CN100375782C - Bioreactor system for animal large scale cell culture - Google Patents
Bioreactor system for animal large scale cell culture Download PDFInfo
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- CN100375782C CN100375782C CNB2005100301974A CN200510030197A CN100375782C CN 100375782 C CN100375782 C CN 100375782C CN B2005100301974 A CNB2005100301974 A CN B2005100301974A CN 200510030197 A CN200510030197 A CN 200510030197A CN 100375782 C CN100375782 C CN 100375782C
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
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Abstract
The present invention discloses a biological reactor system for large-scale animal cell culture, which comprises a reactor provided with a fully closed magnetic transmission device, a mixing chamber connected with the reactor by a pipe line and a gas supply system connected with the mixing chamber by the pipe line. The fully closed magnetic transmission device comprises an upper bearing, a lower bearing, an inner magnetic transmission shaft, a combined oar blade assembly, a non-magnetic sealing sleeve, outer magnetic steel and a lead-magnetic cylinder, wherein the outer magnetic steel is separated from the inner magnetic transmission shaft by the fully closed non-magnetic sealing sleeve. A channel is thoroughly separated, microbes or other pollutants enter the biological reactor from a stirring system by the channel, and the opportunity of pollution because of the entering process of the microbes or the pollutants is eliminated. The present invention has the advantages of simple structure, large transmitted power and uniform material mixing effect in the reactor, and the present invention can meet the needs of the industrial development of the cell culture.
Description
Technical field
The present invention relates to a kind of bioreactor system, relate in particular to a kind of bioreactor system that is used for extensive animal cell culture.
Technical background
Since the eighties, the great achievement that zooblast genetically engineered and cell-fusion techniques are obtained makes and can express and produce all kinds of diagnosis and treat biological activity proteins such as using monoclonal antibody, vaccine, somatomedin by the zooblast vitro culture.Because the curative effect of these protein and monoclonal antibody is very clear and definite, has very wide market application prospect, therefore using these high value medicines of zooblast vitro culture technology mass production has become the integral part of biological medicine hi-tech industry, is the key areas of current new drug development and production.
Use modern bioreactor system and replace original manual bottle or the square vase culture systems of rolling, can simplify production process greatly, enhance productivity and the product homogeneity, reduce production costs, guarantee security, validity, reliability and the economy of product clinical application.Britain Celltech company uses 100～10000 liters airlift bioreactor cultivation hybridoma manufacture order clonal antibody; The bio-reactor scale that Japan Sumitomo company is used to produce tPA reaches 8000 liters; Wellcome company uses stirred bioreactor production Interferon, rabbit and other biological products of 8000 liters; The bio-reactor scale that Germany Behringwerke company is used to produce erythropoietin (rhEPO) reaches 15m
3Germany Bayer company adopts the high-density continous pouring culture technique to produce blood coagulation factor VIII in the subsidiary factory of Los Angeles,U.S, and the continuous production cycle reaches three to six months, becomes the longest biological product of continuous production cycle of drugs approved by FDA up to now.
Yet, the domestic production technology adopts tradition to roll the flask culture method mostly at present, so-called suitability for industrialized production also just adopts 5 liters of CelliGen Plus fixed-bed reactor of U.S. NBS company, industrial scale is little, be difficult to amplify, and production process is difficult to further optimization, causes output little, production efficiency is low, and is far apart with actual needs.Therefore, the research and development of animal cell large-scale high-density serum-free culture technology and bio-reactor thereof is used, and has become the important factor of restriction China biotech development, presses for to be solved.
For bio-reactor, it is to prevent microbiological contamination that the especially secular continous pouring of the large scale culturing of zooblast is cultivated what at first need to solve.Its envelope of prior biological reactor stirring system mostly adopts the mode of mechanical seal, its advantage is that power of agitator is big, but often because the material that is adopted, the reason of design and working accuracy aspect, system's stopping property in use is not enough, thereby especially in secular culturing process, can produce to leak and cause microbiological contamination, in order to address this problem, more external in recent years companies begin to adopt the magnetic agitation system, its shaft seal structure of this stirring system is relatively complicated, and because the principle that adopts magnetic force to transmit, cause its power of agitator to reduce greatly, be mainly used at present in the meiofauna biological reactor for cell culture.
Zooblast will be from original square vase, roll flask culture is amplified to the bioreactor culture of various scales and realizes high-density culture process, faces the amplification and the reinforcement problem of culturing process and bio-reactor with regard to having to.Amplification and reinforcement problem are mainly derived from the contradiction of two aspects: on the one hand, the bio-reactor that is used for culturing process must have enough gas-liquid mass transfer abilities and good fluid mixing condition can be provided, otherwise just be difficult to the sustenticular cell growth, especially this requirement will be harsh more in the large scale and high density cell cultivation process; On the other hand, because the shearing sensibility of zooblast, because of stir and fluid shearing that ventilation produces and bubble phenomenon can cause cell injury even breakage, death unavoidablely, make the culturing process failure in the bio-reactor.Therefore, the design of stirring arm is a very important problem in the animal cell culture bioreactor, both must guarantee that the mass transfer that provides good mixed, and the fluidic shearing force is reduced to reduce the damage of pair cell.
PH and dissolved oxygen be by air, oxygen, the related control of four kinds of gases of nitrogen and carbonic acid gas in the reactor, and it is that the Controlling System of reactor realizes by controlling four kinds of gases magnetic valve switching time separately under the PID pattern that this association is controlled.But in actually operating, enter its pressure and unbalanced separately of gas of reactor by steel cylinder and air compressor, this unbalanced meeting causes the higher excessive feeding of gas of a certain or several pressure, the gas that other pressure is lower then can't enter reactor, thereby the control of pH and dissolved oxygen is occurred than large deviation; In addition, the unbalanced all gases flux error that causes of this pressure can't be corrected by the PID master mode, and consequently pH and dissolved oxygen produce violent spination fluctuation.
Summary of the invention
The technical issues that need to address of the present invention are to disclose a kind of bioreactor system that is used for extensive suspension cell culture, to overcome the above-mentioned defective that prior art exists, satisfy the needs of cell cultures industrialized development.
Bioreactor system of the present invention comprises:
Be provided with the reactor of totally enclosed magnetic drive device;
The mixing section that is connected by pipeline with reactor;
The airing system that is connected by pipeline with mixing section;
Said totally enclosed magnetic drive device comprises head bearing, lower bearing, interior magnetic transmission shaft, built-up type paddle assembly, no magnetic seal cover, outer steel and magnetic conductive cylinder;
Said outer steel is arranged on that no magnetic seal cover is outer, between head bearing and the lower bearing; Motor by the outside drives, as the propulsion source of whole stirring system;
The bottom of said no magnetic seal cover is connected with the top seal of reactor, no magnetic seal cover is a fixed, totally enclosed sleeve, outer and the reactor exterior of no magnetic seal cover, communicate with inside reactor in the no magnetic seal cover, it with outer steel, interior magnetic transmission shaft all less than being connected, play inside and outside isolation action;
Corresponding with outer steel with interior magnet steel on the magnetic transmission shaft in said, interior magnetic transmission shaft is not had the magnetic seal cover and is fixed at interval and by neck bearing.The lower end of interior magnetic transmission shaft is a round bar, and round bar passes reactor head, stretches in the reactor and with built-up type paddle assembly to be connected;
It is outer but be not connected with outer steel that said magnetic conductive cylinder is wrapped in outer steel, shields;
Said built-up type paddle assembly comprises stir shaft and the three layers of paddle that are connected on the interior magnetic transmission shaft, two-layer paddle is oblique leaf hoisting type paddle down, make fluid produce axial flow, reduce the hydrodynamic shear that stirring blade causes greatly, the upper strata paddle is curved leaf turbine slurry, can make fluid produce radial flow, strengthen mass transfer and mixing in the reactor.
Reactor of the present invention is operation like this:
The electric motor driving outer steel drives interior magnetic transmission shaft thereby rely on magnetic force to drive interior magnet steel, thereby drives the rotation of built-up type paddle assembly.No magnetic seal cover and magnetic conductive cylinder are mounting blocks, wherein do not have magnetic seal and have overlapped inside and outside isolation action, and magnetic conductive cylinder is used for protecting outer steel.
By above-mentioned disclosed technical scheme as seen, separate with totally enclosed no magnetic seal cover between outer steel and the interior magnetic transmission shaft, thoroughly completely cut off microorganism or other pollutent are entered bio-reactor by stirring system passage, eliminated the opportunities for contamination that causes therefrom, the present invention is simple in structure, transmitted power is big, and the mixing of materials in the reactor is even, can satisfy the needs of cell cultures industrialized development.
Description of drawings
Fig. 1 is the structural representation of bioreactor system.
Fig. 2 is totally enclosed magnetic drive device structural representation.
Fig. 3 is a built-up type paddle assembly.
Fig. 4 is curved leaf turbine slurry vertical view.
Fig. 5 is the airing system synoptic diagram.
Embodiment
Referring to Fig. 1, bioreactor system of the present invention comprises:
The totally enclosed magnetic drive device 101 that is used for transmission reactor stir shaft;
The mixing section 102 that is connected by pipeline with reactor;
The airing system 103 that is connected by pipeline with mixing section 102;
Referring to Fig. 2, said totally enclosed magnetic drive device comprises head bearing (1), lower bearing (6), interior magnetic transmission shaft (2), no magnetic seal cover (3), outer steel (4) and magnetic conductive cylinder (5), neck bearing (7), built-up type paddle assembly;
Said outer steel (4) is arranged on that no magnetic seal cover (3) is outer, between head bearing (1) and the lower bearing (6); Motor by the outside drives, as the propulsion source of whole stirring system;
The top cover (203) of the bottom of said no magnetic seal cover (3) and reactor is tightly connected, and no magnetic seal cover (3) is a fixed, totally enclosed sleeve, it with outer steel (4), interior magnetic transmission shaft (2) all less than being connected, play inside and outside isolation action;
Magnetic transmission shaft (2) is upward corresponding with outer steel with interior magnet steel (201) in said, and interior magnetic transmission shaft (2) is not had magnetic seal cover (3) interval and is fixed by neck bearing (7).The lower end of interior magnetic transmission shaft (2) is round bar (202), and round bar (202) passes reactor head (203), stretches in the reactor and with built-up type paddle assembly (8) to be connected;
Said magnetic conductive cylinder (5) is wrapped in outside the outer steel (4), shields;
Referring to Fig. 3 and Fig. 4, said built-up type paddle assembly comprises stir shaft and the three layers of paddle that are connected on the interior magnetic transmission shaft (2), two-layer paddle (501) is oblique leaf hoisting type paddle down, upper strata paddle (502) is curved leaf turbine slurry, tiltedly the inclined-plane and the angle [alpha] between the horizontal plane of leaf hoisting type paddle are 20 °-40 °, and the arc radius of curved leaf turbine slurry paddle (502) is 250mm.
Referring to Fig. 5, said airing system 103 comprises that an end is connected with carbon dioxide air source with air source of the gas, oxygen source, nitrogen source of the gas respectively, the pipeline of the other end and mixing section 102, pipeline is provided with magnetic valve 601 and non-return valve 602, the effect of non-return valve is to prevent the excessive feeding of gas that a certain or several pressure are higher under four kinds of unbalanced situations of gaseous tension, and makes the lower gas of other pressure can't enter reactor; Gas mixing chamber has then played the buffered effect, has avoided the spination fluctuation of interior pH of reactor and dissolved oxygen.
Claims (3)
1. bioreactor system that is used for the animal large scale cell cultures comprises:
The totally enclosed magnetic drive device (101) that is used for transmission reactor stir shaft;
The gas mixing chamber (102) that is connected by pipeline with reactor;
The airing system (103) that is connected by pipeline with mixing section (102);
It is characterized in that said totally enclosed magnetic drive device comprises head bearing (1), lower bearing (6), interior magnetic transmission shaft (2), no magnetic seal cover (3), outer steel (4) and magnetic conductive cylinder (5), neck bearing (7), built-up type paddle assembly;
Said outer steel (4) is arranged on that no magnetic seal cover (3) is outer, between head bearing (1) and the lower bearing (6); Motor by the outside drives, as the propulsion source of whole stirring system;
The top cover (203) of the bottom of said no magnetic seal cover (3) and reactor is tightly connected, and no magnetic seal cover (3) is a fixed, totally enclosed sleeve, it with outer steel (4), interior magnetic transmission shaft (2) all less than being connected, play inside and outside isolation action;
Magnetic transmission shaft (2) is gone up with interior magnet steel (201) corresponding with outer steel in said, interior magnetic transmission shaft (2) is not had magnetic seal cover (3) and also is fixed by neck bearing (7) at interval, the lower end of interior magnetic transmission shaft (2) is round bar (202), round bar (202) passes reactor head (203), stretches in the reactor and with built-up type paddle assembly to be connected;
Said magnetic conductive cylinder (5) is wrapped in outside the outer steel (4), shields;
Said built-up type paddle assembly comprises stir shaft and the three layers of paddle that are connected on the interior magnetic transmission shaft (2), and following two-layer paddle (501) is oblique leaf hoisting type paddle, and upper strata paddle (502) is curved leaf turbine slurry;
Said airing system (103) comprises the pipeline that an end is connected with carbon dioxide air source with air source of the gas, oxygen source, nitrogen source of the gas respectively, the other end is provided with is connected with mixing section (102), and pipeline is provided with magnetic valve (601) and non-return valve (602).
2. the bioreactor system that is used for the animal large scale cell cultures according to claim 1 is characterized in that, tiltedly the inclined-plane and the angle [alpha] between the horizontal plane of leaf hoisting type paddle are 20 °-40 °.
3. the bioreactor system that is used for the animal large scale cell cultures according to claim 2 is characterized in that, the arc radius of curved leaf turbine slurry paddle (502) is 250mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100301974A CN100375782C (en) | 2005-09-29 | 2005-09-29 | Bioreactor system for animal large scale cell culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100301974A CN100375782C (en) | 2005-09-29 | 2005-09-29 | Bioreactor system for animal large scale cell culture |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1766080A CN1766080A (en) | 2006-05-03 |
| CN100375782C true CN100375782C (en) | 2008-03-19 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100301974A Expired - Fee Related CN100375782C (en) | 2005-09-29 | 2005-09-29 | Bioreactor system for animal large scale cell culture |
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| CN (1) | CN100375782C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101752988B (en) * | 2008-12-16 | 2011-11-23 | 吴上能 | Magnetic transmission structure of water flow spraying device |
| KR20120031083A (en) * | 2009-07-24 | 2012-03-29 | 에프. 호프만-라 로슈 아게 | Stirrer system |
| CN103667056A (en) * | 2013-12-26 | 2014-03-26 | 南京比瑞生物科技有限公司 | Candle type bioreactor |
-
2005
- 2005-09-29 CN CNB2005100301974A patent/CN100375782C/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 动物细胞培养用生物反应器设计原理. 谭文松，戴干策，陈因良.生物工程学报，第1996,12(2)期. 1996 * |
| 造血细胞体外悬浮培养和生物反应器开发. 迟占有，夏泉鸣，康自珍，谭文松，戴干策.生物工程学报，第2003,19(5)期. 2003 * |
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| Publication number | Publication date |
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| CN1766080A (en) | 2006-05-03 |
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