A disposable bioreactor using a cylindrical coupled overhead magnetic stirrer

Abstract

The present invention discloses a disposable bioreactor using a cylindrical coupled top-mounted magnetic stirrer. It includes a disposable bioreactor bag, a protective shell, an inner rotor, an outer rotor, an isolation sleeve, a stirring drive motor, a stirring shaft and a stirring blade; the isolation sleeve is sleeved on the outside of the inner rotor; the outer rotor is sleeved on the outside of the isolation sleeve; an inner magnetic steel is arranged inside the inner rotor; an outer magnetic steel is arranged inside the outer rotor; the outer rotor and the inner rotor are separated by the isolation sleeve; the bottom of the isolation sleeve is bonded and fixed to the top of the bioreactor bag to seal the bag opening; the upper part of the stirring shaft passes through the center of the inner rotor and is installed in the center position of the isolation sleeve through the upper end bearing and the lower end bearing; the lower part of the stirring shaft is provided with a stirring blade; the motor output shaft of the stirring drive motor is connected to the top of the outer rotor. The disposable bioreactor has a stronger stirring torque, the bottom of the bag does not need to be supported, the stirring system does not need a bottom support, the height of the stirring blade can be adjusted according to the amount of culture medium, and the scaled-up culture in the same reactor can be realized.

Description

Technical Field

[0001] The invention belongs to the technical field of biological reaction equipment and relates to a disposable bioreactor adopting a cylindrical coupling overhead magnetic stirrer. Background Art

[0002] With the development of biological cell culture technology, stirred bioreactors are increasingly used in biological cell culture. Traditional stirred bioreactors use stainless steel bioreactors; the existing overhead magnetic stirrer for stainless steel bioreactors includes: a shell, a driving device and a driving shaft arranged on the shell, a driving rotor arranged in the shell and connected to the driving shaft, and a driven rotor arranged corresponding to the driving rotor; a magnetic steel is arranged inside the driving rotor and the driven rotor; a metal isolation cover is arranged between the driving rotor and the driven rotor; the stirring shaft is connected to the driven rotor; a bearing seat supports and connects the stirring shaft; a splitter cover is arranged between the stirring shaft and the bearing seat; there is a first channel between the splitter cover and the bearing seat and the stirring shaft; and there is a second channel between the bearing seat and the isolation cover at the bottom of the bearing seat. In other words, in the existing overhead magnetic stirrer for stainless steel bioreactors, the stirring shaft needs to be supported by a bearing seat, and the magnetic stirrer is connected to the stainless steel bioreactor through a metal bearing seat (flange bolt connection).

[0003] Disposable bioreactors (SUBs) are specifically used to replace traditional stainless steel bioreactors (stirred biofermentation tanks) for animal cell culture. Like traditional stainless steel bioreactors, disposable bioreactors are also used to culture cells or microorganisms (mainly for pharmaceutical purposes). Unlike traditional stainless steel bioreactors, disposable bioreactors (also called bag reactors) contain culture fluid and culture medium in a disposable plastic bag (i.e., disposable bioreactor bag) made of special plastic with sufficient thickness and toughness. This plastic bag (disposable bioreactor bag) needs to be placed in a stainless steel protective shell (i.e., steel shell) for use.

[0004] Disposable bioreactor bags (BPC bags) are sterilized by radiation and can be used in various pharmaceutical processes: such as preparation, buffering, temporary storage, mixing, and liquid preparation, especially for the culture of cells and bacteria. Disposable bioreactor bags are generally discarded after only one use. They do not require a cleaning system or a sterilization system, which avoids cross contamination, saves a lot of time and space, and greatly reduces costs. A stirrer must be installed on the disposable bioreactor bag to ensure sufficient stirring during the cell culture process and ensure the healthy growth of cells.

[0005] There are three types of stirring methods for existing disposable bioreactor bags: a. bottom-mounted magnetic stirring; b. top-mounted mechanical stirring; c. end-face coupled top-mounted magnetic stirring. Bottom-mounted magnetic stirring cannot provide satisfactory mixing performance in larger bags. Top-mounted mechanical stirring has a greater risk of use due to leakage.

[0006] The existing end-coupled overhead magnetic stirrer for bioreactor bags uses Figure 1 The end-face coupling structure shown (the outer rotor 2 and the inner rotor 1 are arranged horizontally up and down, the outer magnetic steel 4 and the inner magnetic steel 3 are arranged opposite to each other up and down, and the inner rotor 1 and the outer rotor 2 are separated by the isolation plate 50) must be provided with a fulcrum (bottom bearing 31 and bottom support 32) at the bottom of the bag to stabilize the stirring shaft. This end-face coupling top-mounted magnetic stirring structure has the following disadvantages:

[0007] 1. Once the culture solution and culture medium are poured into the bag, the up and down position of the stirring blade cannot be adjusted, so it is not convenient to adjust the height of the stirring blade according to the amount of culture solution and culture medium.

[0008] Cell culture is the process of gradually culturing a single cell or a small number of cells into a large number of cells. When using a bioreactor bag for cell culture, the cell culture fluid in the bioreactor bag may be 10L at the beginning. After a period of culture, the culture medium is added to 50L. After a period of continued culture, the culture medium is added to 250L. After a period of continued culture, the culture medium is added to 1000L, and the culture is continued for a period of time. Therefore, the height of the stirring blade needs to be adjusted according to the amount of culture fluid and culture medium in the bioreactor bag.

[0009] 2. The fulcrum at the bottom of the bag, because of the bearing, is not only prone to wear debris, but also easy to get stuck (when there are solid particles in the culture medium).

[0010] 3. As a biological culture equipment that meets the requirements of GMP (Good Manufacturing Practice for Pharmaceuticals), its design principle is that the internal structure should be simple, so it is a better design to eliminate the bottom support of the bag. Summary of the invention

[0011] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a disposable bioreactor using a cylindrical coupled overhead magnetic stirrer that does not require a support point at the bottom of the bag and a stirring system that does not require a bottom support, and can adjust the height of the stirring blade according to the amount of culture medium, so as to achieve scaled culture in the same reactor.

[0012] The objective of the present invention is achieved through the following technical solutions:

[0013] The present invention discloses a disposable bioreactor using a cylindrical coupled overhead magnetic stirrer, comprising a disposable bioreactor bag, a protective shell (steel shell) arranged outside the disposable bioreactor bag, and a cylindrical coupled overhead magnetic stirrer; the cylindrical coupled overhead magnetic stirrer comprises an inner rotor, an outer rotor, an inner magnetic steel, an outer magnetic steel, an isolation sleeve, a stirring drive motor, a stirring shaft, a stirring blade, an upper end bearing and a lower end bearing; the inner rotor is a cylinder; the isolation sleeve is sleeved outside the inner rotor; the outer rotor is sleeved outside the isolation sleeve; the inner rotor is provided with an inner magnetic steel; the outer rotor is provided with a The invention discloses a method for preparing a stirring shaft comprising: providing an outer magnetic steel; the inner magnetic steel on the inner rotor and the outer magnetic steel on the outer rotor are arranged vertically inside and outside opposite to each other; the outer rotor and the inner rotor are separated by an isolation sleeve; the upper end bearing and the lower end bearing are fixedly installed on the inner wall of the isolation sleeve; the upper part of the stirring shaft passes through the center of the inner rotor, and is installed at the center position of the isolation sleeve through the upper end bearing and the lower end bearing; the lower part of the stirring shaft is provided with a stirring blade; the motor output shaft of the stirring drive motor is connected with the top end of the outer rotor; the isolation sleeve is made of engineering plastic, and the bottom of the isolation sleeve can be bonded and fixed to the top of the disposable bioreactor bag, and can seal the top bag opening of the disposable bioreactor bag.

[0014] Furthermore, an upright lifting rod is provided beside the protective shell (steel shell) outside the disposable bioreactor bag, and upper and lower cross arms are respectively installed on the upper and middle parts of the lifting rod, the upper cross arm is a motor fixing beam, and the lower cross arm is a bag clamp; the stirring drive motor is installed and fixed on the motor fixing beam; the front end of the bag clamp is provided with a fixing clamp that can clamp the outer part of the lower part of the isolation sleeve on the disposable bioreactor bag.

[0015] Furthermore, a plurality of slopes, namely guide slopes, are arranged from top to bottom on the outer wall of the isolation sleeve.

[0016] Furthermore, upper rollers or balls and lower rollers or balls are embedded on the inner wall of the outer rotor, and are respectively arranged above and below the outer magnetic steel; there are multiple upper rollers or balls evenly distributed along the circumference, and there are multiple lower rollers or balls evenly distributed along the circumference.

[0017] Furthermore, a central opening is provided on the bottom plate of the isolation sleeve; a protective plate is provided at the connection between the central opening of the bottom plate of the isolation sleeve and the stirring shaft; the upper part of the protective plate is an annular sleeve with a central hole; the lower part of the protective plate is a downwardly protruding conical sleeve with a central hole; the annular sleeve on the upper part of the protective plate is embedded in the central opening of the bottom plate of the isolation sleeve; the stirring shaft passes through the central opening of the protective plate; the annular sleeve and the conical sleeve of the protective plate are sleeved outside the stirring shaft.

[0018] Furthermore, the motor output shaft of the stirring drive motor is connected to the top end of the outer rotor through a flexible connector (aluminum alloy or polyamide glass fiber reinforced material).

[0019] Furthermore, the upper end bearing and the lower end bearing are respectively arranged above and below the inner rotor, or the upper end bearing and the lower end bearing are both arranged below the inner rotor.

[0020] Furthermore, the fixing fixture is composed of a clamping hand with a semicircular groove and a clamping block with a semicircular groove connected (joined) by screws.

[0021] Furthermore, the lifting rod is provided with a driving part lifting motor connected with the motor fixing beam and capable of pushing the motor fixing beam to rise. The driving part lifting motor can push the motor fixing beam to rise.

[0022] Furthermore, the lifting rod is provided with a stirring system lifting motor connected to the lifting rod and capable of pushing the lifting rod upward. The stirring system lifting motor can push the lifting rod upward to drive the upper cross arm and the lower cross arm to rise together, and at the same time drive the stirring shaft and stirring blades to rise.

[0023] Furthermore, the cylindrical coupled overhead magnetic stirrer includes multiple groups of magnetic steels; each group of magnetic steels includes an inner magnetic steel and an outer magnetic steel, and the two are arranged opposite to each other; the multiple groups of magnetic steels are evenly distributed along the circumference and symmetrically arranged; the number of inner magnetic steels on the inner rotor is the same as the number of outer magnetic steels on the outer rotor.

[0024] Furthermore, both the inner rotor and the outer rotor are blocks of non-magnetic materials; the inner rotor has four, six, eight or more grooves evenly distributed and symmetrically arranged along the circumference on the outer side, and a magnet is embedded in each groove, namely the inner magnet; the outer rotor has four, six, eight or more grooves evenly distributed and symmetrically arranged along the circumference on the inner side, and a magnet is embedded in each groove, namely the outer magnet.

[0025] Beneficial effects of the present invention:

[0026] The present invention designs a disposable bioreactor using a cylindrical coupling overhead magnetic stirrer, the main feature of which is that a cylindrical coupling overhead magnetic stirrer (such as Figure 2 As shown, the inner rotor 1 and the outer rotor 2 are arranged vertically inside and outside, the inner magnetic steel 3 and the outer magnetic steel 4 are arranged vertically inside and outside oppositely, and the inner rotor 1 and the outer rotor 2 are separated by an isolation sleeve 5), instead of the overhead magnetic stirrer using the end face coupling method in the prior art (such as Figure 1 As shown, the outer rotor 2 and the inner rotor 1 are arranged horizontally up and down, the outer magnetic steel 4 and the inner magnetic steel 3 are arranged opposite to each other up and down, and the inner rotor 1 and the outer rotor 2 are separated by an isolation plate 50). The advantages of this structure are: no support point is required at the bottom of the bag (the most desired improvement by the user), and the stirring system does not need bottom support; because the fixed support point is eliminated at the bottom of the bag, the upper and lower positions of the stirring blades can be adjusted at any time during operation as needed, thereby realizing scaled-up culture in the same reactor.

[0027] The comparison of magnetic coupling efficiency and stirring torque between the end face coupling method and the cylindrical coupling method is as follows:

[0028] End coupling method: Figure 1 As shown, in order to leave a gap between the inner rotor and the isolation plate, a fulcrum is required to balance the attraction (magnetic force) between the inner and outer rotors. If a larger torque is required, there are two ways to increase the torque: 1. Increase the area of ​​the magnetic steel; 2. Reduce the gap between the inner and outer rotors. Both methods will increase the attraction between the inner and outer rotors. This attraction needs to be offset by the support point 40 (usually a bearing). Therefore, the support point 40 needs to be strong enough. The greater the attraction, the greater the friction. The friction requires loss torque, which reduces the magnetic coupling efficiency. The greater the friction, the greater the loss torque, which reduces the stirring torque and reduces the magnetic coupling efficiency. On the other hand, the hard plastic isolation plate 50 on the top of the bag has a limited area. If its area is increased, the thickness of the isolation plate 50 needs to be increased, which in turn reduces the magnetic coupling efficiency.

[0029] Cylindrical coupling method: Figure 2 , Figure 6 As shown, the attraction (magnetic force) between the inner and outer rotors is balanced on the stirring shaft (as shown in Figure 2 As shown in the direction of the two arrows, on the same cross section, the two magnetic forces F1 and F2 acting on the stirring shaft are in opposite directions, one to the left and the other to the right, which cancel each other out. The radial load on the bearing is relatively light, or very light (if the processing accuracy is good). Therefore, the friction of the bearing will be relatively small. With small friction, the torque loss is small, which increases the stirring torque and improves the magnetic coupling efficiency. If a larger torque is required, there are two ways to increase the torque: 1. Increase the length of the inner and outer rotors (that is, increase the coupling area); 2. Or appropriately increase the diameter of the inner and outer rotors (also increase the coupling area). There is a lot of room to increase the length of the inner and outer rotors, and there is basically no need to increase the thickness of the isolation sleeve 5.

[0030] Compared with the prior art, the disposable bioreactor using a cylindrical coupled overhead magnetic stirrer of the present invention has the following advantages:

[0031] 1. The bottom of the bag does not require a support point, and the mixing system does not require a bottom support.

[0032] 2. There is no support point at the bottom of the bag, which is more in line with GMP requirements and reduces the failure rate.

[0033] 3. Once the bag is filled with culture medium, the position of the stirring blade can still be adjusted appropriately by moving the entire magnetic stirrer down or up.

[0034] 4. Because the position of the stirring blade can be adjusted appropriately, the mixing performance can be optimized at different stages of culture to improve cell activity.

[0035] 5. Because the position of the stirring blade can be adjusted appropriately, the cell culture can be scaled up in one bag, and the scaled up culture can be achieved in the same reactor.

[0036] 6. The cylindrical coupling method is adopted, the magnetic coupling efficiency is higher, and it can provide stronger stirring torque.

[0037] 7. The sealing and isolation performance is better, and the mixing system will not produce any impurities or debris that endanger the product quality, which is more suitable for the terminal liquid distribution system with high cleanliness requirements.

[0038] 8. Easy to install and operate. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 It is a simplified structural schematic diagram of an overhead magnetic stirrer using an end-face coupling method in the prior art;

[0040] Figure 2 It is a simplified structural schematic diagram of an overhead magnetic stirrer using a cylindrical coupling method in the present invention;

[0041] Figure 3 It is a schematic diagram of the structure of a disposable bioreactor bag using an end-face coupled overhead magnetic stirrer in the prior art;

[0042] Figure 4 It is a schematic structural diagram of a disposable bioreactor using a cylindrical coupled overhead magnetic stirrer of the present invention;

[0043] Figure 5 It is a structural schematic diagram of the steel shell 19, the lifting rod 15, and the upper and lower cross arms of the disposable bioreactor bag in the present invention;

[0044] Figure 6 It is a detailed structural schematic diagram of the overhead magnetic stirrer of the cylindrical coupling mode in the present invention;

[0045] Figure 7 It is a schematic diagram of the structure of the outer rotor 2 in the present invention;

[0046] Figure 8 It is a schematic structural diagram of the connection between the inner rotor 1, the upper end bearing 9 and the stirring shaft 7 in the present invention;

[0047] Fig. 9 It is a schematic structural diagram of the isolation sleeve 5 in the present invention;

[0048] Fig.10 It is a schematic diagram of the structure of the protective plate 16 in the present invention;

[0049] Figure 11-13 1 is a schematic structural diagram of the lower cross arm 12 and the fixing fixture 120 in the present invention;

[0050] Fig.14 It is a schematic diagram of the structure in which the stirring drive motor 6 in the present invention is installed on the motor fixing beam 11;

[0051] Fig.15 It is a schematic diagram of the installation of the motor fixing beam 11 and the lower cross arm 12 in the present invention;

[0052] Figure 16-Figure 19 It is a schematic diagram of placing a disposable bioreactor bag 20 in a steel shell 19 in the present invention;

[0053] Figure 20-22 It is a schematic diagram of gradually sliding the outer rotor 2 and the stirring drive motor 6 assembly into the outside of the isolation sleeve in the present invention;

[0054] Figure 23-25 It is a schematic diagram showing that the position of the stirring blade 8 can be raised along with the increase of the liquid level of the culture solution 21 in the present invention.

[0055] In the figure: 1, inner rotor 2, outer rotor 200, motor connection end 3, inner magnetic steel 4, outer magnetic steel 5, isolation sleeve 51, guide slope 52, guide slope 53, positioning block 54, clamping fixing part 55, center opening 6, stirring drive motor 7, stirring shaft 8, stirring blade 9, upper end bearing 10, lower end bearing 11, motor fixing beam 12, lower cross arm 120, fixing fixture 121, clamping hand 122, clamping block 123, screw 13, driving part lifting motor 14, stirring system lifting motor 15, lifting rod 16, protective plate 160, center hole 161, annular sleeve 162, conical sleeve 17, upper roller 18, lower roller 19, steel shell 20, disposable bioreactor bag 21, culture medium 31, bottom bearing 32, bottom support 40, support point 50, isolation plate DETAILED DESCRIPTION

[0056] The present invention is further described below in conjunction with the accompanying drawings and embodiments. Example

[0057] like Figure 4 , Figure 5As shown, the present invention is a disposable bioreactor using a cylindrical coupled overhead magnetic stirrer, comprising a disposable bioreactor bag 20, a protective shell (steel shell 19) arranged outside the disposable bioreactor bag 20, and a cylindrical coupled overhead magnetic stirrer; the cylindrical coupled overhead magnetic stirrer comprises an inner rotor 1, an outer rotor 2, an inner magnetic steel 3, an outer magnetic steel 4, an isolation sleeve 5, a stirring drive motor 6, a stirring shaft 7, a stirring blade 8, an upper end bearing 9, and a lower end bearing 10; the inner rotor 1 is a cylinder, the outer rotor 2 is an inverted U-shaped cover body, and the isolation sleeve 5 is an inverted U-shaped cover; the isolation sleeve 5 is sleeved on the outside of the inner rotor 1; the outer rotor 2 is sleeved on the upper and middle outer parts of the isolation sleeve 5. surface; the inner rotor 1 is arranged in the upper middle position of the isolation sleeve 5; the inner rotor 1 is embedded with an inner magnetic steel 3 on the outer side, and the inner magnetic steel 3 is evenly distributed and symmetrically arranged in multiple pieces (four pieces, six pieces or eight pieces) along the circumference; the outer magnetic steel 4 is embedded with an outer magnetic steel 4 on the inner rotor 2 on the inner side, and the outer magnetic steel 4 is evenly distributed and symmetrically arranged in multiple pieces (four pieces, six pieces or eight pieces) along the circumference; the inner magnetic steel 3 on the inner rotor 1 and the outer magnetic steel 4 on the outer rotor 2 are arranged vertically inside and outside oppositely; there is an isolation sleeve 5 between the outer magnetic steel 4 and the inner magnetic steel 3, and they are isolated by the isolation sleeve 5; the isolation sleeve 5 is made of hard engineering plastic material, and the bottom of the isolation sleeve 5 is bonded and fixed to the top of the disposable bioreactor bag 20, and can seal the top bag opening of the disposable bioreactor bag 20.

[0058] like Figure 4 , Figure 6 , Figure 8 As shown, the upper bearing 9 and the lower bearing 10 are respectively arranged above and below the inner rotor 1, and are fixedly mounted on the inner wall of the inverted U-shaped isolation sleeve 5; the upper part of the stirring shaft 7 (with a reduced diameter) passes through the central hole of the inner rotor 1, and is installed at the upper center position in the isolation sleeve 5 through the upper bearing 9; the middle part of the stirring shaft 7 is installed at the middle center position in the isolation sleeve 5 through the lower bearing 10; and a plurality of stirring blades 8 are provided at the lower part of the stirring shaft 7.

[0059] like Figure 4 , Figure 6 As shown, the cylindrical coupled overhead magnetic stirrer includes multiple groups (four groups, six groups or eight groups) of magnetic steels; each group of magnetic steels includes an inner magnetic steel and an outer magnetic steel, and two pieces are arranged opposite to each other in each group; multiple groups of magnetic steels are evenly distributed along the circumference and symmetrically arranged. The number of inner magnetic steels 3 on the inner rotor 1 is the same as the number of outer magnetic steels 4 on the outer rotor 2; each magnetic steel (including the inner magnetic steel 3 and the outer magnetic steel 4) is the same size (that is, the length, height and thickness are the same). The inner rotor 1 and the outer rotor 2 are both stainless steel blocks; the inner rotor 1 has four, six or eight grooves dug on the outer side of the stainless steel block, and each groove is embedded with a magnetic steel, namely the inner magnetic steel 3; the outer rotor 2 has four, six or eight grooves dug on the inner side of the stainless steel block, and each groove is embedded with a magnetic steel, namely the outer magnetic steel 4.

[0060] like Figure 4 , Figure 5, Fig.15 As shown, the disposable bioreactor bag 20 needs to be placed in a stainless steel protective shell (steel shell 19) when in use. A vertical lifting rod 15 is provided beside the stainless steel protective shell (steel shell 19) outside the disposable bioreactor bag 20. An upper and lower cross arms are respectively installed on the upper and middle parts of the lifting rod 15. The upper cross arm (steel folding plate) is the motor fixing beam 11, and the lower cross arm 12 (square steel pipe) is the bag clamp; the stirring drive motor 6 is installed and fixed on the motor fixing beam 11. Figure 11-13 As shown, the bag holder, i.e., the lower cross arm 12, is provided at the front end with a clamping hand 121 with a semicircular groove and a clamping block 122 with a semicircular groove connected and fixed by screws 123, which can clamp the fixing fixture 120 outside the clamping fixing portion 54 at the bottom of the isolation sleeve 5, and can clamp and fix the disposable bioreactor bag 20. Fig.14 As shown, a servo motor flange is provided on the motor fixing beam 11, and the stirring drive motor 6 is fixed on the motor fixing beam 11 through the servo motor flange. The motor output shaft of the stirring drive motor 6 is connected to the top protruding end of the outer rotor 2 (connecting to the motor end 200) through a flexible connector (aluminum alloy or polyamide glass fiber reinforced material).

[0061] like Figure 5 As shown, the lifting rod 15 is provided with a driving part lifting motor 13 connected to the upper cross arm (motor fixed beam 11), and is also provided with a stirring system lifting motor 14 connected to the lifting rod 15; the driving part lifting motor 13 can push the motor fixed beam 11 to rise; the stirring system lifting motor 14 can push the lifting rod 15 to rise, driving the upper cross arm (motor fixed beam 11) and the lower cross arm 12 to rise together, thereby driving the stirring shaft 7 and the stirring blades 8 to rise.

[0062] like Figure 6 , Figure 7 As shown, the inner wall of the inverted U-shaped outer rotor 2 is embedded with upper rollers 17 and lower rollers 18, which are respectively arranged above and below the outer magnetic steel 4. There are multiple upper rollers 17 evenly distributed along the circumference, and there are multiple lower rollers 18 evenly distributed along the circumference. The purpose of this design is to facilitate the outer rotor 2 to slide smoothly into the outer side of the isolation sleeve 5 when it is installed outside the isolation sleeve 5.

[0063] like Figure 4 , Fig. 9 As shown, a slope, namely a guide slope 51, 52, is provided on the outer wall of the upper and middle parts of the isolation sleeve 5 (there is a guide slope along the circumference, one circle at the top and one circle at the bottom). The purpose of this design is also to facilitate the outer rotor 2 to slide smoothly into the outside of the isolation sleeve 5 when it is installed on the outside of the isolation sleeve 5. A circle of protruding positioning blocks 53 is provided on the outer wall of the lower part of the isolation sleeve 5. Below the positioning blocks 53 at the lower part of the isolation sleeve 5 is a clamping fixing part 54; the bottom plate of the isolation sleeve 5 is provided with a central opening 55.

[0064] like Figure 4 , Fig.10 As shown, a protective plate 16 is additionally provided at the connection between the central opening 55 of the bottom plate of the isolation sleeve 5 and the stirring shaft 7; the upper part of the protective plate 16 is an annular sleeve 161 with a central hole 160; the lower part of the protective plate 16 is a downwardly protruding conical sleeve 162 with a central hole 160; the annular sleeve 161 at the upper part of the protective plate 16 is embedded in the central opening 55 of the bottom plate of the isolation sleeve 5; the stirring shaft 7 passes through the central hole 160 of the protective plate 16; the annular sleeve 161 and the conical sleeve 161 of the protective plate 16 are sleeved on the outside of the stirring shaft 7; there is only a 0.1 mm gap between the isolation sleeve 5 and the protective plate 16 and the stirring shaft 7, which can reduce the escape of culture solution, foam and / or water vapor.

[0065] A liquid injection port is provided on the side of the disposable bioreactor bag 20, through which a culture fluid 21 (liquid culture medium inoculated with cells) can be injected for cell culture.

[0066] The disposable bioreactor of the present invention using a cylindrical coupled overhead magnetic stirrer has the following innovative designs:

[0067] 1) Since the soft bag (disposable bioreactor bag 20, disposable, used for cell and bacteria culture or liquid preparation, mixing, etc.) is discarded after use, the operation of installing the soft bag into the steel shell 19 and connecting it to the outer rotor 2 is a frequent task in daily life. Therefore, how to quickly and easily install and fix the soft bag and connect it to the motor drive end is a problem that needs to be considered and solved.

[0068] The design of the present invention eliminates the need for the outer rotor of the existing magnetic agitator to be fixed by a steel structure of the motor base, and instead directly sleeves it on the isolation sleeve of the soft bag, and adopts rollers or balls (upper roller 17, lower roller 18) arranged up and down to fit and connect with the isolation sleeve 5, thereby fixing the outer rotor 2 and ensuring the precision requirement of the concentricity of the outer rotor 2 with the isolation sleeve 5 during the movement.

[0069] 2) In order to enable the magnetic stirrer driving part (outer rotor 2 + stirring driving motor 6) to be easily separated from or fitted with the isolation sleeve 5, an upper cross arm (motor fixing beam 11) which can move up and down and automatically position is provided.

[0070] 3) The isolation sleeve 5 is designed to have two guide slopes (guide slopes 51, 52), so that the outer rotor 2 can slide in smoothly and be accurately positioned.

[0071] 4) In order to ensure that the soft bag can be firmly fixed, a "neck" that can be clamped is provided on the isolation sleeve 5 (ie, the clamping and fixing portion 54 of the isolation sleeve 5).

[0072] 5) In order to enable the stirring blade 8 to move up and down, the entire stirrer can be driven by the stirring system lifting motor 14 to move up and down.

[0073] 6) In order to prevent the culture solution, foam and / or water vapor from entering the isolation sleeve 5 of the magnetic stirrer, a protective plate 16 is provided.

[0074] 7) All materials should be sterilized by irradiation.

[0075] The working principle of the cylindrical coupled overhead magnetic stirrer in the present invention is as follows: the stirring drive motor 6 is started, driving the outer rotor 2 to rotate; there is a magnetic field between the outer rotor 2 and the inner rotor 1 (between the outer magnetic steel 4 and the inner magnetic steel 3); through the action of the magnetic field, the outer rotor 2 (driving rotor) transmits the torque to the inner rotor 1 (driven rotor), thereby driving the inner rotor 1 to rotate, and the rotation of the inner rotor 1 in turn drives the stirring shaft 7 and the stirring blade 8 to rotate, thereby achieving stirring of the culture solution 21 (liquid culture medium inoculated with cells) in the disposable bioreactor bag 20.

[0076] The installation and use method of the disposable bioreactor using a cylindrical coupled overhead magnetic stirrer of the present invention is as follows:

[0077] First, the upper end bearing 9 is embedded in the inner wall of the inverted U-shaped isolation sleeve 5; then the upper part of the stirring shaft 7 passes through the central hole of the inner rotor 1, and the upper end of the stirring shaft 7 is installed in the upper end bearing 9, and the inner rotor 1 is placed in the center of the isolation sleeve 5; then, the lower end bearing 10 is embedded in the inner wall of the inverted U-shaped isolation sleeve 5, and the upper part of the stirring shaft 7 is installed in the lower end bearing 10; then, the middle and lower part of the stirring shaft 7 is inserted into the disposable bioreactor bag 20 from the bag opening at the top, and then the bottom of the isolation sleeve 5 is bonded and fixed to the top of the disposable bioreactor bag 20 to seal the top bag opening of the disposable bioreactor bag 20.

[0078] Upper and lower cross arms are respectively installed on the upper and middle parts of the lifting rod 15 beside the steel shell 19 of the disposable bioreactor; the distance between the lower cross arm 12 and the upper cross arm, i.e., the motor fixing beam 11, can be appropriately adjusted.

[0079] The disposable bioreactor bag 20 equipped with the stirring shaft 7, the inner rotor 1 and the isolation sleeve 5 is placed in the steel shell 19; the fixing fixture 120 at the front end of the lower cross arm 12 is used to clamp the outer side of the clamping and fixing portion 54 at the lower part of the isolation sleeve 5 (first remove the clamping block 122 with the semicircular groove, put half of the clamping and fixing portion 54 at the lower part of the isolation sleeve 5 into the semicircular groove of the clamping hand 121, and then put the semicircular groove of the clamping block 122 on the outer side of the other half of the clamping and fixing portion 54 at the lower part of the isolation sleeve 5, and finally close the clamping block 122 and the clamping hand 121 and fix them with screws), so that the disposable bioreactor bag 20 can be clamped and fixed.

[0080] The stirring drive motor 6 is fixed on the upper cross arm, i.e., the motor fixing beam 11. During installation, it should be noted that the motor shaft center of the stirring drive motor 6 is on the same vertical line as the center of the isolation sleeve 5. The motor shaft of the stirring drive motor 6 is connected to the top protruding end of the outer rotor 2 (with the motor connection end 200) by a flexible connector (aluminum alloy or polyamide glass fiber reinforced material).

[0081] like Figure 16-Figure 19 As shown, the upper cross arm, i.e., the motor fixing beam 11, is moved downward, and the outer rotor 2 and the stirring drive motor 6 assembly are installed outside the isolation sleeve 5: Figure 20-22 As shown, the first is automatic alignment (such as Fig. 20 ), then gradually slide in (as shown Fig.21 As shown) is placed outside the isolation sleeve 5, and finally positioned (as shown Fig. 22 As shown in the figure, the outer magnetic steel 4 on the outer rotor 2 and the inner magnetic steel 3 on the inner rotor 1 have the same height and position, and are opposite to each other.

[0082] After the cylindrical coupling top-mounted magnetic stirrer is installed on the disposable bioreactor bag 20, it can be used for cell culture. When in use, the culture solution 21 (liquid culture medium inoculated with cells) is injected from the injection port on the side of the disposable bioreactor bag 20 for cell culture. During the cell culture process, the stirring drive motor 6 is turned on to drive the outer rotor 2 to rotate, and the outer rotor 2 drives the inner rotor 1 to rotate. The rotation of the inner rotor 1 drives the stirring shaft 7 and the stirring blade 8 to rotate, and the culture solution 21 (liquid culture medium inoculated with cells) in the disposable bioreactor bag 20 is stirred.

[0083] In the disposable bioreactor bag using the end-face coupled top-mounted magnetic stirrer of the present invention, the upper and lower positions of the stirring blades 8 can be adjusted at any time during operation as needed. During installation, the installation position of the stirring blades 8 can be appropriately adjusted, and the distance between the lower cross arm 12 and the upper cross arm, i.e., the motor fixing beam 11, can be appropriately adjusted by driving the lifting motor 13 of the driving part; once the culture medium is infused into the bag, the position of the stirring blades 8 can still be appropriately adjusted - just start the stirring system lifting motor 14 to move the entire magnetic stirrer downward or upward. Figure 23-25 As shown, as the level of the culture solution 21 increases, the position of the stirring blade 8 can also be increased accordingly.

Claims

1. A disposable bioreactor using a cylindrical coupled overhead magnetic stirrer, characterized in that: The invention comprises a disposable bioreactor bag, a protective shell arranged outside the disposable bioreactor bag, and a cylindrical coupling top-mounted magnetic stirrer; the cylindrical coupling top-mounted magnetic stirrer comprises an inner rotor, an outer rotor, an inner magnetic steel, an outer magnetic steel, an isolation sleeve, a stirring drive motor, a stirring shaft, a stirring blade, an upper end bearing and a lower end bearing; the inner rotor is a cylinder; the isolation sleeve is sleeved outside the inner rotor; the outer rotor is sleeved outside the isolation sleeve; an inner magnetic steel is arranged inside the inner rotor; an outer magnetic steel is arranged inside the outer rotor; the inner magnetic steel on the inner rotor and the outer magnetic steel on the outer rotor are arranged vertically inside and outside oppositely; the outer rotor and the inner rotor are isolated by the isolation sleeve; the upper end bearing and the lower end bearing are fixedly installed on the inner wall of the isolation sleeve; the upper part of the stirring shaft passes through the center of the inner rotor and is installed in the center position of the isolation sleeve through the upper end bearing and the lower end bearing; the lower part of the stirring shaft is provided with a stirring blade; the motor output shaft of the stirring drive motor is connected to the top end of the outer rotor; the isolation sleeve is made of engineering plastic material, and the bottom of the isolation sleeve can be bonded and fixed to the top of the disposable bioreactor bag to seal the top bag opening of the disposable bioreactor bag; A vertical lifting rod is provided beside the protective shell outside the disposable bioreactor bag, and upper and lower cross arms are respectively installed on the upper and middle parts of the lifting rod, the upper cross arm is a motor fixing beam, and the lower cross arm is a bag clamp; the stirring drive motor is installed and fixed on the motor fixing beam; the front end of the bag clamp is provided with a fixing clamp that can clamp the outer side of the lower part of the isolation sleeve on the disposable bioreactor bag; the lifting rod is provided with a stirring system lifting motor connected to the lifting rod and capable of pushing the lifting rod to rise; the lifting rod is provided with a driving part lifting motor connected to the motor fixing beam and capable of pushing the motor fixing beam to rise; The inner wall of the outer rotor is embedded with an upper roller or ball and a lower roller or ball, which are respectively arranged above and below the outer magnetic steel; a plurality of upper rollers or balls are evenly distributed along the circumference, and a plurality of lower rollers or balls are evenly distributed along the circumference; The bag clamp, i.e., the front end of the lower cross arm, is provided with a clamping hand with a semicircular groove and a clamping block with a semicircular groove, which are connected and fixed by screws; a circle of protruding positioning blocks is provided on the outer wall of the lower part of the isolation sleeve, and the clamping and fixing part is below the positioning block at the lower part of the isolation sleeve; the clamping block with the semicircular groove is taken down, and half of the clamping and fixing part at the lower part of the isolation sleeve is placed into the semicircular groove of the clamping hand, and then the semicircular groove of the clamping block is put on the outside of the other half of the clamping and fixing part at the lower part of the isolation sleeve, and finally the clamping block and the clamp are closed and fixed with screws.

2. The disposable bioreactor using a cylindrical coupled overhead magnetic stirrer according to claim 1, characterized in that: The protective shell is a steel shell.

3. The disposable bioreactor using a cylindrical coupled overhead magnetic stirrer according to claim 1, characterized in that: A plurality of slopes, namely guide slopes, are arranged from top to bottom on the outer wall of the isolation sleeve.

4. The disposable bioreactor using a cylindrical coupled overhead magnetic stirrer according to claim 1, characterized in that: A central opening is provided on the bottom plate of the isolation sleeve; a protective plate is provided at the connection between the central opening of the bottom plate of the isolation sleeve and the stirring shaft; the upper part of the protective plate is an annular sleeve with a central hole; the lower part of the protective plate is a downwardly protruding conical sleeve with a central hole; the annular sleeve on the upper part of the protective plate is embedded in the central opening of the bottom plate of the isolation sleeve; the stirring shaft passes through the central opening of the protective plate; the annular sleeve and the conical sleeve of the protective plate are sleeved outside the stirring shaft.

5. The disposable bioreactor using a cylindrical coupled overhead magnetic stirrer as claimed in claim 1, characterized in that ,The motor output shaft of the stirring drive motor is connected to the top of the outer rotor through a flexible connector.

6. The disposable bioreactor using a cylindrical coupled overhead magnetic stirrer according to claim 1, characterized in that: The upper end bearing and the lower end bearing are respectively arranged above and below the inner rotor, or the upper end bearing and the lower end bearing are both arranged below the inner rotor.

7. The disposable bioreactor using a cylindrical coupled overhead magnetic stirrer according to claim 1, characterized in that: The cylindrical coupling overhead magnetic stirrer for disposable bioreactor bags comprises a plurality of sets of magnetic steels; each set of magnetic steels comprises an inner magnetic steel and an outer magnetic steel, the two magnetic steels being arranged opposite to each other; The plurality of groups of magnetic steels are evenly distributed along the circumference and symmetrically arranged; the number of the inner magnetic steels on the inner rotor is the same as the number of the outer magnetic steels on the outer rotor.

8. The disposable bioreactor using a cylindrical coupled overhead magnetic stirrer according to claim 1, characterized in that: Both the inner rotor and the outer rotor are blocks of non-magnetic materials; the inner rotor has four, six, or eight grooves evenly distributed and symmetrically arranged along the circumference on the outer side, and each groove is embedded with a magnet, namely the inner magnet; the outer rotor has four, six, or eight grooves evenly distributed and symmetrically arranged along the circumference on the inner side, and each groove is embedded with a magnet, namely the outer magnet.

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