A disposable stirred-tank cell bioreactor
By combining the positioning short shaft clamping assembly, the cell culture bag assembly, and the stirring shaft rotating assembly, the problems of insufficient sealing and uneven stirring in the prior art are solved. This achieves fully enclosed sealing and uniform stirring of the cell culture bag, improves mass transfer efficiency and dissolved oxygen supply, and enhances cell culture results.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- 惠倪
- Filing Date
- 2023-06-29
- Publication Date
- 2026-06-26
AI Technical Summary
Existing disposable stirred tank cell bioreactors suffer from problems such as insufficient sealing, uneven mixing, low mass transfer efficiency, and insufficient dissolved oxygen, which affect cell culture results.
It adopts a combined design of positioning short shaft clamping assembly, cell culture bag assembly, stirring shaft rotating assembly and drive mechanism assembly, including V-shaped chuck, movable V-shaped chuck, irregular silicone tubing, radial magnetic coupler, etc., to achieve fully enclosed sealing and uniform stirring. The stirring shaft is driven to rotate by magnetic force, and multiple stirring paddles are installed to improve stirring efficiency.
This technology achieves a fully sealed cell culture bag, improving mixing uniformity and mass transfer efficiency, enhancing dissolved oxygen supply, reducing cell death, increasing cell density and activity, reducing the risk of contamination, and shortening the production cycle.
Smart Images
Figure CN116622508B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of cell bioreactor technology, and in particular to a disposable stirred tank cell bioreactor. Background Technology
[0002] Currently, significant amounts of capital, human resources, and technology are being invested globally in the research, development, capacity expansion, and industrialization of various vaccines, diagnostic reagents, antibody drugs, recombinant protein drugs, and cell therapy drugs. Since these biopharmaceuticals are all expressed using mammalian cells, the single-use stirred-tank cell bioreactor—the most crucial upstream bioprocess equipment in the biopharmaceutical industry for large-scale animal cell culture—has become a key factor and bottleneck in expanding production capacity. This cell bioreactor largely determines a biopharmaceutical company's production capacity, product variety, and product cost. Currently, single-use stirred-tank cell bioreactors are widely used in biopharmaceutical companies and are the most widely applied and mature type of cell bioreactor.
[0003] However, many disposable stirred tank cell bioreactors used by biopharmaceutical companies have numerous defects and shortcomings. For example: 1. Some reactors have top-entry or bottom-entry stirring shafts. Although the bearings inside the cell culture bag have sealing components, they cannot achieve a complete seal, and leaks still occur. Such reactors are not reliable enough to prevent contamination. 2. Some reactors meet the above conditions by using magnetic drive technology outside the culture bag to achieve a fully enclosed culture bag. However, the stirring shaft entering the bag is only a short shaft, which can only install one impeller, either at the bottom or the upper half of the culture bag. As a result, the culture medium cannot be uniformly stirred and an ideal flow field cannot be formed throughout the entire height of the culture bag, and the culture medium far from the impeller is not well stirred and mixed. At the same time, it reduces the dynamic gas-liquid specific surface area, affecting mass transfer. This also reduces the oxygen level of the culture medium and the escape rate of harmful gases, causing excessive retention of harmful gases such as carbon dioxide and ammonia in the culture medium. This results in reduced cell density and poor cell activity.
[0004] On the other hand, when the bubbles from the bubble ring at the bottom of the culture bag rise to the upper part of the bag, the poor flow of the culture medium causes the bubbles to escape only from the center of the culture medium. This lack of effective dissolved oxygen in the rest of the bag leads to insufficient dissolved oxygen in the culture solution, cell death, reduced cell density, and poor cell viability.
[0005] The information disclosed in this background section is intended only to enhance the understanding of the general background of the invention and should not be construed as an admission or in any way implying that the information constitutes prior art known to those skilled in the art. Summary of the Invention
[0006] The purpose of this invention is to provide a disposable stirred tank cell bioreactor to solve the technical problems existing in the prior art.
[0007] To achieve the above objectives, the present invention adopts the following technical solution:
[0008] This invention provides a disposable stirred tank cell bioreactor, comprising: a positioning short shaft clamping assembly, a cell culture bag assembly, a stirring shaft rotating assembly, a reaction tank body, and a drive mechanism assembly; the positioning short shaft clamping assembly is installed on one side of the upper part of the reaction tank body; the stirring shaft rotating assembly is installed inside the cell culture bag assembly, and is located in the space above the culture medium surface and below the top of the cell culture bag assembly when the cell culture bag assembly is full of working volume; the stirring shaft rotating assembly is located inside the cell culture bag assembly, and its upper end is connected to the positioning short shaft clamping assembly; the lower end of the stirring shaft rotating assembly is connected to the drive mechanism assembly; the cell culture bag assembly is installed inside the reaction tank body.
[0009] Preferably, the positioning short shaft clamping assembly includes: a positioning short shaft, a V-shaped chuck, a movable V-shaped chuck, a fastening bolt, a positioning channel steel, a hinge shaft, an arc-shaped support plate, a flip-plate shaft, a flip-up triangular plate, a clamping nut, a flip-plate fastening bolt, and a clamping bolt; the positioning short shaft is clamped and positioned by the V-shaped chuck and the movable V-shaped chuck; one side of the movable V-shaped chuck is connected to the V-shaped chuck via the hinge shaft, and the other side of the movable V-shaped chuck is clamped to the V-shaped chuck via the clamping bolt; the movable V-shaped chuck can bend around the clamp after the clamping bolt is released. The page shaft rotates so that the movable V-shaped chuck can form a preset opening angle (preferably, the opening angle of the movable V-shaped chuck is 146°); the V-shaped chuck is mounted on the rotatable triangular plate by the fastening bolt, the rotatable triangular plate is connected to the arc-shaped support plate by the flip-plate shaft, the rotatable triangular plate can rotate around the flip-plate shaft, and after completing the rotation action, the rotatable triangular plate can be fastened to the arc-shaped support plate by the flip-plate fastening bolt, and the rotation angle of the rotatable triangular plate is 100°.
[0010] Preferably, one end of the clamping bolt is pivotally connected to the side wall of the V-shaped chuck, and the movable V-shaped chuck is correspondingly provided with a side opening for mounting the other end of the clamping bolt. (The opening angle of the clamping bolt is 55°).
[0011] Preferably, a long fastening bolt is installed through the interior of the positioning short shaft.
[0012] Preferably, the cell culture bag assembly includes: a large-diameter air inlet, a pressure-resistant air inlet hose, a bubble ring, multiple rigid plastic interfaces, a cell culture bag, a rigid plastic sealing cover, and sensor elements; the large-diameter air inlet is welded to the top of the cell culture bag, and its lower end is connected to the pressure-resistant air inlet hose; the pressure-resistant air inlet hose extends downward to the bottom of the cell culture bag and connects to the port of the bubble ring; the rigid plastic sealing cover is sealed and welded to the bottom of the cell culture bag; the multiple rigid plastic interfaces are welded to the top of the cell culture bag; and the sensor elements (including pH, dissolved oxygen, and other sensors) are sealed and welded to the lower part of the cell culture bag.
[0013] Preferably, the rotating assembly of the stirring shaft includes: a bushing of a straight shaft, a shaped silicone hose, a plastic strap, a rigid plastic tube for connecting the culture bag, a condensate-blocking sealing ring, two thrust bearing bushings, two thrust bearings, a sliding bearing bushing, a plastic sliding bearing, an end cap, a connecting pin, a rigid plastic stirring shaft, a rigid plastic sealing cup, a rigid plastic sealing cup lid, an internal hex bolt, a spring washer, a thrust bushing, a rotating shaft, and multiple bolts; the bushing of the straight shaft is fixedly connected to the positioning short shaft; the long fastening bolt passes through the positioning short shaft and is threaded to the shaft of the straight shaft. The bottom of the sleeve; the bushing of the straight shaft is sealed to the upper end of the shaped silicone tubing via the plastic strap; the lower end of the shaped silicone tubing is installed at the upper end of the rigid plastic tube opening connecting the culture bag via the plastic strap, the lower end of the rigid plastic tube opening connecting the culture bag is sealed and welded to the top of the cell culture bag, the lower end of the bushing of the straight shaft is connected to the upper end of the two thrust bearing bushings via the plurality of bolts, the two thrust bearings are installed between the two thrust bearing bushings and the rotating shaft, and the thrust bushing is installed on the rotating shaft. The top of the shaft is secured to the rotating shaft by the hexagonal socket head cap screw and the spring washer; the thrust bushing is pressed against the inner ring of the two thrust bearings; the lower ends of the two thrust bearing bushings are coaxially connected to the upper end of the sliding bearing bushing by multiple bolts; the plastic sliding bearing is installed between the sliding bearing bushing and the rotating shaft; the lower end of the sliding bearing bushing is connected to an end cap by the multiple bolts to prevent the plastic sliding bearing from sliding axially; the lower end of the rotating shaft is coaxially connected to the hard plastic stirring shaft by two connecting pins. The lower end of the plastic sealing cup is sealed and welded to the outer surface of the hard plastic stirring shaft. The upper end of the hard plastic sealing cup is coaxially threaded to the hard plastic sealing cup cover. The condensate-blocking sealing ring is tightly fixed to the lower part of the bushing of the straight shaft by the plastic strap and is located inside the cell culture bag and between the hard plastic sealing cup cover. The hard plastic stirring shaft is equipped with an upper hard plastic stirring paddle and a lower hard plastic stirring paddle. A radial magnetic coupler driven bushing is installed at the lower end of the hard plastic stirring shaft, and the radial magnetic coupler driven bushing is coaxially fitted around the hard plastic sealing cover.
[0014] Preferably, the top end of the bushing of the through shaft is provided with a positioning groove on the bushing; a positioning channel steel is provided below the V-shaped chuck; the positioning groove on the bushing is an annular groove on the outer shaft surface of the top end of the bushing of the through shaft, and the annular groove is inserted into the positioning channel steel to position the cell culture bag assembly when installing the cell culture bag.
[0015] Preferably, the reaction vessel includes: a stainless steel vessel, a vessel door, a sensor window, a vessel support, an adjustable support plate, a heating water jacket, a vessel door locking mechanism, and a door hinge shaft; the cell culture bag is installed inside the stainless steel vessel; the stainless steel vessel is welded to the vessel support; the adjustable support plate is installed on the bottom plate of the vessel support; the vessel door is connected to the stainless steel vessel by the door hinge shaft; the vessel door has an opening angle; when the vessel door is closed, it is locked by the vessel door locking mechanism; the stainless steel vessel is equipped with the heating water jacket to heat and keep the culture medium warm; the positioning short shaft clamping assembly is connected to the stainless steel vessel through the arc-shaped support plate thereon; and the sensor window is located on the lower door side of the stainless steel vessel.
[0016] Preferably, the drive mechanism assembly includes: a radial magnetic coupler drive shaft and a motor; the radial magnetic coupler drive shaft and the motor are installed at the center of the bottom of the stainless steel tank, and the motor drives the radial magnetic coupler drive shaft to rotate; the radial magnetic coupler drive shaft cooperates with the radial magnetic coupler driven bushing to achieve magnetic transmission through magnetic coupling, thereby causing the radial magnetic coupler driven bushing to rotate.
[0017] Preferably, both the radial magnetic coupler driven bushing and the cell culture bag are single-use products. The radial magnetic coupler drive shaft and the motor are reusable products.
[0018] By adopting the above technical solution, the present invention has the following beneficial effects:
[0019] The disposable stirred tank cell bioreactor provided by this invention is highly innovative, has high reliability, and can be applied to cell culture processes.
[0020] Furthermore, the present invention, through the combination of the positioning short shaft clamping assembly, the cell culture bag assembly, the stirring shaft rotating assembly, the reaction vessel body, and the drive mechanism assembly, has the following advantages:
[0021] (a) Advantages of the positioning short shaft clamping assembly:
[0022] (1) When the V-shaped chuck and the movable V-shaped chuck of the present invention clamp the positioning short shaft, the two V-shaped chucks clamp each other, which can automatically fix the center of the positioning short shaft. After clamping and fixing, the positioning short shaft can be fixed in the vertical direction of the axial direction and bear the upward and downward forces along the axial direction.
[0023] (2) Both the V-shaped clamp and the movable V-shaped clamp of this invention are fixed on the rotatable triangular plate, and the rotatable triangular plate can rotate up to 100 degrees. When rotated to the maximum angle, the top of the stainless steel reaction vessel is completely empty. At this time, it is very convenient to install cell culture bags into the vessel. Large-volume cell culture bags can also be accurately and quickly installed into the reaction vessel from top to bottom, and the empty culture bag can be easily removed from the top of the reaction vessel at the end of cell culture, and a new cell culture bag can be put in for the next cell culture process.
[0024] (II) Advantages of the stirring shaft rotating assembly:
[0025] (1) The stirring shaft rotating assembly of the present invention is placed in the upper part of the cell culture bag, above the full volume liquid surface and below the top of the cell culture bag, in a gaseous space, without contacting the culture medium and thus not contaminating it. The outer circle of the bushing of the straight shaft at the upper end of the stirring shaft rotating assembly is tightly bound to the shaped silicone tubing with plastic straps, ensuring a completely sealed state inside the bag. The lower end of the shaped silicone tubing is tightly sealed to the upper end of the hard plastic tube connecting the culture bag with plastic straps, preventing gas from outside the cell culture bag from entering and forming a perfect seal. The positioning groove on the bushing at the top of the stirring shaft rotating assembly is an annular groove on the outer shaft surface of the bushing top of the straight shaft. When installing the cell culture bag into the stainless steel reaction vessel, the annular groove is inserted into the positioning channel steel to position the cell culture bag assembly. The positioning groove on the bushing is an annular groove on the outer shaft surface of the bushing top of the straight shaft. When installing the cell culture bag, this annular groove will be inserted into the positioning channel steel for positioning.
[0026] (2) In the inner part of the stirring shaft rotating assembly of the present invention, the rotation transmitted from the hard plastic stirring shaft to the rotating shaft stops after reaching the top of the rotating shaft and the hard plastic sealing cup lid. The bushing of the straight shaft at the top of the assembly is stationary. The rotational motion transmitted from the hard plastic stirring shaft to the rotating shaft is supported and completed by the plastic sliding bearing and the sliding bearing bushing. The axial force transmitted simultaneously with the rotation of the rotating shaft is supported and completed by the two thrust bearings and the two thrust bearing bushings.
[0027] (3) A hard plastic sealing cup is welded onto the hard plastic stirring shaft of this invention, and a hard plastic sealing cup lid is threaded onto it. Both are connected to the hard plastic stirring shaft and rotate together. The condensate-blocking sealing ring is fixed to the bushing of the straight shaft and remains stationary. The function of the hard plastic sealing cup, the cup lid, and the condensate-blocking sealing ring is to prevent condensate and water vapor from entering the sealing cup and contacting the bearings and bearing sleeves inside.
[0028] (4) In summary, the beneficial effects of the stirring shaft rotating assembly are that the entire rotating part of the upper part of the stirring shaft is within the cell culture area. The top of the cell culture bag has a stationary straight shaft sleeve that is tightly sealed with a shaped silicone tube, which achieves a complete seal of the straight shaft sleeve at the top outlet of the cell culture bag.
[0029] (III) Advantages of the cell culture bag assembly:
[0030] (1) The bottom of the hard plastic sealing cover at the bottom of the cell culture bag is directly sealed and welded to the plastic film at the bottom of the culture bag, which achieves a complete seal at the bottom of the cell.
[0031] (2) The driven shaft sleeve of the radial magnetic coupler fixed at the lower end of the rigid plastic stirring shaft inside the bag is engaged with the drive shaft head of the radial magnetic coupler installed at the bottom of the stainless steel tank outside the bag. Through magnetic coupling, the driven shaft sleeve of the radial magnetic coupler inside the bag is rotated by magnetic drive when the bag is sealed, thereby causing the rigid plastic stirring shaft to rotate and driving the two rigid plastic stirring paddles to complete the stirring of the culture medium.
[0032] (3) Due to the sealing and welding of the various hard plastic interfaces, large-diameter air inlets and interfaces of sensors such as pH and dissolved oxygen on the cell culture bag and the plastic film of the culture bag, the interfaces of these interfaces are perfectly sealed.
[0033] (4) The straight-through shaft sleeve extending from the top of the stirring shaft to the inside of the cell culture bag and the magnetic drive component at the bottom of the cell culture bag forms the entire top-to-bottom straight-through stirring shaft. This straight-through stirring shaft can be equipped with multiple stirring paddles of various structures to achieve stirring of the culture medium at different heights in the culture bag, so that the culture medium forms the required flow state and flow field. This is more conducive to cell growth and expansion. On the other hand, this straight-through stirring shaft can improve the overall height-to-diameter ratio of the disposable stirred tank cell bioreactor - that is, the cell reactor is thinner and taller. This can greatly improve the adjustment ratio of the cell reactor. Thus, the number of stages in the stepwise cell culture process is reduced. This also reduces the number of equipment such as cell bioreactors, and the number of times the cell culture tank is turned over for expansion in the stepwise cell culture process, thus reducing the risk of contamination in the cell culture tank turning over process. At the same time, it also reduces labor and material costs and shortens the production cycle of the entire cell culture process. Attached Figure Description
[0034] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.
[0035] Figure 1 A cross-sectional view of the positioning short shaft clamping assembly provided in an embodiment of the present invention;
[0036] Figure 2 A top view of the positioning short shaft clamping assembly provided in an embodiment of the present invention;
[0037] Figure 3 A side view of the positioning short shaft clamping assembly provided in an embodiment of the present invention;
[0038] Figure 4 This is a front view of the cell culture bag assembly provided in an embodiment of the present invention;
[0039] Figure 5 This is a side view of the cell culture bag assembly provided in an embodiment of the present invention;
[0040] Figure 6 This is a top view of the cell culture bag assembly provided in an embodiment of the present invention;
[0041] Figure 7 This is a top sectional view of the stirring shaft rotating assembly provided in an embodiment of the present invention;
[0042] Figure 8 This is a front view of the reaction vessel provided in an embodiment of the present invention;
[0043] Figure 9 A side view of the reaction vessel provided in an embodiment of the present invention;
[0044] Figure 10 A top view of the reaction vessel provided in an embodiment of the present invention;
[0045] Figure 11 This is a schematic diagram of the structure of a disposable stirred tank cell bioreactor provided in an embodiment of the present invention;
[0046] Figure 12 A top view of a disposable stirred tank cell bioreactor provided in an embodiment of the present invention;
[0047] Figure 13 A side view of a disposable stirred tank cell bioreactor provided in an embodiment of the present invention. Detailed Implementation
[0048] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0049] The specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are for illustration and explanation only and are not intended to limit the present invention.
[0050] Combination Figures 1 to 13 As shown, this embodiment provides a disposable stirred tank cell bioreactor, comprising: a positioning short shaft clamping assembly 300, a cell culture bag assembly 200, a stirring shaft rotating assembly 100, a reaction tank 400, and a drive mechanism assembly; the positioning short shaft clamping assembly 300 is installed on one side of the upper part of the reaction tank 400; the stirring shaft rotating assembly 100 is installed inside the cell culture bag assembly 200, and is located in the space above the culture medium surface and below the top of the cell culture bag assembly 200 when the cell culture bag assembly 200 is full; the stirring shaft rotating assembly 100 is located inside the cell culture bag assembly 200, and its upper end is connected to the positioning short shaft clamping assembly; the lower end of the stirring shaft rotating assembly 100 is connected to the drive mechanism assembly; the cell culture bag assembly 200 is installed inside the reaction tank 400. The disposable stirred tank cell bioreactor provided in this embodiment is highly innovative, has high reliability, and can be applied to cell culture processes.
[0051] In this embodiment, preferably, the positioning short shaft clamping assembly 300 includes: a positioning short shaft 311, a V-shaped chuck 321, a movable V-shaped chuck 322, a fastening bolt 372, a positioning channel steel 332, a hinge shaft 326, an arc-shaped support plate 362, a flip-plate shaft 343, a flip-up triangular plate 342, a clamping nut, a flip-plate fastening bolt 348, and a clamping bolt 325; the positioning short shaft 311 is clamped and positioned by the V-shaped chuck 321 and the movable V-shaped chuck 322; one side of the movable V-shaped chuck 322 is connected to the V-shaped chuck 321 via the hinge shaft 326, and the other side of the movable V-shaped chuck 322 is clamped to the V-shaped chuck 321 via the clamping bolt 325; the movable V-shaped chuck 322... After the clamping bolt 325 is loosened, the chuck 322 can rotate around the folding shaft 326 so that the movable V-shaped chuck 322 can form a preset opening angle. Preferably, the opening angle of the movable V-shaped chuck is 146°. The V-shaped chuck 321 is mounted on the flip-out triangular plate 342 by the fastening bolt 372. The flip-out triangular plate 342 is connected to the arc-shaped support plate 362 by the flipping shaft 343. The flip-out triangular plate 342 can rotate around the flipping shaft 343. After the flip-out action is completed, the flip-out triangular plate 342 can be fastened to the arc-shaped support plate 362 by the flipping fastening bolt 348. The rotation angle of the flip-out triangular plate 342 is 100°.
[0052] In this embodiment, preferably, one end of the clamping bolt 325 is pivotally connected to the side wall of the V-shaped chuck 321, and the movable V-shaped chuck 322 is correspondingly provided with a side opening for mounting the other end of the clamping bolt 325. The opening angle of the clamping bolt 325 is 55°.
[0053] In this embodiment, preferably, a long fastening bolt 314 is installed through the interior of the positioning short shaft 311.
[0054] In this embodiment, preferably, the cell culture bag assembly 200 includes: a large-diameter air inlet 212, a pressure-resistant air inlet hose 214, a bubble ring 218, various rigid plastic interfaces 222, a cell culture bag 201, a rigid plastic sealing cover 239, and a sensor element 252; the large-diameter air inlet 212 is welded to the top of the cell culture bag 201, and its lower end is connected to the pressure-resistant air inlet hose 214; the pressure-resistant air inlet hose 214 extends downward to the bottom of the cell culture bag 201 and connects to the port of the bubble ring 218; the rigid plastic sealing cover 239 is sealed and welded to the bottom of the cell culture bag 201; the various rigid plastic interfaces 222 are welded to the top of the cell culture bag 201; and the sensor element 252 is sealed and welded to the lower part of the cell culture bag 201. The sensor element 252 includes sensors for pH, dissolved oxygen, etc.
[0055] In this embodiment, preferably, the stirring shaft rotating assembly 100 includes: a bushing 111 for a straight shaft, a shaped silicone hose 126, a plastic strap 121, a rigid plastic tube opening 128 for connecting the culture bag, a condensate-blocking sealing ring 142, two thrust bearing bushings 114, two thrust bearings 162, a sliding bearing bushing 116, a plastic sliding bearing 164, an end cap 117, a connecting pin 169, a rigid plastic stirring shaft 232, a rigid plastic sealing cup 154, a rigid plastic sealing cup cover 151, an internal hex bolt 161, a spring washer 163, a thrust bushing 166, a rotating shaft 168, and multiple bolts 141; the bushing 111 of the straight shaft and the positioning short shaft 31 1. Fixed connection; a long fastening bolt 314 passes through the positioning short shaft 311 and is threaded to the bottom of the bushing 111 of the straight shaft; the bushing 111 of the straight shaft is sealed to the upper end of the shaped silicone tube 126 via a plastic strap 121; the lower end of the shaped silicone tube 126 is installed at the upper end of the rigid plastic tube opening 128 connecting the culture bag via a plastic strap 121; the lower end of the rigid plastic tube opening 128 connecting the culture bag is sealed and welded to the top of the cell culture bag 201; the lower end of the bushing 111 of the straight shaft is connected to the upper end of the two thrust bearing bushings 114 via multiple bolts; the two thrust bearings 162 are installed between the two thrust bearing bushings 114 and the rotating shaft 168. A thrust sleeve 166 is installed on the top of the rotating shaft 168 and fastened to the top of the rotating shaft 168 by an internal hex bolt 161 and a spring washer 163. The thrust sleeve 166 is pressed against the inner ring of the two thrust bearings 162. The lower ends of the two thrust bearing sleeves 114 are coaxially connected to the upper ends of the sliding bearing sleeves 116 by multiple bolts 141. A plastic sliding bearing 164 is installed between the sliding bearing sleeves 116 and the rotating shaft 168. The lower end of the sliding bearing sleeves 116 is connected to an end cap 117 by multiple bolts to prevent the plastic sliding bearing 164 from sliding axially. The lower end of the rotating shaft 168 is coaxially connected to the hard plastic stirring shaft 232 by two connecting pins 169. The rigid plastic sealing cup 154 is welded to the outer surface of the rigid plastic stirring shaft 232 at its lower end. The upper end of the rigid plastic sealing cup 154 is threadedly connected to the rigid plastic sealing cup cover 151 on the same axis. The condensate-blocking sealing ring 142 is secured to the lower part of the bushing 111 of the straight shaft by plastic straps 121 and is located inside the cell culture bag 201 and between the rigid plastic sealing cup cover 151. The rigid plastic stirring shaft 232 is equipped with an upper rigid plastic stirring paddle 241 and a lower rigid plastic stirring paddle 242. The lower end of the rigid plastic stirring shaft 232 is equipped with a radial magnetic coupler driven bushing 281, which is coaxially fitted around the rigid plastic sealing cover 239.
[0056] In this embodiment, preferably, the top end of the bushing 111 of the straight shaft is provided with a bushing positioning groove 112; a positioning channel steel 332 is provided below the V-shaped chuck 321; the bushing positioning groove 112 is an annular groove on the outer shaft surface of the top end of the bushing 111 of the straight shaft, and when installing the cell culture bag 201, the annular groove is inserted into the positioning channel steel 332 to position the cell culture bag assembly 200.
[0057] In this embodiment, preferably, the reaction vessel 400 includes: a stainless steel vessel 412, a vessel door 422, a sensor window 432, a vessel support 442, an adjustable support plate 452, a heating water jacket 416, a vessel door locking mechanism 426, and a door hinge shaft 462; the cell culture bag 201 is installed inside the stainless steel vessel 412; the stainless steel vessel 412 is welded to the vessel support 442; the adjustable support plate 452 is installed on the bottom plate of the vessel support 442, and the vessel... The body door 422 is connected to the stainless steel tank 412 by a door hinge shaft 462. The tank door 422 is provided with an opening angle. When the tank door 422 is closed, it is locked by the tank door locking mechanism 426. The stainless steel tank 412 is equipped with a heating water jacket 416 to heat and keep the culture medium warm. The positioning short shaft clamping assembly 300 is connected to the stainless steel tank 412 through the arc-shaped support plate 362 on it. The sensor window 432 is opened on the lower door side of the stainless steel tank 412.
[0058] In this embodiment, preferably, the drive mechanism assembly includes: a radial magnetic coupler drive shaft head 502 and a motor 508; the radial magnetic coupler drive shaft head 502 and the motor 508 are installed at the center of the bottom of the stainless steel tank 412, and the motor 508 drives the radial magnetic coupler drive shaft head 502 to rotate; the radial magnetic coupler drive shaft head 502 is used in conjunction with the radial magnetic coupler driven shaft sleeve 281 to achieve magnetic transmission through magnetic coupling, thereby causing the radial magnetic coupler driven shaft sleeve 281 to rotate.
[0059] In this embodiment, preferably, both the radial magnetic coupler driven bushing 281 and the cell culture bag 201 are single-use products. The radial magnetic coupler drive shaft head 502 and the motor 508 are reusable products.
[0060] It is worth noting that the disposable stirred tank cell bioreactor of this application has the following innovative features:
[0061] The first innovation is the seal between the upper straight shaft sleeve and the shaped silicone tubing, achieved through a strap. This seal connects the straight shaft exiting the culture bag to the culture bag itself. There's also a tight seal at the top of the hard plastic tube opening and the bottom of the shaped silicone tubing. The second innovation is the stirring shaft rotating assembly. It's located in the air-filled space between the liquid level when the culture bag is full and the top of the bag, not in the liquid itself. Therefore, its placement is innovative. The rotational torque transmitted from the bottom of the stirring shaft is transferred to the rotating assembly via a plastic sliding bearing, supported by a sliding bearing sleeve. The upward and downward axial forces are borne by two thrust bearings and their sleeves. The straight shaft sleeve at the top of the rotating assembly remains stationary.
[0062] The second innovation is twofold: First, the clamping and positioning mechanism of the V-shaped gripper. Second, the flip-up triangular plate has an upward rotation angle of 100 degrees. When the reactor is operating, this plate is placed horizontally; when installing cell culture bags, it is flipped upwards. This facilitates the installation and removal of cell culture bags.
[0063] The third innovation is the inclusion of a straight-through shaft within the cell culture bag. This shaft extends radially from the bottom of the bag's interior, via a magnetic coupler, to the top of the straight-through shaft. Combined with the fully sealed cell culture bag, this constitutes the straight-through shaft. The advantages of this shaft are twofold: First, it allows for the installation of two or more agitators vertically to the shaft. The placement of each agitator can be determined based on the required flow field of the culture medium at various points within the bag. This enables control over the flow field throughout the cell culture bag. Second, it allows for the construction of a taller, narrower reactor with a larger height-to-diameter ratio. Such a reactor offers a greater adjustment ratio. The adjustment ratio is the ratio of the reactor's maximum working volume to its minimum working volume. The larger the difference between the maximum and minimum working volumes, the greater the adjustment ratio, and vice versa. For example, a reactor with a maximum working volume of 1000 liters has a minimum working volume of 500 liters, resulting in an adjustment ratio of 2:1. In this case, a reactor with a volume smaller than 500 liters cannot be used for cell culture; a smaller reactor must be found. This would require using an additional reactor stage. If the minimum working volume of the reactor is 100 liters, the adjustment ratio is 10:1. In this case, the reactor can be used for cultivation within a volume range of 100-1000 liters. This reduces the need for a single reactor stage. This is the significance of the adjustment ratio. Because of its narrow profile, this type of reactor also has the advantage of agitating the culture medium inside the culture bag near the tank wall, preventing flow lag or stagnation.
[0064] It is understandable that: firstly, the radial magnetic coupler at the bottom of the reaction vessel and its matching motor are all purchased externally; secondly, some components for installing sensors such as pH and dissolved oxygen are also purchased externally, allowing for flexible selection based on actual needs.
[0065] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.
Claims
1. A disposable stirred tank cell bioreactor, characterized in that, include: The reactor comprises a positioning short shaft clamping assembly (300), a cell culture bag assembly (200), a stirring shaft rotating assembly (100), a reaction vessel (400), and a drive mechanism assembly; the positioning short shaft clamping assembly (300) is installed on one side of the upper part of the reaction vessel (400); the stirring shaft rotating assembly (100) is installed inside the cell culture bag assembly (200) and is located in the space above the culture medium surface and below the top of the cell culture bag assembly (200) when the cell culture bag assembly (200) is full; the upper end of the stirring shaft rotating assembly (100) is connected to the positioning short shaft clamping assembly; the lower end of the stirring shaft rotating assembly (100) is connected to the drive mechanism assembly; and the cell culture bag assembly (200) is installed inside the reaction vessel (400). The positioning short shaft clamping assembly (300) includes: a positioning short shaft (311), a V-shaped chuck (321), a movable V-shaped chuck (322), a fastening bolt (372), a positioning channel steel (332), a hinge shaft (326), an arc-shaped support plate (362), a flip-plate shaft (343), a flip-up triangular plate (342), a clamping nut, a flip-up fastening bolt (348), and a clamping bolt (325); the positioning short shaft (311) is clamped and positioned by the V-shaped chuck (321) and the movable V-shaped chuck (322); one side of the movable V-shaped chuck (322) is connected to the V-shaped chuck (321) through the hinge shaft (326), and the movable V-shaped chuck (311) is clamped and positioned by the V-shaped chuck (321) and the movable V-shaped chuck (322). The other side of the 322) is clamped to the V-shaped chuck (321) by the clamping bolt (325); the V-shaped chuck (321) is mounted on the reversible triangular plate (342) by the fastening bolt (372); the reversible triangular plate (342) is connected to the arc-shaped support plate (362) by the flipping shaft (343); the reversible triangular plate (342) can rotate around the flipping shaft (343); after the reversible triangular plate (342) has completed the rotation action, it can be fastened to the arc-shaped support plate (362) by the flipping fastening bolt (348); the rotation angle of the reversible triangular plate (342) is 100°. The rotating assembly (100) of the stirring shaft includes: a bushing (111) of a straight shaft, a shaped silicone tube (126), a plastic strap (121), a rigid plastic tube opening (128) for connecting the culture bag, two thrust bearing bushings (114), a sliding bearing bushing (116), a plastic sliding bearing (164), two thrust bearings (162), a thrust bushing (166), a rotating shaft (168), an internal hex bolt (161), a spring washer (163), and multiple bolts (141); the lower end of the bushing (111) of the straight shaft is connected to the upper end of the two thrust bearing bushings (114) through the multiple bolts, and the two thrust bearings (162) The thrust bushing (166) is installed between the two thrust bearing bushings (114) and the rotating shaft (168). The thrust bushing (166) is installed on the top of the rotating shaft (168) and is fastened to the top of the rotating shaft (168) by an internal hex bolt (161) and a spring washer (163). The thrust bushing (166) is pressed against the inner ring of the two thrust bearings (162). The lower end of the two thrust bearing bushings (114) is coaxially connected to the upper end of the sliding bearing bushing (116) by multiple bolts (141). The plastic sliding bearing (164) is installed between the sliding bearing bushing (116) and the rotating shaft (168).
2. The disposable stirred tank cell bioreactor according to claim 1, characterized in that, One end of the clamping bolt (325) is pivotally connected to the side wall of the V-shaped chuck (321), and the movable V-shaped chuck (322) is provided with a side opening for installing the other end of the clamping bolt (325).
3. The disposable stirred tank cell bioreactor according to claim 2, characterized in that, The positioning short shaft (311) is internally fitted with a long fastening bolt (314).
4. The disposable stirred tank cell bioreactor according to claim 3, characterized in that, The cell culture bag assembly (200) includes: a large-diameter air inlet (212), a pressure-resistant air inlet hose (214), a bubble ring (218), multiple hard plastic interfaces (222), a cell culture bag (201), a hard plastic sealing cover (239), and a sensor element (252); the large-diameter air inlet (212) is welded to the top of the cell culture bag (201), and its lower port is connected to the pressure-resistant air inlet hose (214); the pressure-resistant air inlet hose (214) extends downward to the bottom of the cell culture bag (201) and is connected to the port of the bubble ring (218); the hard plastic sealing cover (239) is sealed and welded to the bottom of the cell culture bag (201); the multiple hard plastic interfaces (222) are welded to the top of the cell culture bag (201); and the sensor element (252) is sealed and welded to the lower part of the cell culture bag (201).
5. The disposable stirred tank cell bioreactor according to claim 4, characterized in that, The stirring shaft rotating assembly (100) includes: a condensate-blocking sealing ring (142), an end cap (117), a connecting pin (169), a hard plastic stirring shaft (232), a hard plastic sealing cup (154), and a hard plastic sealing cup lid (151). The bushing (111) of the straight shaft is fixedly connected to the positioning short shaft (311); the long fastening bolt (314) passes through the positioning short shaft (311) and is threaded to the bottom of the bushing (111) of the straight shaft; the bushing (111) of the straight shaft is sealed to the upper end of the shaped silicone tube (126) through the plastic strap (121); the lower end of the shaped silicone tube (126) is installed at the upper end of the rigid plastic tube opening (128) of the connecting culture bag through the plastic strap (121), the lower end of the rigid plastic tube opening (128) of the connecting culture bag is sealed and welded to the top of the cell culture bag (201), the lower end of the sliding bearing bushing (116) is connected to the end cap (117) through the multiple bolts, and the lower end of the rotating shaft (168) is connected to the rigid plastic tube opening (169) through two connecting pins (169). The stirring shaft (232) is coaxially connected, and the lower end of the hard plastic sealing cup (154) is sealed and welded to the outer surface of the hard plastic stirring shaft (232). The upper end of the hard plastic sealing cup (154) is coaxially threaded to the hard plastic sealing cup cover (151). The condensate-blocking sealing ring (142) is tightly fixed to the lower part of the bushing (111) of the straight shaft by the plastic strap (121) and is located inside the cell culture bag (201) and between the hard plastic sealing cup cover (151). The hard plastic stirring shaft (232) is equipped with an upper hard plastic stirring paddle (241) and a lower hard plastic stirring paddle (242). The lower end of the hard plastic stirring shaft (232) is equipped with a radial magnetic coupler driven bushing (281), which is coaxially fitted around the hard plastic sealing cover (239).
6. The disposable stirred tank cell bioreactor according to claim 5, characterized in that, The bushing (111) of the straight shaft is provided with a bushing positioning groove (112) at the top end; a positioning channel steel (332) is provided below the V-shaped chuck (321); the bushing positioning groove (112) is an annular groove on the outer shaft surface of the top end of the bushing (111) of the straight shaft, and the annular groove is inserted into the positioning channel steel (332) to position the cell culture bag assembly (200) when installing the cell culture bag (201).
7. The disposable stirred tank cell bioreactor according to claim 6, characterized in that, The reaction vessel (400) includes: a stainless steel vessel (412), a vessel door (422), a sensor window (432), a vessel support (442), an adjustable support plate (452), a heating water jacket (416), a vessel door locking mechanism (426), and a door hinge shaft (462). The cell culture bag (201) is installed inside the stainless steel tank (412); the stainless steel tank (412) is welded to the tank support (442); the adjustable support plate (452) is installed on the bottom plate of the tank support (442); the tank door (422) is connected to the stainless steel tank (412) by the door hinge shaft (462); the tank door (422) is provided with an opening angle; when the tank door (422) is closed, it is locked by the tank door locking mechanism (426); the stainless steel tank (412) is equipped with the heating water jacket (416) to heat and keep the culture medium warm; the positioning short shaft clamping assembly (300) is connected to the stainless steel tank (412) through the arc-shaped support plate (362) thereon; the sensor window (432) is opened on the lower door side of the stainless steel tank (412).
8. The disposable stirred tank cell bioreactor according to claim 7, characterized in that, The drive mechanism assembly includes: a radial magnetic coupler drive shaft head (502) and a motor (508). The radial magnetic coupler drive shaft head (502) and the motor (508) are installed at the center of the bottom of the stainless steel tank (412). The motor (508) drives the radial magnetic coupler drive shaft head (502) to rotate. The radial magnetic coupler drive shaft head (502) is used in conjunction with the radial magnetic coupler driven shaft sleeve (281) to achieve magnetic transmission through magnetic coupling, thereby causing the radial magnetic coupler driven shaft sleeve (281) to rotate.
9. The disposable stirred tank cell bioreactor according to claim 8, characterized in that, Both the radial magnetic coupler driven bushing (281) and the cell culture bag (201) are single-use products.