Biological reaction container sampling device
By employing a bioreactor sampling device with components such as a check valve, a breather, and a sealed collection bottle in the bioreactor, the problems of high risk of bacterial contamination, complex operation, and cell lethality risk in existing technologies have been solved, achieving continuous aseptic sampling and improving sampling efficiency and safety.
Patent Information
- Application Number
- CN202422968017.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-03
- Publication Date
- 2025-12-09
- Estimated Expiration
- 2034-12-03
AI Technical Summary
Existing bioreactor sampling processes suffer from high risk of bacterial contamination, complex operation, risk of cell death, and low efficiency.
A sampling device for a bioreactor was designed, which uses components such as a check valve, a breather, and a sealed collection bottle to achieve continuous aseptic sampling. Contamination is avoided by controlling the valve opening and closing sequence, and gravity-fed sampling is used to reduce operation steps and contact with high-temperature steam.
Aseptic sampling was achieved, reducing the risk of contamination, simplifying the operation process, reducing the risk of cell death, and improving sampling efficiency and safety.
Smart Images
Figure CN223646553U_ABST
Abstract
Description
Technical Field
[0001] This utility model belongs to the field of pharmaceutical equipment technology, and in particular relates to a sampling device for a bioreactor. Background Technology
[0002] Animal cell in vitro culture technology drives the development of the biopharmaceutical industry, and is applied in the fields of human vaccines, veterinary vaccines, recombinant proteins, antibodies and fusion proteins, and gene and cell therapy. It is used to culture various cell lines such as CHO cells, HEK 293 cells, Vero cells, BKH cells, and T lymphocytes. The key equipment for expanding cell survival is the bioreactor. In the process of using bioreactor cell culture, it is necessary to take samples for analysis every day or aseptically remove samples for simultaneous small-scale control. Frequent sampling and opening operations can easily increase the risk of bacterial contamination and affect culture failure. The cell culture cycle itself is long and the culture medium is expensive. Bacterial contamination failure causes waste of time and costs. Therefore, ensuring the aseptic sampling process is very important.
[0003] Currently, common stainless steel sampling methods use four-valve vials for connection. The four-valve vials are first centrifuged and sterilized, then reassembled into the bioreactor to sterilize the sampling pipeline. Samples are then taken into sterile vials, and after the sampling valve is closed, the samples are sent for analysis. This process is repeated for the next sampling. While this overall process ensures aseptic sampling, the daily cleaning, offline sterilization, online sterilization, and repeated sampling steps increase labor time. Furthermore, with cells cultured inside the bioreactor and the sampling valve directly connected to the reactor, the daily online SIP steam at high temperatures, conducted through the sampling valve, transfers heat to the cells in the reactor, offsetting the temperature drop. Temperatures exceeding 121°C transfer to localized cells within the reactor tank, potentially causing cell death. The SIP process before daily sampling further increases the frequency and risk of cell death, leading to process failure.
[0004] Currently, common disposable bioreactor bags are equipped with one Luer tube and one thermoplastic tube for sampling. Two sterile syringes are needed for sampling. One syringe is connected to the Luer tube head for aspiration, then the sampling tube clamp is opened to draw about 30ml of sample and discard it. The other syringe is then used to draw the sample and the clamp is closed. The Luer tube head is sterilized with 75% alcohol. Although the above operation is simple, each time the syringe draws a sample, the clamp inside the container needs to be opened. This operation is open, and if the indoor environment is not a clean area, bacteria may contaminate the reverse pipeline and enter the reactor, posing a risk of bacterial contamination under open operation. Utility Model Content
[0005] The technical problem to be solved by this utility model is to overcome the shortcomings of the prior art and provide a sampling device for bioreactors that can achieve continuous aseptic sampling.
[0006] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:
[0007] A sampling device for a bioreactor includes a bioreactor 1 and a sampling pipe 2 connected to the bottom of the bioreactor 1. A third valve 14 is provided between the sampling pipe 2 and the bioreactor 1. A first check valve 3, a first three-way pipe 4, and a second three-way pipe 5 are sequentially connected to the sampling pipe 2. The first three-way pipe 4 is also connected to a second check valve 6. The second check valve 6 is connected to a first respirator 7. The second three-way pipe 5 is connected to a first valve 9 and a second valve 16. The first valve 9 is connected to a collection bottle 11. The collection bottle 11 is sealed and is equipped with a second respirator 12. The second valve 16 is connected to a third check valve 8. The third check valve 8 is connected to a Luer connector 10 to achieve continuous aseptic sampling.
[0008] As a further improvement to the above technical solution:
[0009] Sampling pipe 2 is a flexible hose, and the first valve 9 and the second valve 16 are pipe clamps for easy operation.
[0010] The first respirator 7 and the second respirator 12 are butterfly filters.
[0011] The collection bottle 11 is detachably connected to the sampling pipe 2, and the collection bottle 11 can be replaced.
[0012] The bioreactor 1 is a disposable bioreactor 100, the sampling pipe 2 is a thermoplastic tube, and the bottom of the disposable bioreactor 100 is equipped with a disposable sampling tube 13, which is a thermoplastic tube. The sampling pipe 2 and the disposable sampling tube 13 are connected by a connector for easy docking.
[0013] The third valve 14 is a pipe clamp for easy operation.
[0014] The bioreactor 1 is a stainless steel bioreactor 200. The bottom of the stainless steel bioreactor 200 is equipped with a third valve 14, which is a sampling valve. The third valve 14 is connected to a stainless steel sampling tube 17, which is connected to a sampling pipeline 2.
[0015] The sampling valve is also connected to the sterilization tube 18, which is equipped with a first pneumatic diaphragm valve 19. The stainless steel sampling tube 17 is equipped with a second pneumatic diaphragm valve 20. One end of the second pneumatic diaphragm valve 20 is connected to the sampling pipe 2, and the other end is connected to the drain pipe 21. The drain pipe 21 is equipped with a third pneumatic diaphragm valve 22 and a drain valve 23, which facilitates continuous aseptic sampling after online sterilization.
[0016] A first clamp 24 is provided between the second pneumatic diaphragm valve 20 and the sampling valve, a second clamp 25 is provided between the second pneumatic diaphragm valve 20 and the sampling pipe 2, and a third clamp 15 is provided between the second pneumatic diaphragm valve 20 and the drain pipe 21 for easy connection.
[0017] Both the sterilization tube 18 and the drain tube 21 are stainless steel pipes, which facilitates continuous aseptic sampling after online sterilization.
[0018] Compared with the prior art, the advantages of this utility model are:
[0019] This invention relates to a bioreactor sampling device. The sampling pipe 2 maintains pressure balance with the outside environment. During sampling, the liquid flows into the sampling pipe 2 by gravity. The device has a simple and compact structure, making it easy to operate. By setting a first check valve 3, a second check valve 6, and a third check valve 8, the device prevents the external environment from contaminating the inside of the bioreactor 1 and the sampled liquid. By controlling the opening and closing sequence of the first valve 9, the second valve 16, and the third valve 14, the device ensures that the sample does not come into contact with the inside of the bioreactor, achieving aseptic sampling. A sealed collection bottle 11 is provided to remove residual liquid in a closed environment before sampling. This device can be used to collect residual liquid multiple times, achieving continuous aseptic sampling. Attached Figure Description
[0020] Figure 1 This is a schematic diagram of the structure of a sampling device for a disposable bioreactor according to the present invention;
[0021] Figure 2 This is a schematic diagram of the structure of a stainless steel bioreactor sampling device according to the present invention.
[0022] The reference numerals in the figures of this utility model are as follows: 1. Bioreactor; 100. Disposable bioreactor; 200. Stainless steel bioreactor; 2. Sampling pipe; 3. First check valve; 4. First tee pipe; 5. Second tee pipe; 6. Second check valve; 7. First respirator; 8. Third check valve; 9. First valve; 10. Luer connector; 11. Collection bottle; 12. Second respirator; 13. Disposable sampling tube; 14. Third valve; 15. Third clamp; 16. Second valve; 17. Stainless steel sampling tube; 18. Sterilization tube; 19. First pneumatic diaphragm valve; 20. Second pneumatic diaphragm valve; 21. Drain pipe; 22. Third pneumatic diaphragm valve; 23. Steam trap; 24. First clamp; 25. Second clamp. Detailed Implementation
[0023] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.
[0024] like Figures 1 to 2As shown, a bioreactor sampling device includes a bioreactor 1 and a sampling pipe 2 connected to the bottom of the bioreactor 1. A third valve 14 is provided between the sampling pipe 2 and the bioreactor 1. A first check valve 3, a first three-way pipe 4, and a second three-way pipe 5 are sequentially connected to the sampling pipe 2. The first three-way pipe 4 is also connected to a second check valve 6. The second check valve 6 is connected to a first respirator 7. The second three-way pipe 5 is connected to a first valve 9 and a second valve 16. The first valve 9 is connected to a collection bottle 11. The collection bottle 11 is sealed and is equipped with a second respirator 12. The second valve 16 is connected to a third check valve 8. The third check valve 8 is connected to a Luer connector 10 to achieve continuous aseptic sampling.
[0025] Sampling pipe 2 is connected to the bottom of bioreactor 1. A first respirator 7 and a second respirator 12 are connected to sampling pipe 2 to maintain pressure balance between sampling pipe 2 and the outside. During sampling, the liquid inside bioreactor 1 flows into sampling pipe 2 by gravity. The structure is simple and compact. "Sampling pipe 2 is connected to the bottom of bioreactor 1" means it is connected to the lower part of bioreactor 1, which can be connected to the bottom wall or side wall. Furthermore, bioreactor 1 can be placed on a support or equipped with legs, ensuring that sampling pipe 2 has a certain length in the vertical direction after being connected to bioreactor 1.
[0026] By setting the first check valve 3, it is ensured that the liquid in the sampling pipeline 2 will not flow back into the bioreactor 1, thus preventing the liquid in the bioreactor 1 from being contaminated. By setting the second check valve 6, it is ensured that the liquid will not flow into the first respirator 7, thereby coming into contact with the external environment and causing the sampling liquid to be contaminated. By setting the third check valve 8, it is ensured that the liquid entering the Luer connector will not flow back into the sampling pipeline 2, thus preventing the liquid that came into contact with the external environment from flowing back into the sampling pipeline 2 and causing the sampling pipeline 2 to be contaminated.
[0027] To achieve continuous aseptic sampling, during sampling, first close the second valve 16 and open the first valve 9 and the third valve 14. This allows the liquid in the bioreactor 1 to enter the collection bottle 11, expelling residual liquid and air from the sampling pipe 2. Because the collection bottle 11 is sealed, the liquid in the bioreactor 1 will not come into contact with the external environment. Once the sampling pipe 2 is full of liquid, close the third valve 14 and the first valve 9, then open the second valve 16. Use a syringe with a Luer connector connected to the Luer connector 10 to perform sampling. During sampling, only the liquid in the sampling pipe 2 is drawn, avoiding contact with other liquids in the bioreactor 1 and minimizing the risk of contamination. The above steps are repeated for subsequent sampling. Due to the aseptic nature of the sampling process, repeated sterilization is unnecessary, improving sampling efficiency.
[0028] The sampling pipeline 2 is a flexible tube, and the first valve 9 and the second valve 16 are clamps for easy operation. The entire sampling pipeline 2 can be sterilized offline. The first respirator 7 and the second respirator 12 are butterfly filters to prevent airborne contaminants from contaminating the inside of the sampling pipeline 2. The collection bottle 11 is detachably connected to the sampling pipeline 2 and can be replaced. When large-volume sampling is required, the collection bottle can be placed in a movable laminar flow cart and replaced in a sterile environment. The replaced collection bottle 11 can be used directly for sampling, achieving large-volume sampling. When small-volume sampling is performed, the collection bottle 11 is used for residual liquid collection. Before sampling, the first valve 9 and the third valve 14 are opened to allow the liquid in the bioreactor 1 to enter the collection bottle 11 first, expelling residual liquid and air from the sampling pipeline 2.
[0029] The design enables sterile waste discharge for multiple samples, requiring only one syringe. The extracted samples accurately reflect the condition inside the tank, eliminating sampling problems and malfunctions during the testing process. The overall operation and sampling are convenient, eliminating the need for repeated SIP and reducing the risk of heat transfer from steam through the sampling valve to cells inside the tank, which could cause cell death.
[0030] like Figure 1 As shown, the bioreactor 1 is a disposable bioreactor 100, the sampling pipe 2 is a thermoplastic tube, and the bottom of the disposable bioreactor 100 is provided with a disposable sampling tube 13. The disposable sampling tube 13 is a thermoplastic tube and a third valve 14 is provided on the disposable sampling tube 13. The sampling pipe 2 and the disposable sampling tube 13 are connected by a pipe connector for easy docking. The third valve 14 is a pipe clamp for easy operation.
[0031] like Figure 2 As shown, the bioreactor 1 is a stainless steel bioreactor 200. The bottom of the stainless steel bioreactor 200 is equipped with a third valve 14, which is a sampling valve. The third valve 14 is connected to a stainless steel sampling tube 17, which is connected to a sampling pipeline 2. The sampling valve is also connected to a sterilization tube 18. The sterilization tube 18 is equipped with a first pneumatic diaphragm valve 19, and the stainless steel sampling tube 17 is equipped with a second pneumatic diaphragm valve 20. One end of the second pneumatic diaphragm valve 20 is connected to the sampling pipeline 2, and the other end is connected to a drain pipe 21. The drain pipe 21 is equipped with a third pneumatic diaphragm valve 22 and a drain valve 23 to facilitate continuous aseptic sampling after online sterilization.
[0032] A first clamp 24 is provided between the second pneumatic diaphragm valve 20 and the sampling valve, a second clamp 25 is provided between the second pneumatic diaphragm valve 20 and the sampling pipe 2, and a third clamp 15 is provided between the second pneumatic diaphragm valve 20 and the drain pipe 21 for easy connection.
[0033] Both the sterilization tube 18 and the drain tube 21 are stainless steel pipes, which facilitates continuous aseptic sampling after online sterilization.
[0034] Sampling pipe 2 is sterilized offline, while sampling valve and stainless steel bioreactor 200 are sterilized online. During sterilization, the second pneumatic diaphragm valve 20 is closed, and the first pneumatic diaphragm valve 19 and the third pneumatic diaphragm valve 22 are opened. Steam enters the sampling valve through sterilization pipe 18 and then enters stainless steel sampling pipe 17. Condensate is discharged through drain valve 23 on drain pipe 21, thus achieving sterilization. Then, the offline sterilized sampling pipe 2 is connected to stainless steel sampling pipe 17 through second clamp 25, and aseptic sampling can be performed.
[0035] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make many possible variations and modifications to the present invention, or modify it into equivalent embodiments, without departing from the scope of the present invention. Therefore, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the present invention, should fall within the protection scope of the present invention.
Claims
1. A sampling device for a bioreactor, characterized in that, The device includes a bioreactor (1) and a sampling pipe (2) connected to the bottom of the bioreactor (1). A third valve (14) is provided between the sampling pipe (2) and the bioreactor (1). A first check valve (3), a first three-way pipe (4), and a second three-way pipe (5) are connected sequentially to the sampling pipe (2). The first three-way pipe (4) is also connected to a second check valve (6). The second check valve (6) is connected to a first respirator (7). The second three-way pipe (5) is connected to a first valve (9) and a second valve (16). The first valve (9) is connected to a collection bottle (11). The collection bottle (11) is sealed and a second respirator (12) is provided on the collection bottle (11). The second valve (16) is connected to a third check valve (8). The third check valve (8) is connected to a Luer connector (10).
2. The sampling device for a bioreactor as described in claim 1, characterized in that, The sampling pipe (2) is a flexible hose, and the first valve (9) and the second valve (16) are pipe clamps.
3. The sampling device for a bioreactor as described in claim 1, characterized in that, The first respirator (7) and the second respirator (12) are butterfly filters.
4. The sampling device for a bioreactor as described in claim 1, characterized in that, The collection bottle (11) is detachably connected to the sampling pipe (2).
5. A sampling device for a bioreactor as described in claim 1, characterized in that, The bioreactor (1) is a disposable bioreactor (100), the sampling pipe (2) is a thermoplastic tube, and a disposable sampling tube (13) is provided at the bottom of the disposable bioreactor (100). The disposable sampling tube (13) is a thermoplastic tube, and the sampling pipe (2) and the disposable sampling tube (13) are connected by a pipe connector.
6. A bioreactor sampling device as described in claim 5, characterized in that, The third valve (14) is a pipe clamp.
7. A sampling device for a bioreactor as described in claim 1, characterized in that, The bioreactor (1) is a stainless steel bioreactor (200). The bottom of the stainless steel bioreactor (200) is provided with a third valve (14). The third valve (14) is a sampling valve. The third valve (14) is connected to a stainless steel sampling tube (17). The stainless steel sampling tube (17) is connected to a sampling pipeline (2).
8. A sampling device for a bioreactor as described in claim 7, characterized in that, The third valve (14) is also connected to the sterilization tube (18), which is equipped with a first pneumatic diaphragm valve (19) and a second pneumatic diaphragm valve (20) on the stainless steel sampling tube (17). One end of the second pneumatic diaphragm valve (20) is connected to the sampling pipe (2) and the other end is connected to the drain pipe (21). The drain pipe (21) is equipped with a third pneumatic diaphragm valve (22) and a drain valve (23).
9. A bioreactor sampling device as described in claim 8, characterized in that, A first clamp (24) is provided between the second pneumatic diaphragm valve (20) and the sampling valve, a second clamp (25) is provided between the second pneumatic diaphragm valve (20) and the sampling pipe (2), and a third clamp (15) is provided between the second pneumatic diaphragm valve (20) and the drain pipe (21).
10. A sampling device for a bioreactor as described in claim 8, characterized in that, Both the sterilization pipe (18) and the drainage pipe (21) are stainless steel pipes.