CN111321064A

CN111321064A - Bioreactor for preparing monoclonal antibody

Info

Publication number: CN111321064A
Application number: CN202010158313.5A
Authority: CN
Inventors: 雍金贵; 缪连军
Original assignee: Anhui Global Gene Technology Co ltd
Current assignee: Anhui Global Gene Technology Co ltd
Priority date: 2020-03-09
Filing date: 2020-03-09
Publication date: 2020-06-23

Abstract

The invention discloses a bioreactor for preparing monoclonal antibody, which comprises a tank body, wherein a reaction bin is arranged in the tank body, a first rotating shaft is arranged in the reaction bin, a plurality of stirring plates which are oppositely arranged are arranged on two sides of the first rotating shaft, a heater is fixed on one side of the outside of the tank body, a temperature sensor and a pH controller are fixed on the side wall of the reaction bin, the heater, the temperature sensor and the pH controller are controlled by a PLC (programmable logic controller), two defoaming plates which are oppositely arranged are fixed on the first rotating shaft, the defoaming plates are positioned above the stirring plates, a plurality of defoaming blocks are arranged on the lower surfaces of the defoaming plates, defoaming needles are fixed on the lower surfaces of the defoaming blocks, and an air inlet mechanism is arranged outside the reaction bin; through the defoaming needle can be fine when the hybridoma cell culture, the bubble of production is eliminated, and the stirring board has effectually prevented to produce the vortex flow at the stirring in-process, and the mechanism of admitting air is inside reaction bin with required gaseous homodisperse, and then has promoted monoclonal antibody's preparation efficiency.

Description

Bioreactor for preparing monoclonal antibody

Technical Field

The invention belongs to the field of monoclonal antibody preparation, and particularly relates to a bioreactor for preparing monoclonal antibody.

Background

The monoclonal antibody is a highly uniform antibody which is generated by a single B cell clone and only aims at a certain specific epitope, and is generally prepared by adopting a hybridoma technology, wherein the hybridoma technology is characterized in that sensitized B cells with the capacity of secreting specific antibodies and myeloma cells with the unlimited reproduction capacity are fused into B cell hybridomas on the basis of a cell fusion technology, so that the culture of the hybridoma cells is crucial in the process of preparing the monoclonal antibody;

in the process of preparing the large-scale monoclonal antibody, the culture of the hybridoma cells is usually carried out in vitro through a bioreactor, and when a stirring plate in the traditional bioreactor is used for stirring, although the rotating speed is slow, a swirling flow can be generated, the swirling flow can cause the axial circulating speed to be always lower than the radial circulating speed, the uniform dispersion of hybridoma cells is influenced, the violently swirling culture solution can cause waves which are repeatedly rushed, and impact force can be generated on equipment under the action of the swirling flow, so that the service life of the equipment is shortened, simultaneously to leading to the cell damage, and can produce the one deck bubble on the surface of culture solution in culture process, the production of bubble can cause a large amount of culture solution to escape the liquid, and the culture solution escapes from outlet duct or bearing seal and increases the chance of contracting bacteria, influences gaseous entering reaction storehouse when serious, makes the cell breathe and receives the hindrance, leads to the metabolism unusual, and then influences monoclonal antibody's preparation.

Disclosure of Invention

The invention aims to provide a bioreactor for preparing monoclonal antibody.

The technical problems to be solved by the invention are as follows:

1. in the process of preparing the monoclonal antibody on a large scale, a bioreactor is needed, a stirring plate in the traditional bioreactor generates a swirling flow although the rotating speed is slow when stirring is performed, the axial circulating speed is usually lower than the radial circulating speed due to the swirling flow, the uniform dispersion of hybridoma cells is influenced, the violently swirling culture solution can cause waves of reciprocating impulse, and in combination with the action of the swirling flow, impact force can be generated on equipment to further reduce the service life of the equipment, and meanwhile, the damage to cells is caused, so that the preparation of the monoclonal antibody is influenced;

2. in the process of cell culture, a layer of bubbles can be generated on the surface of a culture solution, the generation of the bubbles can cause a large amount of the culture solution to escape from the solution, the culture solution escapes from an air outlet pipe or a shaft seal to increase the chance of contamination, and gas is seriously influenced to enter a reaction bin, so that the cell breathing is hindered, and the metabolism is abnormal;

3. the traditional bioreactor needs to artificially detect and adjust the internal environment of the reaction bin, so that the culture efficiency of hybridoma cells is greatly reduced, and cell death is easily caused due to untimely culture environment, and the preparation of monoclonal antibodies is further influenced.

The purpose of the invention can be realized by the following technical scheme:

a bioreactor for preparing monoclonal antibody comprises a tank body, a reaction bin is arranged in the tank body, a first motor is fixed at the center of the top end of the outer portion of the tank body, a first rotating shaft is fixed on an output shaft of the first motor, the first rotating shaft sequentially penetrates through the side walls of the tank body and the reaction bin, the bottom end of the first rotating shaft is rotatably connected with the inner wall of the reaction bin, a plurality of stirring plates which are oppositely arranged are arranged on two sides of the first rotating shaft, a second motor is fixed at the center of the bottom end of the outer portion of the tank body, a second rotating shaft is fixed on an output shaft of the second motor, the second rotating shaft sequentially penetrates through the side walls of the tank body and the reaction bin, the second rotating shaft is rotatably connected with the side walls of the tank body and the reaction bin, the second rotating shaft is positioned in the first rotating shaft, the second rotating shaft is rotatably connected with the first rotating shaft, and a plurality of second main transmission gears which are uniformly distributed are fixed on the second rotating shaft, a first main transmission gear is also fixed on the second rotating shaft and is positioned above the second main transmission gear, a heater is fixed on one side outside the tank body, a temperature sensor and a pH controller are fixed on the side wall of the reaction bin, the heater, the temperature sensor and the pH controller are controlled by a PLC, a feed inlet is arranged at the top end of the tank body and is communicated with the inside of the reaction bin, a valve is arranged inside the feed inlet, a discharge end is arranged at the center of the bottom end outside the tank body, two defoaming plates which are oppositely arranged are fixed on the first rotating shaft and are positioned above the stirring plate, a plurality of defoaming blocks are arranged on the lower surfaces of the defoaming plates, defoaming needles are fixed on the lower surfaces of the defoaming blocks, an air inlet mechanism is arranged outside the reaction bin and is positioned between the reaction bin and the tank body, the air inlet mechanism comprises a vertical pipe, and the top end of air inlet penetrates through the side wall of, the lateral wall fixed connection of the standpipe and the jar body admits air, and the standpipe admits air one side that is close to the reaction chamber is fixed with a plurality of and admits air violently the pipe, admits air and violently manages to be fixed with first ring canal of giving vent to anger on the pipe, and first ring canal of giving vent to anger is located the inside of reaction chamber, and the top of reaction chamber is equipped with the outlet duct, and the outlet duct communicates with the reaction chamber is inside, and the.

Further, the outside bottom mounting of the jar body have four evenly distributed's support column, the bottom mounting of support column has the base, is fixed with the cooling bin on the outside lateral wall of the jar body, one side top in cooling bin is equipped with the inlet tube, the opposite side in cooling bin is equipped with the outlet pipe, inlet tube and outlet pipe respectively with the inside intercommunication in cooling bin, first pivot and second pivot antiport.

Further, the stirring board be located the inside in reaction bin, be fixed with the stirring curb plate that two opposition set up on the lateral wall of stirring board, the stirring curb plate is the arc board, the one end that the stirring board is close to first pivot is fixed with the stirring board pivot, the end of stirring board pivot is located the inside of first pivot, first pivot is connected with the rotation of stirring board pivot, be fixed with the stirring board gear on the end of stirring board pivot, the meshing of stirring board gear and second main drive gear mutually.

Further, a first fixing shaft is arranged at one end, close to the first rotating shaft, of the lower surface of the defoaming plate, the upper end of the first fixing shaft penetrates through the lower surface of the defoaming plate, the first fixing shaft is rotatably connected with the defoaming plate, a roller shaft gear is fixed at the upper end of the first fixing shaft, a second auxiliary transmission gear is vertically arranged on one side of the roller shaft gear and meshed with the roller shaft gear, a first connecting rod is fixed on the side wall of the second auxiliary transmission gear, a first auxiliary transmission gear is fixed at the end of the first connecting rod and meshed with the first main transmission gear, a first roller shaft is fixed at the lower end of the first fixing shaft, a second fixing shaft is fixed at one end, far away from the first rotating shaft, of the lower surface of the defoaming plate, a second roller shaft is arranged at the bottom end of the second fixing shaft and rotatably connected with the second fixing shaft, a transmission belt is arranged between the first roller shaft and the second roller, the lower surface of the defoaming block is fixed with a defoaming needle.

Further, the interior ring of the first ring canal of giving vent to anger in be equipped with the second ring canal of giving vent to anger, the lateral wall of the second ring canal of giving vent to anger is fixed with the connecting trachea of two opposition settings, connects tracheal end and the first ring canal fixed connection of giving vent to anger, the first ring canal of giving vent to anger, the second ring canal of giving vent to anger, connect the inside intercommunication of trachea, first ring canal of giving vent to anger, the second ring canal of giving vent to anger, the connecting trachea is gone up to open and.

Further, the discharge end include the discharge gate, the center department of discharge gate opens there is the standing groove, the second motor is located the center department of standing groove, the discharge gate with the inside intercommunication in reaction bin, the inside of discharge gate is equipped with the valve.

Further, the bioreactor works as follows:

adding a culture solution and hybridoma cells into a reaction bin from a feed inlet, starting a first motor and a second motor, driving a first rotating shaft to rotate by an output shaft of the first motor, driving a defoaming plate and a stirring plate to rotate by the first rotating shaft, stirring the culture solution by the stirring plate, driving a second rotating shaft to rotate by an output shaft of the second motor, driving a first main transmission gear and a second main transmission gear to rotate by the second rotating shaft, meshing the first main transmission gear with a first auxiliary transmission gear, driving a first auxiliary transmission gear to rotate by the first main transmission gear, driving a first auxiliary transmission gear to rotate by the first auxiliary transmission gear, driving a second auxiliary transmission gear to rotate by the first connecting rod, meshing the second auxiliary transmission gear with a roller gear, driving a roller gear to rotate by the roller gear, driving a first fixed shaft to rotate by the first fixed shaft, driving a first roller shaft and a second roller shaft to rotate by the roller gear, and driving a first fixed, The drive belt cooperatees, make the defoaming piece move along the drive belt, and then drive the defoaming needle along the drive belt, the defoaming needle uses first pivot to rotate as the axle center simultaneously, the second pivot drives second main drive gear and rotates, second main drive gear and stirring board gear engagement, second main drive gear drives stirring board gear rotation, stirring board gear drives stirring board pivot rotation, stirring board pivot drives stirring board and stirring curb plate rotation, make the stirring board rotate with the stirring curb plate with first connecting rod when using first pivot as the axle center pivoted, let in carbon dioxide into the inlet standpipe, carbon dioxide gets into first outlet ring canal and second outlet ring canal along the inlet pipe through the horizontal pipe of admitting air and connecting pipe, discharge by the venthole, after finishing hybridoma cell culture, open discharge gate internal valve, collect culture solution and hybridoma cell.

The invention has the beneficial effects that: the invention drives a second rotating shaft to rotate through the extension of an output shaft of a second motor, the second rotating shaft drives a first main transmission gear and a second main transmission gear to rotate, the first main transmission gear is meshed with a first auxiliary transmission gear, the first main transmission gear drives a first auxiliary transmission gear to rotate, the first auxiliary transmission gear drives a first connecting rod to rotate, the first connecting rod drives a second auxiliary transmission gear to rotate, the second auxiliary transmission gear is meshed with a roller shaft gear, the second auxiliary transmission gear drives a roller shaft gear to rotate, the roller shaft gear drives a first fixed shaft to rotate, the first fixed shaft drives a first roller shaft to rotate, the first roller shaft, the second roller shaft and a transmission belt are matched, so that a defoaming block moves along the transmission belt, a defoaming needle is further driven to move along the transmission belt, the defoaming needle rotates by taking the first rotating shaft as an axis, and bubbles generated when the defoaming needle can well cultivate hybridoma cells are eliminated, effectively avoids the culture solution from escaping, prevents the culture solution from escaping from the air outlet pipe or the shaft seal to increase the opportunity of contamination, ensures the normal culture of cells, the second main transmission gear is driven to rotate by the second rotating shaft and is meshed with the stirring plate gear, the second main transmission gear drives the stirring plate gear to rotate, the stirring plate gear drives the stirring plate rotating shaft to rotate, the stirring plate rotating shaft drives the stirring plate and the stirring side plate to rotate, so that the stirring plate rotates by taking the first rotating shaft as the axis and the stirring side plate rotates by the first connecting rod, the generation of swirling flow in the stirring process is effectively prevented, hybridoma cells are uniformly dispersed, meanwhile, the service life of the device is prolonged, the required gas is uniformly dispersed in the reaction bin through the gas inlet mechanism, so that the dissolved oxygen in each area in the reaction bin is the same, and the preparation efficiency of the monoclonal antibody is improved.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a schematic diagram of a bioreactor for monoclonal antibody production according to the present invention;

FIG. 2 is a top view of a bioreactor for monoclonal antibody production according to the present invention;

FIG. 3 is a bottom view of the discharge end of a bioreactor for monoclonal antibody production according to the present invention;

FIG. 4 is a schematic diagram of the structure A in FIG. 1;

FIG. 5 is a schematic view of the structure B in FIG. 4;

FIG. 6 is a bottom view of a bubble plate in a bioreactor for monoclonal antibody production according to the present invention;

FIG. 7 is a schematic view of structure C of FIG. 1;

FIG. 8 is a side view of a stirring plate in a bioreactor for monoclonal antibody production according to the present invention;

FIG. 9 is a schematic structural view of a gas inlet mechanism in a bioreactor for monoclonal antibody production according to the present invention;

FIG. 10 is a top view of a gas inlet mechanism in a bioreactor for monoclonal antibody production according to the present invention.

In the figure: 1. a tank body; 11. a support pillar; 12. a base; 13. a first motor; 131. a first rotating shaft; 132. a stirring plate; 1321. a stirring plate rotating shaft; 1322. a stirring plate gear; 133. stirring the side plate; 14. a second motor; 141. a second rotating shaft; 1411. a first main drive gear; 1412. a first secondary drive gear; 1413. a first link; 1414. a second secondary drive gear; 1415. a second main drive gear; 15. a heater; 151. a temperature sensor; 16. a feed inlet; 17. a discharge end; 171. a discharge port; 172. a placement groove; 18. a pH controller; 2. a reaction bin; 3. a cooling bin; 31. a water inlet pipe; 32. a water outlet pipe; 4. a defoaming plate; 41. a first roller shaft; 411. a first fixed shaft; 412. a roll shaft gear; 42. a second roller shaft; 421. a second fixed shaft; 43. a transmission belt; 431. defoaming blocks; 432. a defoaming needle; 5. an air intake mechanism; 51. an air inlet standpipe; 52. an intake cross pipe; 53. a first gas outlet ring pipe; 531. an air outlet; 54. a second gas outlet ring pipe; 55. connecting an air pipe; 6. and an air outlet pipe.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1-10, the present invention is a bioreactor for monoclonal antibody preparation, comprising a tank 1, a reaction chamber 2 disposed inside the tank 1, a first motor 13 fixed at the center of the top end of the tank 1, a first rotating shaft 131 fixed on the output shaft of the first motor 13, the first rotating shaft 131 sequentially passing through the side walls of the tank 1 and the reaction chamber 2, the first rotating shaft 131 rotatably connected with the side walls of the tank 1 and the reaction chamber 2, the bottom end of the first rotating shaft 131 rotatably connected with the inner wall of the reaction chamber 2, a plurality of oppositely disposed stirring plates 132 disposed at the two sides of the first rotating shaft 131, a second motor 14 fixed at the center of the bottom end of the tank 1, a second rotating shaft 141 fixed on the output shaft of the second motor 14, the second rotating shaft 141 sequentially passing through the side walls of the tank 1 and the reaction chamber 2, the second rotating shaft 141 rotatably connected with the side walls of the tank 1 and the reaction chamber 2, the second rotating shaft 141 is positioned inside the first rotating shaft 131, the second rotating shaft 141 is rotatably connected with the first rotating shaft 131, a plurality of second main transmission gears 1415 which are uniformly distributed are fixed on the second rotating shaft 141, a first main transmission gear 1411 is also fixed on the second rotating shaft 141, the first main transmission gear 1411 is positioned above the second main transmission gears 1415, a heater 15 is fixed on one side of the outside of the tank body 1, a temperature sensor 151 and a pH controller 18 are fixed on the side wall of the reaction bin 2, the heater 15, the temperature sensor 151 and the pH controller 18 are controlled by a PLC, a feed inlet 16 is arranged at the top end of the tank body 1, the feed inlet 16 is communicated with the inside of the reaction bin 2, a valve is arranged inside the feed inlet 16, a discharge end 17 is arranged at the center of the bottom end of the outside of the tank body 1, two defoaming plates 4 which are oppositely arranged are fixed on the first rotating shaft 131, the defoaming plates 4 are positioned above the stirring plates 132, and a plurality, defoaming piece 431's lower fixed surface has defoaming needle 432, the outside of reaction bin 2 is equipped with air inlet mechanism 5, air inlet mechanism 5 is located between reaction bin 2 and the jar body 1, air inlet mechanism 5 is including air inlet standpipe 51, the lateral wall of the jar body 1 is passed on air inlet standpipe 51's top, air inlet standpipe 51 and the lateral wall fixed connection of the jar body 1, air inlet standpipe 51's one side that is close to reaction bin 2 is fixed with a plurality of horizontal pipes 52 of admitting air, be fixed with first ring canal 53 of giving vent to anger on the horizontal pipe 52 of admitting air, first ring canal 53 of giving vent to anger is located the inside of reaction bin 2, the top of reaction bin 2 is equipped with outlet duct 6, outlet duct 6 and the inside intercommunication of reaction bin 2, outlet duct 6 passes the.

The outer bottom end of the tank body 1 is fixed with four uniformly distributed support columns 11, the bottom end of each support column 11 is fixed with a base 12, the outer side wall of the tank body 1 is fixed with a cooling bin 3, the top end of one side of the cooling bin 3 is provided with a water inlet pipe 31, the other side of the cooling bin 3 is provided with a water outlet pipe 32, the water inlet pipe 31 and the water outlet pipe 32 are respectively communicated with the inside of the cooling bin 3, and the first rotating shaft 131 and the second rotating shaft 141 rotate in opposite directions.

As shown in fig. 1, 7, and 8, the stirring plate 132 is located inside the reaction chamber 2, two stirring side plates 133 are fixed on a side wall of the stirring plate 132, the stirring side plates 133 are arc plates, a stirring plate rotating shaft 1321 is fixed at one end of the stirring plate 132 close to the first rotating shaft 131, an end of the stirring plate rotating shaft 1321 is located inside the first rotating shaft 131, the first rotating shaft 131 is rotatably connected with the stirring plate rotating shaft 1321, a stirring plate gear 1322 is fixed on an end of the stirring plate rotating shaft 1321, and the stirring plate gear 1322 is engaged with the second main transmission gear 1415.

As shown in fig. 4-6, a first fixed shaft 411 is disposed at one end of the lower surface of the defoaming plate 4 close to the first rotating shaft 131, the upper end of the first fixed shaft 411 passes through the lower surface of the defoaming plate 4, the first fixed shaft 411 is rotatably connected with the defoaming plate 4, a roller shaft gear 412 is fixed at the upper end of the first fixed shaft 411, a second sub-transmission gear 1414 is vertically disposed at one side of the roller shaft gear 412, the second sub-transmission gear 1414 is engaged with the roller shaft gear 412, a first link 1413 is fixed on the sidewall of the second sub-transmission gear 1414, a first sub-transmission gear 1412 is fixed at the end of the first link 1413, the first sub-transmission gear 1412 is engaged with the first main transmission gear 1411, a first roller shaft 41 is fixed at the lower end of the first fixed shaft 411, a second fixed shaft 421 is fixed at one end of the lower surface of the defoaming plate 4 far from the first rotating shaft 131, a second roller shaft 42 is disposed, a transmission belt 43 is arranged between the first roller shaft 41 and the second roller shaft 42, a plurality of defoaming blocks 431 which are uniformly distributed are fixed on the surface of the transmission belt 43, and defoaming needles 432 are fixed on the lower surfaces of the defoaming blocks 431.

As shown in fig. 8 and 9, a second air outlet ring pipe 54 is arranged in an inner ring of the first air outlet ring pipe 53, two opposite connecting air pipes 55 are fixed on a side wall of the second air outlet ring pipe 54, an end of the connecting air pipe 55 is fixedly connected with the first air outlet ring pipe 53, the second air outlet ring pipe 54 and the connecting air pipe 55 are communicated with each other, a plurality of air outlet holes 531 are formed in the first air outlet ring pipe 53, the second air outlet ring pipe 54 and the connecting air pipe 55, and the air inlet transverse pipes 52 and the stirring plates 132 are arranged in a staggered manner.

As shown in fig. 1 and 3, the discharging end 17 includes a discharging port 171, a placement groove 172 is formed in the center of the discharging port 171, the second motor 14 is located in the center of the placement groove 172, the discharging port 171 is communicated with the interior of the reaction chamber 2, and a valve is disposed in the discharging port 171.

Referring to FIGS. 1-10, the bioreactor for monoclonal antibody production works as follows:

adding culture solution and hybridoma cells into a reaction bin 2 from a feed inlet 16, starting a first motor 13 and a second motor 14, driving a first rotating shaft 131 to rotate by an output shaft of the first motor 13, driving a defoaming plate 4 and a stirring plate 132 to rotate by the first rotating shaft 131, stirring the culture solution by the stirring plate 132, driving a second rotating shaft 141 to rotate by an output shaft of the second motor 14, driving a first main transmission gear 1411 and a second main transmission gear 1415 to rotate by the second rotating shaft 141, driving the first main transmission gear 1411 to be meshed with a first secondary transmission gear 1412, driving a first secondary transmission gear 1412 to rotate by the first main transmission gear 1411, driving a second secondary transmission gear 1412 to rotate by the first secondary transmission gear 1412, driving a second secondary transmission gear 1414 to rotate by the first connecting rod 1413, driving the second secondary transmission gear 1412 to be meshed with a roller gear 412, driving the roller gear 412 to rotate by the roller gear 412, driving a first fixed shaft 411 to rotate by the roller gear 412, the first fixing shaft 411 drives the first roller shaft 41 to rotate, the first roller shaft 41, the second roller shaft 42 and the transmission belt 43 are matched, so that the defoaming block 431 moves along the transmission belt 43, the defoaming needle 432 is further driven to move along the transmission belt 43, meanwhile, the defoaming needle 432 rotates by taking the first rotating shaft 131 as an axis, bubbles generated when hybridoma cells are cultured can be well eliminated by the defoaming needle 432, the phenomenon that a large amount of culture solution escapes when too much foam is generated is effectively prevented, the culture solution escapes from the air outlet pipe 6 or the shaft seal to increase the chance of contamination and the like, aeration stirring cannot be carried out even in serious cases, cell respiration is blocked to cause metabolic abnormality, the second rotating shaft 141 drives the second main transmission gear 1415 to rotate, the second main transmission gear 1415 is meshed with the stirring plate gear 1322, the stirring plate gear 1322 is driven to rotate by the stirring plate gear 1322, the stirring plate gear 1322 drives the stirring plate rotating shaft 1321 to rotate, the stirring plate rotating shaft 1321 drives the stirring plate 132 and the stirring side plate 133 to rotate, so that the stirring plate 132 rotates by taking the first rotating shaft 131 as an axis and the stirring side plate 133 rotates by taking the first connecting rod 1413, the generation of eddy current in the stirring process is effectively prevented, the eddy current can enable the axial circulation speed to be lower than the radial circulation speed, the uniform dispersion of hybridoma cells is influenced, the violently swirling culture solution can cause waves which are rushed back and forth, the impact force can be generated on the equipment by combining the effect of the eddy, the service life of the equipment is further shortened, the damage to cells is caused, the preparation of monoclonal antibody is further influenced, carbon dioxide is introduced into the air inlet vertical pipe 51, enters the first air outlet circular pipe 53 and the second air outlet circular pipe 54 along the air inlet vertical pipe 51 through the air inlet horizontal pipe 52 and the connecting air pipe 55 and is discharged from the air outlet hole 531, the first air outlet circular pipe 53 and the second air outlet circular pipe 54 enable the carbon dioxide to be uniformly distributed, the uniform distribution of carbon dioxide can make the hybridoma cell grow normally, has reduced the production of unusual hybridoma cell, and temperature sensor 151 and pH controller 18 can detect the temperature and the pH value in reaction bin 2, and then keep the culture environment of hybridoma cell, and after the completion of hybridoma cell culture, open discharge gate 171 internal valve, collect culture solution and hybridoma cell.

The foregoing is merely exemplary and illustrative of the principles of the present invention and various modifications, additions and substitutions of the specific embodiments described herein may be made by those skilled in the art without departing from the principles of the present invention or exceeding the scope of the claims set forth herein.

Claims

1. A bioreactor for monoclonal antibody production, comprising: the bioreactor is used for large-scale culture of hybridoma cells in monoclonal antibody preparation, and comprises a tank body (1), wherein a reaction bin (2) is arranged inside the tank body (1), a first motor (13) is fixed at the center of the top end of the outside of the tank body (1), a first rotating shaft (131) is fixed on an output shaft of the first motor (13) in an extending manner, the first rotating shaft (131) sequentially penetrates through the side walls of the tank body (1) and the reaction bin (2), the first rotating shaft (131) is rotationally connected with the side walls of the tank body (1) and the reaction bin (2), the bottom end of the first rotating shaft (131) is rotationally connected with the inner wall of the reaction bin (2), a plurality of stirring plates (132) which are oppositely arranged are arranged on two sides of the first rotating shaft (131), a second motor (14) is fixed at the center of the bottom end of the outside of the tank body (1), a second rotating shaft (141) is fixed on an output shaft of the second motor (14), and the second rotating shaft (141) sequentially penetrates through the side walls of, the second rotating shaft (141) is rotatably connected with the side walls of the tank body (1) and the reaction bin (2), the second rotating shaft (141) is positioned inside the first rotating shaft (131), the second rotating shaft (141) is rotatably connected with the first rotating shaft (131), a plurality of second main transmission gears (1415) which are uniformly distributed are fixed on the second rotating shaft (141), a first main transmission gear (1411) is also fixed on the second rotating shaft (141), the first main transmission gear (1411) is positioned above the second main transmission gear (1415), a heater (15) is fixed on one side of the outer part of the tank body (1), a temperature sensor (151) and a pH controller (18) are fixed on the side wall of the reaction bin (2), the heater (15), the temperature sensor (151) and the pH controller (18) are controlled by a PLC, a feeding hole (16) is formed in the top end of the tank body (1), and the feeding hole (16) is communicated with the inside of the reaction bin (2), a valve is arranged in the feed inlet (16), a discharge end (17) is arranged at the center of the bottom end of the outer part of the tank body (1), two defoaming plates (4) which are oppositely arranged are fixed on the first rotating shaft (131), the defoaming plates (4) are positioned above the stirring plate (132), a plurality of defoaming blocks (431) are arranged on the lower surfaces of the defoaming plates (4), defoaming needles (432) are fixed on the lower surfaces of the defoaming blocks (431), an air inlet mechanism (5) is arranged outside the reaction bin (2), the air inlet mechanism (5) is positioned between the reaction bin (2) and the tank body (1), the air inlet mechanism (5) comprises an air inlet vertical pipe (51), the top end of the air inlet vertical pipe (51) penetrates through the side wall of the tank body (1), the air inlet vertical pipe (51) is fixedly connected with the side wall of the tank body (1), a plurality of air inlet horizontal pipes (52) are fixed on one side of the air inlet vertical pipe (51) close to the reaction bin (2), and a first air, first ring canal of giving vent to anger (53) are located the inside in reaction storehouse (2), and the top in reaction storehouse (2) is equipped with outlet duct (6), and outlet duct (6) and reaction storehouse (2) inside intercommunication, the roof of jar body (1) is passed in outlet duct (6).

2. The bioreactor for monoclonal antibody production according to claim 1, wherein: the outside bottom mounting of the jar body (1) have four evenly distributed's support column (11), the bottom mounting of support column (11) has base (12), be fixed with cooling bin (3) on the outside lateral wall of the jar body (1), one side top of cooling bin (3) is equipped with inlet tube (31), the opposite side of cooling bin (3) is equipped with outlet pipe (32), inlet tube (31) and outlet pipe (32) respectively with cooling bin (3) inside intercommunication, first pivot (131) and second pivot (141) opposite direction rotate.

3. The bioreactor for monoclonal antibody production according to claim 1, wherein: stirring board (132) be located the inside in reaction storehouse (2), be fixed with two stirring curb plate (133) that the opposition set up on the lateral wall of stirring board (132), stirring curb plate (133) are the arc board, the one end that stirring board (132) are close to first pivot (131) is fixed with stirring board pivot (1321), the end of stirring board pivot (1321) is located the inside of first pivot (131), first pivot (131) rotate with stirring board pivot (1321) and are connected, be fixed with stirring board gear (1322) on the end of stirring board pivot (1321), stirring board gear (1322) and second main drive gear (1415) mesh mutually.

4. The bioreactor for monoclonal antibody production according to claim 1, wherein: a first fixed shaft (411) is arranged at one end, close to the first rotating shaft (131), of the lower surface of the defoaming plate (4), the upper end of the first fixed shaft (411) penetrates through the lower surface of the defoaming plate (4), the first fixed shaft (411) is rotatably connected with the defoaming plate (4), a roller shaft gear (412) is fixed at the upper end of the first fixed shaft (411), a second secondary transmission gear (1414) is vertically arranged on one side of the roller shaft gear (412), the second secondary transmission gear (1414) is meshed with the roller shaft gear (412), a first connecting rod (1413) is fixed on the side wall of the second secondary transmission gear (1414), a first secondary transmission gear (1412) is fixed at the end of the first connecting rod (1413), the first secondary transmission gear (1412) is meshed with the first primary transmission gear (1411), a first roller shaft (41) is fixed at the lower end of the first fixed shaft (411), a second fixed shaft (421) is fixed at one end, far away from the first rotating shaft (131, the bottom of second fixed axle (421) is equipped with second roller (42), and second roller (42) rotate with second fixed axle (421) and are connected, installs between first roller (41) and second roller (42) drive belt (43), and the fixed surface of drive belt (43) has a plurality of evenly distributed's defoaming piece (431), and the lower fixed surface of defoaming piece (431) has defoaming needle (432).

5. The bioreactor for monoclonal antibody production according to claim 1, wherein: the inner ring of the first air outlet ring pipe (53) is internally provided with a second air outlet ring pipe (54), the side wall of the second air outlet ring pipe (54) is fixed with two oppositely arranged connecting air pipes (55), the end of each connecting air pipe (55) is fixedly connected with the first air outlet ring pipe (53), the second air outlet ring pipe (54) and the connecting air pipes (55) are communicated with each other, the first air outlet ring pipe (53), the second air outlet ring pipe (54) and the connecting air pipes (55) are provided with a plurality of air outlet holes (531), and the air inlet transverse pipes (52) and the stirring plates (132) are arranged in a staggered mode.

6. The bioreactor for monoclonal antibody production according to claim 1, wherein: discharge end (17) including discharge gate (171), the center department of discharge gate (171) opens and has standing groove (172), second motor (14) are located the center department of standing groove (172), discharge gate (171) and reaction storehouse (2) inside intercommunication, the inside of discharge gate (171) is equipped with the valve.

7. The bioreactor for monoclonal antibody production according to claim 1, wherein: the bioreactor works as follows:

adding a culture solution and hybridoma cells into a reaction bin (2) from a feed inlet (16), starting a first motor (13) and a second motor (14), driving a first rotating shaft (131) to rotate by an output shaft of the first motor (13), driving a defoaming plate (4) and a stirring plate (132) to rotate by the first rotating shaft (131), stirring the culture solution by the stirring plate (132), driving a second rotating shaft (141) to rotate by an output shaft of the second motor (14), driving a first main transmission gear (1411) and a second main transmission gear (1415) to rotate by the second rotating shaft (141), meshing the first main transmission gear (1411) with a first auxiliary transmission gear (1412), driving a first auxiliary transmission gear (1412) to rotate, driving a first connecting rod (1413) to rotate by the first auxiliary transmission gear (1412), driving a second auxiliary transmission gear (1414) to rotate by the first connecting rod (1413), the second auxiliary transmission gear (1414) is meshed with the roller shaft gear (412), the second auxiliary transmission gear (1414) drives the roller shaft gear (412) to rotate, the roller shaft gear (412) drives the first fixing shaft (411) to rotate, the first fixing shaft (411) drives the first roller shaft (41) to rotate, the first roller shaft (41), the second roller shaft (42) and the transmission belt (43) are matched, so that the defoaming block (431) moves along the transmission belt (43) and further drives the defoaming needle (432) to move along the transmission belt (43), meanwhile, the defoaming needle (432) rotates by taking the first rotating shaft (131) as an axis, the second rotating shaft (141) drives the second main transmission gear (1415) to rotate, the second main transmission gear (1415) is meshed with the stirring plate gear (1322), the second main transmission gear (1415) drives the stirring plate gear (141) to rotate, the stirring plate gear (1322) drives the stirring plate rotating shaft (1321) to rotate, stirring board pivot (1321) drive stirring board (132) and stirring curb plate (133) and rotate, make stirring board (132) use first pivot (131) as the axle center pivoted simultaneously with stirring curb plate (133) with first connecting rod (1413) rotation, let in carbon dioxide into air inlet standpipe (51), carbon dioxide gets into first outlet ring pipe (53) and second outlet ring pipe (54) along air inlet standpipe (51) through air inlet horizontal pipe (52) and connecting trachea (55), discharge by venthole (531), after hybridoma cell culture, open discharge gate (171) internal valve, collect culture solution and hybridoma cell.