RU2797021C1 - Asymmetric conical bioreactor system and method for its use - Google Patents

Abstract

FIELD: biotechnology; medicine.

SUBSTANCE: asymmetric conical bioreactor includes a hermetically sealed rigid vessel containing a top and a bottom lid, the top lid being connected to the vessel by means of a flange, the inner surface of the vessel being made in the form of a truncated cone, having a rectangular trapezoid in cross section, and on the outer surface of the vessel at least at least two mounting holes for installing sensors for monitoring the process, on the surfaces of the vessel, covers and flange there are seats for installing one gas inlet port, gas outlet, sampling port, culture introduction port, reagents injection port, culture collection port and waste disposal port. In this variant, the bioreactor also includes at least one membrane inside the vessel, which divides the internal volume of the vessel into two spaces, one of which is an area of gas accumulation, where a drain valve with an extension to remove the air bubble is located. Also described is a bioreactor system for cell culture, including the specified bioreactor.

EFFECT: group of inventions makes it possible to increase the viability of cultured cells while improving mass transfer characteristics.

31 cl, 12 dwg, 2 ex

Description

The field of technology to which the invention belongs

The invention relates to biotechnology and medicine, in particular for cell cultivation. The asymmetric conical bioreactor is used for the implementation of biotechnological processes occurring in the liquid phase during the cultivation of cells, microorganisms and viruses, the asymmetric conical bioreactor system is designed for the cultivation of cells, microorganisms and viruses. The proposed system can be presented both as an independent device and as part of a biotechnological complex, and can be used for growing cell cultures that are sensitive to mechanical stress, growing cells for traditional fermenters, for scientific and industrial research, for high-quality production of cells for industrial bioreactors, for the production of biomedical products.

BACKGROUND OF THE INVENTION Bioreactors can be used to produce products such as pharmaceuticals, vaccines, antibodies, and the like by culturing cells, organisms, cultures, and the like under a controlled environment. Increasingly, disposable bioreactors are being used that comprise a support structure and a disposable container configured to fit within the support structure. The use of systems with disposable containers avoids the risks of infection and contamination between processes.

There are a number of design solutions for disposable bioreactors. Applikon Biotechnology (Netherlands) and Stedim Inc. (France) offer the Appliflex® Disposable Bioreactor System, which uses 50-liter flexible culture bags that are designed to be placed on a motorized platform that rock the bag to provide aeration and agitation of the culture medium. A similar disposable bioreactor is offered by Wave Biotech, LLD (Somerset, NJ, USA), which uses culture bags up to 1000 L, which are also aerated and agitated using a motorized platform. Aeration and agitation is provided by a reusable paddle impeller attached to a complex drive device inserted into the culture bag.

RF Patent No. 2468076 discloses a disposable device for cultivating and collecting plant tissue and/or cells, which contains a non-rigid container with a volume of at least 400 liters and is equipped with gas exchange channels and a collection channel; a support structure for supporting said device, wherein said support structure includes a rigid bottom having the shape of a truncated cone. This configuration of the specially designed work support affects the overall viability of the cultures by avoiding the formation of cell pellets that can enhance cell decay and death. Mixing is carried out by supplying air or gas for aeration and mixing.

In all of the aforementioned systems, the support and aeration/mixing systems are complex, costly, highly targeted, and limited in power. So, although the reactor vessel (container) itself can be disposable, the use of these systems requires expensive adjustment and maintenance.

The closest analogue of the claimed invention is US patent No. 6544788. Cells are grown in a plastic bag that is rocked and ventilated on a mechanical platform. The bioreactor bag contains a filter that allows liquid to be removed from the bioreactor without cell loss. Nutrients can be added through another port. The perfusion filter is designed in such a way that it can move freely on the surface of the liquid. The filter moves rapidly across the surface as a result of the rocking of the bioreactor, and this rocking movement of the filter prevents it from clogging. The control system ensures continuous operation for several weeks.

In addition to insufficient productivity, the economics of using this kind of reactor in many cases cannot be ensured without sufficient scalability. An increase in scale in this situation is possible only at the cost of increasing complexity and reducing the cost-effectiveness of use, through the additional use of complex technical solutions. This system does not allow starting processes from small volumes (which saves expensive reagents) and gradually increasing the volume as cells expand, as well as operating the same system in manual, semi-automatic and fully automatic modes.

Thus, there is a constant need to improve existing systems and devices for the culture of cells, microorganisms and viruses in order to be able to build the necessary processes in manual, semi-automatic and fully automatic modes, as well as to scale systems and devices without changing the configuration of the entire bioreactor system with an increase in volume. as cells expand, and improve cost efficiency.

The present application is devoted to the solution of this problem.

The essence of the invention

The technical problem solved by the present invention is

- creation of a bioreactor for the implementation of biotechnological processes occurring in the liquid phase during the cultivation of cells, microorganisms and viruses, which could be used as an independent device, as well as as part of an automated biotechnological complex, and which would have the ability to reduce the number of reagents used and increase the viability of cellular cultures;

- creation of a flexible bioreactor system that will allow building the necessary process in manual, semi-automatic and fully automatic modes;

- the ability to start processes from small volumes, which allows saving expensive reagents and gradually increasing the volume as the cells expand;

- supply gases to the bioreactor with the required parameters.

At the same time, the developed design of the bioreactor and the bioreactor system that uses it must be easy to handle, must meet the high technical requirements of the pharmaceutical industry in terms of cleaning and sterility.

The technical result achieved by using the invention is to increase the viability of a cultivated cell culture with its efficient reproduction and at the same time improve mass transfer characteristics.

The technical result is achieved due to the fact that, according to the first aspect of the invention, the bioreactor for carrying out biotechnological processes occurring in the liquid phase during the cultivation of cells, microorganisms and viruses, includes a sealed rigid vessel for cultivation, while the inner surface of the vessel is made in the form of a truncated cone and has a rectangular trapezoid in cross section with a smaller base downwards of the vessel, and on the outer surface of the vessel there are at least two mounting holes for installing sensors for monitoring the biotechnological process so that when installing the sensors, they are completely immersed in the cultivated medium when filling the vessel with it.

Also, the technical result is achieved due to the fact that, according to the second aspect of the invention, the bioreactor for carrying out biotechnological processes occurring in the liquid phase during the cultivation of cells, microorganisms and viruses, includes a sealed rigid vessel for cultivation, while the inner surface of the vessel is made in the form of a truncated cone and has in cross section, a rectangular trapezoid with a smaller base downwards of the vessel, and at least two landing holes are placed on the outer surface of the vessel for installing sensors for monitoring the biotechnological process so that when installing the sensors, they are completely immersed in the cultivated medium when filling the vessel with it, at the same time, at least one membrane is installed inside the vessel, which divides the internal volume of the vessel into at least two spaces, one of which is the area of the gas accumulation space, and in the area of the gas accumulation space there is a drain valve with an extension to remove the air bubble from this space .

In some embodiments of the invention, the vessel has a top cover and a bottom cover, while the internal space of the vessel, bounded by the internal surface of the vessel, is divided into three chambers by membranes, which form the upper space, bounded by the internal surface of the top cover and the first membrane, the lower space, bounded by the internal the bottom cover surface and the second membrane, the intermembrane space bounded by the first and second membranes.

In some embodiments of the invention, the outer surface of the vessel has a predominantly vertical cylindrical shape with a beveled side wall, the vessel is sealed with lids on both ends.

In some embodiments of the invention, the vessel and covers have landing holes for installing ports, the geometry and functionality of which are selected in accordance with the requirements of the biotechnological process.

In some embodiments, biotech process monitoring sensors are disposable or reusable. At the same time, sensors for monitoring the biotechnological process are selected from: a dissolved oxygen sensor DO, an acid-base balance sensor PH.

In some embodiments of the invention, the bioreactor further includes disposable sensors placed in the vessel bores and selected from: a glucose sensor, a lactate sensor, a temperature sensor, a liquid level sensor, a pressure sensor, a biomass sensor.

In some embodiments of the invention, sensors for monitoring the biotechnological process are pre-installed.

In some embodiments of the invention, the internal volume of the vessel is from about 15 ml to 50,000 ml.

In some embodiments of the invention, each port corresponds to a port, and the ports are selected from the group: gas inlet port, gas outlet port, sampling port, culture injection port, reagent injection port, culture collection port, withdrawal port waste, a port for removing the air bubble from under the membrane.

In some embodiments of the invention, the material of the vessel and/or lids is a biocompatible material or a combination of biocompatible materials, wherein the biocompatible material is selected from the group: glass, polyethylene, stainless steel, polycarbonate, polysulfone.

Also, the technical result is achieved due to the fact that, according to the third aspect of the invention, the bioreactor system for cultivating cells, microorganisms and viruses includes the above-described bioreactor with a heating means, and a control tower equipped with a pendulum support mechanism capable of holding, rotating and rocking a vertically installed it contains a bioreactor vessel relative to its vertical axis during the biotechnological process, an electrical connection panel, a power supply unit, a control unit configured to execute commands from peripheral computing devices and / or a control panel integrated with a pendulum mechanism to set the current parameters of the biotechnological process, receive data on process trends from disposable or reusable sensors to obtain measurements of the physical, chemical or biological properties of the contents of the vessel during the biotechnological process, adjust the parameters of the biotechnological process depending on the measurements obtained by applying control signals to the mechanisms for supplying gas, nutrient media to the bioreactor vessel, configured to deliver gas, nutrient media to the bioreactor vessel during events in response to a control signal, to a heating means configured to change and maintain the temperature in the bioreactor vessel in response to a control signal, and to a pendulum mechanism configured to change the angle of inclination in response to a control signal.

In some embodiments, the implementation of the pendulum mechanism is made in the form of a clamping collar, mounted on the shaft of a stepper motor, mounted on a rack mounted on a base plate.

In some embodiments, the current parameters of the biotechnological process are selected from: the maximum speed and maximum angle of inclination of the bioreactor vessel from the vertical axis of the pendulum mechanism, the volume and temperature of the supplied reagents and their supply rate, the heating temperature of the contents of the vessel, the pressure in the vessel, pH, DO, Glu, Lac, Biomass.

In some embodiments, the control tower is equipped with peristaltic pumps to supply reactants to the bioreactor vessel.

In some embodiments, the implementation of the control panel is located on the body of the control tower and is made in the form of a touch screen connected to the control unit.

In some embodiments, the implementation of the control panel is configured to transmit data to the computing module, which issues commands to the control unit.

In some embodiments, peripheral computing devices are selected from: a laptop, a personal computer, a mobile phone, and are configured to run a web application on them to set the current parameters of the biotechnological process.

In some embodiments, the peripheral computing devices are configured to communicate data to a computing module that issues commands to a control unit.

In some embodiments, the electrical connection panel is configured to connect the pendulum mechanism, reusable or disposable sensors, valves, mechanisms for supplying reagents to the bioreactor vessel, and equipment necessary to ensure the biotechnological process.

In some embodiments, the power supply unit is configured to power the pendulum mechanism, peristaltic pumps, sensors, display, light and sound indication, fans, control device, pinch valves, fluid flow heater, heating means.

In some embodiments, LED indicators are located on the control tower body, associated with the control unit and signaling at least one of the events: an emergency; completion of the process; deviation of process indicators from the required ones; readiness of the equipment to start the process; finding the system in operation.

In some embodiments, the heating means is a heating jacket that is externally applied to the outer surface of the vessel to maintain the culture temperature of the bioreactor.

In some embodiments, the material of the bioreactor vessel is stainless steel, the vessel has at least one opening in the vessel wall for receiving a heating cartridge, and the heating means is a heating cartridge for heating the walls of the vessel inserted into one of these openings.

In some embodiments, the heating means is a heating cartridge inserted directly into the interior of the vessel.

In some embodiments, the heating means is an inline heater in the body of the bioreactor vessel.

In some embodiments, the heating means is a thermally conductive sprayed layer by vacuum deposition technology on the inner or outer surface of the vessel with wires soldered to the sprayed layer and brought out to the outside.

In some embodiments, the implementation of the system further includes a system for heating the fluid supplied to the reactor to the required temperature.

In some embodiments, a mounting plate is installed on the control tower system so that it is located in the area of the bioreactor vessel, to place on it the equipment required to ensure the process, selected from: pipeline, pinch valves, fittings, oxygenator, bubble traps, in-line heater, in-line temperature sensors, in-line DO sensors, in-line sensors for the presence of a medium, a cold store for storing and supplying reagents necessary for the implementation of biotechnological processes.

In some embodiments, the implementation of the control tower is connected to the gas station for the preparation and supply of gases necessary for the implementation of processes.

BRIEF DESCRIPTION OF THE DRAWINGS The objects, features and advantages of the present invention will become more apparent upon reference to the following description and the accompanying drawings, which show:

Fig. 1, Fig. 2 - Volumetric image of an asymmetric bioreactor according to the present invention,

Fig. 3 - Cross section of an asymmetric bioreactor,

Fig. 4 - Control tower with a pendulum mechanism, which is designed to hold and rock the asymmetric bioreactor,

Fig. 5 - An example of the layout and internal equipment of the control tower,

Fig. 6 - Fig. 8 - Asymmetric bioreactor system,

Fig. 9 -11 - The location of the ports of the asymmetric bioreactor,

Fig. 12 - Schematic representation of the operation of the asymmetric bioreactor system.

Detailed description of the invention

The present invention proposes an asymmetric conical bioreactor system for carrying out biotechnological processes occurring in the liquid phase during the cultivation of cells, microorganisms and viruses, which includes a multi-port sealed rigid vessel in the form of an asymmetric truncated cone with a rectangular trapezoid section with a smaller base at the bottom of the vessel and a support - a pendulum mechanism designed to hold the vessel and create a customizable and smooth movement (rolling) of the bioreactor relative to the vertical axis during the biotechnological process.

Typically, the support structure (pendulum) and the container (vessel) are designed to be used together. It is contemplated that a variety of vessels may be used in the pendulum mechanism, provided that they can be installed in the pendulum mechanism.

The elements of the design of the bioreactor and the pendulum support mechanism are detailed below.

The Asymmetric Conical Bioreactor is a new generation of unique single-use reactors with a smooth rocking motion. It is a scalable system with a working range from 15 ml to 50 liters. The operating range in this application refers to the range of the volume of liquids that can be handled in the reactor while maintaining full functionality (heating, contact with sensors, oxygenation of the environment, and so on). The device is designed for single use and especially considers the characteristics of cell cultures that are sensitive to sudden movements, which does not limit its other applications. The advantage of a specific reactor over reusable bioreactors is that maintenance and operation costs are reduced by eliminating the following operations:

- sterilization. Eliminates the need to have a fleet of sterilizers, eliminates the cost of equipment maintenance, certification and verification;

- cleaning. Eliminates the need to have a fleet of sterilizers, eliminates the cost of equipment maintenance, certification and verification;

- reduction of time and labor costs for regular maintenance in full compliance with the requirements;

- reduction of requirements for personnel qualification and costs for its training;

- reducing the risk of cross-contamination.

Embodiments of the bioreactor design disclosed herein according to the present invention have an operating range of 15 ml to 500 ml. The scalability of the system is achieved by configuring the inner surface of the culture vessel in the form of an asymmetric truncated cone, which has a rectangular trapezoid in cross section with a smaller base at the bottom of the vessel. The capacity of the bioreactor vessel may have a typical height to volume ratio of about 0.06-1 cm/L.

Fig. 1-3 are a three-dimensional view of a bioreactor 100 according to the present invention. The bioreactor 100 has a rigid vessel 110 with a wall 101 defining its cavity (internal volume of the vessel). The inner surface of the vessel 110 is made in the form of an asymmetric truncated cone and has a rectangular trapezoid in cross section with a smaller base at the bottom of the vessel. The outer surface of the vessel 110 has a predominantly vertical cylindrical shape with a beveled side wall.

Vessel 110 is sealed with lids on both ends: bottom lid 130 and top lid 120. Top lid 120 is connected to walls 101 of vessel 110 via flange 121. vessel weight - up to 8 kg. However, the above example in no way limits the overall dimensions of the structure of the vessel 110 of the bioreactor 100.

In some embodiments, at least one membrane is installed within the vessel 110, which divides the internal volume of the vessel into at least two spaces. A large area is used for growing cells, it contains a medium with cells and gases. In this case, liquid nutrient media are supplied through a smaller area (lower area), in which accumulation of gases is possible. In a smaller area, there is a drain valve with an extension to remove the air bubble from this space.

In the embodiment of the invention shown in FIG. 4 shows the use of two membranes 140, 150, one of which 140 is installed between the top cover 120 and the flange 121, and the second 150 is placed against the stops of the walls of the vessel 110 in the lower part of the vessel, closer to the bottom cover 130. In this case, both membranes 140, 150 installed parallel to each other and perpendicular to the longitudinal axis of the vessel 110.

Thus, the inner space of the vessel 110 of the bioreactor 100, bounded by the inner surface of the vessel, is divided into three chambers (Fig. 3) by membranes 140, 150, which are installed horizontally and form an upper space 161 bounded by the inner surface of the upper cover 120 and the first membrane 140, the lower a space 163 bounded by the inner surface of the bottom cover 130 and the second membrane 140, an intermembrane space 162 bounded by the first 140 and the second membrane 150.

Membranes can have different characteristics, material design and functionality depending on the requirements of the process. The membranes are replaceable and recyclable.

In some embodiments, a filter cartridge is installed inside the vessel.

The space 161 above the first membrane 140, the upper space, is used for ports to drain liquids or air through the cell retaining membrane 140.

The intermembrane space 162 forms a space into which it is possible to freely enter and remove reagents, but due to the size of the pores of the membranes, cells cannot leave this space. The membranes 140, 150 allow cultured materials to be retained in the intermembrane space and media can be introduced and withdrawn before or after the membranes, or reagents can be introduced and samples taken directly from the intermembrane space. Thus, cells are not lost during the perfusion of liquids in the reactor.

Vessel 110, depending on the requirements of the process and the desire of the end user, can be equipped with various sensors for controlling and monitoring the biotechnological process. Typically, reusable bioreactor systems use standard optical, capacitive, galvanic, and polarographic sensors to monitor the biotechnological process. The bioreactor 100 according to the present invention is equipped with probes 170, 180 - disposable or reusable sensors for monitoring the biotechnological process, selected from: a dissolved oxygen sensor DO, a pH sensor.

These sensors 170, 180 can be installed in the vessel 110 of the bioreactor 100 as standard or optional. Standard configuration of the sensors 170, 180 implies that the sensors in some embodiments are pre-installed. To install sensors for monitoring the biotechnological process, at least two mounting holes 171, 181 are placed on the outer surface of the vessel (Fig. 3). Moreover, the holes 171, 181 are placed in such a way that when the sensors 170, 180 are installed, they are completely immersed in the cultivated medium at any volume of liquid from the operating range. Sensors 180 are installed in mounting holes 171, 181 using push-in fittings 172, 182. In the embodiment shown in FIG. 3, ports 173, 183 are installed in the mounting holes 171, 181 in order to accommodate sensors for monitoring the biotechnological process, and the holes 171,181 are located on the side of the beveled side wall of the vessel 110 of the bioreactor 100 closer to the bottom cover 130 of the vessel. Sensors for monitoring the biotechnological process are installed in ports 173, 183 using fittings 172, 182 so that they are completely immersed in the cultivated medium at any volume of medium from the operating range.

In addition to the above sensors selected from: DO dissolved oxygen sensor, pH acid-base balance sensor, the bioreactor is equipped with disposable sensors selected from: glucose sensor, lactate sensor, temperature sensor, liquid level sensor, pressure sensor, biomass sensor. To install disposable sensors in the reactor, plugs and push-in fittings are provided. Thus, for example, in the embodiments of the invention depicted in FIG. 2-3, a pressure sensor 122 is provided in the flange in contact with the top cover 120.

The vessel surface, top cover 120, bottom cover 130, and flange 121 have mounting holes 190 for mounting ports. Each port 190 corresponds to a port 191. The ports 191 can be of various geometries and functions and be selected according to the requirements of the biotechnological process. The minimum required ports are 191: gas inlet port, gas outlet port, sampling port, culture inlet port, reagent inlet port, culture collection port, waste outlet port, membrane air bubble outlet port. Optionally, additional gas ports and ports for the introduction of activators and other reagents, alternative ports for the collection of grown crops and alternative ports can be used.

In some embodiments, a sensor, such as a temperature sensor, may be installed at port 191.

191 port connectors with landing holes have the same thread, which allows you to change the connection type to the appropriate one at any time. The use of sensor connectors and ports, such as luer locks, provide a reliable connection, allowing biotechnological processes to run for as long as required. Moreover, the design of the proposed bioreactor vessel makes it possible to carry out all stages, for example, the process of obtaining CAR-T cells, in one vessel. This process includes the activation of T lymphocytes, their transduction with a lentiviral vector, and expansion (increase in number).

The complex shape of the inner surface of the vessel 110 of the bioreactor 100 allows you to start the process of cultivation and operation of the medium with minimal volumes - it saves reagents and at the same time allows you to create a sufficient height of the liquid column to accommodate the sensors and maintain their functioning. Sensors 170, 180 for monitoring the biotechnological process will always be immersed in the environment for the purpose of monitoring the process.

In an embodiment of the invention, when at least one membrane is installed inside the vessel 110, a landing hole 131 is made on the bottom cover 130 of the vessel, to which port 132 corresponds. air bubble, in particular, in the lower space 163 of the vessel 110 of the bioreactor, limited by the inner surface of the lower cover 130 and the second membrane 150. In the process of filtration under the lower membrane 150 of the bioreactor, an air bubble may form, preventing the continuous and uniform supply / removal of liquids. The extension 133 allows the air bubble that accumulates under the second membrane 150 to be removed. membrane or create a burst air bubble, which in turn creates a strong bubbling and can damage the cell culture.

During the operation of an asymmetric conical bioreactor, medium and other liquids are fed into it through flexible tubes at a user-defined speed, which immediately after entering the bioreactor pass through the first filter membrane 140 (Fig. 3).

The location of the ports 191 in relation to the top or bottom of the vessel is not critical and can be anything. Their location in height is important, but not their angle. The ports 191 may be located in the intermembrane space 162, in the upper space 161 bounded by the inner surface of the upper cover 120 and the first membrane 140, in the lower space 163 bounded by the inner surface of the lower cover 130 and the second membrane 150. Ports 191 are located in such a way that they can enter or divert media from the membrane-bound or single-membrane spaces of the bioreactor. The choice of ports 191 and their location is determined by the requirement of a particular process.

In some embodiments, the wall 101 of the multiport rigid vessel 110 for culturing cells, microorganisms, and viruses can be made from a variety of biocompatible materials. For example, stainless steel, polycarbonate, PTFE or polysulfone. In other embodiments, metallic materials, ceramics, or composites are used.

In other embodiments, stainless steel is used, alone or in combination with glass. For example, in the manufacture of the wall 101 of the vessel of the bioreactor 100 from stainless steel (Fig. 2), a bar 102 and a sight glass 103 are provided for visual assessment of the environment inside the bioreactor 100. In this case, the covers 120, 130 are made of fluoroplastic.

In some embodiments, vessel 110 is made of Teflon and polysulfone (high performance thermoplastics) (vessel 110 is polysulfone and lid 120, 130 is Teflon) or glass. In some embodiments, the vessel 110 is made from polycarbonate (the vessel and components are polycarbonate). The choice of materials from which one or another part of the bioreactor is made is not limited to these materials. In general, the list of vessel materials is limited only by the biocompatibility of the materials.

For the manufacture of a steel vessel, the following manufacturing methods can be used: milling, electroerosion machining, turning operations. For the manufacture of plastic vessels, injection molding, milling, turning operations, and 3D printing are used.

For the implementation of the technological process, heating and constant maintenance of the temperature of the contents of the bioreactor are necessary, therefore heating of the bioreactor vessel is provided. As a means of heating to maintain the temperature of the cultivation of the bioreactor, in one of the embodiments, a heating jacket can be used, which is fixed on the outer surface of the bioreactor vessel (on the body of the bioreactor) using laces, a rigid casing that repeats the shape of the bioreactor vessel, or other available method of fastening. The heating jacket can simply be put on the bioreactor vessel due to the elasticity of the casing materials and fixed on the vessel with a bolt or nylon ties.

In another embodiment, the heating means is a heating cartridge inserted directly into the interior of the vessel instead of a plug located in the flange.

In another embodiment, the heating means is a heating cartridge for heating the walls of the vessel, installed in special holes in the vessel body of the bioreactor. For example, installation of the specified cartridge is possible in the embodiment when the material of the bioreactor vessel is stainless steel and the vessel has at least one hole in the vessel wall.

In another embodiment, the heating means is a heater built into the wall of the bioreactor vessel. This embodiment is possible in the case of using a plastic body of the bioreactor.

It is also possible to implement the heating means in the form of a heat-conducting sprayed layer on the body of the bioreactor on the inner or outer surface of the vessel. Wires are soldered to the sprayed layer and brought out. The layer can be deposited using known technologies, for example, using vacuum metal deposition technology. On the outer surface of the body of the bioreactor, a layer of thermal insulation is applied over the sprayed layer. On the inner surface of the body of the bioreactor, a layer of inert biocompatible material is applied over the sprayed layer.

In another embodiment, the contents of the bioreactor vessel are heated to the required temperature using a flow heating system for the fluid supplied to the bioreactor.

Bioreactor 100 can operate in automatic, semi-automatic and manual modes.

In the case of full manual control, all processes including tubing, pumping liquids/gases (using syringes or self-contained peristaltic liquid pumps and a pressure calibrated compressed air/carbon dioxide/oxygen/nitrogen source), pumping (using a shaker or similar device), harvesting and disposal of waste is carried out manually or using user equipment.

In the case of automatic or semi-automatic operation, all processes, except for installation, are carried out automatically using the bioreactor system.

The second aspect of the present invention relates to a bioreactor system, the design and functionality of which are described in detail below.

Bioreactor system 200 for culturing cells, microorganisms, and viruses includes a bioreactor according to any embodiment described above and a control tower 210 (FIG. 4) consisting of a pendulum support mechanism 220 (FIG. 5) configured to hold, rotate, and swing a bioreactor vessel 100 vertically installed in it relative to its vertical axis during the biotechnological process, an electrical connection panel 230, power supplies 214, 215, a control unit 250, a control panel 216. The control tower 210 modularly and flexibly, with the possibility of expansion, houses all the necessary electronics to ensure the operation of the system.

The pendulum mechanism 220 (Fig. 4, 5) for bioreactors is presented in the form of a clamping collar 221, mounted on the motor shaft 222, which is mounted on a rack 223, rigidly fixed to the base plate 224. In the preferred embodiment, a stepper is used as the motor 222 geared motor with a torque of at least 10 N/m. In a non-limiting embodiment, a servo with enough torque to drive the vessel can be used. Rack 223 may be made in the form of a square tube. The material of the column 223 and the base plate may be stainless steel. Racks 223 are welded together in a frame 225, and a support plate 224 is screwed to the frame. An engine 222 is installed in the frame 225, connected to the bioreactor fastening device 226 (with a clamping collar 221) by screws. To track the angle of inclination of the bioreactor vessel 100, an accelerometer 227 is mounted on the clamp collar 221 (FIG. 4). The control tower body 211, made of plastic, is bolted to the frame. In the seats on the housing 211 installed fans 212 bolted. Inside the housing 212, on the guide mounting rails 213, fixed on the base plate 224, the control unit 214 and the power supply unit 215 are installed. The electrical components are switched among themselves. The control tower 210 is also equipped with peristaltic pumps 216 for supplying reagents to the bioreactor vessel 100. Thus, the entire mechanism, together with the two peristaltic pumps 216, the cooling fans 212 and the control unit 214, is enclosed in a plastic case 211.

In one of the non-limiting examples of the invention, the support-pendulum mechanism has: weight 40 kg, overall dimensions of the mechanism are 500 x 360 x 600 mm, the inner diameter of the clamping collar is 96 mm, the maximum length from the bottom of the bioreactor to the clamping collar of the pendulum mechanism is up to 300 mm, the maximum the length from the clamping collar of the pendulum mechanism to the top fittings of the bioreactor is up to 300 mm. In this case, it is possible to use a bioreactor with a maximum weight of up to 10 kg with this pendulum mechanism.

The pendulum mechanism can be used in the development of technological processes in the field of biopharmaceuticals and immunotherapy to rock the disposable bioreactor fixed on it at a speed that is safe for sensitive cell cultures (for example, mammalian cells, human cells), to ensure thorough mixing of the liquids in the bioreactor.

The bioreactor is installed in the clamping collar of the pendulum mechanism driven by a stepper motor. The clamping collar oscillates cyclically with a pendulum motion, for example, at a speed of 57 minutes, tilting the bioreactor at a predetermined angle selected by the user (the value can also be selected/adjusted during operation), depending on the amount of content, the type of bioreactor or the requirements of the technological process.

Examples of maximum tilt angles depending on the filling of the reactor:

- 83° from the vertical axis (when filling up to 500 ml);

- 73° from the vertical axis (when filling up to 300 ml);

- 61° from the vertical axis (when filling up to 100 ml);

- 23° from the vertical axis (when filling up to 15 ml).

The speed of movement of the clamping yoke in which the bioreactor is installed is determined by the current biotechnological process and the selected type of bioreactor. For example, the speed can reach up to 400 rpm.

All necessary parameters, such as swing speed, swing duration and tilt angle, are selected by the operator using the control panel 209, made in the form of a touch display connected to the control unit 250 and located on the body of the control tower 211.

The control unit 250 is configured to execute commands from the peripheral computing devices and/or the control panel 209 integrated with the pendulum mechanism 220 to set the current parameters of the biotechnological process. Also, the control unit 250 should additionally and preferably be capable of:

receive process trend data from disposable or reusable sensors to obtain measurements of the physical, chemical or biological properties of the contents of the vessel during the biotechnological process,

adjust the parameters of the biotechnological process depending on the measurements obtained by applying control signals to the mechanisms for supplying gas 260, culture media to the vessel 110 of the bioreactor 100, configured to deliver gas, culture media to the vessel 110 of the bioreactor 100 during events in response to the control signal, to the means heating, configured to change and maintain the temperature in the vessel 110 of the bioreactor 100 in response to the control signal, and to the pendulum mechanism 220, configured to change the angle of inclination in response to the control signal.

The control tower 210, mounting plate 270, gas supply mechanism 260, and reactor 100 are preferably connected to various instruments in the operation of the present invention, forming a system 200 (FIG. 6) for automatic or semi-automatic operation of the present invention. For example, the control tower 210 is preferably connected to the gas supply mechanism 260 to control the flow rate of a gas or combination of gases. This device is preferably connected to a gas cylinder or line from a suitable air supply and controls the flow of air or other gas in the pipe.

The control unit 250 is also preferably connected to a thermometer, which is preferably present in the environment of the bioreactor vessel, but more preferably not in the bioreactor vessel. The control unit 250 should also additionally and preferably be able to control a temperature control mechanism such as a heating means and/or a cooler.

The control unit 250 is additionally and preferably connected to a device that controls the flow of the medium and/or other nutrients from the nutrient/media container to the bioreactor vessel container.

The control unit 250 is preferably connected to at least one channel of the bioreactor vessel and more preferably at least to the collection channel of the bioreactor vessel. The control unit 250 may additionally control an automated sampling and/or collection means to remove a portion of the contents of the vessel vessel of the bioreactor from an additional sampling channel, for inspection and/or collection (not shown). The control unit can also be additionally connected to the analyzer to analyze the selected part of the content, for example, to provide feedback for the operation of the control unit.

The control unit 250 may be configured to control battery performance and/or temperature, the amount and time of entry of a gas or combination of gases into the container, the amount of gas and the time it leaves the container, the amount and time of addition of at least one material (nutrient , culture medium, etc.) and/or amount of light. The control unit is capable of detecting the amount of waste received.

In some embodiments, the control unit is configured to take into account the dynamics of cell growth in real time, and the control panel is configured to configure graphical and 3D visualization of processes; user task configuration of processes; saving/loading data; remote monitoring of the experiment.

The connection of the control unit 250 to such elements as reusable or disposable sensors, shutoff valves, mechanisms for supplying reagents to the bioreactor vessel, a stepper motor of the oscillating mechanism and other equipment necessary to ensure the biotechnological process is carried out through the electrical connection panel.

The control tower is connected via a control unit and an electrical connection panel to the gas station for the preparation and supply of gases necessary for the processes.

To connect additional equipment, the control tower 210 is equipped with a mounting plate 270. Mounting plate 270 is installed so that it is located in the area of the bioreactor vessel. Mounting plate 270 can accommodate the equipment required for the process, selected from: piping, pinch valves, fittings, oxygenator, bubble traps, in-line heater, in-line temperature sensors, in-line DO sensors, in-line fluid presence sensors, and other components, necessary for the implementation of biotechnological processes.

The power supply unit is configured to power the pendulum mechanism, peristaltic pumps, sensors, display, light and sound indication, fans, control device, pinch valves, fluid flow heater, heating means.

As a touch display in the preferred embodiment of the invention, a capacitive HDMI display with dimensions of 11.6'', 1920×1080 can be used. Display controller: RTD2556. Current consumption: 1 A. Supply voltage: 12 V. Touch panel controller: ILI2511. Touchscreen interface: USB

The user has the ability, for example, to set the parameters of the biotechnological process using the menu displayed on the touch screen of the pendulum mechanism (located on the body) or, more conveniently, using a web application.

In the first case, after entering data on the touch screen, commands from the control panel are transmitted to the computing module, which, in turn, issues commands to the control unit.

In the latter case, when setting the current parameters of the biotechnological process through a web application, the web application for the task is launched on peripheral computing devices, which can be selected as a laptop, personal computer, mobile phone. The peripheral computing devices are configured to transmit data to the computing module, which issues commands to the control unit.

On the body of the control tower, LED indicators can also be located, which are electrically connected to the control unit and the operator about the status of the process. For example, LED indicators can signal at least one of the following events:

- about an emergency;

- on completion of the process;

- about the deviation of the process indicators from the required ones;

- about the readiness of the equipment to start the process

- about finding the system in operation.

The heating temperature and its maintenance at the level set by the user is carried out automatically by means of a control panel integrated with a pendulum mechanism used to rock the bioreactor and control all the necessary parameters of the technological process. In addition, it is also possible to control using a PC/tablet/mobile phone via the web interface.

The display of the control panel/personal computer/tablet/mobile phone graphically displays all parameters of the technological process (including temperature): maximum speed and maximum angle of inclination of the bioreactor vessel from the vertical axis of the pendulum mechanism, the volume and temperature of the supplied reagents and the speed of their supply , vessel content heating temperature, vessel pressure, pH, DO, Glu, Lac, Biomass, peristaltic pump flow and others.

In addition, the control unit is configured to notify the operator via the control panel (via the touch display integrated into the control tower housing, or through the display of the web application) about the need to perform certain actions and, if necessary, issue an alert in case the instrument readings go beyond the limits. specified ranges.

The control panel 209 includes data input and recording means for controlling the culture conditions in the vessel and controlling the fluid distribution valves in sequence. Automatic registration and recording of all events and actions with the system throughout the entire process, including sensor readings. Access to all data can be stored in the cloud, which can be accessed remotely from any device, which allows you to monitor current processes in real time.

The procedure for operating the bioreactor system for culturing cells, microorganisms and viruses is as follows.

The bioreactor with a length not exceeding the specified one is fixed in the clamping collar of the pendulum mechanism. To do this, the screws are unscrewed from the lugs of the clamping collar and the unscrewed part of the clamping collar is removed. The bioreactor vessel is placed in the clamping yoke. Install the previously unscrewed part of the clamping collar in place, install and tighten the screws.

The heating means and pinch valve supports are then installed. The pinch valves are mounted on a mounting plate, which is screwed to the control tower body.

Through the distribution system, which is deployed on the mounting plate 270, flexible tubes 275 and wires are pulled. Next, attach the flexible tubing to the bioreactor vessel in accordance with the labels and color identification.

Install network connectors (Ethernet, LAN, etc.) in accordance with the labels and color identification: the device must be powered from a 220 V network and connected to the Internet. Connect the external power supply to a grounded outlet.

With the help of the control panel (touch screen or web application), the required parameters of the technological process (swing speed, swing duration, tilt angle) are set and the technological process is started, which is also controlled through the control panel.

The bioreactor vessel swings smoothly at a predetermined speed for a predetermined time selected by the operator on the touch screen of the pendulum mechanism located on the body, or using a web application. Rocking causes movement of the cell culture medium in the bioreactor. This movement ensures thorough mixing of the cell suspension, medium, and other added fluids at a rate that is safe for sensitive cells, and also provides the necessary level of mass transfer.

The pendulum mechanism stops after a predetermined time or during certain operations, such as adding reagents, removing waste or withdrawing product. “STOP” appears on the display of the control panel and the device will beep repeatedly. The operation can be stopped at any time before the set time has elapsed by pressing the STOP button. The pendulum mechanism will assume a vertical position and stop.

The device is designed to work in rooms with low air temperature, as well as in closed laboratory rooms at room temperature from +4°С to +40°С in non-condensing conditions and at a maximum relative humidity of 80% at temperatures up to 31°С, linear decreasing to 50% at 40°C.

To remove the air bubble that is formed under the membrane 150 in the area of the lower space 183 of the bioreactor vessel, the bioreactor is tilted at a small angle from the vertical axis while simultaneously supplying liquid.

Withdrawal is performed as follows:

(a) the bioreactor vessel is tilted so that the air bubble moves closer to the extension 133;

(b) through port 7 (see Fig. 9, 11) using pump 216, the medium is supplied to the bioreactor;

(c) through the extension 133 using another pump (not shown in the drawings), the medium and air are pumped out of the bioreactor 100 into the waste container.

In FIG. 6-8 shows an example of an assembled bioreactor system for culturing cells, microorganisms and viruses, consisting of a control tower, an asymmetric conical bioreactor and peripheral equipment:

- mounting board 270, which accommodates additional equipment that is required to ensure the process;

- pinch valve 271, quantity and location determined by the requirement of the process;

- oxygenator 272 for enriching the loaded medium with oxygen;

- flow heater 273 for heating the medium before loading into the heater;

- flow temperature sensor 274, to control the temperature of the loaded medium;

- gas station 260, for the preparation and supply of gases necessary for the process.

Examples of the use of this device for culturing CAR-T cells and obtaining HEK293 cells are shown below (Fig. 9-12).

CAR-T PROCESS

1. Plant the cells through port #6 with a luer lock syringe.

2. Add the required volume (for ImmunoCult CD3/CD28 - 25 µl of activator per 1 ml of medium) and 100 U/ml of IL2 through port #11 using a syringe.

3. Turn on a gentle incline of 5 degrees in 1 minute.

4. After activation has passed (24-72 hours), tilt the reactor to a horizontal position towards the product collection/waste collection port at a speed of 5 degrees per minute.

5. Open the drain valve (port #13) and pressurize inside the reactor with N2 through port #15. In this case, the exact pressure parameters should be determined experimentally, but in the range of 1-2 bar.

6. When all of the medium has been removed, close the valve and reduce the pressure.

7. Fill the reactor with preheated (37°C) Wash Buffer (HPBS) to half the volume of the reactor through port #7 at a rate of 10 ml/min.

8. Repeat steps 5-6.

9. Add the volume of prewarmed fresh medium (AIM-V with IL-2) at the volume required to achieve a cell density of 5*10 ⁶ /ml through port #8 at a rate of 10 ml/min.

10. Return the reactor to a vertical position at a speed of 5 degrees in 1 minute.

11. Add lentivirus and transduction agent through appropriate ports #9 and #10 using a syringe. The volume of lentivirus should be calculated from the virus titer and the desired MOI. Volume of hexadimethrin bromide 6 μg/ml (transduction agent).

12. Turn on a gentle tilt at a speed of 5 degrees for 1 minute.

13. After transduction (16 hours), repeat steps 5-10.

14. Turn on a gentle tilt of 5 degrees per minute and add fresh medium (AIM-V with IL-2) through port #8 at 10 ml/min to double it when the cell density increases by 2*10 ⁶ /ml.

15. When the desired number of cells (according to the cell count in the sample) is reached, repeat steps 5-8.

16. Fill the reactor with buffer for packaging the finished product through port #4 (or medium through port #8 if QC testing is not performed according to GMP standards in the current experiment) with the volume necessary to obtain the final cell concentration of the product (5-10*10 ⁶ / ml) at a rate of 10 ml/min.

17. Open port #12 of collection valve 1 and pressurize inside the reactor with N2 through port #15.

Note: the exact pressure parameters should be determined experimentally, but in the range of 1-2 bar.

18. When the cell collection is complete (look through the glass window), close the valve and reduce the pressure. The process of obtaining CAR-T cells is over.

Samples are taken through port #3 or #5 using a secure vacutainer system (3-5 ml) twice daily to check cell viability, number and transduction efficiency throughout the process.

HEK293 PROCESS

1. Fill the reactor with 150 ml prewarmed (37°C) medium (MEM containing 10% FBS (heat inactivated), 100 units/ml penicillin/streptomycin and 2 mM L-glutamine) through port #8 at a rate of 20 ml/min .

2. Inject 300*10 ⁶ cells through port #6 using a luer lock syringe.

3. Turn on a gentle incline of 15 degrees for 1 minute.

4. Grow the cells and add fresh medium through port #8 at a rate of 20 ml/min to double it when the cell density rises to 4*10 ⁶ / ml. Continue until the maximum reactor volume (450 ml) is reached.

5. Tilt the reactor to the horizontal to collect/waste at a rate of 15 degrees per 1 minute.

6. Open the drain valve (port #13) and pressurize inside the reactor with N2 through port #15.

7. When all of the medium has been removed, close the valve and reduce the pressure.

8. Fill the reactor with preheated (37°C) Wash Buffer (HPBS) to half the volume of the reactor through port #7 at a rate of 20 ml/min.

9. Repeat steps 6-7.

10. Add prewarmed (37°C) medium through port #8 at a rate of 20 ml/min in a volume equal to that extracted in step 7.

11. Return the reactor to a vertical position at a speed of 15 degrees in 1 minute.

12. Add transfection mix through port #9 using a syringe.

13. When transfection is complete (6 hours), repeat steps 5-11.

14. Turn on a gentle tilt of 15 degrees for 1 minute for 48-72 hours to get lentivirus.

15. When the lentivirus is ready (48-72 hours), tilt the reactor to a horizontal position at a rate of 15 degrees per minute.

16. Open port #14 of valve 2 to collect the product, pressurize the inside of the reactor with N2 through port #15 and collect the medium with lentivirus.

17. You can then close the valve and repeat steps 7-8 and then collect the cells through collection port #12. The HEK293 process is over.

Samples are taken through port #3 or #5 using a secure vacutainer system (3-5 ml) twice daily to check viability, cell count throughout the process.

Although the present patent application refers to what is defined in the appended claims below, it is important to note that this patent application contains the basis for the formulation of other inventions, which may, for example, be claimed as the subject of the amended claims of the present application or as the subject of the claims in the emphasis. application. Such an object may be characterized by any feature or combination of features described herein.

Claims (60)

1. Bioreactor for the implementation of biotechnological processes occurring in the liquid phase during the cultivation of cells, microorganisms and viruses, including a hermetically sealed rigid culture vessel comprising a top and bottom lid, the top lid being flanged to the vessel, at the same time, the inner surface of the vessel is made in the form of a truncated cone and has a rectangular trapezium in cross section with a smaller base at the bottom of the vessel, and on the outer surface of the vessel there are at least two mounting holes for installing sensors for monitoring the biotechnological process in such a way that when installing the sensors they are completely immersed in the cultivated medium when filling the vessel with it, moreover, on the surfaces of the vessel, the top cover, the bottom cover and the flange, there are seats for installing at least one gas inlet port, a gas outlet port, at least one port for sampling, a port for introducing cultures, at least one port for introducing reagents, at least one port for the collection of cultivated crops, a port for waste disposal. 2. Bioreactor for the implementation of biotechnological processes occurring in the liquid phase during the cultivation of cells, microorganisms and viruses, including a hermetically sealed rigid culture vessel comprising a top and bottom lid, the top lid being flanged to the vessel, at the same time, the inner surface of the vessel is made in the form of a truncated cone and has a rectangular trapezium in cross section with a smaller base at the bottom of the vessel, on the outer surface of the vessel there are at least two mounting holes for installing sensors for monitoring the biotechnological process in such a way that when installing the sensors they are completely immersed in the cultivated medium when filling the vessel with it, moreover, on the surfaces of the vessel, the top cover, the bottom cover and the flange, there are seats for installing at least one gas inlet port, a gas outlet port, at least one port for sampling, a port for introducing cultures, at least one port for introducing reagents, at least one port for collecting grown crops, a port for waste disposal, a port for removing an air bubble from under the membrane, at the same time, at least one membrane is installed inside the vessel, which divides the internal volume of the vessel into at least two spaces, one of which is the region of the gas accumulation space, and in the area of the gas accumulation space there is a drain valve with an extension to remove the air bubble from this space. 3. The bioreactor according to claim 2, in which the vessel has a top cover and a bottom cover, while the internal space of the vessel, limited by the internal surface of the vessel, is divided into three chambers by membranes, which form an upper space bounded by the inner surface of the top cover and the first membrane, a bottom space defined by the inner surface of the bottom cover and the second membrane, intermembrane space bounded by the first and second membranes. 4. The bioreactor according to claim. 1 or 2, in which the outer surface of the vessel has a predominantly vertical cylindrical shape with a beveled side wall, the vessel is sealed with lids on both ends. 5. The bioreactor according to claim 4, in which the vessel and lids have mounting holes for installing ports, the geometry and functionality of which is selected in accordance with the requirements of the biotechnological process. 6. Bioreactor according to claim. 1 or 2, in which the sensors for monitoring the biotechnological process are disposable or reusable. 7. The bioreactor according to claim 6, in which the sensors for monitoring the biotechnological process are selected from: a dissolved oxygen sensor DO, an acid-base balance sensor PH. 8. The bioreactor according to claim 7, which additionally includes disposable sensors placed in the vessel openings and selected from: a glucose sensor, a lactate sensor, a temperature sensor, a liquid level sensor, a pressure sensor, a biomass sensor. 9. Bioreactor according to claim. 1 or 2, in which the sensors for monitoring the biotechnological process are pre-installed. 10. Bioreactor according to claim 1 or 2, in which the internal volume of the vessel is from about 15 ml to 50,000 ml. 11. The bioreactor according to claim. 1 or 2, in which each landing hole corresponds to a port, while the ports are selected from the group: a gas inlet port, a gas outlet port, a sampling port, a port for introducing cultures, a port for introducing reagents, a port for collection of cultivated crops, a port for waste disposal, a port for diverting an air bubble from under the membrane. 12. Bioreactor according to claim. 1 or 2, in which the material of the vessel and/or lids is a biocompatible material or a combination of biocompatible materials, while the biocompatible material is selected from the group: glass, polyethylene, stainless steel, polycarbonate, polysulfone. 13. Bioreactor system for culturing cells, microorganisms and viruses, including bioreactor according to any one of paragraphs. 1-12 with heating means and a control tower equipped with a pendulum support mechanism configured to hold, rotate and rock the bioreactor vessel vertically installed in it relative to its vertical axis during the biotechnological process, electrical connection panel power supply, control unit configured execute commands from peripheral computing devices and / or a control panel integrated with a pendulum mechanism to set the current parameters of the biotechnological process, receive process trend data from disposable or reusable sensors to obtain measurements of the physical, chemical or biological properties of the contents of the vessel during the biotechnological process, adjust the parameters of the biotechnological process depending on the measurements obtained by applying control signals on mechanisms for supplying gas, nutrient media to the bioreactor vessel, configured to deliver gas, nutrient media to the bioreactor vessel during events in response to a control signal, to a heating means configured to change and maintain the temperature in the bioreactor vessel in response to a control signal, and a pendulum mechanism configured to change the angle of inclination in response to a control signal. 14. The system according to claim 13, in which the pendulum mechanism is made in the form of a clamping collar fixed on the stepper motor shaft, mounted on a stand fixed on the base plate. 15. The system according to claim 13, in which the current parameters of the biotechnological process are selected from: the maximum speed and the maximum angle of inclination of the bioreactor vessel from the vertical axis of the support-pendulum mechanism, the volume and temperature of the supplied reagents and their supply rate, the heating temperature of the contents of the vessel, the pressure in Vessel, pH, DO, Glu, Lac, Biomass. 16. The system of claim. 13, in which the control tower is equipped with peristaltic pumps for supplying reagents to the vessel of the bioreactor. 17. The system according to claim 13, in which the control panel is located on the body of the control tower and is made in the form of a touch display connected to the control unit. 18. The system of claim. 17, in which the control panel is configured to transmit data to the computing module, which issues commands to the control unit. 19. The system according to claim 13, in which peripheral computing devices are selected from: a laptop, a personal computer, a mobile phone, and are configured to run a web application on them to set the current parameters of the biotechnological process. 20. The system of claim 19, wherein the computing peripherals are configured to communicate data to a computing module that issues commands to a control unit. 21. The system according to claim 13, in which the electrical connection panel is configured to connect the pendulum mechanism, reusable or disposable sensors, valves, mechanisms for supplying reagents to the bioreactor vessel, equipment necessary to ensure the biotechnological process. 22. The system according to claim 13, in which the power supply unit is configured to power the pendulum mechanism, peristaltic pumps, sensors, display, light and sound indication, fans, control device, pinch valves, fluid flow heater, heating means. 23. The system according to claim 13, in which LED indicators are located on the control tower body, associated with the control unit and signaling at least one of the events: - emergency situation; - completion of the process; - deviation of process indicators from the required ones; - readiness of the equipment to start the process; - finding the system in operation. 24. The system of claim. 13, in which the heating means is a heating jacket, which is put on the outside of the outer surface of the vessel to maintain the temperature of the cultivation of the bioreactor. 25. The system according to claim 13, in which the material of the bioreactor vessel is stainless steel, the vessel has at least one hole in the vessel wall for installing a heating cartridge, and the heating means is a heating cartridge for heating the walls of the vessel inserted into one of these holes. 26. The system of claim. 13, in which the means of heating is a heating cartridge inserted directly into the interior of the vessel. 27. The system of claim. 13, in which the means of heating is a built-in heater in the vessel body of the bioreactor. 28. The system according to claim 13, in which the heating means is a heat-conducting sprayed layer by vacuum deposition technology on the inner or outer surface of the vessel with wires soldered to the sprayed layer and brought out. 29. The system according to claim 13, which additionally includes a system for flow-through heating of the liquid supplied to the reactor to the required temperature. 30. The system according to claim 13, in which a mounting plate is installed on the control tower so that it is located in the area of the bioreactor vessel, for placing on it the equipment required to provide a process selected from: pipeline, pinch valves, fittings, oxygenator , bubble traps, in-line heater, in-line temperature sensors, in-line DO sensors, in-line media presence sensors, cold room for storage and supply of reagents required for biotechnological processes. 31. The system according to claim 13, in which the control tower is connected to a gas station for the preparation and supply of gases necessary for the implementation of processes.

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