CN111676136A - Bioreactor for improving antigen yield - Google Patents
Bioreactor for improving antigen yield Download PDFInfo
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- CN111676136A CN111676136A CN202010582982.5A CN202010582982A CN111676136A CN 111676136 A CN111676136 A CN 111676136A CN 202010582982 A CN202010582982 A CN 202010582982A CN 111676136 A CN111676136 A CN 111676136A
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
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- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/44—Means for regulation, monitoring, measurement or control, e.g. flow regulation of volume or liquid level
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Abstract
The invention discloses a bioreactor for improving antigen yield, which comprises: the device comprises a closed tank-shaped shell, a stirring device, a progressive spiral pipe for stirring, a rotating shaft, a storage device, an inverted triangular shell of the storage device, a flow guide screen plate, a fluid circulating pipeline, a fluid circulating pump, a deposition box body at the bottom of the progressive spiral pipe and a liquid level sensor. By using the technology, the culture volume of the antigen cells is larger than that of the existing bioreactor.
Description
Technical Field
The invention relates to a bioreactor, in particular to a bioreactor for improving the antigen yield, belonging to the field of antigen cell culture.
Background
The avian influenza epidemic first appeared in italy at 1878, when a number of flock deaths occurred, known as fowl plague. In 1995, scientists demonstrated that the causative virus responsible for the epidemic situation of avian influenza was influenza a virus, and at that time, the spread of the virus was mainly prevented by a method of sterilizing, isolating, and slaughtering livestock in large quantities. In 1999, human infections with avian influenza type H9N2 occurred in hong kong, leading to death of the people. According to statistics, the number of people infected with H5N1 in the whole world is about 624 people and 371 people die in the year 2003-2013.
The recombinant live vector vaccine is prepared by using the antigenic gene with immunogenicity of the avian influenza virus and weak pathogenicity virus or bacteria as a vector to construct recombinant virus or bacteria capable of expressing the antigenic gene of the avian influenza virus, so that the recombinant virus or bacteria can be replicated and expressed in an animal body, and an animal body is induced to generate a stronger immune response reaction.
The cell culture technology of the antigen is born in the 60 th 20 th century, is a technology for producing biological products in a large scale and high efficiency by using a bioreactor under artificial conditions, has the advantages of high production efficiency, simple operation, easy quality and biological safety control and the like, is widely applied to the field of biological products, has wider popularization and application in the production of antigen vaccines, is an advanced technology in the field of the current antigen vaccine production, and is also a development direction of the future antigen vaccine production.
The existing stirring bioreactor has the technical problems of low stirring efficiency and even poor antigen cell culture efficiency in the technical requirement of high-density large-scale cell culture, which is the bottleneck of large-scale cell culture.
Therefore, there is a need to find a bioreactor that can be adapted to improve antigen production.
Disclosure of Invention
The invention aims to provide a bioreactor for improving the antigen yield, which aims to solve the problems that in the prior art, in order to obtain good oxygen and mass transfer in antigen cell culture, the stirring efficiency can be effectively improved, and the cell culture efficiency of the bioreactor is improved; the culture technology can be used for producing antigen vaccines of the avian influenza virus.
The technical scheme adopted by the invention is as follows: the invention relates to a bioreactor for improving antigen yield, which comprises: the device comprises a closed tank-shaped shell, a stirring device, a progressive spiral pipe for stirring, a rotating shaft, a storage device, an inverted triangular shell of the storage device, a flow guide screen plate, a fluid circulating pipeline, a fluid circulating pump, a deposition box body at the bottom of the progressive spiral pipe and a liquid level sensor.
The liquid level sensor is used for displaying the liquid level height in the bioreactor in real time, and is convenient for real-time control of the culture process of antigen cells in the bioreactor.
Wherein, the rotating shaft penetrates through the axis of the shell in a rotating and sealing way, and the storage device is fixedly connected to the upper part of the shell and is still relative to the shell; the rotating shaft penetrates through the center of the storage device in a rotating and sealing manner, and the bottom of the storage device is connected with a stirring device; the bottom of the rotating shaft is fixedly connected with the top of the stirring device and rotates together with the stirring device, and the bottom of the storage device and the top of the stirring device are kept in rotary sealing while the rotating shaft rotates.
Wherein, a circulating pipeline is connected between the inlet of the storage device and the bottom of the shell, and a liquid circulating pump is connected in series on the circulating pipeline; the storage device comprises an inverted triangular shell, and can effectively ensure that the internal fluid flows downwards smoothly; the lower part of the storage device is positioned on the upper part of the progressive spiral pipe, and the flow guide screen plate is welded on the storage device and plays roles of filtering and guiding flow.
The stirring device comprises a progressive spiral pipe for stirring and a deposition box body positioned at the bottom of the progressive spiral pipe, and the deposition box body is communicated with a progressive spiral pipe pipeline; a plurality of small holes are formed on the outer edge side of the progressive spiral pipe around the rotating shaft; the small holes are uniformly distributed along the circumference of the progressive spiral pipe 3, so that the flow of the fluid flowing into the bioreactor along the circumference is more uniform; fluid in the storage device can enter the storage device through a circulating pipeline, then flows into the progressive spiral tube of the stirring device through the flow guide screen plate, drives the progressive spiral tube to rotate through the rotating shaft, and further enters the bioreactor through the small holes to complete cell culture.
Wherein the diameter of the top of the progressive spiral pipe is a, and the diameter of the bottom of the progressive spiral pipe is b; wherein a is smaller than b, thus forming a progressive spiral stirring device with gradually increasing diameter from top to bottom; according to the invention, according to the change of the cell density in the bioreactor, a progressive spiral tube in a spiral shape is adopted from top to bottom, the distribution of the cell density in the bioreactor and the timely structural change of a stirring device are combined, and the stirring efficiency of the stirring device in the bioreactor for high-density large-scale cell culture is effectively improved.
Wherein, the deposit box is located the bottom of progressive spiral pipe, and the progressive spiral pipe of pipe connection for collect the fluid precipitate in storage device and the progressive spiral pipe, can the effectual purity that guarantees fluid, and the convenience is to bioreactor's cleanness after the cell finishes.
The top of the progressive spiral pipe is in rotary sealing connection with the bottom of the storage device, the rotating shaft is in sealing connection with the top of the shell, and the rotating shaft is in sealing connection with the top of the storage device; all adopt rotary joint seal.
The rotating shaft is fixedly connected with the top of the progressive spiral tube at the bottom of the storage device, and the rotating shaft and the progressive spiral tube can be fixedly welded and connected in a symmetrical mode; the rotating shaft and the progressive spiral tube can be manufactured in an integrated forming mode.
The invention has the advantages that: for the bioreactor for improving the antigen yield, the internal structure of the bioreactor is improved, and the culture solution is forcibly circulated by using a circulating pump paddle of a circulating pipeline, so that the contact area between cells and fluid is greatly increased; the forced circulation results in enhanced liquid flowability and excellent mass transfer effect.
Drawings
FIG. 1 is a schematic overall cross-sectional structural view of the present invention;
FIG. 2 is a schematic view of the connection of the shaft of the present invention to the progressive spiral tube, i.e., a sectional view A-A in FIG. 1;
icon: 1-a closed can-like housing; 2-a stirring device; 3-a progressive spiral pipe; 4-a rotating shaft; 5-a storage device; 6-inverted triangular shell; 7-a flow guiding screen plate; 8-a circulation line; 9-a fluid circulation pump; 10-deposition chamber.
Detailed Description
Referring to fig. 1, the present invention relates to a bioreactor for increasing antigen production, comprising: the device comprises a closed tank-shaped shell 1, a stirring device 2, a progressive spiral pipe 3 for stirring, a rotating shaft 4, a storage device 5, an inverted triangular shell 6 of the storage device 5, a flow guide screen plate 7, a fluid circulation pipeline 8, a fluid circulation pump 9, a deposition box body 10 at the bottom of the progressive spiral pipe 3 and a liquid level sensor 11.
Wherein, the liquid level sensor 11 is used for displaying the liquid level height in the bioreactor in real time, which is convenient for the real-time control of the culture process of the antigen cells in the bioreactor.
Wherein, the rotating shaft 4 penetrates through the axis of the shell 1 in a rotating and sealing way, and the storage device 5 is fixedly connected to the upper part of the shell 1 and is still relative to the shell 1; the rotating shaft 4 penetrates through the center of the storage device 5 in a rotating and sealing manner, and the bottom of the storage device 5 is connected with the stirring device 2; the bottom of the rotating shaft 4 is fixedly connected with the top of the stirring device 2 and rotates together with the stirring device, and the bottom of the storage device 5 keeps rotating and sealing with the top of the stirring device 2.
Wherein, a circulating pipeline 8 is connected between the inlet of the storage device 5 and the bottom of the shell 1, and a liquid circulating pump 9 is connected in series on the circulating pipeline 8; the storage device 5 comprises an inverted triangular shell 6, which can effectively ensure that the internal fluid flows downwards smoothly; the lower part of the storage device 5 and the upper part of the progressive spiral pipe 3 are provided with a flow guide screen plate 7, and the flow guide screen plate 7 is welded on the storage device 5 to play roles of filtering and flow guiding.
The stirring device 2 comprises a progressive spiral pipe 3 for stirring and a deposition box body 10 positioned at the bottom of the progressive spiral pipe 3, and the deposition box body 10 is communicated with the progressive spiral pipe 3 through a pipeline; a plurality of small holes 51 are arranged around the rotating shaft 4 and at the outer edge side of the progressive spiral tube 3; the small holes 51 are uniformly distributed along the circumference of the progressive spiral tube 3 to make the flow of fluid circumferentially flowing into the bioreactor more uniform; the fluid in the storage device 5 can enter the storage device 5 through the circulating pipeline 8, then flows into the progressive spiral tube 3 of the stirring device 2 through the diversion net plate 7, drives the progressive spiral tube 3 to rotate through the rotating shaft 4, and further enters the bioreactor through the small holes 51 to complete the culture of cells.
Wherein the diameter of the top of the progressive spiral tube 3 is a, and the diameter of the bottom is b; wherein a is smaller than b, thus forming a progressive spiral stirring device with gradually increasing diameter from top to bottom; in the bioreactor for high-density large-scale cell culture, the cell density at the lower part is gradually higher than that at the upper part, according to the change of the cell density in the bioreactor, the progressive spiral tube 3 in the spiral shape is adopted in a progressive manner from top to bottom, the distribution of the cell density in the bioreactor and the timely structural change of the stirring device are combined, and the stirring efficiency of the stirring device in the bioreactor for high-density large-scale cell culture is effectively improved.
Wherein, deposit box 10 and be located progressive spiral pipe 3's bottom, progressive spiral pipe 3 is connected to the pipeline for collect the fluid precipitate in storage device 5 and the progressive spiral pipe 3, can effectually guarantee fluidic purity, and the convenience is to bioreactor's cleanness after the cell finishes.
Wherein, the top of the progressive spiral tube 3 is in rotary sealing connection with the bottom of the storage device 5, the rotating shaft 4 is in sealing connection with the top of the shell 1, and the rotating shaft 4 is in sealing connection with the top of the storage device 5; all adopt rotary joint seal.
Referring to fig. 2, at the bottom of the storage device 5, the rotating shaft 4 is fixedly connected with the top of the progressive spiral tube 3, and the rotating shaft 4 and the progressive spiral tube 3 can be fixedly welded in a symmetrical manner; the rotating shaft 4 and the progressive spiral tube 3 can be manufactured in an integrated forming mode.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (8)
1. A bioreactor for increasing antigen production, comprising: the device comprises a closed tank-shaped shell (1), a stirring device (2), a progressive spiral pipe (3) for stirring, a rotating shaft (4), a storage device (5), an inverted triangular shell (6) of the storage device (5), a flow guide screen plate (7), a fluid circulating pipeline (8), a fluid circulating pump (9), a deposition box body (10) at the bottom of the progressive spiral pipe (3) and a liquid level sensor (11);
wherein, the rotating shaft (4) penetrates through the axis of the shell (1) in a rotating and sealing way, and the storage device (5) is fixedly connected to the upper part of the shell (1) and is still relative to the shell (1); the rotating shaft (4) penetrates through the center of the storage device (5) in a rotating and sealing manner, and the bottom of the storage device (5) is connected with the stirring device (2); the bottom of the rotating shaft (4) is fixedly connected with the top of the stirring device (2) and rotates together with the stirring device, and the bottom of the storage device (5) and the top of the stirring device (2) keep rotating and sealing while rotating;
wherein, a circulating pipeline (8) is connected between the inlet of the storage device (5) and the bottom of the shell (1), and a liquid circulating pump (9) is connected in series on the circulating pipeline (8); the storage device (5) comprises an inverted triangular shell (6); the lower part of the storage device (5) is positioned at the upper part of the progressive spiral pipe (3) and is provided with a flow guide screen plate (7), and the flow guide screen plate (7) is welded on the storage device (5);
the stirring device (2) comprises a progressive spiral pipe (3) for stirring and a deposition box body (10) positioned at the bottom of the progressive spiral pipe (3), and the deposition box body (10) is communicated with the progressive spiral pipe (3) through a pipeline;
wherein the deposition box body (10) is positioned at the bottom of the progressive spiral pipe (3), and a pipeline is connected with the progressive spiral pipe (3) and is used for collecting fluid sediments in the storage device (5) and the progressive spiral pipe (3);
a plurality of small holes (51) are arranged around the rotating shaft (4) and on the outer edge side of the progressive spiral pipe (3); the small holes (51) are uniformly distributed along the circumference of the progressive spiral tube (3); fluid in the storage device (5) can enter the storage device (5) through a circulating pipeline (8), then flows into the progressive spiral tube (3) of the stirring device (2) through a flow guide screen plate (7), the progressive spiral tube (3) is driven to rotate through a rotating shaft (4), and the fluid further enters the bioreactor through the small holes (51) to complete the culture of cells;
at the bottom of the storage device (5), the rotating shaft (4) is fixedly connected with the top of the progressive spiral tube (3), and the rotating shaft (4) and the progressive spiral tube (3) are fixedly welded in a symmetrical mode;
it is characterized in that the preparation method is characterized in that,
the top diameter of the progressive spiral tube (3) is a, and the bottom diameter is b; wherein a is smaller than b, and a progressive spiral stirring device with gradually increasing diameter from top to bottom is formed.
2. Bioreactor for increasing the yield of antigens according to claim 1, characterized in that the top of the progressive spiral tube (3) is in rotary sealed connection with the bottom of the storage device (5), the rotating shaft (4) is in sealed connection with the top of the housing (1), the rotating shaft (4) is in sealed connection with the top of the storage device (5); all adopt rotary joint seal.
3. Bioreactor for increasing the production of antigens according to claim 1, characterized in that the top of the progressive spiral tube (3) is sealed with a rotary joint in a rotary sealing connection with the bottom of the storage means (5).
4. Bioreactor for increasing the yield of antigens according to claim 1, characterized in that the sealing connection of the rotating shaft (4) to the top of the housing (1) is sealed by means of a rotating joint.
5. Bioreactor for increasing the yield of antigens according to claim 1, characterized in that the sealing connection of the rotating shaft (4) to the top of the storage device (5) is sealed by a rotating joint.
6. Bioreactor for increasing the yield of antigens according to claim 1, characterized in that the top of the progressive spiral tube (3) is in rotary sealed connection with the bottom of the storage device (5), the rotating shaft (4) is in sealed connection with the top of the housing (1), the rotating shaft (4) is in sealed connection with the top of the storage device (5); all adopt rotary joint seal.
7. The bioreactor for increasing antigen production according to claim 1, wherein the liquid level sensor 11 is used for displaying the liquid level in the bioreactor in real time.
8. Bioreactor for increasing the yield of antigens according to claim 8, characterized in that the rotating shaft (4) and the progressive spiral tube (3) are made in an integrated manner.
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| CN202010582982.5A CN111676136B (en) | 2020-06-23 | 2020-06-23 | Bioreactor for improving antigen yield |
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| CN202010582982.5A CN111676136B (en) | 2020-06-23 | 2020-06-23 | Bioreactor for improving antigen yield |
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