Combined deep air inlet device of bioreactor and bioreactor

Abstract

The utility model provides a combined deep air inlet device of a bioreactor and the bioreactor. The combined deep air inlet device comprises an air inlet assembly, an upper air inlet bubbling ring and a lower air inlet microbubble ring, the upper air inlet bubbling ring and the lower air inlet microbubble ring are connected with the air inlet assembly, the upper air inlet bubbling ring is arranged between the upper stirring blade paddle and the middle stirring blade paddle, and the lower air inlet microbubble ring is arranged below the lower stirring blade paddle; the upper air inlet bubbling ring is provided with a first air inlet hole communicated with the air inlet assembly, the lower air inlet bubbling ring is provided with a second air inlet hole communicated with the air inlet assembly, and the first air inlet hole and the second air inlet hole are used for generating bubbles. According to the utility model, the concentration of dissolved oxygen can be greatly improved, and cell damage caused by breakage of bubbles formed in the deep air inlet process in the rising process is reduced.

Description

Technical Field

[0001] The utility model relates to the technical field of bioreactors, in particular to a combined deep air intake device for a bioreactor and a bioreactor. Background Art

[0002] In the production process of foot-and-mouth disease vaccine, dual high-efficiency suspension culture technology (high cell density virus culture, efficient virus expression) is applied to foot-and-mouth disease vaccine production, which can greatly improve the production efficiency of unit volume container, with high virus expression and stable expression process, and can also shorten the production cycle and reduce equipment investment, thereby reducing production costs and improving production efficiency. Therefore, dual high-efficiency suspension culture technology is one of the fastest-growing emerging technologies in the field of animal vaccine production today.

[0003] For the metabolic activities of animal cells, they need to inhale oxygen and expel carbon dioxide. When animal cells breathe, they can only use oxygen molecules dissolved in the culture medium. Oxygen molecules can only be used by animal cells after entering the cells through the cell membrane. Therefore, dissolved oxygen concentration is one of the important factors affecting the growth and metabolism of animal cells in animal cell culture, especially in high cell density culture. Animal cells have different requirements for dissolved oxygen at different growth stages. Appropriate control of dissolved oxygen concentration plays an important role in the process of animal cell culture and virus culture; especially in the high cell density virus culture process, BHK21 cells are in a stable growth stage and consume a lot of oxygen. Meeting the demand for dissolved oxygen often becomes a crucial factor affecting the expression of viral antigens. For high cell density virus culture, in order to maintain an appropriate dissolved oxygen concentration during the culture process, the bioreactor must have a wide range of control capabilities to adjust dissolved oxygen.

[0004] The deep air intake device of the traditional sanitary large-scale bioreactor is a single-inlet bubbling ring device, which is located under the stirring blade of the stirring shaft of the bioreactor. The bubbling ring device is a pipe with several stainless steel semicircular rings with a closed end, and its aperture is between 5mm and 10mm. When culturing cells or viruses in the bioreactor, sterile air and oxygen after sterilization filtration enter the culture solution through the deep air intake device, and the DCS system controls the ventilation volume, stirring speed, and pressure in the tank to achieve oxygen dissolution in the culture solution and the concentration of dissolved oxygen meets the growth and metabolic needs of animal cells. For low-density cell culture, when the oxygen demand is small, the single-inlet deep air intake device of the bioreactor can meet the growth and metabolic needs, but in the high-density cell culture process, when the maximum oxygen demand is large, it is difficult to solve it by the above method: 1. Large ventilation volume and high speed lead to high energy consumption; 2. Large ventilation volume, high air or oxygen flow rate, low oxygen utilization rate, foam is easy to form in the tank, and the bubbles burst during the rising process, causing cell damage.

[0005] The patent document with publication number CN102399690A discloses a dissolved oxygen control system in a high cell density fermentation process, which can well solve the problem of greatly regulating dissolved oxygen in a high cell density fermentation process. It sets a sterile air distributor between the inlet and outlet of the fermentation tank, and sets a fermentation liquid circulation pump, a fermentation liquid bypass control valve, and a fermentation liquid pressure regulating control valve in the distributor between the fermentation tank and the sterile air distributor. Its working mode is: the fermentation liquid is led out of the tank, and the dissolved oxygen concentration of the fermentation liquid is adjusted and the distributor membrane is cleaned on the way of the fermentation liquid being led out, which can greatly increase the dissolved oxygen concentration, realize the regulation of a wide range of dissolved oxygen, and save energy consumption at the same time. However, this patent document is only applicable to an aerobic microbial fermentation process, which is a control system for dissolved oxygen in the fermentation process, and is not applicable to improving the efficiency of dissolved oxygen in the high cell density culture of foot-and-mouth disease virus, because the culture of animal cells is a refined culture operation. Utility Model Content

[0006] In view of the defects in the prior art, the purpose of the utility model is to provide a bioreactor combined deep air intake device and a bioreactor.

[0007] According to the utility model, a combined deep air intake device for a bioreactor comprises: an air intake assembly, an upper air intake bubbling ring and a lower air intake microbubble ring;

[0008] The upper air intake bubbling ring and the lower air intake microbubble ring are connected to the air intake assembly, the upper air intake bubbling ring is arranged between the upper stirring blade and the middle stirring blade, and the lower air intake microbubble ring is arranged below the lower stirring blade;

[0009] The upper air inlet bubbling ring is provided with a first air inlet hole connected to the air inlet assembly, and the lower air inlet microbubble ring is provided with a second air inlet hole connected to the air inlet assembly. The first air inlet hole and the second air inlet hole are used to generate bubbles.

[0010] Preferably, the upper air inlet bubbling ring is a semicircular pipe with closed ends;

[0011] The pipeline is connected to the air intake assembly, and the first air intake hole is connected to the pipeline;

[0012] The first air inlet holes are arranged in plurality, and the plurality of first air inlet holes are evenly distributed on the pipeline.

[0013] Preferably, the lower air inlet microbubble ring is a cylinder;

[0014] An air intake passage is arranged in the cylinder, the air intake passage is communicated with the air intake assembly, and the second air intake hole is communicated with the air intake passage;

[0015] The second air inlet holes are arranged in plurality, and the plurality of second air inlet holes are evenly distributed on the side surface of the cylinder.

[0016] Preferably, the diameter of the first air inlet hole is 5 mm to 10 mm.

[0017] Preferably, the second air inlet hole has a diameter of 5 μm to 10 μm.

[0018] Preferably, the upper air inlet bubbling ring and the lower air inlet microbubble ring are both made of stainless steel.

[0019] Preferably, the air intake assembly includes a gas storage assembly, a proton flowmeter and an air intake sterilizing filter;

[0020] The gas storage assembly is connected to the proton flowmeter via a manual ball valve, and the proton flowmeter is connected to the air intake sterilizing filter via a first one-way valve;

[0021] The air inlet sterilizing filter is connected to the upper air inlet bubbling ring through a second one-way valve, and the air inlet sterilizing filter is connected to the lower air inlet microbubble ring through a third one-way valve.

[0022] Preferably, the manual ball valve and the proton flowmeter are connected via a pressure reducing valve.

[0023] Preferably, the first one-way valve is connected to the air inlet sterilizing filter via a first pneumatic diaphragm valve;

[0024] The air intake sterilizing filter is connected to the second one-way valve via a second pneumatic diaphragm valve, and the air intake sterilizing filter is connected to the third one-way valve via a third pneumatic diaphragm valve.

[0025] Preferably, the gas storage assembly includes a compressed air storage tank and an oxygen storage tank.

[0026] The utility model also provides a bioreactor, comprising the above-mentioned bioreactor combined deep air intake device.

[0027] Compared with the prior art, the utility model has the following beneficial effects:

[0028] 1. The utility model can realize a single air intake mode and a double air intake mode according to the cell density in the culture material; the single air intake mode is a lower air intake microbubble ring deep air intake device, which is mainly suitable for cultures with low cell density, whose oxygen demand is small and the number of cells per unit volume is small; the double air intake mode is a lower air intake microbubble ring device and an upper air intake bubbling ring device that intake air at the same time, which is mainly suitable for cultures with high cell density, whose oxygen demand is large and the number of cells per unit volume is large.

[0029] 2. The deep air intake of the bioreactor of the utility model is simultaneously from the bottom section and the middle section, which effectively avoids the problems of the traditional bioreactor with large single-point deep air intake ventilation, large air or oxygen flow rate, low oxygen utilization rate, easy formation of foam in the tank, and a large number of bubbles bursting during the rising process, causing cell damage.

[0030] 3. The utility model provides a combined deep air intake device for a bioreactor suitable for high cell density virus culture, which can greatly increase the concentration of dissolved oxygen and reduce the damage to cells caused by the bubbles formed in the deep air intake process bursting during the rising process. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Other features, objects and advantages of the present invention will become more apparent by reading the detailed description of non-limiting embodiments with reference to the following drawings:

[0032] Figure 1 It is a schematic diagram of the structure of a bioreactor with a combined deep gas inlet device for the bioreactor;

[0033] Figure 2 is a schematic diagram of the structure of the upper air intake bubble ring;

[0034] Figure 3 Schematic diagram of the structure of the lower air inlet microbubble ring.

[0035] The figure shows:

[0036] Upper air inlet bubble ring 1 Third one-way valve 15

[0037] Lower air inlet micro bubble ring 2 Pneumatic diaphragm valve 16

[0038] Upper stirring blade 3 First check valve 17

[0039] Middle stirring blade 4 Proton flow meter 18

[0040] Lower stirring blade 5 Compressed air storage tank 19

[0041] Stirring shaft 6 Oxygen storage tank 20

[0042] Sampling valve 7 Carbon dioxide storage tank 21

[0043] Tank bottom valve 8 Manual ball valve 22

[0044] Sight glass 9 Pressure reducing valve 23

[0045] Mirror light 10 First pneumatic diaphragm valve 24

[0046] Exhaust gas sterilizing filter 11 Third pneumatic diaphragm valve 25

[0047] Cleaning / steam inlet 12 Second pneumatic diaphragm valve 26

[0048] Air inlet sterilizing filter 13 DO electrode 27

[0049] Second check valve 14 PH electrode 28 DETAILED DESCRIPTION

[0050] The utility model is described in detail below in conjunction with specific embodiments. The following embodiments will help those skilled in the art to further understand the utility model, but do not limit the utility model in any form. It should be pointed out that for those of ordinary skill in the art, several changes and improvements can be made without departing from the concept of the utility model. These all belong to the protection scope of the utility model.

[0051] Embodiment 1:

[0052] like Figures 1 to 3 As shown, this embodiment provides a combined deep air intake device for a bioreactor, comprising: an air intake assembly, an upper air intake bubbling ring 1 and a lower air intake microbubble ring 2; the upper air intake bubbling ring 1 and the lower air intake microbubble ring 2 are connected to the air intake assembly, the upper air intake bubbling ring 1 is arranged between the upper stirring blade 3 and the middle stirring blade 4, and the lower air intake microbubble ring 2 is arranged below the lower stirring blade 5; the upper air intake bubbling ring 1 is provided with a first air intake hole 101 connected to the air intake assembly, and the lower air intake microbubble ring 2 is provided with a second air intake hole 201 connected to the air intake assembly, and the first air intake hole 101 and the second air intake hole 201 are used to generate bubbles. The aperture of the first air intake hole 101 is 5mm to 10mm. The aperture of the second air intake hole 201 is 5μm to 10μm. The upper air intake bubbling ring 1 and the lower air intake microbubble ring 2 are both stainless steel structures.

[0053] The air intake component includes a gas storage component, a proton flowmeter 18 and an air intake sterilizing filter 13; the gas storage component is connected to the proton flowmeter 18 through a manual ball valve 22, and the proton flowmeter 18 is connected to the air intake sterilizing filter 13 through a first one-way valve 17; the air intake sterilizing filter 13 is connected to the upper air intake bubbling ring 1 through a second one-way valve 14, and the air intake sterilizing filter 13 is connected to the lower air intake microbubble ring 2 through a third one-way valve 15.

[0054] The upper air inlet bubbling ring 1 is a semicircular pipe with closed ends; the pipe is connected to the air inlet assembly, and the first air inlet hole 101 is connected to the pipe; there are multiple first air inlet holes 101, and the multiple first air inlet holes 101 are evenly distributed on the pipe. The lower air inlet microbubble ring 2 is a cylinder; an air inlet channel is provided in the cylinder, the air inlet channel is connected to the air inlet assembly, and the second air inlet hole 201 is connected to the air inlet channel; there are multiple second air inlet holes 201, and the multiple second air inlet holes 201 are evenly distributed on the side of the cylinder.

[0055] The manual ball valve 22 is connected to the proton flowmeter 18 via a pressure reducing valve 23. The first one-way valve 17 is connected to the air inlet sterilizing filter 13 via a first pneumatic diaphragm valve 24; the air inlet sterilizing filter 13 is connected to the second one-way valve 14 via a second pneumatic diaphragm valve 26, and the air inlet sterilizing filter 13 is connected to the third one-way valve 15 via a third pneumatic diaphragm valve 25. The gas storage assembly includes a compressed air storage tank 19 and an oxygen storage tank 20. The gas storage assembly also includes a carbon dioxide storage tank 21.

[0056] The combined deep air intake device for bioreactors of this embodiment is applied to a bioreactor with a stirring device, and the upper air intake bubbling ring 1 and the lower air intake microbubble ring 2 are arranged at different positions of the stirring blades of the stirring device. The stirring shaft 6 drives the upper stirring blade 3, the middle stirring blade 4 and the lower stirring blade 5 to rotate.

[0057] This embodiment also provides a bioreactor, comprising the above-mentioned bioreactor combined deep gas inlet device. The bioreactor is also provided with a sampling valve 7, a tank bottom valve 8, a sight glass 9, a sight glass light 10, an exhaust gas sterilizing filter 11, a cleaning / steam inlet 12, a DO electrode 27 and a pH electrode 28.

[0058] The utility model adopts a combined deep air intake device for a bioreactor to solve the problems in the high cell density virus culture process of a traditional large-scale bioreactor, such as large ventilation volume of single-point deep air intake in the bioreactor, large air or oxygen flow rate, low oxygen utilization rate, easy formation of foam in the tank, and cell damage caused by the rupture of a large number of bubbles during the rising process. The utility model achieves the goal of evenly distributing the bubbles generated during the deep air intake process in the bioreactor, increasing the contact area between gas and liquid, improving the efficiency of oxygen dissolving in liquid, and reducing the damage to cells caused by the rupture of bubbles during the rising process.

[0059] The utility model adopts a combined deep air intake device for a bioreactor, thereby solving the problem that a traditional large-scale bioreactor can only have single-point deep air intake, and can realize a single air intake mode and a double air intake mode according to the cell density in the culture material.

[0060] Embodiment 2:

[0061] Those skilled in the art may understand this embodiment as a more specific description of Embodiment 1.

[0062] The present embodiment provides a combined deep gas intake device for a bioreactor suitable for high cell density virus culture, which can significantly increase the concentration of dissolved oxygen and reduce the damage to cells caused by the rupture of bubbles formed during the deep gas intake process during the ascending process.

[0063] The combined deep air intake device of the bioreactor comprises an air intake component, an upper air intake bubbling ring 1 and a lower air intake microbubble ring 2; the upper air intake bubbling ring 1 and the lower air intake microbubble ring 2 are connected to the air intake component, the upper air intake bubbling ring 1 is arranged between the upper stirring blade 3 and the middle stirring blade 4, and the lower air intake microbubble ring 2 is arranged below the lower stirring blade 5; the upper air intake bubbling ring 1 is provided with a first air intake hole 101 connected to the air intake component, and the lower air intake microbubble ring 2 is provided with a second air intake hole 201 connected to the air intake component, and the first air intake hole 101 and the second air intake hole 201 are used to generate bubbles.

[0064] A lower air intake microbubble ring 2 is arranged below the lower stirring blade 5 of the stirring shaft of the bioreactor. The microbubble ring device is a stainless steel cylinder with a plurality of tiny pores, and the pore size is between 5 μm and 10 μm. An upper air intake bubbling ring device is arranged at the middle section of the upper stirring blade 3 and the middle stirring blade 4 of the stirring shaft of the bioreactor. The bubbling ring device is a stainless steel semicircular ring with a plurality of tiny pores, and the two ends of the pipe are closed. The pore size is between 5 mm and 10 mm. The oxygen, clean compressed air and carbon dioxide required for the bioreactor culture process all enter the culture solution through 316L stainless steel pipes, ball valves, pressure reducing valves, proton flowmeters, gas sterilizing filters, one-way valves, lower air intake microbubble ring devices, and upper air intake bubbling ring devices.

[0065] This embodiment needs to meet the use requirements of GMP equipment. The device must be made of 316L stainless steel, have no dead corners, be easy to clean, and be easy to sterilize.

[0066] This embodiment can realize a single air intake mode and a double air intake mode according to the cell density in the culture material. The single air intake mode is a deep air intake device with a lower air intake microbubble ring, which is mainly suitable for cultures with low cell density, whose oxygen demand is small and the number of cells per unit volume is small. Compared with the bubbling ring of a traditional large-scale bioreactor, the bubbles generated by the microbubble ring are small and dense. Under the stirring of the stirring shaft, it is easier to be evenly distributed in the culture solution, and the efficiency of oxygen dissolving in the liquid is better. The double air intake mode is that the lower air intake microbubble ring device and the upper air intake bubbling ring device are simultaneously inlet, which is mainly suitable for cultures with high cell density, whose oxygen demand is large and the number of cells per unit volume is large. Compared with the bubbling ring of a traditional large-scale bioreactor, the lower air intake microbubble ring is located at the bottom of the reactor and the bubbles generated are small and dense, and the upper air intake bubbling device is located in the middle of the reactor, and the bubbles generated are larger than those of the microbubble ring, and are evenly distributed in the culture solution under the stirring of the stirring shaft. Because the bioreactor's deep air intake is simultaneously from the bottom and middle sections, it effectively avoids the problems of traditional bioreactors with large single-point deep air intake, large air or oxygen flow rate, low oxygen utilization rate, easy formation of foam in the tank, and a large number of bubbles bursting during the rising process, causing cell damage.

[0067] It can be seen from the above scheme that this embodiment is an improvement proposed for existing large-scale bioreactor equipment to be suitable for high cell density virus culture. The air intake under the microbubble ring and the air intake on the bubbling ring work together with the stirring of the bioreactor, so that the bubbles generated in the deep air intake process are evenly distributed in the bioreactor, increasing the contact area between the gas and the liquid, improving the efficiency of oxygen dissolving in the liquid, and reducing the damage to the cells caused by the rupture of bubbles during the rising process.

[0068] The combined deep gas inlet device of the bioreactor consists of two parts, namely, an upper gas inlet bubbling ring 1 and a lower gas inlet microbubble ring 2. The aperture of the upper gas inlet bubbling ring 1 is between 5mm and 10mm, and the aperture of the lower gas inlet microbubble ring 2 is between 5μm and 10μm, thereby controlling the size of the bubbles of gas entering the liquid. The upper gas inlet bubbling ring 1 is located in the middle of the upper stirring blade 3 and the middle stirring blade 4. The bubbling ring is a stainless steel semicircular ring with several tiny holes and a closed pipe at both ends, such as Figure 2 The lower air inlet microbubble ring 2 is located at the bottom end of the lower stirring blade 5 of the bioreactor, and the microbubble ring is a stainless steel cylinder with a plurality of tiny pores, such as Figure 3 The device can realize single air inlet mode and double air inlet mode according to the cell density in the culture material.

[0069] The single air intake mode is suitable for low cell density virus culture. Clean compressed air and oxygen are mainly supplied through the manual ball valve 22, the pressure reducing valve 23, the proton flowmeter 18, the first one-way valve 17, the first pneumatic diaphragm valve 24, the air intake sterilizing filter 13, the third pneumatic diaphragm valve 25, the third one-way valve 15, and the lower air intake microbubble ring 2. Together with the upper stirring blade 3, the middle stirring blade 4, and the lower stirring blade 5 of the bioreactor rotating with the stirring shaft 6, single-point deep air intake is performed to meet the dissolved oxygen demand of low cell density virus culture.

[0070] The double air intake mode is suitable for high cell density virus culture. Clean compressed air and oxygen are mainly passed through the manual ball valve 22, the pressure reducing valve 23, the proton flowmeter 18, the first one-way valve 17, the first pneumatic diaphragm valve 24, the air intake sterilizing filter 13, the third pneumatic diaphragm valve 25, the second pneumatic diaphragm valve 26, the second one-way valve 14, the third one-way valve 15, the upper air intake bubbling ring 1, the lower air intake microbubble ring 2, and the upper stirring blade 3, the middle stirring blade 4, and the lower stirring blade 5 of the bioreactor rotating with the stirring shaft 6 to make the bubbles generated in the deep air intake process evenly distributed in the bioreactor, increase the contact area between the gas and the liquid, improve the efficiency of oxygen dissolving in the liquid, and reduce the damage to the cells caused by the rupture of bubbles during the rising process.

[0071] The ventilation volume and stirring speed control of the combined deep-layer aeration device of the bioreactor are controlled by the DCS control system of the existing bioreactor. The DO electrode 27 (dissolved oxygen electrode) transmits the dissolved oxygen concentration value measured in the culture solution to the DCS control system, and the DCS system controls the proton flowmeter 18, the third pneumatic diaphragm valve 25, and the second pneumatic diaphragm valve 26 to control the ventilation volume of clean compressed air and oxygen, and control the stirring speed of the stirring shaft 6, thereby realizing the dissolved oxygen control of the culture solution in the bioreactor.

[0072] The utility model can greatly increase the concentration of dissolved oxygen and reduce the damage to cells caused by the rupture of bubbles formed in the deep air intake process during the ascending process.

[0073] In the description of the present application, it should be understood that the terms "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present application.

[0074] The above describes the specific embodiments of the present invention. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art can make various changes or modifications within the scope of the claims, which does not affect the essence of the present invention. In the absence of conflict, the embodiments of the present application and the features in the embodiments can be combined with each other at will.

Claims

1. A combined deep gas inlet device for a bioreactor, characterized in that: include: An air intake assembly, an upper air intake bubbling ring (1) and a lower air intake microbubble ring (2); The upper air intake bubbling ring (1) and the lower air intake micro-bubble ring (2) are connected to the air intake assembly, the upper air intake bubbling ring (1) is arranged between the upper stirring blade (3) and the middle stirring blade (4), and the lower air intake micro-bubble ring (2) is arranged below the lower stirring blade (5); The upper air inlet bubbling ring (1) is provided with a first air inlet hole (101) connected to the air inlet assembly, and the lower air inlet microbubble ring (2) is provided with a second air inlet hole (201) connected to the air inlet assembly. The first air inlet hole (101) and the second air inlet hole (201) are used to generate bubbles.

2. The combined deep gas inlet device for bioreactor according to claim 1, characterized in that: The upper air inlet bubbling ring (1) is a semicircular pipe with closed ends; The pipeline is connected to the air intake assembly, and the first air intake hole (101) is connected to the pipeline; The first air inlet holes (101) are arranged in plurality, and the plurality of first air inlet holes (101) are evenly distributed on the pipeline.

3. The combined deep gas inlet device for bioreactor according to claim 1, characterized in that: The lower air inlet microbubble ring (2) is a cylinder; An air intake channel is arranged in the cylinder, the air intake channel is connected to the air intake assembly, and the second air intake hole (201) is connected to the air intake channel; The second air inlet holes (201) are arranged in plurality, and the plurality of second air inlet holes (201) are evenly distributed on the side surface of the cylinder.

4. The combined deep gas inlet device for bioreactor according to claim 1, characterized in that: The diameter of the first air inlet hole (101) is 5 mm to 10 mm.

5. The combined deep gas inlet device for bioreactor according to claim 1, characterized in that: The aperture of the second air inlet hole (201) is 5 μm to 10 μm.

6. The combined deep gas inlet device for bioreactor according to claim 1, characterized in that: The upper air inlet bubbling ring (1) and the lower air inlet micro-bubble ring (2) are both made of stainless steel.

7. The combined deep gas inlet device for bioreactor according to claim 1, characterized in that: The air intake assembly comprises a gas storage assembly, a proton flow meter (18) and an air intake sterilizing filter (13); The gas storage assembly is connected to the proton flowmeter (18) via a manual ball valve (22), and the proton flowmeter (18) is connected to the air intake sterilizing filter (13) via a first one-way valve (17); The air intake sterilizing filter (13) is connected to the upper air intake bubbling ring (1) via a second one-way valve (14), and the air intake sterilizing filter (13) is connected to the lower air intake microbubble ring (2) via a third one-way valve (15).

8. The combined deep gas inlet device for bioreactor according to claim 7, characterized in that: The manual ball valve (22) and the proton flowmeter (18) are connected via a pressure reducing valve (23).

9. The combined deep gas inlet device for a bioreactor according to claim 7, characterized in that: The first one-way valve (17) is connected to the air inlet sterilizing filter (13) via a first pneumatic diaphragm valve (24); The air intake sterilizing filter (13) and the second one-way valve (14) are connected via a second pneumatic diaphragm valve (26), and the air intake sterilizing filter (13) and the third one-way valve (15) are connected via a third pneumatic diaphragm valve (25).

10. A bioreactor, characterized in that: A bioreactor combined deep gas inlet device comprising any one of claims 1 to 9.

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