Bypass monitoring and cleaning device for cell culture and cell reaction system

Abstract

The utility model discloses a bypass monitoring and cleaning device for cell culture and a cell reaction system, and belongs to the technical field of cell culture devices.The bypass monitoring and cleaning device comprises an on-line monitoring assembly, a connecting pipeline and a cleaning assembly, the on-line monitoring assembly is communicated with the cleaning assembly through the connecting pipeline, and the cleaning assembly is communicated with the on-line monitoring assembly through the connecting pipeline. The online monitoring assembly comprises a transparent flow cell and a bypass electrode, the flow cell is connected with a liquid inlet pipeline and a liquid outlet pipeline, the bypass electrode comprises an electrode tip, and the electrode tip is arranged in the flow cell. According to the bypass monitoring and cleaning device for cell culture and the cell reaction system, whether pollution exists on the bypass electrode or not can be visually checked, and pollution of the bypass electrode can be cleaned.

Description

Technical Field

[0001] This utility model relates to the field of cell culture device technology, and in particular to an online electrode cleaning device for cell culture. Background Technology

[0002] Cell culture refers to a technique that involves taking cells from in vivo tissues, simulating the in vivo environment, and allowing them to grow, multiply, and maintain their structure and function under sterile, appropriate temperature, pH, and nutritional conditions. In biopharmaceuticals, cell culture technology is widely used in the production of recombinant protein drugs, monoclonal antibodies, vaccines, cell and gene therapies, and other biological agents, making significant contributions to human health.

[0003] When using reactors for cell culture, various electrodes are installed in the reactor tanks for monitoring parameters such as pH, dissolved oxygen (DO), optical density (OD), cell density (CD), and even cell expression and metabolites. However, electrodes may be damaged or malfunction during tank sterilization or electrode sterilization, or even during long-term cell culture. Currently, there are three main cell culture methods: adherent culture, suspension culture, and immobilized culture. In adherent or immobilized culture, cells typically need to adhere to or be fixed to the culture medium. However, during inoculation or culture, cells may still adhere to the electrodes used for online monitoring, and cell debris adsorption and electrode contamination are unavoidable, affecting the precise control of the electrodes during cell culture. This phenomenon becomes more severe over time. Suspension culture also faces the possibility of electrode malfunction or contamination, especially with the development and widespread adoption of continuous culture processes, which significantly extends the cell culture cycle and increases the risk of electrode malfunction and contamination. Utility Model Content

[0004] To address the problem that monitoring electrodes in existing technologies are easily contaminated and difficult to observe and handle, this invention proposes an online electrode cleaning device for cell culture.

[0005] The specific technical solution is as follows:

[0006] A bypass monitoring and cleaning device for cell culture, characterized in that it comprises: an online monitoring component, a connecting pipeline, and a cleaning component, wherein the online monitoring component and the cleaning component are connected through the connecting pipeline, the online monitoring component includes a transparent flow cell and a bypass electrode, the flow cell is connected to an inlet pipeline and an outlet pipeline, and the bypass electrode includes an electrode head disposed within the flow cell.

[0007] Furthermore, the online monitoring component also includes a flow cell housing and a seal, with the top of the flow cell connected to the bottom of the flow cell housing via the seal, which divides the flow cell and the flow cell housing into two independent cavities.

[0008] Furthermore, the bypass electrode also includes an electrode connector and an electrode rod arranged from top to bottom. The electrode connector is located at the top of the flow cell housing, the electrode rod is located inside the flow cell housing and passes through the seal, and the electrode head is located at the bottom of the electrode rod.

[0009] Furthermore, a third quick connector is provided at the bottom of the flow pool, the connecting pipeline includes a silicone tube, one end of the silicone tube is connected to a fourth quick connector, an eighth tube clamp is provided at the fourth quick connector, and the third quick connector is connected to the fourth quick connector.

[0010] Furthermore, the silicone tube includes multiple branch lines connected to quick-connect fittings.

[0011] Furthermore, the cleaning assembly includes multiple screw-in syringes.

[0012] Furthermore, the flow cell is made of a flexible material.

[0013] Furthermore, the flow pool is connected to the respirator, and the respirator is equipped with a first tube clamp.

[0014] A cell reaction system, characterized in that it comprises: a bioreactor and a bypass monitoring and cleaning device for cell culture as described in any one of the claims, wherein the bioreactor is connected to the flow cell via the inlet pipe and the outlet pipe.

[0015] Furthermore, the inlet pipe is provided with a first quick connector at one end, the outlet pipe is provided with a second quick connector at one end, the inlet pipe is provided with a second pipe clamp, and the outlet pipe is provided with a third pipe clamp. The bioreactor is connected to the first quick connector and the second quick connector respectively.

[0016] The above technical solution has the following advantages or technical effects:

[0017] 1. Based on the original online monitoring of bioreactors, this utility model adds bypass monitoring technology. By observing the surface of the bypass electrode through a transparent flow cell, the surface condition of the bypass electrode can be detected in time when the liquid volume in the flow cell is small.

[0018] 2. The flow cell is made of a flexible, transparent material, which allows for easy observation and also enables manual crushing or removal of contaminants from the bypass electrode surface by kneading.

[0019] 3. The screw-type syringe of the cleaning component can perform a combination of operations such as emptying, cleaning and digestion to remove cells or cell debris adhering to the bypass electrode.

[0020] 4. When an electrode malfunctions in the bioreactor, this invention can install a normal electrode and replace the function of the malfunctioning electrode through bypass monitoring. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of the structure of the device of this utility model;

[0022] Figure 2 This is a schematic diagram of the structure of the online monitoring component of this utility model;

[0023] Figure 3 This is a schematic diagram of the connecting pipeline of this utility model;

[0024] Figure 4 This is a schematic diagram of the cleaning component of this utility model;

[0025] Figure 5 This is a schematic diagram showing the connection between the online monitoring component of this utility model and the bioreactor;

[0026] The attached diagram is labeled as follows: 1-Online monitoring component, 101-Flow cell housing, 102-Seal, 103-Flow cell, 104-Inlet pipe, 105-Outlet pipe, 106-Third quick connector, 107-First clamp, 108-Second clamp, 109-Third clamp, 110-Breathe, 2-Connecting pipe, 201-Fourth quick connector, 202-Fifth quick connector, 203-Sixth quick connector, 204-Seventh quick connector 205-Eighth quick connector, 206-Silicone tubing, 207-Fourth clamp, 208-Fifth clamp, 209-Sixth clamp, 210-Seventh clamp, 211-Eighth clamp, 3-Cleaning assembly, 301-First screw-in syringe, 302-Second screw-in syringe, 303-Third screw-in syringe, 304-Fourth screw-in syringe, 4-Bypass electrode, 401-Electrode connector, 402-Electrode rod, 403-Electrode head. Detailed Implementation

[0027] To make the technical solution of this utility model clearer, the present utility model will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0028] like Figure 1As shown, an online electrode cleaning device for cell culture includes an online monitoring component 1 and a cleaning component 3 connected by a connecting pipe 2. The online monitoring component 1 includes a transparent flow cell 103 and a bypass electrode 4. The flow cell 103 is connected to an inlet pipe 104 and an outlet pipe 105. The bypass electrode 4 includes an electrode head 403, which is disposed within the flow cell 103. The flow cell 103, the connecting pipe 2, and the screw-type syringe are a ready-to-use, disposable, sterile system. The flow path between the cleaning device and the bioreactor, as well as the solution circulating within the cleaning device and the bioreactor, are all sterilized or disinfected.

[0029] When an electrode in the bioreactor malfunctions, the online monitoring component 1 provides bypass monitoring for the bioreactor, replacing the function of the faulty electrode with a bypass electrode 4 for online monitoring. It monitors at least one of the following during the culture process: pH, dissolved oxygen, optical density, cell density, cell expression, and metabolites. The inlet pipe 104 of the online monitoring component 1, with a first quick connector (with a plug), connects to a pre-installed bioreactor harvest pipe. The outlet pipe 105 of the online monitoring component 1, with a second quick connector (with a plug), connects to a pre-installed bioreactor return pipe. The flow cell 103 is made of a transparent material, such as a transparent plastic, providing a transparent window to observe the surface condition of the bypass electrode 4 within the online monitoring component 1 and confirm the severity of contamination. When the bypass electrode 4 is contaminated, the online monitoring control corresponding to the bypass electrode 4 requiring cleaning is stopped. The online control cleaning component 3 injects cleaning solution and digestion solution into the flow cell through the connecting pipe 2, thus completing the cleaning of the contaminated bypass electrode 4.

[0030] like Figure 2 As shown, the online monitoring component 1 also includes a flow cell housing 101 and a sealing element 102. The top of the flow cell 103 is connected to the bottom of the flow cell housing 101 via the sealing element 102. The sealing element 102 seals the fixed position of the flow cell housing 101 and the flow cell 103, dividing the flow cell 103 and the flow cell housing 101 into two independent cavities. The lower cavity of the flow cell 103 provides an environment for the online monitoring of the bypass electrode 4 and the flow of the solution. The bypass electrode 4 also includes an electrode connector 401 and an electrode rod 402 arranged from top to bottom. The electrode connector 401 is located at the top of the flow cell housing 101, and the electrode rod 402 is located inside the flow cell housing 101 and passes through the sealing element 102. The electrode head 403 is located at the bottom of the electrode rod 402.

[0031] The inlet pipe 104 is equipped with a first quick connector at one end, and the outlet pipe 105 is equipped with a second quick connector at one end. A second clamp 108 is installed on the inlet pipe 104, and a third clamp 109 is installed on the outlet pipe 105. A third quick connector 106 with a plug is located at the bottom of the flow-through tank 103. The inlet pipe 104 and outlet pipe 105 are connected to the bioreactor, serving as solution inlet and outlet channels for the flow-through tank 103. The third quick connector 106 serves as the inlet for the cleaning component 3.

[0032] like Figure 3 As shown, the connecting pipe 2 includes a silicone tube 206, which is connected to a fourth quick connector 201. An eighth clamp 211 is located at the fourth quick connector 201. A third quick connector 106 is connected to the fourth quick connector 201. The silicone tube 206 includes multiple branch pipes connected to quick connectors. These branches are respectively connected to a fifth quick connector 202, a sixth quick connector 203, a seventh quick connector 204, and an eighth quick connector 205. A fourth clamp 207 is located at the fifth quick connector 202, a fifth clamp 208 at the sixth quick connector 203, a sixth clamp 209 at the seventh quick connector 204, and a seventh clamp 210 at the eighth quick connector 205. By configuring the silicone tube 206 as a multi-branch pipe and installing clamps on each branch pipe, the shut-off of the pipes can be selectively controlled, providing cleaning and digesting solutions to the flow tank 103.

[0033] like Figure 4 As shown, the cleaning assembly 3 includes multiple screw-type syringes, specifically a first screw-type syringe 301, a second screw-type syringe 302, a third screw-type syringe 303, and a fourth screw-type syringe 304. The first screw-type syringe 301 is connected to a fifth quick connector 202, the second screw-type syringe 302 is connected to a sixth quick connector 203, the third screw-type syringe 303 is connected to a seventh quick connector 204, and the fourth screw-type syringe 304 is connected to an eighth quick connector 205. The different screw-type syringes provide different functions and do not interfere with each other, collectively providing online cleaning functions, including emptying, cleaning, and digestion, to remove cells or cell debris adhering to the bypass electrode 4.

[0034] The flow cell 103 is made of a transparent, flexible material that can be kneaded to crush or remove contaminants (usually aggregates of large cell debris) from the surface of the bypass electrode 4. The flow cell 103 is connected to the respirator 110, which is equipped with a first tube clamp 107. By controlling the shut-off of the respirator 110, the air pressure inside the online monitoring component 1 is kept constant, and external contaminants are prevented from entering the cavity.

[0035] like Figure 5As shown, a cell reaction system includes a bioreactor and a bypass monitoring and cleaning device for cell culture. The bioreactor is connected to a flow cell 103 via an inlet pipe 104 and an outlet pipe 105. The inlet pipe 104 is provided with a first quick connector at one end, and the outlet pipe 105 is provided with a second quick connector at one end. The inlet pipe 104 is provided with a second clamp 108, and the outlet pipe 105 is provided with a third clamp 109. The bioreactor is connected to the first quick connector and the second quick connector respectively. A circulation pump is also connected between the second quick connector and the bioreactor.

[0036] In use, the bypass monitoring and cleaning device, which is equipped with bypass electrode 4 and has been sterilized, is fixed on or next to the bioreactor (such as the Scale-X™hydro R&D fixed bed bioreactor). The inlet pipe 104 with the first quick connector (with plug) of the online monitoring component 1 and the reserved bioreactor harvest pipe are connected by a sterile connector or in a biosafety cabinet. The outlet pipe 105 with the second quick connector (with plug) of the online monitoring component 1 and the reserved bioreactor return pipe are connected. The circulation pump between the outlet pipe 105 and the bioreactor return pipe is turned on. The electrode cable and electrode connector 401 are connected.

[0037] The surface condition of the bypass electrode 4 can be observed through the transparent window of the flow cell 103 to confirm the severity of contamination. If there are obvious cell clusters contaminating the electrode head 403, the flexible plastic shell of the flow cell 103 can be kneaded to crush or remove the contaminants on the surface of the bypass electrode 4, allowing them to flow away with the liquid.

[0038] The cleaning process of cleaning component 3 for flow cell 103 is as follows:

[0039] (1) Stop the monitoring and control corresponding to the bypass electrode 4 that needs to be cleaned, close the second clamp 108 and open the first clamp 107, adjust the angle of the flow cell 103 appropriately, and after the liquid in the chamber is drained by the circulation pump, close the circulation pump and close the third clamp 109.

[0040] (2) In the biosafety cabinet, add sterile cleaning solution (such as PBS or Hanks buffer solution), digestion solution (such as 2.5 g / L trypsin + 0.2 g / L EDTA solution) and cleaning solution (such as PBS or Hanks buffer solution) to the first screw-type syringe 301, the second screw-type syringe 302 and the third screw-type syringe 303 respectively. The fourth screw-type syringe 304 is ready for use.

[0041] (3) Aseptically connect the fourth quick connector 201 of the connecting pipe 2 to the third quick connector 106 of the online monitoring component 1, and then aseptically connect the fifth quick connector 202 to the first screw syringe 301, the sixth quick connector 203 to the second screw syringe 302, the sixth quick connector 207 to the third screw syringe 303, and the eighth quick connector 205 for later use.

[0042] (4) Open the fourth clamp 207, inject an appropriate amount of cleaning solution (such as PBS or Hanks buffer solution) into the flow cell 103 using the first screw syringe 301, blow and rinse 1-10 times, repeatedly spray the electrode head 403 and rinse the inner cavity of the flow cell 103, then drain the cleaning solution and close the fourth clamp 207.

[0043] (5) Open the fifth clamp 208, inject an appropriate amount of digestion solution (such as 2.5g / L trypsin + 0.2g / L EDTA solution) into the flow cell 103 using the second screw syringe 302, let it stand for 1-3 minutes, blow and swirl 1-10 times, spray the electrode head 403 repeatedly and rinse the inner cavity of the flow cell 103, then drain the digestion solution and close the fifth clamp 208;

[0044] (6) Open the sixth clamp 209, inject an appropriate amount of cleaning solution (such as PBS or Hanks buffer solution) into the flow cell 103 using the third screw syringe 303, blow and rinse 1-10 times, repeatedly spray the electrode head 403 and rinse the inner cavity of the flow cell 103, then drain the cleaning solution and close the sixth clamp 209 and the first clamp 107.

[0045] (7) Disconnect the third quick connector 106 and the fourth quick connector 201 aseptically, and seal the third connector 106 with a sterile plug. Or, after closing the eighth tube clamp 211, cut the silicone tube 206;

[0046] (8) Open the second clamp 108 and the third clamp 109 in sequence, run the circulation pump for a period of time, and after the liquid in the reactor is filled with the flow tank 103, restart the monitoring and control corresponding to the bypass electrode 4 that has been cleaned.

[0047] The embodiments described above are merely illustrative of several implementations of this utility model, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of this utility model patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this utility model, and these all fall within the protection scope of this utility model. Therefore, the protection scope of this utility model patent should be determined by the appended claims.

Claims

1. A bypass monitoring and cleaning device for cell culture, characterized in that, include: The online monitoring component (1), the connecting pipe (2), and the cleaning component (3) are connected through the connecting pipe (2). The online monitoring component (1) includes a transparent flow cell (103) and a bypass electrode (4). The flow cell (103) is connected to an inlet pipe (104) and an outlet pipe (105). The bypass electrode (4) includes an electrode head (403) which is disposed in the flow cell (103).

2. The bypass monitoring and cleaning device for cell culture according to claim 1, characterized in that, The online monitoring component (1) further includes a flow cell housing (101) and a seal (102). The top of the flow cell (103) is connected to the bottom of the flow cell housing (101) through the seal (102). The seal (102) divides the flow cell (103) and the flow cell housing (101) into two independent cavities.

3. The bypass monitoring and cleaning device for cell culture according to claim 2, characterized in that, The bypass electrode (4) also includes an electrode connector (401) and an electrode rod (402) arranged from top to bottom. The electrode connector (401) is located at the top of the flow cell housing (101), the electrode rod (402) is located inside the flow cell housing (101) and passes through the seal (102), and the electrode head (403) is located at the bottom of the electrode rod (402).

4. The bypass monitoring and cleaning device for cell culture according to claim 1, characterized in that, The bottom of the flow pool (103) is provided with a third quick connector (106), the connecting pipe (2) includes a silicone tube (206), one end of the silicone tube (206) is connected to a fourth quick connector (201), an eighth pipe clamp (211) is provided at the fourth quick connector (201), and the third quick connector (106) is connected to the fourth quick connector (201).

5. The bypass monitoring and cleaning device for cell culture according to claim 4, characterized in that, The silicone tube (206) includes multiple branch tubes connected to quick-connect fittings.

6. The bypass monitoring and cleaning device for cell culture according to claim 1, characterized in that, The cleaning assembly (3) includes multiple screw-in syringes.

7. The bypass monitoring and cleaning device for cell culture according to claim 1, characterized in that, The flow cell (103) is made of a flexible material.

8. The bypass monitoring and cleaning device for cell culture according to claim 1, characterized in that, The flow pool (103) is connected to the respirator (110), and the respirator (110) is provided with a first tube clamp (107).

9. A cell reaction system, characterized in that, include: The bioreactor and the bypass monitoring and cleaning device for cell culture according to any one of claims 1 to 8, wherein the bioreactor is connected to the flow cell (103) via the inlet pipe (104) and the outlet pipe (105).

10. A cell reaction system according to claim 9, characterized in that, The inlet pipe (104) is provided with a first quick connector at its end, the outlet pipe (105) is provided with a second quick connector at its end, the inlet pipe (104) is provided with a second pipe clamp (108), the outlet pipe (105) is provided with a third pipe clamp (109), and the bioreactor is connected to the first quick connector and the second quick connector respectively.

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