Depth filter and virus filter (DF / VF) cart 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 proteins.

Background Art

[0002] The biopharmaceutical industry is undergoing significant changes, partly driven by the rapid increase in approvals of new biotherapeutics, the improvement of protein expression rates, and the increasing pressure from the biosimilars market (Non-Patent Document 1).

[0003] The rapid increase in the pharmaceutical market share of biological agents (from 11% in 2002 to approximately 20% in 2017) and the need for access to affordable pharmaceuticals in developing regions worldwide have led to a demand for the development of rapid, sustainable, and cost-effective production methods (Non-Patent Document 2).

[0004] According to the production technology of biological agents that replaces conventional batch processing platforms, it is possible to utilize major advantages such as improved processing capacity, operational flexibility, cost reduction, and reduction of environmental impact due to reduction of installation area. A bioprocessing plant is designed to include a continuous production process in which upstream and downstream unit operations are integrated, enabling rapid equipment conversion, flexibility in products and volumes, and reduction of production costs compared to batch culture processing (see, for example, Non-Patent Documents 3, 4, 5, and 6).

[0005] The emergence of continuous perfusion technology has supported greater progress in connecting upstream process equipment for operation in continuous mode. This processing strategy has been valuable for some companies over the past 25 years and has helped to overcome stability problems related to products (Non-Patent Document 7).

[0006] Recent cell lines and media are designed to achieve higher cell densities, particularly in contrast to fed-batch processing, with some cultures achieving viable cell densities exceeding 100 million cells / mL (Non-Patent Literature 8). As a result, the bottleneck in typical biopharmaceutical production facilities has shifted from the production bioreactor (upstream process) to the purification train (downstream process), and especially to the chromatography column due to its dimensional limitations. The increasing amount of protein derived from current production cell lines makes purifying the large batch-sized products produced a challenging task (Non-Patent Literature 9). Thus, the issue of integrating upstream and downstream biopharmaceutical production processes continues to plague the biopharmaceutical production industry.

[0007] Warikoo et al. have reported achieving reductions in column size and buffer usage by integrating a continuous capture chromatography process downstream of the production bioreactor (Non-Patent Document 10).

[0008] Godawat et al. have demonstrated that end-to-end continuous bioprocesses are feasible, but still face several challenges, including the development of robust virus clearance and automation strategies to ensure high product quality (Non-Patent Document 11).

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

[0010] Equipment and systems conventionally known in the art, including filter carts, have been insufficient for modern biological drug production lines employing flexible configurations (see, for example, Patent Documents 2, 3, and 4).

[0011] Therefore, there is a need for a dual-pass depth filter and viral filter (DF / VF) cart that can efficiently and flexibly meet the production process requirements for batch and / or continuous biological products (biopharmaceuticals) with compatibility over long production periods. This invention provides such a solution. [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 bioreactor™-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 System, Equipment, and Process, International Publication No. 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 Using the Same, U.S. Patent Application Publication No. 2020 / 384380 [Patent Document 4] Reinbigler et al., Equipment for Treating Biological Fluids, U.S. Patent Application Publication No. 2012 / 0248025 [Summary of the Invention]

[0014] The present invention includes a portable automatic single-use filter cart system for the production of continuous or batch-exchangeable biologies.

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

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

[0017] (b) A valve-equipped biota inlet within the outer edge of the cart for receiving an aqueous biota-containing fluid derived from an upstream unit operation outside the outer edge of the portable cart, where the upstream unit operation is fluidly connectable to the biota inlet by a sealable connector and a connecting line, and the biota inlet is connected to a first pump or alternatively to a second pump via a connecting line capable of fluidly transporting the aqueous biota-containing fluid downstream, the first pump being adapted for low-flow operation and the second pump being adapted for high-flow operation, the first pump and the second pump being each configured to push the flow of the aqueous biota-containing fluid into a first filtration path or alternatively into a second filtration path, and the first filtration path and the second filtration path each comprise the following (i)-(iii):

[0018] (i) A plurality of filters downstream of the first pump, the plurality of filters being fluidly connected in series along the filtration path; and optionally, one or more portable accessory filtration modules outside the outer edge of the cart, fluidly connected to the filtration path, capable of receiving the aqueous biota-containing fluid therefrom and returning the filtered aqueous biota-containing fluid thereto; (ii) A plurality of sensors capable of continuously measuring the flow rate, flow volume, and / or pressure at one or more positions along the filtration path and transmitting a measurement signal to a control device; and (iii) A connecting line capable of fluidly transferring the filtered aqueous biota-containing fluid to a downstream biota outlet port.

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

[0020] (c) One or more valved buffer inlets located within the outer edge of the cart and upstream of the first and second filtration paths, each buffer inlet being fluidically connected by a connecting line capable of fluidly transporting a desired aqueous fluid received from a reservoir or upstream unit operation for injection into the first or second filtration path by the operation of the first or second pump and the valved buffer inlet in response to an instruction signal from the control device;

[0021] (d) The control device is configured and programmable to automatically switch the flow of the 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 the aquatic 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) Activate the first pump during a desired continuous biological product production process, or, interchangeably, activate the second pump during a desired batch biological product production process; (ii) During a desired continuous biological product production process, a second pump is operated to allow buffer or water to flow through the first filtration path while the aquatic product-containing fluid flows through the second filtration path, or interchangeably, to allow buffer or water to flow through the second filtration path while the aquatic product-containing fluid flows through the first filtration path; and (iii) Control the 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 automatic single-use DF / VF filter cart of the present invention assists in switching between a first filtration path and a second filtration path, without limit, as required by the execution of the process, even if some manual intervention is required to replace the filter as needed.

[0023] The above 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. The entire Spectrum is intended to be considered as a unified disclosure, and it should be understood that all combinations of features described herein are contemplated, even if combinations of features are not found together in the same sentence, paragraph, or section of the Spectrum. For example, certain aspects of the Invention described as a genus, and all members of the genus, should be understood to be aspects of the Invention individually. [Brief explanation of the drawing]

[0024] [Figure 1] Figure 1 shows an embodiment of a portable automatic single-use filter cart system. [Figure 2] Figure 2 shows the embodiment of Figure 1 from a different viewpoint. [Figure 3] Figure 3 shows another embodiment including an “offskid” filtration module. This embodiment of the portable automatic single-use filter cart is shown with depth filters (326a) housed in housings or holders on the outer edge of the cart within two portable accessory filtration modules, one for each of the first and second filtration paths. [Figure 4] Figure 4 shows a schematic diagram of the design of the portable automatic single-use DFVF filter cart system of the present invention, which can operate either a depth filter (426a) or a virus filter (426b) in either batch mode or continuous mode. [Figure 5]Figure 5 shows the changes over time in the load inlet pressure and load turbidity of the depth filter during a typical operation using a depth filter, as described in Example 1 of this specification. [Figure 6] Figure 6 shows the time-dependent virus filter load flow rate and differential pressure in a typical operation using a virus filter, as described in Example 2 of this specification. [Modes for carrying out the invention]

[0025] The headings used herein are for structural purposes only and should not be construed as limiting the subjects described.

[0026] [Definition] Unless otherwise defined herein, scientific and technical terms used in connection with this application shall have meanings generally understood by those skilled in the art. Furthermore, unless otherwise specified in the context, singular terms shall include plural forms, and plural terms shall include singular forms. Thus, where used herein and in the appended claims, the singular forms "a," "an," and "the" shall include multiple references unless it is clearly indicated in the context that they are inconsistent. For example, a reference to "a protein" includes multiple proteins, and a reference to "a cell" includes a collection of multiple cells.

[0027] The present invention encompasses a portable, automated, single-use filter cart system for interchangeable, continuous, or batch-type biological product production. The filter cart of the present invention can be operated under sterile conditions and can be incorporated, as needed, into an automated, controlled, and regulated biological product production process, regardless of whether the production process (hereinafter, the terms process, treatment, and procedure are interchangeable) is a batch process, a semi-continuous process, or a continuous process. The filter cart system of the present invention can function interchangeably in any of these treatment modes and can be positioned between different unit operations as needed in a particular production process.

[0028] The term “integrated” in relation to a process for producing purified proteins or other biological products of interest (but not limited to, for example, protein pharmaceuticals) means that one or more upstream and / or downstream processes in the production process are performed under common or coordinated control based on programmed commands modified by current sensor feedback of parameters defined with respect to setpoints 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 the efficiency or harmony of their functions toward a single purpose.

[0029] In a typical biological product production process, after collecting a cell culture-containing fluid with the protein of interest or other biological product (e.g., an antibody), the product can be further purified from the cell-free supernatant fraction. Typically, protein purification is usually achieved by a series of arbitrary chromatographic steps such as anion exchange chromatography, cation exchange chromatography, affinity chromatography (using protein A, protein G, or protein L as affinity ligands), hydrophobic interaction chromatography, hydroxyapatite chromatography, and size exclusion chromatography. Furthermore, the purification process may include one or more deep filtration, viral filtration, ultrafiltration, nanofiltration, or dialysis filtration steps, and / or, optionally, an acidic viral inactivation step. Depending on the biological product being recovered, any other known techniques for protein purification such as ethanol precipitation, reverse-phase HPLC, chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are also possible.

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

[0031] The “continuous” form or mode of a production process or system means a processing mode in which a perfusion bioreactor is fluidically connected (directly or indirectly via intervening unit operations) to a continuous capture chromatography process (e.g., processing by the first chromatography system) in an uninterrupted flow from the bioreactor to a first chromatography system, followed by fluidically connected in an uninterrupted flow to a downstream virus inactivation step, and optionally fluidically connected in an uninterrupted flow to a depth filter. Further downstream product purification steps (e.g., processing by a second chromatography system, an optionally third chromatography system, a virus filter, and ultrafiltration / dialysis filtration) are fluidically connected to the above upstream processing steps in a continuous, uninterrupted flow, with the intervening of optionally surge vessels.

[0032] The “semi-continuous” form or mode of the production process refers to a processing mode in which a perfusion bioreactor is fluidly connected in an uninterrupted flow to a continuous capture chromatography process (e.g., processing by a first chromatography system), and to processing by another step, such as a virus inactivation system, but not limited to this, and optionally to a depth filter in an uninterrupted flow, storing the virus-inactivated product pool in a retention container before further processing. The temporary storage of the product pool in the retention container is then followed by one or more batch downstream processing steps, which may be fluidly connected in an uninterrupted flow, for example, to processing by a second chromatography system, an optionally third chromatography system, and ultrafiltration / dialysis filtration, with the interposition of a surge container or retention container (i.e., retention container if there are two or more batch processes or operations).

[0033] A "perfusion bioreactor" is a cell culture bioreactor that allows for the addition and removal of an equivalent volume of culture medium from the reactor while retaining cells within the bioreactor. A perfusion bioreactor comprises a bioreactor and an operablely attached perfusion system. This perfusion system provides a stable source of fresh nutrient medium and removal of cellular waste. Typically, the bioreactor and perfusion system of a perfusion bioreactor may be separate mechanical units that work in coordination. However, there are many commercially available examples, though not limited to them, such as the Xcellerex® brand single-use bioreactors (SUBs from GE Healthcare Life Sciences) and the KrosFlo® brand perfusion flow path assemblies and systems (Spectrum from Repligen), and these bioreactors and perfusion systems can be appropriately combined into a single perfusion bioreactor by those skilled in the art. Alternatively, the bioreactor and perfusion system can be assembled into a single mechanical unit, for example, but are not limited to this, such as the 3D Biotek brand perfusion bioreactor (manufactured by Sigma-Aldrich). Protein products secreted in the bioreactor can be continuously collected by microfiltration during the process of removing the culture medium via the perfusion system, and the target protein is isolated in the microfiltered filtrate thus exiting the perfusion system.

[0034] A process in a production line or a system within an automated production facility is "fluidically" executed if the material containing the target protein flows between processes or systems via pipes, tubes, or other closed conduits without manual loading or unloading, or if it is "fluidically connected" to another process or system in a production line, or if material is "fluidically" acquired from another process or system in a production line. Such pipes, tubes, or other closed conduits between processes or systems are considered "connection lines" if they can fluidly transport an aquatic product-containing fluid downstream within a system or between systems or unit operations. Connection lines can be made from any suitable non-reactive, non-porous material, such as, but not limited to, silicone rubber, Teflon®, or stainless steel. Aseptic connectors are useful for joining connection lines to other components, as is known to those skilled in the art.

[0035] A single step in a production process or a system within an automated production facility is generally called a “unit operation.” A “production line” or “production processing line” (as used interchangeably herein) is a series of unit operations involved in the production of a biological product, such as a protein of interest, a pharmaceutical raw material, or a formulation, but is not limited to these terms.

[0036] A “batch unit” or “skid” is a unit operation. Typical examples include single-use bioreactors, perfusion systems, first chromatography systems, any second chromatography systems, any third chromatography systems, and / or ultrafiltration / dialysis filtration systems, each of which may, but may not, be configured as a “skid.” Virus inactivation systems and, if necessary, neutralization systems can also be “skids.” Batch units are configured to communicate with a Process Control System (PCS) or Supervisory Control and Data Acquisition (SCADA) system (e.g., by hardwiring, fiber optic cables, or wireless connections). The portable automated single-use filter cart system of the present invention can also be configured as a “skid” within a production process. A “non-batch unit” is a device that supports one or more batch units. Non-batch units are configured to communicate with a PCS or SCADA system (e.g., by hardwiring, fiber optic cables, or wireless connections). Furthermore, dongles and / or Profibus devices, or similar digital information storage devices and electronic hardware connectors(s), can be attached to non-batch units to identify their location (location relative to batch units) and role (e.g., supply tank, collection tank, filter bank, etc.). For convenience and flexibility, filter banks, heat exchangers, surge vessels, supply tanks, reservoirs, holding vessels, recovery vessels or collection tanks (e.g., elution recovery vessels), and optionally portable mixers and other mixing vessels are typically configured as non-batch units, although in some embodiments, such unit operations may also be included in a “skid” including control by hardwiring, fiber optic cables, or wireless connectivity.

[0037] The terms “automated,” “automatically controlled,” or “automatically” are used interchangeably in relation to a production process or production equipment and mean the implementation or execution of one or more processing steps, or computer control over the operation of components or systems of production equipment, which optionally involves feedback control of the process or operation. Typically, a computer control unit receives digital signals from detectors of physical or chemical parameters to be controlled and sends digital instructions of response to the system or subsystem.

[0038] The terms “single-use” or “single-use” as used interchangeably herein mean that a component of a particular sterile production line, i.e., a part of a sterile apparatus, such as a component used in the portable automatic single-use filter cart system of the present invention, is constructed or configured to be used for a single production run (however, it may be reused if quality and sterile hygiene can be guaranteed for multiple production runs). The single-use component is then discarded and can be replaced for subsequent production runs with another single-use component consisting of the same or modified configuration without requiring cleaning (flushing) and disinfection of the component between production runs. Such single-use components are commercially constructed and available. 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, sterile-grade filters, or connectors, connecting lines or valves, containers or bags. Single-use depth filters, single-use virus filters, single-use pre-filters, and single-use sterile-grade filters are widely available commercially and include, for example, single-use filter assembly systems including filters (various membranes and pore sizes 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 filter (SU-VFS; Asahi Kasei Bioprocess America, Inc.), or Viresolve® Pro virus filter (MilliporeSigma).Single-use connection 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, and the connection line system is also known as a "tube manifold." Single-use sterile connectors are available from various manufacturers, including AseptiQuik® connectors (Colder Products Company), Kleenpak® Presto sterile connectors (Pall Biotech); and Lynx® ST connectors (MilliporeSigma).

[0039] The terms “switching” or “switch,” as used interchangeably herein with respect to the first and second filtration paths, mean changing the direction of flow of an aquatic product-containing fluid from the first filtration path to the second filtration path, or vice versa. Such switching can be performed under the automatic control and adjustment of a computer, i.e., control devices (140, 240, 440) and mechanisms (plural), such as valves (110, 210, 310, 410) and pumps (116a, 116b, 216a, 316a, 316b, 416a, 416b), and optionally, under the control of an external control system, such as an automated production equipment control system. While switching can be controlled and adjusted manually, "automatic switching" means that the flow direction switching does not require manual input from the operator, but is instead controlled by a control device (140, 240, 440), which performs the flow direction switching under predetermined criteria or setpoints.

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

[0041] The terms “sterile” or “aseptic,” as used herein, refer to materials that are free from or essentially free from microbial and / or viral contamination. In this regard, “contaminant” means any substance different from the desired component in a preparation, such as cell culture media, components of cell culture media, or aquatic product-containing fluids, 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 more specifically, "batch filtration," or filtration performed in a batch manner, is a process in which a specific total amount or volume of a preparation, such as cell culture medium or at least one component of cell culture medium, or an aquatic product-containing fluid, or any other preparation, is filtered in a batch, depending on the filter capacity, through a filter such as a depth filter or a virus filter, and the filtration process is finalized before the filtrate is directed to or supplied to a process in which it is used or consumed.

[0043] The terms “continuous filtration,” “online filtration,” or “inline filtration” refer to a filtration process in which a specific total amount or volume of a preparation, which is a cell culture medium or at least one component of a cell culture medium, or an aquatic product-containing fluid, or any other preparation, is continuously filtered through a filter, such as a depth filter or a virus filter, in a manner dependent on the capacity of the filter, and the filtration process continues even when the filtrate is directed towards or supplied to a process in which 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 can be added to the culture after cells have been inoculated into the cell culture medium and cell proliferation has begun. "Cell culture" refers to the cell culture supernatant and the product-producing cells cultured therein. Mammalian product-producing cells are typically cultured at 37±1°C.

[0045] To “cultivate” or “maintain” a given temperature or other conditional parameters (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 may include such automatic adjustments to maintain desired conditions. Cultivation or unit operation conditions such as temperature (not necessarily, but typically about 37°C), pH, and oxygenation are maintained at the desired setpoint by a digital control unit (DCU) and sensory monitor, which are commercially available or can be constructed by those skilled in the art. Digital control units (DCUs) capable of controlling and monitoring the state of unit operations within a production process are manufactured and commercially available by companies such as BBraunn, New Brunswick, or Sartorius.

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

[0047] The term "buffer" or "buffer solution" refers to a solution that resists changes in pH due to the action of its conjugate acid-base range. Examples of useful buffers for controlling pH in the range of approximately pH 4 to pH 8 include phosphates, bicarbonates, acetates, MES, citrates, Tris, bis-Tris, histidine, arginine, succinates, citrates, glutamates, and lactates, or combinations of two or more of these, or other mineral or organic acid buffers. Salts containing sodium cations, ammonium cations, and potassium cations are often used when preparing buffer solutions.

[0048] The term “loading buffer” or “equilibrium buffer” refers to a buffer and salt(s) mixed with a product-containing preparation (e.g., supernatant or filtrate of a batch or perfusion culture, or an elution pool containing the biological product of interest, e.g., an antibody) for loading a protein preparation into an affinity purification matrix, or, in some cases, into a depth filter or viral filter. Examples of affinity purification matrices include protein A or protein g conjugate matrices, or specific target ligand conjugate affinity chromatography matrices. This buffer is also used to equilibrate applicable matrices or filters before loading and for washing after loading the biological product.

[0049] [cart] The portable automatic single-use filter cart system of the present invention has a perimeter or boundary and includes a frame, top, base, and preferably wheels, spheres, sleds, casters, or other suitable components at the base to enhance the portability of the cart (see, for example, (130) in Figure 1, (230) in Figure 2, and (330a) in Figure 3). Wheels, spheres, sleds, casters, or other suitable components to enable or enhance portability may also be included at the base of the accompanying filtration module (see, for example, (330b) in Figure 3). The cart frame, top, base, and any optional shelves, trays, drawers, hooks, clips, snaps, and piping included in the cart are considered to be located together with the perimeter of the cart. In different embodiments, the shape of the cart can be modified to be convenient for a desired application or purpose in a production line, but generally, a rectangular or cubic three-dimensional shape is often the most convenient for many applications (see, for example, Figures 1, 2, and 3). The cart can be constructed from any suitable material, but is not limited to, stainless steel, aluminum, titanium, metal alloys, plastics, or any other material with suitable strength, durability, washable and / or disinfectable properties, or a combination of these materials. In one embodiment, 316L stainless steel is used to construct the cart due to its corrosion resistance and washability. The size of the cart can be modified as needed for a particular purpose. In some typical embodiments, the dimensions of the cart's outer rim are approximately 65 inches x 32 inches x 45 inches (length x width x height).

[0050] In some embodiments, the control device is located within the outer edge of the cart, for example, within the frame boundary (see, for example, 140 in Figure 1 and 240 in Figure 2), or on top of the cart. In other embodiments, the control device may be located outside the outer edge of the cart itself, insofar as it can communicate with sensors on the pump, valve, and filter cart; that is, insofar as the control device can receive measurement signals transmitted from the sensors and transmit instructions to all pumps and valves. Such transmission and / or reception of measurement or instruction signals may, in some cases, be done via electrical cables, wiring, fiber optic cables, or via infrared or radio wave transmission and / or any other suitable medium.

[0051] In some embodiments, hooks (e.g., (170) in Figure 1 and (270) in Figure 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 transmission cables, or wiring, and / or for management or support of tubing (flexible piping) or plumbing (rigid piping). For example, trays or hooks for holding tubing or cables or wiring that protrude from the outer edge of the system can improve and maintain tidiness. In some embodiments, several small plastic clips are attached to the front and back of the cart (or other vertical cart section) to hold tubing in place.

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

[0053] [valve] The automatic valves of the portable automatic single-use filter cart system of the present invention can be any suitable type for regulating the flow of fluid through the connection line. In some embodiments, the automatic valves are robot-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 tube to pinch and close the tube of the connection line, are useful embodiments (e.g., pinch valves manufactured by Aquasyn LLC or Acro Associates (a wholly owned subsidiary of Bimba Manufacturing Corporation)). Various pinch valves are useful, but are not limited to overmolded tube assemblies for reducing dead volume in the tube. Other useful options include valve redundancy, such as 3-way or 4-way structures. Those skilled in the art will know how to select the valve best suited to a given embodiment, taking into account the desired level of simplification, cost-effectiveness, and the nature of the selected tube assembly, in terms of minimizing the valve footprint and dead volume.

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

[0055] However, in some embodiments, the control devices (140, 240, 440) may also be configured and programmed to interact with a comprehensive production equipment control system. Generally, the production equipment control system can be used to control an automated production line having a flexible configuration that includes individual unit operations, such as the portable automated single-use filter cart system of the present invention. That is, the production equipment control system can be designed to adapt to 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 containing equipment from the original manufacturer, such as a single-use bioreactor system, a perfusion system, or a continuous chromatography system. The skid may also include a flow control device, such as a pump. 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 can be achieved using electrical cables. The electrical cables may be configured to accommodate serial communication, bus communication, or Ethernet® communication. In some embodiments, the electrical wiring is of the recommended standard 232 (RS-232) wiring. It can be done this way.

[0056] The portable portion of the equipment, such as the portable automatic single-use filter cart system of the present invention, may include, or may be otherwise coupled to, a network gateway hardware device that enables communication between the skid and the production equipment control system. Furthermore, at least some of the skids can be logically configured to be coupled to various portable devices. In this way, a portion of the portable equipment can be physically connected to a specific 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 portions of equipment coupled to the skid.

[0057] The location of the portable automatic single-use filter cart system of the present invention for depth filter or virus filter operation on a production line can be assigned and / or identified by the production equipment control system. Furthermore, specific sequence logic for specific depth filter or virus filter processing is also smoothly executed by the production equipment control system in the form of automated recipes or methods compliant with ISA-88 and ISA-95 (and / or other applicable) standards to control one or more control modules, flags, and / or status identifiers for depth filters and / or virus filters, and to execute one or more control modules simultaneously with the filter cart. The production equipment control system can monitor various processing conditions and processing signals, such as pressure or flow rate.

[0058] For example, the pressure within the filter assembly of the filter cart can be monitored based on pressure values ​​obtained from pressure sensors (118, 218, 318, 418) included in the filter cart assembly and relayed by filter cart control devices (140, 240, 440). The production equipment control system can determine when the pressure within the first filter assembly through which the material flows has reached at least a threshold level. The pressure threshold level can indicate that the filters included in the first assembly need to be replaced due to a decrease in the amount of material that can be processed by the filters, and can issue instruction signals and / or alarm signals. The production equipment control system can then send signals to the control devices (140, 240, 440) to switch the flow of aquatic product-containing fluid from the first filtration path to the second filtration path by operating a valve, as illustrated in more detail in the schematic diagram of Figure 4. If necessary, for example, over long-term production operation, the filters included in the first and / or second filtration paths can be replaced. Similarly, the control unit can be programmed to respond to other preset parameters (e.g., duration, volume throughput) or trigger conditions measured by analytical sensors (120, 220, 320, 420) that prompt the control unit to switch the flow of aquatic bio-containing fluid from one channel to the other, and / or to respond to operator prompts (i.e., manual intervention).

[0059] [System design and exemplary embodiments] Figure 4 shows a schematic diagram of the design of the portable automated single-use DFVF filter cart system of the present invention, which allows either the depth filter (426a) in so-called "stage 1" mode or position, or the virus filter (426b) in so-called "stage 2" mode or position, to operate in either batch mode or continuous mode. As further shown in Figure 4, the system design can accommodate various configurations of pre-filters, sterile-grade filters (426c), and / or depth filters (426a), and / or virus filters (426b), which are placed on the filter cart (see, for example, 126b in Figure 1), or optionally placed off the outer edge of the filter cart (indicated as "P" in Figure 4). These filters can be installed at three different positions, also called "stages," on the first or second filtration path. "Stage 1" refers to a first position (426a) in the filter path, "Stage 2" refers to a second position (426b) in the filter path, and "Stage 3" refers to a third position in the filter path. For example, in some embodiments, the depth filtering process can be such that the depth filter (426a) is placed at Stage 1, located outside the outer edge ("P") of the filter cart system (see also Figure 3), and optionally, a sterile-grade filter (426c) can be placed at Stage 3 on the filter cart schematically shown in 426c (i.e., inside "P"; see, for example, 126c in Figure 1). In this example, the position of Stage 2 (426b) is bypassed by using a piece of tubing to connect the rest of the tubing manifold section instead of a filter to complete the flow path. In an exemplary virus filtering process, a pre-filter may be placed on stage 1 on the filter cart (i.e., within the outer edge "P") as shown by 426a, a virus filter (426b) may be placed on stage 2 either on the filter cart (i.e., within "P") or outside the outer edge (426b), and a sterile-grade filter may be placed on stage 3 on the filter cart system (426c; within "P").In the case of a continuous production process (process) that involves filter replacement, the filter can be placed in either the first filtration path, the second filtration path, or both.

[0060] The system also includes a set of inlets (401, 402) comprising (i) one or more valved buffer inlets (402) and (ii) a product inlet (401) for receiving aquatic product-containing fluid from an upstream unit operation fluidly connected to it 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] A first pump (416a) is designed for low-flow operation in a continuous production process and is used to load product-containing fluid onto the filter from a product inlet (401) through a product inlet valve (410a) during continuous processing. A second pump (416b) is designed for high-flow operation and is used to perform buffer flushing, equilibration, or sanitizing operations for either batch or continuous processing, and the second pump (416b) is also used to load products onto the filters (426a, 426b, 426c) during batch processing. The system also includes a pair of inlets (401, 402) including (i) one or more valved buffer inlets (402) and (ii) a product inlet (401) for receiving aquatic product-containing fluid from an upstream unit operation fluid-connected to it upstream of the first and second filtration paths. The product-containing fluid is fluidically delivered via tubing to the product inlet (401), or the buffer(s) used are fluidically delivered via tubing to one or more buffer inlets (402). 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 one or more 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 buffer inlet valve (410b) is under the direction of a control unit (440). In the schematic diagram of Figure 4, the control device (440) is shown to be outside the outer edge ("P") of the cart, but this is merely for convenience and to clarify the drawing, and the control device may be located inside the outer edge of the cart (see, for example, Figures 1(140) and 2(240)), or, in some cases, outside the outer edge of the cart, insofar as measurement signals and / or indicator signals can be easily exchanged between the analysis sensors (418, 420), valves (410), pumps (416a, 416b), and the control device (440).The pumps (416a, 416b) can operate in either (i) a flow control mode using a flow meter (414) configured in both the first and second filtration paths, or (ii) a pressure control mode using one or more spaced pressure sensors (418) configured in both the first and second filtration paths. For example, in Figure 4, the pressure sensors (418) are positioned in both the first and second filtration paths before the first filter position (i.e., 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 sterile 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 predetermined and configured setpoint. 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 difference across the virus filter (426b) is used to achieve a predetermined and configured setpoint in the automation, using pressure sensors positioned before and after the second filter location.

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

[0064] During buffer flushing, the buffer can be flushed (flowed in large quantities) for the Stage 1 or Stage 2 filter using waste outlet valves (410g) configured in both the first and second filtration pathways. While filtration of the product is initiated, the residual volume in the first and second filtration pathways can be flushed and discarded by flowing through valves (410i), (410g), and / or (410j) to the waste ports (the waste ports are not shown in Figure 4, but see, for example, (104) in Figure 1 and (204) in Figure 2). If the product is switched to product collection through product outlet ports (103, 203, 403) toward downstream unit operations, one of the product outlet valves (410k) may be opened to product outlet port (403). A manual valve (412) is used to introduce air into the virus filter (426b) for pressure holding or air diffusion integrity testing.

[0065] The analytical sensors (420a, 420b, 420c, 420d) are positioned 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 start or stop buffer flushing or product recovery.

[0066] Figures 1 and 2 show embodiments of a portable automatic single-use filter cart system from different viewpoints. Wheels (130, 230) and handles (150, 250) for maneuvering the filter cart's position allow for optional enhancement of portability. One or more additional optional buffer inlets (shown as 102 in Figure 1, but not shown in the perspective view of Figure 2) are present on the filter cart for introducing buffer(s) or water into a first or second filtration path to perform buffer flushing or biofiltration 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 virus filter (e.g., 126b, 226b), and / or a sterile-grade filter (126c, 226c). In the viewpoints of Figures 1 and 2, the first filtration path is shown as an upper path further from the viewer, and the second filtration path is shown as a lower path closer to the viewer, both located at the top of the cart (within the outer edge of the cart). The system employs automatic valves (110, 210) controlled by control devices (140, 240) to guide the flow of aquatic product-containing fluid transported through the connecting lines of the tubes. Specifically, the initial flow enters the system from the upstream unit operation via product inlets (101, 201) and through an automated product inlet valve (110a), while optionally, one or more buffer inlets (102; not shown in the perspective view of Figure 2) transport buffer or water into the system via an automated buffer inlet valve (110b; not shown in the perspective view of Figure 2). In this embodiment and many other embodiments of the present invention, the automatic valves (110, 210) are pneumatic valves and are robotically operated by compressed air transported by a separate air piping system within the cart. Any suitable compressed air source can be employed. Figures 1 and 2 show air intakes (161, 261) for the compressed air piping system, and the perspective view in Figure 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 filters (126b, 226b) for pressure holding or air diffusion integrity testing. The flow of the aquatic product-containing fluid is driven by one of two pumps, where the first pump (116a, 216a) is designed for low-flow operation in a continuous production process and is used to load the product onto the filter from the product inlet (101) through the automatic product inlet valve (110a, not shown in the perspective view of Figure 2) during the continuous process. The second pump (shown as 116b in Figure 1, but not shown in the perspective view of Figure 2) is designed for high-flow operation and is used to perform buffer flushing, equilibration, or sanitizing operations for either a batch or continuous process. Here, the second pump (116b) is also used to load the product onto the filter during a batch process. A flow meter (shown as 114 in Figure 1, but not shown in the perspective view of Figure 2) measures the flow rate and provides this data to the control unit (140, 240). Pressure transducers (118, 218) located at various points in the system convert pressure into analog electrical signals and input them to the control unit (140, 240). Other analytical sensors (120, 220) are located at or near the product outlet port (203; the product outlet port is not shown in the perspective view of Figure 1) of the filter cart system to monitor UV absorbance, conductivity, temperature, and pH, and this information is also input to the control unit (140, 240).

[0067] Figure 3 illustrates another different embodiment of the portable automatic single-use filter cart, comprising two portable accessory modules (having optional filter housings or holders for holding filters, as shown in the embodiment of Figure 3) with depth filters (326a) located outside the outer edge of the cart, 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 aquatic organism-containing fluid and an outlet (332) for returning the filtered outflow to the outer edge of the filter cart.

[0068] In the embodiment shown in Figure 3, the flow of the aquatic product-containing fluid is energized by one of two pumps, the first pump (316a) being designed for low-flow operation in a continuous production process, and is used to load the product-containing fluid into the system from the product inlet (301) through the automatic product inlet valve (310a) onto the depth filter (326a), and in this embodiment onto the sterile-grade filter (326c). In other embodiments, there may optionally be a virus filtration function installed in the first and second filtration paths, as in the embodiments shown in Figures 1 and 2 (126b, 226b). In the embodiment shown in Figure 3, the second pump (316b) is designed for high-flow operation and is used to perform buffer flushing, equilibration, or sanitizing 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) located at or near the product outlet port (the outlet port is not shown in the view of Figure 3) can provide input data (e.g., UV absorbance, conductivity, temperature, and pH) to a control device (not shown in the view of Figure 3). Optionally, one or more pressure sensors (318) can be configured at intervals within both the first and second filtration paths.

[0069] In a batch production process, the embodiment shown in Figure 3 includes “crossover” valves (310c, 310d, 310e, 310f) that, upon instruction from a digital control device, can automatically switch the flow of product-containing fluid or buffer transported by a high-flow pump (316b) to either a first filtration path (i.e., opening 310e and closing 310f and 310c) or a second filtration path (i.e., opening 310f and closing 310d and 310e). In a continuous production process, the crossover valves can automatically switch the flow of product-containing fluid transported by a low-flow pump (316a) to either a first filtration path (i.e., opening 310c and closing 310d and 310e) or a second filtration path (i.e., closing 310c and 310f and opening 310d). In buffer flushing in a continuous process, the crossover valve can be automatically positioned to direct the buffer flow from the high-flow pump (316b) to either a first filtration path (i.e., opening 310e and closing 310f and 310c) or a second filtration path (i.e., opening 310f and closing 310d and 310e). In a continuous process, buffer flushing via the high-flow pump (316b) can be performed simultaneously with the filtration flow of the 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)(or more), where the automation of controlling the product inlet valve (310a) and buffer inlet valve (310b) is performed under the direction of a control device (not shown in the viewpoint of Figure 3). In the embodiment shown in Figure 3, the pumps (316a, 316b) are operated in flow control mode using a flow meter (314) configured in both the first and second filtration paths.

[0070] Table 1 below shows some useful examples of commercially available mechanical components that can be used in constructing the filter cart of the present invention. The examples in Table 1 are merely illustrative and not an exhaustive list of such useful components.

[0071] [Table 1]

[0072] Further examples include the following numbered embodiments, which are included in the present invention.

[0073] Embodiment 1: A portable automated single-use filter cart system for replaceable continuous or batch production of biological products, comprising (a) to (d) below: (a) Portable cart having an outer edge; (b) A valved product inlet within the outer edge of a portable cart for receiving aquatic product-containing fluid from an upstream unit operation outside the outer edge of the cart, the upstream unit operation being fluidically connected to the product inlet by a sealable connector and connecting line, the product inlet being connected via a connecting line capable of fluidizing the aquatic product-containing fluid downstream toward a first pump or interchangeably toward 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 and second pumps being configured to alternately pump the flow of aquatic product-containing fluid toward a first filtration path or a second filtration path, Here, the first filtration pathway and the second filtration pathway are as follows: (i) a plurality of filters located downstream of the first pump, the plurality of filters being fluidically connected in sequence along the filtration path; and optionally, one or more portable accessory filtration modules located outside the outer edge of the cart, fluidically connected to the filtration path, capable of receiving aquatic product-containing fluid from there and returning filtered aquatic product-containing fluid thereto; (ii) Multiple sensors capable of continuously measuring flow velocity, flow rate, and / or pressure at one or more locations along the filtration path and transmitting the measurement signals to a control device; and (iii) A connecting line capable of fluidly transferring filtered aquatic product-containing fluid to a downstream product outlet port; Equipped with, (c) One or more valved buffer inlets located within the outer edge of the cart, upstream of the first and second filtration paths, each buffer inlet being fluidically connected by a connecting line capable of fluidically transporting a desired aqueous fluid received from a reservoir or upstream unit operation for injection into the first or second filtration path by the operation of the first pump or the second pump and the valved buffer inlet in response to an instruction signal from the control device; and (d) The control device is configured and programmable to automatically switch the flow of the aquatic 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 and programmable to automatically switch the flow of the aquatic 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, wherein the control device receives the following instruction signals: (i) Activating a first pump during a desired continuous biological product production process, or, interchangeably, activating a second pump during a desired batch biological product production process; (ii) During a desired continuous biological product production process, a second pump is operated to allow buffer or water to flow through the first filtration path while the aquatic product-containing fluid flows through the second filtration path, or interchangeably, to allow buffer or water to flow through the second filtration path while the aquatic product-containing fluid flows through the first filtration path; and (iii) Controlling the valved product inlet and one or more valved buffer inlets, It can be further configured and programmed to issue [something].

[0074] Embodiment 2: The portable automatic single-use filter cart system of Embodiment 1, wherein the control device is configured and programmable for continuous production processes.

[0075] Embodiment 3: A portable automatic single-use filter cart system according to any of Embodiments 1 to 2, wherein the control device is configured and programmable for batch production processes.

[0076] Embodiment 4: A portable automatic single-use filter cart system according to any of Embodiments 1 to 3, wherein the control device is configured to interact with and programmable with a production equipment control system.

[0077] Embodiment 5: A portable automatic single-use filter cart system according to any of Embodiments 1 to 4, comprising a depth filter.

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

[0079] Embodiment 7: A portable, automated, single-use filter cart system according to any of Embodiments 1 to 6, comprising a sterile-grade filter.

[0080] Embodiment 8: A portable automatic single-use filter cart system according to any of Embodiments 1 to 7, comprising one or more portable accessory filtration modules.

[0081] The following embodiments are illustrative and should not be construed as limiting the scope of the present invention. [Examples]

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

[0083] [Process Setup] A continuous end-to-end (E2E) process from sub-perfusion to continuous capture was established, including a continuous two-tank virus inactivation filter followed by a continuous depth filter. A surge vessel was used between each unit operation to manage flow discrepancies between processing steps. Monoclonal antibody (mAb) products were perfused through a 0.2 μm filter, as previously described in Patent Document 1. Collection began on day 8 of production, and the downstream process was introduced when the surge vessel was filled to its processing volume.

[0084] 0.55m 2 A Millistak+ X0SP depth filter (Millipore Sigma) was used in conjunction with a Millipore Express SHC 0.2 μm sterile grade filter (Millipore Sigma) to remove turbidity and process-related impurities. The product load was set to last for 6 days of operation or until a maximum pressure of 20 psi was reached. The load could be set by k, volume, or time, and in this example, time-based switching was used. Production ended on day 22, and downstream operations were gradually stopped when the surge vessel filled to a minimum volume.

[0085] [Installation] As shown in Figures 1-3, a DF / VF tubing manifold (i.e., tubing system) equipped with probes, sensors, and a single-use pump head with AseptiQuik® fittings (CPC, Calder Products Company) was installed, and various parts of the tubing manifold were aseptically connected. The buffer connection line was aseptically attached to the inlet. The product load inlet was connected to a preceding supply surge container. To maintain a closed system, the discharged waste flow was connected together to a single waste outlet and connected to a drain pipe by a single-use air break assembly (see Patent Document 1). The product outlet port was connected to a recovery surge container. A depth filter was installed in a POD holder (catalog number MP0DPIL0T, Millipore Sigma), and lines from the "stage 1" (i.e., DF) position of the filter cart to the POD inlet and 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" position (i.e., sterile-grade filter) and aseptically connected to the tube manifold.

[0086] [Flushing (Cleaning)] The following steps were performed using an automated sequence in the DeltaV Distributed Control System (DCS) automation system (manufactured by Emerson).

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

[0088] Prime (fill with working fluid) the inlet connection line and depth filter to purge air from the system;

[0089] Rinse 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), and then up to the waste outlet connected thereto;

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

[0091] The NaOH is flushed out with equilibration buffer (EQ) first to the "Stage 1" waste outlet valve (410g), then to the final waste outlet valve (410j in Figure 4), and finally to the waste outlet connected thereto;

[0092] Allow automation to start loading the depth filter once the supply surge vessel reaches its starting weight. The depth filter load flow rate is based on the continuous process flow rate to maintain level control within the surge vessel.

[0093] The automation allowed the setpoint for the product load flow rate to be changed to high, normal, or low depending on the tank level in the preceding surge vessel, in order to control the volume in the surge vessel within a predetermined range. The initial buffer solution was kept in the connection line, the filter was cleaned and discharged through the waste outlet valve (410j in Figure 4), then switched to the product recovery outlet valve (410k in Figure 4), and then flowed to the product outlet port (403). The first filter was loaded for 6 days, and the automation monitored the system pressure while continuously controlling the flow rate.

[0094] [Filter Swap] Before the end of the load on filter path 1, the depth filter of filter path 2 (i.e., the second filtration path, arbitrarily represented as the lower path in Figure 4) was prepared in the same manner as all pre-use flushing performed on an automated filter cart system. Flushing of filter path 2 was performed simultaneously with the product load on filter path 1 using the valve configuration shown in Figure 4. When 10% of the load time for filter path 1 remained, the automation activated a flag to initiate the final EQ flush step for filter path 2. When filter path 1 reached the end of its load time, the automation switched the flow path of the product load from the load on filter path 1 to the load on filter path 2, and directed it towards the waste outlet valve (410j, lower path in Figure 4) of filter path 2 and to the waste outlet connected thereto. After the switch to loading the product onto filter path 2 occurred, automated EQ recovery and cleaning was initiated from the high-flow pump (416b) through filter path 1 to the product recovery outlet valve (410k, upper path in Figure 4) on filter path 1, and then to the product outlet port (403), recovering the product accumulated in the filter and piping connection lines. EQ recovery and cleaning on filter path 1 was performed simultaneously with the product loading on filter path 2. After reaching a predetermined volume transition point, automated switching of filter path 2 from the waste liquid valve (410j, lower path in Figure 4) to product recovery (410k, lower path in Figure 4) completed the operation of filter path 1. For the filter switch to the third set of depth filters, the same procedure was performed in the same manner as above using a new set of filters prepared on filter path 1, and once the loading of the second depth filter was complete, the system switched back to loading the product onto filter path 1. In this experiment, a total of three depth filters were used in this process, but the alternation between filter path 1 and filter path 2 can be continued for as many times as necessary for a given continuous process, if desired.

[0095] [result] [Flashing performance of depth filters (DF)] A flow rate of 2.3 liters / minute (L / min) was used for all washing steps. The pre-pressure of the depth filter was started at 10 psi and slowly reduced to 8-9 psi during washing. At the end of the analytical flush, the pH differed slightly between the filter path and the EQ buffer (0.1-0.3 pH units higher than the EQ buffer), which may be due to the pH probe offset after autoclaving. Standard procedure is to perform a one-point calibration adjustment on the system's pH transmitter to match the probe's pH with the offline measurement, but this procedure was not adopted in this test. Table 2 summarizes the washing 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 subjected to a flow load for a period of 6 days. As shown in Figure 5, a pressure increase was observed in each filter, and in filters 2 and 3, filter replacement occurred earlier than 6 days due to increased load turbidity. The pressure increase and load duration can be arbitrarily modified in future operations by increasing the area of ​​the filters used in the process.

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

[0099] [Process Setup] The process followed the DF process described in Example 1 (above), passing through two polishing chromatography steps prior to the virus filtering step before entering the surge vessel. The product was titrated with acid buffer in the preceding load surge vessel. Collection and recovery began on the 8th day of production, and the downstream process was introduced in stages when the surge vessel was filled to the working volume. In this process, 1000 L / m³ 2Planova (trademark) 20N virus removal filter (1m) 2 (Manufactured by Asahi Kasei) was used without replacement. Production ended on the 22nd day, and downstream operations were gradually stopped when the surge container filled to the minimum level.

[0100] [Installation] As described in Example 1 (above), a tube manifold equipped with a probe, sensor, and single-use pump head was installed, and various parts of the tube 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 the preceding supply surge container. The discharged waste flow was connected together to a single waste outlet to maintain a closed system, and this waste outlet was connected to a drain pipe by a single-use air break assembly (see Patent Document 1). The product outlet port was connected to the recovery surge container. The virus filter was installed in the "Stage 2" (VF) filter holder and aseptically connected to the inlet (supply) tube manifold and the outlet (permeate) tube manifold. The permeate was aseptically connected to a manually clamped tube. A 0.2 μm filter was inserted as a pre-filter at the "Stage 1" (DF) position and aseptically connected to the tube manifold.

[0101] [Pre-use leak test] In this demonstration run, a pre-use leak test was not performed. However, the system of the present invention is equipped with a dedicated valve that supplies air to pressurize the filter to the required level and to perform a pressure holding test according to the manufacturer's procedure.

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

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

[0104] Prime (fill with working fluid) the inlet connection line and depth filter to purge air from the system;

[0105] Wash the virus filter with water as recommended by the manufacturer, which includes purging air through the retaining liquid port; washing through the virus filter, the permeate flowing to the "Stage 2" (VF) waste outlet valve (410g, upper path in Figure 4), and from there to the waste outlet;

[0106] The equilibration buffer (EQ) buffer is passed through the permeate to the "Stage 2" waste outlet valve (410g, upper path in Figure 4), and then discharged into the waste outlet connected thereto;

[0107] Once the supply surge container reaches its starting weight and the supply tank reaches its pH setpoint, automation allows the loading of the virus filter to begin.

[0108] pH titration was performed in a supply surge vessel fitted with an acid buffer solution to a titration pump. Automation was used to achieve the pH setpoint. The automation temporarily suspended the virus filter load when the pH deviated from the processing specifications and resumed it when the pH returned to the range. The virus filter load flow rate was based on the continuous process flow rate to maintain level control within the surge vessel. The automation allowed the setpoint of the product load flow rate to be changed to high, normal, or low depending on the tank level in the preceding surge vessel in order to control the volume within the surge vessel within a predetermined range.

[0109] The initial buffer solution was retained in the connection line, the filter was cleaned and drained, and then the flow was switched to the product outlet port. Automation allowed for monitoring of the system pressure, and alarms alerted the operator in case of high pressure or flow rate deviations.

[0110] As described in Example 1, the virus filter can be replaced between filter path 1 (i.e., the first filtration path, optionally represented as the upper path in Figure 4) and filter path 2 (i.e., the second filtration path, optionally represented as the lower path in Figure 4), and back to filter path 1. Filter replacement can be performed for longer runs, but in this demonstration process, only one virus filter was used over the duration of the run, and no filter replacement was performed.

[0111] [result] The virus filter was subjected to continuous processing for 8 days. During this time, the inlet pressure of the Planova® filter showed a minimal increase from 0.7 to 1.3 psi (Figure 6). This application includes the following configuration . [Configuration 1] A portable automated single-use filter cart system for the production of replaceable, continuous or batch-type biological products, comprising: (a) Portable cart having an outer edge; (b) A valved product inlet located within the outer edge of the portable cart for receiving aquatic product-containing fluid from an upstream unit operation located outside the outer edge of the cart, wherein the upstream unit operation is fluidically connectable to the product inlet by a sealable connector and connecting line, the product inlet is connected to a first pump or interchangeably to a second pump via a connecting line capable of fluidly transporting the aquatic 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 the first and second pumps are configured to alternately pump the flow of the aquatic product-containing fluid to a first filtration path or a second filtration path; Here, the first filtration path and the second filtration path are, respectively: (i) a plurality of filters located downstream of the first pump, wherein the plurality of filters are fluidically connected in sequence along the filtration path; and optionally, one or more portable accessory filtration modules located outside the outer edge of the cart, which are fluidically connected to the filtration path and capable of receiving aquatic product-containing fluid from there and returning filtered aquatic product-containing fluid thereto; (ii) Multiple sensors capable of continuously measuring flow velocity, flow rate, and / or pressure at one or more locations along the filtration path and transmitting the measurement signals to a control device; and (iii) A connecting line capable of fluidly transporting filtered aquatic product-containing fluid to a downstream product outlet port; It is equipped with, (d) One or more valved buffer inlets located within the outer edge of the cart, upstream of the first and second filtration paths, each valved buffer inlet being fluidly connected by a connecting line capable of fluidly transporting a desired aqueous fluid received from a reservoir or upstream unit operation for injection into the first or second filtration path by the operation of the first or second pump and the valved buffer inlet in response to an instruction signal from the control device; and (e) The control device is configured and programmable to automatically switch the flow of the product-containing fluid from the first filtration path to the second filtration path in response to a predetermined measurement signal received from the sensor of the first filtration path, or after a predetermined period of time, wherein the control device is further configured and programmable to automatically switch the flow of the 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 of the second filtration path, wherein the control device receives the following instruction signals: (i) Operating the first pump during a desired continuous biological product production process, or interchangeably operating the second pump during a desired batch biological product production process; (ii) Operating the second pump during the desired continuous biological product production process to allow buffer or water to flow through the first filtration path while the aquatic product-containing fluid flows through the second filtration path, or interchangeably, allowing buffer or water to flow through the second filtration path while the aquatic product-containing fluid flows through the first filtration path; and (iii) Controlling the valved product inlet and the one or more valved buffer inlets, It is further configurable and programmable to issue Portable Automatic Single-Use Filter Cart System . [Configuration 2] The control device is configured and programmable for continuous production processes. Portable automatic single-use filter cart system as described in Configuration 1 above . [Configuration 3] The control device is configured for batch production processes. Portable automatic single-use filter cart system as described in any one of the above configurations 1 to 2 . [Structure 4] The control device is configured to interact with the production equipment control system and is programmable. Portable automatic single-use filter cart system as described in any one of the above configurations 1 to 3 。 [Composition 5] Equipped with a depth filter, Portable automatic single-use filter cart system as described in any one of the above configurations 1 to 4 。 [Composition 6] Equipped with a virus filter, Portable automatic single-use filter cart system as described in any one of the above configurations 1 to 5 。 [Composition 7] Equipped with a sterile-grade filter, Portable automatic single-use filter cart system as described in any one of the above configurations 1 to 6 。 [Structure 8] The aforementioned one or more portable accessory filtration modules are provided, Portable automatic single-use filter cart system as described in any one of the above configurations 1 to 7 。

Claims

1. A portable, automated, single-use filter cart system for the production of replaceable, continuous or batch-type biological products, comprising: (a) Portable cart having an outer edge; (b) A valved product inlet located within the outer edge of the portable cart for receiving aquatic product-containing fluid from an upstream unit operation located outside the outer edge of the cart, wherein the upstream unit operation is fluidically connectable to the product inlet by a sealable connector and connecting line, the product inlet is connected to a first pump or interchangeably to a second pump via a connecting line capable of fluidly transporting the aquatic 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 the first and second pumps are configured to alternately pump the flow of the aquatic product-containing fluid to a first filtration path or a second filtration path; Here, the first filtration path and the second filtration path are, respectively: (i) a plurality of filters located downstream of the first pump, wherein the plurality of filters are fluidly connected in sequence along the filtration path; and optionally, one or more portable accessory filtration modules located outside the outer edge of the cart, which are fluidly connected to the filtration path, capable of receiving aquatic product-containing fluid from there and returning filtered aquatic product-containing fluid thereto; (ii) Multiple sensors capable of continuously measuring flow velocity, flow rate, and / or pressure at one or more locations along the filtration path and transmitting the measurement signals to a control device; and (iii) A connecting line capable of fluidly transporting filtered aquatic product-containing fluid to a downstream product outlet port; It is equipped with, (d) One or more valved buffer inlets located within the outer edge of the cart, upstream of the first and second filtration paths, each valved buffer inlet being fluidly connected by a connecting line capable of fluidly transporting a desired aqueous fluid received from a reservoir or upstream unit operation for injection into the first or second filtration path by the operation of the first or second pump and the valved buffer inlet in response to an instruction signal from the control device; and (e) The control device is configured to automatically switch the flow of the product-containing fluid from the first filtration path to the second filtration path in response to a predetermined measurement signal received from the sensor 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 the 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 of the second filtration path, wherein the control device receives the following instruction signal: (i) Operating the first pump during a desired continuous biological product production process, or interchangeably operating the second pump during a desired batch biological product production process; (ii) Operating the second pump during the desired continuous biological product production process to allow buffer or water to flow through the first filtration path while the aquatic product-containing fluid flows through the second filtration path, or interchangeably, allowing buffer or water to flow through the second filtration path while the aquatic product-containing fluid flows through the first filtration path; and (iii) Controlling the valved product inlet and the one or more valved buffer inlets, It is further configured to output, Portable automatic single-use filter cart system.

2. The control device is configured for a continuous production process. The portable automatic single-use filter cart system according to claim 1.

3. The control device is configured for batch production processes. The portable automatic single-use filter cart system according to claim 1.

4. The control device is configured to interact with the production equipment control system. The portable automatic single-use filter cart system according to claim 1.

5. Equipped with a depth filter, The portable automatic single-use filter cart system according to claim 1.

6. Equipped with a virus filter, The portable automatic single-use filter cart system according to claim 1.

7. Equipped with a sterile-grade filter, The portable automatic single-use filter cart system according to claim 1.

8. The system includes one or more portable accessory filtration modules. The portable automatic single-use filter cart system according to claim 1.

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