Small bioreactor

Abstract

A small bioreactor comprising a cap, sidewalls, and a base, each of which can generally be manufactured as a single integral component. The bioreactor is sterilizable and configured to perform a bioprocess for the production of therapeutic agents (e.g., cell culture) and is configured to be disposed of after use. The base may further include an integrally formed spray line and impeller shaft, thereby creating an easily customizable and manufacturable bioreactor.

Description

Technology Field

[0001] The present invention relates to a bioreactor system and a method for operating the same, and more specifically, to a disposable small bioreactor and a method for using the same. Background Technology

[0002] Various vessels, devices, components, and unit operations for performing biochemical and / or biological processes or manipulating liquids and other products of such processes are known. To avoid the time, cost, and difficulties associated with sterilizing vessels used in biopharmaceutical manufacturing processes, single-use or disposable bioreactors and single-use mixers are used as such vessels. For example, biological materials (e.g., animal and plant cells) including mammalian, plant, or insect cells and microbial cultures can be processed using disposable or single-use mixers and bioreactors.

[0003] Generally, small bioreactors used during process development are designed so that impellers, spargers, and probes are inserted into the bioreactor through the top, for example, through a top cap. While this may provide a simpler bioreactor vessel, it makes scaling the desired process more difficult. For instance, when larger bioreactors implement bottom-driven impellers, using a small top-driven impeller negatively impacts scalability. This is attributed, at least in part, to the difference in mixing characteristics between top-driven and bottom-driven impellers. Furthermore, inserting spray tubes and probes through the top cap creates arbitrary baffling locations that negatively affect flow patterns, power levels, and overall mixing performance, and deviates further from the typical geometry of larger bioreactors.

[0004] However, biopharmaceutical manufacturers desire the ability to scale their processes from laboratory to production without significant changes. This minimizes additional costs, enabling clients to retain the value of their initial development and ensuring the same performance is achieved at a commercial scale. This helps clients minimize the cost of bringing their products to market.

[0005] Therefore, there is a need for small bioreactors that mimic the entire geometric structure of large-scale bioreactors, making scaling up simpler, more efficient, and cost-effective.

[0006] A first aspect of the present invention relates to a bioreactor (100) comprising a cap (101); a side wall (120) connected to the cap (101); and a base (150) connected to the side wall (120) through a sealing mechanism (105), wherein the cap (101), the side wall (120), and the base (150) define an internal volume (121), and the base (105) defines a bottom surface (154) that defines a fluid-containing portion (122) configured to hold a cell culture; an integral impeller shaft (165); and at least one integral spray line (131, 132), and the bioreactor is configured to perform cell culture at a maximum working volume of 2L or less.

[0007] In an embodiment, an integrated impeller shaft (165) and at least one spray line (131, 132) are integrated toward the central portion of the bottom surface (154). At least one spray tube (131, 132) and the impeller shaft (165) are each formed integrally with the base (150). The bioreactor (100) further comprises an impeller (164) connected to the impeller shaft (165), the impeller (164) comprises at least one magnet (167), and the impeller (164) is configured to rotate by the application of a magnetic field outside the bioreactor.

[0008] In an embodiment, the base (150) further comprises at least one port (102, 103, 107, 151, 158, 162) integrally formed inside, and the at least one port (102, 103, 107, 151, 158, 162) is configured so that a probe can pass through it into the internal volume. The base (150) further comprises a base lip (159) and at least one support leg (158). The magnetic drive unit (300) is configured to magnetically couple to the impeller (164) when the base lip (159) contacts the housing (301) of the magnetic drive unit (300).

[0009] In an embodiment, the base lip (159) includes at least one first attachment mechanism (156, 156', 160, 160') configured to be attached to at least one second attachment mechanism (302, 302') on the magnetic drive unit (300) so that the bioreactor (100) is physically connected to the magnetic drive unit (300).

[0010] In an embodiment, the bioreactor (100) is sterilizable and configured to perform cell culture for the production of a therapeutic agent, and the bioreactor (100) is configured to be discarded after performing cell culture.

[0011] A second aspect of the present invention relates to a method for culturing cells, the method comprising the step of providing a bioreactor (100) comprising a cap (101), a side wall (120) connected to the cap (101), and a base (150) connected to the side wall (120) through a sealing mechanism (105). The cap (101), the side wall (120), and the base (150) define an internal volume (121). The base comprises a bottom surface (154) defining a fluid-containing portion (122) configured to hold a cell culture; an integral impeller shaft (165); and at least one integral spray line (131, 132) for culturing cells within the bioreactor (100), wherein the cultivation is performed with a working volume of 250 ml to 2 L.

[0012] In an embodiment, an integrated impeller shaft (154) and at least one spray line (131, 132) are integrated toward the central portion of the bottom surface (154). At least one spray tube (131, 132) and an impeller shaft (165) are each formed integrally with the base (150). An impeller (164) is connected to the impeller shaft (165), and the impeller (164) includes at least one magnet (167). This method further includes the step of rotating the impeller (164) by applying a magnetic field outside the bioreactor (100).

[0013] In an embodiment, the base (150) further comprises at least one port (102, 103, 107, 151, 158, 162) integrally formed inside, and the method further comprises the step of passing a probe through at least one port (102, 103, 107, 151, 158, 162) in a bioreactor so that the probe is at least partially within the internal volume (121).

[0014] In an embodiment, the base (150) further comprises a base lip (159) and at least one support leg (158). The magnetic drive unit (300) is configured to magnetically couple to an impeller (164) when the base lip (159) contacts the housing (301) of the magnetic drive unit (300). The base lip (159) includes at least one first attachment mechanism (156, 156', 160, 160') configured to attach to at least one second attachment mechanism (302, 302') on the magnetic drive unit (300) so that the bioreactor (100) is physically connected to the magnetic drive unit (300).

[0015] In an embodiment, the method further includes the step of sterilizing the bioreactor (100) before the step of culturing cells, and the step of discarding the bioreactor (100) after the step of culturing cells is completed, and the cell culture is for producing a therapeutic agent. Brief explanation of the drawing

[0016] The present invention will be better understood by reading the following description of non-limiting embodiments with reference to the accompanying drawings, wherein: FIG. 1 is a perspective view of a small bioreactor according to an embodiment of the present invention. FIG. 2 is a cross-sectional view of the small bioreactor of FIG. 1 attached to a base according to an embodiment of the present invention. FIG. 3 is a perspective view of the base portion of the bioreactor shown in FIG. 1 and FIG. 2 according to an embodiment of the present invention. FIG. 4 is a cross-sectional view of a portion of the base of FIG. 3 according to an embodiment of the present invention. FIG. 5 illustrates the base of the bioreactor shown in FIG. 1 and FIG. 2 according to an embodiment of the present invention. FIG. 6 illustrates a bioreactor according to an additional embodiment of the present invention. Specific details for implementing the invention

[0017] Exemplary embodiments of the present invention will be described in detail below and are illustrated in the accompanying drawings. Where possible, the same reference numerals used throughout the drawings refer to the same or similar parts.

[0018] As used herein, an element or step described in the singular and used with "a" or "an" should be understood as not excluding a plurality of said elements or steps unless such exclusion is explicitly stated. Furthermore, references to "one embodiment" are not intended to be interpreted as excluding the existence of additional embodiments that also include the cited feature. Also, unless explicitly stated otherwise, an embodiment that "includes" or "has" an element or a plurality of elements having a specific characteristic may include additional such elements that do not have that characteristic.

[0019] It should be noted that the arrangement of components (e.g., number, type, arrangement, etc.) of the illustrated embodiment may be modified in various alternative embodiments. For example, in various embodiments, different numbers of probe ports, tube barbs, as well as the volume of the space intended for mixing may be modified, and this is within the scope of the present invention.

[0020] As used herein, a structure, restriction, or element "configured" to perform a task or operation may be formed, configured, or adapted particularly structurally in a manner corresponding to the task or operation. For clarity and to avoid doubt, an object that can be merely modified to perform a task or operation is not "configured" to perform a task or operation as used herein. Instead, the use of "configured" as used herein indicates a structural adaptation or characteristic and indicates the structural requirements of any structure, restriction, or element described as "configured" to perform a task or operation.

[0021] An embodiment of the present invention provides a bioprocessing system (10), in particular a small bioreactor system (10). In one embodiment, the bioprocessing system (10) comprises a bioreactor (100) that defines an internal volume (121), and the vessel (100) generally comprises three parts, namely a cap (101), a side wall (120), and a base (150). When joined together, the cap (101), the side wall (120), and the base (150) define an internal volume (121), which is suitable for performing bioprocesses (e.g., cell culture, cell proliferation, and mixing).

[0022] Referring to FIGS. 1 and 2, the bioreactor (100) comprises a side wall (120) that is sealed to a cap (101) at one end and to a base (150) at the opposite end through a sealing mechanism (105), thereby forming a fluid and gas-tight seal between the cap (101) and the side wall (120) and between the base (150) and the side wall (120). The sealing mechanism (105) may include a tongue and groove connection sealed by an O-ring or gasket. Alternatively, the sealing mechanism (105) may include a snap-fit ​​component, a screw fastener, and a magnet or chemical adhesive. Additionally, the sealing mechanism (105) may be formed by welding the cap (101) and the base (150) to the side wall (120), for example, through heat sealing, ultrasonic welding, etc. These connections between the cap (101) and the side wall (120) and between the base (150) and the side wall (120) form an internal volume (121) that is sealed from external air and contamination. Optionally, one or more baffles (123) are disposed extending downward along the length of the side wall (120) in the direction of the base (150).

[0023] The cap (101) includes a top wall (104) and a side (107). As illustrated in FIGS. 1 and 2, at least one hose bar (102, 103, 106) is connected to the cap (101) or formed integrally with the cap. In the embodiment illustrated in FIG. 1, at least one hose bar (102, 103) is located on the top wall (104), while at least one hose bar (106) is located on the side (107). These hose bar (102, 103, 106) may have any diameter or length and may be bilateral structures protruding into the internal volume (121) and away from the cap (101). The number of hose bar (102, 103, 106) and their respective positions on the cap (101) may be changed and are within the scope of the invention. The hose barb (102, 103, 106) can be implemented for various purposes such as liquid addition, liquid removal, exhaust, probe connection, etc.

[0024] The cap (101) has a generally circular / cylindrical shape, as best illustrated in FIG. 1, and the length of the side (107) is equal to the total height of the cap (101). Although not explicitly illustrated, the cap (101) may take other shapes, such as a square, hexagon, parallelogram, etc.

[0025] In a preferred embodiment, the cap (101) is a single-piece structure and can be manufactured through known methods (e.g., injection molding, 3D printing, etc.). Manufacturing complexity is reduced by integrating the parts. Additionally, so that the cap (101) is configured for single use and can be disposed of after use, the cap (101) can be manufactured from a polymer, more specifically, a sterilizable plastic. In an alternative embodiment, the cap (101) can be manufactured from a plurality of components.

[0026] Similar to the cap (101), the side wall (120) generally has a cylindrical shape, and its upper portion is configured to fit into the cap (101) as shown in FIGS. 1 and 2. As discussed in detail above, the side wall (120) is sealed to the side (107) of the cap (101) to form a fluid-tight seal. The side wall (120) may be manufactured from the same or similar material as the cap (101). Also, similar to the cap (101), the side wall (120) may take other shapes such as a square, a hexagon, a parallelogram, etc., provided that the shape is complementary to the cap (101).

[0027] As discussed above, the base (150) is connected to or otherwise sealed to the side wall (120) to define an internal volume (121) facing the cap (101). The base (150) is sealed to the side wall (120) in the same or similar manner as the cap (101) is sealed to the side wall (120) (i.e., by implementing a sealing mechanism (105)), the details of which are omitted here for brevity. The base (150) includes an inclined bottom surface (154) (e.g., a concave surface) to create a fluid containment section (122), the fluid containment section being the lower part of the internal volume (121) and accommodating an impeller shaft (165), an impeller (164), and a spray disc (133, 134), the details of which will be described in more detail below. In an alternative embodiment, the bottom surface (154) does not need to be sloped (for example, it may have a side wall and a bottom surface that meet to form a right angle).

[0028] Referring to FIGS. 3 and 4, components of the base (150) are illustrated. According to an embodiment, spray lines (131, 132) are integrally formed with the base (150) and terminate at the bottom surface (154). These spray lines (131, 132) allow fluidic access to the fluid-containing portion (122) and the internal volume (121), thereby enabling gas to be introduced into the internal volume (121) during a bioprocess operation (e.g., cell culture). As best seen in FIG. 4, spray discs (133, 134) are located at the ends of the spray lines (131, 132) and are also integrated into the bottom surface (154) of the base (150). In a preferred embodiment, the spray disc (133, 134) is a porous disc that ensures that the gas introduced into the internal volume (121) forms bubbles when it comes into contact with a liquid (e.g., cell culture medium). In a preferred embodiment, the spray line (131, 132) is circular at the distal end and has a first inner diameter, and the inner diameter increases when terminated at the bottom surface (154) (e.g., taking a conical shape), but other shapes and geometric structures are also within the scope of the invention. At opposite ends, the spray line (131, 132) is terminated at a connection (135, 136) (e.g., a hose bar) that can be attached to a source of ambient air or other gas intended to be introduced into the internal volume (121).

[0029] The impeller (164) and the impeller shaft (165) are generally located in the center of the bottom surface (154) of the base (150). In a preferred embodiment, the drive shaft (165) is formed integrally with the base (150), as best illustrated in FIG. 4. The impeller (164) is configured to rotate around the impeller shaft (165) to agitate the fluid located within the internal volume (121). In one embodiment, agitation occurs through magnetic coupling between a magnet (167) located within the impeller (164) and a magnet within the magnetic drive unit (300). The spray lines (131, 132) and spray discs (133, 134) are configured to be positioned below the impeller (164) (e.g., toward the center of the base (150)), so that when gas is introduced into the internal volume (121), the impeller (164) disperses the gas into the liquid within the internal volume (121).

[0030] By integrally forming the spray lines (133, 134) and the impeller shaft (165) on the base (150), the manufacturing of the bioreactor (100) is simplified. Additionally, the integral configuration of the base components allows for easy modification of the design of the impeller and sprayer. For example, this integral configuration allows the base (150) to be 3D printed, thereby allowing for easy changes to the diameter of the spray lines (131, 132), the number of spray lines (131, 132), and / or the size of the impeller (164) without significantly altering the manufacturing process (e.g., without the need to create a new mold).

[0031] However, it should be noted that the present invention is not explicitly limited to integrally formed spray lines (133, 134) and impeller shaft (165). Rather, the spray lines (133, 134) and impeller shaft (165) may be attached to the base (150) or welded in other ways. Even without these components being integrally formed within the base, the bioreactor can still be manufactured more easily (e.g., compared to a top drive system).

[0032] As further illustrated in FIG. 3, there are one or more hose barbs (151, 157, 162) protruding from the base (150) and extending away from the internal volume (121). The hose barbs (151, 157, 162) provide fluid access to the internal volume (121) and the fluid-containing portion (122). In an embodiment, the hose barbs (151 and 162) are sized so that a measurement probe can be attached and passed through for measuring cell cultures. Examples of such measurements include pH, DO, and temperature. In an embodiment, the hose barb (158) is sized for other purposes such as sampling, feeding, perfusion, harvesting / discharge, etc.

[0033] In an embodiment, the support leg (158) extends from the side of the base (150). The support leg (158) can be concentrically spaced around the base (150) and separated by a space (161) that facilitates access to the bottom surface of the base (150), thereby providing a space where the connection to the spray line (131, 132) can be operated and the base (150) can be aligned with the magnetic drive unit (300) shown in FIGS. 2 and 5.

[0034] The support leg (158) terminates at the base lip (159). The base lip (159) is the bottom surface of the base (150) and is larger in circumference than the rest of the base (150) so that the base lip (159) provides structural support for the bioreactor (100). The base lip (159) is paired with the housing (301) of the magnetic drive unit (300) as shown in FIGS. 2 and 5. In an embodiment, this pairing is achieved through complementary fastening mechanisms (156, 160, 302) on the base (150) and the housing (301). In one example shown in FIGS. 2 and 5, the housing (301) includes at least one dowel (302) configured to be fitted into at least one slot (156, 160) on the base lip (159). Alternative embodiments for pairing the base (150) and the magnetic drive unit (300) include, but are not limited to, snap fits, pressure pins, chemical or magnetic bonding, or clamps. When paired, the drive head (304) of the magnetic drive unit (300) is magnetically coupled to a magnet (167) located within the impeller (164). In an alternative embodiment, the impeller shaft passes through the base (150) and can be driven using a motor shaft.

[0035] The embodiments illustrated in FIGS. 1 through 4 illustrate a bioreactor having a first maximum working volume (e.g., 2 liters). According to additional embodiments, the bioreactor may have a smaller (or larger) working volume. For example, FIG. 6 illustrates a bioreactor (200) having a second maximum working volume smaller than the first working volume (e.g., 250 ml). The bioreactor (200) includes the same features as the bioreactor (100) but on a smaller scale, and a description of said features will be omitted to avoid repetition. The magnetic drive unit (300) includes a second set of dowels (302') located on the housing (301) closer to the central drive head (304). The dowels (302') are configured to align with at least one corresponding slot (156', 160') on the base lip (159)'. In this way, a single magnetic drive unit (300) can support bioreactors (100, 200) of various sizes.

[0036] As described above, the bioreactor (100, 200) is composed of three main components, namely a cap, a sidewall, and a base, each of which can generally be manufactured as a single component. The bioreactor (100, 200) is sterilizable and configured to perform a bioprocess (e.g., cell culture) for the production of therapeutic agents and is configured to be disposed of after use.

[0037] In this way, a small, single-use, floor-driven bioreactor (100, 200) (i.e., an impeller located at the bottom of the internal volume and driven by a floor-mounted motor) can be manufactured easily and robustly. Since these floor-driven bioreactors have the same overall geometry and mixing characteristics, they can be more easily scaled up to larger-scale floor-driven bioreactors.

[0038] Furthermore, as previously mentioned, bioreactor components can be manufactured via 3D printing, which enables easy and robust customization without requiring complex manufacturing modifications (e.g., no need to create new molds). In other words, by integrally forming the spray line, spray disc, impeller shaft, and / or hose barb with other components (i.e., cap and base), design changes can be easily made without costly modifications to the molding design. In this way, the bioreactor is highly flexible, scales well into larger floor-driven bioreactors, and is easily manufactured.

Claims

Claim 1 A bioreactor (100) comprises: a cap (101); a side wall (120) connected to the cap (101); and a base (150) connected to the side wall (120) through a sealing mechanism (105), wherein the cap (101), the side wall (120), and the base (150) define an internal volume (121), and the base (150) comprises, A bottom surface (154) defining a fluid-containing portion (122) configured to hold a cell culture; Integrated impeller shaft (165); and A bioreactor comprising at least one integrated spray line (131, 132), wherein the bioreactor is configured to perform cell culture with a maximum working volume of 2L or less. Claim 2 A bioreactor according to claim 1, wherein the integrated impeller shaft (165) and at least one spray line (131, 132) are integrated toward the central portion of the bottom surface (154). Claim 3 In paragraph 2, the bioreactor, wherein at least one spray tube (131, 132) and an impeller shaft (165) are each formed integrally with the base (150). Claim 4 A bioreactor according to claim 1, further comprising an impeller (164) connected to the impeller shaft (165), wherein the impeller (164) comprises at least one magnet (167). Claim 5 In paragraph 4, the bioreactor is configured such that the impeller (164) rotates by the application of a magnetic field outside the bioreactor (100). Claim 6 A bioreactor according to claim 1, wherein the base (150) further comprises at least one port (102, 103, 106, 151, 157, 162) integrally formed inside. Claim 7 In claim 6, the bioreactor is configured such that at least one port (102, 103, 106, 151, 157, 162) allows a probe to pass through it into the internal volume. Claim 8 A bioreactor according to claim 1, wherein the base (150) further comprises a base lip (159) and at least one support leg (158). Claim 9 In claim 8, the magnetic drive unit (300) is configured to magnetically couple to the impeller (164) when the base lip (159) comes into contact with the housing (301) of the magnetic drive unit (300), in a bioreactor. Claim 10 In claim 9, the base lip (159) comprises at least one first attachment mechanism (156, 156', 160, 160') configured to be attached to at least one second attachment mechanism (302, 302') on the magnetic drive unit (300) so as to physically connect the bioreactor (100) to the magnetic drive unit (300). Claim 11 In claim 1, the bioreactor (100) is sterilizable and configured to perform cell culture for the production of a therapeutic agent, and the bioreactor (100) is configured to be discarded after performing cell culture. Claim 12 As a method for culturing cells, a step of providing a bioreactor (100) wherein the bioreactor is, Cap (101); Side wall (120) connected to the above cap (101); and It includes a base (150) connected to the side wall (120) through a sealing mechanism (105), and The above cap (101), side wall (120) and base (150) define an internal volume (121), and The above base is, A bottom surface (154) defining a fluid-containing portion (122) configured to hold a cell culture; Integrated impeller shaft (165); and A method comprising at least one integral spray line (131, 132), a step of culturing cells in the bioreactor (100), wherein the cultivation is performed with a working volume of 250 mL to 2 L. Claim 13 In claim 12, the method wherein the integrated impeller shaft (165) and at least one spray line (131, 132) are integrated toward the central portion of the bottom surface (154). Claim 14 In claim 13, the method wherein at least one spray tube (131, 132) and an impeller shaft (165) are each formed integrally with the base (150). Claim 15 In claim 12, the impeller (164) is connected to the impeller shaft (165), and the impeller (164) includes at least one magnet (167). Claim 16 In claim 12, a method of rotating the impeller (164) by applying a magnetic field outside the bioreactor (100). Claim 17 In claim 12, the method further comprises at least one port (102, 103, 107, 151, 158, 162) integrally formed within the base (150). Claim 18 A method according to claim 17, wherein the probe is passed through at least one port (102, 103, 107, 151, 158, 162) in the bioreactor so that the probe is at least partially within the internal volume (121). Claim 19 In claim 12, the method further comprises the base (150) including a base lip (159) and at least one support leg (158). Claim 20 A method according to claim 12, wherein the magnetic drive unit (300) is configured to magnetically engage with the impeller (164) when the base lip (159) comes into contact with the housing (301) of the magnetic drive unit (300). Claim 21 A method according to claim 20, wherein the base lip (159) comprises at least one first attachment mechanism (156, 156', 160, 160') configured to be attached to at least one second attachment mechanism (302, 302') on the magnetic drive unit (300) so as to physically connect the bioreactor (100) to the magnetic drive unit (300). Claim 22 A method according to claim 20, further comprising the step of sterilizing the bioreactor (100) before the step of culturing the cells, and the step of discarding the bioreactor (100) after the step of culturing is completed, wherein the cell culture is for producing a therapeutic agent.

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