Sterile sampler and sterile sampling fixed bed bioreactor

Abstract

The utility model relates to the technical field of bioreactors, and discloses a sterile sampler and a sterile sampling fixed bed bioreactor, the bioreactor comprises a tank body, a stirring bin and a culture bin used for installing a fixed bed, the tank body is provided with an upper cover, and the upper cover is provided with a pipeline structure used for being communicated with the interior of the tank body; a consumable replacement opening is formed in the upper cover, a top cover is detachably and hermetically connected to the consumable replacement opening of the upper cover, and the culture bin is connected to the inner side of the top cover; the stirring bin is connected to one side, far away from the top cover, of the culture bin and is suspended in the tank body; a sampling opening is formed in the top cover, and a sterile sampler is arranged at the sampling opening. The consumables and the tank body are relatively independent, the consumables can be mounted without opening the upper cover of the tank body, and a tank body pipeline cannot be influenced; and the stirring device adopts a disposable structure, so that the reduction of foreign matters generated by abrasion is facilitated.

Description

Technical Field

[0001] The present application relates to the technical field of bioreactors, and in particular to a sterile sampler and a fixed-bed bioreactor for sterile sampling. Background Art

[0002] Fixed-bed bioreactors utilize non-cytotoxic, cell-adhesive materials as cell culture carriers. Cells grow by attaching to the culture carriers rather than suspending in culture medium. In fixed-bed bioreactors, cells and the growth carrier serve as the stationary phase, while the culture medium serves as the mobile phase. Through the continuous circulation of the culture medium, nutrients, metabolic waste, and gases are exchanged between the cells and the culture medium. Fixed-bed bioreactors provide a gentle, stable 3D culture environment for adherent cell culture, simulating the in vivo cell growth environment to the greatest extent possible, allowing cells to grow more efficiently and supporting the continuous collection of target products.

[0003] In the prior art, bioreactors generally use a suspended agitator tank, in which a filler (usually a sheet carrier) is fixed between two fixed plates as a stationary phase. The pretreatment is cumbersome and has large anisotropy, resulting in uneven cell growth and inability to perform cell sampling operations. Another commonly used technology is microcarriers, which also have the problems of high cost, cumbersome pretreatment procedures, and large-scale cell shedding and microcarrier breakage in the late stage of culture, especially after infection. As a result, even if sampling and counting are possible, the counting results are not very accurate or not very representative. Utility Model Content

[0004] In order to improve the convenience of consumables installation, the present application provides a sterile sampler and a fixed-bed bioreactor for sterile sampling.

[0005] The present application provides a fixed-bed bioreactor for sterile sampling, which adopts the following technical solution:

[0006] A sterile sampler comprises a hose, one end of which is sealedly connected to a sampling port, and the other end is sealedly connected to an upper plug; a lower plug is provided in the hose between the sampling port and the upper plug, and a slit is provided on the lower plug; the end of a sampling strip provided on a fixed bed passes through the sampling port and is provided in the hose, and the sampling strip clamp is provided in the slit; a tube clamp is provided on the hose.

[0007] By adopting the above technical solution, when sampling, the tube clamp is opened, the upper plug is removed, and the sampling strip is pulled out of the tank body using a tool. When the sampling strip is pulled to a certain position, the required sampling strip is located at a section of the lower plug close to the tank body; then the hose is cut using a shearing tool, and then the lower plug is sealed on the end of the hose still connected to the tank body, and the hose is clamped with a tube clamp to ensure the sterility of the tank body; the removed sampling strip is wrapped in the hose for subsequent test data analysis.

[0008] A fixed-bed bioreactor for sterile sampling comprises a tank body, a stirring chamber and a culture chamber for installing a fixed bed, the tank body being provided with an upper cover, the upper cover being provided with a pipeline structure for connecting to the interior of the tank body; the upper cover being provided with a consumables replacement port, the upper cover being detachably and sealingly connected to a top cover at the consumables replacement port, the culture chamber being connected to the inner side of the top cover; the stirring chamber being arranged in the tank body; the top cover being provided with a sampling port, the sampling port being provided with a sterile sampler.

[0009] By adopting this technical solution, the consumables are relatively independent from the tank body. Consumables can be installed without opening the tank cover, which will not affect the tank piping, making installation more convenient. They are also easy to disassemble and replace, reducing the number of steps and time required. Therefore, the rational piping structure and consumable replacement port design facilitate user maintenance and consumable replacement of the reactor.

[0010] Optionally, the stirring chamber is connected to a side of the culture chamber away from the top cover and is suspended in the tank body.

[0011] By adopting the above technical solution, the stirring device and the culture chamber are integrated into a disposable product, a disposable stirring structure, which reduces the amount of foreign matter caused by wear and tear. Most stirring devices on the market use polytetrafluoroethylene stirrers, which will wear out after long-term operation, producing black particles or colloids that affect cell growth. Furthermore, the stirring device is fixed to the bottom of the tank, requiring regular inspection and maintenance during use and is not easy to replace.

[0012] Optionally, the culture chamber includes an inner cylinder, which is connected to the top cover, a bracket is provided in the inner cylinder, a fixed bed is provided on the bracket, a first reflux hole is provided on the side wall of the inner cylinder close to the top cover, and the bottom of the inner cylinder is connected to the stirring chamber.

[0013] By adopting the above technical solution, the inner cylinder is connected to the top cover to realize the suspension setting of the culture chamber; and the fixed bed is supported by the bracket, and the first reflux hole is set so as not to affect the flow of the cell culture medium during the stirring process.

[0014] Optionally, the bracket includes a central column and at least two limiting rods fixedly connected to the central column. The fixed bed is centered on the central column and is limited between the limiting rods and the inner cylinder. The central column and the top cover are fixedly connected.

[0015] By adopting the above technical solution, the stability of the fixed bed is improved by fixing the central column and the top cover.

[0016] Optionally, the stirring bin includes a bin body, the bin body is fixedly connected to an end of the inner cylinder away from the upper cover, a stirring rod is provided in the bin body, and a second reflux hole is provided at the bottom of the bin body.

[0017] By adopting the above technical solution, the tank body is suspended, the stirring device and the culture tank are integrated as a whole, and it is a disposable product. The disposable stirring structure not only helps to reduce foreign matter caused by wear, but also facilitates removal and is not easy to affect the pipeline layout in the tank body.

[0018] Optionally, a glass cover is fixedly connected to the bottom of the tank body, the second reflux hole is provided on the glass cover, and a set gap is reserved between the glass cover and the bottom of the tank body.

[0019] By adopting this technical solution, the glass surface is smooth, and the magnetic stirring bar can form a relatively uniform vortex when rotating on the glass, thus achieving a better stirring effect. Compared with stainless steel, the surface of stainless steel may have tiny bumps and impurities, which will affect the stirring effect of the magnetic stirring bar when rotating on stainless steel. In addition, the smooth glass surface is easy to clean, making it easy to remove any residue that may be generated by the magnetic stirring bar during the stirring process.

[0020] Optionally, a sealing member is sealed between the consumable replacement port and the top cover.

[0021] By adopting the above technical solution, the sealing member helps to improve the sealing performance of the connection between the top cover and the consumable replacement port.

[0022] Optionally, the top cover and the upper cover are detachably connected and fixed by a plurality of hand-tightened bolts.

[0023] By adopting the above technical solution, manual disassembly and installation are facilitated by manually tightening the bolts.

[0024] Optionally, the inner cylinder and the top cover are connected via a plurality of fixing members.

[0025] By adopting the above technical solution, multiple fixing parts are used to facilitate the removal, replacement and installation of the fixed bed.

[0026] In summary, this application includes at least one of the following beneficial technical effects:

[0027] 1. The new structure makes the consumables and the tank relatively independent. The consumables can be installed without opening the tank cover, which will not affect the tank pipeline and is more convenient to install.

[0028] 2. The PTFE stirring bars currently on the market will wear out after long-term operation, producing black particles or colloids, which affect cell growth. The new structure combines the stirring chamber and the culture chamber into one, which can be used as a disposable product, effectively solving the adverse effects caused by wear.

[0029] 3. The new structure can realize the sterile sampling function of the fixed-bed bioreactor; cell culture can be carried out in an environment other than the biosafety cabinet. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a schematic diagram of the overall structure of a fixed-bed bioreactor for sterile sampling in an embodiment of the present application.

[0031] Figure 2 This is a schematic diagram of an explosion of a fixed-bed bioreactor for aseptic sampling in an embodiment of the present application.

[0032] Figure 3 This is a schematic diagram of the overall structure of an integrated stirring chamber and a culture chamber in a fixed-bed bioreactor for sterile sampling in an embodiment of the present application.

[0033] Figure 4 This is a cross-sectional view of a fixed-bed bioreactor for aseptic sampling in an embodiment of the present application.

[0034] Figure 5 This is a structural diagram of a fixed-bed bioreactor for sterile sampling in an embodiment of the present application, mainly used to display the support.

[0035] Figure 6 yes Figure 2 Cross-sectional view after explosion.

[0036] Figure 7 This is a schematic diagram of the bottom structure of the integrated stirring chamber and culture chamber in an embodiment of the present application.

[0037] Figure 8 This is a schematic diagram used to illustrate the second structure of the sterile sampling device in the embodiment of the present application.

[0038] Figure 9 This is a cross-sectional view of the hose after sampling in an embodiment of the present application.

[0039] Figure numerals: 1. tank body; 2. upper cover; 3. pipeline structure; 4. consumables replacement port; 5. top cover; 6. sampling port; 7. inner cylinder; 8. first reflux hole; 9. center column; 10. limit rod; 11. fixed bed; 12. bin body; 13. stirring rod; 14. second reflux hole; 15. glass cover; 16. sealing member; 17. hand-tightening bolt; 18. fixing member; 19. hose; 20. pipe clamp; 21. sampling strip; 22. mounting groove; 23. threaded hole; 24. mounting hole; 25. fixing hole; 26. wrap-around hole; 27. pipeline joint; 28. upper plug; 29. ​​lower plug. DETAILED DESCRIPTION

[0040] Embodiments of the present application are described in detail below, examples of which are illustrated in the accompanying drawings.

[0041] Throughout this specification, reference to the terms "certain embodiments," "one embodiment," "some embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0042] The present application discloses a fixed bed bioreactor for aseptic sampling, referring to Figure 1 and Figure 2 and Figure 3 , including a tank body 1, a stirring chamber and a culture chamber for installing a fixed bed 11. In this embodiment, the tank body 1 is a cylindrical stainless steel tank with a size of 2m. A circular upper cover 2 is installed on the tank body 1, and the outer peripheral side of the upper cover 2 is detachably connected to the tank mouth of the tank body 1 through a plurality of bolts distributed in a circumferential array. A pipeline structure 3 for connecting the interior of the tank body 1 is provided on the upper cover 2, and the pipeline structure 3 includes a pipeline connector 27 and a pipeline, and a sensor including a detection end extending into the tank body 1. A circular consumable replacement port 4 is provided at the center of the upper cover 2, and the upper cover 2 is detachably and sealedly connected to a circular top cover 5 at the consumable replacement port 4, wherein the size of the top cover 5 is larger than the size of the consumable replacement port 4. The culture chamber is connected to the inner side of the top cover 5; the stirring chamber is connected to the side of the culture chamber away from the top cover 5, and is suspended in the tank body 1. The top cover 5 is provided with a plurality of sampling ports 6 which are centrally and symmetrically distributed. A sterile sampler is provided at the sampling ports 6 . In this embodiment, the number of the sampling ports 6 is 8 and they are centrally and symmetrically distributed on the top cover 5 .

[0043] The consumables are relatively independent of the tank body 1. The consumables are relatively independent of the tank body 1. The installation of the consumables does not require opening the upper cover 2 of the tank body 1, which will not affect the pipeline of the tank body 1 and is more convenient to install. It is easy to disassemble and replace, reducing the operation steps and time. Therefore, through the reasonable design of the pipeline structure 3 and the consumable replacement port 4, it is convenient for users to maintain the reactor and replace consumables. Since most of the existing stirring devices on the market are polytetrafluoroethylene stirrers, the polytetrafluoroethylene stirrers will wear out after long-term operation, producing black particles or colloids, which affect cell growth; and the stirring device is fixed to the bottom of the tank body 1. Therefore, during use, it needs to be regularly inspected and maintained, and it is not conducive to replacement. The stirring device and the culture chamber are integrated into a disposable product and a disposable stirring structure to reduce foreign matter caused by wear.

[0044] A seal 16 is provided between the consumables replacement port 4 and the top cover 5. An annular mounting groove 22 is provided on the top cover 2 around the consumables replacement port 4. The seal 16 is a rubber sealing ring that has an interference fit within the mounting groove 22. The seal protrudes from the mounting groove 22 and abuts against the side of the top cover 5 closest to the top cover 2. The seal 16 helps improve the sealing performance of the connection between the top cover 5 and the consumables replacement port 4.

[0045] The top cover 5 is provided with a plurality of mounting holes 24 distributed in a circumferential array, and the upper cover 2 is provided with threaded holes 23 at positions corresponding to the mounting holes 24; the top cover 5 and the upper cover 2 are detachably connected and fixed by hand bolts 17 which pass through the mounting holes 24 and are threadedly connected to the threaded holes 23. The hand bolts 17 facilitate manual disassembly and installation, thereby facilitating quick replacement of consumables.

[0046] Reference Figure 3 and Figure 4 The culture chamber includes a cylindrical inner tube 7 made of stainless steel. A fixing hole 25 is defined in the sidewall of the inner tube 7, perpendicular to the central axis of the tank body 1. A fixing member 18 is positioned within the fixing hole 25. The fixing member 18 can be a bolt, which connects the inner tube 7 to the top cover 5. This facilitates the removal, replacement, and installation of the fixed bed 11.

[0047] Reference Figure 5, a bracket is provided in the inner cylinder 7, and a fixed bed 11 is installed on the bracket. The bracket includes a central column 9 and at least two limiting rods 10 fixedly connected to the central column 9. This embodiment is demonstrated by taking two as examples. The fixed bed 11 is centered on the central column 9 and is limited between the limiting rods 10 and the inner cylinder 7; a winding hole 26 is provided on the central column 9 along the axial direction to facilitate the fixation of the side edge of the roll film on the fixed bed 11, so as to facilitate winding on the central column 9 to form a cylindrical fixed bed 11 structure. One end of the central column 9 is fixedly connected to the top cover 5 by a thread, and the other end is fixedly connected to the limiting rod 10. Fixed installation by the central column 9 and the top cover 5 is conducive to improving the stability of the fixed bed 11.

[0048] Multiple first recirculation holes 8 are defined around the sidewall of the inner cylinder 7 near the top cover 5, and the bottom of the inner cylinder 7 communicates with the stirring chamber. The connection between the inner cylinder 7 and the top cover 5 allows for a suspended culture chamber. The bracket supports the fixed bed 11, and the first recirculation holes 8 ensure that the flow of the cell culture fluid is not affected during stirring.

[0049] Reference Figure 6 and Figure 7 The stirring chamber includes a cylindrical chamber body 12, which is integrally formed with the inner tube 7 and is made of stainless steel. The chamber body 12 is located at the end of the inner tube 7 away from the upper cover 2. A magnetic stirring rod 13 is provided in the chamber body 12, and a plurality of circular second reflux holes 14 are arranged in an array at the bottom of the chamber body 12. The chamber body 12 is suspended, integrating the stirring chamber and the culture chamber as a whole. The disposable stirring structure not only helps reduce foreign matter caused by wear and tear, but also facilitates removal and does not easily affect the pipeline layout within the tank body 1.

[0050] In order to increase the effect of magnetic stirring, the bottom of the chamber 12 is provided with a disc-shaped glass cover 15, the second reflux hole 14 is opened on the glass cover 15, and a set gap is reserved between the glass cover 15 and the bottom of the tank body 1, that is, the culture chamber does not contact the bottom of the tank body 1. The glass surface is smooth, and when the magnetic stirring bar 13 rotates on the glass, it can form a relatively uniform vortex, thereby achieving a better stirring effect. Compared with the material of stainless steel, there may be tiny bumps or impurities on the surface of stainless steel, and these factors will affect the stirring effect of the magnetic stirring bar 13 when it rotates on the stainless steel. In addition, the glass surface is smooth and easy to clean, and the residue that may be generated by the magnetic stirring bar 13 during the stirring process can be easily removed.

[0051] The implementation principle of a fixed-bed bioreactor for sterile sampling in an embodiment of the present application is as follows: it can realize the sterile sampling function of a 2-square-meter fixed-bed 11 bioreactor, and no longer relies on a biosafety cabinet. The new structure makes the consumables relatively independent from the tank body 1. The installation of the consumables does not require opening the upper cover 2 of the tank body 1, which will not affect the piping layout of the tank body 1, making installation more convenient. The new structure combines the stirring chamber and the culture chamber into one, and is used as a disposable product, which can effectively solve the adverse effects caused by wear; the new structure can realize the sterile sampling function of the fixed-bed 11 bioreactor; and cell culture can be carried out in an environment other than a biosafety cabinet.

[0052] The present application discloses a sterile sampler:

[0053] Reference Figure 8 and Figure 9 The sterile sampler includes a hose 19. In this embodiment, the hose 19 is made of a transparent material, such as silicone. One end of the hose 19 is sealed and connected to the sampling port 6, and the other end is sealed and connected to the upper plug 28. Inside the hose 19, a lower plug 29 is provided between the sampling port 6 and the upper plug 28; a slit is provided on the lower plug 29, which is not shown in the figure. A sampling strip 21 is provided on the fixed bed 11, and the end of the sampling strip 21 passes through the sampling port 6 and is provided in the hose 19, and the sampling strip 21 is clamped in the slit; a pipe clamp 20 is clamped on the hose 19.

[0054] Before sampling, the structure of the hose 19, such as Figure 8 As shown. When sampling, open the tube clamp 20, remove the upper plug 28, and use tools to pull the sampling strip 21 out of the tank body 1. The transparent hose 19 makes it easy to observe the position of the sampling strip 21. When the sampling strip 21 is pulled to a certain position, the sampling strip 21 of the required length L is located at a section of the lower plug 29 close to the tank body 1; then use a shearing tool to cut the hose 19. In order to ensure sterility, disinfectant will be sprayed at the cut end of the hose 19 after cutting. Then, seal the lower plug 29 at the end of the hose 19 that is still connected to the tank body 1, and use the tube clamp 20 to clamp the hose 19 to ensure the sterility of the tank body 1; the removed sampling strip 21 is wrapped in the hose 19 for subsequent test data analysis. After sampling, the structure of the hose 19 is as shown Figure 9 shown.

[0055] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations on the present application. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present application.

Claims

1. A sterile sampler, characterized in that: The invention comprises a hose (19), one end of which is sealedly connected to a sampling port (6), and the other end of which is sealedly connected to an upper plug (28); a lower plug (29) is provided in the hose (19) between the sampling port (6) and the upper plug (28), and a slit is provided on the lower plug (29); the end of a sampling strip (21) provided on a fixed bed (11) passes through the sampling port (6) and is provided in the hose (19), and the sampling strip (21) is clamped in the slit; and a pipe clamp (20) is clamped on the hose (19).

2. A fixed-bed bioreactor for aseptic sampling, comprising a tank body (1), a stirring chamber and a culture chamber for installing a fixed bed (11), characterized in that: The tank body (1) is provided with an upper cover (2), and a pipeline structure (3) for communicating with the interior of the tank body (1) is provided on the upper cover (2); a consumables replacement port (4) is provided on the upper cover (2), and the upper cover (2) is detachably and sealedly connected to a top cover (5) at the consumables replacement port (4), and the culture chamber is connected to the inner side of the top cover (5); the stirring chamber is arranged in the tank body (1); a sampling port (6) is provided on the top cover (5), and a sterile sampler is provided at the sampling port (6).

3. The fixed-bed bioreactor for sterile sampling according to claim 2, characterized in that: The stirring chamber is connected to a side of the culture chamber away from the top cover (5) and is suspended in the tank body (1).

4. The fixed-bed bioreactor for sterile sampling according to claim 3, characterized in that: The culture chamber comprises an inner cylinder (7), the inner cylinder (7) being connected to the top cover (5), a bracket being provided in the inner cylinder (7), a fixed bed (11) being provided on the bracket, a first reflux hole (8) being provided on a side of the side wall of the inner cylinder (7) close to the top cover (5), and the bottom of the inner cylinder (7) being communicated with the stirring chamber.

5. The fixed-bed bioreactor for sterile sampling according to claim 4, characterized in that: The bracket comprises a central column (9) and at least two limiting rods (10) fixedly connected to the central column (9); the fixed bed (11) is centered on the central column (9) and is limited between the limiting rods (10) and the inner cylinder (7); the central column (9) and the top cover (5) are fixedly connected.

6. The fixed-bed bioreactor for sterile sampling according to claim 4, characterized in that: The stirring chamber comprises a chamber body (12), wherein the chamber body (12) is fixedly connected to an end of the inner cylinder (7) away from the upper cover (2), a stirring rod (13) is arranged in the chamber body (12), and a second reflux hole (14) is arranged at the bottom of the chamber body (12).

7. The fixed-bed bioreactor for sterile sampling according to claim 6, characterized in that: A glass cover (15) is fixedly connected to the bottom of the tank body (12), the second reflux hole (14) is provided on the glass cover (15), and a set gap is reserved between the glass cover (15) and the bottom of the tank body (1).

8. The fixed-bed bioreactor for sterile sampling according to claim 3, characterized in that: A sealing member (16) is sealed between the consumable material replacement port (4) and the top cover (5).

9. The fixed-bed bioreactor for sterile sampling according to claim 3, characterized in that: The top cover (5) and the upper cover (2) are detachably connected and fixed via a plurality of hand-tightened bolts (17).

10. The fixed-bed bioreactor for sterile sampling according to claim 4, characterized in that: The inner cylinder (7) and the top cover (5) are connected via a plurality of fixing members (18).

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