CN101985597B - Animal cell reactor with bubble-free aerating system - Google Patents
Animal cell reactor with bubble-free aerating system Download PDFInfo
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- CN101985597B CN101985597B CN 201010275887 CN201010275887A CN101985597B CN 101985597 B CN101985597 B CN 101985597B CN 201010275887 CN201010275887 CN 201010275887 CN 201010275887 A CN201010275887 A CN 201010275887A CN 101985597 B CN101985597 B CN 101985597B
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Abstract
The invention discloses an animal cell reactor with a bubble-free aerating system, which comprises a reactor tank and a tank cover, and also comprises the bubble-free aerating system, wherein the bubble-free aerating system comprises a framework, at least two aerating hose connecting pieces, an aerating hose, and at least two in-place sterilization connecting pieces, wherein the framework is arranged in the reactor tank; the at least two aerating hose connecting pieces are arranged on the framework and are provided with at least two aerating interfaces; the aerating hose is wound on the framework and is communicated between the aerating interfaces of the two aerating hose connecting pieces; and the at least two in-place sterilization connecting pieces are inserted in the reactor tank and are respectively communicated with the aerating interfaces of the aerating hose connecting pieces. The bubble-free aerating system applied to the animal cell reactor can realize in-place sterilization of the animal cell reactor with the bubble-free aerating system in pilot plant tests and large-scale production.
Description
Technical field
The present invention relates to the bioreactor technology field, particularly relate to a kind of zooblast reactor with bubble-free aerating system.
Background technology
Bio-reactor is widely used in animal and plant cell cultures (or reaction) and microbial fermentation field.Wherein, the bio-reactor that relates to zooblast has special requirement.
Since the seventies, cell cultures has very large development with bio-reactor, and kind is more and more, and scale is increasing.Kind roughly has: plastics bag multiplier (-icator), filling multiplier (-icator), many plates multiplier (-icator), spiral membrane multiplier (-icator), tubular type spiral multiplier (-icator), the cellular multiplier (-icator) of ceramic rectangular channel, fluidized-bed reactor, tubular fibre and other film reactors, stirred tank reactor, air-lift reactor etc.The stirring pattern of stirred tank reactor has haply: the double-deck cage ventilating agitator of paddle stirrer, bar-shaped agitator, ship propulsion paddle stirrer, inclination blade agitator, sail shape agitator, double vibrations taper hole sieve plate agitator, cage ventilating agitator and Huadong Chemical College exploitation etc.
Along with the increase of market to cell cultures product demand, the design of external many companies has also been produced many maxicells cultivation reactors.Set up the bio-reactor of 10,000L scale such as Celltech company, cultivated hybridoma manufacture order clonal antibody; Sumitomo company has set up 8,000L reactor and has produced t-PA; Wellcome company has set up 8,000L stirred reactor and has produced virus vaccines, Interferon, rabbit and other biological products; Canada buys 42 from Switzerland Bioengineering company recently, and the 500L reactor is produced poliovirus vaccine.With regard to the biological products of producing are worth, all be scale operation undoubtedly.Yet the reactor of development of new will be an important Task with the needs that adapt to production all the time.The animal cell culture process is because the difference of its anti-shearing force has the bio-reactor that mechanical stirring, microvesicle are ventilated, the animal cell culture of bubble-free aerating system combination is used.Special in the bio-reactor with bubble-free aerating system, because this class reactor can provide required appearance oxygen requirement in the zooblast growth, the shearing force of generation is less, and does not produce foam in ventilation, avoided some common in other reactors weakness, such as foam.The bio-reactor that has at present bubble-free aerating system the Biostat Type B bio-reactor of German Bei Lang is arranged, but its working volume is respectively 1L, 2L, 5L and 10L, and bubble-free aerating system can only be the sterilization of offing normal of autoclaving still with tank body.Do not use and also on larger zooblast reactor at present, when greatest problem was sterilization in place, the sterilisation vapour pressure that sebific duct can't bear steam during by sebific duct broke.
Summary of the invention
The object of the present invention is to provide a kind of zooblast reactor with bubble-free aerating system, to solve the problem of existing zooblast reactor sterilization in place.
Technical scheme of the present invention:
A kind of zooblast reactor with bubble-free aerating system comprises reactor tank body and cover, also comprises bubble-free aerating system, and this bubble-free aerating system comprises: skeleton, be located in the described reactor tank body; At least two steam hose web members are located on the described skeleton, and this steam hose web member has at least two ventilation interfaces; Steam hose is wound on the described skeleton, and is communicated between the ventilation interface of described two steam hose web members; At least two sterilization web members in place are inserted on the described reactor tank body, and are communicated with a ventilation interface of described steam hose web member respectively.
Further, described sterilization web member in place comprises: the balance cavity; The air inlet transition piece is positioned at an end of described balance cavity, and is communicated with described balance cavity, inserting described reactor tank body, and is communicated with a ventilation interface of described steam hose web member; Inlet pipe is positioned at the other end of described balance cavity, and is communicated with described balance cavity; Guide pipe is positioned at described balance cavity, and an end is positioned at described air inlet transition piece and described balance cavity connectivity part, and and described air inlet transition piece between have a space, the other end is arranged in the sidewall of described balance cavity; Vapor pipe is positioned at the sidewall of described balance cavity, and is communicated with the other end of described guide pipe.
Further, the end of described air inlet transition piece has the shape of setting one's teeth on edge.
Further, described balance cavity comprises: sheath, and described air inlet transition piece is positioned at an end of this sheath, and is communicated with this sheath; Sleeve pipe, an end is communicated with the other end of described sheath, and the other end is communicated with described inlet pipe, has an opening on its sidewall, and the other end of described guide pipe is arranged in this opening, and described vapor pipe is arranged at this opening part, to be communicated with described guide pipe.
Further, described air inlet transition piece, sheath, sleeve pipe and inlet pipe are one-body molded.
Further, described zooblast reactor with bubble-free aerating system also comprises: locking piece, be socketed on the described sleeve pipe, and between described sheath and vapor pipe.
Further, described sheath comprises: first part, and its external diameter linear decrease, and the external diameter smallest end is connected with described air inlet transition piece; Second section, its external diameter be greater than the maximum outside diameter of first part, and be connected with first part maximum outside diameter end; Third part, its external diameter are greater than the second section external diameter, and described locking piece is between the third part and vapor pipe of described sheath.
Further, the second section end of described sheath has a groove.
Further, three of described sheath parts are formed in one.
Further, the quantity of described steam hose is 1 to 4.
Further, the ventilation interface quantity that has of described steam hose web member is 2 to 5.
Further, described skeleton is provided with fixed support, to be fixedly connected with sealing of tank cover.
Further, described fixed support is 3 or 4.
Further, the described skeleton of stating is provided with by taking part, and described reactor tank body inwall is provided with by taking part and supports, described skeleton is fixed in the described reactor tank body by taking part by described.
Further, described zooblast reactor is provided with the microvesicle blow and vent system.
Further, described zooblast reactor is provided with online cell micro observation instrument.
Further, described zooblast reactor is provided with the one or more combination in pH sensor, dissolved oxygen sensor, temperature sensor, pressure transmitter or the froth breaking sensor.
Further, described zooblast reactor is provided with tank body LOAD CELLS or feed supplement weighing system.
Further, described zooblast reactor is provided with stirring system.
Further, be provided with revolving filter in the tank body of described zooblast reactor.
In sum, more than be used for the zooblast reactor without the bubbles for aeration system, can realize the sterilization in place of the zooblast reactor with bubble-free aerating system of pilot scale and large scale of production.In addition, because bio-reactor of the present invention can be installed revolving filter, stirring system, microvesicle system, bubble-free aerating system simultaneously, it will be appreciated by those skilled in the art that each parts of the present invention and tank body type of attachment are demountable structure, thereby thereby can determine according to the demand of the animal cell culture technique of different varieties whether all or part of employing reaches the best effect of cultivating of zooblast.In addition, the use of multiple sensors of the present invention be so that can carry out correlation analysis to each parameter that reflects the cell physiological characteristic in the culturing process, thereby instructs the optimization of animal cell culture process.
Description of drawings
The structural representation with the zooblast reactor of bubble-free aerating system that Figure 1 shows that one embodiment of the invention provides.
The structural representation with the sterilization web member in place of the zooblast reactor of bubble-free aerating system that Figure 2 shows that one embodiment of the invention provides.
The part-structure synoptic diagram with the sterilization web member in place of the zooblast reactor of bubble-free aerating system that Figure 3 shows that the present invention's one implementation column provides.
The structural representation with the skeleton of the zooblast reactor of bubble-free aerating system that Figure 4 shows that the present invention's one implementation column provides.
The structural representation with a kind of steam hose web member of the zooblast reactor of bubble-free aerating system that Figure 5 shows that the present invention's one implementation column provides.
The structural representation with the another kind of steam hose web member of the zooblast reactor of bubble-free aerating system that Figure 6 shows that the present invention's one implementation column provides.
The structural representation with the zooblast reactor of bubble-free aerating system that Figure 7 shows that the present invention's one better implementation column provides.
Normal cultivation and the sterilization process schematic flow sheet in place with the zooblast reactor of bubble-free aerating system that Figure 8 shows that the present invention's one implementation column provides.
Embodiment
For technical characterictic of the present invention is become apparent, below in conjunction with accompanying drawing, provide specific embodiment, the present invention will be further described.
Please refer to Fig. 1, the structural representation with the zooblast reactor of bubble-free aerating system that it is depicted as that one embodiment of the invention provides.As shown in the figure, this zooblast reactor comprises reactor tank body 10, cover 20.Wherein, be provided with skeleton 31 in the reactor tank body 10, be wound with steam hose 32 on the skeleton 31, and be provided with at least two steam hose web members 33.Have at least two ventilation interfaces 331 (as shown in Figure 5) on this steam hose web member 33, and steam hose 32 is communicated between the ventilation interface 331 of two steam hose web members 33, particularly, after a ventilation interface 331 of one end of steam hose 32 and a steam hose web member 33 is connected, after skeleton 31 twined, the other end then was connected with a ventilation interface 331 of another steam hose web member 33.Also be inserted with at least two sterilization web members 34 in place on the reactor tank body 10, it is communicated with a ventilation interface of steam hose web member 33, for example is communicated with by silicone tube 35.
Above-described steam hose 32 is the porous steam hose, and for example the micropore emulsion tube has very small hole on it.
The below describes the principle of work of above zooblast reactor in detail take two sterilization web members 34 in place and two steam hose web members 33 as example:
When normal the cultivation, the flow route of reactant gases is as follows: air intake filter → first sterilization web member in place → first steam hose web member → steam hose → the second steam hose web member → second sterilization web member → discharge filter in place.Be that reactant gases is sent into and discharged through two sterilization web members in place, utilize the steam hose web member to send into steam hose, and the steam hose of flowing through, because steam hose has microvoid structure, reactant gases just small portion enters in the reactor tank body, and can not produce bubble in tank body, thereby realizes bubble-free aerating system, certainly, the pressure that needs control to send into reactant gases this moment is slightly larger than the gaseous tension in the reactor tank body.Particularly, the gaseous tension of keeping exactly in the steam hose 32 can not surpass bubbling pressure, and namely bubble is in the static interior pressure of film outer surface appearance.For nonwetting hydrophobic membrane, with respect to the pressure in bubbles of water about 13 * 10
2Pa, above-mentioned this requirement is to realize easily, the pressure that namely is added on the pipe should exceed about 10% than the summation of flowing pressure and pressure in bubbles in the pipe.In this case, just can form the outer liquid-gas interface layer of pipe.Even the independent bubble that moves when steam hose 32 does not occur, the situation of its mass transfer is basically similar to gas sparging.
During sterilization in place, need to carry out disinfection to whole bubble-free aerating system, when control vapour stream access line is identical with the flow route of above reactant gases, can realize the sterilization to steam hose and steam hose web member.But the sterilization of sterilization web member in place is undesirable, often has the dead angle of failing to sterilize.In the present invention's one better implementation column, provide a kind of desirable sterilization web member in place, it has avoided the appearance at dead angle in the sterilizing process in place for this reason.
Specifically, please refer to Fig. 2, this sterilization web member 34 in place comprises balance cavity 341, air inlet transition piece 342, inlet pipe 343, guide pipe 344 and vapor pipe 345.Wherein air inlet transition piece 342 is in order to insertion reaction device tank body 10, and to be communicated with a ventilation interface of steam hose web member, this air inlet transition piece 342 is positioned at an end of balance cavity 341, and is communicated with balance cavity 341; Inlet pipe 343 is positioned at the other end of balance cavity 341, and is communicated with balance cavity 341; Guide pipe 344 is positioned at the connectivity part that balance cavity 341, one ends are positioned at air inlet transition piece 342 and balance cavity 341, and and air inlet transition piece 342 between have a space, the other end is arranged in the sidewall of balance cavity 341; 345 sidewalls that are positioned at balance cavity 341 of vapor pipe, and be communicated with the other end of guide pipe.
Below in conjunction with Fig. 8, describe normal cultivation and the sterilization process flow process in place of the bubble-free aerating system of above zooblast reactor in detail.
As shown in the figure, be provided with sterilization web member 34 in place on the tank body 10 of zooblast reactor, be in the present embodiment two.Air, oxygen, carbonic acid gas, four kinds of gases of oxygen are communicated with the inlet pipe 343 of a sterilization web member 34 in place behind valve, pipeline after by air intake filter 81 by four gas mixing tanks 100 again, and vapor pipe 345 is communicated with blow-off pipe 90.The inlet pipe 343 of another sterilization web member 34 ' in place is communicated with discharge filter 82, and vapor pipe 345 ' is communicated with blow-off pipe 90.
When normal the cultivation, vapor pipe 345 and 345 ' cuts out, cultivate desired gas enters sterilization web member 34 ' in place by four gas mixing tanks 100 after air intake filter 81 filters inlet pipe 343 ', then be sent in the reactor tank body 10 via steam hose 32, because steam hose has microvoid structure, reactant gases just small portion enters in the reactor tank body, and can not produce bubble in tank body, thereby realizes bubble-free aerating system.Other gases then export to rear discharge of discharge filter 82 filtrations via the inlet pipe 343 of another one sterilization web member 34 in place.
When bio-reactor carries out disinfection, vapor pipe 345 and 345 ' is opened, the inlet pipe 343 that steam enters respectively sterilization web member 34 ' in place and 34 by steam manifold 200 through take-off pipe 2001 and 2002, and 343.Sterilization web member 34 ' in place and 34 is independently carried out sterilization in place.Further combined with Fig. 2, take sterilization web member 34 in place as example, steam enters in balance cavity 341 and the air inlet transition piece 342, owing to have certain space between guide pipe 344 and the air inlet transition piece 342, utilize this space can control flow and the pressure of steam, when vapor pressure surpasses the bubbling pressure of porous steam hose 32, steam just can be drained from the vapor pipe 345 that the space flows on balance cavity 341 sidewalls, guaranteed that porous steam hose 32 gaseous tensions can not surpass bubbling pressure in the reactor, and then finished the sterilization in place of whole bubble-free aerating system.
As seen, sterilization web member 34 in place is when carrying out sterilization in place, and the steam body in the whole sterilization web member in place 34 of can flowing through guarantees that sterilization is without the dead angle, and steam hose can't bear the problem that the pressure of steam breaks and occurs when it had avoided sterilisation vapour by steam hose.
Need to prove, more than the inlet pipe 343 of sterilization web member 34 in place can exchange with the function of vapor pipe 345, that is to say and can utilize vapor pipe 345 air inlets, inlet pipe 343 exhausts.At this moment, steam or reaction desired gas enter into guide pipe 344 via vapor pipe 345, and enter into air inlet transition piece 342 via guide pipe 344.Has certain space between guide pipe 344 and the air inlet transition piece 342, utilize this space can control flow and the pressure of steam, when vapor pressure surpasses bubbling pressure, steam just can flow to from the space inlet pipe 343 of balance cavity 341 1 ends, guaranteed that the gaseous tension in the steam hose 32 can not surpass bubbling pressure in the reactor, and then finished the sterilization in place of whole still aeration status.Its principle is constant, and the riding position that only needs to change gas filter in the whole system gets final product.Be the function that the present invention does not limit inlet pipe 343 and vapor pipe 345, they can be as air inlet or exhaust-duct.
In a preferred embodiment of the present invention, the end 3421 of air inlet transition piece 342 has the shape of setting one's teeth on edge (as shown in Figure 3), makes it tightr with being connected of silicone tube 35.Yet the present invention is as limit, and those skilled in the art can be as required arrange the end 3421 of air inlet transition piece 342.
In a preferred embodiment of the present invention, the balance cavity 341 of sterilization web member in place can assemble, also can be one-body molded.Specifically as shown in Figure 2, it provides two parts that consist of this balance cavity, is respectively sheath 3411 and sleeve pipe 3412.Wherein, air inlet transition piece 342 is positioned at an end of sheath 3411, and is communicated with this sheath 3411; One end of sleeve pipe 3412 is communicated with the other end of sheath 3411, the other end is communicated with inlet pipe 343, and has an opening on the sidewall of sleeve pipe 3412, and the other end of guide pipe 344 just is arranged in this opening, and vapor pipe 345 is arranged at this opening part, to be communicated with guide pipe 344.Better, vapor pipe 345 is welded on the sidewall of sleeve pipe 3412, and the other end of guide pipe 344 passes the opening on sleeve pipe 3412 sidewalls and is deep in the vapor pipe 345.In addition, between the sheath 3411 of this balance cavity 341 and the vapor pipe 345 locking piece 3413 can also be set, this locking piece 3413 is socketed on the sleeve pipe 3412, when being inserted on the tank body 10 with convenient sterilization web member 34 in place, it is locked on the tank body 10.
As can be seen from the figure, sheath 3411 can with air inlet transition piece 342 structure that is formed in one; Sleeve pipe 3412 and inlet pipe 343 structure that can be formed in one.This makes relevantly with die sinking, and the present invention is not as limit, and those skilled in the art can make different moulds as required, makes them separately make again assembling or one-body moldedly all can.Equally, to the composition of balance cavity 341, the present invention also is not restricted to above embodiment, those skilled in the art can be as required with its one-body molded manufacturing or offer more multi-mould and make.For example, with air inlet transition piece 342, sheath 3411, sleeve pipe 3412, inlet pipe 343 one-body molded manufacturings, then locking piece 3413 is socketed on the sleeve pipe 3412, again vapor pipe 345 is welded on the sleeve pipe 3412, to guarantee that locking piece 3413 can be from landing on the sleeve pipe 3412.Better, this locking piece 3413 is check nut, it has internal thread, when being inserted on the tank body 10 with convenient sterilization web member 34 in place, it is locked on the tank body 10.
Describe the structure of sheath 3411 in detail below in conjunction with Fig. 3, in this implementation column, sheath 3411 comprises three parts, their structures that is formed in one.The wherein external diameter linear decrease of the A of first part, and external diameter smallest end is connected with air inlet transition piece 342; The external diameter of second section B is greater than the maximum outside diameter of first part, and is connected with first part maximum outside diameter end, and this is for when sterilization web member 34 in place plugs on the bio-reactor, second section B is blocked in outside the inwall of reactor tank body; The external diameter of third part C can be located at locking piece 3413, third part C and sleeve pipe 3412 gears outside the outer wall of reactor tank body like this greater than the external diameter of second section B.Better, can offer a groove B1 at the second section B of sheath 3411 outer wall, can be at the in-built O RunddichtringO of this groove B1, when sterilization web member in place plugs the bio-reactor tank body, make between itself and the tank body by this O RunddichtringO to seal, guaranteed that the bio-reactor tank body is with extraneous isolated.
Need to prove, although above explanation is as example take two sterilization web members in place, yet the present invention is not limited in the quantity of position sterilization web member, for example, three sterilization web members in place can be set on the bio-reactor, these sterilization web members in place become 120 degree evenly to distribute, can certainly be more.So under normal cultivation conditions, need to select one of them or several as diffuser, other are as gas barrier, perhaps close one of them or several so that the selecting flexibly of different biological respinses.
Describe skeleton 31 and the structure of the steam hose on skeleton 31 32 and steam hose web member 33 and the connection between them in detail below in conjunction with Fig. 4.As can be seen from the figure, steam hose 32 is to be wrapped on the skeleton 31, usually in order to make things convenient for steam hose to twine, to utilize a plurality of support bars 311 to make cylindrical skeleton 31, better, each support bar 311 has wavy edge, and steam hose twines and is not easy thereon displacement like this.In the present embodiment, also be provided with fixed support 312 on the skeleton 31, to be sealedly and fixedly connected with cover 20.Concrete fixed support 312 bottoms have helicitic texture 3121, when mounted, this skeleton 31 is inverted in the reactor tank body 10, fixed support 312 up, the helicitic texture 3121 of fixed support 312 bottoms extend in the reservation screw on the cover 20, then, by set nut fixed support 312 is locked on the cover.Usually, fixed support 312 has 3 or 4 just can lock, and certainly, the present invention is not as limit, and those skilled in the art can select 2 or more as required.
Corresponding with this mounting means, please refer to Fig. 1, it has provided similar alternative arrangements.As shown in the figure, skeleton 31 is provided with by taking part 313, and reactor tank body 10 inwalls are provided with by taking part and support 11, skeleton 31 is fixed in the reactor tank body 10 by taking part 313 by this.The present invention does not limit the mounting means of skeleton in the reactor tank body at this, and those skilled in the art can design as required, as long as can skeleton is fixing in the inner.
Please continue with reference to figure 5 and Fig. 6, it is the synoptic diagram with two steam hose web members that arrange on the upper skeleton.Please in conjunction with Fig. 4, these two steam hose web members 33 and 33 ' are fixedly welded on respectively on the skeleton 31, in this example, steam hose web member 33 is as the exhaust web member, have three ventilation interfaces 331,332 and 333 on it, steam hose web member 33 ' also has three ventilation interfaces 331 ', 332 ' and 333 ' as the air inlet web member on it.And the ventilation interface 331 on the steam hose web member 33 links to each other with silicone tube 35, and silicone tube 35 further links to each other with air inlet transition piece 342 on the sterilization web member 34 in place of using as exhaust; Ventilation interface 331 ' on the steam hose web member 33 ' further links to each other with air inlet transition piece 342 ' on the sterilization web member 34 ' in place that air inlet is used by another silicone tube 35 '.And these ventilation interfaces are outstanding from skeleton 31, so that the winding of steam hose 32.For example, first steam hose 32 is communicated between the ventilation interface 332 and 332 ', and second steam hose 32 is communicated between the ventilation interface 333 and 333 '.Steam hose web member 33 and 33 ' difference only are the length of its main body, this is for the ease of these two steam hose web members 33 and 33 ' being installed in respectively the up and down two ends of skeleton 31, this is because gas is in entering tank body the time, enter from the tank body bottom, the top is sent, and can guarantee the homogeneity of gas flow.For example, steam hose web member 33 ' among Fig. 4 is installed on the bottom of skeleton 31,33 tops that are installed on skeleton 31 of steam hose web member, be inverted in reactor tank body 10 when interior when skeleton 31 like this: steam hose web member 33 ' is positioned at the bottom of tank body 10, is connected with the sterilization web member 34 ' in place that air inlet is used; 33 tops that are positioned at tank body 10 of steam hose web member are connected with the sterilization web member 34 in place that exhaust is used.Yet the present invention is not as limit, and those skilled in the art can arrange the position of steam hose web member as required.In addition, in this implementation column, the ventilation interface quantity that the steam hose web member has is 3, the quantity of steam hose 32 is 2, this is relevant with the reactor size, and reactor is larger usually, needs the steam hose more than 32 that arranges, so ventilation interface quantity is also just more, in actual use, the quantity of the general steam hose 32 of existing reactor is 1 to 4, and the ventilation interface quantity that the steam hose web member has is 2 to 5, yet the present invention is not as limit.
In addition, need to prove, the ventilation interface of steam hose web member, the interface 331,332 and 333 of for example ventilating, identical with the end 3421 of the air inlet transition piece 342 of sterilization web member 34 in place, the one end has the shape of setting one's teeth on edge or helicitic texture (is helicitic texture as shown in the figure,, yet the present invention is not as limit), be conducive to like this its with steam hose 32 and silicone tube 33 between the compactness and the stability that are connected.
Please continue with reference to figure 1, in a preferred embodiment, the tank body of zooblast reactor 10 is interior can to arrange revolving filter 40.Revolving filter 40 is arranged on the transmission shaft 50 usually, and coaxial with transmission shaft 50, to rotate under the driving of transmission shaft 50.The wall of revolving filter 40 arranges microporous membrane, and the pore size of microporous membrane is not specifically limited, as long as so that can not pass through micropore by cultured cells, and the protein of cells produce or virus can be gathered in the crops online by network of micropores and get final product.
Please continue with reference to figure 7, in the present invention's one better implementation column, can be provided with online cell micro observation instrument at the zooblast reactor, with the variation of Real Time Observation cellular form when the animal cell culture.As shown in the figure, be reserved with the reservation socket L1 of online cell micro observation instrument on this zooblast reactor, online cell micro observation instrument is by being connected between flange and the tank body and sealing by the O RunddichtringO, see in detail the patent No. and be 200710042189.0 the online cell micro observation instrument that is used for biochemical reactor, be not described in detail in this.
In the present invention's one better implementation column, can stirring system be set at the zooblast reactor, stirring system can be the mechanical transmission whipping appts, it comprises transmission shaft 50, agitating vane 60 and motor (not shown), agitating vane 60 is installed on the transmission shaft 50, rotated by driven by motor transmission shaft 50 and agitating vane 60, its control mixing speed is usually in the scope of 20～1000r/min, and better control mixing speed is in the scope of 20～250r/min, and what wherein the agitating vane in the mechanical transmission whipping appts 60 was better is wide leaf turbine propulsion formula agitating vane.
In the present invention's one better implementation column, can pH sensor, dissolved oxygen sensor, temperature sensor, pressure transmitter, froth breaking sensor or its combination be set at the zooblast reactor, with the pH value in the sense reactor, dissolved oxygen situation and temperature conditions etc., and then the assurance animal cell culture is carried out under top condition.As shown in the figure, be reserved with the reservation socket L2 of these sensors on this zooblast reactor.
In the present invention's one better implementation column, the microvesicle blow and vent system can also be set in the bottom of zooblast reactor, for example in reservation socket L1 position the microvesicle blow and vent system is set, the ventilation aperture of this microvesicle blow and vent system is 0.5～3 μ m, and the mouth of pipe of its air inlet often links to each other with one or four gas mixing tanks.Four gas mixing tanks are mixed with nitrogen, oxygen, carbonic acid gas and four kinds of gases of air, satisfy the control of animal cell culture process dissolved oxygen and pH.
In the present invention's one better implementation column, also be provided with the tank body LOAD CELLS on the zooblast reactor, as shown in Figure 7, the tank body LOAD CELLS often is arranged at the L3 position on the zooblast reactor.
In the present invention's one better implementation column, also be provided with the feed supplement weighing system on the zooblast reactor, and feed supplement weighing system 70 is feed supplement bottle formula intelligence weighing feeding-system or the cup type intelligent weighing feeding-system of feed supplement.Wherein the cup type intelligent weighing feeding-system of feed supplement is the patent of 200810033769.8 " a kind of supplementary food weighing and metering control device " referring to the patent No., is mainly used in pilot scale and large scale of production.
Please continue referring to Fig. 7, the principle of work of this feed supplement bottle formula intelligence weighing feeding-system is with a kind of supplementary food weighing and metering control device of the patent No. 200810033769.8, comprise feed supplement bottle 71, peristaltic pump 72, respiratory organ 73, LOAD CELLS 74, performer 75 etc., namely with the autocontrol valve in the cup type intelligent weighing feeding-system of peristaltic pump replacement feed supplement, the feed supplement bottle replaces the feed supplement cup, is applicable to the feed supplement of laboratory scale bio-reactor.
The below illustrates the effect of above zooblast reactor take the recombinant animal cell cultivation process as example.Utilize the mentioned online cell micro observation instrument of above implementation column in the mammalian cell reactor, the culturing process of recombinant animal cell (HEK293) to be observed application.Off-line cell blood counting is once carried out in the every 24h sampling of fermenting process.The two cell counting result such as table 2.As shown in Table 2, the cell quantity count results of the blood counting of off-line and online cell micro observation instrument is basically identical.
Table 2 mechanical stir-reactor is cultivated the HEK293 cell density relatively
In sum, more than be used for the zooblast reactor without the bubbles for aeration system, can realize the sterilization in place of the zooblast reactor with bubble-free aerating system of pilot scale and large scale of production.In addition, because bio-reactor of the present invention can be installed revolving filter, stirring system, microvesicle system, bubble-free aerating system simultaneously, it will be appreciated by those skilled in the art that each parts of the present invention and tank body type of attachment are demountable structure, thereby thereby can determine according to the demand of the animal cell culture technique of different varieties whether all or part of employing reaches the best effect of cultivating of zooblast.In addition, the use of multiple sensors of the present invention be so that can carry out correlation analysis to each parameter that reflects the cell physiological characteristic in the culturing process, thereby instructs the optimization of animal cell culture process.Simultaneously, because each parts and the tank body type of attachment of zooblast reactor of the present invention is demountable structure, it will be appreciated by those skilled in the art that on the basis of the present invention this present invention to be combined into the reactor that is applicable to microorganism culturing.
More than show and described ultimate principle of the present invention, principal character and advantage of the present invention.Those skilled in the art should understand; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and the specification sheets is principle of the present invention; the present invention also has various changes and modifications without departing from the spirit and scope of the present invention, and these changes and improvements all fall in the claimed scope of the present invention.The protection domain that the present invention requires is defined by appending claims and equivalent thereof.
Claims (14)
1. the zooblast reactor with bubble-free aerating system comprises reactor tank body and cover, it is characterized in that, also comprise bubble-free aerating system, this bubble-free aerating system comprises:
Skeleton is located in the described reactor tank body;
At least two steam hose web members are located on the described skeleton, and this steam hose web member has at least two ventilation interfaces;
Steam hose is wound on the described skeleton, and is communicated between the ventilation interface of described two steam hose web members;
At least two sterilization web members in place are inserted on the described reactor tank body, and are communicated with a ventilation interface of described steam hose web member respectively;
Described sterilization web member in place comprises:
The balance cavity;
The air inlet transition piece is positioned at an end of described balance cavity, and is communicated with described balance cavity, inserting described reactor tank body, and is communicated with a ventilation interface of described steam hose web member by silicone tube;
Inlet pipe is positioned at the other end of described balance cavity, and is communicated with described balance cavity;
Guide pipe is positioned at described balance cavity, and an end is positioned at described air inlet transition piece and described balance cavity connectivity part, and and described air inlet transition piece between have a space, the other end is arranged in the sidewall of described balance cavity;
Vapor pipe is positioned at the sidewall of described balance cavity, and is communicated with the other end of described guide pipe;
Described balance cavity comprises:
Sheath, described air inlet transition piece is positioned at an end of this sheath, and is communicated with this sheath;
Sleeve pipe, an end is communicated with the other end of described sheath, and the other end is communicated with described inlet pipe, has an opening on its sidewall, and the other end of described guide pipe is arranged in this opening, and described vapor pipe is arranged at this opening part, to be communicated with described guide pipe;
Locking piece is socketed on the described sleeve pipe, and between described sheath and vapor pipe;
Described sheath comprises:
First part, its external diameter linear decrease, and the external diameter smallest end is connected with described air inlet transition piece;
Second section, its external diameter be greater than the maximum outside diameter of first part, and be connected with first part maximum outside diameter end;
Third part, its external diameter are greater than the second section external diameter, and described locking piece is between the third part and vapor pipe of described sheath;
The end of the second section of described sheath and close first part has a groove;
Three parts of described sheath are formed in one.
2. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that the end of described air inlet transition piece has the shape of setting one's teeth on edge.
3. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that, described air inlet transition piece, sheath, sleeve pipe and inlet pipe are one-body molded.
4. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that, the quantity of described steam hose is 1 to 4.
5. described zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that the ventilation interface quantity that described steam hose web member has is 2 to 5.
6. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that described skeleton is provided with fixed support, to be fixedly connected with sealing of tank cover.
7. the zooblast reactor with bubble-free aerating system according to claim 6 is characterized in that, described fixed support is 3 or 4.
8. the zooblast reactor with bubble-free aerating system according to claim 1, it is characterized in that, described skeleton is provided with by taking part, and described reactor tank body inwall is provided with by taking part and supports, described skeleton is fixed in the described reactor tank body by taking part by described.
9. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that, described zooblast reactor is provided with the microvesicle blow and vent system.
10. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that, described zooblast reactor is provided with online cell micro observation instrument.
11. the zooblast reactor with bubble-free aerating system according to claim 1, it is characterized in that described zooblast reactor is provided with the one or more combination in pH sensor, dissolved oxygen sensor, temperature sensor, pressure transmitter or the froth breaking sensor.
12. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that described zooblast reactor is provided with tank body LOAD CELLS or feed supplement weighing system.
13. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that described zooblast reactor is provided with stirring system.
14. the zooblast reactor with bubble-free aerating system according to claim 1 is characterized in that, is provided with revolving filter in the tank body of described zooblast reactor.
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| CN104805013A (en) * | 2015-05-25 | 2015-07-29 | 固元本草汉方生物科技股份有限公司 | Reactor for disposable airlift type plant root tissue culture |
| EP3487979A1 (en) * | 2016-07-19 | 2019-05-29 | The Automation Partnership (Cambridge) Limited | Liquid filtration system with integrated bleed function |
| CN108641958B (en) * | 2018-08-08 | 2024-02-23 | 南京师范大学 | Biological three-dimensional printing environment real-time monitoring and activity maintaining device |
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| DK353585A (en) * | 1984-08-03 | 1986-02-04 | Biotechnolog Forschung Gmbh | PROCEDURE AND APPARATUS FOR BUBBLE GASING OF LIQUIDS, IN PARTICULAR CULTURAL MEDIA FOR PROMOTING TISSUE CULTURES |
| CN1511792A (en) * | 2002-12-31 | 2004-07-14 | 中国科学院生态环境研究中心 | Bubble-free oxygen supply film biological reactor |
| CN1751780A (en) * | 2005-09-02 | 2006-03-29 | 南京工业大学 | Outside circulation gas lift type membrane reactor |
| CN201272799Y (en) * | 2008-09-12 | 2009-07-15 | 广州齐志生物工程设备有限公司 | Biological reactor |
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| DK353585A (en) * | 1984-08-03 | 1986-02-04 | Biotechnolog Forschung Gmbh | PROCEDURE AND APPARATUS FOR BUBBLE GASING OF LIQUIDS, IN PARTICULAR CULTURAL MEDIA FOR PROMOTING TISSUE CULTURES |
| CN1511792A (en) * | 2002-12-31 | 2004-07-14 | 中国科学院生态环境研究中心 | Bubble-free oxygen supply film biological reactor |
| CN1751780A (en) * | 2005-09-02 | 2006-03-29 | 南京工业大学 | Outside circulation gas lift type membrane reactor |
| CN201272799Y (en) * | 2008-09-12 | 2009-07-15 | 广州齐志生物工程设备有限公司 | Biological reactor |
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