CN207958395U - A kind of bioreactor benefit alkali equipment - Google Patents
A kind of bioreactor benefit alkali equipment Download PDFInfo
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- CN207958395U CN207958395U CN201820130749.1U CN201820130749U CN207958395U CN 207958395 U CN207958395 U CN 207958395U CN 201820130749 U CN201820130749 U CN 201820130749U CN 207958395 U CN207958395 U CN 207958395U
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- agitating
- agitating paddle
- lye
- tank
- bioreactor
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Abstract
The utility model is related to animal cell culture technology fields, and alkali equipment is mended in particular to a kind of bioreactor；The equipment includes tank body；The tank interior is provided with agitating device；Lye injector head is provided with above the tank skin of the tank body, the lye injector head is connected by valve with storage alkali container.The utility model by using pulsed lye additional way, the agitating device and baffle that collocation is especially arranged, the generation of turbulent flow, quick mixing lye when can effectively reduce stirring, to reduce the cell death caused by mechanical damage and local concentration of lye are excessively high.In addition, surprisingly, unique agitating paddle rotation mode can also extend the residence time of oxygen, to effectively reduce the consumption of oxygen.
Description
Technical field
The utility model is related to animal cell culture technology fields, and mending alkali in particular to a kind of bioreactor sets
It is standby.
Background technology
Cell culture starts from the beginning of this century, starts to expand to scale in 1962, is developed so far and has become in biology, medicine
Widely used technology in research and application utilizes enzyme, growth factor, vaccine of the cell culture production with important medical value
With monoclonal antibody etc., have become the pith of medical biotechnology industry.
The accumulation of metabolic waste is cytostatic principal element in cellular environment, these metabolic wastes include dioxy
Change carbon, lactic acid and ammonia.Wherein, carbon dioxide and lactic acid are most important acidic metabolic wastes in culture medium, with cell culture
The increase of scale and density, carbon dioxide and lactic acid are built up in cell culture medium, are produced to the normal physiological function of cell
It is raw to inhibit even toxic action.
In the prior art mainly by adding alkaline matter (such as NaHCO into cell culture fluid3Solution) it neutralizes
Lactic acid is added in tank (as shown in Figure 1) using the method that no pressure is added dropwise along tank skin stream.This fed-batch mode is not only slow, but also
Cannot rapidly dissolve and come after being added drop-wise in tank, and cause lye surrounding cell total loss and dead, the waste of lye
Also very big.
In view of this, special propose the utility model.
Utility model content
The utility model is related to a kind of bioreactors to mend alkali equipment, including tank body；
The tank interior is provided with agitating device；
Lye injector head is provided with above the tank skin of the tank body, the lye injector head passes through valve and storage alkali container phase
Even.
Device provided by the utility model can ensure that the stirring of lye prestissimo is opened, to reduce the dead of cell
Die quantity.
The utility model further relates to a kind of bioreactor alkali supplement method, is carrying out mending alkali behaviour using equipment as described above
It is intermittent to open the valve when making so that the lye injector head pulsed sprays lye, opens simultaneously the agitating device
So that mixing is complete rapidly for the lye of supplement.
Description of the drawings
It, below will be right in order to illustrate more clearly of specific embodiment of the present invention or technical solution in the prior art
Specific implementation mode or attached drawing needed to be used in the description of the prior art are briefly described, it should be apparent that, it is described below
In attached drawing be that some embodiments of the utility model are not paying creativeness for those of ordinary skill in the art
Under the premise of labour, other drawings may also be obtained based on these drawings.
Fig. 1 is the benefit alkali equipment principle schematic diagram of the prior art；
Fig. 2 is that alkali equipment principle schematic diagram is mended used by the utility model；
Fig. 3 is the structural schematic diagram of the benefit alkali equipment employed in the utility model one embodiment；
Fig. 4 is the schematic top plan view of the benefit alkali equipment employed in the utility model one embodiment.
Reference numeral：
Tank body A；
Microvesicle air-breather 1
Agitating device 2；Agitating shaft 201；Agitating paddle 202；
Power plant 3；
Baffle 4；
Lye injector head 5；Valve 501.
Specific implementation mode
The technical solution of the utility model is clearly and completely retouched below in conjunction with the drawings and specific embodiments
It states, it will be appreciated by those skilled in the art that following described embodiment is the utility model a part of the embodiment, without
It is whole embodiment, is merely to illustrate the utility model, and is not construed as limitation the scope of the utility model.Based on this practicality
Embodiment in novel, every other reality obtained by those of ordinary skill in the art without making creative efforts
Example is applied, shall fall within the protection scope of the present invention.The person that is not specified actual conditions in embodiment, according to normal condition or manufacturer
It is recommended that condition carry out.Reagents or instruments used without specified manufacturer is the routine that can be obtained by commercially available purchase
Product.
It is in the description of the present invention, it should be noted that term "center", "upper", "lower", "left", "right", " perpendicular
Directly ", the orientation or positional relationship of the instructions such as "horizontal", "inner", "outside" is to be based on the orientation or positional relationship shown in the drawings, and is only
The utility model and simplifying describes for ease of description, do not indicate or imply the indicated device or element must have it is specific
Orientation, with specific azimuth configuration and operation, therefore should not be understood as limiting the present invention.In addition, term " the
One ", " second ", " third " are used for description purposes only, and are not understood to indicate or imply relative importance.
In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " is pacified
Dress ", " connected ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or integrally
Connection；It can be mechanical connection, can also be electrical connection；Can be directly connected, can also indirectly connected through an intermediary,
It can be the connection inside two elements.For the ordinary skill in the art, it can understand above-mentioned art with concrete condition
The concrete meaning of language in the present invention.
The utility model is related to a kind of bioreactors to mend alkali equipment, as shown in figure 3, the equipment includes tank body A；
The tank body A is internally provided with agitating device 2；
Lye injector head 5 is provided with above the tank skin of the tank body A, the lye injector head 5 passes through valve 501 and storage alkali
Container is connected.
The principle of the equipment is as shown in Fig. 2, lye is spurted into the culture medium of tank body with pulsed by lye injector head 5
It is interior, quickly stirred and evenly mixed in agitating device 2, can to avoid local pH it is excessively high caused by cellular damage.
Preferably, the bioreactor is mended in alkali equipment and is additionally provided with pH meter.
Preferably, bioreactor as described above mends alkali equipment, and 5 front end endoporus aperture of the lye injector head is 1mm
~3mm；More preferably 2mm.
Preferably, bioreactor as described above mends alkali equipment, and the agitating device 2 includes agitating shaft 201 and stirring
Paddle 202；
The agitating shaft 201 is perpendicular to horizontal plane and is fixed on tank bottom, and the agitating paddle 202 is set to the agitating shaft
On 201, the bottom of the agitating shaft 201 is connected with power plant 3；
Preferably, the equipment further includes microvesicle air-breather 1, and the microvesicle air-breather 1 is set to the agitating paddle
202 underface.
Microvesicle air-breather 1 can be commercial product in the present invention, can be to cell oxygen supply/CO2.
Preferably, bioreactor as described above mends alkali equipment, and there are three the agitating paddles 202, from the agitating shaft
From top to bottom it is followed successively by the first agitating paddle, the second agitating paddle and third agitating paddle；The second agitating paddle agitated liquid is by upper
It moves downward；The first and third agitating paddle agitated liquid moves from bottom to top；
By above-mentioned set-up mode, the oxygen discharged in microvesicle air-breather 1 can be confined between the first and third agitating paddle
(it is considered that spiral liquid stream is formd between three agitating paddles), it is resident in culture solution to extend bubbles of oxygen
Time increases the utilization rate of oxygen.
Preferably, the agitating paddle 202 is turbine type agitating paddle or pusher agitating paddle；
It is furthermore preferred that the agitating paddle 202 is four leaf push type agitating paddles.
Preferably, the rake angle degree of the agitating paddle is 40 °~60 °；More preferably 50 °；Since each agitating paddle makes
The direction of motion of liquid stream is different, thus the deflection of the deflection angle and first and third agitating paddle of preferred second agitating paddle
Angle mirror symmetry.
Preferably, the ratio of the paddle diameter of the agitating paddle and tank diameter is 0.2~0.7, more preferably 0.3~0.5.
Preferably, the rotating speed of the agitating paddle is 30~70rpm.
Preferably, bioreactor as described above mends alkali equipment, and baffle 4 is additionally provided with inside the tank skin of the tank body,
The baffle 4 is perpendicular to horizontal plane；
Preferably, the baffle 4 has 3~5, more preferably 4, and is uniformly distributed on the tank skin of the tank body；
Preferably, as shown in figure 4, the baffle 4 deviate its with 30 ° of Grain lines where the tank body A tie points~
40 °, and each baffle deflection direction is consistent；
Preferably, the width of the baffle is 1/13~1/10 tank diameter.
It is furthermore preferred that each baffle deflection direction is consistent with the direction of rotation of agitating paddle；Such as the direction of rotation of agitating paddle is
Counterclockwise, then the deflection direction of the baffle 4 just as shown in figure 4, being deflected in obtuse angle along flow counterclockwise.
Due to second agitating paddle and first and third agitating paddle by the direction of aqueous agitation on the contrary, culture solution into
When row agitation liquid flow direction can than three agitating paddles equidirectional (upwards agitation) it is more chaotic, therefore, this agitation mode
Although dissolved oxygen amount can be greatly increased, it is also easier to, because the chaotic impact of liquid stream forms turbulent flow, it is broken in turn result in cell simultaneously
Broken death.Based on this reason, the utility model is provided with baffle 4, and its deflection angle is consistent with the direction of rotation of agitating paddle, gear
Plate 4 can play the role of water conservancy diversion, since cell culture fluid closes on flow velocity maximum at tank skin in bioreactor, thus on tank skin
Baffle 4 can slow down flow rate of liquid, guiding liquid flow direction herein, and then effectively avoid the generation of turbulent flow.
The utility model further relates to a kind of bioreactor alkali supplement method, is carrying out mending alkali behaviour using equipment as described above
It is intermittent to open the valve 501 when making so that 5 pulsed of lye injector head sprays lye, opens simultaneously the stirring
Device 2 is so that the lye of supplement mixes rapidly completely.
Preferably, the tank body A is 3000L~6000L volumes；
Preferably, bioreactor alkali supplement method as described above, the lye are 0.8mol/L~1.2mol/L's
NaHCO3Solution；The more preferably NaHCO of 1mol/L3Solution.
Preferably, bioreactor alkali supplement method as described above, the valve 501 open one at interval of 4.5s~5.5s
It is secondary, 50ms~200ms is opened every time；It is furthermore preferred that the valve 501 opened at interval of 5s it is primary, open every time 100ms~
150ms。
Preferably, bioreactor alkali supplement method as described above, when spraying lye, the liquid in the lye injector head 5
Body pressure is 0.08MPa~0.12MPa；It is furthermore preferred that the fluid pressure in the lye injector head 5 is 0.10MPa.
Preferably, bioreactor alkali supplement method as described above, when carrying out mending alkali operation, the agitating device 2
Rotating speed is 30rpm~70rpm；It is furthermore preferred that the rotating speed of the agitating device 2 is 50rpm.
The switch time of concentration of lye, valve, the rotating speed of agitating device and pressure (the cooperation injection in lye injector head
The outlet diameter of head, what is actually limited is the lye amount sprayed every time) it is that applicant combines the biology of the utility model anti-
Answer device to mend alkali device structure especially to set, to can quick mixing lye, to reduce its injury to cell.
Wherein, the opening and closing of the valve, agitating device rotating speed can be controlled by electronic control unit, control dress
It sets by reading pH meter data and analyzing, and then controls the operating of above-mentioned apparatus.
Embodiment
In the present embodiment, bioreactor tank body is 6000L tank bodies, cultivates floating type mdck cell.Above tank skin
It is provided with lye injector head, lye injector head passes through valve and storage 1.0mol/L NaHCO3The container of solution is connected, and valve is every
It is spaced 5s to open once, opens 120ms every time.When spraying lye, the fluid pressure in the lye injector head is 1.0MPa, alkali
The a diameter of 2mm of jet port of liquid injector head.Bottom is provided with microvesicle air-breather for leading to oxygen in tank, and bottom is also set up in tank
It includes agitating shaft and agitating paddle to have agitating device, the agitating device, and the agitating shaft is perpendicular to horizontal plane and is fixed on tank bottom,
The agitating paddle is set on the agitating shaft, and the bottom of the agitating shaft is connected with power plant；Three are arranged on agitating shaft
Agitating paddle is from top to bottom followed successively by the first agitating paddle, the second agitating paddle and third agitating paddle, is to rotate clockwise；Described
Two agitating paddle agitated liquids move from top to bottom；The first and third agitating paddle agitated liquid moves from bottom to top.The paddle of agitating paddle
Leaf angle of inclination is 50 °, and the ratio of paddle diameter and tank diameter is 0.4.When lye is added dropwise, rotating speed of agitator is 50 turns/min；Normally
When culture cell leads to oxygen, rotating speed of agitator is 40 turns/min.Four baffles are set on tank skin, and the deflection direction of baffle is up time
Needle deviates itself and 35 ° of Grain lines where the tank body tie point, and the width of baffle is 1/12 tank diameter.It sets this embodiment to
Comparative example is arranged in experimental group on this basis.
Comparative example 1：It is differed only in experimental group, removes baffle.
Comparative example 2：Lye injector head is removed, add alkaline methods replace with side wall in the prior art and formula is added dropwise；Agitating paddle
Rotating speed is always 40 turns/min.
Comparative example 3：It is differed only in experimental group, baffle is arranged perpendicular to tank body.
Comparative example 4：It is differed only in experimental group, three agitating paddles stir culture solution upwards, and remove baffle.
It is that 1,800,000/ml is cultivated with batch mdck cell 5 days by cell culture density, by adding alkali to maintain, pH is constant to be
6.8-7.2 detects following data：
From comparative example 1 and 2 it is found that mode is added dropwise in different lye has large effect to Cell viability.Adherent dropwise addition drop
The mode added is also continuous, and it is excessive to often result in concentration of lye at dropwise addition, thus can cause more cell death.If accelerating
Rotating speed then also results in the increase that mechanical damage causes cell mortality；Also can not in addition, dosage is added dropwise in adherent dropwise addition every time
Control, can not precisely adjust pH.From comparative example 1 and 3 it is found that baffle plate setting mode it is incorrect or removal baffle, although can reduce
Flow of oxygen, but more violent turbulent flow can be formed, cause Cell viability to be greatly lowered.By comparative example 4 it is found that three stirrings
When paddle stirs culture solution upwards, it is resident and is shortened in culture solution due to oxygen, thus oxygen utilization rate declines, oxygen consumption total amount
It is soaring apparent；It is uniformly dispersed in addition, the change of aqueous agitation mode is also unfavorable for oxygen, since the utility model uses big volume
Bioreactor, the mixing difficulty of oxygen is consequently increased, it is possible to which the decline for causing local oxygen concentration causes cell to be lived
Rate reduces.
In conclusion lye additional way of the utility model by using pulsed, the stirring dress being especially arranged of arranging in pairs or groups
Set and baffle, can effectively less stirring when the generation of turbulent flow, quick mixing lye, to reduce because of mechanical damage and local alkali
Cell death caused by liquid excessive concentration.In addition, surprisingly, unique agitating paddle rotation mode can also extend oxygen
Residence time, to effectively reduce oxygen consumption.
Finally it should be noted that：The above various embodiments is only to illustrate the technical solution of the utility model, rather than limits it
System；Although the utility model has been described in detail with reference to the foregoing embodiments, but those skilled in the art answer
Work as understanding：It still can be with technical scheme described in the above embodiments is modified, either to which part or entirely
Portion's technical characteristic carries out equivalent replacement；And these modifications or replacements, this practicality that it does not separate the essence of the corresponding technical solution
The range of novel each embodiment technical solution.
Claims (10)
1. a kind of bioreactor mends alkali equipment, which is characterized in that including tank body；
The tank interior is provided with agitating device；
Lye injector head is provided with above the tank skin of the tank body, the lye injector head is connected by valve with storage alkali container.
2. bioreactor according to claim 1 mends alkali equipment, which is characterized in that lye injector head front end endoporus
Aperture is 1mm~3mm.
3. bioreactor according to claim 1 mends alkali equipment, which is characterized in that the agitating device includes agitating shaft
And agitating paddle；
The agitating shaft is perpendicular to horizontal plane and is fixed on tank bottom, and the agitating paddle is set on the agitating shaft, the stirring
The bottom of axis is connected with power plant.
4. bioreactor according to claim 3 mends alkali equipment, which is characterized in that the equipment further includes microvesicle ventilation
Device, the microvesicle air-breather are set to the underface of the agitating paddle.
5. bioreactor according to claim 3 mends alkali equipment, which is characterized in that there are three the agitating paddles, from institute
It states agitating shaft and is from top to bottom followed successively by the first agitating paddle, the second agitating paddle and third agitating paddle；The second stirring paddle stirring
Liquid moves from top to bottom；The first and third agitating paddle agitated liquid moves from bottom to top.
6. bioreactor according to claim 3 mends alkali equipment, which is characterized in that the agitating paddle stirs for turbine type
Paddle or pusher agitating paddle.
7. bioreactor according to claim 3 mends alkali equipment, which is characterized in that the agitating paddle is that four leaves are pusher
Agitating paddle.
8. mending alkali equipment according to claim 1~7 any one of them bioreactor, which is characterized in that the tank of the tank body
Pars intramuralis is additionally provided with baffle, and the baffle is perpendicular to horizontal plane.
9. bioreactor according to claim 8 mends alkali equipment, which is characterized in that the baffle has 3~5, and
It is even to be distributed on the tank skin of the tank body.
10. bioreactor according to claim 9 mends alkali equipment, which is characterized in that the baffle deviates itself and institute
30 °~40 ° of Grain lines where tank body tie point are stated, and each baffle deflection direction is consistent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820130749.1U CN207958395U (en) | 2018-01-25 | 2018-01-25 | A kind of bioreactor benefit alkali equipment |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820130749.1U CN207958395U (en) | 2018-01-25 | 2018-01-25 | A kind of bioreactor benefit alkali equipment |
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| CN207958395U true CN207958395U (en) | 2018-10-12 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108048323A (en) * | 2018-01-25 | 2018-05-18 | 吉林冠界生物技术有限公司 | A kind of bioreactor mends alkali device and method |
-
2018
- 2018-01-25 CN CN201820130749.1U patent/CN207958395U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108048323A (en) * | 2018-01-25 | 2018-05-18 | 吉林冠界生物技术有限公司 | A kind of bioreactor mends alkali device and method |
| CN108048323B (en) * | 2018-01-25 | 2024-02-06 | 吉林冠界生物技术有限公司 | Alkali supplementing equipment and method for bioreactor |
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