 3,000, 2,500, 2,000, 1,500, 1,000, or 500 s-1.

[0255] In certain embodiments, the HF filter has a throughput greater than about 1,000, 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, 10,000, 11,000, 12,000, 13,000, 14,000, 15,000, 16,000, 17,000, 18,000, 19,000, 20,000, 21,000, 22,000, 23,000, 24,000, 25,000, 26,000, 27,000, 28,000, 29,000, 30,000, 31,000, 32,000, 33,000, 34,000, 35,000, 36,000, 37,000, 38,000, 39,000, 40,000, 41,000, 42,000, 43,000, 44,000, 45,000, 46,000, 47,000, 48,000, 49,000, 50,000, 51,000, 52,000, 53,000, 54,000, 55,000, 56,000, 57,000, 58,000, 59,000, 60,000, 61,000, 62,000, 63,000, 64,000, 65,000, 66,000, 67,000, 68,000, 69,000, 70,000, 71,000, 72,000, 73,000, 74,000, 75,000, 76,000, 77,000, 78,000, 79,000, or 80,000 liters per square meter (L / M 2 ). In some embodiments, the HF filter has a throughput greater than about 10,000 L / m 2 . In some embodiments, the HF filter has a throughput greater than about 25,000 L / m 2 . In some embodiments, the HF filter has a throughput greater than about 30,000 L / m 2 . In some embodiments, the HF filter has a throughput greater than about 40,000 L / m 2 . In some embodiments, the HF filter has a throughput equal to about 5,000 to about 35,000 L / m 2 , or any range derivable therefrom.

[0256] In certain embodiments, the HF filter has a filter flow rate of from about 20 to about 800 liters per square meter per hour (LMH). In certain embodiments, the HF filter has a filter flow rate of from about 100 to about 600 LMH. In certain embodiments, the HF filter has a filter flow rate of from about 300 to about 600 LMH. In certain embodiments, the HF filter has a filter flow rate of from about 400 to about 600 LMH. In certain embodiments, the HF filter has a filter flow rate of from about 50 to about 100, from about 50 to about 150, from about 50 to about 200, from about 50 to about 250, from about 50 to about 300, from about 50 to about 350, from about 50 to about 400, from about 50 to about 450, from about 50 to about 500, from about 50 to about 550, from about 50 to about 600, from about 50 to about 650, from about 50 to about 700, from about 50 to about 750, from about 50 to about 800, from about 50 to about 850, or from about 50 to about 900 LMH.

[0257] In some embodiments, the multi-purpose filter assembly is operable to maintain a viable cell density (VCD) greater than about 1×10 6 cells / mL. In some embodiments, the multi-purpose filter assembly is operable to maintain a viable cell density (VCD) greater than about 10×10 6 cells / mL. In some embodiments, the multi-purpose filter assembly is operable to maintain a viable cell density (VCD) greater than about 25×10 6 cells / mL. In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD greater than about 50×10 6 cells / mL. In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD greater than about 75×10 6 cells / mL. In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD greater than about 10×10 7 cells / mL. In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD greater than about 25×10 7 cells / mL. In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD greater than about 50×10 7It is operable to maintain a VCD exceeding cells / mL. In some embodiments, the multi-purpose filter assembly is about 75×10 7 It is operable to maintain a VCD exceeding cells / mL. In some embodiments, the multi-purpose filter assembly is about 10×10 8 It is operable to maintain a VCD exceeding cells / mL. In some embodiments, the multi-purpose filter assembly is about 25×10 8 It is operable to maintain a VCD exceeding cells / mL. In some embodiments, the multi-purpose filter assembly is about 50×10 8 It is operable to maintain a VCD exceeding cells / mL. In some embodiments, the VCD is the VCD in the bioreactor fluid volume. In some embodiments, the VCD is the VCD of the holding liquid flow of the CRD.

[0258] III.B. Pump In some embodiments, the assemblies, systems, and / or methods described herein include the use of one or more pumps.

[0259] In certain embodiments, the pump may be operable to push or pull fluid through a suitable conduit. In certain embodiments, the pump can be a pump selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetically driven pumps, positive displacement pumps, membrane pumps, pressure pumps, Quantex (e.g., positive displacement rotary pumps) pumps, gear pumps, diaphragm pumps, syringe pumps, and piston pumps, but is not limited thereto. In certain embodiments, the pump is a magnetically driven pump. In certain embodiments, the circulation pump is a peristaltic pump. In certain embodiments, one or more pumps are suitable for generating an alternating tangential flow. In certain embodiments, one or more pumps are not utilized and / or operable for generating an alternating tangential flow.

[0260] In certain embodiments, the pump is operable to maintain a desired flow rate. In certain embodiments, the pump communicates with one or more sensors that can be instructed to increase or decrease the pumping force to the pump to change the flow rate as needed. In some embodiments, the flow rate is measured in liters per fiber per minute (L / fiber / min). In some embodiments, the flow rate is 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5 L / fiber / min or approximately those values. In some embodiments, the flow rate is from about 0.5 to about 2.5 L / fiber / min. In some embodiments, the flow rate is from about 0.8 to about 2.2 L / fiber / min. In some embodiments, the flow rate is from about 0.8 to about 2.0 L / fiber / min. In some embodiments, the flow rate is from about 1 to about 1.8 L / fiber / min.

[0261] In some embodiments, the flow rate is determined as a function of a set volume per day (VVD) of the container. In some embodiments, the pump can maintain a flow rate equal to about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5 VVD. In some embodiments, the pump can maintain a flow rate of about 2 VVD. In some embodiments, the pump can maintain a flow rate of about 3 VVD. In some embodiments, the pump can maintain a flow rate of about 4 VVD. In some embodiments, the pump can maintain a flow rate of about 5 VVD. In some embodiments, the pump can maintain a flow rate greater than about 5 VVD. In some embodiments, when a relatively large filter is utilized with a relatively small bioreactor, the pump can maintain a higher VVD rate.

[0262] In some embodiments, the system can include two or more multipurpose assemblies. In some embodiments, where the system includes two or more multipurpose assemblies, accordingly, for example, but not limited to, the VVD rate can be amplified at about 5, 6, 7, 8, 9, 10, or greater than 10 VVD.

[0263] In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate exceeding about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17.0, 17.5, 18.0, 18.5, 19.0, 19.5, or 20.0 liters per minute (LPM). In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate exceeding about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17.0, 17.5, 18.0, 18.5, 19.0, 19.5, 20.0, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30, 30.5, 31, 31.5, 32, 32.5, 33, 33.5, 34, 34.5, 35, 35.5, 36, 36.5, 37, 37.5, 38, 38.5, 39, 39.5, 40, 40.5, 41, 41.5, 42, 42.5, 43, 43.5, 44, 44.5, 45, 45.5, 46, 46.5, 47, 47.5, 48, 48.5, 49, 49.5, 50, 50.5, 51, 51.5, 52, 52.5, 53, 53.5, 54, 54.5, 55, 55.5, 56, 56.5, 57, 57.5, 58, 58.5, 59, 59.5, 60, 60.5, 61, 61.5, 62, 62.5, 63, 63.5, 64, 64.5, 65, 65.5, 66, 66.5, 67, 67.5, 68, 68.5, 69, 69.5, 70, 70.5, 71, 71.5, 72, 72.5, 73, 73.5, 74, 74.5, 75, 75.5, 76, 76.5, 77, 77.5, 78, 78.5, 79, 79.5, or 80 LPM, or any range derivable therein.

[0264] In some embodiments, the pump is operable to accurately facilitate fluid flow at a flow rate in the range of from about 0 LPM and / or about 0.1 LPM to about 20 LPM, or any range derivable therefrom. In some embodiments, the pump is operable to accurately facilitate fluid flow at a flow rate in the range of from about 0 LPM and / or about 0.1 LPM to about 5 LPM. In some embodiments, the pump is operable to accurately facilitate fluid flow at a flow rate in the range of from about 0 LPM and / or about 0.1 LPM to about 8 LPM. In some embodiments, the pump is operable to accurately facilitate fluid flow at a flow rate in the range of from about 0 LPM and / or about 0.1 LPM to about 10 LPM. In some embodiments, the pump is operable to accurately facilitate fluid flow at a flow rate in the range of from about 0 LPM and / or about 0.1 LPM to about 13 LPM. In some embodiments, the pump is operable to accurately facilitate fluid flow at a flow rate in the range of from about 0 LPM and / or about 0.1 LPM to about 15 LPM. In some embodiments, the pump is operable to accurately facilitate fluid flow at a flow rate in the range of from about 0 LPM and / or about 0.1 LPM to about 18 LPM.

[0265] In some embodiments, the first flow path pump is operable to accurately facilitate fluid flow at a rate less than that of the second flow path pump. In some embodiments, the pump associated with the perfusion permeate flow path is operable to accurately facilitate fluid flow at a rate less than that of the pump associated with the collection permeate flow path.

[0266] In some embodiments, the first flow path pump associated with an accurate sensor is suitable for accurate control of the fluid flow rate in the first flow path. In some embodiments, the first flow path pump associated with an accurate sensor is suitable for accurate control of the fluid flow rate in the first flow path, and the control is more accurate than the pump utilized to direct the flow into the second flow path.

[0267] In some embodiments, the pump may have a passageway of any size. In some embodiments, the passageway of the pump is sized according to the desired flow rate, tube inner diameter, and / or flow accuracy requirements. In some embodiments, the tube of the flow path can be of any type suitable for culturing cells. In some embodiments, the tube can be autoclavable. In some embodiments, the tube is suitable for food grade quality and / or good manufacturing practice quality. In some embodiments, the tube includes silicone.

[0268] In some embodiments, the pump is operable to maintain a percentage of packed cell volume (PCV) of about 1, 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, or 50%, or any range derivable therefrom in blood. In some embodiments, the pump is operable to maintain a PCV percentage of about 2 to about 40%, about 8 to about 40%, about 12 to about 35%, or about 15 to about 30%.

[0269] III.C. Sensor The multi-purpose assembly includes at least one sensor suitable for accurately measuring the fluid flow rate of a first flow path (see, e.g., flow path 104 in FIG. 1). In some embodiments, such a sensor is in communication with a first flow path pump. In some embodiments, such a sensor is in communication with a second flow path pump.

[0270] In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate greater than about 0.01, 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17.0, 17.5, 18.0, 18.5, 19.0, 19.5, or 20.0 liters per minute (LPM).

[0271] In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate in the range of about 0 LPM and / or about 0.1 LPM to about 20 LPM, or any range derivable therefrom. In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate in the range of about 0 LPM and / or about 0.1 LPM to about 5 LPM. In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate in the range of about 0 LPM and / or about 0.1 LPM to about 8 LPM. In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate in the range of about 0 LPM and / or about 0.1 LPM to about 10 LPM. In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate in the range of about 0 LPM and / or about 0.1 LPM to about 13 LPM. In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate in the range of about 0 LPM and / or about 0.1 LPM to about 15 LPM. In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate in the range of about 0 LPM and / or about 0.1 LPM to about 18 LPM.

[0272] In some embodiments, at least one sensor suitable for accurately measuring the fluid flow rate of the first flow path is more accurate than one or more additional sensors utilized to measure the flow rate of the second flow path.

[0273] In some embodiments, the sensor is operable to communicate with a transmitter. In some embodiments, the transmitter is operable to communicate with one or more pumps, one or more control units, and / or one or more human-machine interfaces.

[0274] III.D. Flow Paths As described herein, in most embodiments, the multi-purpose assembly includes at least two flow paths. In certain embodiments, the multi-purpose assembly includes two flow paths. In certain embodiments, the multi-purpose assembly includes more than two flow paths, such as, but not limited to, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10 flow paths, or any derivable range therein. In some embodiments, in addition to the first two flow paths, the flow paths can be specialized to be operable for a particular process (e.g., a particular perfusion and / or collection process) or generalized to be operable for any process for which a flow path is required.

[0275] In certain embodiments, the first flow path is a flow path for perfusion permeate (see, e.g., FIG. 1, first flow path 104 and perfusion permeate 107). In some embodiments, the first flow path includes a tube having an inner diameter (ID) of 1 / 2 inch, 3 / 8 inch, 1 / 4 inch, or 1 / 8 inch. In certain embodiments, the first flow path includes a 3 / 8 inch ID tube.

[0276] In certain embodiments, the second flow path is a flow path for collecting permeate (see, e.g., FIG. 1, flow path 108, and collected permeate 111). In some embodiments, the second flow path includes a tube having an inner diameter (ID) of 1 / 2 inch, 3 / 8 inch, 1 / 4 inch, or 1 / 8 inch. In certain embodiments, the second flow path includes a 1 / 2 inch ID tube.

[0277] In certain embodiments, a multi-purpose assembly comprising a first flow path and a pump is operable to draw a flow rate from an HF filter at about 0 to about 3 liters per fiber per minute (L / fiber / min). In some embodiments, the first flow path and the pump are operable to draw a flow rate of about 0.8 to about 2.2 L / fiber / min from the HF filter. In some embodiments, the first flow path and the pump are operable to draw a flow rate of about 0.8 to about 2.0 L / fiber / min from the HF filter. In some embodiments, the first flow path and the pump are operable to draw a flow rate of about 1.0 to about 1.8 L / fiber / min from the HF filter.

[0278] In certain embodiments, a multi-purpose assembly comprising a second flow path and a pump is operable to draw a flow rate from an HF filter at about 0 to about 4 liters per fiber per minute (L / fiber / min). In some embodiments, the second flow path and the pump are operable to draw a flow rate of about 1 to about 2.5 L / fiber / min from the HF filter. In some embodiments, the second flow path and the pump are operable to draw a flow rate of about 1 to about 3.0 L / fiber / min from the HF filter. In some embodiments, the second flow path and the pump are operable to draw a flow rate of about 1.5 to about 2 L / fiber / min from the HF filter.

[0279] In some embodiments, a multi-purpose assembly comprising a second flow path and a pump is operable to draw fluid from an HF filter at a flow rate of about 0.01 LPM to about 80 LPM, about 0.01 LPM to about 60 LPM, about 0.01 LPM to about 40 LPM, and / or about 0.01 LPM to about 20 LPM.

[0280] In some embodiments, a multi-purpose assembly comprising a second flow path and a pump is about 1,000 liters per square meter (L / m 2 ) to about 80,000 L / m 2 , about 10,000 liters per square meter (L / m 2 ) to about 50,000 L / m 2, and / or about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 and is operable to draw fluid from the HF filter at a throughput of or greater. In some embodiments, the multi-purpose assembly comprising the second flow path and the pump is operable to draw fluid from the HF filter at a throughput of about 4,000 liters per square meter (L / m 2 ) or greater. In some embodiments, the multi-purpose assembly comprising the second flow path and the pump is operable to draw fluid from the HF filter at a throughput of about 40,000 liters per square meter (L / m 2 ) or greater.

[0281] In some embodiments, the first flow path has an accuracy requirement of about 1%. In some embodiments, the first flow path has an accuracy requirement of about 2%. In some embodiments, the second flow path has an accuracy requirement of about 3%. In some embodiments, the second flow path has an accuracy requirement of about 4%. The accuracy requirement can be an accuracy requirement of the flow.

[0282] In some embodiments, the first and second flow paths are joined to a permeate flow path (see, e.g., FIG. 1, flow path 103, first flow path 104, and second flow path 108). In some embodiments, the first and second flow paths of the multi-purpose assembly are connected by any suitable means such as, but not limited to, a T-connector, a Y-connector, or a valve. In some embodiments, any clamp (e.g., a pinch clamp) can be operable to direct the flow into the first or second flow path. In some embodiments, the opening and closing of the valve can be automated.

[0283] In some embodiments, the fluid flow rate through the first flow path is lower than the fluid flow rate through the second flow path. In some embodiments, the fluid flow rate through the perfusion flow path is less than the fluid flow rate through the collection flow path.

[0284] III.E. Filter In some embodiments, the multi-purpose assembly can include one or more additional filters (e.g., clarification filters). In some embodiments, the additional filter is attached to the first flow path. In some embodiments, the additional filter is attached to the second flow path. The additional filter can be configured to remove contaminants, selectively remove and / or collect one or more biologic agents, and / or ensure the sterility of the closed system, among other things.

[0285] III.F. Control Unit and Human Machine Interface In some embodiments, provided herein is a multi-purpose assembly that can be operably connected to at least one data processor, at least one control unit, and / or at least one human machine interface (HMI). In some embodiments, the control unit and HMI are used to monitor and / or control any adjustable parameters associated with the multi-purpose assembly, such as, but not limited to, VVD, flow rate, VCD, pump flow rate, sensor accuracy, use of flow paths, etc. In some embodiments, the control unit and HMI are used to control additional parameters associated with a system that includes the multi-purpose assembly, such as, but not limited to, temperature, fresh media feed rate, bleed rate, rate of addition of inductor agent, oxygen level, CO2 level, metabolite level, etc.

[0286] In some embodiments, a customized control strategy is programmed, integrated, and / or displayed using the HMI. In some embodiments, any physical component and / or device described herein is operable with respect to and / or can communicate with the HMI.

[0287] IV. Overview of Systems that Include a Multi-Purpose Assembly Referring to FIG. 2, a non-limiting schematic view of an exemplary bioreactor system 200 is shown that includes a multipurpose assembly (e.g., see FIG. 1, 100) for improved production of a biologic. The system comprises a bioreactor 220 operably connected to any inlet port (e.g., fresh media input port) 223, an output port (e.g., feed stream) 201, and an input (e.g., hold-up liquid) port 212. The system can include a bioreactor fluid volume 222. The system comprises a feed stream conduit 201 operably connected to the output port 201 and an inlet of a cell retention device such as a hollow fiber (HF) filter 202. In some embodiments, such HF filters are operable with respect to tangential flow. In some embodiments, such HF filters are tangential flow depth filtration (TM) (TFDF (TM)) filters. The output port 201 can optionally include a pump 221 to facilitate the flow of fluid from the bioreactor to the HF filter 202. The system is operable to facilitate the entry of fluid (e.g., fluid containing cell culture medium, cells, biologics, etc.) into the HF filter while allowing appropriate fluid components (e.g., those of a size sufficient to do so) to pass through the filtration membrane and exit the HF filter as permeate 203. The HF filter is operable such that fluid that does not pass through the filter membrane can exit the HF filter through an outlet as hold-up liquid 212 that is reintroduced into the bioreactor 220. The permeate fluid can be drawn through the HF filter by the action of two or more pumps and enter two or more flow paths. Under certain operating conditions such as operating in a perfusion process, the first pump 206 is operable to draw the permeate 203 into the first flow path 204 to form a perfusion permeate 207. The first pump 206 is operably in communication with a sensor 205 operable for accurate measurement of fluid flow rate such that, for example, the first pump 206 is operable to maintain a fluid flow rate suitable for a perfusion process in which the perfusion permeate 207 from the HF filter is drawn at a consistent and / or desirable rate.Under certain operating conditions, such as operating in the collection process, the second pump 210 is operable to draw the permeate 203 into the second flow path 208 to form a collected permeate 211. The second pump 210 and the collected permeate 211 are optionally operable to communicate with one or more sensors and / or to be in fluid communication with one or more downstream filtration units and / or processing units suitable for maintaining the collection process. Generally, the second flow path 208 and the second pump 210 draw the permeate at a higher fluid flow rate than the first flow path 204 and the first pump 206.

[0288] IV.A. Bioreactor In some embodiments, systems are described herein that include a multi-purpose assembly. In some embodiments, such systems are operable with a bioreactor. A bioreactor is a vessel operable to maintain a biologically active environment. The bioreactor may be suitable for maintaining either aerobic and / or anaerobic biological activity. In some embodiments, the bioreactor may be a batch bioreactor, a fed-batch bioreactor, a continuous bioreactor, a semi-continuous bioreactor, or a perfusion bioreactor. In some embodiments, the bioreactor can be of any size. In some embodiments, the bioreactor may be suitable for cell growth and / or maintenance during any stage of cell culture (e.g., induction phase, logarithmic phase, stationary phase, death phase, etc.). In some embodiments, the bioreactor may be made of any suitable material (e.g., stainless steel, etc.). In some embodiments, the bioreactor can have one or more input ports and / or one or more output ports. In some embodiments, the bioreactor may be operable to connect with two or more multi-purpose filter assemblies described herein. In some embodiments, where the bioreactor is operable with two or more multi-purpose filter assemblies, such two or more multi-purpose filter assemblies can function simultaneously or sequentially. In some embodiments, two or more bioreactor vessels can be utilized in tandem or continuously.

[0289] In some embodiments, the bioreactor vessel can include a volume of about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,100, 2,200, 2,300, 2,400, 2,500, 2,600, 2,700, 2,800, 2,900, 3,000, 3,100, 3,200, 3,300, 3,400, 3,500, 3,600, 3,700, 3,800, 3,900, 4,000, 4,100, 4,200, 4,300, 4,400, 4,500, 4,600, 4,700, 4,800, 4,900, 5,000, 5,100, 5,200, 5,300, 5,400, 5,500, 5,600, 5,700, 5,800, 5,900, 6,000, 6,100, 6,200, 6,300, 6,400, 6,500, 6,600, 6,700, 6,800, 6,900, or 7,000 liters or more. In some embodiments, the bioreactor vessel can include a volume of about 15 liters or less. In some embodiments, the bioreactor vessel can include a volume of about 50 liters or less. In some embodiments, the bioreactor vessel can include a volume of about 3,000 liters or more. In some embodiments, the bioreactor vessel can include a volume of about 6,000 liters or more.

[0290] In some embodiments, the total volume of the bioreactor vessel is greater than the working volume of the cell culture medium contained in the bioreactor. In some embodiments, the total volume of the bioreactor vessel can be about 10, 25, 50, 100, 500, 1,00, 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, 10,000, 11,000, 12,000, 13,000, 14,000, 15,000, 16,000, 17,000, 18,000, 19,000, 20,000 liters, or any range derivable therefrom or more.

[0291] In some embodiments, the bioreactor vessel is maintained at a set temperature. In some embodiments, the bioreactor vessel is maintained at about 20 to about 40 °C, about 30 to about 38 °C, or about 35 to about 38 °C. In certain embodiments, the bioreactor vessel is maintained at about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, or 43 °C. In certain embodiments, the bioreactor vessel is maintained at about 37 °C.

[0292] IV.B. Cell culture medium In some embodiments, systems and / or assemblies that may include a cell culture medium are described herein. In some embodiments, the cell culture medium can be any cell culture medium known in the art. In some embodiments, the cell culture medium can include cells, biologics, cell culture media, metabolites, and the like. In some embodiments, a feed stream of cell culture medium is added to the cell culture medium. In some embodiments, the cell culture medium can include certain amounts of concentrated cell nutrients such as sugars, peptides, vitamins, lipids, and the like. In some embodiments, fresh cell culture medium is introduced into the bioreactor shown in FIG. 2 via any input port 223. In some embodiments, the permeate cell culture medium contains lower concentrations of metabolites and / or higher concentrations of biologics than the cell culture medium and / or fresh cell culture medium. In some embodiments, the collected permeate cell culture medium contains higher concentrations of biologics than the perfusion permeate cell culture medium.

[0293] V. Methods of use Referring to FIG. 3, a non-limiting exemplary flowchart is shown that illustrates a method of producing a biologic that can operate during both the perfusion and collection processes using an HF filter. In certain embodiments, the HF filter is operable against tangential flow. In certain embodiments, the HF filter is a TFDF® filter. Method 300 can include, at step 302, growing a population of cells in a cell culture of a bioreactor that can produce a biologic and / or promote the production of a biologic (e.g., capable of growing a virus population, acting as a feeder cell, etc.). During step 304, the cells are grown using a perfusion process cell culture technique, and the cell culture fluid is passed through an HF filter to remove spent cell culture medium (e.g., the medium is perfused through the HF filter membrane), the cells are retained in the HF filter lumen, and the retained fluid stream is circulated back to the bioreactor. During the perfusion stage, the spent cell culture medium is perfused through the HF filter at a first flow rate to enter a first flow path (e.g., the flow path of the perfusion permeate). In certain embodiments, an appropriate replacement volume of cell culture medium is added to the bioreactor to maintain a desired cell culture fluid level in the bioreactor. In certain embodiments, the replacement volume of cell culture medium is approximately the same as, less than, or greater than the volume of spent cell culture medium removed. In certain embodiments, the replacement volume of cell culture medium is fresh cell culture medium. During step 306, following the growth of the cells using the perfusion process, a collection process is initiated. In certain embodiments, an intermediate process is performed between the perfusion process and the collection process. During such a collection process, the cell culture medium is perfused through the HF filter at a second flow rate to enter a second flow path (e.g., the flow path of the collection permeate). In certain embodiments, an appropriate replacement volume of cell culture medium is added to the bioreactor to maintain a desired cell culture fluid level in the bioreactor during different stages of collection. In certain embodiments, the replacement volume of cell culture medium is approximately the same as, less than, or greater than the volume of spent cell culture medium removed.In certain embodiments, the amount of cell culture medium exchanged is fresh cell culture medium. The second flow rate associated with the second flow path is greater than the first flow rate associated with the first flow path. In certain embodiments, the first flow rate of the first flow path is about 5 LPM or less, and the second flow rate of the second flow path is greater than about 5 LPM. In some embodiments, the method includes the use of any of the systems or assemblies described herein.

[0294] In some embodiments, the systems and devices described herein can be utilized in various processing modes including, but not limited to, perfusion, batch, fed-batch, semi-continuous processing (e.g., continuous harvesting, extended harvesting, etc.), and / or continuous processing.

[0295] In some embodiments, the systems and devices described herein are utilized in methods involving high liquid volumes, high liquid flow rates, high fluidity, and / or high pressures, and are suitable for use with sensitive cells (e.g., animal cells, such as mammalian cells) that tend to be sheared at high liquid volumes, high liquid flow rates, high fluidity, and / or high pressures.

[0296] V.A. Cells The assemblies and systems described herein can be used with any type of cell. In certain embodiments, the cells are considered to be relatively sensitive cells when compared to cells such as Escherichia coli or Saccharomyces cerevisiae (e.g., the cells are shear sensitive, e.g., the cells tend to shear at certain high volumes, flow rates, flow velocities, and / or pressures). In certain embodiments, the cells are animal cells. In certain embodiments, the cells are mammalian cells. In certain embodiments, the cells are immune effector cells (e.g., lymphocytes). In certain embodiments, the cells are T cells. In certain embodiments, the cells are B cells. In certain embodiments, the cells are NK cells. In certain embodiments, the cells are stem cells. In certain embodiments, the cells are induced pluripotent stem cells (iPSCs). In certain embodiments, the cells are stem cells and / or iPSCs that have differentiated into the hematopoietic lineage. In certain embodiments, the cells are hematopoietic progenitor cells. In certain embodiments, the cells are Chinese hamster ovary (CHO) cells. In certain embodiments, the cells are human embryonic kidney 293 (HEK293) cells. In certain embodiments, the cells are human fibrosarcoma cells (e.g., HT-1080 cells). In certain embodiments, the cells are derived from immortalized human embryonic cells (e.g., PER.C6 cells). In certain embodiments, the cells are a fusion of HEK293-S and a human B cell line (e.g., HKB-11 cells). In certain embodiments, the cells are derived from human hepatocellular carcinoma cells (e.g., HuH-7 cells). In certain embodiments, the cells are feeder cells and / or host cells for producing one or more viruses.

[0297] V.B. Products The assemblies and systems described herein can be used to produce products. In certain embodiments, the product is a cell (e.g., a stem cell, a cell of the hematopoietic lineage, an immune effector cell, etc.). In certain embodiments, the product is a biologic. In certain embodiments, the biologic can be of any type. In certain embodiments, the biologic is a small molecule, a peptide, a protein, an antibody (including, but not limited to, antibody derivatives such as Fc fusions, scFvs, multispecific antibodies, bispecific antibodies, etc.), a carbohydrate, a lipid, a virus, a virus-like particle, a viral protein / peptide, an extracellular particle (e.g., a microvesicle, an exosome, etc.), a vaccine, and / or a nucleotide (e.g., a DNA and / or an RNA molecule), but is not limited thereto.

[0298] In certain embodiments, the biologic comprises, consists essentially of, or consists of an antibody or a functional unit thereof (e.g., a single-chain variable fragment (scFv), a heavy chain, a light chain, a crystallizable fragment (Fc) domain, etc.). In certain embodiments, the biologic comprises, consists essentially of, or consists of a viral vector. In certain embodiments, the biologic comprises, consists essentially of, or consists of an adeno-associated virus (AAV). In certain embodiments, the biologic comprises, consists essentially of, or consists of a lentivirus. In certain embodiments, the biologic comprises, consists essentially of, or consists of an adenovirus. In certain embodiments, the biologic does not comprise, consist essentially of, or consist of AAV, lentivirus, and / or adenovirus. In certain embodiments, the biologic comprises, consists essentially of, or consists of an RNA molecule, e.g., but not limited to, a messenger RNA (mRNA), a small interfering RNA (siRNA), a microRNA (miRNA), a long non-coding RNA (lncRNA), and / or an antisense oligonucleotide (ASO). In some embodiments, the biologic comprises, consists essentially of, or consists of an antigenic molecule (e.g., a molecule capable of inducing an immune response in a subject).

[0299] V.C. Automation In some embodiments, the assemblies and systems described herein and methods of using them can be automated. In certain embodiments, the automation includes the integration of one or more human-machine interfaces (HMIs) with a processing device. In some embodiments, one or more sensors can provide real-time data to the processing device that can be displayed on the HMI. In some embodiments, the actions required to change the process of biomanufacturing, such as a perfusion process, an in-between process, or a harvesting process, can be programmed into the processing device and controlled via the HMI.

[0300] V.D. Timing In some embodiments, the assemblies and systems described herein and methods of using them can be of any duration. In certain embodiments, a perfusion process that includes the use of the multi-purpose assembly described herein can be about 0.25, 0.5, 1, 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, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days or more, or longer than 60 days, or any derivable range therein. In certain embodiments, the perfusion process is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 days, or more than 10 days, or any derivable range therein. In certain embodiments, the perfusion process is about 5 days. In certain embodiments, the perfusion process is about 6 days.

[0301] In certain embodiments, the HF filter is not exchanged between the perfusion process and the collection process. In certain embodiments, the collection process, including the use of the multi-purpose assembly described herein, can be about 0.25, 0.5, 1, 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, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days or more, or longer than 60 days, or any derivable range therein. In certain embodiments, the perfusion process and the collection process, including the use of the multi-purpose assembly described herein, can be about 0.25, 0.5, 1, 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, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, or 70 days or more, or longer than 70 days, or any derivable range therein. In certain embodiments, the collection process is about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, or longer than about 10 days. In some embodiments, the collection process is about 6 days. In some embodiments, the collection process is longer than about 6 days. In some embodiments, the collection process is a continuous collection process.

[0302] VI. Representative Experimental Results VI.A. Systems Containing a Multi-Purpose Assembly A multi-purpose assembly and system including the components shown in FIGS. 1 and 2 were produced. Biomanufacturing tests were conducted on at least 10 different biopharmaceuticals, including at least 4 tests involving a combined perfusion process and a continuous collection process. An exemplary multi-purpose assembly and system including an achieved filter flux level of up to 600 LMH, a VVD speed of up to 4.3, a VCD speed of up to 158×10 6 of, and a filter throughput speed of up to 57,500 L / m 2 . [Table 2]

[0303] VII. Further Considerations The headers and / or sub-headers between the sections and sub-sections of this document are included solely for the purpose of improving readability and do not imply that the features cannot be combined across sections and sub-sections. Thus, the sections and sub-sections do not describe separate embodiments.

[0304] Although the present teachings are described in relation to various embodiments, the present teachings are not intended to be limited to such embodiments. On the contrary, the present teachings include various changes, modifications, and equivalents, as will be apparent to those skilled in the art. This description is provided to offer a preferred exemplary embodiment and is not intended to limit the scope, applicability, or configuration of the present disclosure. Rather, the description of the preferred exemplary embodiment provides those skilled in the art with a possible explanation for implementing various embodiments.

[0305] It is understood that the functions and arrangements of elements may be variously changed without departing from the spirit and scope as described in the appended claims. Accordingly, such changes and modifications are considered to be within the scope described in the appended claims. Further, the terms and expressions used are used as terms of description and not of limitation, and there is no intention to exclude equivalents or portions thereof of the features shown and described in the use of such terms and expressions, but it is recognized that various changes are possible within the scope of the invention described in the claims.

[0306] In describing various embodiments, this specification may present a method and / or process as a particular series of steps. However, unless the method or process depends on the particular order of steps described herein, the method or process should not be limited to the particular order of steps described, and as would be readily apparent to one of ordinary skill in the art, the order may be changed and still remain within the spirit and scope of the various embodiments.

[0307] Some embodiments of the present disclosure include a system that includes one or more data processors. In some embodiments, the system includes a non-transitory computer-readable storage medium that includes instructions that, when executed by the one or more data processors, cause the one or more data processors to execute some or all of one or more of the methods and / or some or all of one or more of the processes disclosed herein. Some embodiments of the present disclosure include a computer program product tangibly embodied in a non-transitory machine-readable storage medium that includes instructions configured to cause one or more data processors to execute some or all of one or more of the methods and / or some or all of one or more of the processes disclosed herein.

[0308] To provide an understanding of the embodiments, specific details are given in this specification. However, it is understood that the embodiments may be practiced without these specific details. For example, circuits, systems, networks, processes, and other components may sometimes be shown as components in block diagram form so as not to obscure the embodiments with unnecessary detail. In other instances, well-known circuits, processes, algorithms, structures, and techniques may be shown without unnecessary detail in order to avoid obscuring the embodiments.

Claims

1. A bioreactor system capable of operating in a perfusion process and a collection process, a) Bioreactor including input and output ports, b) A supply conduit operably connected to the output port and inlet of a hollow fiber (HF) filter, the supply conduit operable to deliver fluid from the bioreactor to the HF filter, c) A retaining fluid conduit operably connected to the input port of the bioreactor and the outlet of the HF filter, the retaining fluid conduit operable to transport fluid from the HF filter to the bioreactor, and d) A multipurpose assembly operably connected to the HF filter, comprising a multipurpose assembly including a first flow path and a second flow path, The first flow path includes a first pump operable to draw fluid from the HF filter and a flow meter operable to measure the flow rate of fluid in the first flow path, the first flow path including the first pump draws perfusion fluid from the HF filter, The second flow path includes a second pump operable to draw fluid from the HF filter, and the second flow path including the second pump draws perfusion collection fluid from the HF filter. The first pump and the second pump are configured to have different pump capacities, and the second flow path is capable of guiding fluid at a higher flow rate than the first flow path. Bioreactor system.

2. The bioreactor system according to claim 1, wherein the HF filter is a tangential flow deep filtration (TFDF) filter.

3. The bioreactor system according to claim 1, further comprising one or more clarification filters for the first and / or second flow channels.

4. The bioreactor system according to claim 1, wherein the first and second pumps are selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetically driven pumps, positive displacement pumps, membrane pumps, pressure pumps, Quantex® pumps, gear pumps, diaphragm pumps, syringe pumps, and piston pumps.

5. The bioreactor system according to claim 1, wherein the first pump and the second pump are peristaltic pumps.

6. The bioreactor system according to claim 1, wherein the supply flow conduit comprises a supply flow pump selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetically driven pumps, positive displacement pumps, membrane pumps, pressure pumps, Quantex® pumps, gear pumps, diaphragm pumps, syringe pumps, and piston pumps, and is operable to deliver unfiltered fluid from the bioreactor to the HF filter.

7. The bioreactor system according to claim 1, wherein the retaining fluid flow conduit is operable to transport cell culture medium from the HF filter to the bioreactor.

8. The bioreactor system according to claim 1, wherein the first channel and the second channel have the same inner diameter or different inner diameters.

9. The first pump, from the HF filter, a. At a flow rate of approximately 0.01 LPM to approximately 15 LPM; b. At a rate of 0.1 to approximately 5 bioreactor container volumes / day (VVD); and / or c. With a filter flow rate of approximately 50 to 800 liters / square meter / hour (LMH) The bioreactor system according to claim 1, which is operable to draw out fluid.

10. The first pump is approximately 75 x 10 ６ The bioreactor system according to claim 1, which is capable of operating to draw fluid from the HF filter while maintaining a live cell density (VCD) exceeding cells / mL.

11. The bioreactor a. A volume of approximately 15 or approximately 50 liters or less; or b. A volume of approximately 500 liters or more A bioreactor system according to claim 1, comprising:

12. The bioreactor system according to claim 1, further comprising one or more of a supply flow or retaining fluid flow fluid pumps.

13. The bioreactor system according to claim 1, further comprising at least one component capable of facilitating at least one or more process enhancement parameters selected from an increase in cell number, an increase in cell density, a supply of abundant cell culture growth medium, a rapid expansion of cell numbers, or an increase in the production of a bioagent.

14. The bioreactor system according to claim 1, further comprising the ability to back-flushing the HF filter with a permeable fluid.

15. The second pump described above, a. Approximately 10,000 liters / square meter (L / m ２ ) ~ approx. 70,000L / m ２ With throughput equal to; b. With a filter flow rate of approximately 150 to approximately 900 liters / square meter / hour (LMH); and / or c. At a flow rate of approximately 0.01 LPM to approximately 18 LPM The bioreactor system according to claim 1, which is operable to draw fluid from the HF filter.

16. The bioreactor system according to claim 1, wherein the second flow path and pump are operable to draw a supply flow rate of about 1 to about 3 liters / fiber / min (L / fiber / min) from the HF filter.

17. The bioreactor system according to claim 1, for use with mammalian cells, in particular with Chinese ovarian hamster (CHO) cells and / or with human fetal kidney 293 (HEK293) cells.

18. The bioreactor system according to claim 1, which is operable for use with cells for the production of biological products, in particular antibodies, peptides, and / or viruses.

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