Equipment for automatic cell expression of recombinant protein

Abstract

The invention discloses automatic recombinant protein cell expression equipment, which comprises a plurality of independent compartments, a sample treatment device, a cell culture device, a sample monitoring device, a compartment transfer device, a moving device and a sterilization device, a temperature and humidity monitoring and control device is arranged in the cell culture device; sterilization devices are arranged in the sample treatment device, the cell culture device, the sample monitoring device and the compartment transfer device; an air circulation device is arranged in the sample treatment device, the air circulation device comprises an air inlet device and an air outlet device, air outside the equipment enters the air inlet device through the air inlet, passes through the air outlet device and then is discharged out of the equipment through the air outlet, and one-way airflow from one side to the other side is formed in the equipment. The equipment can complete automatic operation of samples under the condition of no manual intervention, so that cross contamination and environmental pollution between processes are avoided, and the production efficiency and the product quality are improved.

Description

Technical Field

[0001] The invention belongs to the field of automated equipment, and in particular relates to an automated cell-expressing recombinant protein device. Background Art

[0002] Recombinant protein expression has become an important tool in modern biotechnology, with a wide range of applications in medicine, agriculture, and industry. In the biotechnology industry, the expression of recombinant proteins in cells is a crucial process. Traditional methods for expressing proteins in mammalian cells involve tedious and time-consuming steps such as transfection or transformation, expansion culture, and cell yield monitoring. In addition, current manual methods suffer from low yields, high variability, and contamination, and cannot optimize culture conditions more effectively. At present, there are relatively complete solutions for the automation of the cell culture process. For example, there is a semi-automatic culture method for the treatment of adherent cells (CN207727091U), but similar methods cannot achieve full process automation, nor can they detect and collect the expression products; for the treatment of suspended cells, there are large-scale culture technologies (Mei Jianguo, et al. "Large-scale culture technology of animal cells." Chinese Journal of Biotechnology 32.07 (2012): 127-132.), automatic cell culture devices (CN109642201A), and automated cell culture systems (CN106635790B). The existing culture devices are suitable for the culture of cells with confirmed protein expression, but cannot be used for the subsequent steps of optimizing and screening protein expression conditions after cell culture, and cannot meet current needs.

[0003] Regarding the process of cell expression of recombinant proteins, existing equipment cannot achieve full-process automation from cell culture to expression product detection and collection, especially in the optimization and screening of protein expression conditions, which still requires a lot of manual intervention. In addition, the existing technology is also insufficient in ensuring cleanliness during the culture process, and cannot ensure a stable sterile environment in multi-step operations. Therefore, it is particularly important to develop a device with high automation and cleanliness control. Through the clean control system of this device, it is possible to reduce manual intervention and effectively reduce external contamination during culture, expression and subsequent operations, ensuring the smooth progress of each link, while significantly improving the reliability of the protein expression process and the efficiency of condition optimization and screening. Summary of the invention

[0004] In order to optimize the protein expression process and reduce manual intervention, and to improve the reliability of the protein expression process and the efficiency of optimizing and screening related protein expression conditions, the present invention provides an automated cell-based recombinant protein expression device.

[0005] In a first aspect, the present invention provides a device for expressing a recombinant protein in a cell, the device comprising a plurality of independent compartments, wherein different compartments of the device are provided with a sample processing device 104, a cell culture device 105, a sample monitoring device 106, compartment transfer devices 205 and 206, and a sterilization device 311; the cell culture device 105 is provided with a temperature and humidity monitoring and control device 3012; the sample processing device 104, the cell culture device 105, the sample monitoring device 106, and the compartment transfer devices 205 and 206 are provided with There is a sterilization device 311; the sample processing device 104 is provided with an air circulation device, the air circulation device includes an air inlet device 301 and an air outlet device 302, the part of the air inlet device 301 connected to the device shell is provided with an air inlet 107, the part of the air outlet device 302 connected to the device shell is provided with an air outlet 108, the air outside the equipment enters the air inlet device 301 through the air inlet 107, and is discharged from the equipment through the air outlet 108 after passing through the air outlet device 302, forming a unidirectional airflow from one side to the other side in the equipment.

[0006] In some embodiments, the air inlet device 301 is provided with a filtering component, such as a HEPA filter.

[0007] In some embodiments, the sample processing device 104 is further provided with a sample preprocessing device, a sample adding device, a waste collection device and a consumable storage device; and / or,

[0008] The device also includes control system means.

[0009] In some embodiments, each of the compartments is provided with a sterilization device 311 .

[0010] In some embodiments, the compartment is a clean controlled environment or a low contamination risk environment.

[0011] Installing a sterilization device in the compartment can effectively kill or remove bacteria, viruses, fungi and other microorganisms in the compartment, reducing the risk of biological contamination in the compartment. At the same time, it can remove dust particles, harmful gases, etc. in the air, improve the air quality in the compartment, reduce the risk of particulate pollution, and thus make the compartment a clean controlled environment or a low-pollution risk environment.

[0012] In some embodiments, the sample pretreatment device is located in a separate compartment.

[0013] In some embodiments, the sample pretreatment device includes a cell culture fluid pretreatment device 201, a cell sample pretreatment device 202, a transfection sample pretreatment device 203, and a waste pretreatment device 204.

[0014] In some embodiments, the sample processing device 104, the cell culture device 105, the sample monitoring device 106, and the compartment transfer devices 205 and 206 are disposed in separate compartments.

[0015] The sample processing device 104 includes different experimental consumables areas 6003 and 6004 , a monitoring and transfection sample loading area 5006 , a cell culture fluid area 5003 , a cell sample area 5005 and a waste collection area 5007 .

[0016] In some embodiments, the consumables storage device includes storage areas 603 and 604 for different experimental consumables to ensure the classified placement and use of experimental consumables; the cell culture device 105 includes an area 601 for storing equipment and instruments related to cell culture; the sample monitoring device 106 includes an area 602 for storing monitoring instruments for monitoring cell growth and induction status.

[0017] The equipment and instruments related to cell culture include incubators, biosafety cabinets, liquid nitrogen tanks, freezers, filtration devices, etc.

[0018] The monitoring instruments used to monitor the growth and induction status of cells include optical microscope, confocal laser scanning microscope (CLSM), living cell imaging system, flow cytometer, cell counter, cell analyzer, bioluminescence and fluorescence imaging system, etc.

[0019] In some embodiments, the sample processing device 104 is provided with a sample loading device and a regional positioning system 401, and the regional positioning system 401 is equipped with a pipette head 4001. The sample loading device draws liquid from one area through the pipette head 4001 equipped with the regional positioning system 401 and adds it to the sample in another area.

[0020] In some embodiments, the sample processing device 104 and the cell culture device 105 transfer samples via a compartment transfer device 205 , and the sample processing device 104 and the sample monitoring device 106 transfer samples via a compartment transfer device 206 .

[0021] In some embodiments, the apparatus further comprises a base 103 , and the moving device comprises a moving robot arm and a track device for transferring samples within and between each device, and the track device is located on the base 103 .

[0022] In some embodiments, the mobile robotic arm includes a mobile robotic arm 402 for cell culture fluid samples, a mobile robotic arm 403 for expanded culture, a mobile robotic arm 404 for cell samples, a mobile robotic arm 405 for transfection samples, a mobile robotic arm 406 for a waste collection device, a mobile robotic arm 407 for sample transfer within the cell culture device 105, and a mobile robotic arm 408 for sample transfer within the sample monitoring device 106.

[0023] In some embodiments, the track device includes a track device 503 for cell culture fluid samples, a track device 504 for transferring cell expansion culture samples, a track device 505 for transferring cell samples, a track device 506 for monitoring sample transfer, a track device 507 for transferring a waste collection device, and track devices 501 and 502 for moving compartment transfer devices 205 and 206.

[0024] In some embodiments, the track device is, for example, a chain conveyor belt.

[0025] In some embodiments, the device further comprises a control system device for controlling the operation of each device in the device. For example, the control system device is used to control the cell culture temperature, culture time, etc.

[0026] In some embodiments, the control system device is not in communication with the devices within the apparatus.

[0027] A second aspect of the present invention provides a method for automatically expressing a recombinant protein in a cell, the method comprising the following steps:

[0028] Step 1) sterilizing the inside of the device by means of a sterilizing device 311;

[0029] Step 2) Maintain unidirectional airflow inside the equipment through air circulation devices;

[0030] Step 3) using a mobile robot and a track device to transfer the sample from the sample pretreatment device to the cell culture fluid area 5003);

[0031] Step 4) adding the sample into the cell culture medium through a sample adding device;

[0032] Step 5) using a mobile robot and a track device to transfer the sample to the compartment transfer devices 205 and 206, and transferring the sample to the cell culture device 105 through the track device for culture and cell expansion;

[0033] Step 6) transferring the cell culture fluid from the cell culture device 105 to the sample processing device 104 by moving the mechanical arm and the track device, taking a sample by the sampling device, transferring the sample to the sample monitoring device 106 by moving the mechanical arm and the track device, and using the sample monitoring device 106 to detect the cell concentration and growth status in the culture fluid;

[0034] Step 7) transferring the cell culture fluid from the cell culture device 105 to the sample processing device 104 by moving the robot arm and the track device, expanding the culture fluid by the sample adding device, and then transferring the cell culture fluid to the cell culture device 105 by moving the robot arm and the track device for further culture;

[0035] Step 8) When the cell growth reaches a predetermined value, a mobile robot is used to transfer a larger volume of cell culture fluid from the sample pretreatment device to the sample treatment device, and the mobile robot and the track device are used to transfer the cell culture fluid to the sample treatment device 104, and the sample adding device is used to mix the cell culture fluid to form a new cell culture fluid;

[0036] Step 9) adding the transfection reagent to the new cell culture solution through the sample adding device and mixing, and transferring the mixed solution to the cell culture device 105 for further culture;

[0037] Step 10) After transfection, the ambient temperature is adjusted by the temperature and humidity monitoring and control device 3012, and samples are obtained from the cell culture device 105 and transferred to the sample monitoring device 106 at regular intervals;

[0038] Step 11) the sample monitoring device 106 continues to monitor cell growth and transfection effect;

[0039] Step 12) When the protein expression reaches the expected level, the biomass is transferred from the cell culture device 105) to the sample processing device and then transferred outside the device using a mobile robot and a track device.

[0040] In some embodiments, after cell expansion is completed in step 11), the cells express the recombinant protein for 1 hour to 1 week, and the time can be 1 hour, 2 hours, 3 hours or 1 day, 2 days, 3 days, or 7 days.

[0041] In some embodiments, the device is the device as described in the first aspect of the present invention.

[0042] Cell culture equipment: The cell culture equipment is designed to provide an optimal environment for cell culture and expansion. It ensures that the cell culture is properly aerated and mixed, promoting growth and biomass production, such as CO 2 Incubators, shakers, mixers, etc.

[0043] Sample monitoring device: used to monitor cell concentration in culture to ensure optimal growth conditions and determine when biomass can be harvested. It can also detect recombinant protein expression in culture to ensure sample harvesting conditions, such as spectrophotometers and flow cytometers.

[0044] Sample pretreatment device: The sample pretreatment device is used for sample treatment before cell culture, including steps such as cell separation, purification, counting and dilution, and can transfer biological samples outside and inside the device to avoid cross contamination between the external and internal environments. It includes a cell counter: used to count the number of cells and evaluate cell activity; a centrifuge: used for cell precipitation and separation; a filter: used to remove impurities and cell debris in the sample; a purification column: used for cell purification; and a diluter: used to adjust the concentration of the cell suspension.

[0045] The cell culture fluid pretreatment device is used for sample processing of the cell culture fluid, including steps of preparing, mixing, diluting the culture fluid, etc. The cell culture fluid can be transferred outside and inside the device to avoid cross contamination between the external and internal environments.

[0046] The cell sample pretreatment device is used for processing the cell sample, including steps such as sample resuspension, mixing, and dilution. The cell sample can be transferred outside and inside the device to avoid cross contamination between the external and internal environments.

[0047] The transfection sample pretreatment device is used for processing the transfection sample, including steps such as sample resuspension, mixing, and dilution. The transfection sample can be transferred outside and inside the device to avoid cross contamination between the external and internal environments.

[0048] The waste pretreatment device is used for the treatment of waste collection devices, including steps such as cleaning, disinfection and sterilization of the device. The waste device can be transferred outside and inside the equipment to avoid cross contamination between the external and internal environments.

[0049] Compartment transfer device: A space used to transfer biological samples between different compartments inside the device to avoid cross contamination between different compartments.

[0050] Sample loading device: used to accurately add cell suspension, culture medium, reagents, etc. into culture containers. It is a device that uses experimental consumables to transfer liquids and add samples in a fixed area, such as area positioning pipette heads. Manual pipettes are suitable for small-scale operations; electronic pipettes provide more precise volume control; multichannel pipettes are suitable for high-throughput sample loading.

[0051] Consumables storage device: used to store unused experimental consumables in a fixed location to reduce the risk of cross-contamination between areas, such as gun tips.

[0052] Waste collection device: used to collect used experimental consumables. The waste collection device is located at the downwind outlet to minimize the contamination of the experimental process by waste liquid and waste, such as waste liquid barrels and waste consumables recycling barrels.

[0053] Air circulation device: Maintains environmental cleanliness and forms one-way airflow to minimize cross-contamination between different areas inside the equipment.

[0054] Sterilization device: used for environmental sterilization to ensure the sterile environment required in the process. Specific implementation methods include ultraviolet irradiation and high-temperature steam.

[0055] Temperature and humidity monitoring and control device: used to control compartment temperature to ensure cell growth and protein activity.

[0056] Mobile Robot: A robot located on a fixed track that moves biological samples between the different parts of the device. The robot is equipped with a sample gripper that safely grips and moves the sample tubes, while automating the opening and closing of the sample cover. It ensures precise manipulation and minimizes the risk of contamination.

[0057] Track device: An automated track transport system located under the robotic arm for sample movement. The system is responsible for ensuring that the sample container is smoothly transported to the next processing area after the sample is opened or closed. The track system maintains the stability of sample transportation throughout the process, improving operational efficiency and reliability.

[0058] The area positioning system is located next to the sample loading device and is used to identify the corresponding specific area below and the position of the experimental consumables in the area, so that the sample loading device can accurately locate and perform operations such as sample loading and pipetting.

[0059] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0060] The reagents and raw materials used in the present invention are commercially available.

[0061] The positive and progressive effects of the present invention are:

[0062] 1. An efficient clean system has been introduced, which can provide a stable clean environment throughout the entire production process and effectively prevent cross contamination. The clean system ensures the cleanliness of the compartments through multi-layer filtration devices, air circulation systems, and regular sterilization operations, and minimizes the risk of contamination between different process flows through the design of one-way airflow. Each production compartment operates relatively independently, which enables processes such as cell culture, protein expression, testing, and harvesting to be interconnected, and the combination of different processes is flexible and feasible, which improves production efficiency and the stability of product quality.

[0063] 2. Achieved a high degree of multifunctionality: The present invention integrates functions such as cell culture, monitoring, transfection, waste collection, gas management, sample transportation, sterilization, temperature and humidity control into a compact device, replacing the traditional complex layout that requires a large area of ​​sterile laboratories. This integration simplifies operations, improves the level of automation, greatly improves production efficiency, and reduces land occupation and resource requirements.

[0064] 3. It solves the technical problems of low efficiency, easy errors, and difficult control of production efficiency and product quality in traditional manual operations. It realizes the automation, high efficiency, controllability and large-scale production of the cell expression of recombinant proteins, and improves the production efficiency and product quality of the biopharmaceutical industry.

[0065] 3. It integrates multiple steps such as cell amplification, transfection, cell growth level detection, protein expression level detection, etc., which is convenient for operation and management.

[0066] 4. The main body of the equipment adopts a multi-compartment structure to achieve automation, integration and compact layout of each process of cell protein expression. A robotic arm and track system are used to transfer samples between different compartments, automate the process and improve production efficiency. Each compartment is relatively independent, achieving environmental sterilization and temperature control during the production process, ensuring the cleanliness and constant temperature of the production environment and improving product quality.

[0067] 5. The conditions for protein expression products can be screened by controlling the process parameters, such as temperature and time. The automation of the protein expression process can be achieved by using automated cell culture equipment.

[0068] 6. The specified volume of biomass can be generated according to user needs to meet the needs of different production scales. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] Figure 1 The following is a flow chart of the method for automatically expressing recombinant proteins in cells in the examples.

[0070] Figure 2 Schematic diagram of the structure of the device for expressing recombinant protein by cells in the embodiment.

[0071] Figure 3 Schematic diagram of the internal structure of the device for expressing recombinant protein by cells in the embodiment, parts B, D and E are top views.

[0072] Figure 4 Schematic diagram of a sample corresponding to the sample adding device in the embodiment. DETAILED DESCRIPTION

[0073] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0074] Example 1: Device for Automating Cell Expression of Recombinant Proteins

[0075] The present invention provides an automated cell expression recombinant protein device, such as Figure 2 As shown, the outer layer is a closed shell, including a fixed shell 101, an openable shell 102, a base 103 and a control panel 100. The device includes multiple independent compartments, including a sample processing device 104, a cell culture device 105, a sample monitoring device 106, compartment transfer devices 205 and 206, a moving device, a sterilizing device 311 and an air circulation device, the air circulation device includes an air inlet device 301 and an air outlet device 302, the air inlet device 301 and the air outlet device 302 can be connected to the outside of the device, the air inlet device 301 is provided with an air inlet 107, and the air outlet device 302 is provided with an air outlet 108. The air outside the device enters the air inlet device 301 through the air inlet 107, and is discharged from the device through the air outlet 108 after passing through the air outlet 302, forming a unidirectional airflow from one side to the other side in the device, and a filter component is provided in the air inlet device 301, and the filter component can be selected as a HEPA filter, and the air outside the device enters the device after passing through multiple layers of filtration.

[0076] like Figure 3 As shown, a sample pretreatment device for a sample is provided in the sample processing device 104, and the sample pretreatment device includes, from right to left, a pretreatment device 201 for a cell culture fluid, a pretreatment device 202 for a cell sample, a pretreatment device 203 for a transfected sample, and a pretreatment device 204 for a waste. The sample pretreatment device can be opened from the front exterior of the device, and from the upper portion of the device interior, and the sample can be transferred into or out of the device through the sample pretreatment device. Biological samples are transferred between devices through compartment transfer devices 205 and 206, and the compartment transfer devices can be opened at the upper portion of the compartments for sample transfer, wherein the sample processing device 104 and the cell culture device 105 transfer samples through the compartment transfer device 205, and the sample processing device 104 and the sample monitoring device 106 transfer samples through the compartment transfer device 206.

[0077] The equipment is provided with an air circulation device and a sterilization device 311 for achieving cleanliness and sterility of the internal environment of the equipment. The air circulation device is arranged in the sample processing device 104, and a one-way airflow is formed inside by the air inlet device 301 and the air outlet device 302. Specifically, the air is introduced into the air inlet 107 after passing through the external multi-layer filtration, forming a one-way airflow from one side to the other side, and is discharged from the equipment through the air outlet 108. Each functional compartment is provided with an independent sterilization device 311, which is arranged in the sample processing device 104, the cell culture device 105, the sample monitoring device 106, each sample pretreatment device, and the compartment transfer devices 205 and 206. The cell culture device 105 is also provided with a temperature and humidity monitoring and control device 3012 for maintaining a stable environment during the cell culture process.

[0078] The device is provided with a mobile device, including a mobile mechanical arm and a track device, and a waste collection device for collecting waste generated in the experiment, and the waste collection device is located in the sample processing device 104. The mobile mechanical arm is located above the device and the sample, and is used to grab the sample and transfer it to a suitable position. It includes a mobile mechanical arm 402 for cell culture fluid samples, a mobile mechanical arm 403 for expanded culture, a mobile mechanical arm 404 for cell samples, a mobile mechanical arm 405 for transfected samples, a mobile mechanical arm 406 for a waste collection device, a mobile mechanical arm 407 for sample transfer in the cell culture device 105, and a mobile mechanical arm 408 for sample transfer in the sample monitoring device 106. The track device is located on the base 103 and is used for transferring samples between the sample loading device and other areas and for moving the compartment transfer devices 205 and 206. The track device includes a track device 503 for cell culture fluid samples, a track device 504 for transferring cell expansion culture samples, a track device 505 for transferring cell samples, a track device 506 for monitoring sample transfer, a track device 507 for transferring a waste collection device, and track devices 501 and 502 for moving the compartment transfer devices 205 and 206.

[0079] Inside the sample processing device 104, there is a special consumable storage device, which is used for storage areas 603 and 604 of different experimental consumables to ensure the classification and use of experimental consumables. Area 601 in the cell culture device 105 is used to store equipment and instruments related to cell culture. Area 602 in the sample monitoring device 106 is used to store related monitoring instruments, which are used to monitor the growth and induction status of cells.

[0080] The sample processing device 104 is divided into regions as follows: Figure 4As shown, it includes different experimental consumables areas 6003 and 6004, monitoring and transfection sample loading area 5006, cell culture fluid area 5003, cell sample area 5005 and waste collection area 5007. The sample processing device 104 is provided with a sample loading device and a regional positioning system 401, and the regional positioning system 401 is equipped with a pipette head 4001. The sample loading device draws liquid from one area through the pipette head 4001 equipped with the regional positioning system 401 and adds it to the sample in another area.

[0081] A control system device is provided in the device, and the control system device is connected to the control panel 100. According to the requirements input by the user on the control panel 100, the raw materials required for protein expression are provided, including the required cell samples, cell culture well plates, culture fluids and other materials. If the consumables of the consumable storage device of the device are insufficient, a prompt is given. The control system device can control subsequent automatic transportation, detection and analysis, intelligent judgment and other operations.

[0082] Example 2: Methods and steps for expressing recombinant proteins in cells

[0083] The process of automated cell expression of recombinant proteins is as follows Figure 1 shown.

[0084] Cell expansion culture: The reagents and consumables required for the experiment are added to the experimental consumables storage areas 603 and 604 through the openable shell 102. The inside of the device is sterilized by the sterilization device. The unidirectional airflow inside the device is maintained by the air circulation device. The cell sample is placed in the pretreatment device 202, and the cell culture solution is placed in the pretreatment device 201. The device transfers the cell sample from the pretreatment device 202 to the cell sample area 5005 by moving the mechanical arm 404 and the track device 505, and transfers the cell culture solution from the pretreatment device 201 to the cell culture solution area 5003 by moving the mechanical arm 402 and the track device 503. The sample adding device obtains the experimental consumables from the area 604, and the sample is sucked from the cell sample area 5005 by the pipette head 4001 and added to the cell culture solution area 5003 to obtain the cell solution for expansion culture, and the experimental waste is transferred to the waste collection area 5007. The expanded cultured cell solution is then transferred to the compartment transfer device 205 through the track device 503 and the mobile robot 402, and then enters the cell culture device 105 through the track device 501. It is transferred to the area 601 of the cell culture device through the mobile robot 407. The temperature and humidity inside the device are controlled by the temperature and humidity monitoring and control device 3012, and the cells are expanded and cultured.

[0085] Cell concentration monitoring: The cell culture sample is transferred to the cell culture fluid area 5003. The sample loading device obtains consumables from the area 603, obtains a small amount of monitoring sample from the cell culture fluid area 5003, and transfers it to the monitoring and transfection sample loading area 5006. The monitoring sample is transferred to the compartment transfer device 206 through the track device 506 and the mobile mechanical arm 405, and then enters the sample monitoring device 106 through the track device 502. It is transferred to the area 602 in the sample monitoring device 106 through the mobile mechanical arm 408. If the cell concentration is not reached, the cell sample is transferred back to the cell culture device 105; if it meets the requirements, it enters the next stage.

[0086] Cell transfection: The cell culture sample is transferred to the sample processing device 104, and is transferred to the cell sample area 5005 by the mobile robot 402, the track device 503, the mobile robot 403, 504, 404 and 505. The new cell culture fluid is placed in the pretreatment device 201 and transferred to the cell culture fluid area 5003. The transfection reagent is placed in the pretreatment device 203 and is transferred to the monitoring and transfection sample loading area 5006 by the mobile robot 405 and the track device 506. The samples are mixed to obtain the transfection culture fluid in the cell culture fluid area 5003, and the culture fluid is transferred to the cell culture device 105 for culture.

[0087] After transfection, the ambient temperature is adjusted by the temperature and humidity monitoring and control device 3012, and the cell sample is transferred from the cell culture device 105 to the sample processing device 104 at a certain interval, and the sample is transferred to the sample monitoring device 106 to detect the cell growth OD value and the recombinant protein expression level.

[0088] When the cell growth reaches a certain value, the cells are expanded and cultured as needed. After completion, the cells are allowed to express the recombinant protein for an appropriate period of time, which can range from a few hours to a few days.

[0089] When the cell growth and recombinant protein expression reach the expected level, the biomass produced by the cell culture is transferred from the cell culture device 105 to the pretreatment device 201 using a robotic arm, and then transferred out of the device.

[0090] Although the preferred embodiments of the present invention have been described in detail, it should be understood that various modifications and changes can be made to it without departing from the spirit and scope of the present invention. Therefore, the present invention should not be limited to the specific embodiments described herein, but should be limited to the full scope of the appended claims and their equivalents.

Claims

1. A device for expressing recombinant proteins in cells, characterized in that: The device comprises a plurality of independent compartments, wherein different compartments of the device are provided with a sample processing device (104), a cell culture device (105), a sample monitoring device (106), a compartment transfer device (205 and 206) and a sterilization device (311); the cell culture device (105) is provided with a temperature and humidity monitoring and control device (3012); the sample processing device (104), the cell culture device (105), the sample monitoring device (106) and the compartment transfer devices (205 and 206) are provided with a sterilization device (311); An air circulation device is provided inside the sample processing device (104), and the air circulation device comprises an air inlet device (301) and an air outlet device (302). The portion of the air inlet device (301) connected to the device housing is provided with an air inlet (107), and the portion of the air outlet device (302) connected to the device housing is provided with an air outlet (108). Air outside the device enters the air inlet device (301) through the air inlet (107), passes through the air outlet device (302), and is discharged from the device through the air outlet (108), thereby forming a unidirectional airflow from one side to the other side inside the device.

2. The device according to claim 1, characterized in that The apparatus further comprises a mobile device; and / or, The air inlet device (301) is provided with a filter component; and / or, The compartment transfer devices (205) and (206) transfer between different compartments; and / or, The sample processing device (104) is further provided with a sample pre-processing device, a sample adding device, a waste collection device and a consumable material storage device; and / or, The device further comprises a control system device; and / or, The compartments are each provided with a sterilization device (311).

3. The device according to claim 2, characterized in that The sample pretreatment device is arranged in an independent compartment; and / or, The sample pretreatment device comprises a cell culture fluid pretreatment device (201), a cell sample pretreatment device (202), a transfection sample pretreatment device (203) and a waste pretreatment device (204); and / or, The sample processing device (104) includes different experimental consumables areas (6003) and (6004), a monitoring and transfection sample loading area (5006), a cell culture fluid area (5003), a cell sample area (5005) and a waste collection area (5007).

4. The device according to claim 2 or 3, characterized in that The consumables storage device comprises storage areas (603) and (604) for different experimental consumables, ensuring the classified placement and use of experimental consumables; the cell culture device (105) comprises an area (601) for storing equipment and instruments related to cell culture; the sample monitoring device (106) comprises an area (602) for storing monitoring instruments for monitoring cell growth and induction status; and / or, The sample processing device 104 is provided with a sample adding device and a regional positioning system (401). The regional positioning system (401) is equipped with a liquid transfer head (4001). The sample adding device draws liquid from one area through the liquid transfer head (4001) equipped with the regional positioning system (401) and adds the liquid to a sample in another area.

5. The device according to any one of claims 1 to 4, characterized in that: The sample processing device (104) and the cell culture device (105) perform sample transfer via a compartment transfer device (205), and the sample processing device (104) and the sample monitoring device (106) perform sample transfer via a compartment transfer device (206); and / or, The device further comprises a base (103), and the moving device comprises a moving mechanical arm and a track device for transferring samples within and between the devices, wherein the track device is located on the base (103).

6. The device according to claim 5, characterized in that The mobile robotic arm includes a mobile robotic arm (402) for cell culture fluid samples, a mobile robotic arm (403) for expanded culture, a mobile robotic arm (404) for cell samples, a mobile robotic arm (405) for transfected samples, a mobile robotic arm (406) for a waste collection device, a mobile robotic arm (407) for sample transfer within a cell culture device (105), and a mobile robotic arm (408) for sample transfer within a sample monitoring device (106); and / or, The track device includes a track device (503) for cell culture fluid samples, a track device (504) for transferring cell expansion culture samples, a track device (505) for transferring cell samples, a track device (506) for monitoring sample transfer, a track device (507) for transferring waste collection devices, and track devices (501) and (502) for moving compartment transfer devices (205) and (206).

7. The device according to any one of claims 1 to 6, characterized in that: The device also includes a control system device, which is used to control the operation of each device in the device; there is no communication between the control system device and the devices in the device.

8. A method for automatically expressing recombinant proteins in cells, characterized in that: The method comprises the following steps: Step 1) sterilize the inside of the equipment through a sterilization device; Step 2) Maintain unidirectional airflow inside the equipment through air circulation devices; Step 3) using a mobile robot and a track device to transfer the sample from the sample pretreatment device to the cell culture fluid area; Step 4) adding the sample into the culture medium of the cells to be cultured through a sample adding device; Step 5) using a mobile robot and a track device to transfer the sample to a compartment transfer device, and transferring the sample to a cell culture device through the track device for culture and cell expansion; Step 6) transferring the cell culture fluid from the cell culture device to the sample processing device by moving the mechanical arm and the track device, taking samples by the sampling device, transferring the taken samples to the sample monitoring device by moving the mechanical arm and the track device, and using the sample monitoring device to detect the cell concentration and growth status in the culture fluid; Step 7) transferring the cell culture fluid from the cell culture device to the sample processing device by moving the robot arm and the track device, expanding the culture fluid by the sample adding device, and then transferring the cell culture fluid to the cell culture device for further culture by moving the robot arm and the track device; Step 8) When the cell growth reaches a predetermined value, a mobile robot is used to transfer a larger volume of cell culture fluid from the sample pretreatment device to the sample treatment device, and the mobile robot and the track device are used to transfer the cell culture fluid to the sample treatment device, and the sample addition device is used to mix the cell culture fluid to form a new cell culture fluid; Step 9) adding the transfection reagent to the new cell culture solution through the sample adding device and mixing, and transferring the mixed solution to the cell culture device for further culture; Step 10) After transfection, the ambient temperature is adjusted by a temperature and humidity monitoring and control device, and samples are taken from the cell culture device at regular intervals and transferred to a sample monitoring device; Step 11) The sample monitoring device continues to monitor cell growth and transfection effect; Step 12) When protein expression reaches the expected level, a mobile robotic arm and track device are used to transfer the biomass from the cell culture device to the sample processing device and then to the outside of the device.

9. The method according to claim 8, characterized in that After cell expansion is completed in step 11), the cells express the recombinant protein for a period of 1 hour to 1 week, which may be 1 hour, 2 hours, 3 hours, or 1 day, 2 days, 3 days, or 7 days.

10. The method according to claim 8 or 9, characterized in that The device is a device as described in any one of claims 1-7.

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