Vertical-shaking bioreactor and high-throughput reactor set

Abstract

This invention provides a vertically oscillating bioreactor, comprising a reaction vessel for containing a reaction liquid, a magnetic rod placed within the reaction vessel, and a drive assembly for driving the magnetic rod to move up and down within the reaction vessel. The magnetic rod is made of magnetic or magnetically conductive material, and its density is less than or equal to the density of the reaction liquid within the reaction vessel. The drive assembly drives the magnetic rod to move. This vertically oscillating bioreactor, by incorporating a magnetic rod and a drive assembly, enables the magnetic rod to move up and down within the reaction vessel, overcoming the contradiction between stirring efficiency and shear force in traditional horizontal stirred reactors. It achieves gas-liquid mixing with extremely low shear force and promotes gas-liquid mass transfer through the disturbance of the gas-liquid interface by the magnetic rod.

Description

Technical Field

[0001] This invention relates to a vertical oscillating bioreactor and a high-throughput reactor assembly. Background Technology

[0002] The purpose of a bioreactor is to provide the optimal environment for the growth of cultured cells or the synthesis of products. These environmental parameters include, but are not limited to, dissolved oxygen (DO), partial pressure of carbon dioxide (pCO2), acidity (pH), osmotic pressure, ionic strength, substrate concentration, temperature, pressure, and shear stress. In traditional stirred tank reactors, including the roller flasks commonly used for laboratory-scale cell culture, these parameters are maintained constant and homogeneous through internal stirring. The stirrer is horizontal, parallel to the cross-section of the reaction vessel. This type of stirring easily creates vortices, affecting the stirring effect. To overcome this drawback, stirred tank reactors generally require baffles, but this results in high shear, causing significant damage to the cells. Although various stirrer designs aim to reduce shear forces, the problem cannot be fundamentally avoided as long as this horizontal stirring method is used. Simultaneously, to provide oxygen to the cells within the reactor, bioreactors generally require aeration. To improve oxygen transfer efficiency, microporous gas distributors are often used. However, research has found that the shear force generated when small bubbles break at the gas-liquid interface also causes significant damage to cells, even greater than the damage caused by stirring. To resolve these contradictions, wave-like bag-type bioreactors have been invented. This type of reactor spreads the culture medium over a large area, relying primarily on direct gas exchange at the gas-liquid interface, while mixing within the liquid phase occurs through the oscillation of the entire reaction bag. This design results in a very large footprint, making it unsuitable for use as a high-throughput parallel reactor.

[0003] In view of this, it is necessary to improve existing bioreactors to solve the above problems. Summary of the Invention

[0004] The purpose of this invention is to provide a vertically vibrating bioreactor to solve the problem of large shear forces caused by horizontal stirring in existing bioreactors.

[0005] To achieve the above objectives, the present invention provides a vertical oscillating bioreactor, comprising a reaction vessel for containing a reaction liquid, a magnetic rod placed inside the reaction vessel, and a drive assembly for driving the magnetic rod to move up and down inside the reaction vessel. The magnetic rod is made of magnetic material or magnetically conductive material, and the density of the magnetic rod is less than or equal to the density of the reaction liquid inside the reaction vessel. The drive assembly drives the magnetic rod to move.

[0006] As a further improvement of the present invention, the density of the magnetic rod is less than the density of the reaction liquid in the reaction vessel, and the end of the magnetic rod is provided with a groove or a hole.

[0007] As a further improvement of the present invention, the reaction vessel includes a receiving body having an opening and a cover for covering or opening the opening.

[0008] As a further improvement of the present invention, the cover is provided with an air inlet for supplying gas into the reaction vessel and an air outlet for discharging waste gas from the reaction vessel. The reaction vessel also includes an air inlet pipe protruding into the reaction vessel from the air inlet, and the air inlet and the air outlet are located at both ends of the cover.

[0009] As a further improvement of the present invention, the cover is provided with a sampling port, and the reaction vessel further includes a sampling tube protruding into the reaction vessel from the sampling port, with the sampling port located at one end near the cover.

[0010] As a further improvement of the present invention, the cover is provided with at least one feed port for replenishing the reaction vessel with liquid.

[0011] As a further improvement of the present invention, the driving component includes a magnetic element disposed outside the reaction vessel and a driving member for driving the magnetic element to move, wherein the driving member drives the magnetic element to move and thus drives the magnetic rod to move.

[0012] As a further improvement of the present invention, the magnetic component includes two magnetic rods disposed on both sides of the reaction vessel and a connector connecting the two magnetic rods, wherein the magnetic poles of the two magnetic rods are opposite, and the driving component includes a drive motor and a circumferential linear conversion device connecting the drive motor and the magnetic component.

[0013] As a further improvement of the present invention, the driving component includes a plurality of electromagnets disposed on both sides of the reaction vessel, and changing the magnetism of the plurality of electromagnets drives the magnetic rod to move.

[0014] As a further improvement of the present invention, the vertical oscillating bioreactor also includes a signal module and a dissolved oxygen sensor and a pH optical sensor electrically connected to the signal module and disposed in the reaction vessel.

[0015] As a further improvement of the present invention, the vertical oscillating bioreactor further includes a limiting shaft extending along the height direction, the magnetic rod is placed in the horizontal direction, and the magnetic rod has a through hole in the vertical direction for the limiting shaft to pass through.

[0016] The present invention also provides a high-throughput reactor assembly, which includes multiple vertically oscillating bioreactors as described above, wherein the reaction vessels of the multiple vertically oscillating bioreactors are arranged side by side.

[0017] As a further improvement of the present invention, the driving assembly includes two magnetic rods disposed on both sides of the reaction vessel, a connecting frame for connecting the magnetic rods of the plurality of vertical oscillating bioreactors, and the driving component includes a driving motor and a circumferential linear conversion device connecting the driving motor and the magnetic rods.

[0018] The beneficial effects of the present invention are: the high-throughput reactor group and the vertical oscillating bioreactor of the present invention, by setting up magnetic rods and driving components, can drive the magnetic rods to move up and down in the reaction vessel, breaking the contradiction between the stirring efficiency and shear force of the traditional horizontal stirring reactor, completing the gas-liquid mixing process with extremely low shear force, and promoting gas-liquid mass transfer by disturbing the gas-liquid interface with magnetic rods. Attached Figure Description

[0019] Figure 1 This is a schematic diagram of the structure of a high-throughput reactor group according to an embodiment of the present invention;

[0020] Figure 2 This is a schematic diagram of the structure of a high-throughput reactor group according to another embodiment of the present invention;

[0021] Figure 3 This is a schematic diagram of the structure of the vertical oscillating bioreactor of the present invention without the installation of a drive motor;

[0022] Figure 4 yes Figure 3 A schematic diagram of the structure of a vertical oscillating bioreactor from another direction;

[0023] Figure 5 This is a schematic diagram of the vertical oscillation bioreactor of the present invention;

[0024] Figure 6 This is a schematic diagram of the structure of a vertical oscillating bioreactor according to another embodiment of the present invention;

[0025] Figure 7 This is a schematic diagram of the structure of a vertical oscillation bioreactor according to another embodiment of the present invention;

[0026] Figure 8 This is a graph showing the change of the internal average liquid velocity over time during the start-up process of the vertical oscillating bioreactor of the present invention at 60 rpm;

[0027] Figure 9 This is a front view of the internal flow field of the vertical oscillating bioreactor of the present invention at 50 rpm;

[0028] Figure 10 This is a side view of the internal flow field of the vertical oscillating bioreactor of the present invention at 50 rpm. Detailed Implementation

[0029] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0030] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0031] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances. Furthermore, the technical features involved in the different embodiments of this invention described below can be combined with each other as long as they do not conflict with each other.

[0032] like Figures 1 to 7 As shown, the high-throughput reactor group 200 of the present invention includes a plurality of vertical oscillating bioreactors 100, and the reaction vessels 1 of the plurality of vertical oscillating bioreactors 100 are arranged side by side.

[0033] like Figures 3 to 7 As shown, the vertical oscillating bioreactor 100 includes a reaction container 1 for containing the reaction liquid, a magnetic rod 2 placed inside the reaction container 1, a drive assembly 3 for driving the magnetic rod 2 to move up and down inside the reaction container 1, a signal module 4, a dissolved oxygen sensor 5 and a pH optical sensor 6 electrically connected to the signal module 4 and disposed inside the reaction container 1, and a limiting shaft 8.

[0034] The reaction vessel 1 includes a containment body 11 with an opening, a cover 12 for covering or opening the opening, and a sampling tube 13. The containment body 11 and the cover 12 are made of non-magnetic or non-magnetic materials, preferably biocompatible materials. The containment body 11 is cuboid in shape, and its shape, through clever design, allows for high-throughput culture with adjustable throughput compared to the traditional cylindrical reaction vessel 1.

[0035] The cover 12 is provided with an air inlet 121 for supplying air into the reaction vessel 1, an air outlet 122 for discharging waste gas from the reaction vessel 1, a sampling port 123, and at least one feed port 124 for replenishing liquid into the reaction vessel 1.

[0036] The air inlet 121 is used to inject a mixture of air, oxygen, carbon dioxide, and nitrogen into the reaction vessel 1. Preferably, the introduced gas is a mixture of air and oxygen in any proportion, the flow rate and composition of which are automatically adjusted by the control system according to the oxygen demand of the cells cultured in the reaction vessel 1.

[0037] In one embodiment, the reaction vessel 1 further includes an air inlet pipe 14, which protrudes into the reaction vessel 1 from the air inlet 121 and extends further into the reaction liquid, thereby allowing air to be directly injected into the reaction liquid to improve solubility.

[0038] The air outlet 122 is used to discharge gas. In this embodiment, the air inlet 121 and the air outlet 122 are located at both ends of the cover 12, which can make the gas flow in a single and fixed direction, that is, form the convection of gas and liquid phases, and promote mass transfer.

[0039] In this embodiment, there are three feeding ports 124 to facilitate the replenishment of different materials or liquids.

[0040] The sampling tube 13 protrudes into the reaction vessel 1 from the sampling port 123. Its preferred position is on one side of the reaction vessel 1, with the sampling port 123 located near the end of the cover 12. This allows for greater space for the movement of the magnetic rod 2. The sampling port 123 protrudes into the reaction liquid to extract the liquid for use in line analysis and detection.

[0041] The dissolved oxygen sensor 5 and pH optical sensor 6 are in patch form and are used to detect dissolved oxygen and pH in the culture medium during the cultivation process. The oxygen sensor and pH optical sensor 6 are located at the bottom of the reaction vessel 1. In this embodiment, multiple reaction vessels 1 of the high-throughput reactor group 200 are provided with tracks 7 at their bottoms. The signal module 4 is located on the tracks 7, slides on the tracks 7, and can be electrically connected to the dissolved oxygen sensor 5 and pH optical sensor 6 of different vertical oscillating bioreactors 100.

[0042] The magnetic rod 2 is made of magnetic or magnetically conductive material. Specifically, the two ends of the magnetic rod 2 need to have opposite magnetic properties, rather than the entire magnetic rod 2 needing to be magnetic.

[0043] like Figure 6 As shown, the magnetic rod 2 is placed in a horizontal direction. In some embodiments, the magnetic rod 2 has a through hole in a vertical direction for the limiting shaft 8 to pass through, so as to prevent the magnetic rod 2 from deflecting when it moves up and down.

[0044] The density of the magnetic rod 2 is less than or equal to the density of the reaction liquid in the reaction vessel 1. Since the density of most of the reaction liquid is similar to that of water, the density of the magnetic rod 2 only needs to be less than or equal to the density of water.

[0045] When the density of the magnetic rod 2 is comparable to that of the reaction liquid, the weight of the magnetic rod 2 and the buoyancy of the reaction liquid on the magnetic rod 2 are roughly canceled out, and the magnetic rod 2 can be suspended in the reaction liquid. If it is necessary to control the up and down movement of the magnetic rod 2, it is only necessary to provide a magnetic field force that can overcome the drag force between the magnetic rod 2 and the reaction liquid.

[0046] When the density of the magnetic rod 2 is less than the density of the reaction liquid in the reaction vessel 1, a groove or hole can be provided at the end of the magnetic rod 2. The magnetic rod 2 will float on the liquid surface, and when the magnetic rod 2 moves up and down, some air can be introduced into the reaction liquid through the groove or hole, promoting gas-liquid mass transfer.

[0047] The driving component 3 drives the magnetic rod 2 to move, specifically by moving the magnetic rod 2 up and down. This linear motion stirring propels the liquid in the reaction vessel 1 to reciprocate in the vertical direction, achieving gas-liquid phase mixing with extremely low shear force and promoting mass transfer.

[0048] In one embodiment, the driving component 3 includes a magnetic element 31 disposed outside the reaction vessel 1 and a driving element 32 for driving the magnetic element 31 to move. The driving element 32 drives the magnetic element 31 to move, thereby driving the magnetic rod 2 to move.

[0049] The magnetic component 31 includes two magnetic rods 311 disposed on both sides of the reaction vessel 1 and a connector 312 connecting the two magnetic rods 311. The magnetic poles of the two magnetic rods 311 are opposite. The driving component 32 includes a drive motor 321 and a circular linear conversion device 322 connecting the drive motor 321 and the magnetic component 31. The drive motor 321 rotates, driving the circular linear conversion device 322 to convert the circular motion into a reciprocating linear motion, which in turn drives the magnetic rods 311 to move up and down. The magnetic rods 311 drive the magnetic rod 2 to move up and down, thereby improving the gas-liquid mixing effect.

[0050] Figure 8 The change in the average liquid velocity inside the vertical oscillating bioreactor 100 over time when it is started at a speed of 60 rpm is shown. It can be seen that the vertical oscillating bioreactor 100 reaches a pseudo-steady-state operation in about 2 seconds. Figure 9 and Figure 10 The internal liquid flow state of the vertical oscillating bioreactor 100 is shown when it is operating at 50 rpm. It can be seen that there are virtually no dead zones in the liquid flow.

[0051] like Figure 1 As shown, in the high-throughput reactor group 200, the magnetic rods 311 of the multiple vertical oscillating bioreactors 100 arranged side by side are fixedly connected by a connecting frame 313. Thus, only one drive motor 321 and one circumferential linear conversion device 322 are needed to achieve the effect of driving the magnetic rods 2 of the multiple vertical oscillating bioreactors 100 to move up and down. Furthermore, the magnetic poles of the magnetic rods 2 of two adjacent vertical oscillating bioreactors 100 are opposite, and the magnetic poles of the magnetic rods 311 of two adjacent vertical oscillating bioreactors 100 are also opposite.

[0052] like Figure 2 As shown, a drive motor 321 and multiple circular linear conversion devices 322 can also be provided. Each vertical oscillating bioreactor 100 is provided with a corresponding circular linear conversion device 322. The drive component 32 also includes a transmission rod 323 connecting the drive motor 321 and multiple circular linear conversion devices 322. One drive motor 321 drives multiple circular linear conversion devices 322 to work.

[0053] like Figure 7As shown, in another embodiment, the driving component 3 includes multiple electromagnets 33 disposed on both sides of the reaction vessel 1. Changing the magnetism of the multiple electromagnets 33 drives the magnetic rod 2 to move. Three electromagnets 33 are disposed on each side of the reaction vessel 1, and the corresponding electromagnets 33 on both sides have opposite magnetisms and opposite magnetic poles to the ends of adjacent magnetic rods 2. Energizing the upper and lower electromagnets 33 and changing the magnitude and direction of the current drives the magnetic rod 2 to move up and down. In this embodiment, the three electromagnets 33 on the same side have a phase difference of 120°. The two electromagnets 33 at the same horizontal height on both sides have a phase difference of 180°. At any given time, at least two electromagnets 33 on each side are applying a magnetic field to the magnetic rod 2, one repelling and the other attracting.

[0054] The high-throughput reactor assembly 200 and the vertical oscillating bioreactor 100 of this invention, by incorporating a magnetic rod 2 and a driving component 32, enable the magnetic rod 2 to move up and down within the reaction vessel 1. This overcomes the contradiction between stirring efficiency and shear force in traditional horizontal stirred reactors, achieving gas-liquid mixing with extremely low shear force. Furthermore, the magnetic rod 2's disturbance of the gas-liquid interface promotes gas-liquid mass transfer. This design is compact and can be combined into a high-throughput reactor assembly 200, with the number of vertical oscillating bioreactors 100 adjustable according to actual needs. This invention is applicable to mammalian cells, and particularly suitable for disposable cell culture reactors.

[0055] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0056] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A vertically oscillating bioreactor, characterized in that: The vertical oscillating bioreactor includes a reaction vessel for containing the reaction liquid, a magnetic rod placed inside the reaction vessel, and a drive assembly for driving the magnetic rod to move up and down within the reaction vessel. The magnetic rod is made of magnetic or magnetically conductive material, and its density is less than that of the reaction liquid inside the reaction vessel. The drive assembly drives the magnetic rod to move, and the magnetic rod floats above the reaction liquid as it moves up and down within the reaction vessel. The end of the magnetic rod has a groove or hole. The reaction vessel includes a receiving body with an opening for shielding... The cover is provided with an air inlet for supplying gas into the reaction vessel, an air outlet for discharging waste gas from the reaction vessel, and an air inlet pipe protruding into the reaction vessel from the air inlet. The air inlet and the air outlet are located at both ends of the cover. The cover is provided with a sampling port. The reaction vessel also includes a sampling pipe protruding into the reaction vessel from the sampling port. The sampling port is located at one end near the cover. The cover is provided with at least one feed port for replenishing liquid into the reaction vessel.

2. The vertical oscillating bioreactor according to claim 1, characterized in that: The driving assembly includes a magnetic component disposed outside the reaction vessel and a driving component for driving the magnetic component to move. The driving component drives the magnetic component to move, thereby driving the magnetic rod to move.

3. The vertical oscillating bioreactor according to claim 2, characterized in that: The magnetic component includes two magnetic rods disposed on both sides of the reaction vessel and a connector connecting the two magnetic rods. The magnetic poles of the two magnetic rods are opposite. The driving component includes a drive motor and a circumferential linear conversion device connecting the drive motor and the magnetic component.

4. The vertical oscillating bioreactor according to claim 1, characterized in that: The driving assembly includes multiple electromagnets disposed on both sides of the reaction vessel, and changing the magnetism of the multiple electromagnets drives the magnetic rod to move.

5. The vertical oscillating bioreactor according to claim 1, characterized in that: The vertical oscillating bioreactor also includes a signal module and a dissolved oxygen sensor and a pH optical sensor electrically connected to the signal module and disposed within the reaction vessel.

6. The vertical oscillating bioreactor according to claim 1, characterized in that: The vertical oscillating bioreactor also includes a limiting shaft extending along the height direction, the magnetic rod is placed in the horizontal direction, and the magnetic rod has a through hole in the vertical direction for the limiting shaft to pass through.

7. A high-throughput reactor assembly, characterized in that: The high-throughput reactor group includes a plurality of vertically oscillating bioreactors as described in any one of claims 2-6, wherein the reaction vessels of the plurality of vertically oscillating bioreactors are arranged side by side.

8. The high-throughput reactor array according to claim 7, characterized in that: The drive assembly includes two magnetic rods disposed on both sides of the reaction vessel and a connecting frame for the magnetic rods connecting multiple vertical oscillating bioreactors. The drive component includes a drive motor and a circumferential linear conversion device connecting the drive motor and the magnetic rods.

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