CN209722178U - A kind of air-distributing device and the non-static bioreactor comprising it - Google Patents

A kind of air-distributing device and the non-static bioreactor comprising it Download PDF

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Publication number
CN209722178U
CN209722178U CN201920394262.9U CN201920394262U CN209722178U CN 209722178 U CN209722178 U CN 209722178U CN 201920394262 U CN201920394262 U CN 201920394262U CN 209722178 U CN209722178 U CN 209722178U
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air
distributing device
aperture
tank body
static bioreactor
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CN201920394262.9U
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朱义刚
黄涛
董伦海
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Shenzhen Yinglianfeng Biotechnology Co ltd
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Xiamen Heron Biological Technology Co Ltd
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Abstract

The utility model relates to a kind of air-distributing device and include its non-static bioreactor, the air-distributing device includes air accommodating cabinet, its top closure is equipped with multiple equally distributed holes, and described hole at least has 2 kinds of different pore sizes R1, R2, the cap central installs rotatable baffle, and the side wall of the air accommodating cabinet connects inlet channel.The non-static bioreactor includes tank body, and the air-distributing device is mounted in the tank body.Shear stress when ventilation can be effectively reduced in air-distributing device described in the utility model, so that the cell in non-static bioreactor keeps preferable suspended state, high production efficiency, production cost is low, is easy to be mass produced.

Description

A kind of air-distributing device and the non-static bioreactor comprising it
Technical field
The utility model relates to cell cultivation equipment field, especially a kind of air-distributing device and include the non-static of its Bioreactor.
Background technique
Bioreactor is the key equipment that scale carries out cell suspension cultures, by taking culture plant cell as an example, due to The acquiring way of the natural prodcuts of plant origin be primarily now by extracting wild plant resource or artificial growth resource, however Problem of most of wild plant resource due to excessively excavating and facing resource exhaustion.Artificial growth resource is delayed to a certain extent The natural products for having solved plant origin underproduces problem, but to be commonly present content unstable, again for the product in artificial growth source The problems such as metal is exceeded, agriculture is residual exceeded, while artificial growth easily causes the occupancy in forest land, arable land, the destruction problem of environment.Therefore Using formation for Plant Cell Suspension technology, the industrialized production of natural active product is carried out in conjunction with bioreactor technology, is to solve to plant One of the important channel of the natural products shortage of resources in object source.
Plant cell suspension culture bioreactor is sent out on the basis of traditional microbiological culture bioreactors at present Zhan Erlai, main Types have mechanical agitation type, gas-lifting type, bubble type, rotary drum and fixed bed etc..The structure of the above bioreactor Type is cylindrical structure, finds that cylinder configuration bioreactor bottom is easy deposition when plant cell suspension culture in research Compared with many cells, it is easy to appear building-up effect, causes depositional phenomenon serious especially plant cell suspension culture early period and later period cell, Growth and breeding and the secondary metabolite synthesis for directly affecting cell, cause cell-unit matrix yield and target product content low The problem of.Although agitating paddle can improve deposition to a certain extent, susceptibility ratio of the cell mass assembled to shearing force It is unicellular high more, so that a large amount of cells are dead during the culture that suspends.Gas-lifting type and Sparged bioreactors are in order to make to gather The cell mass of collection suspends, and needs sufficiently large ventilation ratio, however shearing force caused by bubbles burst while ventilation ratio increase It greatly increases, while increasing with foam, carbon dioxide is excessively escaped, and the growth and metabolism of plant cell are unfavorable for.Immobilization Culture reduces plant cell to the easy differentiating phenomenon of sensitivity and cell of shearing force to a certain extent, but natural products belongs to more In intracellular product, its popularization and application is limited.
Disposable wave bioreactor is the relatively suitable animal and plant cells suspension culture to grow up the nearly more than ten years Bioreactor, mainly by shake platform make wave effect increase gas-liquid area reach mixing and oxygen supply.But It is found in research, disposable wave bioreactor is directed to suspension culture effect easily assemble, the biggish cell mass of particle It is poor, and coefficient generally only has 50%.Chinese invention patent CN103224882A is disclosed with the primary of aerator Property wave bioreactor, but for the plant cell aggregates easily assembled, increasing aerator, the effect is unsatisfactory, Show as only having certain impetus in narrow range around the aerator, if increasing ventilatory capacity, shearing force and bubble Foam problem also increases, and is unfavorable for plant cell growth breeding, therefore bottom plane area is excessive for plant cell suspension culture It is extremely unfavorable.
Utility model content
For above situation, the utility model provides a kind of air-distributing device, when solving cell suspension cultures ventilation Bring shearing force is excessive, the problem that bring liquid mixed effect is poor, culture efficiency is low.
The present invention also provides a kind of non-static bioreactor comprising the air-distributing device, which, which suspends, is cultivated Effect is good, and oxygen mass transfer coefficients are suitable for that shearing force is suitable for that cost is relatively low, is easy to carry out mass cell suspension culture.
Air-distributing device described in the utility model, by the way of the hole combination of different pore size, in different cultures Phase is rotated with baffle and carries out aperture switching, specifically: when progenitor cells density is lower, using larger aperture, later period density compared with Smaller aperture due is used when high, to realize in the certain ventilation ratio of holding or in the case where being finely adjusted, meets different growths Demand of the phase cell to dissolved oxygen, while effectively avoiding in the prior art in order to change dissolved oxygen amount, it is simple by increase ventilation Than come the phenomenon that realizing, cause gas excessive to the shearing force of cell, lead to cell death.In addition, the air-distributing device Adjusting to dissolved oxygen amount is realized using the thinking in switching aperture, is effectively avoided because of big ventilation bring foam proliferation issues, it can So that coefficient is promoted by the 50% of the prior art to 80% or more.Finally, the biology using the air-distributing device is anti- Answer device, can effectively save air cost, unit base cell yield can achieve the high density of 15g/l or more.
To solve this technical problem, the utility model takes following scheme:
A kind of air-distributing device, including air accommodate cabinet, and the air accommodates cabinet sealed bottom, on top closure Equipped with multiple equally distributed holes, and described hole at least has 2 kinds of different pore sizes R1, R2, and the cap central installation can The side wall of the baffle of rotation, the air accommodating cabinet connects air hose.
Further, the air-distributing device is one of plastics, silica gel, stainless steel or titanium alloy material or several The combination of kind.
Further, the 10um≤aperture aperture R1 < R2≤1000um.
Further, the 100um≤aperture aperture R1 < R2≤500um, and aperture R1: aperture R2=1:1.5-3.5.
Further, the fan-shaped distribution of the hole of different pore size, and the sector of same apertures composition is in the capping Heart-shaped in a center of symmetry, the sector of different pore size composition is adjacent to each other.
Further, the baffle includes multiple sectors, and the sector is centrally formed pair with the capping Claim distribution, the edge of the sector is no more than the outer of the capping.
Based on the same inventive concept, the utility model also provides a kind of non-static bioreactor, including tank body, the tank Inoculation mouth, culture medium entrance and gas outlet are equipped with above body, the tank base is equipped with air inlet, the fixed peace of the tank interior The air-distributing device is filled, the air inlet is connected with the air hose on the air-distributing device side wall.
Further, the tank body is flexible material, and the tank base has inclined surface, the air-distributing device peace On the inclined surface.
Further, packing density of the air-distributing device on the inclined surface is 20-60/square meter.
Further, packing density of the air-distributing device on the inclined surface is 35-45/square meter.
Air-distributing device described in the utility model compared with prior art, by switching air-distributing device in not Same aperture effectively reduces ventilation bring shear stress to adapt to the dissolved oxygen amount demand of different cultivation stages.
Non-static bioreactor construction comprising the air-distributing device is succinct, high production efficiency, and unit base is thin Born of the same parents' yield can achieve the high density of 15g/l or more;It effectively avoids to make because of big ventilation bring foam proliferation issues simultaneously Coefficient is promoted by the 50% of the prior art to 80% or more.
Detailed description of the invention
Fig. 1 is the air-distributing device structural schematic diagram that the utility model embodiment 1 provides;
Fig. 2 is the non-static bioreactor construction schematic diagram that the utility model embodiment 2 provides.
Specific embodiment
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, the embodiments are a part of the embodiments of the present invention, without It is whole embodiments, is merely to illustrate the utility model, and is not construed as limitation the scope of the utility model.It is practical based on this Embodiment in novel, every other reality obtained by those of ordinary skill in the art without making creative efforts Example is applied, is fallen within the protection scope of the utility model.The person that is not specified actual conditions in embodiment, according to normal conditions 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 For ease of description the utility model and simplify description, rather than the device or element of indication or suggestion meaning must have it is specific Orientation, be constructed and operated in a specific orientation, therefore should not be understood as limiting the present invention.In addition, term " the One ", " second ", " third " are used for descriptive purposes only and cannot be understood as indicating or suggesting 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, be also possible to be electrically connected;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, above-mentioned art can be understood with concrete condition The concrete meaning of language in the present invention.
Embodiment 1:
With reference to Fig. 1, a kind of air-distributing device, including air accommodating cabinet 71 and air hose 72, air hose 72 connect sky Gas accommodates the side wall of cabinet 71, and filtrated air is sent into inside air accommodating cabinet 71.Air accommodates 71 sealed bottom of cabinet, top Portion's capping is equipped with multiple equally distributed holes, and aperture is respectively R1, R2, and wherein the aperture of aperture 73 is R1, the hole of macropore 74 Diameter is R2, aperture 73 and the respectively fan-shaped aggregation of macropore 74, with the central symmetry that is centrally formed of the air accommodating capping of cabinet 71, and Aperture 73 is formed by fan-shaped and macropore 74 and is formed by sector adjacent to each other.
Preferably, the 10um≤aperture aperture R1 < R2≤1000um is realized under suitable ventilation ratio to dissolved oxygen with meeting The adjustment of amount.It is highly preferred that 100um≤aperture the aperture R1 < R2≤500um, and aperture R1: aperture R2=1:1.5-3.5, this When suspension culture effect it is good, oxygen mass transfer coefficients be suitable for, shearing force be suitable for.
Air accommodates 71 cap central of cabinet and installs rotatable baffle 75, and baffle 75 includes multiple sectors, fan-shaped Blade is symmetrical with being centrally formed for covering, and the edge of sector and the outer of capping are concordant.It is arrived by butterfly 75 Specific position can carry out the switching between the ventilation of macropore 74 or the ventilation of aperture 75.The mode of switching control belongs to well-known technique, This will not be repeated here.
Embodiment 2:
A kind of non-static bioreactor, with reference to Fig. 2, including the tank body being made of the first cavity 1, the second cavity 2, In, the first cavity 1 is located at 2 top of the second cavity.First cavity 1 is cylinder, and the second cavity 2 is back taper, and the second cavity A side horizontal by 90 ° of angles, another side is horizontal by 45 ° of angles (i.e. 45 ° of inclined surface inclination angle).First The base circumference of the base circumference of cavity 1 and the second cavity 2 is overlapped, and two cavitys connect.The material of tank body is preferably polyamides Amine or polyethylene or ethylene-vinyl acetate copolymer or ethylene-vinyl alcohol copolymer.
First cavity 1 is equipped with inoculation mouth 3, culture medium entrance 5 and gas outlet 4, connects piping connection.Wherein, preferably It is connected on 1 top closure of the first cavity, convenient for inoculation and charging.Control and adjustment to tank interior air pressure for convenience, Respirator 41 and the first air control valve 42 are set on the pipeline of connection gas outlet 4, so that the intracorporal gas of tank is through the first sky Gas regulating valve 42 enters respirator 41 after adjusting flow.
Multiple air-distributing devices 7, the air hose 72 of all air-distributing devices 7 are installed on the inclined surface of second cavity 2 It is connected, and is connected with air inlet 6, control charge flow rate for convenience, flowmeter 61 and second is equipped on air inlet pipeline Air control valve 62 so that filtrated air is after flowmeter 61, using the second air control valve 62 regulation and control into Enter the second cavity 2.
Further, 1 side wall of the first cavity is equipped with sensor reserved opening 8, for being inserted into pH meter, air saturation detection dress It sets or device for detection of carbon dioxide content, such as dissolved oxygen meter.
The present embodiment is only exemplary, and the utility model is not limited to such structure, and those skilled in the art should think It arrives, the second cavity 2 can also have other inclined surface structures, such as inverted pyramid, or the inverted cone with symmetry axis.Second cavity 2 inclined surface inclination angle can also be other numerical value, such as 30 °, 40 °, 50 °, 60 ° or 70 °, inclination angle difference will affect suspension State can be adjusted according to the density difference of institute's Object of Development.Using the tank body with inclined surface, it is possible to prevente effectively from existing It precipitates to form plane because aggregation is agglomerating in cell cultivation process in technology, so that entire cultivation cycle inner cell be made to be in good Suspended state is conducive to cell growth and breeding, improves unit base cell yield, reduces production cost.
Embodiment 3
The present embodiment is suspended by ginseng-cell for culture, describes the user of non-static bioreactor in embodiment 2 Method:
By ginseng-cell callus aseptic inoculation in the qualified fluid nutrient medium of sterilizing, it is placed in 22-25 DEG C, 80- Qualified loblolly shape ginseng liquid seeds are passed through the sterile access of inoculation mouth 3 by shake culture 15-25 days in 120rpm rotary shaker Pass through in the non-static bioreactor tank body of suffocating sterilization or irradiation sterilization in advance, while through peristaltic pump that sterilizing is qualified Ginseng-cell fluid nutrient medium is accessed in tank body by culture medium entrance 5, and filtrated air is inputted in tank body by air inlet 6, Tank body is discharged by gas outlet 4 in filtrated air simultaneously, adjusts tank by the second air control valve 62 and presses micro-positive pressure, is placed in constant temperature Room controls 22-25 DEG C of cultivation temperature, controls ventilation ratio in 0.12-0.24vvm by flowmeter 61.
Air-distributing device 7 is assembled into tank body before tank body sterilizing, and in 74 gears, (i.e. baffle 75 covers in aperture for adjustment 73, expose macropore 74).Packing density of the air-distributing device 7 on inclined surface is 20-60/square meter, preferably 35-45/ Square meter.Culture is ventilated early period using macropore 74, and culture density increase is gradually adjusted to aperture 73 and ventilated.According to naked eyes during culture Observe cell concentration variation, or the relative atmospheric saturation degree and titanium dioxide of the electrode detection by being inserted into sensor reserved opening 8 Carbon amounts gradually adjusts macropore 74 to 73 gear of aperture.Entire culture period suspension 20-40 days, during which sample detection cell concentration, reached Fermentation can be terminated to 15g/l or more, filter cake low-temperature vacuum drying is harvested to obtain finished product by all fermentation liquid centrifugal filtrations, is calculated Unit base cell yield is 15.78g/l.
Although the embodiments of the present invention have been shown and described above, it is to be understood that above-described embodiment is Illustratively, it should not be understood as limiting the present invention, those skilled in the art are not departing from the utility model Principle and objective in the case where above-described embodiment can be changed in the scope of the utility model, modify, replace and Modification.

Claims (10)

1. a kind of air-distributing device, including air accommodate cabinet, the air accommodates cabinet sealed bottom, sets on top closure There are multiple equally distributed holes, and described hole at least has 2 kinds of different pore sizes R1, R2, the cap central installation can revolve The side wall of the baffle turned, the air accommodating cabinet connects air hose.
2. air-distributing device according to claim 1, it is characterised in that: the air-distributing device be plastics, silica gel, One of stainless steel or titanium alloy material or several combinations.
3. air-distributing device according to claim 1, it is characterised in that: the 10um≤aperture aperture R1 < R2≤ 1000um。
4. air-distributing device according to claim 3, it is characterised in that: the 100um≤aperture aperture R1 < R2≤ 500um, and aperture R1: aperture R2=1:1.5-3.5.
5. air-distributing device according to claim 1, it is characterised in that: the fan-shaped distribution of the hole of different pore size, and The fan-shaped central symmetry that is centrally formed with the capping of same apertures composition, the sector of different pore size composition are adjacent to each other.
6. air-distributing device according to claim 1 or 5, it is characterised in that: the baffle includes multiple sectors, The sector is symmetrical with being centrally formed for the capping, and the edge of the sector is no more than the outer of the capping Edge.
7. a kind of non-static bioreactor, including tank body, the tank body top is equipped with inoculation mouth, culture medium entrance and outlet Mouthful, the tank base is equipped with air inlet, and the distribution of air described in any one of claim 1-6 is fixedly mounted in the tank interior Device, the air inlet are connected with the air hose on the air-distributing device side wall.
8. non-static bioreactor according to claim 7, it is characterised in that: the tank body is flexible material, described Tank base has inclined surface, and the air-distributing device is mounted on the inclined surface.
9. non-static bioreactor according to claim 8, it is characterised in that: the air-distributing device inclines described Packing density on inclined-plane is 20-60/square meter.
10. non-static bioreactor according to claim 9, it is characterised in that: the air-distributing device is described Packing density on inclined surface is 35-45/square meter.
CN201920394262.9U 2019-03-27 2019-03-27 A kind of air-distributing device and the non-static bioreactor comprising it Active CN209722178U (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110903949A (en) * 2019-12-30 2020-03-24 上海复旦张江生物医药股份有限公司 Ventilation mechanism and biological fermentation device

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110903949A (en) * 2019-12-30 2020-03-24 上海复旦张江生物医药股份有限公司 Ventilation mechanism and biological fermentation device

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Effective date of registration: 20231007

Address after: 518000, 401, No. 36 Dadongmen Street, Xinlan Community, Guanlan Street, Longhua District, Shenzhen City, Guangdong Province

Patentee after: Shenzhen Yinglianfeng Biotechnology Co.,Ltd.

Address before: 1st and 4th floors of Building 11, Biopharmaceutical General Factory, No. 2020 Wengjiao West Road, Haicang District, Xiamen City, Fujian Province, 361000

Patentee before: XIAMEN AMOY HERON BIOTECHNOLOGY CO.,LTD.

TR01 Transfer of patent right