Systems, devices and methods for cell culture

Abstract

Embodiments described herein generally relate to components, systems, and methods for cell culture and production of biologics. Multi-purpose assemblies that facilitate a combination of a perfusion process and a harvest process are disclosed herein. The multi-purpose component and the system including the multi-purpose component simplify and streamline the bio-manufacturing process at least by improving automation capability, reducing physical footprint, reducing pollution risk, and reducing consumable usage. Methods of using the above assemblies and systems are also disclosed herein.

Description

Cross-references

[0001] This application is a PCT application claiming the benefit of 63 / 357,593, filed June 30, 2022, entitled “SYSTEMS, APPARATUS AND METHODS FOR CELL CULTURE” and is incorporated herein by reference in its entirety for all purposes. Technical Field

[0002] The present disclosure generally relates to apparatus, systems and methods for producing biologics. More specifically, the present disclosure relates to improving the use of cell retention devices in biomanufacturing apparatus, systems and methods. Background Art

[0003] Cell culture (e.g., culture of mammalian cells, bacterial cells, or fungal cells) can be performed to harvest living cells for therapeutic purposes and / or to harvest biomolecules, such as proteins or chemicals (e.g., biologics) produced by the cells. During a biomanufacturing process that includes cell culture, filtration is performed to separate, clarify, modify, and / or concentrate a fluid solution, mixture, or suspension. In the biotechnology and pharmaceutical industries, filtration is essential for the successful production, processing, and testing of new drugs, diagnostic reagents, and other biologics. For example, in the process of using animal or microbial cell culture to manufacture biologics, filtration is performed for clarification, selective removal, and / or concentration of certain components from the culture medium, and / or to modify the culture medium before further processing. Filtration can also be used to increase productivity by maintaining the culture at a high cell concentration during perfusion.

[0004] One use of a filtration unit may be as a cell retention device (CRD). Well-known CRDs include tangential flow filtration (also known as cross-flow filtration or TFF) systems. Such systems are widely used to separate particulates suspended in a liquid phase and have important biomanufacturing applications. In contrast to dead-end filtration systems, in which a single fluid feed is passed through the filter, a tangential flow system is characterized by the flow of the fluid feed across the surface of the filter, resulting in the separation of the feed into two components: a permeate component that has passed through the filter and a retentate component that has not passed through the filter. TFF systems are less prone to fouling than dead-end systems. Fouling of TFF systems can be further reduced by alternating the direction of the fluid feed through the filter assembly, as is done in the alternating tangential flow (ATF) technique, by backwashing the permeate passing through the filter, and / or by periodically washing the filter.

[0005] Advances in filtration have been made (see, e.g., Tim Sewerin et al., "Advances and Applications of Hollow Fiber Nanofiltration Membranes: A Review. Membranes, 2021, 11, 890). One such advancement includes hollow fiber (HF) filters (HFFs) that can be operated in a tangential flow mode, one example of which is a Tangential Flow Depth Filtration® (TFDF®) filter. Such TFDF® filters combine the advantages of tangential flow and depth filtration. However, some HF filters may be considered expensive and / or relatively bulky, and cell cultures are susceptible to contamination and associated product losses during the transition between bioreactors used for cell growth phases and harvest phases.

[0006] Therefore, it is desirable to have components, systems, and methods that can improve these processes. Summary of the invention

[0007] Components, systems and methods suitable for producing biologics are described herein. Bioreactor systems are disclosed herein, comprising: a) a bioreactor comprising an input port and an output port; b) a feed flow conduit operably connected to the output port and an inlet of a hollow fiber (HF) filter, the feed flow conduit being operable to transport fluid from the bioreactor to the HF filter; c) a retentate flow conduit operably connected to the input port of the bioreactor and an outlet of the HF filter, the retentate flow conduit being operable to transport 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: wherein the first flow path comprises a first pump operable to extract fluid from the HF filter and a flow meter operable to measure the flow rate of fluid in the first flow path; wherein the second flow path comprises a second pump operable to extract fluid from the HF filter; and wherein the first pump and the second pump are configured to have different pump capacities.

[0008] In some embodiments, the HF filter is operable for tangential flow. In some embodiments, the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter. In some embodiments, the first pump and the second pump are configured to have different precision levels. In some embodiments, the system further comprises: one or more clarifying filters in the first flow path and / or the second flow path. In some embodiments, the first pump and the second pump are selected from the group consisting of: a peristaltic pump, a centrifugal pump, a magnetically driven pump, a positive displacement pump, a membrane pump, a pressure-based pump, a quantex™ pump (e.g., a positive displacement rotary pump), a gear pump, a diaphragm pump, a syringe pump, and a piston pump. In some embodiments, the first pump and the second pump are peristaltic pumps. In some embodiments, the feed flow conduit comprises a feed flow pump selected from the group consisting of: a peristaltic pump, a centrifugal pump, a magnetically driven pump, a positive displacement pump, a membrane pump, a pressure-based pump, a quantex™ pump, a gear pump, a diaphragm pump, a syringe pump, and a piston pump, the feed flow conduit being operable to convey unfiltered fluid from the bioreactor to the HF filter. In some embodiments, the retentate stream conduit is operable to transport cell culture fluid from the HF filter to the bioreactor.

[0009] In some embodiments, a first flow path comprising a first pump operable to extract fluid from the HF filter extracts a perfusion permeate flow fluid from the HF filter. In some embodiments, a second flow path comprising a second pump operable to extract fluid from the HF filter extracts a harvest 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 is capable of conducting 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 between about 0.01 liters per minute (LPM) and 5 LPM, between about 0.01 LPM and about 10 LPM, and / or between about 0.01 LPM and about 15 LPM. In some embodiments, the first flow path and pump are operable to draw a feed rate from the HF filter of about 0.8 liters per minute per fiber to about 2.2 liters per minute per fiber (L / fiber / min). In some embodiments, the first pump is operable to draw fluid from the HF filter while maintaining a flow rate of greater than about 75 x 10 6 cells / mL, greater than about 50 x 10 6 cells / mL and / or greater than approximately 25 x 10 6In some embodiments, the first pump is operable to draw fluid from the HF filter at a rate between about 0.1 and about 5 bioreactor vessel volumes per day (VVD), at a rate between about 0.5 and about 4.3 bioreactor VVD, at a rate between about 1 and about 4.3 bioreactor VVD, and / or at a rate of up to about 5 bioreactor VVD. In some embodiments, the first pump is operable to draw fluid from the HF filter at a filter flux rate between about 50 liters per square meter per hour and about 800 liters per square meter per hour (LMH) and / or between about 100 LMH and 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 rate to maintain a desired VVD rate. In some embodiments, the flow meter operable to measure the flow rate of the fluid in the first flow path is capable of accurately monitoring a flow rate of about 0 LPM to about 8 LPM or about 0.5 LPM to about 6 LPM. In some embodiments, the first pump is operable to pump at a rate equal to about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 Between 10,000 liters per square meter (L / m 2 ) to about 50,000 L / m 2 Between and / or around 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 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 less than or equal to about 15 liters, less than or equal to about 50 liters, less than or equal to about 100 liters, or less than or equal to about 500 liters. In some embodiments, the bioreactor has a volume of greater than or equal to about 500 liters, about 1,000 liters, about 1,500 liters, and / or about 3,500 liters. In some embodiments, the bioreactor has a volume equal to between about 2,000 liters and 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 be operated during a perfusion process. In some embodiments, the bioreactor system can be operated during a harvest process. In some embodiments, the bioreactor system can be operated during a perfusion process, an intermediate perfusion and harvest process, and a harvest process and a harvest process. In some embodiments, the bioreactor system can be operated during a continuous harvest process. In some embodiments, the bioreactor system further includes a human-machine interface (HMI) control unit. In some embodiments, the HMI control unit is programmed and can display a perfusion process control, an intermediate perfusion and harvest process control, or a harvest process control.

[0014] In some embodiments, the bioreactor system further comprises one or more sensors. In some embodiments, the bioreactor system further comprises one or more of a feed stream fluid pump or a retentate stream fluid pump. In some embodiments, the bioreactor system further comprises at least one component capable of promoting at least one process intensification parameter. In some embodiments, the process intensification parameter is one or more of an increased cell number, an increased cell density, a provision of an enriched cell culture growth medium, a rapid cell number expansion, or an increased biopharmaceutical yield. In some embodiments, the bioreactor system is further capable of backwashing the HF filter with a 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 the flow rate from about 0 LPM to about 10 LPM or from about 0.5 LPM to about 8 LPM. In some embodiments, the second pump is operable to pump at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 Between 12,000 liters per square meter (L / m 2 ) to about 60,000 L / m 2and / or at approximately 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 throughput rate between about 150 liters per square meter per hour and about 900 liters per square meter per hour (LMH) and / or between about 200 LMH and about 700 LMH.

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

[0017] In some embodiments, the first flow path includes a connection system having 3 / 8" ID tubing and / or 1 / 8" ID tubing. In some embodiments, the second flow path includes a connection system having 1 / 2" ID tubing and / or 1 / 8" ID tubing. In some embodiments, the first flow path has an accuracy requirement of approximately 1%. In some embodiments, the second flow path has an accuracy requirement of approximately 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 between about 0.65 μm and 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.

[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 fetal kidney 293 (HEK293) cells. In some embodiments, the bioreactor system is operable for use with cells to produce biologics. In some embodiments, biologics include antibodies, peptides, and / or viruses.

[0020] Also disclosed herein are multi-purpose filter assemblies comprising: a) a hollow fiber (HF) filter; and b) a multi-purpose assembly operably connected to the HF filter, the multi-purpose assembly comprising a first flow path and a second flow path: wherein the first flow path comprises a first pump operable to extract fluid from the HF filter and a flow meter operable to measure the flow of fluid in the first flow path; wherein the second flow path comprises a second pump operable to extract fluid from the HF filter; and wherein the first pump and the second pump can have different pump capacities and accuracy levels. In some embodiments, the HF filter is operable for tangential flow.

[0021] In some embodiments, the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter. In some embodiments, the multi-purpose filter assembly further comprises one or more clarifying filters in the first flow path and / or the second flow path. In some embodiments, the first pump and the second pump 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-based pump, a quantex™ pump, a gear pump, a diaphragm pump, a syringe pump, and a piston pump. In some embodiments, the first pump and the second pump are peristaltic pumps. In some embodiments, the first flow path and the pump are operable to pump at a flow rate greater than or equal to about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 In some embodiments, the first flow path and the pump are operable to draw fluid from the HF filter at a throughput of greater than or equal to about 10,000 liters per square meter (L / m 2 ) to about 50,000 L / m 2 In some embodiments, the first flow path and the pump are operable to draw fluid from the HF filter at a throughput of greater than or equal to about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2In some embodiments, the first flow path and pump are operable to draw fluid from the HF filter at a filter throughput of about 50 to about 800 LMH and / or about 100 to about 600 LMH. In some embodiments, the first flow path and pump are operable to maintain a hematocrit (PCV) of about 2% to about 40%, about 8% to about 40%, and / or about 12% to about 35%. In some embodiments, the first flow path and pump are operable to draw fluid from the HF filter while maintaining a PCV greater than about 25 x 10 6 cells / mL, approximately 50 x 10 6 cells / mL, approximately 75 x 10 6 cells / mL, approximately 100 x 10 6 cells / mL and / or approximately 25 x 10 7 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 between about 0.01 LPM to about 5 LPM, about 0.01 LPM to about 10 LPM, and / or about 0.01 LPM to about 15 LPM.

[0022] In some embodiments, a flow meter operable to measure the flow of fluid in the first flow path continuously monitors the LPM rate and communicates with the first pump to adjust the first pump rate to maintain a desired LPM rate. In some embodiments, the flow meter operable to measure the flow of fluid in the first flow path is capable of accurately monitoring a flow rate of about 0 LPM to about 8 LPM and / or 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 the pump are operable to extract fluid from the HF filter with a higher flow rate than the first flow path and the first pump. In some embodiments, the second flow path and the pump are operable to extract fluid from the HF filter as a continuous harvesting process. In some embodiments, when the multi-purpose assembly guides fluid through a flow path and pump and then guides fluid through another flow path and pump, the HF filter is not replaced. In some embodiments, when the multi-purpose assembly guides fluid first through the first flow path and pump and then through the second flow path and pump, the HF filter is not replaced. In some embodiments, 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.

[0024] In some embodiments, the multi-use 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 the flow rate from about 0 LPM to about 10 LPM or from about 0.5 LPM to about 8 LPM. In some embodiments, the second pump is operable to pump at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 Between 12,000 L / m 2 to about 60,000 L / m 2 Between and / or around 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 between about 150 liters per square meter per hour and about 900 liters per square meter per hour (LMH) and / or between about 200 LMH and about 700 LMH. In some embodiments, the second flow path and pump are operable to draw a feed flow rate from the HF filter of about 1 liter per fiber per minute to about 3 liters per fiber per minute (L / fiber / min). In some embodiments, the second pump is operable to draw fluid from the HF filter at a flow rate between 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 between about 0.65 μm and 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 to produce biologics. In some embodiments, the biologics include antibodies, peptides, and / or viruses.

[0027] Also disclosed herein are methods for producing biologics from cells, comprising: a) expanding cells in a cell culture fluid in a bioreactor, b) during cell expansion, perfusing the cell culture fluid through a hollow fiber (HF) filter to remove spent cell culture medium while retaining expanding cells, and adding an appropriate replacement volume of cell culture medium to the bioreactor to maintain a desired cell culture fluid content in the bioreactor, wherein the spent cell culture medium is perfused through the HF filter at a first flow rate to enter a first flow path, and c) after cell expansion, harvesting 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 content in the bioreactor during different harvest stages, wherein the cell culture medium is harvested through the HF filter at a second flow rate to enter a second flow path, wherein the second flow rate is greater than the first flow rate. In some embodiments, the HF filter is operable for tangential flow. In some embodiments, the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter. In some embodiments, the replacement volume of the cell culture medium in b) and / or c) is the same as the amount of spent cell culture medium removed. In some embodiments, the replacement volume of the 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 the 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 the cell culture medium is fresh cell culture medium. In some embodiments, the first flow rate is less than or equal to about 5 LPM and the second flow rate is greater than about 5 LPM. In some embodiments, the HF filter is made 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 between about 0.65 μm and 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 cell is an animal cell, a mammalian cell, a CHO cell and / or a HEK293 cell. In some embodiments, the biologic comprises an antibody, a peptide and / or a virus.

[0028] Also disclosed herein are methods of producing biologics from cells comprising using the bioreactor systems and / or multi-use filter assemblies 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 a bioreactor system of the present disclosure or 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 and the use may further comprise harvesting the cell culture fluid through a HF filter to obtain the biologic.

[0031] Also disclosed herein are methods for producing biologics from cells, which methods include using the bioreactor system or multi-purpose filter assembly disclosed herein. Also disclosed herein are methods for using the systems, assemblies, or methods disclosed herein to increase productivity, improve performance, increase efficiency, reduce contamination rates, reduce capital investment, reduce physical footprint, reduce consumables loss, and / or reduce operating costs for facilities including biologics production processes.

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

[0033] Aspect 1 is a bioreactor system, which includes: a) a bioreactor, which includes an input port and an output port; b) a feed flow conduit, which is operably connected to the output port and the inlet of a hollow fiber (HF) filter, the feed flow conduit being operable to transport fluid from the bioreactor to the HF filter; c) a retentate flow conduit, which is operably connected to the input port of the bioreactor and the outlet of the HF filter, the retentate flow conduit being operable to transport fluid from the HF filter to the bioreactor; and d) a multi-purpose component, which is operably connected to the HF filter, the multi-purpose component including a first flow path and a second flow path: wherein the first flow path includes a first pump operable to extract fluid from the HF filter and a flow meter operable to measure the flow rate of the fluid in the first flow path; wherein the second flow path includes a second pump operable to extract fluid from the HF filter; and wherein the first pump and the second pump are configured to have different pump capacities.

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

[0035] Aspect 3 is a bioreactor system according to aspect 1 or 2, wherein the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter.

[0036] Aspect 4 is a bioreactor system according to any one of aspects 1 to 3, wherein the first pump and the second pump are configured to have different accuracy levels.

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

[0038] Aspect 6 is a bioreactor system according to any one of aspects 1 to 5, wherein the first pump and the second pump 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-based pump, a quantex™ pump, a gear pump, a diaphragm pump, a syringe pump, and a piston pump.

[0039] Aspect 7 is a 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 a bioreactor system according to any one of aspects 1 to 7, wherein the feed flow conduit includes a feed 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-based pump, a quantex™ pump, a gear pump, a diaphragm pump, a syringe pump and a piston pump, and the feed flow conduit is operable to transport unfiltered fluid from the bioreactor to the HF filter.

[0041] Aspect 9 is a bioreactor system according to any one of aspects 1 to 8, wherein the retentate stream conduit is operable to transport the cell culture fluid from the HF filter to the bioreactor.

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

[0043] Aspect 11 is a bioreactor system according to any one of aspects 1 to 10, wherein a second flow path comprising a second pump operable to extract fluid from the HF filter extracts a harvest permeate stream fluid from the HF filter.

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

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

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

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

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

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

[0050] Aspect 18 is a 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 flow rate greater than about 75 x 10 6 The viable cell density (VCD) was calculated as cells / mL.

[0051] Aspect 19 is a 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 flow rate greater than about 50 x 10 6 The viable cell density (VCD) was calculated as cells / mL.

[0052] Aspect 20 is a 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 flow rate greater than about 25 x 10 6 cells / mL of VCD.

[0053] Aspect 21 is a 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 between about 0.1 and about 5 bioreactor vessel volumes per day (VVD).

[0054] Aspect 22 is a 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 between about 0.5 and about 4.3 bioreactor vessel volumes per day (VVD).

[0055] Aspect 23 is a 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 flux rate between about 50 liters per square meter per hour and about 800 liters per square meter per hour (LMH).

[0056] Aspect 24 is a 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 flux rate of between about 100 LMH and about 600 LMH.

[0057] Aspect 25 is a bioreactor system according to any one of aspects 1 to 24, wherein a flow meter operable to measure the flow rate of 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 a desired VVD rate.

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

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

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

[0061] Aspect 29 is a 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 a 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 a 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 a bioreactor system according to any one of aspects 1 to 31, wherein the bioreactor has a volume of less than or equal to about 15 liters, less than or equal to about 50 liters, less than or equal to about 100 liters, or less than or equal to about 500 liters.

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

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

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

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

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

[0070] Aspect 38 is a 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 a bioreactor system according to any one of aspects 1 to 38, wherein the bioreactor system is capable of operating in a perfusion process.

[0072] Aspect 40 is a bioreactor system according to any one of aspects 1 to 39, wherein the bioreactor system is capable of operating during a harvesting process.

[0073] Aspect 41 is a bioreactor system according to any one of aspects 1 to 40, wherein the bioreactor system is capable of operating during a perfusion process, an intermediate perfusion and harvest process, and a harvest process.

[0074] Aspect 41.1 is a bioreactor system according to any one of aspects 1 to 41, wherein the bioreactor system is capable of operating in more than one perfusion process.

[0075] Aspect 41.2 is a bioreactor system according to any one of aspects 1 to 41.1, wherein the bioreactor system is capable of operating in more than one perfusion process, intermediate perfusion and harvest process, and harvest process.

[0076] Aspect 42 is a bioreactor system according to any one of aspects 1 to 41.2, wherein the bioreactor system is capable of operating in a continuous harvesting process.

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

[0078] Aspect 44 is a bioreactor system according to aspect 43, wherein the HMI control unit is programmed and can display perfusion process control, intermediate perfusion and harvest process or harvest process control.

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

[0080] Aspect 46 is a bioreactor system according to any one of aspects 1 to 46, further comprising one or more of a feed stream fluid pump or a retentate stream fluid pump.

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

[0082] Aspect 48 is a bioreactor system according to aspect 47, wherein the process intensification parameter is one or more of increased cell number, increased cell density, provision of enriched cell culture growth medium, rapid cell number expansion, or increased biologic yield.

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

[0084] Aspect 50 is a 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 LPM to about 10 LPM or about 0.5 LPM to about 8 LPM.

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

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

[0087] Aspect 53 is a 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 flux rate (LMH) between about 150 liters per square meter per hour and about 900 liters per square meter per hour.

[0088] Aspect 54 is a 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 flux rate of between about 200 and about 700 LMH.

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

[0090] Aspect 56 is a 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 between about 0.01 LPM to about 8 LPM.

[0091] Aspect 57 is a 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 between about 0.01 LPM to about 13 LPM.

[0092] Aspect 58 is a 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 between about 0.01 LPM to about 18 LPM.

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

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

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

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

[0097] Aspect 63 is a 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 a 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 a bioreactor system according to any one of aspects 1 to 64, wherein the HF filter is constructed of and / or comprises polypropylene and / or polyethylene terephthalate.

[0100] Aspect 66 is a bioreactor system according to any one of aspects 1 to 65, wherein the HF filter comprises an isotropic pore structure and / or comprises 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 a bioreactor system according to any one of aspects 1 to 66, operable for use with shear sensitive cells.

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

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

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

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

[0106] Aspect 72 is a bioreactor system according to any one of aspects 1 to 71, operable for use with cells to produce a biologic.

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

[0108] Aspect 74 is a multi-purpose filter assembly, comprising: a) a hollow fiber (HF) filter; and b) a multi-purpose assembly operably connected to the HF filter, the multi-purpose assembly comprising a first flow path and a second flow path: wherein the first flow path comprises a first pump operable to extract fluid from the HF filter and a flow meter operable to measure the flow rate of the fluid in the first flow path; wherein the second flow path comprises a second pump operable to extract fluid from the HF filter; and wherein the first pump and the second pump are capable of having different pump capacities and accuracy levels.

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

[0110] Aspect 76 is a multi-use filter assembly according to aspect 74 or 75, wherein the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter.

[0111] Aspect 77 is a multi-use filter assembly according to any one of aspects 74 to 76, further comprising: one or more clarifying filters in the first flow path and / or the second flow path.

[0112] Aspect 78 is a multi-use filter assembly according to any one of Aspects 74 to 77, wherein the first pump and the second pump are selected from the group consisting of: peristaltic pumps, centrifugal pumps, magnetic drive pumps, positive displacement pumps, membrane pumps, pressure-based pumps, quantex™ pumps, gear pumps, diaphragm pumps, syringe pumps and piston pumps.

[0113] Aspect 79 is the multi-use 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 multi-use filter assembly of any one of aspects 74 to 79, wherein the first flow path and the pump are operable to pump the filter at a flow rate greater than or equal to about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 The fluid is drawn from the HF filter at a throughput of .

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

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

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

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

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

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

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

[0122] Aspect 88 is the multi-use filter assembly of 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-use filter assembly of 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 a multi-use filter assembly according to any one of aspects 74 to 89, wherein the first flow path and the pump are operable to extract fluid from the HF filter while maintaining a flow rate greater than about 25 x 10 6 cells / mL of VCD.

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

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

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

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

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

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

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

[0132] Aspect 98 is a multi-use 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 between about 0.01 LPM to about 5 LPM.

[0133] Aspect 99 is the multi-use filter assembly of 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 between about 0.01 LPM to about 10 LPM.

[0134] Aspect 100 is a multi-use 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 between about 0.01 LPM to about 15 LPM.

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

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

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

[0138] Aspect 104 is a multi-use 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 a multi-use 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 a multi-use filter assembly according to any one of aspects 74 to 105, wherein the first flow path and the second flow path have the same or different inner diameters.

[0141] Aspect 107 is a multi-use filter assembly according to any one of aspects 74 to 106, wherein 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 first pump.

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

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

[0144] Aspect 110 is the multi-use filter assembly according to aspect 109, wherein the HF filter is not replaced when the multi-use assembly directs fluid first through the first flow path and the pump and then through the second flow path and the pump.

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

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

[0147] Aspect 113 is a multi-use filter assembly according to any one of aspects 74 to 112, wherein the second pump is operable to pump the filter at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 The flow rate between draws fluid from the HF filter.

[0148] Aspect 114 is a multi-use filter assembly according to any one of aspects 74 to 112, wherein the second pump is operable to pump the filter at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 60,000 L / m 2 Between and / or around 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 The flow rate between draws fluid from the HF filter.

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

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

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

[0152] Aspect 118 is the multi-use 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 between about 0.01 LPM to about 8 LPM.

[0153] Aspect 119 is the multi-use 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 between about 0.01 LPM to about 13 LPM.

[0154] Aspect 120 is the multi-use 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 between about 0.01 LPM to about 18 LPM.

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

[0156] Aspect 122 is a multi-use filter assembly according to any one of aspects 74 to 121, wherein the HF filter comprises an isotropic pore structure and / or comprises an average pore cavity diameter of about 0.65 µm to about 8 µm or about 2 µm to about 5 µm.

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

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

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

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

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

[0162] Aspect 128 is a multi-use filter assembly according to any one of aspects 74 to 127, which is operable for use with cells to produce a biologic.

[0163] Aspect 129 is the multi-use filter assembly according to aspect 128, wherein the biological agent comprises an antibody, a peptide and / or a virus.

[0164] Aspect 130 is a method for producing a biologic from cells, the method comprising: a) expanding cells in a cell culture fluid in a bioreactor, b) during cell expansion, perfusing the cell culture fluid through a hollow fiber (HF) filter to remove spent cell culture medium while retaining expanding cells, and adding an appropriate replacement volume of cell culture medium to the bioreactor to maintain a desired cell culture fluid content in the bioreactor, wherein the spent cell culture medium is perfused through the HF filter at a first flow rate to enter a first flow path, and c) after cell expansion, harvesting 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 content in the bioreactor during different harvest stages, wherein the cell culture medium is harvested through the HF filter at a second flow rate to enter a second flow path, wherein the second flow rate is greater than the first flow rate.

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

[0166] Aspect 132 is a 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 a method according to any one of aspects 130 to 132, wherein the replacement volume of the cell culture medium in b) and / or c) is about the same as the amount of spent cell culture medium removed.

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

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

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

[0171] Aspect 136.1 is a method according to any one of aspects 130 to 136, wherein the perfusion process comprises more than one perfusion phase.

[0172] Aspect 136.2 is a method according to any one of aspects 130 to 136.1, wherein the perfusion process comprises more than one perfusion stage, and the HF filter is used in more than one perfusion stage, an intermediate perfusion and harvesting process, and a harvesting process.

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

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

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

[0176] Aspect 140 is a method according to any one of aspects 130 to 139, wherein the cell is a shear-sensitive cell.

[0177] Aspect 141 is a method according to any one of aspects 130 to 140, wherein the cell is an animal cell.

[0178] Aspect 142 is a method according to any one of aspects 130 to 141, wherein the cell is a mammalian cell.

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

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

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

[0182] Aspect 146 is a method of producing a biologic from cells, the method comprising using the bioreactor system or multi-use filter assembly according to any one of aspects 1 to 129.

[0183] Aspect 147 is use of the multi-use 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 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 cells produce a biologic and the use further comprises harvesting the cell culture fluid through a HF filter to obtain the biologic.

[0187] Aspect 151 is a system, component, method or use according to any one of aspects 1 to 150, wherein the system, component, method or use results in one or more of increased productivity, improved performance, increased efficiency, reduced contamination rate, reduced capital investment, reduced physical footprint, reduced consumables loss and / or reduced operating costs for a facility comprising a biologics production process. BRIEF DESCRIPTION OF THE DRAWINGS

[0188] The present disclosure is described in conjunction with the accompanying drawings:

[0189] FIG. 1 is a schematic diagram of an exemplary multi-purpose module apparatus.

[0190] FIG. 2 is a schematic diagram of an exemplary system including a multi-purpose component device.

[0191] FIG. 3 is a flow chart of an exemplary method described herein. DETAILED DESCRIPTION

[0192] I. Overview

[0193] The embodiments described herein recognize that biomanufacturing is an important component of biologics production, and therefore, an important component of modern medicine.

[0194] Cell retention devices (CRDs) play a key role in establishing perfusion processes. Typically, a CRD is able to retain cells in a bioreactor during the perfusion process while continuously supplying fresh media to the bioreactor and continuously withdrawing permeate (e.g., spent media) at a predetermined rate. Currently commercially available cell retention devices (e.g., ATF and TFF) are unable to successfully operate large cell (e.g., CHO cell) culture working volumes (e.g., ≥3000 L) with high perfusion rates (e.g., ≥2 vessel volumes per day (VVD)). These cell retention devices have a throughput of approximately 13 L / m 2 / h and a throughput of approximately 17,500 L / m 2 restrictions.

[0195] As described herein, in some embodiments, the inventors have shown that HF ​​filters (e.g., TFDF® filters) can be used as cell retention devices for large-scale cell (e.g., CHO cell) culture working volumes (e.g., ≥3000 L) with high perfusion rates (e.g., ≥2 vessel volumes per day (VVD)), resulting in higher cell densities and titers. As shown herein, the inventors have successfully created HF filter slides (e.g., TFDF® filter slides) with perfusion capabilities and tested them with parameters up to 600 L / m 2 / h throughput and 70,000 L / m 2HF filter (e.g., TFDF® filter) with a high volume of perfusion. In some embodiments, the inventors have found that systems including TFDF® are suitable for operation with up to 5 bioreactors VVD. In addition, the inventors have found that in some embodiments, the same HF filter (e.g., TFDF® filter) used during the perfusion process can also be used for a harvesting process, such as, but not limited to, a continuous harvest (CH) process (e.g., also referred to as an extended harvest process, etc.). To the best of the inventors' knowledge, this is the first time that such a process has been demonstrated to be accomplished at such high pressures and / or volumes in a process mode other than a harvesting mode. In some embodiments, the HF filter can be used for perfusion, batch, fed-batch, continuous and / or semi-continuous harvesting. In some methods including CH, before the end of a production run, permeate from the HF filter is collected for downstream processing while fresh cell culture medium (e.g., including metabolites / buffers) is perfused through the bioreactor, and at the end of a production run, the addition of fresh cell culture medium (e.g., including metabolites / buffers) is stopped and / or reduced while permeate extraction is still active, resulting in a reduction in bioreactor volume. In some embodiments, all of the cell culture fluid and / or all of the permeate is processed in a downstream process (DSP).

[0196] In some embodiments, systems, devices, and methods as described herein utilize HF filters to achieve higher working volumes (e.g., >3,000 L), higher VVD, increased media exchange rates during perfusion, higher throughput, multi-stage use (e.g., one or more perfusion stages and one or more harvest stages), and / or higher throughput relative to conventional systems, methods, and / or devices, while maintaining low cell shear rates and / or cell stress levels, thereby facilitating the cultivation of relatively sensitive cells (e.g., animal cells, e.g., cells without cell walls) that do not pass through the associated filters.

[0197] In some embodiments, described herein are techniques operable at high flow rates, large volumes, bioreactors of any size, and at any stage of the cell culture cycle, such as multi-purpose assemblies and / or systems. As described herein, in some embodiments, features associated with hollow fiber filters facilitate such capabilities (e.g., adaptability to high flow rates, large volumes, bioreactors of any size, and / or use at any stage of the cell culture cycle), and in certain embodiments, the hollow fiber filters are operable in tangential flow mode.

[0198] In some embodiments, described herein are technologies, such as multi-use components, that can be used to reduce contamination risk, reduce consumable wear rates, reduce the physical footprint associated with biomanufacturing, enable continuous or semi-continuous bioprocessing for purification, and / or simplify the biologics production process.

[0199] In certain embodiments, provided herein are techniques for reducing the physical footprint associated with biomanufacturing (e.g., the required manufacturing area), e.g., reducing the necessary footprint by equal to or greater than about 50%. In some embodiments, provided herein are methods, systems, and apparatus that can reduce the physical footprint associated with biomanufacturing by equal to or greater than about 66%. In some embodiments, the reduction in physical footprint is achieved by combining one or more biomanufacturing processes (e.g., perfusion processes and / or harvesting processes) into the same physical container and / or system using the systems and / or components described herein, e.g., using the same bioreactor used for the perfusion process for the harvesting process (e.g., batch, fed-batch, continuous, and / or semi-continuous harvesting).

[0200] In some embodiments, the technology described herein provides additional automation capabilities. For example, in some embodiments, the technology described herein avoids the physical transfer of cell culture fluid from one bioreactor to another bioreactor, and thus reduces the number of manual processes and increases the possibility of process automation.

[0201] In some embodiments, the techniques described herein provide improved efficiency. For example, in some embodiments, automated processes (e.g., cell density-triggered phase changes (e.g., inoculation, filling a bioreactor, adding an inducer, etc.)) can provide manufacturers with virtually no cell waste, process waste, and / or time waste. In some embodiments, the techniques described herein provide a reduction in labor / handling work. For example, in some embodiments, there is no need to prepare a dedicated harvesting device to receive the cell culture fluid (e.g., culture fluid) produced during the preparatory perfusion process because the harvesting system and the perfusion system are integrated.

[0202] In some embodiments, the technology described herein provides a reduction in the consumption rate of consumables. For example, in some embodiments, a reduction of necessary consumables equal to or greater than about 50% can be achieved. In some embodiments, the technology described herein provides a reduction in consumable costs, for example, because no additional cell retention device (CRD) is required for harvesting (e.g., using the same CRD during the perfusion phase and the harvesting phase), one CRD can be used for two processes. In some embodiments, the technology provided herein may include using the same CRD during a perfusion process, and the perfusion process includes more than one perfusion phase, such as but not limited to 1, 2, 3, 4, 5, 6 or more than 6 perfusion phases. In some embodiments, the perfusion process includes a series of perfusion phases (e.g., seed tank culture). In some embodiments, a series of perfusion phases may include an incremental increase in the bioreactor volume and / or viable cell density in each subsequent phase. In some embodiments, the technology provided herein can include using the same CRD for greater than or equal to 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 days or more. Thus, in some embodiments, the techniques described herein provide less consumable storage management effort and / or less storage space.

[0203] In some embodiments, the techniques described herein provide less risk of contamination. For example, in some embodiments, fewer manual applications and reduced transfer of cell culture fluids or biologic production materials reduce the likelihood of introducing contaminants.

[0204] In some embodiments, the techniques described herein provide a reduction in the number of CRDs required during a biomanufacturing process, particularly under conditions of large-scale biomanufacturing. For example, in some embodiments, a CRD used during a perfusion process can be used during a harvesting process (e.g., batch, fed-batch, continuous and / or semi-continuous harvesting). In some embodiments, a CRD (e.g., a HF filter) is already attached during a perfusion process and is reused during a harvesting phase (e.g., perfusion, batch, fed-batch, continuous harvesting, semi-continuous harvesting, bulk harvesting, etc.). In some embodiments, the CRD remains attached to the bioreactor during transitions between processes. In some embodiments, the CRD is in a "standby" mode, e.g., cells are still circulating through the lumen of the filter fibers, but no permeate is being drawn through the CRD.

[0205] The following description provides exemplary implementations of the devices, systems and methods described herein for biomanufacturing. Descriptions and examples of various terms as used herein are provided in Section II below.

[0206] II. Exemplary Description of Terms

[0207] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0208] Unless otherwise defined, all technical terms, symbols and other technical and scientific terms or expressions used herein are intended to have the meanings commonly understood by those of ordinary skill in the art to which the claimed subject matter belongs. In some cases, for the sake of clarity and / or for ease of reference, terms with commonly understood meanings are defined herein, and these definitions contained herein are not necessarily interpreted as representing significant differences from the common understanding of the art compared to the definitions of terms commonly understood in the prior art. In general, nomenclature and techniques used in conjunction with chemistry, biochemistry, molecular biology, pharmacology and toxicology are described herein, which are those well known and commonly used in the art.

[0209] Throughout the present disclosure, various aspects are presented in a range format. It should be understood that the description in a range format is only for convenience and brevity, and should not be interpreted as an inflexible limitation to the present disclosure. Therefore, the description of the range should be considered to have specifically disclosed all possible sub-ranges and each numerical value in the range. For example, in the case of providing a value range, it should be understood that each intermediate value between the upper and lower limits of the range and any other described value or intermediate value in the range is included in the present disclosure. The upper and lower limits of these smaller ranges can be independently included in the smaller range, and are also included in the present disclosure, but any explicit exclusion restrictions in the range will be observed. If the range includes one or two limits, then the scope excluding any one or two of the included restrictions is also included in the present disclosure. Regardless of the width of the range, this applies.

[0210] The headings and subheadings between sections and subsections of this document are used only for the purpose of improving readability and do not imply that features cannot be combined across sections and subsections. Therefore, the sections and subsections do not describe separate embodiments.

[0211] All references, including patent applications, patent publications, and UniProtKB / Swiss-Prot accession numbers, cited herein are hereby incorporated by reference in their entirety to the same extent as if each individual reference was specifically and individually indicated to be incorporated by reference.

[0212] 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. It should also be understood that the term "and / or" used herein refers to and encompasses any and all possible combinations of one or more of the relevant listed items. It should also be further understood that when the terms "include" and / or "comprises" are used herein, they specify the presence of specified 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.

[0213] As used herein, the term "about" is meant to include common error ranges for readily known corresponding values. References herein to "about" a value or parameter include (and describe) embodiments involving the value or parameter itself. For example, a description referring to "about X" includes a description of "X". In some embodiments, "about" may refer to ±15%, ±10%, ±5%, or ±1% as understood by those skilled in the art.

[0214] As used herein, the term "amino acid" generally refers to any organic compound containing an amino group (eg, -NH2), a carboxyl group (-COOH), and a side chain group (R) that varies based on the specific amino acid. Amino acids can be linked using peptide bonds.

[0215] As used herein, the phrase "at least one of..." when used with a list of items means that different combinations of one or more of the listed items can be used, and only one item in the list may be needed. An item can be a specific object, thing, step, operation, process, or category. In other words, "at least one of..." refers to any combination of items or number of items that can be used in the list, but not all items in the list are required. For example, but not limited to, "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" can mean, but not limited to, two items in item A, one item in item B, and ten items in item C; four items in item B and seven items in item C; or some other suitable combination.

[0216] Although the present disclosure supports definitions referring only to alternatives and "and / or", the use of the term "or" in the claims is intended to mean "and / or" unless explicitly stated to refer only to alternatives or the alternatives are mutually exclusive. For example, "x, y, and / or z" may 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 expressly contemplated that x, y, or z may be explicitly excluded from an embodiment.

[0217] The term "biological agent" as used herein generally refers to a biological molecule, such as a protein or chemical produced by a cell or virus. The term can refer to any molecule, such as, but not limited to, a protein, a peptide, an antibody (including antibody derivatives, such as, but not limited to, Fc fusions, scFv, multispecific antibodies, bispecific antibodies, etc.), a nucleic acid, a metabolite, an antigen, a chemical substance produced by a cell or a virus, or a biopharmaceutical. The term can also refer to an adeno-associated virus (AAV), for example, an AAV-based gene therapy vector produced by a cell. In some embodiments, the biological agent is easily and / or can be easily made suitable for administration to a subject. In some embodiments, the biological agent is purified and / or formulated before making it suitable for administration to a subject.

[0218] As used herein, the terms "biological sample", "biological specimen" or "biospecimen" generally refer to a specimen obtained by sampling to represent the source of the specimen, usually from a subject. A biological sample may represent a whole organism, a specific tissue, a cell type, or a class or subclass of interest. A biological sample may contain macromolecules. A biological sample may contain small molecules. A biological sample may contain viruses. A biological sample may contain cells or derivatives of cells. A biological sample may contain organelles. A biological sample may contain a nucleus. A biological sample may contain rare cells from a cell population. A biological sample may contain any type of cell, including but not limited to prokaryotic cells, eukaryotic cells, bacteria, fungi, plants, mammalian or other animal cell types, mycoplasmas, normal tissue cells, tumor cells, or any other cell type, whether derived from a single cell or multicellular organism. A biological sample may contain components of a cell. A biological sample may contain nucleotides (e.g., ssDNA, dsDNA, RNA), organelles, amino acids, peptides, proteins, carbohydrates, glycoproteins, or any combination thereof. The biological sample may comprise a matrix (e.g., a gel or polymer matrix) including cells or one or more components from cells (e.g., cell beads), such as DNA, RNA, organelles, proteins from cells, or any combination thereof. The biological sample may be obtained from a tissue of a subject. The biological sample may comprise hardened cells. Such hardened cells may or may not comprise a cell wall or a cell membrane. The biological sample may comprise one or more components of a cell, but may not comprise other components of the cell. Examples of such components may comprise a cell nucleus or an organelle. The biological sample may comprise living cells. The living cells may be capable of being cultured.

[0219] Throughout the specification, unless the context requires otherwise, the words "comprise / comprises / comprising" will be understood to imply the inclusion of the steps or elements or groups of steps or elements, but not to exclude any other steps or elements or groups of steps or elements. "Consisting of" means including and limited to anything after "consisting of". Thus, the phrase "consisting of" indicates that the listed elements are required or mandatory, and no other elements may be present. "Consisting essentially of" means including any element listed in the phrase, and is limited to other elements that do not interfere with or contribute to the activity or effect specified for the listed elements in the present disclosure. Thus, the phrase "consisting essentially of" indicates that the listed elements are required or mandatory, but no other elements are optional and may or may not be present, depending on whether they affect the activity or effect of the listed elements.

[0220] Furthermore, when the terms "coupled to ...", "communicating" or "communicatively coupled to ..." or similar terms are used herein, a component may be able to communicate directly, indirectly, or simultaneously with another component via one or more wired communication links, one or more wireless communication links, one or more optical communication links, or a combination thereof. Additionally, where a list of elements is referred to (e.g., elements a, b, c), such reference is intended to include any one of the listed elements alone, any combination of less than all of the listed elements, and / or combinations of all of the listed elements.

[0221] As used herein, "contaminants" may refer to components that are considered to be undesirable in cell culture fluids and / or cell culture media. For example, in some cases, contaminants may be, but are not limited to, unintended cells and / or viruses, cell culture debris, cell products that are not the target product of therapeutic interest, cell metabolites, and the like.

[0222] As used herein, "internal standard" can refer to a component that can be included (e.g., incorporated) in the same sample as the target. An internal standard can be used for calibration purposes. Additionally, an internal standard can be used in the systems and methods described herein.

[0223] References throughout this specification to "one embodiment," "an embodiment," "a particular embodiment," "a related embodiment," "an embodiment," "an additional embodiment," or "a further embodiment," or combinations thereof, mean that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the above phrases appearing 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.

[0224] As used herein, a "feed stream" is a stream of fluid that leaves a bioreactor, which may include cells, cell culture media, biological agents, etc., and which is directed to a filter. Typically, the feed stream includes a lower concentration of cells than the retentate stream. In some cases, when no permeate is drawn from the filter, the two streams may have the same cell concentration.

[0225] As used herein, the term "harvest permeate stream" includes fluid that passes through the filter membrane and leaves the filter, which may include cell culture medium, biological preparations, etc., and it is typically directed for further processing. Typically, the harvest permeate stream is active during the harvest process and includes a higher concentration of biological preparations and a lower concentration of cells than the retentate stream.

[0226] As used herein, a "model" may include one or more algorithms, one or more mathematical techniques, one or more machine learning algorithms, or a combination thereof.

[0227] As used herein, the term "multi-use module" generally refers to an apparatus comprising at least two flow paths, wherein each flow path is operably connected to a pump configured to withdraw fluid from the permeate stream of a cell retention device at a desired rate.

[0228] As used herein, the term "ones" means more than one.

[0229] As used herein, the term "plurality" may be 2, 3, 4, 5, 6, 7, 8, 9, 10 or more.

[0230] The term "peptide" as used herein generally refers to amino acids connected by peptide bonds. A peptide may comprise an amino acid chain between 10 and 50 residues. A peptide may comprise an amino acid chain shorter than 10 residues, including oligopeptides, dipeptides, tripeptides, and tetrapeptides. A peptide may comprise a chain longer than 50 residues, and may be referred to as a "polypeptide" or "protein."

[0231] As used herein, the term "perfusion permeate stream" includes fluid that passes through the filter membrane and exits the filter, which may include cell culture media, biologics, etc., and which is typically directed for further processing and / or disposal. Typically, the perfusion permeate stream is active during the perfusion process and includes a higher concentration of cell culture media and a lower concentration of biologics than the harvest permeate stream.

[0232] As used herein, the term "permeate" includes fluid that passes through the filter membrane and exits the filter.

[0233] The terms "protein" or "polypeptide" or "peptide" are used interchangeably herein and generally refer to a molecule comprising at least three amino acid residues. A protein may comprise a polymer chain consisting of a sequence of amino acids linked together by peptide bonds. Proteins may be digested using a trypsin digestion protocol in preparation for mass spectrometry analysis. If access is limited to the cleavage site, other proteases may be used to digest the protein in preparation for mass spectrometry analysis.

[0234] As used herein, the term "retentate stream" includes fluid that leaves the filter without passing through the filter membrane (e.g., the retentate stream passes through the filter lumen), which may include cells, cell culture media, biological preparations, etc., and the retentate stream is usually directed to a bioreactor. Typically, the retentate stream includes a higher concentration of cells than the feed stream. In some cases, when no permeate is drawn from the filter, the two streams may have the same cell concentration.

[0235] The term "sample" as used herein generally refers to a sample 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 may include one or more cells. The sample may include 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 cheek swab. The sample may include a plasma or serum sample. The sample may include a cell-free or non-cellular sample. A cell-free sample may include 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 stool, cerebrospinal fluid, CNS fluid, gastric fluid, amniotic fluid, cystic fluid, peritoneal fluid, bone marrow, bile, other body fluids, tissue obtained from a biopsy, skin, or hair.

[0236] As used herein, the term "a set" refers to one or more. For example, a set of items includes one or more items.

[0237] As used herein, the term "sequence" generally refers to a biological sequence comprising one-dimensional monomers that can be assembled to produce 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 carbohydrate sequences (e.g., comprising C m (H2O) n of compounds).

[0238] The term "subject" as used herein generally refers to an animal, such as a mammal (e.g., a human) or a bird (e.g., a bird) or other organism, such as a plant. For example, a subject may include a vertebrate, a mammal, a rodent (e.g., a mouse), a primate, a monkey, or a human. Animals may include, but are not limited to, farm animals, racing animals, and pets. A subject may include a healthy or asymptomatic individual, an individual suffering from or suspected of suffering from a disease (e.g., cancer) or susceptible to the disease, and / or an individual in need of treatment or suspected of needing treatment. A subject may be a patient. A subject may include a microorganism (microbe) (e.g., bacteria, fungi, archaea, viruses).

[0239] As used herein, "substantially" means sufficient to achieve the intended purpose. Thus, the term "substantially" allows for minor, inconsequential variations relative to an absolute or perfect state, dimension, measurement, result, etc., as would be expected by one of ordinary skill in the art, but without significantly affecting the overall performance. When applied to a numerical value or a parameter or characteristic that can be expressed as a numerical value, "substantially" means within ten percent.

[0240] The following description provides only exemplary embodiments and is not intended to limit the scope, applicability or configuration of the present disclosure. On the contrary, the following description of the preferred exemplary embodiments will provide a feasible description for implementing various embodiments for those skilled in the art. It should be understood that various changes may be made to the functions and arrangements of the elements without departing from the spirit and scope set forth in the appended claims.

[0241] Specific details are given in the following description to provide a thorough understanding of the embodiments. However, it should be understood that the embodiments can be practiced without these specific details. For example, circuits, systems, networks, processes, and other components can be shown as parts in block diagram form to avoid obscuring the embodiments in unnecessary details. In other cases, well-known circuits, processes, algorithms, structures, and techniques can be shown without unnecessary details to avoid obscuring the embodiments.

[0242] III. Overview of Multi-Purpose Components

[0243] Referring to FIG. 1 , a non-limiting schematic diagram of an exemplary multi-purpose assembly 100 for improved production of biologics is shown. The assembly includes a feed flow conduit 101 operably connected to an inlet of a cell retention device, such as a hollow fiber (HF) filter 102. In certain embodiments, the feed flow conduit can be operated with an optional feed pump 121. In some embodiments, the HF filter is operable for tangential flow. In some embodiments, the HF filter is a tangential flow depth filtration® (TFDF®) filter. The assembly is operable to facilitate entry of a fluid (e.g., a fluid comprising a cell culture medium, cells, a biologic, etc.) into the HF filter while allowing appropriate fluid components (e.g., those of sufficient size to do so) to pass through the filter membrane and leave the HF filter as a permeate 103. The HF filter is operable so that fluid that does not pass through the filter membrane can leave the HF filter as a retentate 112. The permeate fluid can be pulled through the HF filter to enter two or more flow paths by the action of two or more pumps. Under certain operating conditions, such as operating during a perfusion process, the first pump 106 is operable to pull the permeate 103 into the first flow path 104, thereby forming a perfusion permeate 107. The first pump 106 is operable to communicate with a sensor 105, which is operable to accurately measure the fluid flow rate, so that the first pump 106 is operable to maintain a fluid flow rate suitable for the perfusion process, for example, wherein the perfusion permeate 107 from the HF filter is pulled at a consistent and / or desired rate. Under certain operating conditions, such as operating during a harvest process, the second pump 110 is operable to pull the permeate 103 into the second flow path 108, thereby forming a harvest permeate 111. The second pump 110 and the harvest permeate 111 are optionally operable to communicate with one or more sensors and / or in fluid communication with one or more downstream filtration units and / or processing units suitable for maintaining the harvest process. Typically, the second flow path 108 and the second pump 110 draw permeate at a higher fluid flow rate than the first flow path 104 and the first pump 106 .

[0244] In certain embodiments, the multi-purpose assembly is operable for one or more perfusion processes (such as, but not limited to, 1, 2, 3, 4, 5, 6, or more than 6 perfusion processes) and one or more harvesting processes (e.g., a harvesting process and / or an additional process after the harvesting process begins). 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, during which permeate may or may not be extracted and fluid flow may or may not be maintained), and one or more harvesting processes (e.g., an additional process after the harvesting process begins). In some embodiments, such a combination of processes is referred to as a combined process.

[0245] In certain embodiments, the multi-purpose assembly is connected to an adjustable bioreactor. In certain embodiments, the adjustment ratio is about 30:1 to about 2:1. In certain embodiments, the adjustment ratio is about 20:1 to about 5:1. In certain embodiments, the adjustment ratio is 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 adjustment 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 adjustment ratio of about 5:1.

[0246] In certain embodiments, more than one multi-purpose assembly can be used in a system. In certain embodiments, one, two, three, four, or five multi-purpose assemblies can be used in a system. In certain embodiments, when more than one multi-purpose assembly is used in a system, the assemblies are used sequentially and / or in series. In certain embodiments, using more than one multi-purpose assembly in a system including a bioreactor can allow for an increase in bioreactor volume relative to a system having only one multi-purpose assembly.

[0247] In certain embodiments, exemplary operating parameters for three target throughputs achievable using the multi-purpose assembly and / or systems including the multi-purpose assembly are provided in Table 1.

[0248] Table 1 - Exemplary Perfusion Process Operating Parameters

[0249]

[0250] III.A. Cell Retention Device (CRD)

[0251] In certain embodiments, the CRD is a filter suitable for both perfusion and harvesting processes. In certain embodiments, the CRD is a hollow fiber (HF) filter. In certain embodiments, the HF filter is operable for 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™), NÜF N80 (Ochemate®), or a Tangential Flow Depth Filtration® (TFDF®; Repligen® Inc.) filter. In certain embodiments, the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter. In certain embodiments, the HF filter is composed of PES / PVDF, PEI-based TFC, PES-PEM, mPES, and / or polyamide-based TFC.

[0252] In some embodiments, the HF filter consists of and / or includes a synthetic polymer. In some embodiments, the HF filter consists of and / or includes polypropylene and / or polyethylene terephthalate. In some embodiments, the HF filter consists of and / or includes modified polyethersulfone, polyethersulfone, mixed cellulose esters and / or polysulfone.

[0253] In certain embodiments, the HF filter has a range from about 3 to about 6240 cm 2 In certain embodiments, the HF filter has a filter surface area of ​​about 3 cm 2 , about 30 cm 2 , about 50 cm 2 , about 150 cm 2 、Approx. 450 cm 2 、Approx. 750 cm 2 、Approx. 1,500 cm 2 、Approx. 2,100 cm 2 、Approx. 6,000 cm 2 or greater than about 6,000 cm 2In 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 more than 9,000 cm 2 filter surface area.

[0254] In some embodiments, the HF filter comprises one or more fibers. In some embodiments, the fiber has an inner cavity through which a feed stream can flow, surrounded by a membrane layer through which a permeate can pass. In some embodiments, a permeate fluid drawn through the membrane layer can exit the HF filter as a permeate (see, e.g., FIG. 1 , permeate 103). In some embodiments, the length of the tube is 20, 36, or 108 cm. In some embodiments, the length of 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, 122, 123, 124, 125, 126, 127, 128, 129, 130 or more than 130 cm.

[0255] In some embodiments, the HF filter comprises more than 1 fiber. 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, 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, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209

[0256] In some embodiments, the HF filter has an average pore size between about 0.65 μm and about 8 μm. In some embodiments, the HF filter has an average pore size between about 2 μm and about 5 μm. In some embodiments, the HF filter has an average pore size of about 2 μm. In some 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 range derivable therein. In some embodiments, the pore structure is anisotropic. In some embodiments, the pore structure is not anisotropic. In some embodiments, the pore structure is isotropic.

[0257] In certain embodiments, the HF filter filtrate channel comprises a filter having an inner lumen of about 2 mm passing therethrough. In certain embodiments, the HF filter filtrate channel comprises a filter having an inner lumen of about 4.6 mm passing therethrough. In certain embodiments, the TFDF® filter filtrate channel comprises a filter having an inner 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 therein, passing therethrough. In certain embodiments, the HF filter filtrate channel comprises a filter having an inner lumen of about 0.4 mm to about 1.5 mm passing therethrough. In certain embodiments, the HF filter filtrate channel does not have an inner lumen of less than about 0.4 mm passing therethrough.

[0258] In certain embodiments, the HF filter has a wall thickness of about 0.05 mm to about 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 of 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.

[0259] In certain embodiments, the permeate may flow through the HF filter at a flow rate of 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 may 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 therein. In certain embodiments, the permeate may flow through the HF filter at a flow rate between about 0 LPM and / or about 0.1 LPM to about 5 LPM. In certain embodiments, the permeate may flow through the HF filter at a flow rate between about 0 LPM and / or about 0.1 LPM to about 8 LPM. In certain embodiments, the permeate may flow through the HF filter at a flow rate between about 0 LPM and / or about 0.1 LPM to about 10 LPM. In certain embodiments, the permeate may flow through the HF filter at a flow rate between about 0 LPM and / or about 0.1 LPM to about 13 LPM. In certain embodiments, the permeate may flow through the HF filter at a flow rate between about 0 LPM and / or about 0.1 LPM to about 15 LPM. In certain embodiments, the permeate may flow through the HF filter at a flow rate between about 0 LPM and / or about 0.1 LPM to about 18 LPM.

[0260] In certain embodiments, the permeate may 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.

[0261] In certain embodiments, the HF filter has a density 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, 0, 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, 6 0,000, 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 L / m 2 ) . In some embodiments, the HF filter has a throughput of greater than about 10,000 L / m 2 In some embodiments, the HF filter has a throughput of greater than about 25,000 L / m 2 In some embodiments, the HF filter has a throughput of greater than about 30,000 L / m 2 In some embodiments, the HF filter has a throughput of greater than about 40,000 L / m 2 In some embodiments, the HF filter has a flow rate of about 5,000 to about 35,000 L / m 2 The throughput of any range between or derivable therein.

[0262] In certain embodiments, the HF filter has a filter flux rate between about 20 liters per square meter per hour and about 800 liters per square meter per hour (LMH). In certain embodiments, the HF filter has a filter flux rate between about 100 and about 600 LMH. In certain embodiments, the HF filter has a filter flux rate between about 300 and about 600 LMH. In certain embodiments, the HF filter has a filter flux rate between about 400 and about 600 LMH. In certain embodiments, the HF filter has a filter flux rate between about 50 and about 100, about 50 and about 150, about 50 and about 200, about 50 and about 250, about 50 and about 300, about 50 and about 350, about 50 and about 400, about 50 and about 450, about 50 and about 500, about 50 and about 550, about 50 and about 600, about 50 and about 650, about 50 and about 700, about 50 and about 750, about 50 and about 800, about 50 and about 850, or about 50 to about 900 LMH.

[0263] In some embodiments, the multi-use filter assembly is operable to maintain a volume of greater than about 1 x 10 6 In some embodiments, the multi-purpose filter assembly is operable to maintain a viable cell density (VCD) of greater than about 10 x 10 6 In some embodiments, the multi-purpose filter assembly is operable to maintain a viable cell density (VCD) of greater than about 25 x 10 6 In some embodiments, the multi-purpose filter assembly is operable to maintain a viable cell density (VCD) of greater than about 50 x 10 6 In some embodiments, the multi-use filter assembly is operable to maintain a VCD of greater than about 75 x 10 6 In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD of greater than about 10 x 10 7 In some embodiments, the multi-use filter assembly is operable to maintain a VCD of greater than about 25 x 10 7 In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD of greater than about 50 x 10 7 In some embodiments, the multi-use filter assembly is operable to maintain a VCD of greater than about 75 x 107 In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD of greater than about 10 x 10 8 In some embodiments, the multi-use filter assembly is operable to maintain a VCD of greater than about 25 x 10 8 In some embodiments, the multi-purpose filter assembly is operable to maintain a VCD of greater than about 50 x 10 8 In some embodiments, the VCD is the VCD in the bioreactor fluid volume. In some embodiments, the VCD is the VCD in the retentate stream of the CRD.

[0264] III.B. Pumps

[0265] In some embodiments, the assemblies, systems, and / or methods described herein include or comprise the use of one or more pumps.

[0266] In some embodiments, the pump can be operable to push or pull fluid through a suitable conduit. In some embodiments, the pump can be, but is not limited to, a pump selected from the group consisting of a peristaltic pump, a centrifugal pump, a magnetically driven pump, a positive displacement pump, a diaphragm pump, a pressure-based pump, a quantex™ pump (e.g., a positive displacement rotary pump), a gear pump, a diaphragm pump, a syringe pump, and a piston pump. In some embodiments, the pump is a magnetically driven pump. In some embodiments, the pump is a peristaltic pump. In some embodiments, one or more pumps are suitable for generating alternating tangential flows. In some embodiments, one or more pumps are not used for and / or are not operable to generate alternating tangential flows.

[0267] In some embodiments, the pump is operable to maintain a desired flow rate. In some embodiments, the pump communicates with one or more sensors that can direct the pump to increase or decrease pumping force to change the flow rate as desired. In some embodiments, the flow rate is measured in liters per fiber per minute (L / fiber / min). In some embodiments, the flow rate is or is 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 L / fiber / min. In some embodiments, the flow rate is about 0.5 to about 2.5 L / fiber / min. In some embodiments, the flow rate is about 0.8 to about 2.2 L / fiber / min. In some embodiments, the flow rate is about 0.8 to about 2.0 L / fiber / min. In some embodiments, the flow rate is about 1 to about 1.8 L / fiber / min.

[0268] In some embodiments, the flow rate is determined according to a set container volume per day (VVD). 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 used in conjunction with a relatively small bioreactor, the pump can maintain a higher VVD rate.

[0269] In some embodiments, the system may include more than one multipurpose component. In some embodiments, where the system includes more than one multipurpose component, the VVD rate may be scaled accordingly, such as but not limited to about 5, 6, 7, 8, 9, 10 or more than 10 VVD.

[0270] In some embodiments, the pump is operable to accurately facilitate fluid flow at a rate of 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 some embodiments, the pump is operable to accurately facilitate the flow of the liquid at a rate of 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, 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 7, 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, 59 77.5, 78, 78.5, 79, 79.5 or 80 LPM or any range of flow rates derivable therein.

[0271] In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate ranging from about 0 LPM and / or about 0.1 LPM to about 20 LPM or any range derivable therein. In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate ranging from about 0 LPM and / or about 0.1 LPM to about 5 LPM. In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate ranging from about 0 LPM and / or about 0.1 LPM to about 8 LPM. In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate ranging from about 0 LPM and / or about 0.1 LPM to about 10 LPM. In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate ranging from about 0 LPM and / or about 0.1 LPM to about 13 LPM. In some embodiments, the pump is operable to accurately facilitate the flow of fluid at a flow rate ranging 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 ranging from about 0 LPM and / or about 0.1 LPM to about 18 LPM.

[0272] In some embodiments, the first flow path pump is operable to accurately facilitate fluid flow at a rate less than 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 the pump associated with the harvest permeate flow path.

[0273] In some embodiments, a first flow path pump associated with a precision sensor is adapted to precisely control fluid flow in the first flow path. In some embodiments, a first flow path pump associated with a precision sensor is adapted to precisely control fluid flow in the first flow path, wherein the control is more accurate than a pump used to direct flow in the second flow path.

[0274] In some embodiments, the pump can have a head of any size. In some embodiments, the pump head is sized according to the desired flow rate, the inner diameter of the tubing, and / or the flow accuracy requirements. In some embodiments, the tubing in the flow path can be of any type suitable for cell culture. In some embodiments, the tubing can be autoclavable. In some embodiments, the tubing is suitable for food grade quality and / or good manufacturing purpose quality. In some embodiments, the tubing comprises silicone.

[0275] In some embodiments, the pump is operable to maintain a PCV percentage 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 therein. 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%.

[0276] III.C Sensor

[0277] The multi-purpose assembly includes at least one sensor adapted to accurately measure fluid flow in a first flow path (see, e.g., FIG. 1 , flow path 104). In some embodiments, such sensor is in communication with a first flow path pump. In some embodiments, such sensor is in communication with a second flow path pump.

[0278] In some embodiments, the sensor is operable to accurately measure fluid flow at a flow rate of 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).

[0279] In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate ranging from about 0 LPM and / or about 0.1 LPM to about 20 LPM or any range derivable therein. In some embodiments, the sensor is operable to accurately measure the flow of fluid at a flow rate ranging from 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 ranging from 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 ranging from 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 ranging from 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 ranging from about 0 LPM and / or about 0.1 LPM to about 15 LPM. In some embodiments, the sensor is operable to accurately measure fluid flow at flow rates ranging from about 0 LPM and / or about 0.1 LPM to about 18 LPM.

[0280] In some embodiments, at least one sensor adapted to accurately measure fluid flow in the first flow path is more accurate than one or more additional sensors adapted to measure flow in the second flow path.

[0281] 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.

[0282] III.D flow path

[0283] As described herein, in most embodiments, the multi-purpose assembly includes at least two flow paths. In certain embodiments, the multi-purpose assembly includes 2 flow paths. In certain embodiments, the multi-purpose assembly includes more than 2 flow paths, such as but not limited to 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 flow paths or any range derivable therein. In some embodiments, flow paths other than the first two flow paths may be specifically operable for certain processes (e.g., certain perfusion and / or harvesting processes) or may be generalized as operable for any process requiring a flow path.

[0284] In certain embodiments, the first flow path is a perfusion permeate flow path (see, e.g., FIG. 1 , first flow path 104 and perfusion permeate 107). In certain embodiments, the first flow path comprises tubing having an inner diameter (ID) of 1 / 2", 3 / 8", 1 / 4", or 1 / 8". In certain embodiments, the first flow path comprises tubing having a 3 / 8" ID.

[0285] In certain embodiments, the second flow path is a harvest permeate flow path (see, e.g., FIG. 1 , flow path 108 and harvest permeate 111). In certain embodiments, the second flow path comprises tubing having an inner diameter (ID) of 1 / 2", 3 / 8", 1 / 4", or 1 / 8". In certain embodiments, the second flow path comprises tubing having a 1 / 2" ID.

[0286] In certain embodiments, a multi-purpose assembly including a first flow path and a pump is operable to extract a flow rate of about 0 liters per fiber per minute to about 3 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 extract 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 extract 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 extract a flow rate of about 1.0 to about 1.8 L / fiber / min from the HF filter.

[0287] In certain embodiments, the multi-purpose assembly including the second flow path and the pump is operable to extract a flow rate of about 0 liters per minute per fiber to about 4 liters per minute per fiber (L / fiber / min) from the HF filter. In some embodiments, the second flow path and the pump are operable to extract 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 extract 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 extract a flow rate of about 1.5 to about 2 L / fiber / min from the HF filter.

[0288] 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 flow rate between about 0.01 LPM to about 80 LPM, between about 0.01 LPM to about 60 LPM, between about 0.01 LPM to about 40 LPM, and / or between about 0.01 LPM to about 20 LPM.

[0289] In some embodiments, the multi-purpose assembly including the second flow path and the pump is operable to pump at a rate of greater than or equal to 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 approximately 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 In some embodiments, the multi-purpose assembly including the second flow path and the pump is operable to extract fluid from the HF filter at a throughput of greater than or equal to about 4,000 liters per square meter (L / m 2 ) of the HF filter. In some embodiments, the multipurpose assembly including the second flow path and the pump is operable to pump fluid at a rate of greater than or equal to about 40,000 liters per square meter (L / m 2 ) of the throughput to draw fluid from the HF filter.

[0290] 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 may be a flow accuracy requirement.

[0291] In some embodiments, the first flow path and the second flow path are connected to the primary permeate flow path (see, e.g., FIG. 1 , flow path 103 and first flow path 104 and second flow path 108). In some embodiments, the first flow path and the second flow path 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, an optional clamp (e.g., a clamp) may be operable to direct flow to the first flow path or the second flow path. In some embodiments, the opening and closing of the valve may be automated.

[0292] 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 lower than the fluid flow rate through the harvesting flow path.

[0293] III.E Filters

[0294] In some embodiments, the multi-use assembly can include one or more additional filters (e.g., clarification filters). In some embodiments, the additional filters are attached to the first flow path. In some embodiments, the additional filters are attached to the second flow path. The additional filters can be configured to, but are not limited to, remove contaminants, selectively remove and / or harvest one or more biological agents, and / or to ensure the sterility of the closed system.

[0295] III.F. Control unit and human-machine interface

[0296] In some embodiments, a multi-purpose assembly is provided herein 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 can be used to monitor and / or control any adjustable parameters associated with the multi-purpose assembly, such as but not limited to VVD, flow, VCD, pump speed, sensor accuracy, flow path utilization, etc. In some embodiments, the control unit and HMI can be used to control additional parameters associated with a system including the multi-purpose assembly, such as but not limited to temperature, fresh medium input rate, discharge rate, inducer addition rate, oxygen level, CO2 level, metabolite level, etc.

[0297] In some embodiments, a customized control strategy is programmed, integrated, and / or displayed using an HMI. In some embodiments, any physical components and / or devices described herein may be operable for and / or communicate with an HMI.

[0298] IV. Overview of Systems Including Multi-Purpose Components

[0299] Referring to FIG2 , a non-limiting schematic diagram of an exemplary bioreactor system 200 is shown, which includes a multi-purpose assembly for improving the production of biologics (see, e.g., FIG1 , 100 ). The system includes a bioreactor 220 operably connected to an optional inlet (e.g., fresh medium input port) 223, an output port (e.g., feed stream) 201, and an input (e.g., retentate) port 212. The system may include a bioreactor fluid volume 222. The system includes 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 for tangential flow. In some embodiments, such HF filters are tangential flow depth filtration® (TFDF®) filters. The output port 201 may optionally include a pump 221 for facilitating fluid flow from the bioreactor to the HF filter 202. The system is operable to facilitate fluid (e.g., fluid including cell culture media, cells, biologics, etc.) into the HF filter while allowing appropriate fluid components (e.g., those of sufficient size to do so) to exit the HF filter through the filter membrane as permeate 203. The HF filter is operable so that fluid that does not pass through the filter membrane can exit the HF filter through an outlet as retentate 212 to be reintroduced into the bioreactor 220. The permeate fluid can be pulled through the HF filter into two or more flow paths by the action of two or more pumps. Under certain operating conditions, such as operating during a perfusion process, a first pump 206 is operable to pull the permeate 203 into the first flow path 204, thereby forming a perfusion permeate 207. The first pump 206 is operable to communicate with a sensor 205, which is operable to accurately measure the fluid flow rate, so that the first pump 206 is operable to maintain a fluid flow rate suitable for the perfusion process, for example, wherein the perfusion permeate 207 from the HF filter is pulled at a consistent and / or desired rate. Under certain operating conditions, such as operation during a harvesting process, the second pump 210 is operable to draw the permeate 203 into the second flow path 208, thereby forming a harvested permeate 211. The second pump 210 and the harvested permeate 211 are optionally operable to communicate with one or more sensors and / or to communicate with one or more downstream filtration units and / or processing units suitable for maintaining the harvesting process. Typically, 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.

[0300] IV.A. Bioreactor

[0301] In some embodiments, systems including multi-purpose components are described herein. In some embodiments, such systems can be operated with a bioreactor. A bioreactor is a container operable to support a biologically active environment. The bioreactor can be suitable for supporting aerobic and / or anaerobic biological activity. In some embodiments, the bioreactor can be a batch bioreactor, a feed batch bioreactor, a continuous bioreactor, a semi-continuous bioreactor or a perfusion bioreactor. In some embodiments, the bioreactor can have any size. In some embodiments, the bioreactor can be suitable for cell growth and / or maintenance during any stage of cell culture (e.g., a slow phase, a logarithmic phase, a stationary phase, a death phase, etc.). In some embodiments, the bioreactor can have 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 can be operable to be connected to more than one multi-purpose filter assembly as described herein. In some embodiments, wherein the bioreactor can be operated with more than one multi-purpose filter assembly, such more than one multi-purpose filter assembly can act simultaneously or sequentially. In some embodiments, more than one bioreactor container can be used in series or sequentially.

[0302] In some embodiments, the bioreactor container may include greater than or equal to 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 In some embodiments, the bioreactor vessel may include a volume of less than or equal to about 15 liters. In some embodiments, the bioreactor vessel may include a volume of less than or equal to about 50 liters. In some embodiments, the bioreactor vessel may include a volume of greater than or equal to about 3,000 liters. In some embodiments, the bioreactor vessel may include a volume of greater than or equal to about 6,000 liters.

[0303] In some embodiments, the total volume of the bioreactor vessel is greater than the working volume of the cell culture fluid included in the bioreactor. In some embodiments, the total volume of the bioreactor vessel can be greater than or equal to 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 therein.

[0304] In some embodiments, the bioreactor vessel is maintained at a set temperature. In some embodiments, the bioreactor vessel is maintained at about 20°C to about 40°C, about 30°C to about 38°C, or about 35°C to about 38°C. In certain embodiments, the bioreactor vessel is maintained at about 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, or 43°C. In certain embodiments, the bioreactor vessel is maintained at about 37°C.

[0305] IV.B. Cell Culture Medium

[0306] In some embodiments, described herein are systems and / or components that may include cell culture fluids. In some embodiments, the cell culture fluid may be any cell culture fluid known in the art. In some embodiments, the cell culture fluid may include cells, biological preparations, cell culture media, metabolites, etc. In some embodiments, the feed stream of the cell culture media is added to the cell culture fluid. In some embodiments, the cell culture media may include concentrated amounts of cell nutrients, such as sugars, peptides, vitamins, lipids, etc. In some embodiments, fresh cell culture media is introduced into the bioreactor as shown in Figure 2 via an optional input port 223. In some embodiments, the permeate cell culture media includes metabolites and / or biological preparations at lower concentrations than the cell culture fluid and / or fresh cell culture media. In some embodiments, the harvested permeate cell culture media includes biological preparations at higher concentrations than the perfusion permeate cell culture media.

[0307] V. How to use

[0308] Referring to FIG. 3 , a non-limiting exemplary flow chart is shown, which illustrates a method for producing a biologic using an HF filter capable of operating during both a perfusion process and a harvest process. In certain embodiments, the HF filter is operable for tangential flow. In certain embodiments, the HF is a TFDF® filter. Method 300 may include: at step 302, a cell population capable of producing a biologic and / or capable of promoting biologic production (e.g., capable of amplifying a viral population, acting as a trophoblast, etc.) in a cell culture fluid in a bioreactor. During step 304, cells are amplified using a perfusion process cell culture technique, wherein the cell culture fluid passes through the HF filter to remove spent cell culture medium (e.g., the culture medium is perfused through the HF filter membrane), while the cells remain in the HF filter lumen and are circulated back to the bioreactor through the retentate flow. During the perfusion phase, spent cell culture medium is perfused through the HF filter at a first flow rate to enter a first flow path (e.g., perfusion permeate flow path). In certain embodiments, an appropriate replacement volume of cell culture medium is added to the bioreactor to maintain the desired cell culture fluid content in the bioreactor. In certain embodiments, the replacement volume of the cell culture medium is approximately equal to, less than, or greater than the volume of the spent cell culture medium removed. In certain embodiments, the replacement volume of the cell culture medium is fresh cell culture medium. During step 306, after the cell expansion is performed using the perfusion process, the harvest process is started. In certain embodiments, between the perfusion process and the harvest process, an intermediate process is performed. During such a harvest process, the cell culture medium is perfused through the HF filter at a second flow rate to enter the second flow path (e.g., the harvest permeate flow path). In certain embodiments, an appropriate replacement volume of the cell culture medium is added to the bioreactor to maintain the desired cell culture fluid content in the bioreactor during the different stages of the harvest. In certain embodiments, the replacement volume of the cell culture medium is approximately equal to, less than, or greater than the volume of the spent cell culture medium removed. In certain embodiments, the replacement volume of the cell culture medium 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 a specific embodiment, the first flow rate in the first flow path is less than or equal to about 5 LPM, and the second flow rate in the second flow path is greater than about 5 LPM. In some embodiments, the method includes using any system or component described herein.

[0309] In some embodiments, the systems and apparatus described herein can be used in various processing schemes, such as, but not limited to: perfusion, batch, fed-batch, semi-continuous processing (eg, continuous harvest, extended harvest, etc.), and / or continuous processing.

[0310] In some embodiments, the systems and devices described herein are used in methods involving large liquid volumes, high liquid flow rates, high throughput, and / or high pressures, and are suitable for use with sensitive cells (e.g., animal cells, e.g., mammalian cells) that are susceptible to shear at large liquid volumes, high liquid flow rates, high throughput, and / or high pressures.

[0311] VA cells

[0312] The components and systems described herein can be used with any type of cell. In certain embodiments, the cells are considered to be relatively sensitive cells (e.g., the cells are shear-sensitive, e.g., the cells are susceptible to shearing at certain large volumes, high flows, high throughput rates, and / or high pressures) when compared to cells such as E. coli or yeast. 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 (iPSC). In certain embodiments, the cells are stem cells and / or iPSCs that have differentiated into hematopoietic lineages. 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 fetal kidney 293 (HEK293) cells. In certain embodiments, the cell is a human fibrosarcoma cell (e.g., HT-1080 cell). In certain embodiments, the cell is derived from immortalized human embryonic cells (e.g., PER.C6 cells). In certain embodiments, the cell is a fusion of HEK293-S and a human B cell line (e.g., HKB-11 cell). In certain embodiments, the cell is derived from a human hepatocellular carcinoma cell (e.g., HuH-7 cell). In certain embodiments, the cell is a trophoblast and / or host cell for producing one or more viruses.

[0313] VB Products

[0314] The components and systems described herein can be used to produce products. In certain embodiments, the products are cells (e.g., stem cells, hematopoietic lineage cells, immune effector cells, etc.). In certain embodiments, the products are biologics. In certain embodiments, the biologics can be of any type. In certain embodiments, the biologics can be, but are not limited to, small molecules, peptides, proteins, antibodies (including antibody derivatives, such as, but not limited to, Fc fusions, scFv, multi-specific antibodies, bispecific antibodies, etc.), carbohydrates, lipids, viruses, virus-like particles, viral proteins / peptides, extracellular particles (e.g., microvesicles, exosomes, etc.), vaccines, and / or nucleotides (e.g., DNA and / or RNA molecules).

[0315] In certain embodiments, the biologic comprises an antibody or a functional unit thereof (e.g., a single-chain variable fragment (scFv), a heavy chain, a light chain, a fragment crystallizable (Fc) domain, etc.), is essentially composed of an antibody or a functional unit thereof, or is composed of an antibody or a functional unit thereof. In certain embodiments, the biologic comprises a viral vector, is essentially composed of a viral vector, or is composed of a viral vector. In certain embodiments, the biologic comprises an adeno-associated virus (AAV), is essentially composed of an adeno-associated virus, or is composed of an adeno-associated virus. In certain embodiments, the biologic comprises a lentivirus, is essentially composed of a lentivirus, or is composed of a lentivirus. In certain embodiments, the biologic comprises an adenovirus, is essentially composed of an adenovirus, or is composed of an adenovirus. In certain embodiments, the biologic does not comprise, is not essentially composed of, or is not composed of AAV, lentivirus, and / or adenovirus. In certain embodiments, the biologic comprises, consists essentially of, or consists of an RNA molecule (such as, but not limited to, messenger RNA (mRNA), small interfering RNA (siRNA), micro RNA (miRNA), long noncoding RNA (lncRNA), and / or antisense oligonucleotides (ASOs)). In some embodiments, the biologic comprises, consists essentially of, or consists of an antigenic molecule (e.g., a molecule that can induce an immune response in a subject).

[0316] VC Automation

[0317] In some embodiments, the components and systems described herein and methods of using them can be automated. In certain embodiments, automation includes integration of one or more human machine interfaces (HMIs) and processing devices. In some embodiments, one or more sensors can provide real-time data that can be displayed on the HMI to the processing device. In some embodiments, the actions required to change the biomanufacturing process (e.g., a perfusion process, an intermediate process, or a harvesting process) can be programmed into the processing device and controlled at the HMI.

[0318] VD Time

[0319] In some embodiments, the components and systems described herein and methods of using the same can last for any period of time. In certain embodiments, the perfusion process comprising the use of a multi-purpose component as described herein can last for equal to or greater than 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 longer than 60 days or any range derivable 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 range derivable therein. In certain embodiments, the perfusion process is about 5 days. In certain embodiments, the perfusion process is about 6 days.

[0320] In certain embodiments, the HF filter is not exchanged between the perfusion process and the harvesting process. In certain embodiments, the harvesting process comprising the use of a multi-use assembly as described herein can last for equal to or greater than 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 longer, or any range derivable therein. In certain embodiments, the perfusion process and the harvesting process comprising the use of the multi-purpose assembly as described herein can last for equal to or greater than 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 longer. Days or any range derivable therein. In certain embodiments, the harvesting 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 greater than about 10 days. In some embodiments, the harvesting process is about 6 days. In some embodiments, the harvesting process is longer than about 6 days. In some embodiments, the harvesting process is a continuous harvesting process.

[0321] VI. Representative Experimental Results

[0322] VI.A. Systems including multi-purpose components

[0323] A multi-use assembly and system including the components depicted in FIGS. 1 and 2 were produced. Biomanufacturing trials were conducted on at least 10 different biological agents, including at least 4 trials that included a combined perfusion and continuous harvesting process. The exemplary multi-use assembly and system including the exemplary multi-use assembly achieved filter flux levels up to 600 LMH, VVD rates up to 4.3, and a flow rate of up to 158 x 10 6 VCD rate and up to 57,500L / m 2The filter pass rate.

[0324] Table 2 - Example experiments, summary of results

[0325]

[0326] Table legend: Perfusion = P; Continuous harvest = CH; Biologics coded; Test site coded.

[0327] VII. Other considerations

[0328] Any headings and / or subheadings between sections and subsections of this document are used only to improve readability and do not mean that features cannot be combined across sections and subsections. Therefore, sections and subsections do not describe separate embodiments.

[0329] Although this teaching is described in conjunction with various embodiments, this teaching is not intended to be limited to such embodiments. On the contrary, this teaching encompasses various substitutions, modifications, and equivalents that will be understood by those skilled in the art. This description provides preferred exemplary embodiments and is not intended to limit the scope, applicability, or configuration of the present disclosure. On the contrary, this description of preferred exemplary embodiments will provide those skilled in the art with a feasible description for implementing various embodiments.

[0330] It should be understood that various changes may be made to the function and arrangement of elements without departing from the spirit and scope set forth in the appended claims. Such modifications and variations are therefore considered to be within the scope specified in the appended claims. Furthermore, the terms and expressions that have been adopted are used as terms of description rather than limitation, and when using these terms and expressions, there is no intention to exclude any equivalents of the features shown and described or parts thereof, but it should be recognized that various modifications are possible within the scope of the invention claimed.

[0331] In describing various embodiments, the specification may have presented the method and / or process as a specific sequence of steps. However, if the method or process does not rely on the specific order of steps described herein, the method or process should not be limited to the specific sequence of steps listed, and those skilled in the art can readily understand that these sequences can be different and still remain within the spirit and scope of the various embodiments.

[0332] Some embodiments of the present disclosure include a system including one or more data processors. In some embodiments, the system includes a non-transitory computer-readable storage medium containing instructions, which, when executed on the one or more data processors, cause the one or more data processors to perform part or all of one or more methods disclosed herein and / or part or all of one or more processes. Some embodiments of the present disclosure include a computer program product tangibly embodied in a non-transitory machine-readable storage medium, which includes instructions configured to cause one or more data processors to perform part or all of one or more methods disclosed herein and / or part or all of one or more processes disclosed herein.

[0333] Specific details are given in this description to understand the present embodiments. However, it should be understood that the embodiments can be practiced without these specific details. For example, circuits, systems, networks, processes, and other components can be shown as parts in block diagram form to avoid obscuring the embodiments in unnecessary details. In other cases, well-known circuits, processes, algorithms, structures, and techniques can be shown without unnecessary details to avoid obscuring the embodiments.

Claims

1. A bioreactor system comprising: a) a bioreactor comprising an input port and an output port; b) a feed flow conduit operably connected to the output port and an inlet of a hollow fiber (HF) filter, the feed flow conduit being operable to convey fluid from the bioreactor to the HF filter; c) a retentate stream conduit operably connected to the input port of the bioreactor and the outlet of the HF filter, the retentate stream conduit being operable to transport 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: wherein the first flow path comprises a first pump operable to extract fluid from the HF filter and a flow meter operable to measure a flow rate of the fluid in the first flow path; wherein the second flow path comprises a second pump operable to draw fluid from the HF filter; and Wherein the first pump and the second pump are configured to have different pump capacities.

2. The bioreactor system of claim 1, wherein the HF filter is operable for tangential flow.

3. The bioreactor system of claim 1 , wherein the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter.

4. The bioreactor system of claim 1, wherein the first pump and the second pump are configured to have different accuracy levels.

5. The bioreactor system according to claim 1, further comprising: One or more clarifying filters in the first flow path and / or the second flow path.

6. The bioreactor system of claim 1 , wherein the first pump and the second pump 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-based pump, a quantex™ pump, a gear pump, a diaphragm pump, a syringe pump, and a piston pump.

7. The bioreactor system of claim 1, wherein the first pump and the second pump are peristaltic pumps.

8. The bioreactor system of claim 1 , wherein the feed flow conduit comprises a feed 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-based pump, a quantex™ pump, a gear pump, a diaphragm pump, a syringe pump, and a piston pump, and is operable to convey unfiltered fluid from the bioreactor to the HF filter.

9. The bioreactor system of claim 1, wherein the retentate stream conduit is operable to transport cell culture fluid from the HF filter to the bioreactor.

10. The bioreactor system of claim 1, wherein the first flow path including the first pump operable to draw fluid from the HF filter draws perfusion permeate flow fluid from the HF filter.

11. The bioreactor system of claim 1, wherein the second flow path including the second pump operable to draw fluid from the HF filter draws a harvest permeate stream fluid from the HF filter. 12 . The bioreactor system of claim 1 , wherein the first flow path and the second flow path have the same inner diameter or have different inner diameters.

13. The bioreactor system of claim 1, wherein the second flow path is capable of conducting fluid at a higher flow rate than the first flow path.

14. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 liters per minute (LPM) and 5 LPM.

15. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM to about 10 LPM.

16. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM to about 15 LPM.

17. The bioreactor system of claim 1, wherein the first flow path and pump are operable to draw a feed flow rate of about 0.8 liters per fiber per minute to about 2.2 liters per fiber per minute (L / fiber / min) from the HF filter.

18. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter while maintaining a flow rate greater than about 75 x 10 6 The viable cell density (VCD) was calculated as cells / mL.

19. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter while maintaining a flow rate greater than about 50 x 10 6 The viable cell density (VCD) was calculated as cells / mL.

20. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter while maintaining a flow rate greater than about 25 x 10 6 cells / mL of VCD.

21. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter at a rate between about 0.1 bioreactor vessel volumes per day and about 5 bioreactor vessel volumes per day (VVD).

22. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter at a rate between about 0.5 bioreactor vessel volumes per day and about 4.3 bioreactor vessel volumes per day (VVD).

23. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter at a filter flux rate of between about 50 liters per square meter per hour and about 800 liters per square meter per hour (LMH).

24. The bioreactor system of claim 1, wherein the first pump is operable to draw fluid from the HF filter at a filter flux rate of between about 100 LMH and about 600 LMH.

25. The bioreactor system of claim 1, wherein the flow meter operable to measure flow of fluid in the first flow path continuously monitors a VVD rate and communicates with the first pump to adjust the first pump rate to maintain a desired VVD rate.

26. The bioreactor system of claim 1, wherein the flow meter operable to measure the flow rate of fluid in the first flow path is capable of accurately monitoring a flow rate of about 0 LPM to about 8 LPM or about 0.5 LPM to about 6 LPM.

27. The bioreactor system of claim 1, wherein the first pump is operable to pump the liquid at a rate equal to about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m2 of fluid is drawn from the HF filter.

28. The bioreactor system of claim 1, wherein the first pump is operable to pump at a rate equal to about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m2 of fluid is drawn from the HF filter.

29. The bioreactor system of claim 1, 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.

30. The bioreactor system of claim 1, 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.

31. The bioreactor system of claim 1, 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.

32. The bioreactor system of claim 1, wherein the bioreactor has a volume less than or equal to about 15 liters or about 50 liters.

33. The bioreactor system of claim 1, wherein the bioreactor has a volume greater than or equal to about 500 liters.

34. The bioreactor system of claim 1, wherein the bioreactor has a volume greater than or equal to about 1,000 liters.

35. The bioreactor system of claim 1, wherein the bioreactor has a volume greater than or equal to about 1,500 liters.

36. The bioreactor system of claim 1, wherein the bioreactor has a volume equal to between about 2,000 liters and about 3,500 liters.

37. The bioreactor system of claim 1, wherein the bioreactor has a volume equal to between about 2,000 liters and about 7,000 liters.

38. The bioreactor system of claim 1, wherein the bioreactor has at least two input ports and at least two output ports, and at least two of b), c), and d).

39. The bioreactor system of claim 1, wherein the bioreactor system is capable of operating in a perfusion process.

40. The bioreactor system of claim 1, wherein the bioreactor system is capable of operating during a harvesting process.

41. The bioreactor system of claim 1, wherein the bioreactor system is capable of operating during a perfusion process, an intermediate perfusion and harvest process, and a harvest process.

42. The bioreactor system of claim 1, wherein the bioreactor system is capable of operating in more than one perfusion process.

43. The bioreactor system of claim 1, wherein the bioreactor system is capable of operating in more than one perfusion process, intermediate perfusion and harvest process, and harvest process.

44. The bioreactor system of claim 1, wherein the bioreactor system is capable of operating in a continuous harvesting process.

45. The bioreactor system of claim 1, further comprising a human machine interface (HMI) control unit.

46. ​​The bioreactor system of claim 45, wherein the HMI control unit is programmed and can display perfusion process control, intermediate perfusion and harvest process control or harvest process control.

47. The bioreactor system of claim 1, further comprising one or more sensors.

48. The bioreactor system of claim 1, further comprising one or more of a feed stream fluid pump or a retentate stream fluid pump.

49. The bioreactor system of claim 1, further comprising at least one component capable of facilitating at least one process intensification parameter.

50. The bioreactor system of claim 49, wherein the process intensification parameter is one or more of increased cell number, increased cell density, provision of enriched cell culture growth medium, rapid cell number expansion, or increased biologic yield.

51. The bioreactor system of claim 1 further capable of backflushing the HF filter with a permeate fluid.

52. The bioreactor system of claim 1, 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 LPM to about 10 LPM or about 0.5 LPM to about 8 LPM.

53. The bioreactor system of claim 1, wherein the second pump is operable to pump the liquid at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 Fluid is drawn from the HF filter at a throughput of 100 Å to 200 Å.

54. The bioreactor system of claim 1, wherein the second pump is operable to pump the liquid at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 60,000 L / m 2 Between and / or around 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 Fluid is drawn from the HF filter at a throughput of 100 Å to 200 Å.

55. The bioreactor system of claim 1, wherein the second pump is operable to draw fluid from the HF filter at a filter flux rate of between about 150 liters per square meter per hour and about 900 liters per square meter per hour (LMH).

56. The bioreactor system of claim 1, wherein the second pump is operable to draw fluid from the HF filter at a filter flux rate of between about 200 LMH and about 700 LMH.

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

58. The bioreactor system of claim 1, wherein the second pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM and about 8 LPM.

59. The bioreactor system of claim 1, wherein the second pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM to about 13 LPM.

60. The bioreactor system of claim 1, wherein the second pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM to about 18 LPM.

61. The bioreactor system of claim 1, wherein the first flow path comprises a connection system having 3 / 8" ID tubing.

62. The bioreactor system of claim 1, wherein the first flow path comprises a connection system having 1 / 8" ID tubing.

63. The bioreactor system of claim 1, wherein the second flow path comprises a connection system having 1 / 2" ID tubing.

64. The bioreactor system of claim 1, wherein the second flow path comprises a connection system having 1 / 8" ID tubing.

65. The bioreactor system of claim 1, wherein the first flow path has an accuracy requirement of approximately 1%.

66. The bioreactor system of claim 1, wherein the second flow path has an accuracy requirement of approximately 3%.

67. The bioreactor system of claim 1, wherein the HF filter is constructed of and / or comprises polypropylene and / or polyethylene terephthalate.

68. The bioreactor system of claim 1, wherein the HF filter comprises: Isotropic pore structure; and / or pores having an average lumen diameter of about 0.65 µm to about 8 µm.

69. The bioreactor system of claim 1, operable for use with shear sensitive cells.

70. The bioreactor system of claim 1, operable for use with animal cells.

71. The bioreactor system of claim 1 , operable for use with mammalian cells.

72. The bioreactor system of claim 1, operable for use with Chinese Hamster Ovary (CHO) cells.

73. The bioreactor system of claim 1 , operable for use with human fetal kidney 293 (HEK293) cells.

74. The bioreactor system of claim 1 operable for use with cells to produce biologics.

75. The bioreactor system of claim 74, wherein the biologic comprises an antibody, a peptide and / or a virus.

76. A multi-purpose filter assembly comprising: a) Hollow fiber (HF) filters; as well as b) a multi-purpose assembly operably connected to the HF filter, the multi-purpose assembly comprising a first flow path and a second flow path: wherein the first flow path comprises a first pump operable to extract fluid from the HF filter and a flow meter operable to measure a flow rate of the fluid in the first flow path; wherein the second flow path comprises a second pump operable to draw fluid from the HF filter; and The first pump and the second pump can have different pump capacities and accuracy levels.

77. The multi-use filter assembly of claim 76, wherein the HF filter is operable for tangential flow.

78. A multi-purpose filter assembly according to claim 76, wherein the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter.

79. The multi-use filter assembly of claim 76, further comprising: One or more clarifying filters in the first flow path and / or the second flow path.

80. The multi-use filter assembly of claim 76, wherein the first pump and the second pump are selected from the group consisting of peristaltic pumps, centrifugal pumps, magnetic drive pumps, positive displacement pumps, membrane pumps, pressure-based pumps, quantex™ pumps, gear pumps, diaphragm pumps, syringe pumps, and piston pumps.

81. The multi-use filter assembly of claim 76, wherein the first pump and the second pump are peristaltic pumps.

82. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to pump the fluid at a rate of greater than or equal to about 10,000 liters per square meter (L / m 2 ) to about 30,000 L / m 2 The throughput of fluid is drawn from the HF filter.

83. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to pump the fluid at a rate of greater than or equal to about 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 The throughput of fluid is drawn from the HF filter.

84. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw a feed flow rate from the HF filter of about 0.8 liters per fiber per minute to about 2.2 liters per fiber per minute (L / fiber / min).

85. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw a feed flow rate of about 0.8 L / fiber / min to about 2.0 L / fiber / min from the HF filter.

86. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw a feed flow rate from the HF filter of about 1 L / fiber / min to about 1.8 L / fiber / min.

87. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a filter throughput of about 50 LMH to about 800 LMH.

88. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a filter throughput of about 100 LMH to about 600 LMH.

89. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to maintain a hematocrit (PCV) of about 2% to about 40%.

90. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to maintain a PCV of about 8% to about 40%.

91. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to maintain a PCV of about 12% to about 35%.

92. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter while maintaining a flow rate greater than about 25 x 10 6 cells / mL of VCD.

93. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter while maintaining a flow rate greater than about 50 x 10 6 cells / mL of VCD.

94. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter while maintaining a flow rate greater than about 75 x 10 6 cells / mL of VCD.

95. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter while maintaining a flow rate greater than about 100 x 10 6 cells / mL of VCD.

96. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter while maintaining a flow rate greater than about 25 x 10 7 cells / mL of VCD.

97. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a shear rate of less than about 5,000 s-1.

98. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a shear rate of less than about 3,500 s-1.

99. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a shear rate of less than about 2,500 s-1.

100. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM and about 5 LPM.

101. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM and about 10 LPM.

102. The multi-use filter assembly of claim 76, wherein the first flow path and pump are operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM and about 15 LPM.

103. The multi-use filter assembly of claim 76, wherein the flow meter operable to measure flow of fluid in the first flow path continuously monitors an LPM rate and communicates with the first pump to adjust the first pump rate to maintain a desired LPM rate.

104. The multi-use filter assembly of claim 76, wherein said flow meter operable to measure the flow rate of fluid in said first flow path is capable of accurately monitoring a flow rate of about 0 LPM to about 8 LPM.

105. The multi-use filter assembly of claim 76, wherein said flow meter operable to measure the flow rate of fluid in said first flow path is capable of accurately monitoring a flow rate of about 0.01 LPM to about 6 LPM.

106. A multi-purpose filter assembly according to claim 76, wherein the first flow path has an accuracy requirement of approximately 1%.

107. A multi-purpose filter assembly according to claim 76, wherein the second flow path has an accuracy requirement of approximately 3%.

108. The multi-use filter assembly of claim 76, wherein the first flow path and the second flow path have the same inner diameter or have different inner diameters.

109. The multi-use filter assembly of claim 76, wherein 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 first pump.

110. The multi-use filter assembly of claim 76, wherein the second flow path and pump are operable to extract fluid from the HF filter as a continuous harvesting process.

111. The multi-use filter assembly of claim 76, wherein the HF filter is not replaced while the multi-use assembly directs fluid through one flow path and pump and then directs fluid through another flow path and pump.

112. The multi-use filter assembly of claim 76, wherein the HF filter is not replaced when the multi-use assembly directs fluid first through the first flow path and pump and then through the second flow path and pump.

113. The multi-use filter assembly of claim 76, wherein the first flow path and the second flow path of the multi-use assembly are connected by a T-connector, a Y-connector, or a valve.

114. The multi-use filter assembly of claim 76, further comprising a second flow meter operable to measure the flow rate of fluid in the second flow path and accurately monitor a flow rate of about 0 LPM to about 10 LPM or about 0.5 LPM to about 8 LPM.

115. The multi-use filter assembly of claim 76, wherein the second pump is operable to pump at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 36,000 L / m 2 Fluid is drawn from the HF filter at a throughput of 100 Å to 200 Å.

116. The multi-use filter assembly of claim 76, wherein the second pump is operable to pump at a rate equal to about 12,000 liters per square meter (L / m 2 ) to about 60,000 L / m 2 Between and / or around 10,000 liters per square meter (L / m 2 ) to about 70,000 L / m 2 Fluid is drawn from the HF filter at a throughput of 100 Å to 200 Å.

117. The multi-use filter assembly of claim 76, wherein the second pump is operable to draw fluid from the HF filter at a filter flux rate of between about 150 liters per square meter per hour and about 900 liters per square meter per hour (LMH).

118. The multi-use filter assembly of claim 76, wherein the second pump is operable to draw fluid from the HF filter at a filter flux rate of between about 200 LMH and about 700 LMH.

119. The multi-use filter assembly of claim 76, wherein the second flow path and pump are operable to draw a feed flow rate from the HF filter of about 1 liter per fiber per minute to about 3 liters per fiber per minute (L / fiber / min).

120. The multi-use filter assembly of claim 76, wherein the second pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM and about 8 LPM.

121. The multi-use filter assembly of claim 76, wherein the second pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM and about 13 LPM.

122. The multi-use filter assembly of claim 76, wherein the second pump is operable to draw fluid from the HF filter at a flow rate between about 0.01 LPM and about 18 LPM.

123. The multi-use filter assembly of claim 76, wherein the HF filter is constructed from and / or comprises polypropylene and / or polyethylene terephthalate.

124. The multi-use filter assembly of claim 76, wherein the HF filter comprises: Isotropic pore structure; and / or pores having an average lumen diameter of about 0.65 µm to about 8 µm.

125. The multi-use filter assembly of claim 76, operable for use with shear sensitive cells.

126. The multi-use filter assembly of claim 76, operable for use with animal cells.

127. The multi-use filter assembly of claim 76, operable for use with mammalian cells.

128. The multi-use filter assembly of claim 76, operable for use with Chinese Hamster Ovary (CHO) cells.

129. The multi-use filter assembly of claim 76, operable for use with human fetal kidney 293 (HEK293) cells.

130. The multi-use filter assembly of claim 76 operable for use with cells to produce biologics.

131. The multi-use filter assembly of claim 130, wherein the biological agent comprises antibodies, peptides and / or viruses.

132. A method for producing a biologic from a cell, comprising: a) expanding cells in cell culture medium in a bioreactor, b) during cell expansion, perfusing cell culture fluid through a hollow fiber (HF) filter to remove spent cell culture medium while retaining expanding cells, and adding an appropriate replacement volume of cell culture medium to the bioreactor to maintain a desired cell culture fluid content in the bioreactor, wherein the spent cell culture medium is perfused through the HF filter at a first flow rate to enter a first flow path, and c) harvesting the cell culture fluid through the HF filter after cell expansion to obtain the biologic, and adding an appropriate replacement volume of cell culture medium to the bioreactor to maintain a desired cell culture fluid content in the bioreactor during different harvest stages, wherein the cell culture medium is harvested through the HF filter at a second flow rate to enter a second flow path, wherein the second flow rate is greater than the first flow rate.

133. The method of claim 132, wherein the HF filter is operable for tangential flow.

134. The method of claim 132, wherein the HF filter is a Tangential Flow Depth Filtration® (TFDF®) filter.

135. The method of claim 132, wherein the replacement volume of the cell culture medium in b) and / or c) is about the same as the amount of spent cell culture medium removed.

136. The method of claim 132, wherein the replacement volume of cell culture medium in b) and / or c) is greater than the amount of spent cell culture medium removed.

137. The method of claim 132, wherein the replacement volume of cell culture medium in b) and / or c) is less than the amount of spent cell culture medium removed.

138. The method of claim 132, wherein the replacement volume of cell culture medium is fresh cell culture medium.

139. The method of claim 132, wherein the perfusion process comprises more than one perfusion phase.

140. The method of claim 139, wherein the HF filter is used in more than one perfusion stage, an intermediate perfusion and harvesting process, and a harvesting process.

141. The method of claim 132, wherein the first flow rate is less than or equal to approximately 5 LPM and the second flow rate is greater than approximately 5 LPM.

142. The method according to claim 132, wherein the HF filter consists of and / or comprises polypropylene and / or polyethylene terephthalate.

143. The method of claim 132, wherein the HF filter comprises: an isotropic pore structure; and / or pores having an average lumen diameter of about 0.65 µm to about 8 µm.

144. The method of claim 132, wherein the cells are shear-sensitive cells.

145. The method of claim 132, wherein the cell is an animal cell.

146. The method of claim 132, wherein the cell is a mammalian cell.

147. The method of claim 132, wherein the cells are CHO cells.

148. The method of claim 132, wherein the cells are HEK293 cells.

149. The method of claim 132, wherein the biologic comprises an antibody, a peptide, and / or a virus.

150. A method of producing a biologic from cells comprising using the bioreactor system or multi-use filter assembly according to any one of claims 1 to 131.

151. Use of the multi-use filter assembly according to any one of claims 76 to 131 in a bioreactor system.

152. The use according to claim 151, wherein the bioreactor system comprises the features according to any one of claims 1 to 75.

153. Use of the bioreactor system according to any one of claims 1 to 76 for the cultivation of cells.

154. The use according to claim 153, wherein the cells produce a biologic and the use further comprises harvesting the cell culture fluid through the HF filter to obtain the biologic.

155. The system of any one of claims 1 to 75, wherein the system results in one or more of increased productivity, improved performance, increased efficiency, reduced contamination rates, reduced capital investment, reduced physical footprint, reduced consumable waste, and / or reduced operating costs for a facility comprising a biologics production process.

156. The component of any one of claims 76 to 131, wherein the component results in one or more of increased productivity, improved performance, increased efficiency, reduced contamination rates, reduced capital investment, reduced physical footprint, reduced consumable waste, and / or reduced operating costs for a facility comprising a biologic production process.

157. The method of any one of claims 132 to 150, wherein the method results in one or more of increased productivity, improved performance, increased efficiency, reduced contamination rates, reduced capital investment, reduced physical footprint, reduced consumables loss, and / or reduced operating costs for a facility comprising a biologics production process.

158. The use of any one of claims 151 to 154, wherein the use results in one or more of increased productivity, improved performance, increased efficiency, reduced contamination rates, reduced capital investment, reduced physical footprint, reduced consumables loss and / or reduced operating costs for a facility comprising a biologics production process.

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