Online sterile sampling device for fermentation process

Abstract

The utility model relates to the technical field of fermentation sterile sampling, and discloses an online sterile sampling device for a fermentation process, which is provided with a bioreactor, a sampling component, a liquid pumping component, a back flushing component, a quantifying component, a sample receiving group and a control device, the liquid pumping assembly is provided with a pinch valve which is connected to the other end of the sampling assembly and is used for acquiring to-be-sampled liquid, and a first three-way joint and a first peristaltic pump which are sequentially connected to the pinch valve; the reverse blowing assembly is provided with a first gas storage device with a first electromagnetic valve and a second gas storage device with a second electromagnetic valve; the quantifying assembly comprises quantifying pipes which are sequentially connected to the first peristaltic pump; one interface of a third three-way joint of the sample receiving assembly is sequentially connected with a one-way valve, a second peristaltic pump and a cleaning liquid bottle, and the other interface of the third three-way joint is connected with a sampling pool. According to the utility model, full-closed online sterile sampling automatic operation with a short sampling period is realized, the sampling efficiency is effectively improved, sampling consumables are saved, and the like.

Description

Technical Field

[0001] The utility model relates to the technical field of fermentation aseptic sampling, in particular to an online aseptic sampling device used in a fermentation process. Background Art

[0002] The statements in this section are merely to provide background technology related to the present utility model disclosure and do not necessarily constitute prior art.

[0003] In the existing fermentation process and cell culture industry, manual sampling is generally adopted, that is, the culture fluid is extracted from the sampling port of the bioreactor with a syringe, stored in a centrifuge tube, and then the culture fluid is centrifuged with a centrifuge, and the supernatant is filtered with a 0.2μm filter membrane, and finally the processed sample is manually placed in the detection instrument for detection.

[0004] The above prior art has the following defects:

[0005] First, sampling and sample post-processing operations are cumbersome and time-consuming, and cannot meet the requirements of high-frequency sampling;

[0006] Second, manual sampling may pose a risk of contamination, especially when the operation is improper or the operator is unskilled, which may cause external microorganisms to enter the reaction system and affect the fermentation quality;

[0007] Third, the sampling operation process is fixed and requires 24-hour on-duty personnel, and the entire manual operation results in a large waste of human resources;

[0008] Fourth, manual sampling has time uncertainty and cannot fully meet the time parallelism requirements of fermentation sampling.

[0009] In order to solve the above technical problems, the technical solution is hereby proposed as follows. Utility Model Content

[0010] In order to overcome the shortcomings of the above-mentioned prior art, the utility model provides an online aseptic sampling device for fermentation process that realizes automated operation, makes the sampling operation intelligent, realizes full closure, shortens the sampling cycle, effectively improves the sampling efficiency, and reduces human observation, measurement and resource consumption.

[0011] The technical solution adopted by the utility model is: an online aseptic sampling device for a fermentation process, which has a bioreactor, and is characterized in that the online aseptic sampling system also has at least:

[0012] A sampling assembly, one end of which extends below the liquid surface of the bioreactor; and

[0013] A liquid extraction assembly, the liquid extraction assembly comprising a pinch valve connected to the other end of the sampling assembly to obtain the liquid to be sampled, and a first three-way connector and a first peristaltic pump connected in sequence to the pinch valve; and

[0014] A backflush assembly, the backflush assembly comprising a first gas storage device with a first solenoid valve and a second gas storage device with a second solenoid valve, the first solenoid valve and the second solenoid valve are respectively connected to the first three-way connector through a second three-way interface;

[0015] A quantitative component, the quantitative component comprising a quantitative tube sequentially connected to the first peristaltic pump, the quantitative tube having a liquid path bubble sensor; and

[0016] A sample receiving component, wherein the sample receiving component has a third three-way joint configured to be connected to the quantitative tube, one interface of the third three-way joint is connected to the one-way valve, the second peristaltic pump and the cleaning liquid bottle in sequence to obtain the cleaning liquid in the cleaning liquid bottle, and the other interface of the third three-way joint is connected to the sampling pool; and

[0017] A control device is provided, wherein the control device is respectively connected to the clamp valve, the first peristaltic pump, the first solenoid valve, the second solenoid valve, the liquid path bubble sensor and the second peristaltic pump to control the opening and closing states thereof.

[0018] In the present technical solution, three sockets, namely a liquid inlet, a liquid outlet and an overflow port, are arranged on the sampling pool, wherein the overflow port and the liquid outlet are respectively connected to the waste liquid interface through the third peristaltic pump and the fourth peristaltic pump to discharge the waste liquid, thereby automatically realizing the discharge of the cleaning liquid in the sampling pool three.

[0019] In the technical solution, the length of the quantitative tube of the quantitative component is changed according to the size of the sampling volume, so as to control the maximum sampling volume of the quantitative component.

[0020] In the technical solution, the pinch valve is an electromagnetic pulse pinch valve, and its working state is normally closed (NC). The use of the normally closed (NC) electromagnetic pulse pinch valve can ensure the sealing of the liquid pipeline.

[0021] In the present technical solution, the first gas storage device has a first gas container, and a first gas filter and a first gas outlet respectively connected to the first gas container; the second gas storage device has a second gas container, and a second gas filter and a second gas outlet respectively connected to the second gas container; wherein: the first gas filter and the second gas filter are respectively directly connected to the atmosphere or are the inlets of the second gas storage device.

[0022] In the present technical solution, the bioreactor, sampling assembly, liquid extraction assembly, backflush assembly, quantitative assembly and sample receiving assembly are connected by pipelines, and the connecting pipelines are all silicone pipelines or PU gas pipelines.

[0023] In the present technical solution, the bioreactor is a culture tank made of glass or stainless steel.

[0024] Compared with the prior art, the beneficial effects of the utility model are:

[0025] 1. The control device is used to respectively connect the clamp valve, the first peristaltic pump, the first solenoid valve, the second solenoid valve, the liquid path bubble sensor and the second peristaltic pump to control their opening and closing states, completing all steps such as the rinsing step, the sampling step, and the back-blowing step of aseptic sampling in the fermentation process, thereby realizing the full-automatic control of online aseptic sampling in the fermentation process and the full-process automated sampling.

[0026] 2. The liquid extraction component and the back-blowing component are connected to the bioreactor and the sampling pool respectively, and the back-blowing component provides gas power to the sampling and back-blowing processes of the two through the first gas storage device and the second gas storage device respectively, thereby realizing full process control.

[0027] 3. A liquid path bubble sensor is arranged in the quantitative tube of the quantitative component, so that the liquid parameters in the quantitative tube can be fed back to the control device in time, which is convenient for observing and controlling the sampling process, and the sampling accuracy and efficiency are high.

[0028] The utility model discloses an online aseptic sampling device for a fermentation process, which realizes the automated operation of online aseptic sampling, makes the sampling operation intelligent, realizes full closure, has a short sampling cycle, and can be completed in a periodic manner, thereby effectively improving the sampling efficiency, reducing human observation and measurement, and saving sampling consumables. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 A schematic diagram of the structure of an embodiment of an online aseptic sampling device for a fermentation process;

[0030] Figure 2 A schematic diagram of a structure diagram of an embodiment of a backflush assembly;

[0031] Figure 3 is a structural diagram of an embodiment of a sampling assembly;

[0032] Among them: 1-bioreactor, 2-sampling assembly;

[0033] 3-liquid extraction assembly, 301-pinch valve, 302-first peristaltic pump, 303-first three-way connector;

[0034] 4-backflush assembly, 401-high pressure gas interface, 402-first solenoid valve, 403-second solenoid valve, 404-first gas storage device, 405-second gas storage device, 406-second three-way interface, 404-1-first gas container, 404-2-first gas filter, 404-3-first gas outlet, 405-1-second gas container, 405-2-second gas filter, 405-3-second gas outlet;

[0035] 5-quantitative component, 501-liquid path bubble sensor, 502-quantitative tube;

[0036] 6-sample receiving assembly, 601-waste liquid interface, 602-cleaning liquid bottle, 603-sampling pool, 604-second peristaltic pump, 605-third peristaltic pump, 606-fourth peristaltic pump, 607-third three-way connector, 608-fourth three-way connector, 609-one-way valve;

[0037] 7- Control device. DETAILED DESCRIPTION

[0038] The embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0039] In the description of the present invention, it should be understood that the terms "center", "up", "down", "front", "back", "left", "right", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the combination or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. In addition, in the description of the embodiments of the present invention, the device positional relationships such as "up", "down", "front", "back", "left", "right", etc. in all the drawings are based on the orientation or positional relationship shown in the drawings. Figure 1 As standard.

[0040] like Figure 1 As shown, an online aseptic sampling device for a fermentation process has a bioreactor 1, characterized in that the online aseptic sampling system also has at least:

[0041] A sampling assembly 2, one end of which extends below the liquid surface of the bioreactor 1; and

[0042] A liquid extraction component 3, wherein the liquid extraction component 3 comprises a pinch valve 301 connected to the other end of the sampling component 2 to obtain the liquid to be sampled, and a first three-way connector 303 and a first peristaltic pump 302 which are sequentially connected to the pinch valve 301; and

[0043] A backflush assembly 4, wherein the backflush assembly 4 comprises a first gas storage device 404 with a first solenoid valve 402 and a second gas storage device 405 with a second solenoid valve 403, wherein the first solenoid valve 402 and the second solenoid valve 403 are respectively connected to the first three-way connector 303 through a second three-way interface 406;

[0044] A quantitative component 5, the quantitative component 5 comprises a quantitative tube 502 which is sequentially connected to the first peristaltic pump 302, the quantitative tube 502 having a liquid path bubble sensor 501; and

[0045] A sample receiving component 6, wherein the sample receiving component 6 has a third three-way connector 607 configured to be connected to the quantitative tube 502, wherein one interface of the third three-way connector 607 is connected to a one-way valve 609, a second peristaltic pump 604 and a cleaning liquid bottle 602 in sequence to obtain the cleaning liquid in the cleaning liquid bottle 602, and another interface of the third three-way connector 607 is connected to a sampling pool 603; and

[0046] The control device 7 is connected to the clamp valve 301, the first peristaltic pump 302, the first solenoid valve 402, the second solenoid valve 403, the liquid path bubble sensor 501 and the second peristaltic pump 604 to control their opening and closing states. The first peristaltic pump 302, the second peristaltic pump 604, the third peristaltic pump 605 and the fourth peristaltic pump 606 of the utility model are used to ensure accurate control of the amount of liquid pumped or discharged for the components connected thereto, with high stability, which is conducive to improving the sampling efficiency.

[0047] In the specific implementation process, Figure 3 As shown, the sampling component 2 can use the device described in "A sterile online sampling device for fermentation process Application No.: 2023230309979".

[0048] In some embodiments, the sampling pool 603 is provided with three ports: a liquid inlet, a liquid outlet and an overflow port, wherein the overflow port and the liquid outlet are respectively connected to the waste liquid interface 601 through a third peristaltic pump 605 and a fourth peristaltic pump 606 to discharge waste liquid, thereby automatically discharging the cleaning liquid in the sampling pool 603. In the specific implementation process, the sample receiving component 6 also includes a waste liquid interface 601, a cleaning liquid bottle 602, a sampling pool 603, a second peristaltic pump 604, a third peristaltic pump 605, a fourth peristaltic pump 606, a third three-way joint 607, a fourth three-way joint 608, and a one-way valve 609; the outlet of the waste liquid interface 601 is connected to the waste liquid outlet, and the inlet of the waste liquid interface 601 is connected to the c port of the fourth three-way joint 608; the cleaning liquid bottle 602 is connected to the inlet of the second peristaltic pump 604 through a pipeline; the liquid inlet of the sampling pool 603 is connected to the b port of the third three-way joint 607, the liquid outlet of the sampling pool 603 is connected to the inlet of the third peristaltic pump 605, and the overflow port of the sampling pool 603 is connected to the inlet of the fourth peristaltic pump 606; the outlet of the second peristaltic pump 604 is connected to the one-way The inlet of the second peristaltic pump 604 is connected to the cleaning liquid bottle 602; the inlet of the third peristaltic pump 605 is connected to the liquid outlet of the sampling pool 603, and the outlet of the third peristaltic pump 605 is connected to the port a of the fourth three-way joint 608; the inlet of the fourth peristaltic pump 606 is connected to the overflow port of the sampling pool 603, and the outlet of the fourth peristaltic pump 606 is connected to the port b of the fourth three-way joint 608; the port a of the third three-way joint 607 is the first end of the sample receiving component 6, the port b of the third three-way joint 607 is connected to the liquid inlet of the sampling pool 603, and the port c of the third three-way joint 607 is connected to the outlet of the one-way valve 609; the second peristaltic pump 604, the third peristaltic pump 605, and the fourth peristaltic pump 606 are all connected to the control device 7. The port a and port b of the third three-way joint 607 and the port a and port b of the fourth three-way joint 608 mentioned here all refer to one of the interfaces of the three-way joint.

[0049] In some embodiments, the length of the quantitative tube 502 of the quantitative component 5 is changed according to the sampling volume so as to control the maximum sampling volume of the quantitative component 5. In the specific implementation process, the liquid path bubble sensor 501 is buckled on the wall of the quantitative tube 502 by a slot.

[0050] In some embodiments, the pinch valve 301 is an electromagnetic pulse pinch valve, and its working state is normally closed (NC). Using the normally closed (NC) electromagnetic pulse pinch valve 301 can ensure the sealing of the liquid pipeline.

[0051] In some embodiments, see Figure 1 as well as Figure 2The first gas storage device 404 has a first gas container 404-1, and a first gas filter 404-2 and a first gas outlet 404-3 respectively connected to the first gas container 404-1; the second gas storage device 405 has a second gas container 405-1, and a second gas filter 405-2 and a second gas outlet 405-3 respectively connected to the second gas container 405-1; wherein: the first gas filter 404-2 and the second gas filter 405-2 are respectively directly connected to the atmosphere or are the entrances of the second gas storage device 405. During the specific implementation process, the inlet of the high-pressure gas interface 401 is connected to high-pressure air, and the outlet of the high-pressure gas interface 401 is connected to the inlet of the second gas storage device 405; the gas inlet of the first solenoid valve 402 is connected to the outlet of the first gas storage device 404, and the gas outlet of the first solenoid valve 402 is connected to the port a of the second three-way interface 406; the gas inlet of the second solenoid valve 403 is connected to the outlet of the second gas storage device 404, and the gas outlet of the second solenoid valve 403 is connected to the port b of the second three-way interface 406; the port c of the second three-way interface 406 is the third end of the backblowing assembly 4.

[0052] In some embodiments, the bioreactor 1, sampling component 2, liquid extraction component 3, backflush component 4, quantitative component 5 and sample receiving component 6 are connected by pipelines, and the connecting pipelines are all silicone pipelines or PU gas pipelines.

[0053] In some embodiments, the bioreactor 1 is a culture tank made of glass or stainless steel.

[0054] When using the above embodiment, follow the steps below:

[0055] 1. Rinse step: When in use, start the first peristaltic pump 302, start the fourth peristaltic pump 606, and open the clamping valve 301 after the set time is up. The extracted liquid flows to the liquid path bubble sensor 501 through the sampling component 2, the clamping valve 301, the first three-way connector 303, the first peristaltic pump 302, and the quantitative tube 502. When the liquid path bubble sensor 501 detects the liquid flowing through, open the first solenoid valve 402, close the clamping valve 301, and send the extracted liquid in the silicone tube between the first three-way connector 303 and the liquid path bubble sensor 501 into the sampling pool 603; close the first peristaltic pump 302, close the first solenoid valve 402, and start the third peristaltic pump 605 to discharge the extracted liquid in the sampling pool 603 from the waste liquid interface 601 to complete the rinsing step.

[0056] 2. Sampling step: turn off the third peristaltic pump 605, turn on the first peristaltic pump 302, open the pinch valve 301 after the set time is up, and the extracted liquid flows to the liquid path bubble sensor 501 through the sampling component 2, the pinch valve 301, the first three-way connector 303, the first peristaltic pump 302, and the quantitative tube 502. When the liquid path bubble sensor 501 detects that the extracted liquid flows through, open the first solenoid valve 402, close the pinch valve 301, and send the extracted liquid in the silicone tube between the first three-way connector 303 and the liquid path bubble sensor 501 into the sampling pool 603. Turn off the first peristaltic pump 302 and the first solenoid valve 402. This is the sampling step.

[0057] 3. Blowback step: open the pinch valve 301, open the second solenoid valve 403, introduce 0.2 MPa high-pressure gas, and blow the silicone pipeline from the first three-way connector 303 to the sampling component 2 and the liquid taken out from the sampling component 2 back into the bioreactor 1. After the set time is up, close the second solenoid valve 403, close the pinch valve 301, and complete the blowback step.

[0058] 4. Cleaning step: After the extraction liquid in the sampling pool 603 is taken out, start the third peristaltic pump 605 to discharge the residual extraction liquid in the sampling pool 603 from the waste liquid interface 601, turn off the third peristaltic pump 605, start the second peristaltic pump 604, suck the cleaning liquid in the cleaning liquid bottle 602 into the sample pool, turn off the second peristaltic pump 604, start the third peristaltic pump 605, and discharge the cleaning liquid in the sampling pool 603 from the waste liquid interface 601. This is the cleaning step. Repeat the above cleaning steps three times to complete the cleaning operation.

[0059] 5. Online aseptic sampling steps: Turn off the third peristaltic pump 605 and turn off the fourth peristaltic pump 606 to complete the online aseptic sampling operation.

[0060] The embodiments disclosed in the present utility model are preferred embodiments, but are not limited thereto. Ordinary technicians in this field can easily understand the spirit of the present utility model based on the above embodiments and make different extensions and changes. However, as long as they do not deviate from the spirit of the present utility model, they are all within the protection scope of the present utility model.

Claims

1. An online aseptic sampling device for a fermentation process, comprising a bioreactor (1), characterized in that: The online aseptic sampling system also has at least: A sampling assembly (2), one end of which extends below the liquid surface of the bioreactor (1); and A liquid extraction component (3), the liquid extraction component (3) comprising a pinch valve (301) connected to the other end of the sampling component (2) to obtain the liquid to be sampled, and a first three-way connector (303) and a first peristaltic pump (302) connected in sequence to the pinch valve (301); and A backflush assembly (4), the backflush assembly (4) comprising a first gas storage device (404) with a first solenoid valve (402) and a second gas storage device (405) with a second solenoid valve (403), the first solenoid valve (402) and the second solenoid valve (403) being respectively connected to the first three-way connector (303) via a second three-way interface (406); A quantitative component (5), the quantitative component (5) comprising a quantitative tube (502) sequentially connected to the first peristaltic pump (302), the quantitative tube (502) having a liquid path bubble sensor (501); as well as a sample receiving assembly (6), the sample receiving assembly (6) having a third three-way joint (607) configured to be connected to the quantitative tube (502), one interface of the third three-way joint (607) being connected in sequence to a one-way valve (609), a second peristaltic pump (604) and a cleaning liquid bottle (602) to obtain cleaning liquid in the cleaning liquid bottle (602), and another interface of the third three-way joint (607) being connected to a sampling pool (603); and A control device (7), wherein the control device (7) is respectively connected to the clamp valve (301), the first peristaltic pump (302), the first solenoid valve (402), the second solenoid valve (403), the liquid path bubble sensor (501) and the second peristaltic pump (604) to control their opening and closing states.

2. The online aseptic sampling device for fermentation process according to claim 1, characterized in that: The sampling pool (603) is provided with three ports, namely, a liquid inlet, a liquid outlet and an overflow port, wherein the overflow port and the liquid outlet are respectively connected to the waste liquid interface (601) through the third peristaltic pump (605) and the fourth peristaltic pump (606) to discharge the waste liquid.

3. The online aseptic sampling device for fermentation process according to claim 2, characterized in that: The length of the quantitative tube (502) of the quantitative component (5) is changed according to the size of the sampling amount.

4. The online aseptic sampling device for fermentation process according to claim 2, characterized in that: The pinch valve (301) is an electromagnetic pulse pinch valve, and its working state is normally closed (NC).

5. The online aseptic sampling device for fermentation process according to any one of claims 1 to 4, characterized in that: The first gas storage device (404) comprises a first gas container (404-1), and a first gas filter (404-2) and a first gas outlet (404-3) respectively connected to the first gas container (404-1); The second gas storage device (405) comprises a second gas container (405-1), and a second gas filter (405-2) and a second gas outlet (405-3) respectively connected to the second gas container (405-1); Wherein: the first gas filter (404-2) and the second gas filter (405-2) are respectively directly connected to the atmosphere or are the inlet of the second gas storage device (405).

6. The on-line aseptic sampling device for fermentation process according to claim 5, characterized in that: The bioreactor (1), sampling component (2), liquid extraction component (3), backflush component (4), quantitative component (5) and sample receiving component (6) are connected by pipelines, and the connecting pipelines are all silicone pipelines or PU gas pipelines.

7. The online aseptic sampling device for fermentation process according to claim 6, characterized in that: The bioreactor (1) is a culture tank made of glass or stainless steel.

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