A rolled fixed-bed bioreactor suitable for vaccine and antibody production

Abstract

The present invention discloses a rolled fixed-bed bioreactor suitable for the production of vaccines and antibodies. A fiber roll rolled from a thermoplastic resin is arranged in a fiber cavity of the fixed-bed reactor. A skeleton support is arranged on the outer transverse end surface of the fiber roll. The lower part of the fiber cavity is connected to an impeller cavity, and an impeller is arranged in the impeller cavity. The present invention transforms the traditional stacked sheet fiber carriers into a rolled fiber carrier fixed bed. The fiber roll makes the direction of the fixed bed matrix the same as the flow direction of the liquid, the liquid circulation during cell inoculation is better, the inoculation uniformity is better, the wall adhesion homogeneity is better during the culture process, the cells have good uniformity in nutrient and oxygen consumption, and the cultured cells can be conveniently sampled by arranging easy-to-tear long fiber strips to observe the growth status of the cells. At the same time, the sampling device is specially designed to greatly reduce the risk of bacterial contamination caused by sampling.

Description

Technical Field

[0001] The present invention relates to the technical field of animal cell culture, in particular to a roll-type fixed-bed bioreactor suitable for vaccine and antibody production. Background Art

[0002] Fixed-bed bioreactors offer a simpler structure, better culture results, and ease of scale-up. Compared to traditional stirred-bed bioreactors, they significantly simplify the adherent cell culture process and are widely used in gene therapy, vaccine, and virus production. However, existing fixed-bed bioreactors cannot effectively monitor cell status (such as cell number and cell attachment uniformity) during the culture process. Summary of the Invention

[0003] The technical problem to be solved by the present invention is to overcome the defect in the existing fixed bed reactor that the cell attachment uniformity and other cell states cannot be effectively observed, and to provide a roll-type fixed bed bioreactor suitable for vaccine and antibody production.

[0004] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0005] A rolled fixed-bed bioreactor suitable for vaccine and antibody production, comprising a fixed-bed reactor, a liquid storage system, and a detection system;

[0006] The fixed bed reactor includes a fiber cavity and a multi-layer fiber roll inside it. A skeleton support is provided on the outer transverse end surface of the fiber roll. The lower part of the fiber cavity is connected to the impeller cavity. The impeller cavity is provided with an impeller. The rotation of the impeller can drive the culture medium in the fiber cavity to circulate from bottom to top, so that the culture medium in the fiber cavity is evenly distributed and the culture effect is better. The upper part of the fiber cavity is connected to the fiber cavity cover; the fiber cavity cover is provided with several sampling port covers and inoculation port covers connected to the fiber cavity; the material of the fiber roll is PET or PE of food and medical grade, and the material of the fiber cavity is PP or PE of food and medical grade.

[0007] The liquid storage system includes a fresh culture medium bottle and a perfusion bottle; the fresh culture medium bottle and the perfusion bottle are connected to the fixed bed reactor through a silicone tube and a peristaltic pump;

[0008] The outermost fiber roll is provided with a plurality of easy-tear portions, which divide the outermost fiber roll into a plurality of fiber strips. Preferably, the easy-tear portion can be a row of pre-punched holes to form long fiber strips that are easy to tear off. The long fiber strips are connected to the sampling device, which can conveniently sample the cells attached to the carrier during the culture process.

[0009] Magnets are installed inside the sampling port cover and on the sampling rod. The sampling port cover is connected to the sampling rod through magnet adsorption, and the end of the sampling rod is connected to the fiber strip. When sampling, separate the sampling device and the cover, and tighten the sampling cover to complete the sampling conveniently.

[0010] Furthermore, an inverted conical filter is installed above the fiber cavity. The impeller is magnetically driven by a motor at the bottom of the fixed-bed reactor. The culture medium flows from the bottom of the fixed-bed reactor to the top, passing through the filter, forming a thin film that flows down the outer wall of the fiber cavity. This provides a large mass transfer area and enables efficient gas exchange.

[0011] Furthermore, the sampling rod is made of PP, the fiber strip is made of PET, and the sampling rod and the fiber strip are connected by thermal bonding.

[0012] Furthermore, the system also includes a detection and control system, which is used to detect and adjust the pH, dissolved oxygen value and temperature in the fixed bed reactor so that the culture medium therein is at a set pH, dissolved oxygen and temperature.

[0013] Further, the detection and control system includes a pH control module, a temperature control module and a dissolved oxygen control module, wherein the pH control module is composed of a pH electrode and a pH actuator module, and the temperature control module is composed of a temperature electrode and a temperature execution module. The pH actuator module is composed of a carbon dioxide control module and a sodium bicarbonate control module. The pH electrode is electrically connected to the pH electrode interface. When the pH electrode detects that the pH is higher than the set value, the carbon dioxide control module increases the carbon dioxide ventilation volume. When the pH is lower than the set value, the sodium bicarbonate control module connected to sodium bicarbonate starts (peristaltic pump peristaltic start), adds an appropriate amount of alkali solution, and when the temperature electrode detects that the temperature is lower than the set temperature, the temperature execution module starts and heats the alkali solution bottle to the set temperature. Preferably, the temperature execution module is a heating blanket provided on the outside of the alkali solution bottle, and the temperature of the liquid in the alkali solution bottle can be regulated by starting or closing the heating blanket. The dissolved oxygen control module is composed of a dissolved oxygen electrode and a dissolved oxygen actuator module, and the dissolved oxygen actuator module realizes DO control by controlling the flow of air / oxygen.

[0014] The fixed-bed reactor features multiple openings, three of which are equipped with pH, ​​dissolved oxygen, and temperature electrodes. These electrodes are coupled to air / oxygen and carbon dioxide flow control units, enabling pH and DO control by adjusting gas flow rates. The other two openings are the air inlet and exhaust ports. The inlet is connected to a pressure sensor to monitor system blockage, while the exhaust is connected to a filter equipped with a heating plate to prevent moisture from wetting the filter during prolonged incubation. Air, oxygen, or carbon dioxide enters through the inlet, where a pressure sensor connected to the control system is located.

[0015] Furthermore, when the pressure sensor detects an abnormal pressure in the fixed bed reactor, the control system cuts off ventilation and issues an alarm signal by disconnecting the solenoid valve connected to the dissolved oxygen actuator module or the carbon dioxide control module.

[0016] Beneficial effects: The present invention improves the fixed bed bioreactor by changing the traditional fixed bed of stacked sheet fiber carriers into a rolled fiber carrier fixed bed. The fiber roll makes the direction of the fixed bed matrix the same as the direction of the liquid flow, and the liquid circulation during cell inoculation is better. Compared with the stacked fixed bed, there is no obstruction, so the rolled fixed bed has better uniformity during cell inoculation, and the wall adhesion homogeneity during the culture process is better. In addition, the traditional stacked rolled fixed bed has a large resistance to mass transfer and oxygen transfer between bed layers. When the culture medium circulates from top to bottom, when the culture medium flows to the upper end of the fixed bed, this part of nutrients, oxygen, etc. will be relatively reduced. Moreover, each layer of the rolled fixed bed is attached, and the uniformity of cell consumption of nutrients and oxygen is better than that of the stacked type. The utilization of nutrients by cells in each layer is relatively balanced.

[0017] In addition, the present invention provides an easy-to-tear long fiber strip on the outside of the fiber roll. The long fiber strip is connected to the sampling device, which can conveniently sample the cultured cells and observe the growth status of the cells. At the same time, the sampling device is specially designed to greatly reduce the risk of contamination caused by sampling. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0019] Figure 1 Schematic diagram of the three-dimensional structure of the bioreactor of the present invention;

[0020] Figure 2 It is a schematic diagram of the explosion structure of the fixed-bed bioreactor of the present invention;

[0021] Figure 3 It is a schematic structural diagram of a sampling device of a fixed-bed bioreactor of the present invention;

[0022] Figure 4 It is a partial structural schematic diagram of the bioreactor of the present invention;

[0023] Figure 5 It is a schematic diagram of the principle of a fixed-bed bioreactor of the present invention;

[0024] Figure 6 yes Figure 1 Enlarged view of point A in the middle;

[0025] Figure 7 It is a module diagram of the detection and control system. DETAILED DESCRIPTION

[0026] The preferred embodiments of the present invention are described below with reference to the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0027] Example

[0028] A rolled fixed-bed bioreactor suitable for vaccine and antibody production comprises a fixed-bed reactor 1, a liquid storage system and a detection and control system. The detection and control system is connected to a computer via a data line 13 to control the entire system.

[0029] The fixed bed reactor 1 includes a sampling rod 5, a fiber roll 14, a fiber strip 6, a fiber cavity 7, an impeller 8, a sampling port cover 10, and an inoculation port cover 11. A skeleton support 15 is provided on the outer transverse end surface of the fiber roll. The upper cylindrical portion 51 of the sampling rod 5 and the interior of the sampling port cover 11 are both embedded with magnets, which can be adsorbed and connected to each other. The lower end of the sampling rod 5 is connected to the fiber strip 6, and the fiber strip 6 can be torn off from the fiber roll 5 through the easy-to-tear portion. When sampling, after opening the sampling port cover 10, use tweezers to separate the sampling rod 5 and the sampling port cover 10, and then tear off the fiber strip 6 to complete the sampling. The exposure time and exposure area during the sampling process are short, and no external sampling tools need to be inserted, which greatly reduces the probability of contamination. The sampling process is simple and easy to operate, and does not affect the progress of the culture process. The fiber roll is made of food and medical grade PET, and the fiber cavity is made of food and medical grade PP or PE.

[0030] An inverted conical filter screen 16 is installed above the fiber cavity 7 through a support 17. The impeller 8 is magnetically driven by a motor at the bottom of the fixed-bed reactor 1. The culture medium flows from the bottom to the top of the fixed reactor 1, passes through the filter screen 16, and forms a thin film that flows down the outer wall of the fiber cavity 7. This provides a large mass transfer area and enables efficient gas exchange.

[0031] Figure 6 yes Figure 1In the enlarged view at point A, a pressure sensor 12 is connected to the pipe at the fixed-bed reactor's air inlet. When the pressure sensor 12 detects a pressure rise reaching a certain value, the detection and control system shuts off ventilation and issues an alarm. The fixed-bed reactor also has an exhaust port equipped with a filter mounted on a heater plate 18. This ensures that water vapor from the exhaust port is discharged as steam and does not condense into water and wet the filter, thereby preventing bacterial contamination. The dissolved oxygen electrode interface 19, temperature electrode interface 20, heater plate interface 21, pH electrode interface 22, and pressure sensor interface 23 are all connected to the detection and control system, providing electrical connection and control for the dissolved oxygen electrode, temperature electrode, heater plate, pH electrode, and pressure sensor, respectively. If the pressure sensor 12 detects an abnormal internal pressure, the detection and control system shuts off ventilation and issues an alarm by disconnecting the solenoid valve connected to the dissolved oxygen actuator module or carbon dioxide control module. Luer Ordering info Press-S-000 pressure sensors are recommended.

[0032] The liquid storage system includes a fresh culture medium bottle 2, an alkaline solution bottle 3, and a perfusion bottle 4. These bottles are connected to the fixed-bed reactor 1 via silicone tubing and a peristaltic pump 9. During operation, the peristaltic pump 9 periodically extracts a portion of the culture medium from the fixed-bed reactor 1 and adds it to the perfusion bottle 4. It then extracts an equal amount of fresh culture medium from the fresh culture medium bottle 2 and adds it to the fixed-bed reactor 1, maintaining a relatively constant liquid volume in the fixed-bed reactor. The detection and control system uses feedback from the pH and dissolved oxygen electrodes to control the aeration volume and the flow rate of the alkaline solution, thereby controlling the pH and DO values ​​of the culture medium.

[0033] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art will be able to modify the technical solutions described in the aforementioned embodiments or substitute equivalents for some of the technical features. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A scroll-type fixed-bed bioreactor suitable for vaccine and antibody production, comprising a fixed-bed reactor and a liquid storage system, characterized in that: The fixed bed reactor includes a fiber cavity, a fiber roll made of thermoplastic resin is arranged in the fiber cavity, a skeleton support is arranged on the outer transverse end surface of the fiber roll, the lower part of the fiber cavity is connected to the impeller cavity, and the impeller cavity is provided with an impeller; the upper part of the fiber cavity is connected to the fiber cavity cover; The liquid storage system includes a fresh culture medium bottle and a perfusion bottle; the fresh culture medium bottle and the perfusion bottle are connected to the fixed bed reactor through a silicone tube and a peristaltic pump; The fiber cavity cover is provided with a plurality of sampling port covers and inoculation port covers connected to the fiber cavity, and a sampling rod is provided in the fiber cavity at the sampling port cover; a plurality of easy-tear parts are provided on the outermost fiber roll, and the easy-tear parts divide the outermost fiber roll into a plurality of fiber strips; An inverted cone-shaped filter screen is provided above the fiber cavity; The inside of the sampling port cover and the sampling rod are both equipped with magnets. The sampling port cover is connected to the sampling rod through magnet adsorption, and the end of the sampling rod is connected to the fiber strip. The sampling rod is made of PP, the fiber strip is made of PET, and the sampling rod and the fiber strip are connected by thermal bonding.

2. The wound fixed-bed bioreactor suitable for vaccine and antibody production according to any one of claim 1, characterized in that: The material of the fiber roll is food and medical grade PET or PE, and the material of the fiber cavity is food and medical grade PP or PE.

3. The scroll-type fixed-bed bioreactor suitable for vaccine and antibody production according to claim 1, characterized in that: The system also includes a detection and control system, which is used to detect and adjust the pH, dissolved oxygen value and temperature in the fixed bed reactor so that the culture medium in the fixed bed reactor is at the set pH, dissolved oxygen and temperature.

4. The wound fixed-bed bioreactor suitable for vaccine and antibody production according to claim 3, characterized in that: The detection and control system includes a pH control module, a temperature control module and a dissolved oxygen control module. The pH control module is composed of a pH electrode and a pH actuator module. The temperature control module is composed of a temperature electrode and a temperature actuator module. The pH actuator module is composed of a carbon dioxide control module and a sodium bicarbonate control module. The pH electrode is installed at the pH electrode interface. When the pH detection probe of the pH electrode detects that the pH is higher than the set value, the carbon dioxide control module increases the carbon dioxide ventilation volume. When the pH is lower than the set value, the sodium bicarbonate control module connected to sodium bicarbonate is started and an appropriate amount of alkali solution is added. When the temperature electrode detects that the temperature is lower than the set temperature, the temperature execution module is started to heat the alkali solution bottle to the set temperature. The dissolved oxygen control module is composed of a dissolved oxygen electrode and a dissolved oxygen actuator module. The dissolved oxygen actuator module realizes DO control by controlling the flow of air / oxygen.

5. The scroll-type fixed-bed bioreactor suitable for vaccine and antibody production according to claim 4, characterized in that: Air, oxygen or carbon dioxide enters through an air inlet on the fixed bed reactor, and a pressure sensor connected to a detection and control system is provided on the pipe at the air inlet; the fixed bed reactor is also provided with an exhaust port, a filter is provided at the exhaust port, and a heating plate is installed on the filter.

6. The scroll-type fixed-bed bioreactor suitable for vaccine and antibody production according to claim 5, characterized in that: When the pressure sensor detects abnormal pressure in the fixed bed reactor, the detection and control system cuts off ventilation and sends an alarm signal by disconnecting the solenoid valve connected to the dissolved oxygen actuator module or the carbon dioxide control module.

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