CN111154647A - Closed circulation external exchange biological product reactor - Google Patents
Closed circulation external exchange biological product reactor Download PDFInfo
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Abstract
The invention discloses a closed circulation external exchange biological product reactor which comprises a bioreactor body, a main circulation pipeline, a main circulation channel peristaltic pump, a liquid level sensor, a gas escape channel and filter, a culture medium injection three-way pipeline, a gas exchange module, an O2/N2 channel, a dissolved oxygen and PH sensor group, a culture medium valve, a culture medium pump body, a culture medium storage unit, an ATF (automatic transfer function) matching module, a sterile sample acquisition channel, automatic sample transfer and an online detection equipment group. The cells and the production products are ensured to be uniformly distributed in the reactor; maintaining the excellent growth state of the cells; the damage of the shearing force to the cells in the production process can be effectively avoided, and the excellent growth state of the cells is ensured; the generation of bubbles in the production process is greatly reduced, and the pollution occurrence probability is reduced; meanwhile, the influence of the impact force generated by the burst of the bubbles in the fluid on the cells is avoided, and an ideal environment is provided for the growth of the cells.
Description
Technical Field
The invention belongs to the technical field of external exchange biological product reactors, and particularly relates to a closed circulation external exchange biological product reactor.
Background
Along with the rapid development of immunotherapy, the demands for allogeneic immune cell therapy, cell therapy virus production and customized antibody production on human/animal modified cell production are rapidly increased. The traditional manual production mode is based on culture bottles, shake flasks and culture bags, and the production scale is difficult to scale up. The existing large-scale production of biological medicines mainly depends on a bioreactor. The existing mainstream stirring bioreactor can achieve a large production scale, but needs a stirring paddle to realize the circulation of nutrient substances, dissolved gas and the like in a culture system. The human source and animal improved cells have no cell walls, are not domesticated, are sensitive to shearing force and impact force generated by a stirring paddle, and are not effectively solved by cell damage and apoptosis, so that the human source/animal improved suspended cells cannot be effectively produced in a large scale in the traditional stirring type reactor. Other types of reactors, such as the rocking bed reactor, also have problems with cell shear forces when scaling up, and culture densities that are difficult to exceed on the order of 1Ex7/ml, and throughput that is difficult to meet market demands. There is a need for a bioreactor for large-scale production of human/animal suspension cells to meet the requirements of efficient, large-scale, and low-cost production.
Disclosure of Invention
The object of the present invention is to provide: a closed circulation external exchange biological product reactor provides a high-efficiency bioreactor and a matching system suitable for single-batch production of a large number of (xenogenous) animal suspension cells, humanized suspension cells, viruses and antibodies. Filling the market vacancy; the problems of shearing force and impact force caused by a stirring paddle of the traditional reactor are solved, and the problems of cell damage and cell apoptosis caused by the traditional reactor in the process of producing animal/human cells are avoided; compared with the existing production mode, the single-time production yield efficiency is greatly improved. Under the condition of equal cost input, the yield is improved by one order of magnitude to reach the actual yield requirement. Greatly improving the exchange flux and exchange efficiency of nutrients; realizing multi-parameter continuous data acquisition and main biochemical control index closed-loop control of the reactor. Effective data mining in the production process is realized, and the process is continuously optimized on line, so that the problems in the background technology are solved.
In order to achieve the purpose, the invention provides the following technical scheme: a closed circulation external exchange biological product reactor comprises a bioreactor body, a main circulation pipeline, a main circulation channel peristaltic pump, a liquid level sensor, a gas escape channel, a filter, a culture medium injection three-way pipeline, a gas exchange module, an O2/N2 channel, a dissolved oxygen and PH sensor group, a culture medium valve, a culture medium pump body, a culture medium storage unit, an ATF (automatic transfer function) matching module, an aseptic sample collection channel, automatic sample transfer and an online detection equipment group.
Further, the corresponding functions are as follows:
the bioreactor body: the major sites for cell growth and the continuous maintenance of cells in suspension. The nutrient substances after gas exchange and culture medium injection are effectively diffused in the reactor;
a main circulation pipeline: the solution in the circulating main reactor is subjected to dissolved oxygen and carbon dioxide discharge exchange in a gas exchange module, and a new culture medium is injected into a culture solution tee joint in an exchange way;
main circulation channel peristaltic pump: used for driving the solution in the bioreactor to circulate among the main circulation pipeline, the gas exchanger and the culture medium adding channel;
a liquid level sensor: the device is used for continuously detecting the height of the liquid level in the main reactor and is used for closed-loop control of the liquid amount of used culture medium discharge and new culture medium addition;
gas escape channels and filters: during the production process, the pressure in the reactor rises due to the escape of dissolved gases and the volatilization of liquids. The channel discharges the excessive gas;
injecting a culture medium into the three-way pipeline: when the culture medium is added, the culture medium pumping pipeline and the main circulation pipeline are communicated, new culture medium enters the reactor main body, and when the tee joint is switched, the culture medium does not enter the main circulation pipeline;
a gas exchange module: the special membrane type gas exchange unit is used for carrying out efficient gas exchange. When the liquid passes through, O2 provided by a special pipeline is efficiently dissolved into the culture solution, and simultaneously, excessive dissolved CO2 in the culture solution is discharged through the introduction and dissolution of N2, and the pH value of the culture solution is adjusted through gas exchange;
O2/N2 channel: for supplying the sterilized gas subjected to the strict filtration treatment to the gas exchanger for culture solution dissolved gas exchange;
dissolved oxygen and PH sensor set: by measuring the dissolved oxygen concentration and the PH value before/after gas exchange, dynamically adjusting the liquid flow of the main circulation passing through the gas exchanger, the proportion of O2/N2 and the pressure of O2/N2, and realizing the dynamic adjustment of the dissolved oxygen concentration and the PH value based on target set values;
culture medium valve: connecting or closing the connection of the culture medium and the main circulation pipeline;
culture medium pump body: pumping the culture medium into the main circulation channel;
a medium storage unit: storing the culture medium for mass production for a long time in a storage environment of 4 ℃ degrees;
ATF enrichment unit: in the production process, continuously or periodically discharging the used culture medium, filtering the enriched cells, and returning the cells enriched in the module to the culture system again according to the requirement;
ATF matching module: used culture medium used for collecting the ATF is discharged to a waste liquid collecting barrel, and the ATF module is isolated from an external channel when the ATF module is not in work;
sterile sample collection channel: a culture solution sample in the reactor body is regularly and aseptically collected and is used for comprehensive index detection and culture liquid online adjustment, and the continuous control of main indexes in the production process is realized;
automatic sample transfer: after aseptically sampling to a sample tube, transferring the sample to each detection device on a test bench;
an online detection device group: the method is used for online production index detection and realizes closed-loop control of main biochemical indexes.
Further, the pH values of the dissolved oxygen and the pH sensor group are absolute values and difference values, and the liquid flow rate is the liquid rate; the dissolved oxygen and PH sensor group comprises a concentration detection unit before gas exchange and a concentration detection unit after gas exchange; the ATF matching module comprises an ATF valve, an ATF pump body, an ATF pipeline and an ATF waste liquid collecting unit; the sterile sample collecting channel comprises a sampling joint and a sampling valve; the online detection equipment group comprises a cell counting module and a biochemical index detection module.
Further, the dissolved gas concentration adjusting step in the production process is as follows:
s1: first, DO measurement before gas exchange;
s2: secondly, measuring the PH before gas exchange;
s3: measuring DO after gas exchange;
s4: measuring the PH after gas exchange;
s5: obtaining the difference value of DO and PH before and after gas exchange;
s6: calculating O2、N2Exchange efficiency target of (2);
s7: calculating the flow rate of the culture liquid in the gas exchanger;
s8: calculating O2、N2Flow rate in the pipeline;
s9: controlling and adjusting the flow of the culture liquid; control adjustment O2/N2And (4) flow rate.
Further, the steps S1 to S9 are repeated, and the detection data are collected every 30 seconds or 60 seconds on average, and the concentration index of the dissolved gas in the solution and the corresponding PH value set by the user are reached, thereby ensuring an ideal growth environment for the cells.
Further, the process for adjusting the physical and chemical indexes in the production process comprises the following steps:
s1: firstly, aseptically sampling a reactor;
s2: secondly, transferring the sample to a monitoring station;
s3: checking basic indexes, wherein the indexes are respectively cell number, cell survival rate, glucose concentration, lactose concentration, amino acid substance concentration and other physical and chemical index detection;
s4: calculating the nutrient supplement amount and the production waste discharge amount;
s5: performing ATF waste liquid pumping;
s6: a new medium pump was performed.
Further, the steps S1 to S6 are carried out repeatedly, detection data are collected in sequence every 15 minutes or 30 minutes, effective nutrition levels verified by tests are achieved, sufficient nutrients for cell growth are guaranteed, and efficient use of culture media and the like is guaranteed.
Further, the culture product enrichment step is as follows:
s1: after production, firstly checking whether an enrichment process is started;
s2: detecting the concentration of a target product;
s3: detecting whether the residual solution in the reactor is larger than a set value;
s4: if the concentration value is less than the set value, the enrichment process is finished, and if the concentration value is greater than the set value, the next step is carried out;
s5: the ATF discharges the waste culture medium, and the process returns to step S3 again, and so on until the residual solution in the reactor is detected to be less than the set value.
Further, according to a product enrichment index set by a user, the enrichment operation of the product is carried out through the ATF module.
The invention has the beneficial effects that:
1. compared with the traditional reactor production mode, the method can basically avoid the problems of impact force and shearing force brought by a stirring paddle or carrier gas to cells in the production process, and can effectively avoid damage in the production process. The scale production of the animal suspension cells/human suspension cells which are not domesticated in the reactor becomes possible;
2. the special external gas and nutrient exchange unit can greatly improve the nutrient injection flux and the exchange efficiency. Compared with the traditional culture mode, the method can improve the cell culture density and the total yield by orders of magnitude.
Drawings
FIG. 1 is a schematic diagram of a closed loop reactor for exchanging biological products for external use according to the present invention;
FIG. 2 is a control diagram of the dissolved gas concentration of the culture solution in a closed loop external exchange bioproduct reactor of the present invention;
FIG. 3 is a diagram showing the control of the physicochemical indices of the culture solution in a closed-loop reactor for external exchange of biological products according to the present invention;
FIG. 4 is a diagram of the process of enrichment of culture products in a closed loop external exchange bioproduct reactor of the present invention;
FIG. 5 is a diagram of the reactor body of a closed loop external exchange bioproduct reactor of the present invention;
FIG. 6 is a schematic view of the upper main circulation incident structure of the reactor body of a closed circulation reactor for exchanging biological products;
FIG. 7 is a view showing the bottom main circulation discharge structure of the reactor body of a closed circulation external exchange bioproduct reactor according to the present invention;
FIG. 8 shows the efficient vortex mixing efficiency in the reactor body of a closed cycle external exchange bioproduct reactor of the present invention
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by a person of ordinary skill in the art without creative efforts based on the embodiments of the present invention belong to the protection scope of the present invention.
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Several embodiments of the invention are presented in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
Referring to fig. 1 to 8, the present invention provides a technical solution:
a closed circulation external exchange biological product reactor comprises a bioreactor body, a main circulation pipeline, a main circulation channel peristaltic pump, a liquid level sensor, a gas escape channel, a filter, a culture medium injection three-way pipeline, a gas exchange module, an O2/N2 channel, a dissolved oxygen and PH sensor group, a culture medium valve, a culture medium pump body, a culture medium storage unit, an ATF (automatic transfer function) matching module, an aseptic sample collection channel, automatic sample transfer and an online detection equipment group.
In order to further improve the use function of the closed-cycle external exchange biological product reactor, the corresponding functions are as follows:
the bioreactor body: the major sites for cell growth and the continuous maintenance of cells in suspension. The nutrient substances after gas exchange and culture medium injection are effectively diffused in the reactor;
a main circulation pipeline: the solution in the circulating main reactor is subjected to dissolved oxygen and carbon dioxide discharge exchange in a gas exchange module, and a new culture medium is injected into a culture solution tee joint in an exchange way;
main circulation channel peristaltic pump: used for driving the solution in the bioreactor to circulate among the main circulation pipeline, the gas exchanger and the culture medium adding channel;
a liquid level sensor: the device is used for continuously detecting the height of the liquid level in the main reactor and is used for closed-loop control of the liquid amount of used culture medium discharge and new culture medium addition;
gas escape channels and filters: during the production process, the pressure in the reactor rises due to the escape of dissolved gases and the volatilization of liquids. The channel discharges the excessive gas;
injecting a culture medium into the three-way pipeline: when the culture medium is added, the culture medium pumping pipeline and the main circulation pipeline are communicated, new culture medium enters the reactor main body, and when the tee joint is switched, the culture medium does not enter the main circulation pipeline;
a gas exchange module: the special membrane type gas exchange unit is used for carrying out efficient gas exchange. When the liquid passes through, O2 provided by a special pipeline is efficiently dissolved into the culture solution, and simultaneously, excessive dissolved CO2 in the culture solution is discharged through the introduction and dissolution of N2, and the pH value of the culture solution is adjusted through gas exchange;
O2/N2 channel: for supplying the sterilized gas subjected to the strict filtration treatment to the gas exchanger for culture solution dissolved gas exchange;
dissolved oxygen and PH sensor set: by measuring the dissolved oxygen concentration and the PH value before/after gas exchange, dynamically adjusting the liquid flow of the main circulation passing through the gas exchanger, the proportion of O2/N2 and the pressure of O2/N2, and realizing the dynamic adjustment of the dissolved oxygen concentration and the PH value based on target set values;
culture medium valve: connecting or closing the connection of the culture medium and the main circulation pipeline;
culture medium pump body: pumping the culture medium into the main circulation channel;
a medium storage unit: storing the culture medium for mass production for a long time in a storage environment of 4 ℃ degrees;
ATF enrichment unit: in the production process, continuously or periodically discharging the used culture medium, filtering the enriched cells, and returning the cells enriched in the module to the culture system again according to the requirement;
ATF matching module: used culture medium used for collecting the ATF is discharged to a waste liquid collecting barrel, and the ATF module is isolated from an external channel when the ATF module is not in work;
sterile sample collection channel: a culture solution sample in the reactor body is regularly and aseptically collected and is used for comprehensive index detection and culture liquid online adjustment, and the continuous control of main indexes in the production process is realized;
automatic sample transfer: after aseptically sampling to a sample tube, transferring the sample to each detection device on a test bench;
an online detection device group: the method is used for online production index detection and realizes closed-loop control of main biochemical indexes.
In order to further improve the use function of the closed circulation external exchange biological product reactor, the pH values in the dissolved oxygen and pH sensor groups are absolute values and difference values, and the liquid flow rate is the liquid rate; the dissolved oxygen and PH sensor group comprises a concentration detection unit before gas exchange and a concentration detection unit after gas exchange; the ATF matching module comprises an ATF valve, an ATF pump body, an ATF pipeline and an ATF waste liquid collecting unit; the sterile sample collecting channel comprises a sampling joint and a sampling valve; the online detection equipment group comprises a cell counting module and a biochemical index detection module.
In order to further improve the use function of the closed circulation external exchange biological product reactor, the dissolved gas concentration adjusting steps in the production process are as follows:
s1: first, DO measurement before gas exchange;
s2: secondly, measuring the PH before gas exchange;
s3: measuring DO after gas exchange;
s4: measuring the PH after gas exchange;
s5: obtaining the difference value of DO and PH before and after gas exchange;
s6: calculating O2、N2Exchange efficiency target of (2);
s7: calculating the flow rate of the culture liquid in the gas exchanger;
s8: calculating O2、N2Flow rate in the pipeline;
s9: controlling and adjusting the flow of the culture liquid; control adjustment O2/N2And (4) flow rate.
In order to further improve the use function of the closed-cycle external exchange biological product reactor, the steps S1 to S9 are repeated, detection data are collected every 30 seconds or 60 seconds on average, the concentration index of dissolved gas in the solution set by a user and the corresponding pH value are reached, and an ideal growth environment of cells is guaranteed.
In order to further improve the use function of the closed circulation external exchange biological product reactor, the physicochemical index adjustment process of the production process is as follows:
s1: firstly, aseptically sampling a reactor;
s2: secondly, transferring the sample to a monitoring station;
s3: checking basic indexes, wherein the indexes are respectively cell number, cell survival rate, glucose concentration, lactose concentration, amino acid substance concentration and other physical and chemical index detection;
s4: calculating the nutrient supplement amount and the production waste discharge amount;
s5: performing ATF waste liquid pumping;
s6: a new medium pump was performed.
In order to further improve the use function of the closed-cycle external exchange biological product reactor, the steps S1 to S6 are carried out repeatedly, the detection data are collected in sequence every 15 minutes or 30 minutes, the effective nutrition level verified by the test is achieved, sufficient nutrients for cell growth are guaranteed, and meanwhile, efficient use of culture media and the like is guaranteed.
In order to further improve the use function of the closed circulation external exchange biological product reactor, the culture product enrichment step is as follows:
s1: after production, firstly checking whether an enrichment process is started;
s2: detecting the concentration of a target product;
s3: detecting whether the residual solution in the reactor is larger than a set value;
s4: if the concentration value is less than the set value, the enrichment process is finished, and if the concentration value is greater than the set value, the next step is carried out;
s5: the ATF discharges the waste culture medium, and the process returns to step S3 again, and so on until the residual solution in the reactor is detected to be less than the set value.
In order to further improve the use function of the closed-cycle external exchange biological product reactor, the product is enriched through the ATF module according to the product enrichment index set by a user. The main circulation of the main reactor adopts the structural design of oblique angle incidence and oblique angle discharge, and stable circulating vortex is formed in the reactor to form effective system circulation.
In the reactor, the high liquid flow rate is maintained in all areas by the vortex fluid motion, and the cells and the production products are prevented from being adhered and accumulated on the inner wall of the reactor. The cells and the production products are ensured to be uniformly distributed in the reactor;
the cells and the production products are continuously kept in a suspension state through vortex fluid, effective diffusion exchange of nutrients is kept, and the excellent growth state of the cells is kept;
the device can ensure the effective circulation of liquid and avoid the huge impact force and shearing force to cells when the stirring paddle in the traditional reactor works. The maximum shearing force generated by the reactor is less than 0.1Pa, the shearing force tolerance of the sensitive animal cells produced by the target is greater than 0.4Pa, the damage of the shearing force to the cells in the production process can be effectively avoided, and the excellent growth state of the cells is ensured;
a gas leading-in device in the traditional reactor is cancelled, so that one of the potential pollution generation reasons caused by bubbles in the production process is greatly reduced, and the pollution generation probability is reduced. Meanwhile, the influence of the impact force generated by the burst of the bubbles in the fluid on the cells is avoided, and an ideal environment is provided for the growth of the cells;
according to the invention, the water content and the net content of the muck are firstly measured, so that the overloading of the muck truck caused by the over-standard water content of the muck truck can be effectively prevented when the muck truck carries the muck, the traffic accident is caused, the running safety of the muck truck is ensured, the problem that the muck truck is weighed after loading in the prior art is also replaced, and the excessive muck needs to be discharged if the muck truck is overweight, which is very troublesome.
The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (9)
1. A closed-cycle external-exchange biological product reactor is characterized by comprising a bioreactor body, a main circulation pipeline, a main circulation channel peristaltic pump, a liquid level sensor, a gas escape channel and filter, a culture medium injection three-way pipeline, a gas exchange module, an O2/N2 channel, a dissolved oxygen and PH sensor group, a culture medium valve, a culture medium pump body, a culture medium storage unit, an ATF (automatic transfer function) matching module, a sterile sample collection channel, automatic sample transfer and an online detection equipment group.
2. A closed cycle external exchange bioproduct reactor according to claim 1, characterized by the corresponding functions as follows:
the bioreactor body: the major sites for cell growth and the continuous maintenance of cells in suspension. The nutrient substances after gas exchange and culture medium injection are effectively diffused in the reactor;
a main circulation pipeline: the solution in the circulating main reactor is subjected to dissolved oxygen and carbon dioxide discharge exchange in a gas exchange module, and a new culture medium is injected into a culture solution tee joint in an exchange way;
main circulation channel peristaltic pump: used for driving the solution in the bioreactor to circulate among the main circulation pipeline, the gas exchanger and the culture medium adding channel;
a liquid level sensor: the device is used for continuously detecting the height of the liquid level in the main reactor and is used for closed-loop control of the liquid amount of used culture medium discharge and new culture medium addition;
gas escape channels and filters: during the production process, the pressure in the reactor rises due to the escape of dissolved gases and the volatilization of liquids. The channel discharges the excessive gas;
injecting a culture medium into the three-way pipeline: when the culture medium is added, the culture medium pumping pipeline and the main circulation pipeline are communicated, new culture medium enters the reactor main body, and when the tee joint is switched, the culture medium does not enter the main circulation pipeline;
a gas exchange module: the special membrane type gas exchange unit is used for carrying out efficient gas exchange. When the liquid passes through, O2 provided by a special pipeline is efficiently dissolved into the culture solution, and simultaneously, excessive dissolved CO2 in the culture solution is discharged through the introduction and dissolution of N2, and the pH value of the culture solution is adjusted through gas exchange;
O2/N2 channel: for supplying the sterilized gas subjected to the strict filtration treatment to the gas exchanger for culture solution dissolved gas exchange;
dissolved oxygen and PH sensor set: by measuring the dissolved oxygen concentration and the PH value before/after gas exchange, dynamically adjusting the liquid flow of the main circulation passing through the gas exchanger, the proportion of O2/N2 and the pressure of O2/N2, and realizing the dynamic adjustment of the dissolved oxygen concentration and the PH value based on target set values;
culture medium valve: connecting or closing the connection of the culture medium and the main circulation pipeline;
culture medium pump body: pumping the culture medium into the main circulation channel;
a medium storage unit: storing the culture medium for mass production for a long time in a storage environment of 4 ℃ degrees;
ATF enrichment unit: in the production process, continuously or periodically discharging the used culture medium, filtering the enriched cells, and returning the cells enriched in the module to the culture system again according to the requirement;
ATF matching module: used culture medium used for collecting the ATF is discharged to a waste liquid collecting barrel, and the ATF module is isolated from an external channel when the ATF module is not in work;
sterile sample collection channel: a culture solution sample in the reactor body is regularly and aseptically collected and is used for comprehensive index detection and culture liquid online adjustment, and the continuous control of main indexes in the production process is realized;
automatic sample transfer: after aseptically sampling to a sample tube, transferring the sample to each detection device on a test bench;
an online detection device group: the method is used for online production index detection and realizes closed-loop control of main biochemical indexes.
3. The closed loop, external exchange bioproduct reactor of claim 1, wherein: the pH values of the dissolved oxygen and the PH sensor group are absolute values and difference values, and the liquid flow rate is the liquid rate; the dissolved oxygen and PH sensor group comprises a concentration detection unit before gas exchange and a concentration detection unit after gas exchange; the ATF matching module comprises an ATF valve, an ATF pump body, an ATF pipeline and an ATF waste liquid collecting unit; the sterile sample collecting channel comprises a sampling joint and a sampling valve; the online detection equipment group comprises a cell counting module and a biochemical index detection module.
4. The closed loop, external exchange bioproduct reactor of claim 1, wherein the process dissolved gas concentration adjustment step is as follows:
s1: first, DO measurement before gas exchange;
s2: secondly, measuring the PH before gas exchange;
s3: measuring DO after gas exchange;
s4: measuring the PH after gas exchange;
s5: obtaining the difference value of DO and PH before and after gas exchange;
s6: calculating O2、N2Exchange efficiency target of (2);
s7: calculating the flow rate of the culture liquid in the gas exchanger;
s8: calculating O2、N2Flow rate in the pipeline;
s9: controlling and adjusting the flow of the culture liquid; control adjustment O2/N2And (4) flow rate.
5. The closed loop, external exchange bioproduct reactor of claim 4, wherein: wherein, the steps S1 to S9 are repeated, the detection data are collected once every 30 seconds or 60 seconds on average, and the concentration index of the dissolved gas in the solution set by a user and the corresponding PH value are reached, thereby ensuring that the cells have ideal growth environment.
6. The closed loop external exchange bioproduct reactor of claim 1, wherein the process physicochemical index adjustment process is as follows:
s1: firstly, aseptically sampling a reactor;
s2: secondly, transferring the sample to a monitoring station;
s3: checking basic indexes, wherein the indexes are respectively cell number, cell survival rate, glucose concentration, lactose concentration, amino acid substance concentration and other physical and chemical index detection;
s4: calculating the nutrient supplement amount and the production waste discharge amount;
s5: performing ATF waste liquid pumping;
s6: a new medium pump was performed.
7. The closed loop, external exchange bioproduct reactor of claim 6, wherein: the steps S1 to S6 are carried out repeatedly, detection data are collected in sequence every 15 minutes or 30 minutes, the effective nutrition level verified by tests is achieved, sufficient nutrients for cell growth are guaranteed, and efficient use of culture media and the like is guaranteed.
8. The closed loop, external exchange bioproduct reactor of claim 1, wherein the culture product enrichment step is as follows:
s1: after production, firstly checking whether an enrichment process is started;
s2: detecting the concentration of a target product;
s3: detecting whether the residual solution in the reactor is larger than a set value;
s4: if the concentration value is less than the set value, the enrichment process is finished, and if the concentration value is greater than the set value, the next step is carried out;
s5: the ATF discharges the waste culture medium, and the process returns to step S3 again, and so on until the residual solution in the reactor is detected to be less than the set value.
9. The closed loop, external exchange bioproduct reactor of claim 8, wherein: wherein, the enrichment operation of the product is carried out through the ATF module according to the product enrichment index set by the user.
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