Advanced online sterile sampling system

Abstract

The utility model discloses an advanced on-line sterile sampling system, which belongs to the design field of on-line sterile sampling systems in biological product production process, and comprises a bioreactor which is sequentially connected to a reactor sampling pipeline pipe clamp, a first one-way valve, a second one-way valve and a third one-way valve, the rear portion of the third one-way valve is connected to a Luer taper, and then the Luer taper is connected to an injector. A sterile sampling bag pipe clamp is connected between the first one-way valve and the second one-way valve; a waste liquid bag pipe clamp is connected between the second one-way valve and the third one-way valve; the sterile sampling bag pipe clamp is connected to a sterile sampling bag, and the waste liquid bag pipe clamp is connected to a waste liquid bag. According to the utility model, the bioreactor cannot be polluted in the sampling process through simple operation under the working condition of high-frequency sampling, the sampling of sterile samples is supported, and the actual production requirements are met.

Description

Technical Field

[0001] The utility model belongs to the design field of an on-line aseptic sampling system in the production process of biological products. Specifically, it relates to a sampling system using a special sampling system in glass bioreactors, disposable bioreactors, stainless steel bioreactors, etc. Background Technique

[0002] The production of biological products uses mammalian cells as hosts, and a culture medium is used in a bioreactor for cell passage amplification and final reactor culture to express the target product. Usually, from cell resuscitation to the final harvest of the target protein, the entire process takes nearly 40 days. Since the culture medium is rich in nutrients and is extremely vulnerable to microbial contamination, the entire cell culture process needs to ensure absolute sterility to ensure the normal growth, metabolism, and expression of the target protein of the cells.

[0003] According to the requirements of production process monitoring, parameter feedback regulation, and regulatory regulations, two types of sample samplings are usually required during the production process. The first type requires taking out the culture solution from the bioreactor regularly (such as once a day) to detect process parameters such as cell density, cell viability, and biochemical parameters, and perform process regulation according to the relevant process parameter levels. Such samples need to be ensured to be uniform and without residues from the previous sampling to ensure that the taken samples can truly represent the real-time state of the liquid in the bioreactor. The second type is that in the later stage of cell culture, a large volume of aseptic culture solution needs to be taken from the bioreactor according to relevant regulatory requirements to detect key indicators such as sterility, mycoplasma, and virus contamination. Such samples need to be ensured to be strictly aseptic to avoid false positive abnormalities in sample detection.

[0004] The disposable bioreactor uses a disposable plastic bag, and the sampling system uses a standard configuration of NA interface and TC connector. After the whole set of reactors is assembled, it is irradiated with gamma rays to ensure that the whole system is in an aseptic state. When sampling the daily process parameter detection samples, a disposable sterile syringe with a matching thread is docked with the sampling port, and the culture solution is drawn into the syringe for sampling. Since the syringe thread and the bioreactor sampling system interface are carried out in an ordinary environment, absolute sterility cannot be guaranteed, and the liquid in the syringe can be pushed back into the bioreactor, so there is a greater risk of contamination. When sampling aseptic samples, aseptic welding is performed with a sampling bottle through a heat shrinkable tube. After the sample is aseptically taken out, the heat shrinkable tube is aseptically sealed with a tube sealer. However, each sampling requires repeating the aseptic connection and sealing operations, and the aseptic risk during the connection process accumulates continuously.

[0005] The stainless-steel bioreactor adopts a valve group sampling system. Before each sampling, the sampling system of the reactor is subjected to SIP using high-pressure steam. After cooling, the sampling bottle is docked with the reactor sampling system, and then the interface is subjected to SIP. After cooling, sampling is carried out. After sampling is completed, the sampling bottle is disconnected, and then the reactor sampling system is subjected to SIP. The sampling process is extremely complex, time-consuming, and the cooling water generated during the SIP process is likely to enter the culture medium, resulting in dilution of the culture medium.

[0006] The glass bioreactor is relatively small in scale, with a volume usually less than 10 L. Generally, the sampling bottle is connected during the assembly of the reactor, and then they are transferred to the high-pressure sterilization cabinet for off-line sterilization together. During sampling, the culture medium is pumped into the sampling bottle through a syringe, and then the sampling bottle is replaced to complete sampling. The process of replacing the sampling bottle is an open operation, with a relatively high risk of contamination, and it is impossible to take sterile samples. Utility Model Content

[0007] To solve the problems existing in the existing sampling technologies of the above three bioreactors, the utility model applicant of this application has conducted a large amount of research and developed an on-line aseptic sampling system for bioreactors. The sampling system can support simple operation, and under the condition of high-frequency sampling, the sampling process will not pose a contamination risk to the bioreactor, and it supports the sampling of sterile samples, meeting the requirements of actual production.

[0008] The present utility model is implemented as follows:

[0009] An advanced on-line aseptic sampling system, the sampling system includes: a bioreactor, which is sequentially connected to a clamp of the reactor sampling pipeline, a first one-way valve, a second one-way valve, and a third one-way valve behind the bioreactor. The third one-way valve is connected to a Luer connector, and the Luer connector is further connected to a syringe; an aseptic sampling bag clamp is connected between the first one-way valve and the second one-way valve; a waste liquid bag clamp is connected between the second one-way valve and the third one-way valve; the aseptic sampling bag clamp is further connected to an aseptic sampling bag, and the waste liquid bag clamp is further connected to a waste liquid bag.

[0010] Further, the bioreactor is connected to the clamp of the reactor sampling pipeline through a thermoplastic tube; the aseptic sampling bag clamp is connected to the waste liquid bag clamp through a thermoplastic tube; the aseptic sampling bag is connected to the waste liquid bag through a thermoplastic tube; other connections are all made of silicone tubes; the thermoplastic tube can be aseptically welded or heat-sealed.

[0011] Further, the entire system is aseptically processed by gamma irradiation.

[0012] Further, the sampling of the system includes non-sterile sample sampling and sterile sample sampling: The method for non-sterile sample sampling is:

[0013] 1), aseptically connect the thermoplastic tube of the sampling system to the thermoplastic tube on the sampling pipeline of the bioreactor; the thermoplastic tube can be aseptically welded or heat-sealed.

[0014] 2), dock the syringe with the sampling interface of the Luer connector. After opening the clamp of the waste liquid bag and the clamp of the reactor sampling pipeline, draw the liquid material into the waste liquid bag through the syringe to clean the residue of the previous sampling in the pipeline. If this operation is not required for the first sampling, close the clamp of the waste liquid bag after completion.

[0015] 3), draw the liquid material into the syringe through the syringe, close the clamp of the reactor sampling pipeline, disinfect the threaded interface with 75% ethanol, and then cover the lid.

[0016] In aseptic sample sampling, the sampling method includes the following steps:

[0017] 1), aseptically connect the thermoplastic tube of the sampling system to the thermoplastic tube on the sampling pipeline of the bioreactor. The thermoplastic tube is the thermoplastic tube between the bioreactor and the clamp of the reactor sampling pipeline, the thermoplastic tube between the clamp of the aseptic sampling bag and the clamp of the waste liquid bag, and the thermoplastic tube between the aseptic sampling bag and the waste liquid bag.

[0018] 2), dock the syringe with the Luer connector. After opening the clamp of the waste liquid bag and the clamp of the reactor sampling pipeline, draw the liquid material into the waste liquid bag through the syringe to clean the residue of the previous sampling in the pipeline (this operation is not required for the first sampling), and close the clamp of the waste liquid bag after completion.

[0019] 3), open the clamp of the aseptic sampling bag, draw the liquid material into the aseptic sampling bag through the syringe, close the clamp of the reactor sampling pipeline, close the clamp of the aseptic sampling bag, use an aseptic tube sealer to seal the tube, and complete the sampling after disconnection.

[0020] 4) Disinfect the threaded interface with 75% ethanol and then cover the lid.

[0021] Compared with the traditional sampling method, the beneficial effects of the present utility model are as follows:

[0022] 1), only one aseptic welding is required between the system of the present utility model and the reactor, and aseptic sampling can be quickly achieved through simple operation of the syringe during each sampling.

[0023] 2), since a one-way valve is configured between the reactor sampling pipeline, the aseptic sampling bag, the waste liquid bag, and the Luer interface, the culture solution always flows outward during the sampling process, effectively solving the problem of backflow of the culture solution, and ensuring the aseptic guarantee level under the working conditions of long-term and high-frequency sampling.

[0024] 3), A sterile sampling bag is configured between the first one-way valve and the second one-way valve, which can ensure that the obtained sample is always in a sterile state. A certain number of sterile sampling bags can be configured on this pipeline according to the usage requirements. After each sampling, disconnect the pipeline and seal it for storage.

[0025] 4), A waste liquid bag is configured between the second one-way valve and the third one-way valve. Before each sampling, first pump about 2 pipeline volumes of the culture medium into the waste liquid bag, which can wash the residual liquid in the pipeline from the previous time. Description of the Drawings

[0026] Figure 1 It is a structural diagram of an advanced on-line sterile sampling system of the present utility model;

[0027] Among them, 1 - bioreactor, 2 - reactor sampling pipeline clamp, 3 - first one-way valve, 4 - second one-way valve, 5 - third one-way valve, 6 - syringe, 7 - sterile sampling bag clamp, 8 - waste liquid bag clamp, 9 - sterile sampling bag, 10 - waste liquid bag, 11 - Luer connector. Detailed Embodiments

[0028] To make the purpose, technical solutions and effects of the present utility model clearer and more definite, the following examples are listed to further elaborate on the present utility model in detail. It should be noted that the specific embodiments described herein are only used to explain the present utility model and are not used to limit the present utility model.

[0029] As Figure 1 shown, the sampling system of the present utility model includes: a bioreactor 1, which is sequentially connected to a reactor sampling pipeline clamp 2, a first one-way valve 3, a second one-way valve 4, and a third one-way valve 5 behind the bioreactor 1. A Luer connector 11 is connected behind the third one-way valve 5, and the Luer connector 11 is further connected to a syringe 6; a sterile sampling bag clamp 7 is connected between the first one-way valve 3 and the second one-way valve 4; a waste liquid bag clamp 8 is connected between the second one-way valve 4 and the third one-way valve 5; the sterile sampling bag clamp 7 is further connected to a sterile sampling bag 9, and the waste liquid bag clamp 8 is further connected to a waste liquid bag 10.

[0030] The bioreactor 1 and the reactor sampling pipeline clamp 2 are connected by a thermoplastic tube; the sterile sampling bag clamp 7 and the waste liquid bag clamp 8 are connected by a thermoplastic tube; the sterile sampling bag 9 and the waste liquid bag 10 are connected by a thermoplastic tube; other connections are all made of silicone tubes; the thermoplastic tube can be aseptically welded or heat-sealed. The entire system is aseptically treated by gamma irradiation.

[0031] The sampling of the system of the present utility model includes non-sterile sample sampling and sterile sample sampling:

[0032] The described non-sterile sample sampling method is as follows:

[0033] 1), aseptically connect the thermoplastic tube of the sampling system to the thermoplastic tube on the sampling pipeline of the bioreactor 1; the thermoplastic tube can be aseptically welded or heat-sealed;

[0034] 2), dock the syringe 6 with the sampling interface of the Luer connector 11, open the waste liquid bag clamp 8 of the waste liquid bag 10 and the reactor sampling pipeline reactor sampling pipeline clamp 2 of the bioreactor 1, then draw the liquid material into the waste liquid bag 10 through the syringe 6 to clean the residue of the previous sampling in the pipeline. If it is the first sampling, this operation is not required. After completion, close the waste liquid bag clamp 8 of the waste liquid bag 10;

[0035] 3), draw the liquid material into the syringe through the syringe 6, close the reactor sampling pipeline clamp 2, disinfect the threaded interface with 75% ethanol and then cover it with a lid;

[0036] In the aseptic sample sampling, the described sampling method includes the following steps:

[0037] 1), aseptically connect the thermoplastic tube of the sampling system to the thermoplastic tube on the sampling pipeline of the bioreactor. The thermoplastic tubes are the thermoplastic tube between the bioreactor 1 and the reactor sampling pipeline clamp 2, the thermoplastic tube between the aseptic sampling bag clamp 7 and the waste liquid bag clamp 8, and the thermoplastic tube between the aseptic sampling bag 9 and the waste liquid bag 10;

[0038] 2), dock the syringe 6 with the Luer connector 11, open the waste liquid bag clamp 8 and the reactor sampling pipeline clamp 2, then draw the liquid material into the waste liquid bag through the syringe to clean the residue of the previous sampling in the pipeline (this operation is not required for the first sampling). After completion, close the waste liquid bag clamp 8;

[0039] 3), open the aseptic sampling bag clamp 7, draw the liquid material into the aseptic sampling bag through the syringe, close the reactor sampling pipeline clamp 2, close the aseptic sampling bag clamp 7, and use an aseptic tube sealer to seal the tube. After disconnection, the sampling is completed;

[0040] 4), disinfect the threaded interface with 75% ethanol and then cover it with a lid.

[0041] The above is only the preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements can still be made, and these improvements should also be regarded as the protection scope of the present invention.

Claims

1. An advanced online aseptic sampling system, characterized in that: The sampling system comprises: a bioreactor (1), the bioreactor (1) being connected in sequence to a reactor sampling pipeline clamp (2), a first one-way valve (3), a second one-way valve (4), and a third one-way valve (5), the third one-way valve (5) being connected to a Luer connector (11), and the Luer connector (11) being connected to a syringe (6); A sterile sampling bag tube clamp (7) is connected between the first one-way valve (3) and the second one-way valve (4); a waste liquid bag tube clamp (8) is connected between the second one-way valve (4) and the third one-way valve (5); the sterile sampling bag tube clamp (7) is further connected to the sterile sampling bag (9), and the waste liquid bag tube clamp (8) is further connected to the waste liquid bag (10).

2. The advanced online aseptic sampling system according to claim 1, characterized in that: The The bioreactor (1) and the reactor sampling pipeline clamp (2) are connected via a thermoplastic tube; the sterile sampling bag clamp (7) and the waste liquid bag clamp (8) are connected via a thermoplastic tube; the sterile sampling bag (9) and the waste liquid bag (10) are connected via a thermoplastic tube; the other connections are all silicone tube connections; the thermoplastic tube can be aseptically welded or melt-sealed.

3. An advanced online aseptic sampling system according to any one of claims 1 to 2, characterized in that: The entire system is sterilized by gamma irradiation.

4. An advanced online aseptic sampling system according to any one of claims 1 to 2, characterized in that: The sampling of the system includes non-sterile sample sampling and sterile sample sampling.

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