Microcarrier filter bag assembly and method of use

Abstract

The present invention relates to a filter bag assembly comprising a flexible bag delimiting a compartment configured to contain a fluid. An inlet port and an outlet port are each secured to the flexible bag so as to communicate with the compartment. A porous filter sheet is disposed within the compartment of the flexible bag such that fluid entering the compartment through the inlet port must pass through the filter sheet before exiting the compartment through the outlet port. A first retention seal secures the porous filter sheet to a portion of the flexible bag within the compartment, the first retention seal having an outer peripheral edge forming an annular continuous loop.

Description

[0001] This application is a divisional application of Chinese patent application number 201780073539.1 entitled “Microcarrier filter bag assembly and method of use” (which is an application that entered the national phase of international patent application PCT / US2017 / 063985). Technical Field

[0002] This invention relates to a filtration system and assembly for separating microcarriers from cell culture solutions. Background Technology

[0003] The biopharmaceutical industry commonly uses microcarriers to enable the growth of adherent cells. Specifically, microcarriers are used regularly during cell culture to optimize the growth of various adherent cell lines, such as protein-producing or virus-producing adherent cell populations, which are often used in the production of biologics (proteins) and vaccines.

[0004] Microcarriers possess surface chemistry that allows adherent-dependent cells to adhere to and grow during cell culture procedures. Microcarriers can be made from several different materials and typically have a density that allows them to be maintained in suspension with only gentle stirring.

[0005] Cell culture using microcarriers is typically performed in bioreactors. During culture, cells grow on the surface of the microcarriers. Once the cell culture process is complete, the cultured cells detach from the microcarriers through a chemical process performed in solution. The culture solution contains cells that are then separated from the microcarriers for use or further processing. The collected microcarriers can be cleaned, sterilized, and reused, or they can be discarded.

[0006] Typically, the separation of microcarriers from a culture solution containing detached cells is achieved by passing a solution through a rigid container with a horizontal sieve extending across the rigid container. The sieve is a rigid mesh that allows the culture fluid to pass through but prevents the microcarriers from doing so. However, when microcarriers accumulate on the sieve, they begin to clog it and prevent fluid from passing through. Once the sieve is clogged, the process stops until the sieve is clear. Furthermore, once the process is complete, the rigid container and associated sieve must be cleaned and sterilized before they can be reused. These process steps can be expensive and time-consuming.

[0007] Therefore, what is needed in this field is a method and / or system that can alleviate one or more of the above problems. Summary of the Invention

[0008] In a first independent aspect of the invention, a filter bag assembly includes:

[0009] The first flexible thin sheet;

[0010] A flexible second sheet is overlaid on and fastened to the first sheet to form a compartment therebetween;

[0011] At least one port is fastened to the first or second sheet to communicate with the compartment;

[0012] A porous filter sheet disposed between the first sheet and the second sheet, wherein the porous filter sheet is configured to filter fluid entering the compartment; and

[0013] A first retaining seal fastens the second sheet to the filter sheet such that at least a portion of the compartment surrounds the first retaining seal.

[0014] In one example, the filter sheet divides the compartment into a pre-filter compartment and a post-filter compartment, the pre-filter compartment surrounding the first retaining seal.

[0015] In another example, the outlet port is fastened to the first sheet or the second sheet, the at least one port including an inlet port communicating with the pre-filter compartment and an outlet portion communicating with the post-filter compartment.

[0016] In another example, a first sealing line secures the first sheet to the second sheet, and the retaining seal is spaced apart from the first sealing line.

[0017] In another example, the first sealing line forms a continuous loop.

[0018] In another example, a second sealing line is formed between the filter sheet and the second sheet, the second sealing line being in the form of a continuous loop surrounding the inlet port and the retaining seal, a first portion of the second sealing line including a portion of the first sealing line, and a second portion of the second sealing line being spaced apart from the first sealing line.

[0019] In another example, the filter bag assembly further includes:

[0020] A first sealing line, which secures the first sheet to the second sheet and forms a continuous loop; and

[0021] A second sealing line secures the filter sheet to the second sheet. The second sealing line is in the form of a continuous loop surrounding the inlet port and the retaining seal, and is spaced apart from the first sealing line.

[0022] In another instance, the first sheet and the second sheet comprise separate portions of a single, continuous sheet that has been folded up.

[0023] In one example, the porous filter sheet is arranged such that fluid entering the compartment through the inlet port must pass through the filter sheet before exiting the compartment through the outlet port.

[0024] In another example, the first retaining seal has an annular outer peripheral edge.

[0025] In another example, the first retaining seal is circular.

[0026] In another example, the first retaining seal has a ring configuration.

[0027] In another example, the first retaining seal has a maximum radius of at least or less than 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm or 15 cm from the center or center of the first retaining seal.

[0028] In another example, the first retaining seal has a minimum radius of at least 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm, or 15 cm from its center or center point.

[0029] In another example, a second retaining seal is spaced apart from the first retaining seal and fastens the second sheet to the filter sheet, with at least a portion of the compartment surrounding the second retaining seal.

[0030] In another instance, the first retaining seal also seals the first sheet to the filter sheet.

[0031] In another example, the filter sheet covers the inlet port but is spaced apart from the outlet port.

[0032] In another example, the first sheet and the second sheet each comprise a polymer film.

[0033] In another example, the first retaining seal causes the filter sheet to have multiple creases when the filter sheet is pushed away from the second sheet.

[0034] The first aspect of the invention may be included elsewhere herein, and may include any of the features, options, and possibilities set forth in other aspects of the invention.

[0035] In a second independent aspect of the invention, a filtration system comprises:

[0036] The support member is positioned at an angle between approximately 15° and 75° relative to the horizontal plane; and

[0037] The filter bag assembly according to claim 1 is mounted on a support tray.

[0038] In one example, a tube with a first end is connected to a bioreactor, and the opposite second end is coupled to the inlet port of the bag assembly.

[0039] The second aspect of the invention may be included elsewhere herein, including any of the features, options, and possibilities set forth in other aspects of the invention.

[0040] In a third independent aspect of the invention, a filter bag assembly comprises:

[0041] Flexible bags that limit the compartments configured to contain fluid;

[0042] An inlet port and an outlet port are each secured to the flexible bag to communicate with the compartment;

[0043] The porous filter sheet is disposed within the compartment of the flexible bag such that fluid entering the compartment through the inlet port must pass through the filter sheet before exiting the compartment through the outlet port; and

[0044] A first retaining seal secures the porous filter sheet to a portion of the flexible bag within the compartment, the first retaining seal having an outer peripheral edge forming an annular continuous loop.

[0045] In one instance, the outer peripheral edge of the first retaining seal is rounded.

[0046] In another example, the first retaining seal has a ring configuration.

[0047] In another example, the first retaining seal has a maximum radius of at least or less than 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm or 15 cm from the center or center of the first retaining seal.

[0048] In another example, the first retaining seal has a minimum radius of at least 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm, or 15 cm from its center or center point.

[0049] In another example, a second retaining seal is spaced apart from the first retaining seal and secures the porous filter sheet to a portion of the flexible bag.

[0050] In another example, the flexible bag includes a first flexible sheet and a second flexible sheet fastened together by a first sealing line surrounding the compartment, the first retaining seal being spaced apart from the first sealing line.

[0051] In another example, the porous filter sheet is secured to the second sheet by a second sealing line surrounding a portion of the porous filter sheet, the first retaining seal being spaced apart from the second sealing line.

[0052] In another instance, the first sealing line is spaced apart from the second sealing line, or at least a portion of the second sealing line forms part of the first sealing line.

[0053] In another example, the retaining seal is formed on the portion of the porous filter sheet surrounded by the second sealing line.

[0054] In another example, the first sheet and the second sheet each comprise a polymer film.

[0055] The third aspect of the invention may be included elsewhere herein, including any of the features, options, and possibilities set forth in other aspects of the invention.

[0056] In a fourth independent aspect of the invention, a method for filtering microcarriers from a liquid solution comprising cells comprises:

[0057] The liquid solution having the microcarriers is delivered into a compartment of a filter bag assembly, the filter bag assembly comprising:

[0058] A flexible bag that defines the compartment;

[0059] A porous filter sheet disposed within the compartment of the flexible bag; and

[0060] A first retaining seal, which secures a portion of the porous filter sheet to a portion of the flexible bag within the compartment; and

[0061] The liquid solution is passed through a porous filter sheet within the compartment of the flexible bag. The porous filter sheet is configured to prevent the microcarriers from passing through. When the microcarriers accumulate within the compartment of the filter bag, the filter bag expands to form multiple creases on the filter sheet.

[0062] In one instance, the crease protrudes radially outward from the first retaining seal.

[0063] In another example, the outer peripheral edge of the first retaining seal is rounded.

[0064] In another example, the first retaining seal has a ring configuration.

[0065] In another example, the first retaining seal has a maximum radius of at least or less than 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm or 15 cm from the center or center of the first retaining seal.

[0066] In another example, the first retaining seal has a minimum radius of at least 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm, or 15 cm from its center or center point.

[0067] In another example, a second retaining seal is spaced apart from the first retaining seal and secures the porous filter sheet to a portion of the flexible bag.

[0068] In another example, the flexible bag includes a first flexible sheet and a second flexible sheet fastened together by a first sealing line surrounding the compartment, the first retaining seal being spaced apart from the first sealing line.

[0069] In another example, the porous filter sheet is secured to the second sheet by a second sealing line surrounding a portion of the porous filter sheet, the first retaining seal being spaced apart from the second sealing line.

[0070] In another example, at least a portion of the compartment of the flexible bag surrounds the retaining seal.

[0071] The fourth aspect of the invention may be included elsewhere herein, and may include any of the features, options, and possibilities set forth in other aspects of the invention. Attached Figure Description

[0072] Various embodiments of the invention will now be discussed with reference to the accompanying drawings. It should be understood that these drawings merely depict typical embodiments of the invention and therefore should not be construed as limiting the scope of the invention.

[0073] Figure 1 This is a schematic representation of a bioreactor in which fluid is coupled to a filtration system;

[0074] Figure 2 yes Figure 1 A front perspective view of an embodiment of the filtering system shown in the image;

[0075] Figure 3 yes Figure 2 The image shows a front perspective view of the upright components of the filtration system.

[0076] Figure 4 yes Figure 2 The rear perspective view of the filtration system shown in the image;

[0077] Figure 5 yes Figure 2 The image shows a top plan view of the bag assembly of the filtration system.

[0078] Figure 6 yes Figure 5 The bottom plan view of the bag assembly shown in the image;

[0079] Figure 7 yes Figure 6 The exploded perspective view of the bag assembly shown in the image;

[0080] Figure 8 yes Figure 5 The cross-sectional side view of the bag assembly shown in the image;

[0081] Figure 9 yes Figure 5 The image shows a top plan view of the filter sheet and back plate of the bag assembly when it is empty.

[0082] Figure 10 This is in the case where the bag assembly is at least partially filled. Figure 9 The image shows a top view of the filter sheet and back panel.

[0083] Figure 10A It was cut along line 10A-10A. Figure 10 The image shows a cross-sectional view of the filter sheet.

[0084] Figure 11 It showcases a variety of alternative sealing elements formed on it. Figure 6 The bottom plan view of the bag assembly shown in the image;

[0085] Figure 12 yes Figure 8 A cross-sectional side view of an alternative embodiment of the bag assembly shown in the image;

[0086] Figure 13 It is used in conjunction with a relative structure to form a retaining seal. Figure 8 The cross-sectional side view of an alternative embodiment of the bag assembly shown in the figure; and

[0087] Figure 14 It has a relative structure in which the press bag is mechanically formed to maintain the seal. Figure 13 The image shows a cross-sectional side view of the bag assembly. Detailed Implementation

[0088] Before describing this disclosure in detail, it should be understood that this disclosure is not limited to the parameters of the specifically illustrated systems, methods, apparatuses, products, processes, compositions, and / or assemblies, which are of course variable. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments of this disclosure only and is not intended to limit the scope of this disclosure in any particular way. Therefore, although this disclosure will be described in detail with reference to specific embodiments, features, aspects, configurations, etc., the description is illustrative and should not be construed as limiting the scope of the claimed invention. Various modifications may be made to the illustrated embodiments, features, aspects, configurations, etc., without departing from the spirit and scope of the invention as defined by the claims. Therefore, while various aspects and embodiments have been disclosed herein, other aspects and embodiments are contemplated.

[0089] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While several methods and materials similar to or equivalent to those described herein may be used in the practice of this disclosure, only certain exemplary materials and methods are described herein.

[0090] Various aspects of the invention, including apparatus, systems, methods, etc., may be described with reference to one or more exemplary embodiments or implementations. As used herein, the terms “embodiment,” “alternative embodiment,” and / or “exemplary implementation” mean “serving as an example, illustration, or description” and should not be construed as being preferred or advantageous over other embodiments or implementations disclosed herein. Furthermore, references to “implementations” of this disclosure or the invention include specific references to one or more embodiments thereof, and vice versa, and such references are intended to provide illustrative examples without limiting the scope of the invention, the scope of which is indicated by the appended claims rather than the following description.

[0091] It should be noted that, unless the context clearly specifies otherwise, the singular forms “a” and “the” as used in this specification and the appended claims include multiple indicators. Thus, for example, reference to “retaining seal” includes one, two, or more retaining seals.

[0092] As used throughout this application, the word “may” is used in a permissive sense (i.e., meaning possible) rather than a mandatory sense (i.e., meaning must). Furthermore, the terms “including,” “having,” “involving,” “containing,” “characterized by,” and their variations (e.g., “includes,” “has,” “involves,” “contains,” etc.) and similar terms as used herein (including the claims) should be inclusive and / or open-ended, and should have the same meaning as the word “comprising” and its variations (e.g., “comprise” and “comprises”), and illustratively do not exclude additional undescribed elements or method steps.

[0093] Aspects of the invention can be illustrated by describing components that are coupled, attached, connected, and / or joined together. As used herein, the terms “coupled,” “attached,” “connected,” and / or “joined” are used to indicate a direct connection between two components, or, where appropriate, an indirect connection to each other via an intervening or intermediate component. In contrast, when a component is referred to as “directly coupled,” “directly attached,” “directly connected,” and / or “directly joined” to another component, no intervening element is present or anticipated. Therefore, as used herein, the terms “connected,” “connected,” etc., do not necessarily imply direct contact between two or more elements. Furthermore, components that are coupled, attached, connected, and / or joined together are not necessarily (reversibly or permanently) fastened to each other. For example, coupling, attaching, connecting, and / or joining may include placing, positioning, and / or arranging components together, or in some embodiments, otherwise placing, positioning, and / or arranging them adjacently.

[0094] As used herein, directional and / or arbitrary terms such as “top,” “bottom,” “front,” “back,” “left,” “right,” “up,” “down,” “above,” “below,” “inside,” “outside,” “inner,” “outer,” “inside,” “outer,” “inner,” “outer,” “near,” “far,” etc., are used only to indicate relative direction and / or orientation and are not intended to otherwise limit the scope of this disclosure, including this specification, the invention, and / or the claims.

[0095] Where possible, similar component designations have been used in the figures. Furthermore, similar components and / or multiple components with similar functions may be indicated by similar designations (e.g., component "10" and component "210"). Additionally, alternative configurations of specific components may each include a separate letter appended to the component designation. Accordingly, the appended letter may be used to indicate alternative designs, structures, functions, implementations, and / or embodiments of components or features that do not include the appended letter. Similarly, multiple instances of a component and / or its child components may each include a separate letter appended to the component designation. In each case, component designations may be used without the appended letter generally referring to any instance of the component or any alternative component. Component designations including appended letters may be used to refer to a specific instance of the component or to distinguish or attract attention to multiple uses of the component. However, component designations including appended letters are not intended to be limited to specific and / or particular embodiments described herein. In other words, references to specific features of one embodiment should not be construed as being limited to application only within the described embodiment.

[0096] It should also be understood that when a series of values ​​(e.g., less than, greater than, at least and / or up to a certain value, and / or between two stated values) are disclosed or described, so too are any specific values ​​or ranges of values ​​that are disclosed and anticipated to fall within the range of the disclosed values. Thus, a disclosure of an illustrative measurement or distance less than or equal to about 10 units or between 0 and 10 units illustratively includes the following specific disclosures: (i) measurements of 9 units, 5 units, 1 unit, or any other value between 0 and 10 units (inclusive of 0 units and / or 10 units); and / or (ii) measurements of any range of values ​​between 9 and 1 unit, between 8 and 2 units, between 6 and 4 units, and / or between 0 and 10 units.

[0097] It should also be noted that, according to certain embodiments of the present invention, systems, methods, apparatuses, devices, products, processes, compositions, and / or assemblies may include, incorporate, or otherwise include the properties, features, aspects, steps, components, parts, and / or elements described in other embodiments disclosed and / or described herein. Therefore, reference to specific features, aspects, steps, components, parts, elements, etc., with respect to one embodiment should not be construed as limiting application only to that embodiment. Furthermore, reference to specific benefits, advantages, problems, solutions, methods of use, etc., with respect to one embodiment should not be construed as limiting application only to that embodiment.

[0098] The headings used herein are for organizational purposes only and are not intended to limit the scope of the description or claims. For ease of understanding, similar reference numerals have been used where possible to denote similar elements common to the figures.

[0099] The present invention relates to various apparatuses and methods for effectively filtering microcarriers or other particles out of cell culture solutions while minimizing clogging or otherwise impeding the flow of the solution away from the microcarriers.

[0100] Figure 1 A cell culture system 10 incorporating the features of the present invention is depicted. In the cell culture system 10, cells are grown within a bioreactor 12, such as a bioreactor 12. The bioreactor 12 may be a microgravity bioreactor, an internally stirred bioreactor, a fluidized bed bioreactor, a rocker bag bioreactor, or any other type of bioreactor known in the art. The bioreactor 12 may also be a rigid tank bioreactor requiring sterilization between uses or a single-use bioreactor containing disposable bags. Alternatively, other types of bioreactors or other bioreactors, such as rotary flasks, may be used. Cells are grown in a nutrient growth medium that may contain a variety of different components. The components typically depend on the cell type and processing conditions. Growth media and related components are known in the art and are not discussed herein.

[0101] Microcarriers are added to the growth medium within bioreactor 12 to allow adherent-dependent cells to grow thereon. The microcarriers can be spherical beads, typically with a diameter between about 130 micrometers and about 300 micrometers. Other sizes may also be used. It should also be understood that the microcarriers may have alternative shapes, but typically have a maximum diameter that is typically at least or less than 130 micrometers, 170 micrometers, 200 micrometers, 250 micrometers, 300 micrometers, or within any two of the foregoing. The microcarriers have a density that allows suspension to be maintained by gentle agitation. For example, the microcarriers may also typically have a density of at least or less than 1.0 g / cm³. 3 1.02g / cm 3 1.05g / cm 3 1.10 g / cm 3 Or 1.20g / cm 3 Or a density within the range of any two of the foregoing items. Other densities are also possible. Microcarriers can be made from several different materials, including DEAE polydextrose, glass, polystyrene plastic, acrylamide, and collagen. Different types of microcarriers can differ in their porosity, specific gravity, optical properties, presence of animal components, and surface chemistry. Surface chemistry can include extracellular matrix proteins, recombinant proteins, peptides, and positively or negatively charged molecules. Microcarrier materials, along with different surface chemistry, can influence cell behavior, including morphology, proliferation, and adhesion.

[0102] During culture, cells grow on the surface of microcarriers disposed within the mixture. Once the cell culture process is complete, chemical reagents, such as enzymes, are added to the mixture, which contains growth medium, microcarriers suspended in the growth medium, and cells. The chemical reagents cause the cells to detach from the microcarriers, allowing the cells to suspend freely in the growth medium. The mixture is then removed from bioreactor 12 and passed through filtration system 14. As discussed in more detail below, filtration system 14 separates the microcarriers from the culture medium, which contains growth medium and detached cells. More specifically, the microcarriers may be captured by filtration system 14, while the culture medium may pass freely through filtration system 14. The culture medium may be returned to bioreactor 12 via line 16 or transported downstream via line 18 to another container or processing equipment for encapsulation or further processing.

[0103] like Figure 2 As depicted, the filtration system 14 includes a stand 20 that supports the filter bag assembly 22. Figure 3 and 4 As depicted, the stand 20 includes a tray 24, a support 26 erected on the tray 24, and a pair of arms 28A and 28B extending between the tray 24 and the support 26 to hold the support 26 at a desired angle relative to a horizontal plane. More specifically, the tray 24 has a bottom layer 30 with upright peripheral sidewalls 32 extending therefrom. The bottom layer 30 and the peripheral sidewalls 32 partially define the cavity 34. The tray 24 is elongated and extends between a first end 36 and an opposing second end 38. A retaining lip 40 is formed on and along the peripheral sidewalls 32 above the bottom layer 30. A pair of supports 42A and 42B are erected from the bottom layer 30 at the second end 38. Recesses 44A and 44B are formed on each support 42A and 42B.

[0104] The support member 26 is in the form of a panel having a front side 46 and an opposing back side 48, the front side extending between a first end 50 and an opposing second end 52, and between a first side 54 and an opposing second side 56. A pair of spaced-apart mounting holes 57A and 57B are recessed into or extend through the support member 26 at the first end 50. A first opening 58 passes centrally through the support member 26 at the first end 50, while a second opening 60 passes centrally through the support member 26 at the second end 52. A pair of recesses 62A and 62B are recessed into the back side 48 at or towards the first end 50 on the first side 54 and the second side 56, respectively.

[0105] During use, the second end 52 of the support 26 rests on the retaining lip 40 at the first end 36 of the tray 24. The arm 28A has a first end 64A received within a recess 44A in the cavity 34 and the bracket 42A, and an opposing second end 64B received within a recess 62A on the support 26. Similarly, the second arm 28B has a first end 66A received within a recess 44B in the cavity 34 and the bracket 42B, and an opposing second end 66B received within a recess 62B on the support 26. In this orientation, when the tray 24 is placed on a horizontal surface, the support 26 is positioned relative to the horizontal plane at an angle typically between 15° and 75°, with 25° and 65° or 35° and 55° being more common. In other embodiments, the angle α can be at least or less than 15°, 35°, 45°, 55°, 65°, 75°, or within any two of the foregoing. The angle α can be changed by using arms 28 of different lengths or by using arms 28 that are extended or retracted, for example. Thus, by extending the arm 28, the angle α increases, and by shortening the arm 28, the angle α decreases. When the stand 20 is not in use, the arm 28 can be removed from the tray 24 and the support 26 and stored within the cavity 34. The support 26 can also be lowered to rest at least partially within the cavity 34 on the lip 40.

[0106] like Figure 5 and 6 As depicted, the bag assembly 22 includes a bag 70 on which a first port 72A and a second port 72B are mounted. The bag 70 has a front side 76 and an opposing back side 78 extending between an upper end 80 and an opposing lower end 82, and also between a first side 84 and an opposing second side 86. A pair of spaced-apart attachment holes 88A and 88B extend at the upper end 80 through the bag 70 between sides 76 and 78.

[0107] like Figure 7 As depicted, the bag 70 consists of three sheets of material: a first sheet 90 forming a front side 76, a second sheet 92 forming a back side 78, and a filter sheet 94 sandwiched or otherwise disposed between the sheets 90 and 92. The first sheet 90 has an outer surface 100 and an opposing inner surface 101; the second sheet 92 has an outer surface 102 and an opposing inner surface 103; and the filter sheet 94 has a front side 104 and an opposing back side 105.

[0108] Sheets 90 and 92 comprise waterproof polymeric films, such as low-density polyethylene. The polymeric film may have a thickness of at least or less than 0.02 mm, 0.05 mm, 0.1 mm, 0.2 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, or within any two of the foregoing. Other thicknesses may also be used. The film is sufficiently flexible that it can be rolled into a tube without plastic deformation, and / or can be folded within angles of at least 90°, 180°, 270°, or 360° without plastic deformation.

[0109] The membrane may consist of a single layer of material or may comprise two or more layers that are sealed together or separated to form a double-walled container. When the layers are sealed together, the material may comprise a laminated or extruded material. A laminated material comprises two or more separately formed layers that are subsequently bonded together with an adhesive. Laminated and extruded films typically have between one and nine layers, and so on, typically between three and nine layers. The film used may typically have at least or fewer than one, three, five, seven, or nine layers, or several layers within the range of any two of the foregoing. The extruded film may be a cast film, such as a multilayer co-extruded cast film. An example of an extruded material that may be used in this invention is Thermo Scientific CX3-9 film, available from Thermo Fisher Scientific. The Thermo Scientific CX3-9 film is a three-layer, nine-mil cast film produced in a cGMP facility. The outer layer is a polyester elastomer co-extruded together with the ultra-low density polyethylene product contact layer. Another example of an extruded material that can be used in this invention is Thermo Scientific CX5-14 cast film, also available from Thermo Fisher Scientific. The Thermo Scientific CX5-14 cast film comprises a polyester elastomer outer layer, an ultra-low density polyethylene contact layer, and an EVOH barrier layer disposed therebetween.

[0110] The material can be approved for direct contact with living cells and is capable of maintaining the sterility of the solution. In this embodiment, the material can also be sterilized, for example, by ionizing radiation. Examples of materials that can be used in various situations are disclosed in U.S. Patent No. 6,083,587, published July 4, 2000, and U.S. Patent Publication No. US 2003-0077466 A1, published April 24, 2003, which are hereby specifically incorporated by reference.

[0111] like Figure 8As depicted, the bag 70 has an internal surface 96 that defines a compartment 98 between the first sheet 90 and the second sheet 92. The compartment 98 typically has a volume of at least or less than 1 liter, 3 liters, 6 liters, 10 liters, 15 liters, 20 liters, 30 liters, 50 liters, 75 liters, 100 liters, or within any two of the foregoing. Other volumes may also be used.

[0112] The filter sheet 94 comprises a material that allows the culture medium, i.e., growth medium and detached cells, to pass through while preventing microcarriers from passing through it. The filter sheet 94 may be composed of a porous material, such as a mesh, net, perforated sheet, porous sheet, lattice-type material, woven material, or any other material that allows the culture medium to pass through it while preventing associated microcarriers from passing through it. To allow cells to pass through the filter sheet 94 but prevent microcarriers from passing through it, the filter sheet 94 is typically made of a porous material with pores ranging in size from about 15 micrometers to about 100 micrometers, commonly from about 30 micrometers to about 100 micrometers. Where necessary, the filter sheet 94 may be expandable and / or elastically stretchable. Examples of materials that can be used for the filter sheet 94 include polyester (PET), polyamide (PA), polypropylene (PP), and polyetheretherketone (PEEK). Other materials may also be used, such as those discussed above for forming the first sheet 90 and the second sheet 92. It should also be understood that the filter sheet 94 may have a thickness of at least or less than 0.02 mm, 0.05 mm, 0.1 mm, 0.2 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, or within any two of the foregoing. Other thicknesses may also be used. The filter sheet 94 is generally flexible enough that it can be rolled into a tube without plastic deformation and can be folded at angles of at least 90°, 180°, 270°, or 360° without plastic deformation. The filter sheet 94 may be made of the same or different materials as sheets 90 and 92 and may have the same or different melting temperatures.

[0113] Each port 72 has a tubular guide rod 108, which has an annular flange 110 projecting radially outward from a first end and an annular wedge-shaped barb 112 formed on the opposite second end.

[0114] During one instance of the formation of bag assembly 22, such as Figure 7As depicted, holes 116A and 116B are centrally formed at their opposite ends on the second sheet 92. Valve stems 108 of ports 72A and 72B pass through holes 116A and 116B respectively from the inner surface 103 of the second sheet 92. A flange 110 is then fastened to the inner surface 103 of the second sheet 92, for example, by welding or using an adhesive. Next, the back surface 105 of the filter sheet 94 overlaps on top of the inner surface 103 of the second sheet 92, and the two sheets are partially fastened together. Specifically, in Figure 9 The image depicts a top plan view showing a filter sheet 94 that is covered and partially fastened to a second sheet 92.

[0115] In this embodiment, the filter sheet 94 is smaller than the second sheet 92 and is positioned such that the filter sheet 94 covers the first port 72A but not the second port 72B. That is, the filter sheet 94 has a bottom edge 122 disposed at a bottom end 123 spaced upwardly from the bottom edge 124 of the second sheet 92, such that the filter sheet 94 does not cover but is actually spaced from the second port 72B. The bottom edge 122 extends laterally to or extends toward the opposite side edge of the second sheet 92. A sealing line 121 is formed along the bottom end 123, for example along the bottom edge 122, to seal the filter sheet 94 to the second sheet 92 together. The sealing line 121 can be formed by applying thermal energy, radio frequency (RF) energy, acoustic energy, inductive energy, or other sealing energy to weld the filter sheet 94 to the second sheet 92 together, thereby forming a weld line. For example, when the second sheet 92 has a lower melting temperature than the filter sheet 94, energy can be applied to the overlying sheets 92 and 94 until a portion of the inner surface of the second sheet 92 melts and flows into and around the adjacent portion of the filter sheet 94. Once the energy is removed and the molten plastic cools and solidifies, the second sheet 92 and the filter sheet 94 are sealed / welded together. In other embodiments, the filter sheet 94 may have a lower melting temperature than the second sheet 92, such that the filter sheet 94 melts and bonds to the second sheet 92. Similarly, both the second sheet 92 and the filter sheet 94 may have the same or substantially the same melting temperature, such that when energy is applied, they both partially melt and bond to each other. In other embodiments, a polymeric material or other welding material may be placed on top of the filter sheet 94 or between the filter sheet 94 and the second sheet 92, such that when energy is applied, the polymeric material or other welding material partially or completely melts and seals / welds sheets 92 and 94, with or without directly melting sheets 92 and / or 94. In yet another embodiment, the sealing line 121 may be formed by applying an adhesive, a mechanical seal such as a crimping sleeve, or by using other conventional sealing techniques.

[0116] As will be discussed further below, retaining seal 130A also serves to secure filter sheet 94 to second sheet 92. Retaining seal 130A is spaced apart from sealing line 121 and is typically, though not essential, formed centrally on filter sheet 94. Retaining seal 130A can be formed using the same techniques as those discussed above for sealing line 121, namely welding, adhesives, mechanical fasteners, etc. In the depicted embodiment, retaining seal 130A is circular, and more specifically, in the form of a ring or annulus. When retaining seal 130A has a circular outer peripheral edge, retaining seal 130A will often have a radius that is at least or less than 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm, or 15 cm, or within any two of the foregoing. Other sizes may also be used. In alternative embodiments, it should be understood that retaining seal is not necessarily circular, but can have a variety of different configurations. For example, in Figure 11 In the embodiments depicted, retaining seal 130B is formed with an oval or elliptical configuration; retaining seal 130C is formed as a linear shape; retaining seal 130C is formed as a polygon, such as a triangle, square, rectangle, or other polygon with at least 5, 6, 7, 8, 9, or 10 sides; retaining seal 130E is formed with an irregular or asymmetrical peripheral edge 132; and retaining seal 130F is formed as a circle relative to the ring of retaining seal 130A. It should be understood that retaining seal 130 may also have other shapes.

[0117] When the retaining seal 130 has a non-circular outer perimeter, the retaining seal 130 may have a maximum radius extending from the center of the retaining seal 130 to the edge of the outer perimeter that is at least or less than 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm, or 15 cm, or within any two of the foregoing. Similarly, the retaining seal 130 may have a minimum radius extending from the center of the retaining seal 130 to the edge of the outer perimeter that is at least or less than 0.25 cm, 0.5 cm, 1 cm, 2 cm, 3 cm, 5 cm, 10 cm, or 15 cm, or within any two of the foregoing. Other dimensions may also be used. Figure 9 The embodiment shown also illustrates that the bag assembly 22 can be formed using a single retaining seal 130. However, in other embodiments, such as in Figure 11 In this embodiment, the bag assembly 22 may be formed of a plurality of spaced-apart retaining seals 130, for example, at least or less than two, three, four, five or six retaining seals 130. The plurality of retaining seals 130 may be of the same shape and / or size, or may be of different shapes and / or sizes.

[0118] Return to Figure 8 After the filter sheet 94 is fastened to the second sheet 92 at the sealing line 121 and retaining seal 130A, the inner surface 101 of the first sheet 90 overlaps the front surface 104 of the filter sheet 94 and is fastened to the combination of the filter sheet 94 and the second sheet 92. The first sheet 90 typically covers all the filter sheets 94 and covers the second port 72B. It is now formed as follows: Figure 5 and 8 The sealing line 136 shown generally extends around the periphery of the first sheet 90 and secures the first sheet 90 to the second sheet 92. When the filter sheet 94 is positioned between the first sheet 90 and the second sheet 92 along the sealing line 136, the filter sheet 94 is also sealed / welded to the sheets 90 and 92. The sealing line 136 can be formed using the same technique as the sealing line 121 discussed above. The sealing line 136 forms a continuous loop surrounding the first port 72A, the second port 72B, and the retaining seal 130. The sealing line 136 also covers and seals to the opposite ends of the sealing line 121. The sealing line 136 includes an upper sealing line portion 137 and a bottom sealing line portion 138. The upper sealing line portion 137 seals the sheets 90, 92 and 94 together, while the bottom sealing line portion 138 extends below the filter sheet 94 and directly seals the sheets 90 and 92 together, while also extending around the second port 72B.

[0119] The bottom sealing line portion 138 includes a base 140, which is generally C-shaped or U-shaped and partially curved around the second port 72B to form a reservoir 144 for collecting liquid around the second port 72B. The bottom sealing line portion 138 also includes a pair of arms 142A and 142B that slope downward from opposite sides of the bag 70 and connect to opposite sides of the base 140 to guide fluid to the reservoir 144 and the second port 72B.

[0120] Due to sealing line 136, such as Figure 8 The pre-filter compartment 126 shown is formed between the filter sheet 94 and the second sheet 92, wherein the sealing line 120 forms the outer edge of the pre-filter compartment 126.

[0121] In the above assembly configuration, compartment 98 is defined between sheets 90 and 92. Filter sheet 94 divides compartment 98 of bag 70 into pre-filter compartment 126 and post-filter compartment 128. Pre-filter compartment 126 is directly defined between filter sheet 94 and second sheet 92, and has an outer edge forming a continuous loop formed by the combination of upper sealing line portion 137 and sealing line 121. Figure 6As shown, sealing lines 137 and 121 combine to form sealing line 139. Thus, the pre-filter compartment 126 has a peripheral edge formed by the continuously surrounding sealing line 139, which secures the filter sheet 94 to the second sheet 92 and surrounds the first port 72A and holds the seal 130A but does not surround the second port 72B. It should be understood that the surrounding sealing line 139 need not be a circle, but can be any desired shape forming a continuous loop.

[0122] like Figure 8 As depicted, it should also be noted that in this configuration, the pre-filtration compartment 126 surrounds the retaining seal 130A. That is, due to the retaining seal 130A, the pre-filtration compartment 126 may have an annular or circular shape surrounding the retaining seal 130A. In other words, at least a portion of the compartment 98 surrounds the retaining seal 130A. The portion of the compartment 98 surrounding the retaining seal 130A may include the pre-filtration compartment 126. When two or more retaining seals 130 are formed, as previously discussed, the pre-filtration compartment 126 may surround each of the individual retaining seals 130. In the assembled configuration, as further detailed below, once the culture medium and microcarriers are delivered into the pre-filtration compartment 126 by means of the first port 72A, the microcarriers are captured within the pre-filtration compartment 126 because they are blocked by the sealing line 139 and cannot pass through the filter sheet 94. However, the culture medium can pass through the filter sheet 94.

[0123] The post-filtration compartment 128 includes the remaining portion of compartment 98 that does not include the pre-filtration compartment 126. More specifically, the post-filtration compartment 128 includes a region directly defined between the first sheet 90 and the filter sheet 94, and a region directly defined between the first sheet 90 and the second sheet 92, i.e., a region where the cultured solution can flow after it has passed through the filter sheet 94 from the pre-filtration compartment 126. Thus, during use, the culture medium and microcarriers enter the pre-filtration compartment 126 through the first port 72A. As the culture medium is transferred into the post-filtration compartment 128, the microcarriers are captured by the filter sheet 94 and held in the pre-filtration compartment 126. The culture medium then travels downward within the post-filtration compartment 128 and exits through the second port 72B.

[0124] As the microcarriers accumulate within the pre-filtration compartment 126, they push the filter slab 94 toward the first slab 90. In addition to maintaining the formation of the seal 130, the filter slab 94 is pushed against the first slab 90. In this configuration, the accumulated microcarriers will restrict or prevent the culture medium from flowing into the post-filtration compartment 128 and / or into the second port 72B. However, by forming the retaining seal 130, at least a portion of the filter slab 94 is retracted against the second slab 92 and away from the first slab 90, as... Figure 8As depicted, this allows the post-filtration compartment 128 to expand openly so that the culture medium can flow freely from the pre-filtration compartment 126 to the post-filtration compartment 128. Furthermore, one of the unique and unexpected benefits of some embodiments of the invention is that, by forming the retaining seal 130, multiple creases are formed in the filter sheet 94 as the microcarriers converge within the bag 70. The crease formation allows the culture medium to freely pass between the first sheet 90 and the filter sheet 94 and flow to the fluid channel of the second port 72B.

[0125] More specifically, Figure 9 This is a top plan view of the filter sheet 94 covering the second sheet 92 when bag 70 is empty. In this state, the filter sheet 94 is generally flat. In contrast, Figure 10 In the pre-filter compartment 126 of bag 70 ( Figure 8 The image shows a top plan view of a filter sheet 94 covering a second sheet 92, at least partially filled with microcarriers. In this state, the presence of a retaining seal 130A at the center, which secures the filter sheet 94 to the second sheet 92, causes multiple creases 150 to be formed on the filter sheet 94. The creases 150 typically project radially outward from the retaining seal 130A. However, the orientation of the creases 150 can vary depending on the configuration of the retaining seal 130A and the position and shape of other retaining seals 130 used concurrently. Furthermore, the creases 150 are typically formed only to approximately the height of the microcarriers gathered within the pre-filter compartment 126. Therefore, if the microcarriers are gathered at the level of the retaining seal 130A, the creases may be formed at and below the retaining seal 130A, but not above it. Figure 10A It more clearly shows the crease along 150. Figure 10 The cross-sectional view of the filter sheet 94 taken along section lines 10A-10A. Each crease 150 partially confines the fluid channel 152, allowing the culture medium to pass through it.

[0126] For example, refer to Figure 8 Even if the filter sheet 94 is stretched outward to contact the inner surface 101 of the first sheet 90 due to the aggregation of microcarriers, the culture medium in the post-filtration compartment 128 can still pass freely between the filter sheet 94 and the first sheet 90 by traveling along the fluid channels 152 formed by the creases 150. Therefore, the use of one or more retaining seals 130 can improve fluid flow through the bag 70, especially when microcarriers accumulate within the bag 70, thereby reducing handling and production time. The number, depth, orientation, and other properties of the creases 150 can vary based on the position, shape, and size of the retaining seals 130, the amount of microcarriers disposed within the pre-filtration compartment 126, and other variables.

[0127] The methods described above are merely examples of how bag assembly 22 can be manufactured. It should be understood that many other methods exist for forming bag assembly 22. By way of example, and not limitation, compared to forming the sealing line 121 and retaining seal 130A only when the second sheet 92 is applied over the filter sheet 94, as relative to... Figure 9 Describe and discuss, such as Figure 10 A sealing line 120, depicted as a continuous loop, may be formed between the filter sheet 94 and the second sheet 92. The sealing line 120 includes a sealing line 121 and forms the outer edge of the pre-filter compartment 126 between the filter sheet 94 and the second sheet 92, as previously discussed. Once the sealing line 120 is formed, the first sheet 90 may overlap the filter sheet 94, as previously discussed. (As previously discussed and...) Figure 5 The sealing line 136 shown can then be formed directly on top of the portion of the sealing line 120, so as to re-form as shown. Figure 8 The bag assembly 22 depicted in the text.

[0128] In another alternative, the filter sheet 94 may be smaller than the second sheet 92, such that some or all of the outer edges of the filter sheet 94 are spaced inwardly from the outer edges of the second sheet 92. Therefore, as... Figure 11 As depicted, the sealing line 120 formed between the filter sheet 94 and the second sheet 92 may be spaced inwardly from the sealing line 136 formed between the first sheet 90 and the second sheet 92. In this embodiment, no sealing line 136 may be overlaid or sealed to the filter sheet 94. The sealing line 120 and all other sealing lines discussed herein may be formed using the methods discussed above with respect to the sealing line 121.

[0129] In another embodiment, instead of the first sheet 90 and the second sheet 92 comprising two separate sheets, the first sheet 90 and the second sheet 92 may comprise a folded overlay of a single continuous sheet. The single sheet may be folded edge-to-edge or top-to-bottom, with the filter sheet 94 positioned between the overlapping portions. The same sealing lines and retaining seals may be formed as discussed above, except that the overlays of the sheets along the fold lines do not need to be sealed together, as they are integrally formed as a single continuous sheet.

[0130] like Figure 4 As depicted, the bag assembly 22 may further include a first fluid line 156A coupled to a first port 72A and a second fluid line 156B coupled to a second port 72B. Fluid lines 156A and 156B may include flexible tubing or other conduits. Once the bag assembly 22 is formed, it can be sterilized, for example, by radiation or other conventional sterilization techniques.

[0131] During use, component 20 is as follows Figure 3The bag 70 is depicted as standing upright and typically placed on a table or other supporting structure. The bag assembly 22A is then fastened to the front 46 of the support 26. Specifically, the free end of the first fluid line 156A passes through a first opening 58 on the support 26, while the free end of the second fluid line 156B passes through a second opening 60 on the support 26. The back 78 of the bag 70 then rests against the front 46 of the support 26, and the bag 70 is fastened to the support 26. In the depicted embodiment, the bag 70 can be positioned by making pins 158A and 158B (… Figure 2 ) through attachment holes 88A and 88B of bag 70 ( Figure 5 And enter the mounting holes 57A and 57B on the support member 26. Figure 3 To secure it to the support member 26. In other embodiments, it should be understood that a variety of different fasteners, clamps, hooks, hooks, etc., can be used to secure the bag assembly 22A / bag 70 to the support member 26.

[0132] In the attachment configuration, bag 70 is positioned at the same corresponding angle α as previously discussed relative to support 26. While bag assembly 22A / bag 70 can be placed horizontally or supported vertically during use, it has been found that holding bag assembly 22A / bag 70 at angle α during use results in improved fluid flow and thus reduced processing time. It should be understood that bag assembly 22A can be used without stand 20, and stand 20 can have various configurations to support and hold bag assembly 22A in the desired orientation, such as other stands, brackets, holders, hooks, etc.

[0133] Once the bag assembly 22A is secured to the stand 20, the free end of the first line 156A is fluidly coupled to the bioreactor 12, and the free end of the second line 156B is also fluidly coupled to the bioreactor 12 or to a separate container or other processing equipment. When separation of the microcarriers from the culture medium is desired, the combined microcarriers and culture medium are dispensed from the bioreactor 12 such that they travel through the first line 156A and through the first port 72A into the pre-filtration compartment 126 of the bag 70. As previously discussed, the microcarriers remain within the pre-filtration compartment 126 because they cannot pass through the filter sheet 94. However, the culture medium containing cells and nutrients travels through the filter sheet 94 into the post-filtration compartment 128 and then through the second port 72B out of the bag 70. The second line 156B then carries the culture medium back to the bioreactor 12 or to some other container or processing equipment. Once the microcarriers have gathered in the pre-filter compartment 126, the bag 70 can be used to transport the microcarriers for disposal, cleaning, or reuse.

[0134] In alternative embodiments, it should be understood that the bag assembly 22A can have a variety of different configurations. For example, in Figure 12A cross-sectional side view of an alternative embodiment of bag assembly 22B is depicted, wherein similar elements between bag assemblies 22A and 22B are identified by similar element symbols. Previously, with respect to bag assembly 22A, the first sheet 90 was directly welded to the second sheet 92 at the lower end 82 around the second port 72B. However, in contrast, in bag assembly 22B, the sealing line 136 for securing the first sheet 92 to the second sheet 92 completely covers the top of the sealing line 120, such that the filter sheet 94 is always positioned between the first sheet 90 and the second sheet 92. Also in this embodiment, with respect to securing to the second sheet 92, the second port 72B is secured to the first sheet 90 at the lower end 82.

[0135] Bag assembly 22B also includes a retaining seal 130A. However, unlike retaining seal 130A which is formed only between the second sheet 92 and the filter sheet 94, in bag assembly 22B, retaining seal 130A secures the first sheet 90, the second sheet 92, and the filter sheet 94 together. This can be achieved by simultaneously welding all three sheets together or by first welding two of the sheets (e.g., sheets 92 and 94) together and then subsequently welding the third sheet onto them. Similarly, retaining seals 130 of any desired size, shape, or number can be formed on bag assembly 22B.

[0136] In another alternative embodiment, instead of directly fastening ports 72A and 72B to the first sheet 90 and / or the second sheet 92, it should be understood that the filter sheet 94 may be fastened to the first sheet 90 and / or the second sheet 92. Ports 72A and / or 72B may then be fastened through holes 116 formed by the filter sheet 94 sealed together with the first sheet 90 or the second sheet 92. Ports 72A and 72B will then be fastened to the filter sheet 94.

[0137] Although bag assemblies 22A and 22B only show the use of one inlet port 72A and one outlet port 72B, in other embodiments, at least two, three or more inlet ports 72A may be formed on the bag assembly, and / or at least two, three or more outlets 72B may be formed on the bag assembly.

[0138] exist Figure 13 and 14Another alternative embodiment of bag assembly 22C is depicted, wherein similar elements between bag assembly 22A and bag assembly 22C are identified by similar element symbols. Bag assembly 22C may be the same as bag assembly 22A or 22B, except that, instead of forming retaining seal 130 by welding or using adhesive to fasten sheets 92 and 94 or combinations of sheets 90, 92 and 94 together to form retaining seal 130, retaining seal 130 may be formed by mechanically fastening sheets 90, 92 and 94 together. For example, a first structure 166 and a second structure 168 may be placed on opposite sides of bag assembly 22C. In use, structures 166 and 168 may be pressed together such that portions of sheets 90, 92 and 94 are mechanically clamped and held together to form retaining seal 130G. When no longer in use, structures 166 and 168 can be separated to allow the sheets 90, 92, and 94 to be freely separated, thereby removing the retaining seal 130G. In one embodiment, structure 168 may consist only of support 26.

[0139] The embodiments of the present invention offer several unique benefits. For example, the bag assembly 22 is inexpensive to manufacture and can be discarded after a single use, eliminating the need for cleaning. Furthermore, the bag assembly achieves high fluid flow rates, even when it becomes filled with microcarriers, thereby reducing processing time and increasing efficiency. Other benefits are also achieved.

[0140] The invention may be implemented in other specific forms without departing from its spirit or essential characteristics. The described embodiments should be considered illustrative rather than restrictive in all respects. Therefore, the scope of the invention is indicated by the appended claims rather than by the foregoing description. All variations within the meaning and scope of equivalents of the claims are covered within the scope of the claims.

Claims

1. A filter bag assembly, comprising: The first flexible thin sheet; A flexible second sheet is overlaid on and fastened to the first sheet so that a compartment is formed between the second sheet and the first sheet; A porous filter sheet is disposed between the first sheet and the second sheet to divide the compartment into a first compartment and a second compartment. The porous filter sheet is made of an elastically stretchable material to prevent multiple microcarriers from passing through it. The first port is fastened to the second sheet so as to communicate directly with the first compartment; The second port is fastened to the second sheet to communicate directly with the second compartment, and the second port is spaced apart from the first port; The porous filter sheet is configured such that fluid entering the first compartment through the first port must pass through the porous filter sheet before entering the second compartment or exiting through the second port. The porous filter sheet is applied to the first port, but spaced apart from the second port and not applied to the second port.

2. The filter bag assembly according to claim 1, characterized in that, The second sheet, the first sheet, and the porous filter sheet are all sheets that cover each other.

3. The filter bag assembly according to claim 1, characterized in that, The filter bag assembly further includes: A first sealing line secures the first sheet to the second sheet and forms a continuous loop, the continuous loop surrounding the first port and the second port; and A second sealing line secures a portion of the porous filter sheet to the second sheet at a location between the first port and the second port.

4. The filter bag assembly according to claim 3, characterized in that, The second sealing line is linear and extends between two spaced-apart portions of the first sealing line.

5. The filter bag assembly according to claim 3, characterized in that, The second sealing line does not directly seal with the first sheet.

6. The filter bag assembly according to claim 3, characterized in that, The first sealing line directly seals a portion of the porous filter sheet to the first sheet and the second sheet.

7. The filter bag assembly according to claim 3, characterized in that, The second sealing line and a portion of the first sealing line combine to surround and demarcate the first compartment.

8. The filter bag assembly according to claim 1, characterized in that, The first sheet and the second sheet each have a first end and a opposite second end. The first ends of the first sheet and the second sheet are connected together, and the second ends of the first sheet and the second sheet are connected together. The first port is located at the first end of the second sheet, and the second port is located at the second end of the second sheet.

9. The filter bag assembly according to claim 8, characterized in that, The porous filter sheet is disposed at the first end of the first sheet and the second sheet, between the first sheet and the second sheet, but not at the second end of the first sheet and the second sheet.

10. The filter bag assembly according to claim 1, characterized in that, The porous filter sheet includes a single sheet having an inner surface in direct communication with the first compartment and an opposing outer surface in direct communication with the second compartment.

11. The filter bag assembly according to claim 1, characterized in that, The first sheet and the second sheet each comprise individual portions of a single, continuous sheet that has been folded up.

12. The filter bag assembly according to claim 1, characterized in that, It also includes multiple microcarriers for growing cells collected in the first compartment.

13. A method for filtering a suspension using a filter bag assembly according to claim 1, the suspension comprising a liquid and microcarriers suspended in the liquid, the method comprising: The suspension is delivered into the first compartment through the first port, the liquid passes through the porous filter sheet and enters the second compartment and exits through the second port, while the microcarriers are collected in the first compartment through the porous filter sheet; as well as Collect the liquid flowing out of the second port.

14. The method according to claim 13, characterized in that, It also includes connecting the first port of the filter bag assembly to the bioreactor containing the suspension prior to the delivery step.

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