Depth and virus filter (DF / VF) carts for batch and continuous processing

Abstract

A portable, automated, single-use filter cart system for continuous or batch replaceable biologics production processes is disclosed.

Description

[Technical field]

[0001] The present invention relates to an automated production facility, a production apparatus, and a production method for industrial production of a protein. [Background technology]

[0002] The biopharmaceutical industry is undergoing major change, driven in part by a rapid increase in approvals of novel biotherapeutics, improving protein expression rates, and increasing pressure from the biosimilar market (Non-Patent Document 1).

[0003] The rapid increase in the pharmaceutical market share of biological products (from 11% in 2002 to approximately 20% in 2017), combined with the need for access to affordable medicines in developing regions of the world, calls for the development of rapid, sustainable and cost-effective production methods (Non-Patent Document 2).

[0004] Biopharmaceutical production technologies that replace traditional batch processing platforms offer key advantages such as increased throughput, operational flexibility, and cost savings, as well as reduced environmental impact due to reduced footprint. Bioprocessing plants are designed to include continuous production processes with integrated upstream and downstream unit operations, allowing for rapid equipment changeover, product and capacity flexibility, and reduced production costs compared to batch culture processing (see, for example, Non-Patent Documents 3, 4, 5, and 6).

[0005] The advent of continuous perfusion technology has supported greater progress in connecting upstream process equipment to operate in continuous mode. This processing strategy has been valuable to several companies over the past 25 years and has helped them overcome product-related stability issues (Non-Patent Document 7).

[0006] Recent cell lines and media have been designed to target higher cell densities, especially in contrast to fed-batch processing, with some cultures achieving viable cell densities of over 100 million cells / mL (Non-Patent Document 8). As a result, the bottleneck of a typical biologics production facility has shifted from the production bioreactor (upstream process) to the purification train (downstream process), especially the chromatography column due to its dimensional limitations. Purifying the large batch sizes of product produced by the increasing amounts of protein derived from current production cell lines is no easy task (Non-Patent Document 9). Thus, the problem of integrating upstream biologics production processes with downstream processes continues to plague the biologics production industry.

[0007] Warikoo et al. reported that by integrating a continuous capture chromatography step downstream of the production bioreactor, a reduction in column size and buffer usage was achieved (Non-Patent Document 10).

[0008] have demonstrated that end-to-end continuous bioprocessing is feasible but still faces several challenges, including the development of robust viral clearance and automation strategies to ensure high product quality (11).

[0009] Vandiver et al. further developed a flexible biologics production technology capable of various production modes, including batch, fed-batch culture, enhanced fed-batch culture, semi-continuous or continuous end-to-end processes (Patent Document 1).

[0010] Previously known equipment and systems in the art, including filter carts, have been inadequate for modern biological production lines employing flexible configurations (see, for example, U.S. Patent Nos. 5,393,963, 5,393,132 and 5,443,636).

[0011] Thus, what is needed is a dual-pass depth and virus filter (DF / VF) cart capable of efficiently and flexibly meeting batch and / or continuous biologics (biopharmaceutical) manufacturing process requirements over long production periods. This is provided by the present invention. [Prior art documents] [Non-patent literature]

[0012] [Non-Patent Document 1] Levine et al., Efficient, flexible facilities for the 21st century, BioProcess International 10(11):20-30 (2012) [Non-Patent Document 2] Walsh, Biopharmaceutical benchmarks 2014, Nature biotechnology 32(10):992-1002 (2014) [Non-Patent Document 3] Farid et al., Evaluating the economic and operational feasibility of continuous processes for monoclonal antibodies, Continuous Processing in Pharmaceutical Manufacturing pp. 433-456 (2015) [Non-Patent Document 4] Kelley, Industrialization of mAb production technology: the bioprocessing industry at a crossroads, mAbs 1(5):443-452 (2009) [Non-Patent Document 5] Croughan et al., The future of industrial bioprocessing: Batch or continuous?, Biotechnology and Bioengineering 112:648-651 (2015) [Non-Patent Document 6] Pollock et al., Fed-batch and perfusion culture processes: Economic, environmental, and operational feasibility under uncertainty, Biotechnology and Bioengineering 110(1):206-219 (2013) [Non-Patent Document 7] Konstantinov et al., White paper on continuous bioprocessing, Journal of Pharmaceutical Sciences 104(3):813-820 (2015) [Non-Patent Document 8] Clincke et al., Very high density of chinese hamster ovary cells in perfusion by alternating tangential flow or tangential flow filtration in wave bioreactorTM-part ii: Applications for antibody production and cryopreservation, Biotechnology Progress 29(3):768-777 (2013) [Non-Patent Document 9] Chon et al., Advances in the production and downstream processing of antibodies, New Biotechnology 28(5):458-463 (2011) [Non-Patent Document 10] Warikoo et al., Integrated continuous production of recombinant therapeutic proteins, Biotechnology and Bioengineering 109(12):3018-3029.(2012) [Non-Patent Document 11] Godawat et al., End-to-end integrated fully continuous production of recombinant monoclonal antibodies, Journal of Biotechnology 213:13-19 (2015) [Patent documents]

[0013] [Patent Document 1] Vandiver et al., Automated Biological Production Systems, Equipment, and Processes, International Publication No. WO 2020 / 168315 [Patent Document 2] Lu et al., Perfusion Bioreactor with Filtration System, U.S. Patent Application Publication No. 2019 / 338238 [Patent Document 3] Born et al., Continuous Separator Bypass System and Method of Use Thereof, U.S. Patent Application Publication No. 2020 / 384380 [Patent Document 4] Reinbigler et al., Installation for Treating Biological Liquids, U.S. Patent Application Publication No. 2012 / 0248025 Summary of the Invention

[0014] The present invention includes a portable, automated, single-use filter cart system for the production of continuous or batch replaceable biologics.

[0015] The portable automated single-use filter cart system of the present invention comprises the following (a) and (b):

[0016] (a) A portable cart having an outer edge;

[0017] (b) a valved product inlet within the periphery of the portable cart for receiving an aqueous product-containing fluid from an upstream unit operation outside the periphery of the cart, wherein the upstream unit operation is fluidly connectable to the product inlet by a sealable connector and connecting lines, the product inlet being connected to a first pump or, interchangeably, to a second pump via connecting lines capable of fluidly transporting the aqueous product-containing fluid downstream, the first pump being adapted for low flow rate operation and the second pump being adapted for high flow rate operation, the first pump and the second pump each being configured to push a flow of the aqueous product-containing fluid through the first filtration path or, alternately, through the second filtration path, and wherein the first filtration path and the second filtration path each comprise: (i) a valved product inlet;

[0018] (i) a plurality of filters downstream of the first pump, the plurality of filters being fluidly connected in series along a filtration pathway; and, optionally, one or more portable accessory filtration module(s) outside an outer periphery of the cart, the one or more portable accessory filtration module(s) being fluidly connected to the filtration pathway and capable of receiving the aqueous product-containing fluid therefrom and returning filtered aqueous product-containing fluid thereto; (ii) a plurality of sensors capable of continuously measuring flow rate, flow rate, and / or pressure at one or more locations along the filtration path and transmitting measurement signals to a controller; and (iii) A connecting line capable of fluidly transporting the filtered aqueous product-containing fluid to a downstream product outlet port.

[0019] The portable, automated, single-use filter cart system of the present invention comprises the following (c) and (d):

[0020] (c) one or more valved buffer inlets within the periphery of the cart and upstream of the first and second filtration pathways, each buffer inlet fluidly connected by a connecting line capable of fluidly conveying a desired aqueous fluid received from a reservoir or an upstream unit operation for injection into the first or second filtration pathway by operation of the first or second pump and the valved buffer inlet in response to a command signal from the controller;

[0021] (d) the control device is configured and programmable to automatically switch the flow of product-containing fluid from the first filtration path to the second filtration path in response to a predetermined measurement signal received from a sensor(s) of the first filtration path or after a predetermined period of time, wherein the control device is further configured to automatically switch the flow of aqueous product-containing fluid from the second filtration path to the first filtration path in response to a predetermined measurement signal received from a sensor(s) of the second filtration path, and the control device is further configured to issue the following instruction signals (i) to (iii): (i) operating a first pump during a desired continuous biologics production process, or, interchangeably, operating a second pump during a desired batch biologics production process; (ii) operating the second pump during a desired continuous biological product production process to flow buffer or water through the first filtration path while the flow of aqueous product-containing fluid passes through the second filtration path, or, interchangeably, to flow buffer or water through the second filtration path while the flow of aqueous product-containing fluid passes through the first filtration path; and (iii) controlling a valved product inlet and one or more valved buffer inlets;

[0022] One advantage of the present invention is that during a desired continuous biological product production process, the portable, automated, single-use DF / VF filter cart of the present invention supports switching between a first and second filtration pathway an unlimited number of times as required by the execution of the process, even if some manual intervention is required to change filters as needed.

[0023] The foregoing summary is not intended to define all aspects of the invention, and additional aspects are described in other sections, such as the "Detailed Description" of the embodiments. It should be understood that the entire specification is intended to be linked as a unified disclosure, and that all combinations of features described herein are contemplated, even if the combinations of features are not found together in the same sentence, paragraph, or section of the specification. For example, it should be understood that certain aspects of the invention described as a genus, and all members of the genus individually, are aspects of the invention. [Brief description of the drawings]

[0024] [Figure 1] FIG. 1 illustrates an embodiment of a portable automated single-use filter cart system. [Diagram 2] FIG. 2 shows the embodiment of FIG. 1 from a different perspective. [Diagram 3] 3 shows another embodiment including an "off-skid" filtration module. This embodiment of a portable automated single-use filter cart is shown with depth filters (326a) housed in housings or holders on the outer periphery of the cart in two portable accessory filtration modules, one for each of the first and second filtration paths. [Figure 4] FIG. 4 shows a schematic diagram of a portable, automated, single-use DFVF filter cart system design of the present invention capable of operating either depth filters (426a) or virus filters (426b) in either batch or continuous mode. [Diagram 5]FIG. 5 shows the load inlet pressure and load turbidity over time for a depth filter in a representative run using a depth filter as described in Example 1 herein. [Figure 6] FIG. 6 shows the virus filter loading flow rate and differential pressure over time in a representative run using a virus filter as described in Example 2 herein. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0026] [Definition] Unless otherwise defined herein, scientific and technical terms used in connection with this application shall have the meanings commonly understood by those skilled in the art. Furthermore, unless the context dictates otherwise, singular terms shall include the plural and plural terms shall include the singular. Thus, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, a reference to "a protein" includes a plurality of proteins, and a reference to "a cell" includes a collection of a plurality of cells.

[0027] The present invention encompasses a portable, automated, single-use filter cart system for interchangeable continuous or batch biological product production. The filter cart of the present invention can be performed under aseptic operating conditions and can be incorporated into an automatically controlled and regulated biological product production process as needed, regardless of whether the production process is a batch process, semi-continuous process, or continuous process. The filter cart system of the present invention can function interchangeably in any of these processing modes and can be placed between different unit operations as needed for a particular production process.

[0028] The term "integrated" in the context of a process for producing a purified protein or other biological product of interest (e.g., but not limited to, a protein pharmaceutical agent) means that one or more upstream and / or downstream steps in the production process are performed under common or coordinated control based on programmed commands that are modified by current sensor feedback of parameters defined in terms of set points or flow rates. The term "coordinated" means that two or more operations, processes, steps, components, or systems are controlled, regulated, or scheduled in such a relationship that ensures efficiency or harmony of their functioning toward a single purpose.

[0029] In a typical biologics production process, the cell culture-containing fluid containing the protein or other biologic of interest (e.g., an antibody) is harvested, after which the product can be further purified from the cell-free supernatant fraction. Typically, protein purification is usually achieved by a series of any of a number of chromatographic steps, such as anion exchange chromatography, cation exchange chromatography, affinity chromatography (using Protein A or Protein G or Protein L as affinity ligands), hydrophobic interaction chromatography, hydroxyapatite chromatography, and size exclusion chromatography. In addition, the purification step may include one or more depth filtration, virus filtration, ultrafiltration, nanofiltration, or diafiltration steps, and / or, optionally, an acid virus inactivation step. Depending on the biologic to be recovered, any other known technique for protein purification, such as ethanol precipitation, reversed-phase HPLC, chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation, is also possible.

[0030] In general, "upstream" processing steps include, but are not limited to, for example, culturing product producing cells (e.g., but not limited to recombinant host cells) in a bioreactor (e.g., perfusion bioreactor); removing cells from the permeate; and fluidly feeding a large volume of cell-free permeate from the bioreactor(s), optionally into an intervening surge vessel or directly to a downstream processing step. "Downstream" processing steps can include, but are not limited to, for example, capture and purification of the product in at least one or more chromatography systems; a virus inactivation system, such as a low pH virus inactivation system or a detergent virus inactivation system, and optionally (e.g., in embodiments of a low pH virus inactivation system), a neutralization system; a depth filtration system; a virus filtration system; and / or an ultrafiltration / diafiltration system. In a given production run, the order or details of these processing steps may vary. However, a first production processing step or unit operation can also be said to be "upstream" with respect to a second unit operation that follows (directly or indirectly) the first unit operation in a production run, and a second unit operation can also be said to be "downstream" with respect to a first unit operation that precedes (directly or indirectly) the second unit operation. Within a unit operation, for example, within the portable automated single-use filter cart system of the invention, "upstream" refers to a first location or function that precedes a second location or function in the system (based on the direction of flow of product-containing fluid through the system), and the second location or function in the system is referred to as being "downstream" from the first location or function.

[0031] A "continuous" form or mode of production process or system refers to a mode of processing in which a perfusion bioreactor is fluidly connected (directly or indirectly through intervening unit operations) to a continuous capture chromatography step (e.g., processing with a first chromatography system) in an uninterrupted flow from the bioreactor to a first chromatography system, which is then fluidly connected in an uninterrupted flow to a downstream viral inactivation step, and optionally fluidly connected in an uninterrupted flow to a depth filter. Further downstream product purification steps (e.g., a second chromatography system, an optional third chromatography system, a viral filter, and processing with ultrafiltration / diafiltration) are fluidly connected in series with each other, with optional intervening surge receptacles, to the above upstream processing steps, all in an uninterrupted flow.

[0032] A "semi-continuous" format or mode of production process refers to a processing regime in which a perfusion bioreactor is fluidly connected in an uninterrupted flow to a continuous capture chromatography step (e.g., processing with a first chromatography system), and then fluidly connected to processing with another step, such as, but not limited to, a viral inactivation system, and optionally fluidly connected in an uninterrupted flow to a depth filter, storing the virally inactivated product pool in a holding vessel prior to further processing. The temporary storage of the product pool in the holding vessel is then followed by one or more batch downstream processing step(s), which step(s) may be fluidly connected to each other in an uninterrupted flow, optionally with an intervening surge or holding vessel (i.e., a holding vessel if there is more than one batch step or operation), in succession, for example, to a second chromatography system, an optional third chromatography system, and processing with ultrafiltration / diafiltration.

[0033] A "perfusion bioreactor" is a bioreactor for cell culture in which an equivalent amount of medium can be added to and removed from the reactor while the cells remain in the bioreactor. A perfusion bioreactor includes a bioreactor and an operably attached perfusion system. The perfusion system provides a steady source of fresh nutrient medium and removal of cellular waste products. Typically, the bioreactor and the perfusion system of a perfusion bioreactor may be separate mechanical units that work in concert. Many commercially available examples include, but are not limited to, Xcellerex™ brand single-use bioreactors (SUBs from GE Healthcare Life Sciences) and KrosFlo™ brand perfusion flowpath assemblies and systems (Spectrum from Repligen), and a variety of other bioreactors and perfusion systems that can be appropriately combined into one perfusion bioreactor by one skilled in the art. Alternatively, the bioreactor and the perfusion system can be assembled into a single mechanical unit, for example, but not limited to, a 3D Biotek brand perfusion bioreactor (Sigma-Aldrich). The protein product secreted in the bioreactor can be continuously harvested by microfiltration during the process of removing the medium through the perfusion system, and the protein of interest is thus isolated in the microfiltration filtrate exiting the perfusion system.

[0034] A step in a production process or a system in an automated production facility is "fluidically" performed or "fluidically connected" to or "fluidically" receives material from another step in the production process or another system when material containing the protein of interest flows between the steps or systems by pipes, tubes, or other closed conduits without manual input or output. Such pipes, tubes, or other closed conduits between steps or systems are considered "connecting lines" when they are capable of fluidly transporting aqueous product-containing fluids downstream within the system or between systems or unit operations. The connecting lines can be made of any suitable non-reactive, non-porous material, such as, but not limited to, silicone rubber, Teflon, stainless steel, and the like. Sterile connectors are useful for joining connecting lines to other components, as known to those skilled in the art.

[0035] A step in a production process or a system within an automated production facility is commonly referred to as a "unit operation." A "production line" or "processing line" (used interchangeably herein) is a series of unit operations involved in the production of a biological product, such as, but not limited to, a protein of interest, a pharmaceutical substance, or a drug product.

[0036] A "batch unit" or "skid" is a unit operation. Typical examples include a single-use bioreactor, a perfusion system, a first chromatography system, an optional second chromatography system, an optional third chromatography system, and / or an ultrafiltration / diafiltration system, each of which may be configured as a "skid", but need not be. A viral inactivation system, and optionally a neutralization system, may also be a "skid". A batch unit is configured to communicate (e.g., by hard wiring, fiber optic cable, or wireless connection) with a Process Control System (PCS) or a Supervisory Control and Data Acquisition (SCADA) system. The portable automated single-use filter cart system of the present invention may also be configured as a "skid" within a production process. A "non-batch unit" is a device that supports one or more batch units. A non-batch unit is configured to communicate (e.g., by hard wiring, fiber optic cable, or wireless connection) with a PCS or SCADA. Additionally, dongles and / or Profibus devices, or similar digital information storage devices and electronic hardware connector(s) can be attached to non-batch units to identify their location (relative to a batch unit) and role (e.g., supply tank, collection tank, filter bank, etc.). For convenience and flexibility, filter banks, heat exchangers, surge receptacles, supply tanks, reservoirs, holding vessels, collection or collection tanks (e.g., elution collection vessels), and, optionally, portable mixers and other mixing vessels, if present, are typically configured as non-batch units, although in some embodiments unit operations such as these may also be included on "skids" that include control via hard wiring, fiber optic cables, or wireless connections.

[0037] The terms "automated," "automatically controlled," or "automatically" are used interchangeably in reference to a production process or production facility and refer to computerized control of the implementation or execution of one or more process steps or the operation of a component or system of the production facility, optionally with feedback control of the process step or operation. Typically, the computerized controller receives digital signals from a detector of the physical or chemical parameter to be controlled and issues responsive digital instructions to a system or subsystem.

[0038] The terms "single-use" or "single-use" as used interchangeably herein mean that a particular sterile production line component, i.e., a piece of sterile equipment, such as a component used in the portable automated single-use filter cart system of the present invention, is constructed or configured to be used for one production run (although it can be reused if quality and aseptic hygiene can be guaranteed for multiple production runs). The single-use component can then be discarded and replaced with another single-use component of the same or modified configuration for a subsequent production run without the need for cleaning (flushing) and disinfecting the component between production runs. Such single-use components can be commercially constructed and obtained. Examples of single-use components that can be used in the present invention include, but are not limited to, depth filters, virus filters, pre-filters, sterilizing grade filters, or any of the following: connectors, connecting lines or valves, containers or bags. Single-use depth filters, single-use virus filters, single-use prefilters, and single-use sterilizing grade filters are widely available commercially and include, for example, single-use filter assembly systems including filters (various membranes and pore sizes available from MilliporeSigma or Sartorius Stedim Biotech); single-use virus filtration systems such as the Allegro™ MVP Single-Use System Manifold (Pall Biotech); Mobius™ FlexReady for virus filtration (MilliporeSigma); FlexAct™ for virus filtration (Sartorius), Planova™ Single-Use Virus Filters (SU-VFS; Asahi Kasei Bioprocess America), or Viresolve™ Pro Virus Filters (MilliporeSigma).Single-use connecting lines in various dimensions, lengths, and configurations using single-use silicone and / or c-flex type tubing are commercially available from Thermo Fisher Scientific (ASI), AdvantaPure, Pall, Colder, GE Healthcare Life Sciences, and Sartorius Stedim Biotech; systems of connecting lines are also known as "tubing manifolds." Single-use sterile connectors are available from a variety of manufacturers, including AseptiQuik™ connectors (Colder Products Company), Kleenpak™ pre-sterilized connectors (Pall Biotech); Lynx™ ST connectors (MilliporeSigma).

[0039] The terms "switching" or "switch," as used interchangeably herein with respect to a first filtration path and a second filtration path, refer to changing the direction of flow of an aqueous product-containing fluid from a first filtration path to a second filtration path, or vice versa. Such switching can be under the automated control and coordination of a computer, i.e., controller (140, 240, 440) and mechanism(s), such as valves (110, 210, 310, 410) and pumps (116a, 116b, 216a, 316a, 316b, 416a, 416b), and can optionally be controlled by an external control system, such as an automated production facility control system. Although the switching can be manually controlled and adjusted, "automatic switching" means that the switching of flow direction does not require manual input on the part of an operator, but instead is controlled by a controller (140, 240, 440), which performs the switching of flow direction under predetermined criteria or set points.

[0040] An "auxiliary filtration module" is a filtration unit located outside the outer periphery of the filter cart that is fluidly connected to and can be incorporated into either the first filtration path or the second filtration path (see, for example, the embodiment in Figure 3).

[0041] The terms "sterile" or "aseptic" as used herein refer to a material that is free or essentially free of microbial and / or viral contamination. In this regard, "contaminants" refers to materials that are different from the desired components in a preparation, such as a cell culture medium, a component of a cell culture medium, or an aqueous product-containing fluid, or any other preparation. In the context of "sterile filtration," the term sterile filtration is a functional description of filtering a preparation through a sterile filter (pore size 0.2 μm or less) to remove bacterial and / or mycoplasma contaminants.

[0042] "Batch filtration" or alternatively "batch filtration" or filtration performed in a batch mode, as used herein, refers to a process in which a certain amount or volume of a preparation, such as cell culture medium or at least a component of a cell culture medium, or an aqueous product-containing fluid, or any other preparation, is filtered in one batch through a filter, such as, but not limited to, a depth filter or a virus filter, depending on the filter capacity, and the filtration process is finalized before the filtrate is directed or fed to a process where it is used or consumed.

[0043] The terms "continuous filtration" or "on-line filtration" or "in-line filtration" refer to a filtration process in which a certain amount or volume of a preparation, be it cell culture medium or at least a component of a cell culture medium, or an aqueous product-containing fluid, or any other preparation, is continuously filtered through a filter, such as, but not limited to, a depth filter or a virus filter, depending on the capacity of the filter, and the filtration process is still ongoing as the filtrate is directed or fed to a process where it is already used or consumed.

[0044] "Cell culture supernatant" is the extracellular medium in which product-producing cells are cultured. This medium should not be confused with feed medium, which may be added to the culture after the cells are inoculated into the cell culture medium to initiate cell growth. "Cell culture" refers to the cell culture supernatant and the product-producing cells cultured therein. Typically, mammalian production cells are cultured at 37±1° C.

[0045] "Incubating" or "maintaining" at a given temperature or other condition parameter (e.g., pH, oxygenation, CO2 concentration, pressure, etc.) means that the process control system is set to the intended target temperature within the unit operation. The portable automated single-use filter cart system of the present invention can include such automatic adjustments to maintain the desired conditions. Incubation or unit operation conditions such as temperature (typically, but not necessarily, about 37°C), pH, oxygenation, etc. are maintained at the desired set points by digital control units (DCUs) and sensory monitors, which are commercially available or can be constructed by one of skill in the art. Digital control units (DCUs) that can control and monitor the state of unit operations within a production process are manufactured and commercially available by companies such as BBraun, New Brunswick, or Sartorius.

[0046] For any given condition set point, slight variations may occur during operation of the process. Suitable monitoring equipment and suitable alternatives are commercially available or can be constructed by one skilled in the art.

[0047] The term "buffer" or "buffer solution" refers to a solution that resists changes in pH by the action of its conjugate acid-base range. Examples of useful buffers that control pH in the range of about pH 4 to about pH 8 include phosphate, bicarbonate, acetate, MES, citrate, Tris, Bis-Tris, histidine, arginine, succinate, citrate, glutamate, and lactate, or combinations of two or more of these, or other mineral or organic acid buffers. Salts containing sodium, ammonium, and potassium cations are often used in preparing buffer solutions.

[0048] The term "loading buffer" or "equilibration buffer" refers to a buffer and salt or salts that are mixed with a product-containing preparation (e.g., supernatant or filtrate of a batch or perfusion culture, or an eluate pool containing the biological product of interest, e.g., an antibody) to load the protein preparation onto an affinity purification matrix, or, in some cases, onto a depth or viral filter. Affinity purification matrices include, for example, Protein A or Protein g conjugate matrices, or specific target ligand conjugate affinity chromatography matrices. The buffer is also used to equilibrate the applicable matrix or filter prior to loading and for washing after loading the biological product.

[0049] [cart] The portable automated single-use filter cart system of the present invention has a perimeter or boundary and includes a frame, a top, a base, and preferably wheels, spheres, sleds, casters, or other suitable members at the base to enhance portability of the cart (see, for example, (130) in FIG. 1, (230) in FIG. 2, and (330a) in FIG. 3). Wheels, spheres, sleds, casters, or other suitable members to enable or enhance portability can also be included at the base of the accessory filtration module (see, for example, (330b) in FIG. 3). The cart frame, cart top, base, and optional shelves, trays, drawers, hooks, clips, snaps, tubing included with the cart are considered to be located with the perimeter of the cart. In different embodiments, the shape of the cart can be modified to suit the desired application or purpose within the production line, but a generally rectangular or cubic three-dimensional shape is often most convenient for many applications (see, for example, FIG. 1, FIG. 2, and FIG. 3). The cart can be constructed of any suitable material, such as, but not limited to, stainless steel, aluminum, titanium, metal alloys, plastic, or any other suitable strong, durable, washable and / or sanitizable material, or combination of these materials. In one embodiment, 316L stainless steel is used to construct the cart due to the corrosion resistance of this material and its cleanability. The size of the cart can be varied as needed for a particular purpose. In some exemplary embodiments, the outer dimensions of the cart are approximately 65 inches by 32 inches by 45 inches (length x width x height).

[0050] In some embodiments, the controller is located within the perimeter of the cart, e.g., within the confines of the frame (see, e.g., 140 in FIG. 1 and 240 in FIG. 2), or on top of the cart. In other embodiments, the controller may be outside the perimeter of the cart itself, so long as the controller can communicate with the pumps, valves, and sensors on the filter cart, i.e., the controller can receive measurement signals transmitted from the sensors and the controller can send instructions to all of the pumps and valves. Such transmission and / or reception of measurement or instruction signals may be via electrical cables, wiring, fiber optic cables, as the case may be, or may be via infrared or radio wave transmission, and / or any other suitable medium.

[0051] In some embodiments, hooks (e.g., (170) in FIG. 1 and (270) in FIG. 2), loops, clips, and / or trays are included at any convenient location on the cart. Such hooks, loops, and / or clips are for cable management of power cords, signal cables, or wiring, and / or for management or support of tubing or plumbing. For example, trays or hooks to hold tubing or cables or wiring that extend beyond the periphery of the system can improve and maintain organization. In some embodiments, several small plastic clips are attached to the front and back of the cart (or other vertical cart portion) to hold the tubing in place.

[0052] [pump] The pumps (116a, 116b, 216a, 316b, 416a, 416b) can be peristaltic or positive displacement. The pump heads can be single-use product-contacting pump heads that can be replaced between runs, or can be reusable conventional product-non-contacting pump heads such as, but not limited to, a typical peristaltic pump. For example, in some embodiments, the second pump (for high flow operation) is a single-use quadruple diaphragm pump, Quattroflow 1200SU HT, with a flow range of about 6 to 1200 L / hr, and the first pump (for low flow operation) is a Quattroflow™ 150SU pump motor retrofitted with a Quattroflow™ 30SU pump head, with a flow range of about 0.06 to about 30 L / hr. Other pumps capable of operating in the aforementioned flow ranges are also useful in the DF / VF filter carts of the present invention, and many are commercially available.

[0053] [valve] The automatic valves of the portable automated single-use filter cart system of the present invention can be of any suitable type for regulating the flow of fluids through the connecting lines. In some embodiments, the automatic valves are robotically controlled mechanical valves, pneumatic valves, hydraulic valves, diaphragm valves, ball valves, gate valves, needle valves, bellows valves, or globe valves. Pneumatic valves are particularly useful when a source of compressed air is available. Pinch valve types, which have a block that drops onto the tubing of the connecting lines to pinch it closed, are useful embodiments (e.g., pinch valves manufactured by Aquasyn LLC or Acro Associates, a wholly owned subsidiary of Bimba Manufacturing Corporation). A variety of pinch valves are useful, including, but not limited to, overmolded tube assemblies to reduce dead volume in the tubing. Other useful options include multiplexing of the valves, such as, for example, 3-way or 4-way configurations. One of ordinary skill in the art knows how to select the most suitable valve for a given embodiment, taking into account the desired level of simplicity, cost-effectiveness, and the nature of the tube assembly selected in terms of minimizing the valve footprint and dead volume.

[0054] [Control devices and automated production processes] The controller (140, 240, 440) of the portable automated single-use filter cart system of the present invention can be configured and programmed to independently and automatically switch filtration paths of the filter cart system based on pre-programmed instructions, set points, and measurement signals sent from the cart's own pressure sensors (118, 218, 318, 418), flow meters (114, 314, 414), and other analytical sensors (120, 220, 320, 420) monitoring UV (420a), conductivity and temperature (420b, 420c), and pH (420d). The controller (140, 240, 440) can be configured and programmed to automatically switch the flow of the aqueous product-containing fluid from the first filtration path to the second filtration path, or vice versa, from the second filtration path to the first filtration path, in response to a predetermined measurement signal received from the sensor(s) of the second filtration path, and the controller can be further configured and programmed to issue the following instruction signals: (i) operate the first pump during a continuous production process of the desired biological product, or, interchangeably, operate the second pump during a batch production process of the desired biological product; (ii) operate the second pump during a continuous production process of the desired biological product to flow buffer or water through the first filtration path while the flow of the aqueous product-containing fluid is passing through the second filtration path, or, interchangeably, to flow buffer or water through the second filtration path while the flow of the aqueous product-containing fluid is passing through the first filtration path; and (iii) control the valved product inlet and the one or more valved buffer inlets.

[0055] However, in some embodiments, the controller (140, 240, 440) may also be configured and programmed to interact with a comprehensive production facility control system. In general, the production facility control system may be used to control an automated production line that includes individual unit operations and has a flexible configuration, such as the portable automated single-use filter cart system of the present invention. That is, the production facility control system may be designed to accommodate multiple configurations that utilize portable equipment that can be coupled to other components of the production line. The production line may include one or more skids that include original manufacturer equipment, such as a single-use bioreactor system, a perfusion system, or a continuous chromatography system. The skid may also include flow control devices, such as pumps. In addition, the skid may include one or more communication interfaces that allow the portable piece of equipment to be physically coupled to the skid. The physical coupling between the portable piece of equipment and the skid may be accomplished using electrical cables. The electrical cables may be configured to accommodate serial, bus, or Ethernet communications. In some embodiments, the electrical wiring may be Recommended Standard 232 (RS-232) wiring. It can be said that:

[0056] Portable pieces of equipment, such as the portable automated single-use filter cart system of the present invention, can include or be otherwise coupled to a network gateway hardware device that enables communication between the skid and a production facility control system. Additionally, at least some of the skids can be logically configured to be coupled to various portable pieces of equipment. In this manner, a piece of portable equipment can be physically connected to a particular skid based on the configuration of a particular production line, and the skid can be configured to operate in multiple configurations based on the different pieces of equipment coupled to the skid.

[0057] The location of the portable automated single-use filter cart system of the present invention for depth or virus filter operations in a production line can be assigned and / or identified by a production facility control system. Additionally, the specific sequence logic for a particular depth or virus filter process can also be facilitated by the production facility control system in the form of an automated recipe or method that complies with ISA-88 and ISA-95 (and / or other applicable) standards to control one or more control modules, flags, and / or status identifiers for the depth and / or virus filters and to execute one or more control modules simultaneously with the filter carts. The production facility control system can monitor various process conditions and process signals, such as pressure or flow rate.

[0058] For example, the pressure in the filter assemblies of the filter cart can be monitored based on pressure values ​​obtained from pressure sensors (118, 218, 318, 418) included in the filter cart assemblies and relayed by the filter cart controller (140, 240, 440). The production facility control system can determine that the pressure in the first filter assembly through which the material flows has reached at least a threshold level. The threshold level of pressure can indicate that the filter included in the first assembly needs to be replaced due to a decrease in the amount of material that can be processed by the filter, and can generate an indication signal and / or an alarm signal. The production facility control system can then send a signal to the controller (140, 240, 440) to switch the flow of the aqueous product-containing fluid from the first filtration path to the second filtration path by operation of a valve, as explained in more detail in the schematic diagram of FIG. 4. If necessary, for example, over a long production run, the filters included in the first and / or second filtration paths can be replaced. Similarly, the controller can be programmed to respond to other pre-set parameters (e.g., duration, volumetric throughput) or trigger conditions measured by analytical sensors (120, 220, 320, 420) that prompt the controller to switch the flow of aqueous product-containing fluid from one flow path to the other and / or respond to an operator prompt (i.e., manual intervention).

[0059] System Design and Exemplary Embodiments FIG. 4 shows a schematic diagram of a portable automated single-use DFVF filter cart system design of the present invention that can operate in either batch or continuous mode with either a depth filter (426a) in a so-called "stage 1" mode or position, or a virus filter (426b) in a so-called "stage 2" mode or position. As further shown in FIG. 4, the system design can accommodate various pre-filter, sterilizing grade filter (426c), and / or depth filter (426a), and / or virus filter (426b) configurations located on the filter cart (e.g., see 126b in FIG. 1) or, optionally, off the outer edge of the filter cart (shown as "P" in FIG. 4). These filters can be located in three different positions, also referred to as "stages," on the first or second filtration path. "Stage 1" refers to the first location in the filter path (426a), "Stage 2" refers to the second location in the filter path (426b), and "Stage 3" refers to the third location in the filter path. For example, in some embodiments, the depth filtration process can have a depth filter (426a) located in stage 1, located outside the outer periphery ("P") of the filter cart system (see also FIG. 3), and optionally a sterilizing grade filter (426c) located in stage 3 (i.e., within "P"; see e.g., 126c in FIG. 1) on the filter cart as shown diagrammatically in 426c. In this example, stage 2 location (426b) is bypassed by using a piece of tubing to connect the remainder of the tubing manifold section in lieu of a filter completing the flow path. In an exemplary virus filtering process, a pre-filter can be placed in stage 1 on the filter cart (i.e., within the cart perimeter "P") as shown by 426a, a virus filter (426b) can be placed in stage 2 either on the filter cart (i.e., within "P") or outside the perimeter (426b), and a sterilizing grade filter can be placed in stage 3 on the filter cart system (426c; within "P").The filter can be located in either the first filtration path or the second filtration path, or both, for a continuous production process with filter replacement.

[0060] The system also comprises a set of inlets (401, 402) including (i) one or more valved buffer inlets (402) and (ii) a product inlet (401) for receiving an aqueous product-containing fluid from an upstream unit operation fluidly connected thereto upstream of the first and second filtration paths. The inlets (401, 402) are fluidly connected via tubing to a set of inlet valves (410a, 410b) and a set of pumps (416a, 416b) for performing buffer flushing or product filtration operations.

[0061] The first pump (416a) is designed for low flow rate operation in a continuous production process and is used to load product-containing fluid from the product inlet (401) through the product inlet valve (410a) onto the filter during continuous processing. The second pump (416b) is designed for high flow rate operation and is used to perform buffer flushing, equilibration, or sanitization operations for either batch or continuous processing, the second pump (416b) is also used to load product onto the filters (426a, 426b, 426c) during batch processing. The system also includes a set of inlets (401, 402) including (i) one or more valved buffer inlets (402) for receiving aqueous product-containing fluid from an upstream unit operation fluidly connected thereto upstream of the first and second filtration paths, and (ii) a product inlet (401). The product-containing fluid is fluidly delivered to the product inlet (401) via tubing, or the buffer(s) to be used are fluidly delivered to one or more buffer inlets (402) via tubing. The inlets (401, 402) are also fluidly connected via tubing to a set of inlet valves (410a, 410b) and a set of pumps (416a, 416b) to perform buffer flushing or product filtration operations. The selection of the inlet or inlets (402) and which buffer inlet (402) is operated is automatically controlled by the buffer inlet valve (410b), and the automation of controlling the product inlet valve (410a) and the buffer inlet valve (410b) is under the direction of a controller (440). In the schematic diagram of FIG. 4, the controller (440) is shown as being outside the periphery ("P") of the cart, however, this is merely for convenience and clarity of drawing; the controller could be inside the periphery of the cart (e.g., see FIG. 1 (140) and FIG. 2 (240)), or, in some cases, outside the periphery of the cart, so long as measurement and / or indication signals can be easily exchanged between the analytical sensors (418, 420), valves (410), pumps (416a, 416b), and controller (440).The pumps (416a, 416b) can be operated in either (i) a flow control mode using flow meters (414) configured in both the first and second filtration paths, or (ii) a pressure control mode using one or more pressure sensors (418) configured at intervals in both the first and second filtration paths. For example, in FIG. 4, pressure sensors (418) are positioned in both the first and second filtration paths before the first filter position (i.e., before the DF or "stage 1"; 426a), between the first (DF) position (426a) and the second filter (VF; 426b) position (i.e., "stage 2"), and between the second filter (VF; 426b) and the third filter (i.e., "stage 3" or sterilizing grade filter; 426c).

[0062] In flow control mode, the automation adjusts the pump output (416a or 416b) based on a feedback loop from measurements on the corresponding flow meter (414) to achieve a preset and configured set point in the automation. In pressure control mode, the automation adjusts the pump output (416a or 416b) based on a feedback loop from measurements on the pressure sensor (418). In this example, the pressure differential across the virus filter (426b) is used to achieve a preset and configured set point in the automation using pressure sensors placed before and after the second filter position.

[0063] For batch production processes, Figure 4 shows "crossover" valves (410c, 410d, 410e, 410f) that can be automatically positioned to direct product or buffer flow from the high flow pump (416b) to the first filtration path (i.e., 410e open, 410f and 410c closed) or to the second filtration path (i.e., 410f open, 410d and 410e closed). For continuous production processes, the crossover valves can be automatically positioned to direct product from the low flow pump (416a) to the first filtration path (i.e., 410c open, 410d and 410e closed) or to the second filtration path (i.e., 410c and 410f closed, 410d open). For buffer flushing in a continuous process, the crossover valve can be automatically positioned to direct buffer flow from the high flow pump (416b) to the first filtration path (i.e., 410e open, 410f and 410c closed) or to the second filtration path (i.e., 410f open, 410d and 410e closed). In a continuous process, buffer flushing via the high flow pump (416b) can occur simultaneously with product filtration via the low flow pump (416a).

[0064] During the buffer flushing operation, the waste outlet valves (410g) configured on both the first and second filtration paths can be used to flush (bulk) buffer for the stage 1 or stage 2 filters. While product filtration is initiated, the residual volume in the first and second filtration paths can be flushed to waste by flowing through valves (410i), (410g), and / or (410j) to the waste ports (waste ports are not shown in FIG. 4, but see, for example, (104) in FIG. 1 and (204) in FIG. 2). When the product is switched to product collection through the product outlet ports (103, 203, 403) toward the downstream unit operation, one of the product outlet valves (410k) can be released to the product outlet port (403). The manual valve (412) is used to introduce air into the virus filter (426b) for pressure hold or air diffusion integrity testing.

[0065] Analytical sensors (420a, 420b, 420c, 420d) are placed at or near the product outlet port (403) of the filter cart system to monitor UV (420a), conductivity (420b, 420c), and pH (420d). The sensors can be used in automation to trigger the start or end of buffer flushes or product harvests.

[0066] 1 and 2 show an embodiment of a portable automated single-use filter cart system from different perspectives. Portability can be optionally enhanced by wheels (130, 230) and handles (150, 250) for manipulating the position of the filter cart. One or more other optional buffer inlets (shown as 102 in FIG. 1 but not shown in the perspective view of FIG. 2) are present on the filter cart to introduce buffer(s) or water into the first or second filtration path for buffer flushing or product filtration operations. In this embodiment, each of the first and second filtration paths includes a pre-filter (e.g., 126a, 226a), or a depth filter, or a viral filter (e.g., 126b, 226b), and / or a sterilizing grade filter (126c, 226c). In the perspective of Figures 1 and 2, a first filtration path is shown at the top of the cart (within the outer periphery of the cart) as an upper path further from the viewer, and a second filtration path is shown as a lower path closer to the viewer. The system employs automated valves (110, 210) controlled by a controller (140, 240) to direct the flow of aqueous product-containing fluid conveyed through a connecting line of tubing. That is, the initial flow enters the system from an upstream unit operation through an automated product inlet valve (110a) via a product inlet (101, 201), while optionally, one or more buffer inlets (102; not shown in the perspective view of Figure 2) convey buffer or water into the system via an automated buffer inlet valve (110b; not shown in the perspective view of Figure 2). The automated valves (110, 210) in this embodiment and many other embodiments of the invention are pneumatic valves and are robotically actuated by compressed air conveyed by a separate air piping system in the cart. Any suitable compressed air source can be employed. 1 and 2 show air intakes (161, 261) for a compressed air piping system, and the perspective view of FIG. 2 shows an air regulator (260) for the air piping system.Manual valves (112, 212) in each of the first and second filtration paths are used to introduce air into the virus filter (126b, 226b) for pressure hold or air diffusion integrity testing. The flow of the aqueous product-containing fluid is driven by one of two pumps, where the first pump (116a, 216a) is designed for low flow rate operation in a continuous production process and is used to load product from the product inlet (101) onto the filter through the automatic product inlet valve (110a, not shown in the perspective view of FIG. 2) during a continuous process. The second pump (shown as 116b in FIG. 1 but not shown in the perspective view of FIG. 2) is designed for high flow rate operation and is used to perform buffer flushing, equilibration, or sanitization operations for either a batch or continuous process. Here, the second pump (116b) is also used to load product onto the filter during a batch process. A flow meter (shown as 114 in FIG. 1, but not shown in the perspective view of FIG. 2) measures flow rate and provides these data inputs to the controller (140, 240). Pressure transducers (118, 218) at various locations in the system convert pressure to analog electrical signals that are input to the controller (140, 240). Other analytical sensors (120, 220) are placed at or near the product outlet port (203; product outlet port not shown in the perspective view of FIG. 1) of the filter cart system to monitor UV absorbance, conductivity, temperature, and pH, information that is also input to the controller (140, 240).

[0067] In Figure 3, another different embodiment of a portable automated single-use filter cart is illustrated with depth filters (326a) on the outside of the periphery of the cart within two portable accessory modules (with optional filter housings or holders to hold the filters as shown in the embodiment of Figure 3), one depth filter for each of the first and second filtration paths. In this embodiment, portability is optionally enhanced by the presence of both wheels (330a) on the filter cart itself and wheels (330b) on the accessory modules. Each of the two accessory modules shown in Figure 3 has an inlet (334) for the inflow of aqueous product-containing fluid and an outlet (332) for the return of filtered outflow to the periphery of the filter cart.

[0068] In the embodiment shown in FIG. 3, the flow of the aqueous product-containing fluid is driven by one of two pumps, the first pump (316a) is designed for low flow rate operation in a continuous production process and is used to load the product-containing fluid from the product inlet (301) through the automatic product inlet valve (310a) onto the depth filter (326a) and in this embodiment onto the sterilizing grade filter (326c) in the system. In other embodiments, the system can optionally have a virus filtration function installed in the first and second filtration paths, as in the embodiment shown in FIG. 1 and FIG. 2 (126b, 226b). In the embodiment shown in FIG. 3, the second pump (316b) is designed for high flow rate operation and is used to perform buffer flushing, equilibration, or sanitization operations for either a batch or continuous process. Here, the second pump (316b) is also used to load the product-containing fluid onto the filter during a batch process. A flow meter (314) measures the flow rate, and optionally other analytical sensors (320) at or near the product outlet port (outlet port not shown in the perspective of FIG. 3) can provide data input (e.g., for UV absorbance, conductivity, temperature, and pH) to a controller (not shown in the perspective of FIG. 3). Optionally, one or more pressure sensors (318) can be configured at intervals within both the first and second filtration paths.

[0069] For batch production processes, the embodiment shown in Figure 3 includes "crossover" valves (310c, 310d, 310e, 310f) that can automatically switch the flow of product-containing fluid or buffer delivered by the high-flow pump (316b) to a first filtration path (i.e., open 310e and close 310f and 310c) or to a second filtration path (i.e., open 310f and close 310d and 310e) upon instruction from a digital controller. For continuous production processes, the crossover valves can automatically switch the flow of product-containing fluid delivered by the low-flow pump (316a) to a first filtration path (i.e., open 310c and close 310d and 310e) or to a second filtration path (i.e., close 310c and 310f and open 310d). For buffer flushing in a continuous process, the crossover valve can be automatically positioned to direct the flow of buffer from the high flow pump (316b) to the first filtration path (i.e., 310e open, 310f and 310c closed) or to the second filtration path (i.e., 310f open, 310d and 310e closed). In a continuous process, buffer flushing via the high flow pump (316b) can occur simultaneously with the filtration flow of product-containing fluid pumped by the low flow pump (316a). The selection of any one or more buffer inlets (302) and which buffer inlets (302) are operated is automatically controlled by the buffer inlet valve (310b)(es), where the automation of controlling the product inlet valve (310a) and the buffer inlet valve (310b) is under the direction of a controller (not shown in the perspective of FIG. 3). In the embodiment shown in FIG. 3, the pumps (316a, 316b) are operated in a flow control mode using flow meters (314) configured in both the first and second filtration paths.

[0070] Table 1 below provides some useful examples of commercially available mechanical components that can be employed in the construction of the filter cart of the present invention, the examples in Table 1 being merely illustrative and not an exhaustive list of such useful components.

[0071] [Table 1]

[0072] By way of further example, the following numbered embodiments are encompassed by the present invention:

[0073] Embodiment 1: A portable, automated, single-use, replaceable filter cart system for continuous or batch biological product production, comprising: (a) a filter cart for storing, inlet and outlet filters; (a) A portable cart having an outer edge; (b) a valved product inlet within the periphery of the portable cart for receiving an aqueous product-containing fluid from an upstream unit operation outside the periphery of the cart, the upstream unit operation being fluidly connectable to the product inlet by a sealable connector and a connecting line, wherein the product inlet is connected via a connecting line capable of fluidly conveying the aqueous product-containing fluid downstream to a first pump or, interchangeably, a second pump, the first pump being adapted for low flow rate operation and the second pump being adapted for high flow rate operation, and the first pump and the second pump being each configured to alternately pump a flow of the aqueous product-containing fluid into the first filtration path or into the second filtration path; wherein the first filtration path and the second filtration path are respectively as follows: (i) a plurality of filters downstream of the first pump, the plurality of filters being fluidly connected in series along a filtration pathway; and, optionally, one or more portable accessory filtration module(s) outside an outer periphery of the cart, the one or more portable accessory filtration module(s) being fluidly connected to the filtration pathway and capable of receiving the aqueous product-containing fluid therefrom and returning filtered aqueous product-containing fluid thereto; (ii) a plurality of sensors capable of continuously measuring flow rate, flow rate, and / or pressure at one or more locations along the filtration path and transmitting measurement signals to a controller; and (iii) a connecting line capable of fluidly transporting the filtered aqueous product-containing fluid to a downstream product outlet port; Equipped with (c) one or more valved buffer inlets within the periphery of the cart and upstream of the first filtration pathway and the second filtration pathway, each buffer inlet fluidly connected by a connecting line capable of fluidly conveying a desired aqueous fluid received from a reservoir or an upstream unit operation for injection into the first filtration pathway or the second filtration pathway by operation of the first pump or the second pump and the valved buffer inlet in response to a command signal from the controller; and (d) where the controller is configured and programmable to automatically switch the flow of the product-containing fluid from the first filtration pathway to the second filtration pathway in response to a predetermined measurement signal received from the sensor(s) of the first filtration pathway or after a predetermined period of time, where the controller is further configured and programmable to automatically switch the flow of the aqueous product-containing fluid from the second filtration pathway to the first filtration pathway in response to a predetermined measurement signal received from the sensor(s) of the second filtration pathway, where the controller is ... in response to a predetermined measurement signal received from the sensor(s) of the second filtration pathway in response to an indication signal of: (i) operating a first pump during a desired continuous biologics production process, or, interchangeably, operating a second pump during a desired batch biologics production process; (ii) operating the second pump during a desired continuous biological production process to flow a buffer or water through the first filtration path while the flow of the aqueous product-containing fluid passes through the second filtration path, or, interchangeably, to flow a buffer or water through the second filtration path while the flow of the aqueous product-containing fluid passes through the first filtration path; and (iii) controlling a valved product inlet and one or more valved buffer inlets; The device may be further configured and programmed to issue:

[0074] Embodiment 2: The portable, automated, single-use filter cart system of embodiment 1, wherein the controller is configured and programmable for a continuous production process.

[0075] Embodiment 3: The portable, automated, single-use filter cart system of any of embodiments 1-2, wherein the controller is configured and programmable for a batch production process.

[0076] Embodiment 4: The portable automated single-use filter cart system of any of embodiments 1-3, wherein the controller is configured and programmable to interact with a production facility control system.

[0077] Embodiment 5: The portable, automated, single-use filter cart system of any of embodiments 1 to 4, comprising a depth filter.

[0078] Embodiment 6: A portable, automated, single-use filter cart system of any of embodiments 1 to 5, comprising a virus filter.

[0079] Embodiment 7: The portable, automated, single-use filter cart system of any of embodiments 1-6, comprising a sterilizing grade filter.

[0080] Embodiment 8: The portable, automated, single-use filter cart system of any of embodiments 1-7, comprising one or more portable accessory filtration modules.

[0081] The following examples are illustrative and should not be construed as limiting the scope of the invention. EXAMPLES

[0082] Example 1. Depth filter in a continuous process [material and method]

[0083] [Operation Setup] A continuous end-to-end (E2E) process from SUB / perfusion to continuous capture was set up, including a series of two tank viral inactivation filters followed by a series of depth filters. Surge vessels were used between each unit operation to manage flow mismatches between processing steps. Monoclonal antibody (mAb) product was perfused through a 0.2 μm filter as previously described in US Patent No. 5,993,366. Harvesting began on production day 8, and downstream processing was introduced when the surge vessel filled its processing volume.

[0084] 0.55m 2 A 1000-milliliter X0SP depth filter (Millistak+™; Millipore Sigma) was used in conjunction with a SHC 0.2 μm sterilizing grade filter (Millipore Express™; Millipore Sigma) to remove turbidity and process related impurities. Product loading was set for 6 days of operation or until a maximum pressure of 20 psi was reached. Loading can be set by k, volume or time; in this example, time-based switching was used. Production was terminated on day 22, when downstream operations were phased out when the surge vessel filled to minimum volume.

[0085] [Installation] As shown in Figures 1-3, a DF / VF tubing manifold (i.e., tubing system) with probes, sensors, and single-use pump heads with AseptiQuik™ fittings (CPC, Calder Products Company) was installed, and the various parts of the tubing manifold were aseptically connected. Buffer connection lines were aseptically attached to the inlets. The product load inlet was connected to a preceding supply surge vessel. To maintain a closed system, the exiting waste streams were connected together to one waste outlet and connected to a drain by a single-use air break assembly (see U.S. Patent No. 5,331,633). The product outlet port was connected to a collection surge vessel. The depth filter was placed in a POD holder (catalog number MP0DPIL0T, Millipore Sigma), and the lines coming from the filter cart's "Stage 1" (i.e., DF) position to the POD inlet and the lines returning the flow from the POD outlet to the cart were connected using silicone tubing. A 0.2 μm filter was inserted into the "stage 3" (ie, sterilizing grade filter) position and aseptically connected to the tubing manifold.

[0086] [Flushing (cleaning)] The following steps were performed using an automation sequence in the DeltaV Distributed Control System (DCS) automation system (Emerson):

[0087] <Filter Path 1> (i.e., the first filtration path, arbitrarily represented as the upper path in FIG. 4):

[0088] Prime (fill with hydraulic fluid) the inlet connection lines and depth filters to purge air from the system;

[0089] Flush the depth filter with water according to the manufacturer's recommendations, up to the "Stage 1" waste outlet valve (410g in Figure 4), then up to the final waste outlet valve (410j in Figure 4), then to the waste outlet connected thereto;

[0090] Flush with 0.5N NaOH up to the "Stage 1" waste outlet and hold for 30 minutes to disinfect the filter and POD holder connections;

[0091] Flush NaOH with equilibration buffer (EQ) first up to the "stage 1" waste outlet valve (410g), then up to the final waste outlet valve (410j in Figure 4) and the waste outlet connected thereto;

[0092] Allows automation to begin loading the depth filter once the feed surge bin reaches a start weight. The depth filter loading rate is based on the continuous process flow rate to maintain level control in the surge bin.

[0093] The automation allowed the set point of the product loading flow rate to be changed to fast, normal, or slow depending on the tank level in the preceding surge vessel to control the volume in the surge vessel to a predetermined range. An initial buffer was held in the connecting lines to wash the filter and drain it through the waste outlet valve (410j in FIG. 4), then switched to the product recovery outlet valve (410k in FIG. 4), and then to the product outlet port (403). The first filter was loaded for 6 days, and the automation monitored the pressure of the system while continuously controlling the flow rate.

[0094] [Filter Swap] Prior to the end of filter path 1 loading, the depth filter of filter path 2 (i.e., the second filtration path, arbitrarily represented as the lower path in FIG. 4) was prepared in a manner similar to all pre-use flushes performed on the automation-driven filter cart system. The filter path 2 flush was performed simultaneously with the product loading of filter path 1 using the valve configuration shown in FIG. 4. When filter path 1 had 10% of its loading time remaining, the automation activated a flag to initiate a final EQ flush step of filter path 2. When filter path 1 reached the end of its loading time, the automation switched the flow path of the product load from the filter path 1 load to the filter path 2 load and directed it to the waste outlet valve of filter path 2 (410j, lower path in FIG. 4) and to the waste outlet connected thereto. After the switch to product loading on filter path 2 occurred, the automation initiated an EQ recovery wash from the high flow pump (416b) through filter path 1 to the product recovery outlet valve on filter path 1 (410k, upper path in FIG. 4) and from there to the product outlet port (403) to recover product trapped in the filter and plumbing connections. The EQ recovery wash on filter path 1 was performed simultaneously with the product loading on filter path 2. After reaching a predetermined volume transition point, the automation switched filter path 2 from the waste valve (410j, lower path in FIG. 4) to product recovery (410k, lower path in FIG. 4) to complete the operation of filter path 1. The same procedure was performed in the same manner as above with a new set of filters prepared on filter path 1 for filter switch to the third set of depth filters, and switched back to product loading on filter path 1 once the loading of the second depth filter was completed. In this experiment, a total of three depth filters were used in the process, however, alternation between filter path 1 and filter path 2 can continue as many times as desired for a given sequential process.

[0095] [result] [Depth filter (DF) flushing performance] A flow rate of 2.3 liters per minute (L / min) was used for all wash steps. Depth filter prepressure started at 10 psi and was slowly reduced to 8-9 psi during the wash. At the end of the analytical flush, the pH was slightly different between the filter path and the EQ buffer (0.1-0.3 pH units higher than the EQ buffer), which may be due to an offset in the pH probe after autoclaving. It is standard procedure to perform a one-point calibration adjustment on the system pH transmitter to match the probe pH to the offline measurement, but this was not performed in this study. Table 2 summarizes the wash steps and the time and volume required to perform each step. Including transitions, the priming and flushing process took approximately 2 hours.

[0096] [Table 2]

[0097] The first depth filter was loaded with flow over a period of six days. As shown in Figure 5, a pressure increase was observed in each of the filters, and filter replacement occurred sooner than six days in filters 2 and 3 due to increased turbidity of the load. The pressure increase and duration of loading can be optionally modified in future runs by increasing the area of ​​the filter used in the process.

[0098] Example 2. Virus filters in a continuous process [material and method]

[0099] [Operation Setup] The process was from the DF process described in Example 1 (above) with two polishing chromatography steps prior to the virus filter step into the surge vessel. The product was titrated with acid buffer in the preceding loading surge vessel. Harvest harvest began on day 8 of production and the downstream process was phased in when the surge vessel was at operating volume. The process used 1000 L / m 2Planova(TM) 20N virus removal filter (1m 2 Asahi Kasei) was used without replacement. Production was terminated on day 22, and downstream operations were phased out when the surge vessel reached minimum capacity.

[0100] [Installation] A tubing manifold with probes, sensors, and single-use pump heads was installed as described in Example 1 (above), and the various parts of the tubing manifold were aseptically connected together using AseptiQuik™ fittings. The buffer connection line was aseptically connected to the inlet. The product load inlet was connected to a leading feed surge receptacle. The exiting waste streams were connected together to one waste outlet to maintain a closed system, which was connected to a drain by a single-use air break assembly (see US Pat. No. 5,399,133). The product outlet port was connected to a collection surge receptacle. A virus filter was placed in the "Stage 2" (VF) filter holder and aseptically connected to the inlet (feed) and outlet (permeate) tubing manifolds. The permeate was aseptically connected to manually clamped tubing. A 0.2 μm filter was inserted in the "Stage 1" (DF) position as a prefilter and aseptically connected to the tubing manifold.

[0101] [Leak test before use] Although this demonstration run was not leak tested prior to use, the system of the present invention is equipped with a dedicated valve to attach air to pressurize the filter to the required volume and perform a pressure hold test according to the manufacturer's procedures.

[0102] [Flushing (cleaning)] The following steps were performed using an automated sequence in a DeltaV Distributed Control System (DCS) automation system (Emerson).

[0103] <Filter Path 1> (i.e., the first filtration path, arbitrarily represented as the lower path in FIG. 4):

[0104] Prime (fill with hydraulic fluid) the inlet connection lines and depth filters to purge air from the system;

[0105] Rinse the virus filter with water according to the manufacturer's recommendations, which involves purging air through the retentate port; wash through the virus filter, with the permeate flowing to the "stage 2" (VF) waste outlet valve (410g, upper pathway in Figure 4) and from there to the waste outlet;

[0106] Equilibration buffer (EQ) buffer flows through the permeate to the "Stage 2" waste outlet valve (410g, upper path in Figure 4) and then out to the waste outlet connected thereto;

[0107] Once the supply surge vessel reaches the starting weight and the supply tank reaches the pH set point, automation allows loading of the virus filter to begin.

[0108] The pH titration was performed in a feed surge vessel with an acid buffer attached to the titration pump. Automation was used to achieve the pH set point. The automation would pause the viral filter loading if the pH was out of process specification and resume if the pH was back in range. The viral filter loading flow rate was based on the continuous process flow rate to maintain level control in the surge vessel. The automation allowed the product loading flow rate set point to be changed to fast, normal, or slow depending on the tank level in the preceding surge vessel to control the volume in the surge vessel to a predetermined range.

[0109] The first buffer was held in the connecting lines to wash and drain the filter, then the flow was switched to the product outlet port. Automation monitored the pressure of the system, and alarms allowed to alert the operator in case of high pressure or flow rate deviations.

[0110] As described in Example 1, virus filters can be swapped between filter path 1 (i.e., the first filtration path, arbitrarily represented as the upper path in FIG. 4) to filter path 2 (i.e., the second filtration path, arbitrarily represented as the lower path in FIG. 4) and back to filter path 1. Although filter swaps can be performed for longer runs, in this demonstration process only one virus filter was used for the duration of the run and no filter swaps were performed.

[0111] [result] The virus filters were treated continuously for 8 days, during which time the inlet pressure of the Planova™ filter showed a minimal increase from 0.7 to 1.3 psi (Figure 6).

Claims

1. 1. A portable, automated, single-use, replaceable filter cart system for continuous or batch biologics production comprising: (a) a portable cart having an outer perimeter; (b) a valved product inlet within the periphery of the portable cart for receiving an aqueous product-containing fluid from an upstream unit operation outside the periphery of the cart, wherein the upstream unit operation is fluidly connectable to the product inlet by a sealable connector and connecting lines, wherein the product inlet is connected to a first pump, or interchangeably to a second pump, via connecting lines capable of fluidly conveying the aqueous product-containing fluid downstream, wherein the first pump is adapted for low flow rate operation and the second pump is adapted for high flow rate operation, and wherein the first pump and the second pump are each configured to pump a flow of the aqueous product-containing fluid alternately into a first filtration path or into a second filtration path; wherein the first filtration path and the second filtration path each have: (i) a plurality of filters downstream of the first pump, the plurality of filters being fluidly connected in series along the filtration pathway; and, optionally, one or more portable accessory filtration modules outside the periphery of the cart, the one or more portable accessory filtration modules being fluidly connected to the filtration pathway and capable of receiving aqueous product-containing fluid therefrom and returning filtered aqueous product-containing fluid thereto; (ii) a plurality of sensors capable of continuously measuring flow rate, flow rate, and / or pressure at one or more locations along the filtration path and transmitting measurement signals to a controller; and (iii) a connecting line capable of fluidly conveying the filtered aqueous product-containing fluid to a downstream product outlet port; Equipped with (d) one or more valved buffer inlets within the periphery of the cart and upstream of the first filtration pathway and the second filtration pathway, each valved buffer inlet fluidly connected by a connecting line capable of fluidly conveying a desired aqueous fluid received from a reservoir or an upstream unit operation for injection into the first or second filtration pathway by operation of the first pump or the second pump and the valved buffer inlet in response to a command signal from the controller; and (e) wherein the controller is configured and programmable to automatically switch flow of product-containing fluid from the first filtration path to the second filtration path in response to a predetermined measurement signal received from the sensor in the first filtration path or after a predetermined time period, and wherein the controller is further configured and programmable to automatically switch flow of aqueous product-containing fluid from the second filtration path to the first filtration path in response to a predetermined measurement signal received from the sensor in the second filtration path, and wherein the controller is ... in response to an indication signal of: (i) operating the first pump during a desired continuous biologics production process, or, interchangeably, operating the second pump during a desired batch biologics production process; (ii) operating the second pump during the desired continuous biological production process to flush a buffer or water through the first filtration path while a flow of aqueous product-containing fluid passes through the second filtration path, or, interchangeably, to flush a buffer or water through the second filtration path while a flow of aqueous product-containing fluid passes through the first filtration path; and (iii) controlling said valved product inlet and said one or more valved buffer inlets; and Portable automated single-use filter cart system.

2. The control device is configured and programmable for a continuous production process.

10. The portable, automated, single-use filter cart system of claim 1.

3. The control device is configured for a batch production process. The portable automated single-use filter cart system according to any one of claims 1 to 2.

4. The controller is configured and programmable to interact with a production facility control system. The portable automated single-use filter cart system according to any one of claims 1 to 3.

5. Equipped with a depth filter, The portable automated single-use filter cart system according to any one of claims 1 to 4.

6. Equipped with a virus filter, The portable automated single-use filter cart system according to any one of claims 1 to 5.

7. Equipped with sterilizing grade filters, The portable automated single-use filter cart system according to any one of claims 1 to 6.

8. comprising said one or more portable accessory filtration modules; The portable automated single-use filter cart system according to any one of claims 1 to 7.

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