CN215975856U - Perfusion culture device with single magnetic pump for alternative tangential flow filtration - Google Patents
Perfusion culture device with single magnetic pump for alternative tangential flow filtration Download PDFInfo
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- CN215975856U CN215975856U CN202122260570.2U CN202122260570U CN215975856U CN 215975856 U CN215975856 U CN 215975856U CN 202122260570 U CN202122260570 U CN 202122260570U CN 215975856 U CN215975856 U CN 215975856U
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Abstract
The utility model discloses a perfusion culture device with a single magnetic pump for alternative tangential flow filtration, which comprises a bioreactor, wherein the side wall of the lower part of the bioreactor is provided with a material outlet and a material backflow port, and the material outlet is connected with the magnetic pump through a hose; the magnetic pump is connected with the filtering device through a parallel pipeline; the parallel pipeline is connected with a hose and a material reflux port; when the filter device is used, one end of the first connecting port and one end of the second connecting port of the filter device are a material inlet, the other end of the first connecting port and the other end of the second connecting port are a backflow port, and the third connecting port of the filter device is a permeate outlet; a peristaltic pump is arranged between the permeate collection tank and the filtering device. According to the utility model, the direction of material flowing at the upstream of the filter can be changed by alternately opening the four valves which are connected with the filter in parallel in pairs, so that an alternate tangential flow filtration mode is formed, the driving of a plurality of magnetic pumps in different directions is not required, the same effect can be achieved by using a single magnetic pump, the structure is simple, the operation is convenient, and the cost is low.
Description
Technical Field
The utility model relates to a perfusion culture device, in particular to a perfusion culture device adopting a single magnetic pump to alternately filter tangential flow.
Background
At present, batch culture and fed-batch culture are most widely applied, and perfusion culture can produce products with higher quality and accumulate higher cell concentration than batch fed-batch culture. Perfusion culture techniques are still in the laboratory or pilot plant stage and have not been applied to production on a large scale. The perfusion culture technique is a method of increasing the discharge of harmful metabolites or products on the basis of fed-batch culture. The cell culture mode of supplementing substrates required by cell growth, discharging products produced after cell metabolism or substances harmful to cell growth, and trapping cells in a reactor realizes high-density cell culture and high product yield. Commonly used perfusion culture techniques are: inclined sedimentation, sonic sedimentation, centrifugation, vortex separation, rotary filtration, wave filtration, Tangential Flow Filtration (TFF). In tangential flow filtration, a pump pushes fluid across the surface of the filter membrane, scouring away molecules trapped thereon, thereby minimizing fouling of the membrane surface.
The perfusion culture method firstly pays attention to sterility guarantee and rejection rate, the cell culture process is a sterile culture process, and long-time cell culture can be realized only by guaranteeing sterility. The retention rate is not high and the aim of accumulating high cell density is not achieved. The sterility of the inclined sedimentation is difficult to guarantee, the retention rate of the acoustic sedimentation, the rotary filtration and the vortex separation is not high, and the large-scale application of the inclined sedimentation in perfusion culture is limited. Secondly, shear force, too much shear force can damage cells and affect the activity of the cells. The excessive shearing force of centrifugation also limits the application of the centrifugal force in perfusion culture. There is also concern over long term operation, with wave filtration being less capable. The most promising perfusion culture techniques are Tangential Flow Filtration (TFF) and alternating tangential flow filtration (ATF). Both compare that TFF is more limited than ATF. Because TFF has higher shear than ATF, and accumulates lower cell concentration and cell viability, TFF is much poorer in sterility and long-term operation capability than ATF.
The TFF technology can cause the blockage of the filter membrane in the process of long-time operation, and the long-time operation can cause the blockage of the filter membrane and cannot be used continuously because the upstream of the material filter membrane is in one-way flow.
Therefore, the development of a perfusion culture device with a single magnetic pump and alternating tangential flow filtration is a problem to be solved by those skilled in the art.
SUMMERY OF THE UTILITY MODEL
The utility model aims to solve the defects and provides a perfusion culture device with a single magnetic pump for alternative tangential flow filtration.
The utility model adopts a connection mode that four valves are connected in parallel with the filter in pairs, and skillfully enables a single unidirectional magnetic pump to play the effect of the alternate flow of materials at the upstream of the filtering membrane. Compared with a rotor pump or a quaternary diaphragm pump, the magnetic pump can obviously reduce the shearing force on materials. The process that the materials flow alternately through the valve is equivalent to self-cleaning the surface of the filtering membrane, so that the blockage of the membrane is avoided, and the membrane can run for a long time. The alternate tangential flow filtration technology of the utility model can be well applied to concentration perfusion, is used for producing protein products, can realize continuous concentration perfusion, can realize continuous harvest, is convenient for downstream purification, and can reach extremely high cell concentration. Can also be applied to concentrated fed-batch culture, seed bank construction, rapid production and the like.
The above object of the present invention is achieved by the following technical means: a perfusion culture device with a single magnetic pump for alternative tangential flow filtration comprises a bioreactor, a magnetic pump, a filter device, a permeate collection tank, a peristaltic pump and parallel pipelines,
a material outlet and a material reflux port are formed in the side wall of the lower part of the bioreactor, and the material outlet is connected with the magnetic pump through a hose; the magnetic pump is connected with the filtering device through a parallel pipeline; the parallel pipeline is connected to the material backflow port through a hose;
when the filter device is used, one end of the first connecting port and one end of the second connecting port of the filter device are a material inlet, the other end of the first connecting port and the other end of the second connecting port are a backflow port, and the third connecting port of the filter device is a permeate outlet and is connected to a permeate collecting tank through a hose;
a peristaltic pump is arranged between the permeate collection tank and the filtering device.
Furthermore, the parallel pipeline comprises a valve group A and a valve group B, the valve group A, the valve group B and the filtering device are connected in parallel, and an access port of the parallel pipeline is positioned between the first valve and the third valve and is connected with the magnetic pump; and the return port of the parallel pipeline is positioned between the second valve and the fourth valve and is connected into the material return port.
Further, the valve group a includes a first valve and a third valve, and the first valve and the third valve are connected in series.
Further, the valve group B comprises a second valve and a fourth valve, and the second valve and the fourth valve are connected in series.
Further, an ultrasonic flowmeter is arranged between the magnetic pump and the parallel pipeline.
Further, the ultrasonic flowmeter is sandwiched between the outer walls of the hoses, and measures the flow rate of the liquid in the hoses by transmitting a signal to the hoses, transmitting the signal through the hoses, and detecting the transmitted signal on the other side.
Furthermore, the hose and the magnetic pump or the filtering device are connected in a quick-connection type clamp connection mode.
Furthermore, the connection mode of the hose and the bioreactor or the permeate receiving tank can be a quick-connection type clamp connection, and can also be a sanitary connection mode such as an aseptic flange.
Further, an ultrasonic flow is arranged between the permeate outlet of the filtering device and the permeate collecting tank and used for measuring and monitoring the flow meter.
The working principle of the utility model is as follows:
after the magnetic pump is started, the materials in the bioreactor begin to enter the hose from the material outlet and flow to the parallel pipeline from the material outlet along the transmission direction of the magnetic pump.
The key for realizing the alternation of the upstream material flow direction of the filtering device lies in the regulation and control of four valves of the parallel pipeline.
When the first valve and the fourth valve are kept open and the second valve and the third valve are closed, the material enters the filtering device from the first connecting port through the pipeline where the first valve is located, flows out from the second connecting port, and flows back to the bioreactor 1 after passing through the fourth valve;
when the second valve and the third valve are kept open and the first valve and the fourth valve are closed, the material enters the filtering device through the pipeline where the third valve is located and the second connecting port, flows out of the first connecting port, and flows back to the bioreactor after passing through the second valve.
From the above working principle, it can be seen that the two different valve opening modes cause the material to enter the filter in different directions, but for the bioreactor, the material outflow and inflow directions are always the same.
In the process, the peristaltic pump is always kept on, and the permeate can be enabled to flow to the permeate collection tank through the control of the peristaltic pump regardless of the flow direction of the upstream materials.
Further, in the present invention:
1. a bioreactor: the bioreactor is a general term for a device for performing liquid culture of mammalian cells.
2. A magnetic pump: the magnetic-driven type material-sealing device has the greatest characteristic that the magnetic-driven type material-sealing device is driven by magnetic force, does not need mechanical sealing, and thoroughly solves the risk of material leakage or pollution; the impeller of the magnetic pump is a specially designed impeller with low shearing force, and is very suitable for the biopharmaceutical industry. The shear force of the cells or protein product is very low.
3. An ultrasonic flowmeter: an ultrasonic flowmeter is a meter which transmits ultrasonic waves through a hose and detects changes in the transmitted signals on the opposite side of the hose to reflect the flow rate in the hose. The flowmeter is not in direct contact with the material and is clamped outside the hose.
4. A filtering device: such as hollow membrane filtration, the upstream end of the membrane flows up and down and the permeate end can flow to a permeate collection tank. The upper and lower interfaces of the filtering device are a liquid inlet and a reflux port respectively, and the side wall port is a liquid permeation port.
5. A permeate collection tank: liquid at the permeable end of the filter is collected to the permeate collection tank through the peristaltic pump, so that cell metabolites or metabolic waste in the bioreactor are discharged during perfusion culture.
Compared with the prior art, the utility model has the advantages that:
1. through the mode that four valves parallelly connected with two liang of filters are opened in turn, can change the material in the direction of filter upper reaches flow, form the tangential flow filtration mode in turn, do not need the different magnetic drive pump drive of a plurality of directions, use single magnetic drive pump just can reach same effect, simple structure convenient operation, it is with low costs.
2. The liquid material flows alternately in the filtering device, so that the problem that the filtering membrane is easy to stack is solved, the liquid material flows continuously and alternately, alternate and reciprocating scouring is formed on the surface of the filtering membrane, and the filtering membrane can be in a good filtering state for a long time.
3. The filter sees through the end and connects the peristaltic pump, can independently control the collection speed of permeate liquid, greatly increased the permeation efficiency of filter, reduces the jam probability of filter membrane, improves whole production efficiency.
3. Guarantee of sterile interface: the magnetic pump is driven by magnetic force, and the stirring impeller and the motor are completely isolated, so that the liquid leakage and pollution risks can be avoided, and the magnetic pump can operate for a long time. And the pump head, the filtering device and the hose of the magnetic pump can be connected with the bioreactor or the permeate collecting device after off-line sterilization. The aseptic connection scheme to the bioreactor or permeate collection tank may be aseptic docking using a tube cutter.
4. At present, the device is connected by a hose, sterilized off-line and used continuously after being in aseptic butt joint. According to the production scale, the device can also be designed to be connected with a stainless steel hard pipeline, the magnetic pump and the filtering device are both designed to be stainless steel shells, the online sterilization function of the device is realized, manual operation is not needed, and full-automatic cleaning, sterilization and liquid material alternate filtration are realized.
Drawings
Fig. 1 is a schematic structural view of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
As shown in figure 1, the perfusion culture device with single magnetic pump alternating tangential flow filtration comprises a bioreactor 1, a magnetic pump 3, a filter device 5, a permeate collection tank 6, a peristaltic pump 7 and a parallel pipeline 9,
a material outlet 1-1 and a material reflux port 1-2 are formed in the side wall of the lower part of the bioreactor 1, and the material outlet 1-1 is connected with a magnetic pump 3 through a hose 8; the magnetic pump 3 is connected with the filtering device 5 through a parallel pipeline 9; the parallel pipeline 9 is connected to the material return port 1-2 through a hose 10;
when in use, one end of the first connecting port 5-1 and the second connecting port 5-2 of the filtering device 5 is a material inlet, the other end of the first connecting port is a reflux port, and the third connecting port 5-3 of the filtering device is a permeate outlet and is connected to a permeate collecting tank 6 by a hose;
a peristaltic pump 7 is arranged between the permeate collection tank 6 and the filtering device 5.
Further, the parallel pipeline 9 comprises a valve group A and a valve group B, the valve group A, the valve group B and the filtering device 5 are connected in parallel, and an access port of the parallel pipeline 9 is positioned between the first valve 2-1 and the third valve 2-3 and is connected with the magnetic pump 3; and the return port of the parallel pipeline 9 is positioned between the second valve 2-2 and the fourth valve 2-4 and is connected into the material return port 1-2.
Further, the valve group A comprises a first valve 2-1 and a third valve 2-3, and the first valve 2-1 and the third valve 2-3 are connected in series.
Further, the valve group B comprises a second valve 2-2 and a fourth valve 2-4, and the second valve 2-2 and the fourth valve 2-4 are connected in series.
Further, an ultrasonic flowmeter 4 is installed between the magnetic pump 3 and the parallel pipeline 9.
Further, the ultrasonic flowmeter 4 is sandwiched between the outer walls of the tubes, and measures the flow rate of the liquid in the tubes by transmitting a signal to the tubes, transmitting the signal through the tubes, and detecting the transmitted signal on the other side.
Furthermore, the hose and the magnetic pump 3 or the filtering device 5 are connected in a quick-connection type clamp connection mode.
Furthermore, the connection mode of the hose and the bioreactor or the permeate receiving tank can be a quick-connection type clamp connection, and can also be a sanitary connection mode such as an aseptic flange.
Further, an ultrasonic flow meter is installed between the permeate outlet of the filtering device 5 and the permeate collection tank 6 for monitoring the flow rate. This embodiment is equipped with only one ultrasonic flow meter.
The working principle of the utility model is as follows:
after the magnetic pump 3 is started, the materials in the bioreactor 1 start to enter the hose 8 from the material outlet 1-1, and flow from the material outlet 1-1 to the parallel pipeline 9 along the transmission direction of the magnetic pump 3.
The key for realizing the alternation of the upstream material flow direction of the filtering device lies in the regulation and control of four valves of the parallel pipeline.
When the first valve 2-1 and the fourth valve 2-4 are kept open and the second valve 2-2 and the third valve 2-3 are kept closed, the material enters the filtering device from the first connecting port 5-1 through the pipeline where the first valve 2-1 is located, flows out from the second connecting port 5-2, flows back to the bioreactor 1 after passing through the fourth valve 2-4;
when the second valve 2-2 and the third valve 2-3 are kept open and the first valve 2-1 and the fourth valve 2-4 are closed, the material enters the filtering device through the pipeline where the third valve 2-3 is located and the second connecting port 5-2, flows out of the first connecting port 5-1, flows back to the bioreactor 1 after passing through the second valve 2-2.
From the above working principle, it can be seen that the two different valve opening modes cause the material to enter the filter in different directions, but for the bioreactor, the material outflow and inflow directions are always the same.
During the above process, the peristaltic pump 7 is kept on all the time, and the permeate can be flowed to the permeate collection tank by the control of the peristaltic pump 7 regardless of the flow direction of the upstream material.
Further, in the present invention:
1. a bioreactor: the bioreactor is a general term for a device for performing liquid culture of mammalian cells.
2. A magnetic pump: the magnetic-driven type material-sealing device has the greatest characteristic that the magnetic-driven type material-sealing device is driven by magnetic force, does not need mechanical sealing, and thoroughly solves the risk of material leakage or pollution; the impeller of the magnetic pump is a specially designed impeller with low shearing force, and is very suitable for the biopharmaceutical industry. The shear force of the cells or protein product is very low.
3. An ultrasonic flowmeter: an ultrasonic flowmeter is a meter which transmits ultrasonic waves through a hose and detects changes in the transmitted signals on the opposite side of the hose to reflect the flow rate in the hose. The flowmeter is not in direct contact with the material and is clamped outside the hose.
4. A filtering device: such as hollow membrane filtration, the upstream end of the membrane flows up and down and the permeate end can flow to a permeate collection tank. The upper and lower interfaces of the filtering device are a liquid inlet and a reflux port respectively, and the side wall port is a liquid permeation port.
5. A permeate collection tank: liquid at the permeable end of the filter is collected to the permeate collection tank through the peristaltic pump, so that cell metabolites or metabolic waste in the bioreactor are discharged during perfusion culture.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes performed by the present specification and drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (9)
1. The utility model provides a perfusion culture apparatus of single magnetic pump alternative tangential flow filtration which characterized in that: comprises a bioreactor, a magnetic pump, a filtering device, a permeate collecting tank, a peristaltic pump and parallel pipelines,
a material outlet and a material reflux port are formed in the side wall of the lower part of the bioreactor, and the material outlet is connected with the magnetic pump through a hose; the magnetic pump is connected with the filtering device through a parallel pipeline; the parallel pipeline is connected with a hose and a material reflux port;
when the filter device is used, one end of the first connecting port and one end of the second connecting port of the filter device are a material inlet, the other end of the first connecting port and the other end of the second connecting port are a backflow port, and the third connecting port of the filter device is a permeate outlet and is connected to a permeate collecting tank through a hose;
a peristaltic pump is arranged between the permeate collection tank and the filtering device.
2. The perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 1, wherein: the parallel pipeline comprises a valve group A and a valve group B, the valve group A, the valve group B and the filtering device are connected in parallel, and an access port of the parallel pipeline is positioned between the first valve and the third valve and is connected with the magnetic pump; and the return port of the parallel pipeline is positioned between the second valve and the fourth valve and is connected into the material return port.
3. The perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 2, wherein: the valve group A comprises a first valve and a third valve which are connected in series.
4. The perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 2, wherein: the valve group B comprises a second valve and a fourth valve, and the second valve and the fourth valve are connected in series.
5. The perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 1, wherein: an ultrasonic flowmeter is arranged between the magnetic pump and the parallel pipeline.
6. A perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 5, wherein: the ultrasonic flowmeter is clamped on the outer wall of the hose, transmits a signal to the hose, and detects the transmitted signal on the other side after transmitting the signal through the hose to measure the flow rate of the liquid in the hose.
7. The perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 1, wherein: the hose and the magnetic pump or the filtering device are connected in a quick connection type clamping connection mode.
8. The perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 1, wherein: the connection mode of the hose and the bioreactor or the permeate receiving tank is quick connection type clamp connection or aseptic flange connection.
9. The perfusion culture device with single magnetic pump alternating tangential flow filtration according to claim 1, wherein: an ultrasonic flowmeter is arranged between the permeate outlet of the filtering device and the permeate collecting tank.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202122260570.2U CN215975856U (en) | 2021-09-17 | 2021-09-17 | Perfusion culture device with single magnetic pump for alternative tangential flow filtration |
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| CN202122260570.2U CN215975856U (en) | 2021-09-17 | 2021-09-17 | Perfusion culture device with single magnetic pump for alternative tangential flow filtration |
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| CN215975856U true CN215975856U (en) | 2022-03-08 |
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| CN202122260570.2U Active CN215975856U (en) | 2021-09-17 | 2021-09-17 | Perfusion culture device with single magnetic pump for alternative tangential flow filtration |
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