Modular purification device and purification method

Abstract

The application relates to the technical field of biological medicine separation and purification, and discloses a modular purification device and a purification method.The device comprises a mounting base connected with a power supply, a plurality of module mounting seats, a data connection port and a controller, the module mounting seat is provided with an electric connection port to be connected with the controller and the power supply, the data connection port is connected with the controller and external equipment, a modular functional module is detachably connected with the module mounting seat and is provided with an electric connection head, the modular functional module comprises a pumping module, a valve group module and a sensor module, all of the three modules are provided with a pipeline quick connector, an ultrafiltration and concentration unit is provided with a pipeline quick connector, the sensor module is used for detecting the pressure of input and output ends of the ultrafiltration and concentration unit, a weighing unit is used for weighing the weight of filtered liquid, a pipeline is detachably connected with the pipeline quick connector so as to connect the modular functional module and the weighing unit to form a purification system.The modular purification device can realize quick disassembly and assembly of the purification system and has high adaptability.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical separation and purification technology, specifically to a modular purification device, and also to a purification method. Background Technology

[0002] Tangential flow filtration (TFF) technology is a key unit operation in downstream purification processes in biopharmaceuticals, widely used for the concentration, desalting, and buffer replacement of biopharmaceuticals such as monoclonal antibodies, recombinant proteins, and viral vectors. Currently, industrially applied TFF systems typically consist of basic units such as pumps, pressure sensors, membrane envelopes or hollow fiber membrane columns, and associated piping and valves.

[0003] However, existing technologies still have significant limitations in achieving high levels of automation, process robustness, and system flexibility. Firstly, at the process control level, most systems rely on manual intervention and experience-based judgment by operators. For example, maintaining a key parameter for process stability—the transmembrane pressure differential—usually requires manual reading of pressure gauges installed in a dispersed manner and coarse control through manual valve adjustment. This approach not only introduces human error, making it difficult to guarantee batch-to-batch process consistency, but also increases the workload and exposure risks for workers. Although some technologies have attempted to automatically control fluid flow rates (e.g., the scheme described in publication number CN112588116B), their control dimensions are relatively singular, failing to achieve an integrated closed-loop control strategy based on real-time feedback from multiple parameters such as the transmembrane pressure differential (TMP).

[0004] Secondly, regarding system integration and compatibility, existing devices are typically designed for specific membrane modules or fixed process paths. Their hardware interfaces and fluid paths are relatively fixed, making it difficult to quickly adapt to membrane separation devices of different brands, specifications, or applications. This is particularly inconvenient in scenarios requiring frequent process changes or R&D scale-up. Although designs using disposable components exist to reduce the risk of cross-contamination, their focus is on the physical integration of the pump and membrane capsule. Complete solutions have not yet been provided for modular expansion capabilities, plug-and-play functionality of different functional units (such as different process valve groups and sensors), and flexible reconfiguration of the overall flow path topology.

[0005] In addition, existing technical solutions typically operate the TFF system as an independent unit, and its connection and data exchange with other process units such as upstream bioreactors and downstream chromatography systems are not smooth enough, making it difficult to support the construction of a continuous and integrated bioprocess platform.

[0006] In summary, existing tangential flow purification technologies still have room for improvement in terms of fully automated closed-loop control, modular and flexible configuration, and seamless integration with upstream and downstream processes. Summary of the Invention

[0007] The purpose of this invention is to overcome the problems of low automation, poor process stability, insufficient system compatibility and flexibility, and difficulty in seamless integration with upstream and downstream processes in existing technologies, and to provide an automated tangential flow purification system and its applications. This system can achieve a high degree of automation and precise control of the entire tangential flow purification process, improve batch-to-batch consistency and product yield, and has good equipment compatibility and process scalability to meet the purification needs of different scales and diverse products from R&D to production.

[0008] To achieve the above objectives, a first aspect of the present invention provides a modular purification apparatus, comprising: The mounting base is connected to a power source and is provided with multiple module mounting bases, data connection ports, and a controller. The module mounting bases are provided with electrical connection ports and are connected to the controller and the power source via the electrical connection ports. The data connection ports are used to connect to the controller and external devices. Multiple modular functional modules are detachably connected to the module mounting base, and each modular functional module is provided with an electrical connector that connects to the electrical connection port, so as to establish power connection and communication connection through the cooperation of the electrical connection port and the electrical connector; the modular functional module includes a pumping module, a valve group module and a sensor module, and the input and output ends of the pumping module, the valve group module and the sensor module are all provided with quick-connect pipe interfaces; An ultrafiltration concentration unit, wherein both the input and output ends of the ultrafiltration concentration unit are provided with the quick-connect pipe, and the sensor module is used to detect the pressure at the input and output ends of the ultrafiltration concentration unit; A weighing unit is used to weigh the filtrate and is connected to the controller; The pipe is detachably connected to the quick-connect interface of the pipe and connects the modular functional module and the weighing unit via the pipe to form a purification system.

[0009] Optionally, the pumping module includes a circulation pump, both the input and output ends of which are provided with the pipe quick-connect interface, and the input end of the circulation pump is connected to the container of the liquid to be filtered via the pipe, so as to pump the liquid to be filtered in the container into the purification system.

[0010] Optionally, the ultrafiltration concentration unit is mounted on the mounting base. The ultrafiltration concentration unit includes a receiving tube and a hollow fiber membrane disposed within the receiving tube. The receiving tube has an inlet, a circulation outlet, and a discharge outlet. The inlet and the discharge outlet are located at opposite ends of the receiving tube, and the circulation outlet is located at the same end of the receiving tube as the inlet. The quick-connect pipe is provided at the inlet, the circulation outlet, and the discharge outlet. The inlet is connected to the output end of the circulating pump via the pipe; The discharge outlet is connected to the weighing unit via the pipe; The circulating outlet is connected to the container for the liquid to be filtered via the pipe.

[0011] Optionally, the sensor module includes a first sensor and a second sensor. The first sensor is connected to the inlet via the pipe to detect the pressure at the inlet. The second sensor is connected to the outlet via the pipe to detect the pressure at the outlet.

[0012] Optionally, the valve assembly module includes a proportional valve connected via the pipe between the output of the circulating pump and the inlet, and the controller controls the opening degree of the proportional valve in response to the detection data of the first sensor and the second sensor.

[0013] Optionally, the weighing unit includes a weighing device and a collector disposed on the weighing device, the discharge outlet is connected to the collector via the pipe, and the weighing device is used to weigh the filtrate collected in the collector.

[0014] Optionally, the pumping module further includes a displacement pump, the displacement pump having both an input and an output end equipped with the pipe quick-connect interface, the input end of the displacement pump being connected to a buffer container containing buffer solution via the pipe, and the output end of the displacement pump being connected to the input end of the circulation pump or the inlet via the pipe to pump the buffer solution in the buffer container into the purification system.

[0015] Optionally, a pressure reducing valve is also included, wherein both the input and output ends of the pressure reducing valve are provided with the pipe quick-connect interface, the input end of the pressure reducing valve is connected to the gas source via the pipe, and the output end of the pressure reducing valve is connected to the purification system via the pipe.

[0016] Optionally, it also includes a display interaction device, which is connected to the power supply and the controller to receive detection signals transmitted by the controller for display and to send the recorded operation commands to the controller.

[0017] Optionally, the valve group module further includes an on / off valve, the on / off valve having the pipeline quick-connect interface at both its input and output ends, and the on / off valve being provided at the input ends of the circulation pump and the displacement pump, as well as at the discharge outlet.

[0018] Optionally, the valve assembly module further includes a clamp valve, which includes a receiving groove suitable for accommodating the pipe, an eccentric wheel, and a drive motor. The eccentric wheel is connected to the output end of the drive motor so as to drive the eccentric wheel to rotate via the drive motor. The eccentric wheel is disposed in the receiving groove and can abut against the pipe so as to control the opening and closing of the pipe when the drive motor drives the eccentric wheel to rotate. The clamp valve is provided at the input end of the circulation pump and the displacement pump, as well as at the discharge outlet.

[0019] A second aspect of the present invention provides a purification method, implemented using the modular purification device described above, characterized in that the method includes the following steps: S1. Set process parameters: Input process parameters into the controller. The process parameters include the target value of transmembrane pressure difference (TMP), the target concentration factor, and the number of displacements. S2, Ultrafiltration Concentration: The circulation pump is started for circulation; the controller calculates the real-time transmembrane pressure difference (TMP) value in real time based on the measurement value of the sensor module, and the controller adjusts the opening of the proportional valve in response to the difference between the real-time transmembrane pressure difference (TMP) value and the target value of the transmembrane pressure difference (TMP) value, so as to maintain the difference between the real-time transmembrane pressure difference (TMP) value and the target value of the transmembrane pressure difference (TMP) value within a preset range; at the same time, the weight of the filtrate is measured in real time via the weighing unit. S3. Perform buffer replacement: When the target concentration factor is reached based on the weight of the filtrate, the purification system is switched to replacement mode by the controller, and the buffer addition and concentration operations are performed cyclically according to the number of replacements. S4. Perform material recovery: After all replacement cycles are completed, gas is introduced into the purification system through the pressure reducing valve to recover the residual material in the pipeline of the purification system by purging. S5. Post-processing: The purification system is controlled by the controller to perform cleaning and evacuation operations in sequence, and a pressure maintenance test is performed to complete the system integrity verification.

[0020] Optionally, the real-time transmembrane pressure difference (TMP) value is: TMP = (PT2 + PT1) / 2; PT1 and PT2 are the detection values ​​of the first sensor and the second sensor, respectively, and the target value of the transmembrane pressure difference TMP is set to 30-50 kPa.

[0021] Optionally, the target concentration factor is 10-50 times.

[0022] Optionally, the number of permutations is 2-10.

[0023] Through the above technical solution, the modular purification device provided by the first aspect of the present invention, by setting multiple module mounting bases, data connection ports and controllers on the mounting base, enables different functional modules such as pumping modules, valve group modules and sensor modules to be quickly and detachably connected to the module mounting bases. Data transmission between the sensor module and the controller, as well as the controller's control connection to the pumping module and valve group module, can be achieved through the cooperation of electrical connectors connected to the electrical connection ports. Furthermore, the input and output ends of the pumping module, valve group module, sensor module and ultrafiltration concentration unit are all equipped with quick-connect pipe interfaces, thereby allowing for the rapid selection of corresponding functional modules according to actual needs and the rapid construction of different purification systems through the cooperation of pipes and quick-connect pipe interfaces. This makes the modular purification device more applicable, the speed of purification system construction faster, and the constructed purification system capable of automatic control. In addition, the data connection ports facilitate data interconnection between the modular purification device and external devices.

[0024] The purification method provided in the second aspect of the present invention can realize the automated control of the purification process, and can automatically perform post-purification cleaning, verification and recording, with a high degree of automation.

[0025] Other advantages of the present invention and the technical effects of preferred embodiments will be further described in the following detailed description. Attached Figure Description

[0026] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings: Figure 1 This is a front view of the modular purification device in this invention; Figure 2 This is a schematic diagram of the purification system in this invention; Figure 3 This is a left view of the modular purification device in this invention; Figure 4 This is a right view of the modular purification device in this invention; Figure 5 This is a rear view of the modular purification device in this invention; Figure 6 This is a partial cross-sectional schematic diagram of the clamp valve in the modular purification device of this invention; Figure 7This is a right view of the modular purification device in this invention.

[0027] Explanation of reference numerals in the attached figures 1. Mounting substrate; 2. Ultrafiltration concentration unit; 3. Weighing unit; 4. Circulation pump; 5. First sensor; 6. Second sensor; 7. Proportional valve; 8. Displacement pump; 9. On / off valve; 10. Pressure reducing valve; 11. Pinch valve; 111. Receiving tank; 112. Eccentric wheel; 113. Drive motor; 12. Display and interactive device. Detailed Implementation

[0028] The specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are for illustration and explanation only and are not intended to limit the present invention.

[0029] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "connection" and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to abutment; they can refer to the internal communication of two elements or the interaction between two elements. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0030] The first aspect of this invention provides a modular purification device, see [link to previous document]. Figures 1 to 4 It includes a mounting base 1, which is connected to a power source. The mounting base 1 is provided with multiple module mounting seats, data connection ports and a controller. The module mounting seats are provided with electrical connection ports and are connected to the controller and the power source through the electrical connection ports. The data connection ports are used to connect to the controller and external devices. The mounting base 1 can be configured as a hexahedral box structure and module mounting seats can be provided on any of its surfaces except the bottom surface. Multiple modular functional modules are detachably connected to the module mounting base. Each modular functional module is equipped with an electrical connector that connects to an electrical connection port for power and communication connections. The electrical connection port uses existing technology and will not be described in detail here. The modular functional modules include a pumping module, a valve group module, and a sensor module. The input and output ends of the pumping module, valve group module, and sensor module are equipped with quick-connect pipe interfaces. The quick-connect pipe interfaces use existing technology and can achieve quick and detachable connection with pipes. Ultrafiltration concentration unit 2 has quick-connect pipes at both its input and output ends, and a sensor module is used to detect the pressure at both the input and output ends of ultrafiltration concentration unit 2. Weighing unit 3 is used to weigh the filtrate and is connected to the controller; The pipe (not shown in the figure) is detachably connected to the pipe quick-connect interface and is connected to the modular functional module and weighing unit 3 via the pipe to form a purification system.

[0031] Based on the above structure, the modular purification device provided by this invention, by setting multiple module mounting bases, data connection ports, and controllers on the mounting base 1, allows different functional modules such as pumping modules, valve group modules, and sensor modules to be quickly and detachably connected to the module mounting bases. Data transmission between the sensor module and the controller, as well as the controller's control connection to the pumping module and valve group module, can be achieved through the cooperation of electrical connectors connected via electrical connection ports. Furthermore, the input and output ends of the pumping module, valve group module, sensor module, and ultrafiltration concentration unit 2 are all equipped with quick-connect pipe interfaces, allowing for the rapid selection of corresponding functional modules according to actual needs and the use of pipes and quick-connect pipe interfaces to quickly build different purification systems. This makes the modular purification device more adaptable, the purification system faster to build, and the built purification system capable of automatic control. In addition, the data connection ports facilitate data interconnection between the modular purification device and external devices.

[0032] Furthermore, the pumping module includes a circulation pump 4, which has quick-connect pipe interfaces at both its input and output ends. The input end of the circulation pump 4 is connected to the container of the liquid to be filtered via a pipe, so as to pump the liquid to be filtered in the container into the purification system for filtration.

[0033] Furthermore, the ultrafiltration concentration unit 2 is detachably mounted on the mounting base 1, allowing for the replacement of different models of ultrafiltration concentration units 2 according to purification requirements. The ultrafiltration concentration unit 2 includes a receiving tube and a hollow fiber membrane column disposed within the receiving tube. The receiving tube has an inlet, a circulation outlet, and a discharge outlet. The inlet and discharge outlet are located at opposite ends of the receiving tube, while the circulation outlet is located at the same end as the inlet. Quick-connect pipe interfaces are provided at the inlet, circulation outlet, and discharge outlet. Specifically, the length of the receiving tube can be arranged vertically, and the inlet and circulation outlet... The inlet is located at the lower end of the receiving tube, and the outlet is located at the upper end of the receiving tube to ensure that the liquid to be filtered can move from bottom to top and be fully filtered by the hollow fiber membrane, thereby improving the filtration effect. The inlet is connected to the output end of the circulating pump 4 via a pipe. The outlet is connected to the weighing unit 3 via a pipe. The circulating outlet is connected to the container of the liquid to be filtered via a pipe. It is understood that a back pressure valve can be integrated at the circulating outlet. The back pressure valve adopts existing technology to ensure that the liquid to be filtered does not flow directly back to the container of the liquid to be filtered from the circulating outlet after entering the receiving tube, thereby ensuring effective filtration.

[0034] Furthermore, the sensor module includes a first sensor 5 and a second sensor 6. The first sensor 5 is connected to the inlet via a pipe to detect the pressure at the inlet; the second sensor 6 is connected to the outlet via a pipe to detect the pressure at the outlet. Thus, the real-time transmembrane pressure difference TMP value can be determined by the pressure detected by the first sensor 5 and the second sensor 6. The real-time transmembrane pressure difference TMP value is: TMP=(PT2+PT1) / 2, where PT1 and PT2 are the detection values ​​of the first sensor 5 and the second sensor 6, respectively.

[0035] In addition, the valve assembly module includes a proportional valve 7, which is connected between the output end and the inlet of the circulating pump 4 via a pipeline. The controller controls the opening degree of the proportional valve 7 in response to the detection data of the first sensor 5 and the second sensor 6. Specifically, when the real-time transmembrane pressure difference TMP value is greater than the target value of the transmembrane pressure difference TMP, the opening degree of the proportional valve 7 is reduced to prevent insufficient filtration; when the real-time transmembrane pressure difference TMP value is less than the target value of the transmembrane pressure difference TMP, the opening degree of the proportional valve 7 is increased to improve the filtration efficiency.

[0036] Furthermore, the weighing unit 3 includes a weighing device and a collector mounted on the weighing device. The discharge port is connected to the collector via a pipe. The weighing device is used to weigh the weight of the filtrate collected in the collector, thereby enabling the calculation of whether the target concentration factor has been reached based on the measured weight of the filtrate. The target concentration factor = W0 / (W0-W1-Wd), where W0 is the initial weight of the filtrate, Wd is the mass of the filtrate stored in the dead volume of the purification system (including pipes, containment tubes, and modular functional modules), and W1 is the weight of the filtrate.

[0037] Furthermore, the pumping module also includes a displacement pump 8. Both the input and output ends of the displacement pump 8 are equipped with quick-connect pipe interfaces. The input end of the displacement pump 8 is connected to the buffer container containing the buffer solution via a pipe, and the output end of the displacement pump 8 is connected to the input end or inlet of the circulation pump 4 via a pipe to pump the buffer solution in the buffer container into the purification system. When the output end of the displacement pump 8 is connected to the input end or inlet of the circulation pump 4 via a pipe, an existing T-connector can be used to achieve the pipe connection. The displacement pump 8 is set to be used only to pump the buffer solution into the purification system. After the buffer solution is pumped into the purification system, it will be pumped by the circulation pump 4 to circulate in the purification system to achieve further filtration and purification, thereby preventing contamination of the buffer solution in the buffer container.

[0038] Furthermore, the modular purification device also includes a pressure reducing valve 10. Both the input and output ends of the pressure reducing valve 10 are equipped with quick-connect pipe interfaces. The input end of the pressure reducing valve 10 is connected to a gas source via a pipe, and the output end of the pressure reducing valve 10 is connected to the purification system via a pipe. This allows gas to be introduced into the purification system to purge it, thereby recovering residual materials in the purification system. It can also be used to dry the purification system after cleaning.

[0039] Furthermore, such as Figure 1 As shown, it also includes a display interaction device 12, which is connected to the power supply and the controller to receive and display the detection signals transmitted by the controller, and to send the input operation commands to the controller. The display interaction device 12 can be a touch screen, which can display the detection values ​​of the first sensor 5 and the second sensor 6, as well as the real-time transmembrane pressure difference (TMP) value and the target value of the transmembrane pressure difference (TMP). It can also set the target value of the transmembrane pressure difference (TMP), the target concentration factor, and the number of replacements, etc., through the touch screen, so that the controller can control the purification system in real time based on the above parameters.

[0040] Furthermore, the valve assembly module also includes an on / off valve 9. Both the input and output ends of the on / off valve 9 are equipped with quick-connect pipe interfaces for easy connection to the purification system's pipeline. On / off valves 9 are also located at the input and discharge ports of the circulation pump 4 and the displacement pump 8. This allows the on / off valves 9 at the input and discharge ports of the displacement pump 8 to be initially adjusted to the closed state, while keeping the on / off valve 9 at the input of the circulation pump 4 in the open state. This enables the circulation pump 4 to pump liquid into the purification system's pipeline until the pressure in the purification system reaches a preset value. Afterward, the on / off valve 9 at the input of the circulation pump 4 is then adjusted to the closed state. The system is kept closed, and all on / off valves 9 are kept closed for a period of time. The pressure changes in the purification system are recorded to confirm the integrity and airtightness of the purification system. Specifically, the pressure changes in the purification system can be monitored by the first sensor 5. When the pressure in the purification system reaches the preset value of 20-50 kPa, the on / off valves 9 at the input end of the circulation pump 4 can be adjusted to the closed state and kept closed for 1-5 minutes. If the pressure drop in the purification system is within the range of 0-20 kPa, it is determined that the integrity and airtightness of the purification system are within a reasonable range.

[0041] Furthermore, the valve assembly module also includes a pinch valve 11. The pinch valve 11 includes a receiving groove 111 suitable for accommodating the pipeline, an eccentric wheel 112, and a drive motor 113. The eccentric wheel 112 is connected to the output end of the drive motor 113 so that the drive motor 113 drives the eccentric wheel 112 to rotate. The eccentric wheel 112 is located in the receiving groove 111 and can abut against the pipeline so as to control the opening and closing of the pipeline when the drive motor 113 drives the eccentric wheel 112 to rotate. The input end and the discharge outlet of the circulation pump 4 and the displacement pump 8 are both equipped with pinch valves 11, so that the pinch valves 11 at the input end and the discharge outlet of the displacement pump 8 can be adjusted to the closed state first, while keeping the pinch valve 11 at the input end of the circulation pump 4 in the open state, so that the pure liquid can be pumped through the circulation pump 4. Liquid is pumped into the pipeline of the purification system until the pressure in the purification system reaches the preset value. Then, the clamp valve 11 at the input end of the circulation pump 4 is adjusted to the closed state, and each clamp valve 11 is kept closed for a period of time. The pressure change in the purification system is recorded to confirm the integrity and airtightness of the purification system. Specifically, the pressure change in the purification system can be monitored by the first sensor 5. When the pressure in the purification system reaches the preset value of 20-50 kPa, the clamp valve 11 at the input end of the circulation pump 4 is adjusted to the closed state, and each clamp valve 11 is kept closed for 1-5 minutes. If the pressure drop in the purification system is within the range of 0-20 kPa, it is determined that the integrity and airtightness of the purification system are within a reasonable range.

[0042] A second aspect of the present invention provides a purification method, implemented using the modular purification device described above, the method comprising the following steps: S1. Set process parameters: Enter process parameters into the controller. Process parameters include the target value of transmembrane pressure difference (TMP), target concentration factor, and number of displacements. S2, Ultrafiltration Concentration: Start circulation pump 4 for circulation; the controller calculates the real-time transmembrane pressure difference TMP value in real time based on the measurement value of the sensor module, and the controller adjusts the opening of proportional valve 7 in response to the difference between the real-time transmembrane pressure difference TMP and the target value of transmembrane pressure difference TMP, so as to maintain the difference between the real-time transmembrane pressure difference TMP value and the target value of transmembrane pressure difference TMP within a preset range; at the same time, the weight of the filtrate is measured in real time via weighing unit 3. S3. Perform buffer replacement: When the target concentration factor is reached based on the weight of the filtrate, the purification system is switched to replacement mode by the controller, and the buffer addition and concentration operations are performed cyclically according to the number of replacements. S4. Perform material recovery: After all replacement cycles are completed, gas is introduced into the purification system through pressure reducing valve 10 to recover the residual material in the pipeline of the purification system by purging. S5. Post-processing: The purification system is controlled by the controller to perform cleaning and evacuation operations in sequence, and a pressure maintenance test is performed to complete the system integrity verification.

[0043] Furthermore, the target value for the transmembrane pressure difference (TMP) is: TMP = (PT2 + PT1) / 2; PT1 and PT2 are the detection values ​​of the first sensor 5 and the second sensor 6, respectively, and the target value of the transmembrane pressure difference TMP is set to 30-50 kPa.

[0044] Furthermore, the target concentration factor is 10-50 times.

[0045] Furthermore, the number of permutations is 2-10.

[0046] The modular purification apparatus provided in the first aspect of this application can be used for filtering, concentrating, and purifying lentiviruses or proteins. The lentivirus is selected from at least one of recombinant expression lentiviruses, packaging-defective lentiviruses, pseudotyped lentiviruses, targeted lentiviruses, high-titer lentiviruses, fluorescently labeled lentiviruses, and therapeutic lentiviruses. The protein is a monoclonal antibody. When used for lentivirus filtration and concentration, the molecular weight cutoff of the suitable hollow fiber membrane is 300-750 kD, and the target transmembrane pressure difference (TMP) is set to 35-45 kPa. When used for monoclonal antibodies, the molecular weight cutoff of the suitable hollow fiber membrane is 30-100 kD, and the target transmembrane pressure difference (TMP) is set to 35-45 kPa.

[0047] The following sections use the purification of proteins and lentiviruses as examples to illustrate the operation of the modular purification device described in this application: First, a modular purification device is constructed. Corresponding modular functional modules and ultrafiltration concentration units 2 are installed on the modular mounting base of the mounting substrate 1. The modular functional modules include: a circulation pump 4, a first sensor 5, a second sensor 6, a proportional valve 7, a displacement pump 8, and a clamp valve 11. The input end of the circulation pump 4 is connected to the container of the liquid to be filtered via a pipe. The receiving tube of the ultrafiltration concentration unit 2 has an inlet, a circulation outlet, and a discharge outlet. The length of the receiving tube is arranged vertically. The inlet and circulation outlet are located at the lower end of the receiving tube, and the discharge outlet is located at the upper end. The inlet is connected to the output end of the circulation pump 4 via a pipe, and the discharge outlet is connected to the weighing unit 3 via a pipe. The circulation outlet is connected to the container of the liquid to be filtered via a pipe. A back pressure valve is integrated at the circulation outlet. The first sensor 5 is connected to the inlet via a pipe to detect the pressure at the inlet. The second sensor 6 is connected to the inlet via a pipe... A proportional valve 7 is connected to the discharge port to detect the pressure at the discharge port. A proportional valve 7 is connected via a pipe between the output and inlet of the circulating pump 4. The controller controls the opening of the proportional valve 7 in response to the detection data from the first sensor 5 and the second sensor 6. The input of the displacement pump 8 is connected via a pipe to a buffer container containing the buffer solution. The output of the displacement pump 8 is connected via a pipe to either the input or inlet of the circulating pump 4. When the output of the displacement pump 8 is connected via a pipe to either the input or inlet of the circulating pump 4, an existing tee connector can be used for the pipe connection. The input of the pressure reducing valve 10 is connected via a pipe to a gas source. The output of the pressure reducing valve 10 is connected via a pipe to the purification system, specifically, it can be connected to the discharge port. The discharge port is generally located at a high position in the purification system, and connecting it to the discharge port facilitates the rapid blowing out of residual materials. Pinch valves 11 are installed at the input and discharge ports of both the circulating pump 4 and the displacement pump 8. This completes the construction of the purification device.

[0048] Protein purification: The stock solution of CHO-K1 cells was harvested after culturing in a bioreactor using the Fed-Batch process for 14 days. After centrifugation and clarification, 2000 mL of the supernatant was collected. This supernatant was used as the filtrate. The initial protein concentration was determined to be 1.65 g / L by high performance liquid chromatography (HPLC).

[0049] The modular purification device is installed and constructed according to the above installation method to form a purification system. The hollow fiber column membrane has a molecular weight cutoff of 30kD. The purification system is then cleaned. Specifically, the purification system can be cleaned by pumping purified water into the purification system using the circulation pump 4. This is existing technology and will not be described in detail here. Then, the circulation pump 4 and the displacement pump 8 are calibrated using existing technology to complete the preparation work.

[0050] Process parameters were input on the interactive display device 12. The target transmembrane pressure difference (TMP) was set to 40 ± 1 kPa, the target concentration factor to 40 times, and the number of replacements to 5. The purification system then entered the ultrafiltration concentration step, in which 50 mL of concentrate and 1960 mL of filtrate waste liquid were obtained. Next, a buffer replacement step was performed for 5 replacement washes. Each replacement involved adding 50 mL of buffer solution (PBS, phosphate buffer saline) to concentrate 100 mL of the mixture by ultrafiltration, repeating this process 5 times. Following this, a material recovery step was performed to recover the liquid from the system tubing, ultimately yielding 50 mL of product and 250 mL of filtrate waste liquid. Finally, post-treatment was performed to clean and regenerate the hollow fiber membrane: first, it was rinsed with purified water for 10 min, then rinsed with 0.5 mol / L NaOH solution for 10 min, and finally soaked in 0.1 mol / L NaOH solution.

[0051] Purification of lentiviruses: Take 200 mL of HEK293T packaged cell culture supernatant, which is the filtrate to be filtered. The initial infection titer was detected by flow cytometry as 5.5E+07 TU / mL.

[0052] The modular purification device is installed and constructed according to the above installation method to form a purification system. The hollow fiber column membrane has a molecular weight cutoff of 500kD. The purification system is then cleaned. Specifically, the system can be cleaned by pumping purified water into the purification system using the circulation pump 4. This is existing technology and will not be described in detail here. Then, the circulation pump 4 and the displacement pump 8 are calibrated using existing technology to complete the preparation work.

[0053] Process parameters were input on the interactive display device 12. The target transmembrane pressure difference (TMP) was set to 40 ± 1 kPa, the target concentration factor to be 10 times, and the number of replacements to be 5. The purification system then entered the ultrafiltration concentration step, in which 20 mL of concentrate and 180 mL of filtrate were obtained. Next, a buffer replacement step was performed for 5 replacement washes. Each replacement involved adding 20 mL of buffer solution (PBS, phosphate buffer saline) to concentrate the 40 mL mixture by ultrafiltration, repeating this process 5 times. Following this, a material recovery step was performed to recover the liquid from the system tubing, ultimately yielding 20 mL of product and 100 mL of filtrate. Finally, post-processing was performed to clean and regenerate the hollow fiber membrane: first, it was rinsed with purified water for 10 min, then rinsed with 0.5 mol / L NaOH solution for 10 min, and finally soaked in 0.1 mol / L NaOH solution.

[0054] Based on the above design, the modular purification device provided by the first aspect of the present invention, by setting multiple module mounting bases, data connection ports and controllers on the mounting base 1, enables different functional modules such as pumping modules, valve group modules and sensor modules to be quickly and detachably connected to the module mounting bases. Data transmission between the sensor module and the controller can be achieved through the cooperation of electrical connectors connected by the electrical connection ports, as well as the controller's control connection to the pumping module and valve group modules. Furthermore, the input and output ends of the pumping module, valve group module, sensor module and ultrafiltration concentration unit 2 are all equipped with quick-connect pipe interfaces, thereby allowing for the rapid selection of corresponding functional modules according to actual needs and the rapid construction of different purification systems through the cooperation of pipes and quick-connect pipe interfaces. This makes the modular purification device more applicable, the speed of purification system construction faster, and the constructed purification system capable of automatic control. In addition, the data connection ports also facilitate data interconnection between the modular purification device and external devices.

[0055] The purification method provided in the second aspect of the present invention can realize the automated control of the purification process, and can automatically perform post-purification cleaning, verification and recording, with a high degree of automation.

[0056] It should also be noted that the various specific technical features described in the above embodiments can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, the present invention will not describe the various possible combinations separately. Furthermore, various different embodiments of the present invention can also be arbitrarily combined, as long as they do not violate the spirit of the present invention, and should also be regarded as the content disclosed by the present invention.

Claims

1. A modular purification device, characterized in that, include: Mounting base (1), which is connected to a power source, and the mounting base (1) is provided with multiple module mounting seats, data connection ports and a controller. The module mounting seats are provided with electrical connection ports and are connected to the controller and the power source via the electrical connection ports. The data connection ports are used to connect to the controller and external devices. Multiple modular functional modules are detachably connected to the module mounting base, and each modular functional module is provided with an electrical connector that connects to the electrical connection port, so as to establish power connection and communication connection through the cooperation of the electrical connection port and the electrical connector; the modular functional module includes a pumping module, a valve group module and a sensor module, and the input and output ends of the pumping module, the valve group module and the sensor module are all provided with quick-connect pipe interfaces; The ultrafiltration concentration unit (2) is provided with the pipe quick-connect interface at both the input and output ends of the ultrafiltration concentration unit, and the sensor module is used to detect the pressure at the input and output ends of the ultrafiltration concentration unit. Weighing unit (3) is used to weigh the filtrate and is connected to the controller; The pipe is detachably connected to the quick-connect interface of the pipe and connects the modular functional module and the weighing unit (3) via the pipe to form a purification system.

2. The modular purification device according to claim 1, characterized in that, The pumping module includes a circulation pump (4), both the input and output ends of which are provided with the pipe quick-connect interface. The input end of the circulation pump (4) is connected to the container to be filtered via the pipe to pump the filtrate in the container into the purification system.

3. The modular purification device according to claim 2, characterized in that, The ultrafiltration concentration unit (2) is mounted on the mounting base (1). The ultrafiltration concentration unit (2) includes a receiving tube and a hollow fiber membrane disposed in the receiving tube. The receiving tube has an inlet, a circulation outlet and a discharge outlet. The inlet and the discharge outlet are respectively located at the two ends of the receiving tube. The circulation outlet and the inlet are located at the same end of the receiving tube. The quick-connect pipe is provided at the inlet, the circulation outlet and the discharge outlet. The inlet is connected to the output end of the circulating pump (4) via the pipe; The discharge outlet is connected to the weighing unit (3) via the pipe; The circulating outlet is connected to the container for the liquid to be filtered via the pipe.

4. The modular purification device according to claim 3, characterized in that, The sensor module includes a first sensor (5) and a second sensor (6). The first sensor (5) is connected to the inlet via the pipe to detect the pressure at the inlet. The second sensor (6) is connected to the outlet via the pipe to detect the pressure at the outlet.

5. The modular purification device according to claim 4, characterized in that, The valve module includes a proportional valve (7), which is connected via the pipe between the output end of the circulating pump (4) and the inlet. The controller controls the opening degree of the proportional valve (7) in response to the detection data of the first sensor (5) and the second sensor (6).

6. The modular purification apparatus according to claim 5, characterized in that, The weighing unit (3) includes a weighing device and a collector disposed on the weighing device. The discharge outlet is connected to the collector via the pipe. The weighing device is used to weigh the filtrate collected in the collector.

7. The modular purification apparatus according to claim 6, characterized in that, The pumping module also includes a displacement pump (8), the input and output ends of which are provided with the pipe quick-connect interface. The input end of the displacement pump (8) is connected to the buffer container containing the buffer solution via the pipe, and the output end of the displacement pump (8) is connected to the input end of the circulation pump (4) or the inlet via the pipe to pump the buffer solution in the buffer container into the purification system.

8. The modular purification apparatus according to claim 7, characterized in that, It also includes a pressure reducing valve (10), the input and output ends of which are provided with the pipeline quick-connect interface. The input end of the pressure reducing valve (10) is connected to the gas source via the pipeline, and the output end of the pressure reducing valve (10) is connected to the purification system via the pipeline.

9. The modular purification apparatus according to claim 8, characterized in that, It also includes a display interaction device (12), which is connected to the power supply and the controller to receive detection signals transmitted by the controller for display and to transmit recorded operation instructions to the controller.

10. The modular purification apparatus according to claim 9, characterized in that, The valve group module also includes a shut-off valve (9), the input and output ends of which are provided with the pipeline quick-connect interface. The shut-off valve (9) is provided at the input end of the circulation pump (4) and the displacement pump (8) as well as at the discharge outlet.

11. The modular purification apparatus according to claim 9, characterized in that, The valve assembly module also includes a clamp valve (11), which includes a receiving groove (111) suitable for accommodating the pipe, an eccentric wheel (112), and a drive motor (113). The eccentric wheel (112) is connected to the output end of the drive motor (113) so as to drive the eccentric wheel (112) to rotate via the drive motor (113). The eccentric wheel (112) is located in the receiving groove (111) and can abut against the pipe so as to control the opening and closing of the pipe when the drive motor (113) drives the eccentric wheel (112) to rotate. The clamp valve (11) is provided at the input end of the circulation pump (4) and the displacement pump (8) as well as at the discharge outlet.

12. A purification method, implemented using the modular purification apparatus of claim 9 or 10, characterized in that, The method includes the following steps: S1. Set process parameters: Input process parameters into the controller. The process parameters include the target value of transmembrane pressure difference (TMP), the target concentration factor, and the number of displacements. S2, Ultrafiltration Concentration: Start the circulation pump (4) for circulation; the controller calculates the real-time transmembrane pressure difference TMP value in real time based on the measurement value of the sensor module, and the controller adjusts the opening of the proportional valve (7) in response to the difference between the real-time transmembrane pressure difference TMP and the target value of the transmembrane pressure difference TMP, so as to maintain the difference between the real-time transmembrane pressure difference TMP and the target value of the transmembrane pressure difference TMP within a preset range; at the same time, the weight of the filtrate is measured in real time via the weighing unit (3); S3. Perform buffer replacement: When the target concentration factor is reached based on the weight of the filtrate, the purification system is switched to replacement mode by the controller, and the buffer addition and concentration operations are performed cyclically according to the number of replacements. S4. Perform material recovery: After all replacement cycles are completed, gas is introduced into the purification system through the pressure reducing valve (10) to recover the residual material in the pipeline of the purification system by purging. S5. Post-processing: The purification system is controlled by the controller to perform cleaning and evacuation operations in sequence, and a pressure maintenance test is performed to complete the system integrity verification.

13. The method according to claim 12, characterized in that, The real-time transmembrane pressure difference (TMP) value is: TMP = (PT2 + PT1) / 2; PT1 and PT2 are the detection values ​​of the first sensor (5) and the second sensor (6), respectively, and the target value of the transmembrane pressure difference TMP is set to 30-50 kPa.

14. The method according to claim 12, characterized in that, The target concentration factor is 10-50 times.

15. The method according to claim 12, characterized in that, The number of replacements is 2-10.

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