CN108504575A - A kind of novel zooblast suspension culture magnetic agitation system - Google Patents

Abstract

The invention discloses a novel magnetic stirring system for animal cell suspension culture, comprising a tank body, a top stirring system arranged above the tank body and a bottom stirring system arranged below the tank body, the top stirring system being the main stirring system , the bottom stirring system is an auxiliary stirring system, the top stirring system includes a power drive device and a top stirring paddle driven by the power drive device, the top stirring paddle goes deep into the inside of the tank, and the bottom stirring system includes a power A driving device and a bottom stirring paddle driven by the power driving device, the bottom stirring paddle is arranged at the bottom of the tank body. The invention solves the problem that there is a gap between the mechanical fits of ordinary mechanical stirring, which is easy to cause the culture solution to be contaminated with bacteria; the double stirring mechanism effectively alleviates the radial runout of the stirring shaft, reduces the shear force borne by the cells, and effectively improves the The mixing uniformity of the culture solution in the reactor; the problem of sedimentation at the bottom is solved; the influence of the magnetic field on the reactor is negligible.

Description

A New Magnetic Stirring System for Animal Cell Suspension Culture

technical field

The invention relates to the technical field of animal cell suspension culture reactors, in particular to a novel magnetic stirring system for animal cell suspension culture.

Background technique

With the outbreak of animal infectious diseases such as bird flu and foot-and-mouth disease, the market demand for veterinary vaccines has increased significantly. As a key technology for the preparation of biological products, large-scale animal cell culture has become an important link in industrial production. The bioreactor is the core equipment to maintain the growth and reproduction of cells in vitro. The cells need enough water, oxygen and other nutrients during the culture process. Therefore, a stirring system is needed to stir the cells and the nutrient solution to mix them evenly. However, the cell culture process can Withstands very low shear forces. At present, ordinary mechanically stirred bioreactors are the most widely used in animal cell culture bioreactors. Ordinary mechanical stirring device, through the action of the stirrer, the cells and nutrients are evenly distributed in the culture medium, so that the nutrients can be fully utilized by the cells, and the gas-liquid contact surface is enlarged, which is conducive to the transfer of oxygen, and can obtain good mixing and mass transfer effects . In an ordinary mechanically stirred bioreactor, the stirrer, the stirring shaft and the transmission device are respectively located inside and outside the tank, and the transmission shaft runs through the inside and outside of the tank, and the gap between the shaft and the tank is generally sealed by a mechanical seal. Mechanical seals are generally reliable and have a long service life, but they have the following disadvantages: (1) There is a gap in the mechanical fit, which is easy to cause the culture solution to be contaminated with bacteria; (2) The maintenance is complicated and the maintenance cost is high.

The function of the magnetic drive stirring equipment and the ordinary mechanical stirring reactor is basically the same except for the power transmission mode. However, the magnetic stirring is completely sealed and pollution-free, which fundamentally solves the problem that there is a gap in the cooperation of ordinary mechanical stirring machinery, which is easy to cause the culture liquid to be contaminated with bacteria, and it is easy to maintain and low in cost. At present, medium and small magnetic drive stirring equipment is widely used and the technology is very mature. With the increasing demand for veterinary vaccines and the continuous expansion of production scale, the scale of magnetic stirring reactors is limited due to some inevitable defects in the development of magnetic stirring. It is necessary to solve the problem technically and develop a new type of magnetic stirrer for large-scale reactors. Magnetic stirring has the advantages of full sealing and simple maintenance, but it also has the following disadvantages: (1) In the development and design of large-volume magnetic stirrers, the stirring shaft is generally long and heavy, and the stirring shaft will produce diameter when it is working. (2) The torque provided by the magnetic stirring is relatively small, and the large-capacity reactor needs a relatively high speed to achieve the stirring effect, and the formed shear force will also increase with the speed; (3) The large-volume single-bottom magnetic stirring requires a relatively Large torque requires strong magnetic force, and long-term strong magnetic field will have adverse effects on cell growth.

Contents of the invention

The present invention is aimed at the optimization of the bioreactor stirring system for animal cell suspension culture, and aims to overcome the problems of common mechanically stirred reactors with gaps in the mechanical fit, easy to cause bacterial contamination of the culture solution, complicated maintenance and high maintenance costs, and ease the application of magnetic stirring In large-volume reactors, the stirring shaft is prone to radial runout, and the stirring torque is insufficient. Large-capacity reactors require a slightly higher speed, resulting in excessive shearing force. It is proposed to focus on top magnetic stirring and bottom magnetic stirring. A new magnetic reactor stirring system for animal cell suspension culture is supplemented.

Technical scheme of the present invention is as follows:

A new type of magnetic stirring system for animal cell suspension culture, comprising a tank body, a top stirring system arranged above the tank body and a bottom stirring system arranged below the tank body, the top stirring system is the main stirring system, and the bottom stirring system The stirring system is an auxiliary stirring system. The top stirring system includes a power drive device and a top stirring paddle driven by the power drive device. The top stirring paddle goes deep into the inside of the tank body. The bottom stirring system includes a power drive device and A bottom stirring paddle driven by a power drive device, and the bottom stirring paddle is arranged at the bottom of the tank body.

As a further improvement of the present invention, the power drive device includes a driver, an outer magnetic rotor, an inner magnetic rotor, a stirring shaft, and a spacer sleeve. The inner magnetic rotor is driven to rotate by the outer magnetic rotor through magnetic coupling, and the stirring shaft and the inner magnetic The rotors are connected and driven to rotate by the inner magnetic rotor, and the isolation sleeve is arranged on the top or bottom of the tank to isolate the inner magnetic rotor and the outer magnetic rotor.

As a further improvement of the present invention, the power drive device used to drive the top stirring paddle is a top power drive device, and the top power drive device includes a top stirring driver, a top stirring outer magnetic rotor, a top stirring isolation sleeve, and a top stirring inner magnetic rotor , a stirring rod, a top stirring paddle, the stirring rod is used to connect the top stirring inner magnetic rotor and the top stirring paddle, and the top stirring paddle goes deep into the tank body to stir the liquid in the tank body.

As a further improvement of the present invention, the power drive device used to drive the bottom stirring paddle is a bottom power drive device, and the bottom power drive device includes a bottom stirring paddle, a bottom stirring inner magnetic rotor, a bottom stirring isolation sleeve, and a bottom stirring outer magnetic rotor 1. Bottom stirring driver, the bottom stirring paddle is directly connected with the bottom stirring inner magnetic rotor, and the bottom stirring inner magnetic rotor drives it to rotate to stir the liquid at the bottom of the tank.

As a further improvement of the present invention, the top stirring impeller is the main stirring impeller, and the four-blade stirring impeller is adopted as an axial-flow stirring impeller with spiral conical curved blades.

As a further improvement of the present invention, the bottom stirring paddle is an auxiliary stirring paddle, and an oblique three-bladed paddle is used.

The beneficial effects of the present invention are as follows:

The present invention relates to a new type of large-scale animal cell suspension culture reactor magnetic stirring system with top magnetic stirring as main and bottom magnetic stirring as supplementary. Its advantages are as follows:

(1) The use of magnetic stirring instead of ordinary mechanical stirring solves the problem that there is a gap between the mechanical fits of ordinary mechanical stirring and the culture solution is easily contaminated with bacteria. The reactor is fully sealed and pollution-free, and it is easy to disassemble and maintain.

(2) The single stirring mechanism is replaced by double stirring mechanism, the load borne by each is greatly reduced, which effectively alleviates the radial runout problem of the stirring shaft, reduces the shear force on the cells, and effectively improves the culture medium in the reactor. uniformity of mixing.

(3) General large-scale reactors are mainly top-stirred, and the bottom of the top-stirred reactor cannot be fully stirred, and precipitation is easy to occur; the top magnetic stirring is mainly used, and the bottom magnetic stirring is supplemented, which solves the problem of bottom sedimentation.

(4) The present invention uses the top magnetic stirring as the main stirring method, and the bottom magnetic stirring is the stirring method for auxiliary stirring. The top stirring paddle penetrates into a suitable position inside the tank body through the stirring rod, so the influence of the top magnetic field on the reactor can be ignored , In addition, the bottom is auxiliary stirring, setting a reasonable stirring time interval, does not need to act for a long time, experiments show that the influence of a short-term magnetic field on cell growth is also negligible.

Description of drawings

Figure 1 is a schematic structural diagram of a novel large-scale animal cell suspension culture bioreactor.

Fig. 2 is a schematic diagram of the magnetic drive structure.

Fig. 3 is an exploded schematic view of the bottom magnetic stirring device.

Fig. 4 is a diagram of four wide-blade stirring paddles.

Figure 5 is a diagram of an oblique three-blade stirring paddle.

In the figure: 1-top stirring drive; 2-top stirring outer magnetic rotor; 3. Top stirring spacer; 4-top stirring inner magnetic rotor; 5-tank; 6-stirring rod; 7-top stirring paddle; 8- Bottom stirring paddle; 9‐bottom stirring inner magnetic rotor; 10‐bottom stirring spacer; 11‐bottom stirring outer magnetic rotor; 12‐bottom stirring driver; 13‐driver; 14‐outer magnetic rotor; Stirring shaft; 17‐inner magnetic rotor.

Detailed ways

In order to make the purpose and features of the present invention more comprehensible, the specific implementation manners of the present invention will be further described below in conjunction with the accompanying drawings.

Fig. 1 is a structural schematic diagram of a new type of large-scale animal cell suspension culture bioreactor with top magnetic stirring as main and bottom magnetic stirring as supplementary in the present invention.

Figure 1 includes the top stirring driver 1, which provides power for the entire main stirring, that is, the top stirring system; the top stirring outer magnetic rotor 2, is connected with the top stirring isolation sleeve 3 and the top stirring driver 1, and the electromagnetic energy is converted into mechanical energy by the top stirring driver 1 Drive the top-stirring outer magnetic rotor 2 to rotate; the top-stirring isolation sleeve 3 is used to isolate the top-stirring outer magnetic rotor 2 and the top-stirring inner magnetic rotor 4, and is directly welded to the top of the reactor tank 5, so that the inside and outside of the tank 5 are completely Isolation makes the interior of the reactor fully sealed and pollution-free; the top stirring inner magnetic rotor 4 is driven by the top stirring outer magnetic rotor 2 through magnetic coupling to rotate; the stirring rod 6 is used to connect the top stirring inner magnetic rotor 4 and the top stirring Paddle 7, the top stirring paddle 7 goes deep into the suitable position of tank body 5 to stir the liquid in the tank body 5; Bottom stirring driver 12, provides power for the whole auxiliary stirring that is the bottom stirring system; Bottom stirring external magnetic rotor 11, and bottom stirring The stirring isolation sleeve 10 is connected with the bottom stirring driver 12, and the bottom stirring outer magnetic rotor 11 is driven to rotate by the bottom stirring driver 12 by converting electromagnetic energy into mechanical energy; the bottom stirring isolation sleeve 10 is used to isolate the bottom stirring outer magnetic rotor 11 from the bottom stirring inner The magnetic rotor 9 is directly welded to the bottom of the reactor tank 5, so that the inside of the tank 5 is completely isolated from the outside; the bottom stirring inner magnetic rotor 9 is driven by the bottom stirring outer magnetic rotor 11 through magnetic coupling; the bottom stirring The paddle 8 is directly connected to the inner magnetic rotor 9 for stirring at the bottom, and the inner magnetic rotor 9 for stirring at the bottom drives it to rotate to stir the liquid at the bottom of the tank body 5 .

It is worth mentioning that the present invention is a new type of magnetic stirring system for animal cell suspension culture based on the top magnetic stirring and the bottom magnetic stirring as an auxiliary. The top stirring system does not directly contact with the inside of the reactor, but passes The connection avoids the influence of the magnetic field on the inside of the reactor. The bottom magnetic stirring system is auxiliary stirring, the action time is short, and does not require a large power, so the influence of the magnetic field on the interior of the reactor can be ignored.

On the other hand, the top stirring paddle 7 and the bottom stirring paddle 8 are combined to stir, and each can achieve a good stirring effect without a high rotational speed, which effectively alleviates the radial runout problem of the stirring shaft and greatly reduces the stress on the cells. The shear force improves the mixing uniformity of the culture solution in the reactor. In addition, single-top stirring is mostly used in large-scale animal cell reactors. Top stirring cannot fully stir the liquid in the fermenter, which is prone to sedimentation. Double stirring is based on top stirring and bottom stirring system to assist stirring. this problem. The top stirring impeller 7 is the main stirring impeller, and its structure is shown in Figure 4. The top stirring impeller 7 adopts four wide-bladed stirring impellers as the axial flow type agitating impeller, which has a spiral conical curved surface blade, has a large turbulent diffusion capacity, and has a low High shear force and low power consumption, especially suitable for animal cell suspension culture process requiring mass transfer, heat transfer, solid suspension and low shear force. The bottom stirring paddle 8 is an auxiliary stirring paddle, and its structure is shown in Figure 5. The bottom stirring paddle 8 adopts oblique three-blade stirring paddles, which can produce both axial and radial shunt flow, and the flow pattern produced is better than that of straight paddles. Complex, with higher discharge energy than straight blades and relatively lower shear forces. When the stirring speed is high, it presents a turbulent dispersion state. Its large blade inclination and blade twist can make the agitator achieve a high flow field in transitional flow or even turbulent flow, and the stirring effect on the bottom sediment is more excellent.

Fig. 2 is a schematic diagram of the magnetic transmission structure of the novel magnetic stirring system of the present invention, which is used to explain the transmission principle of magnetic stirring for animal cell suspension culture bioreactors.

The driver 13 in Fig. 2 is the power source of the magnetic stirring system, drives the outer magnetic rotor 14 of built-in magnetic ring to rotate through the shaft coupling;

The inner magnetic rotor 17 of the built-in magnetic ring in Fig. 2 is driven to rotate by the outer magnetic rotor 14 through magnetic coupling; the stirring shaft 16 is connected with the inner magnetic rotor 17, and is driven to rotate by the inner magnetic rotor 17; the spacer sleeve 15 is welded on the reactor Above, it is used to isolate the inner magnetic rotor 17 installed in the reactor from the outer magnetic rotor 14 installed outside the reactor, thereby realizing a fully sealed and pollution-free animal cell suspension culture bioreactor. It can be seen that the inner magnetic rotor 17 and the outer magnetic rotor 14 are separated by the isolation sleeve 15, and the power transmission is realized through magnetic coupling. The isolation sleeve 15 is welded on the reactor, thus avoiding the possibility that there is a gap between the mechanical seal fits and the cells are easily contaminated. Problems, the fully sealed and pollution-free inside of the reactor is realized. In addition, during maintenance, it only needs to be maintained outside the reaction, the disassembly is simple and convenient, and the maintenance cost is low.

Figure 3 is an exploded schematic view of the bottom magnetic stirring device, showing the composition of the magnetic stirring system.

Figure 3 includes that the stirring paddle 8 is installed inside the reactor tank body 5 for driving the liquid in the reactor to flow and make it evenly mixed; the spacer sleeve 10 is welded on the reactor tank body 5 for isolating the outer rotor 11 and the inner rotor 9 to achieve the full seal of the reactor, the drive motor 12 drives the outer rotor 11 to rotate, and the inner rotor 9 with the inner magnetic ring driven by the magnetic coupling effect of the outer rotor 11 drives the inner rotor 9 to rotate through the magnetic force coupling, thereby driving the stirring paddle 9 to rotate to realize the liquid at the bottom of the fermentation tank of stirring.

In addition, the principle of the top stirring system is the same as that of the bottom stirring system in Fig. 3, the difference is that the stirring paddle 7 of the top stirring system is not directly connected with the driving motor, but is installed on the stirring rod 6, and the stirring rod 6 is connected with the top stirring inner magnet. Rotor 4 is connected.

A novel magnetic stirring system for animal cell suspension culture of the present invention has a simple structure, adopts double magnetic stirring mechanisms, can fully mix the liquid in the reactor, and reduces the shearing force on the cells.

In summary, the novel animal cell suspension culture magnetic stirring system of the present invention includes 1 stirring driver, 2 stirring outer magnetic rotors, 3 stirring isolation sleeves, 4 stirring inner magnetic rotors, 5 tank bodies, and 6 stirring rods , 7 top stirring paddles, 8 bottom stirring paddles, 9 bottom stirring inner magnetic rotors, 10 bottom stirring isolation sleeves, 11 bottom stirring outer magnetic rotors, 12 bottom stirring drives. The present invention mainly considers the stirring system of the animal cell suspension culture reactor, and considers its stirring effect, scale, sanitation level and the size of the shearing force suffered by the cells. Compared with mechanical stirring, the present invention adopts magnetic stirring, which is more hygienic and simple and convenient to maintain; the double stirring mechanism has two stirring paddles to stir the culture liquid, which is more sufficient to stir the culture liquid compared with ordinary single stirring, which improves the cultivation efficiency. The mixing uniformity of the liquid is also more convenient to expand the scale of the reactor; and the speed of each stirring system of the double stirring is lower than that of the single stirring, which also reduces the shear force formed by the stirring paddle. In addition, in a large-scale reactor, the stirring blade of the single stirring system cannot go deep into the tank too low, so there will still be precipitation of cells and nutrients, so the top magnetic stirring is used as the main stirring, and the bottom magnetic stirring is the auxiliary stirring to make the culture medium The mixing is more uniform and effective to solve the problem of bottom sedimentation. The invention can be effectively applied to guide the design and manufacture of bioreactors for animal cell suspension culture, which can improve the sanitation level and mixing uniformity of the animal cell suspension culture environment, reduce the shear force suffered by animal cell suspension culture, and improve animal cell suspension culture. Cell suspension culture reactor scale.

It should be pointed out that those skilled in the art can make some improvements and modifications without departing from the technical principle of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims (6)

1. The culture magnetic agitation system 1. a kind of novel zooblast suspends, it is characterised in that：Including tank body (5), and setting Top stirring system above tank body (5) and the bottom agitation system being set to below tank body (5) stir based on the top stirring system Mix system, stirring system supplemented by the bottom agitation system, the top stirring system includes power drive unit and by power drive The top agitating paddle (7) of device driving, the top agitating paddle (7) are deep into the inside of tank body (5), and the bottom agitation system includes dynamic Force driving device and the bottom agitating paddle (8) driven by power drive unit, the bottom agitating paddle (8) are set to tank body (5) bottom.
2. The culture magnetic agitation system 2. a kind of novel zooblast according to claim 1 suspends, it is characterised in that：Institute It includes driver (13), outer magnet rotor (14), interior magnet rotor (17), agitating shaft (16), separation sleeve (15) to state power drive unit, The interior magnet rotor (17) is dynamic by magnetic coupling zone of action turn by outer magnet rotor (14), agitating shaft (16) and interior magnet rotor (17) connect, driven and rotated by interior magnet rotor (17), the separation sleeve (15) is set at the top of tank body (5) or bottom, for every From interior magnet rotor (17) and outer magnet rotor (14).
3. The culture magnetic agitation system 3. a kind of novel zooblast according to claim 2 suspends, it is characterised in that：With It is to push force driving device in the power drive unit of driving top agitating paddle (7), the force driving device that pushes includes that top is stirred Driver (1), top stir outer magnet rotor (2), separation sleeve (3) is stirred on top, magnet rotor (4), stirring rod (6), top are stirred in the stirring of top Paddle (7) is mixed, the stirring rod (6) is for connecting magnet rotor (4) and top agitating paddle (7) in the stirring of top, deeply top agitating paddle (7) The liquid in tank body (5) is stirred to tank body (5).
4. The culture magnetic agitation system 4. a kind of novel zooblast according to claim 2 suspends, it is characterised in that：With It is bottom power drive unit in the power drive unit of driving bottom agitating paddle (8), the bottom power drive unit includes bottom stirring Outer magnet rotor (11), bottom stirring driver (12) are stirred in magnet rotor (9), bottom stirring separation sleeve (10), bottom in paddle (8), bottom stirring, The bottom agitating paddle (8) directly stirs interior magnet rotor (9) bottom of with and is connected, and stirring interior magnet rotor (9) bottom of by drives its rotation, stirs The liquid of tank body (5) bottom.
5. The culture magnetic agitation system 5. a kind of novel zooblast according to claim 1 suspends, it is characterised in that：Institute It is main agitating paddle to state top agitating paddle (7), uses four wide leaf agitating paddles for axial flow type agitating paddle, has screw conic curve blade.
6. The culture magnetic agitation system 6. a kind of novel zooblast according to claim 1 suspends, it is characterised in that：Institute It is auxiliary agitating paddle to state bottom agitating paddle (8), using oblique three leaves stirrer paddle.