Efficient intelligent packaging and purifying system for oncolytic viruses

Abstract

The invention provides an efficient and intelligent oncolytic virus packaging and purifying system which comprises an intelligent virus packaging module, a totally-closed primary harvesting module, a self-adaptive chromatographic purification module and a central intelligent control system.The efficient and intelligent oncolytic virus packaging and purifying system designs a totally-closed integrated flow path, and the packaging module, the harvesting module and the purifying module are connected in series through sterile pipelines; aI image recognition is fused with UV-Vis and DLS online detection, intelligent judgment of the harvesting opportunity and purified components is achieved, and pollution caused by manual intervention is completely eradicated; manual judgment is replaced by AI detection, so that the accuracy of harvesting and purifying is greatly improved; the method has the advantages that target viruses are accurately captured through self-adaptive chromatography, the purity and the recovery rate are obviously improved, parameters are continuously optimized through machine learning, the batch difference is reduced, and a reliable scheme is provided for large-scale production of clinical oncolytic viruses.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, specifically to an efficient and intelligent packaging and purification system for oncolytic viruses. Background Technology

[0002] Oncolytic viruses, with their unique mechanism of targeting and lysing tumor cells and activating systemic anti-tumor immune responses, have become one of the core research directions in the field of tumor immunotherapy. Their clinical translation relies on stable, large-scale production processes. A typical oncolytic virus production process includes key steps such as packaging cell culture, viral plasmid transfection, viral packaging and amplification, cell lysis and harvesting, clarification of crude extract, and fine purification. However, clinical applications have extremely high requirements for product sterility (no exogenous microbial contamination), purity (residual protein / nucleic acid ≤0.1%), and batch consistency (titer variation coefficient ≤5%). Traditional general-purpose bioreactors and separation equipment are no longer suitable for their specialized and refined production needs, and there is an urgent need for targeted, integrated intelligent equipment to overcome technical bottlenecks.

[0003] Existing oncolytic virus production technologies suffer from multiple defects, the root causes of which are directly related to process design and equipment configuration: First, the fragmented process leads to significant risks of contamination and inactivation. In current production, virus packaging (bioreactors), harvesting (centrifugation equipment), and purification (chromatographic systems) are mostly handled by independent equipment, and material transfer relies on manual operation—even with aseptic connecting pipes, connections still need to be completed in a laminar flow hood, inevitably posing a risk of environmental exposure; simultaneously, the virus remains in a non-optimal temperature range (deviating from 37℃±1℃) for extended periods during transfer, easily leading to viral particle aggregation and inactivation. Current solutions involve shortening transfer time, but cannot fundamentally eliminate the "breakpoint" problem between processes; Second, the identification of critical nodes and purification control rely on manual labor and fixed procedures. Harvesting requires manual sampling and microscopic observation of cytopathic effects (CPE), with errors in CPE assessment by different operators reaching 15%-20%. The purification stage uses preset elution flow rates and buffer ratios, making it impossible to adjust strategies based on batch variations in virus concentration and contaminating protein content in the crude extract. While current attempts attempt to modify parameters after offline crude extract quality testing are time-consuming (2-4 hours) and prone to viral degradation, the system lacks collaborative optimization capabilities. Each device is equipped with an independent controller, requiring manual adjustment of parameters such as transfection voltage, dissolved oxygen, and chromatography flow rate based on experience, making it impossible to dynamically optimize downstream processes based on upstream cell status. Furthermore, there is no automatic accumulation and analysis of production data, necessitating manual summarization for process improvement, resulting in low efficiency. Summary of the Invention

[0004] (a) Technical problems to be solved

[0005] To address the shortcomings of existing technologies, this invention provides an efficient and intelligent packaging and purification system for oncolytic viruses, which solves the problems of fragmented process steps leading to significant risks of contamination and inactivation, reliance on manual labor and fixed procedures for key node judgment and purification control, and lack of collaborative optimization capabilities in existing technologies.

[0006] (II) Technical Solution

[0007] To achieve the above objectives, the present invention provides the following technical solution: an efficient and intelligent packaging and purification system for oncolytic viruses, comprising an intelligent virus packaging module, a fully enclosed primary harvesting module, an adaptive chromatography purification module, and a central intelligent control system;

[0008] The intelligent virus packaging module is used to achieve efficient transfection of plasmids into packaging cells, virus packaging culture, and to determine the optimal time to harvest the virus by real-time monitoring of cell status.

[0009] The fully enclosed primary harvest module is connected to the intelligent virus packaging module through sterile tubing and is used to receive virus culture medium, complete cell lysis, virus release and preliminary clarification of culture medium in a closed environment;

[0010] The adaptive chromatography purification module is connected to the fully enclosed primary harvest module through sterile tubing. It is used to perform multi-step fine purification of the initially clarified crude virus extract and to accurately collect the target virus fraction based on real-time detection data.

[0011] The central intelligent control system is connected to the intelligent virus packaging module, the fully enclosed primary harvesting module, and the adaptive chromatography purification module via wired or wireless communication, respectively. It is used to receive real-time operating data from each module and dynamically coordinate and optimize the process parameters of each module through a preset algorithm model.

[0012] The intelligent virus packaging module, the fully enclosed primary harvesting module, and the adaptive chromatography purification module are connected in series with a sanitary diaphragm valve via a sterile quick-connect connector, forming a continuous and closed integrated production flow path.

[0013] Preferably, the intelligent virus packaging module includes:

[0014] The bioreactor is equipped with online sensors for temperature, pH, and dissolved oxygen concentration.

[0015] An online microscopic imaging system integrated on top of the bioreactor includes an upright microscope module with 20×-40× objectives, a CMOS image sensor, and an AI image recognition unit.

[0016] A transfection unit connected to the bioreactor via sterile tubing, wherein the transfection unit is a microfluidic electrotransfection unit or a high-throughput nanomaterial transfection unit;

[0017] The AI ​​image recognition unit is used to analyze cell density, fusion degree and lesion effect in real time, and transmits the analysis results to the central intelligent control system.

[0018] Preferably, the fully enclosed primary harvesting module includes:

[0019] A sterile harvesting tank is connected to the bioreactor of the intelligent virus packaging module via sterile piping.

[0020] A processing unit integrated inside the aseptic harvesting tank, wherein the processing unit is an ultrasonic crushing unit or a tangential flow filtration unit;

[0021] The power of the ultrasonic disruption unit is adjustable from 50 to 300W, and the operating frequency is 20 to 40kHz. The tangential flow filtration unit is equipped with a hollow fiber membrane or flat sheet membrane with a molecular weight cutoff of 100-500kDa, and the membrane material is polyethersulfone or regenerated cellulose.

[0022] Preferably, the adaptive chromatography purification module includes:

[0023] A series of chromatography columns connected in sequence, wherein the chromatography column group includes at least an affinity chromatography column and a molecular sieve chromatography column, and may optionally be equipped with an ion exchange chromatography column;

[0024] An online detection unit is provided at the injection end and elution end of the chromatography column assembly, the online detection unit including a UV-Vis spectroscopy detector and a dynamic light scattering detector;

[0025] An electrically operated diversion valve is provided at the elution end of the chromatography column assembly, and the electrically operated diversion valve is electrically connected to the central intelligent control system.

[0026] The UV-Vis spectrometer has a monitoring wavelength range of 220-300 nm and is used to detect the protein absorption peak of the eluent; the DLS detector has a particle size detection range of 10-1000 nm and is used to detect the particle size distribution of particles in the eluent.

[0027] Preferably, the central intelligent control system includes:

[0028] The data acquisition module is used to receive temperature, pH, dissolved oxygen, cell state, eluent spectrum and particle size distribution data from each module;

[0029] The process database contains a dataset of process parameters for at least 50 batches of qualified oncolytic virus production, including transfection conditions, culture parameters, and purification and elution procedures.

[0030] The algorithm processing module adopts a fusion algorithm of random forest and gradient boosting tree, and dynamically adjusts the transfection voltage, chromatography elution flow rate and the timing of the split valve opening and closing based on real-time data and process database.

[0031] The data recording module is used to store the entire process data for each batch of production, with a storage time of ≥5 years.

[0032] Preferably, the microfluidic electroporation unit includes:

[0033] The microfluidic chip has 3-5 parallel microchannels with a channel width of 50-200μm and a depth of 20-50μm.

[0034] Pulse power supply, output voltage range 100-800V, pulse width 10-100μs, pulse count 1-5 times;

[0035] The microfluidic chip's inlet is connected to the plasmid solution storage tank and the packaged cell suspension storage tank, while its outlet is connected to the bioreactor.

[0036] (III) Beneficial Effects

[0037] This invention provides a highly efficient and intelligent packaging and purification system for oncolytic viruses. It offers the following advantages:

[0038] 1. This invention designs a fully enclosed integrated flow path, connecting packaging, harvesting, and purification modules through sterile tubing. It integrates AI image recognition with UV-Vis and DLS online detection to achieve intelligent determination of harvesting timing and purified components, eliminating contamination caused by manual intervention. AI detection replaces manual judgment, significantly improving the accuracy of harvesting and purification. Adaptive chromatography accurately captures target viruses, significantly improving purity and recovery rate. Furthermore, machine learning continuously optimizes parameters, reducing batch variations, providing a reliable solution for the large-scale production of clinical-grade oncolytic viruses. Detailed Implementation

[0039] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0040] Example:

[0041] This invention provides an efficient and intelligent packaging and purification system for oncolytic viruses, including an intelligent virus packaging module, a fully enclosed primary harvesting module, an adaptive chromatography purification module, and a central intelligent control system.

[0042] The intelligent virus packaging module is used to achieve efficient transfection of plasmids into packaging cells, virus packaging culture, and to determine the optimal time to harvest the virus by real-time monitoring of cell status.

[0043] The fully enclosed primary harvest module is connected to the intelligent virus packaging module through sterile tubing. It is used to receive virus culture medium and complete cell lysis, virus release, and preliminary clarification of culture medium in a closed environment.

[0044] The adaptive chromatography purification module is connected to the fully enclosed primary harvest module through sterile tubing. It is used to perform multi-step fine purification of the initially clarified crude virus extract and to accurately collect the target virus fraction based on real-time detection data.

[0045] The central intelligent control system is connected to the intelligent virus packaging module, the fully enclosed primary harvesting module, and the adaptive chromatography purification module via wired or wireless communication. It is used to receive real-time operating data from each module and dynamically coordinate and optimize the process parameters of each module through a preset algorithm model.

[0046] The intelligent virus packaging module, the fully enclosed primary harvesting module, and the adaptive chromatography purification module are connected in series with a sanitary diaphragm valve via a sterile quick-connect connector, forming a continuous and closed integrated production flow path.

[0047] The intelligent virus packaging module includes:

[0048] The bioreactor is equipped with online sensors for temperature, pH, and dissolved oxygen concentration.

[0049] An online microscopic imaging system integrated on top of the bioreactor includes an upright microscope module with 20×-40× objectives, a CMOS image sensor, and an AI image recognition unit.

[0050] A transfection unit connected to the bioreactor via sterile tubing; the transfection unit is a microfluidic electrotransfection unit or a high-throughput nanomaterial transfection unit.

[0051] The AI ​​image recognition unit is used to analyze cell density, fusion degree and lesion effect in real time, and transmits the analysis results to the central intelligent control system.

[0052] Operation of the intelligent virus packaging module:

[0053] The core task of this module is to achieve efficient transfection of adenovirus packaging plasmids into 293 cells and virus packaging culture. The specific steps are as follows:

[0054] (1) Cell preparation and inoculation

[0055] The 293 cells were revived from cryopreservation and cultured in shake flasks until the cell density reached 5 × 10⁻⁶. 5cells / mL; the cell suspension was transferred to a 50L bioreactor through sterile tubing.

[0056] The central intelligent control system sets the following cultivation parameters: temperature 37℃±0.1℃, pH 7.2±0.05, dissolved oxygen concentration 50%±2%, stirring speed 50rpm, and aeration rate 0.5vvm.

[0057] After 60 hours of culture, the online microscopic imaging system begins to acquire cell images in real time. The AI ​​image recognition unit analyzes the cell confluence, and when the confluence reaches 85%, the transfection procedure is triggered.

[0058] (2) Microfluidic electroporation operation

[0059] Preparation of the transfection system: Mix 10 mg of adenovirus packaging plasmid with 2 L293 cell suspension and deliver it to the microfluidic electrotransfection unit using a peristaltic pump;

[0060] The central intelligent control system automatically matches transfection parameters according to cell density: pulse voltage 350V, pulse width 50μs, pulse count 3 times, fluid flow rate 2mL / min;

[0061] The transfected cell suspension is returned to the bioreactor through sterile tubing for further culture.

[0062] (3) Monitoring of virus packaging and determination of harvest timing

[0063] Starting 24 hours post-transfection, the online microscopic imaging system acquired cell images every 30 minutes. The AI ​​image recognition unit analyzed the following indicators using a convolutional neural network algorithm:

[0064] Cell density: calculated from image grayscale values, with an error ≤5%;

[0065] Pathogenic effect: Classified into 0-4 levels according to the degree of cell shrinkage and shedding. When CPE reaches level 3.5 (i.e., more than 75% of cells show pathogenic effects), the AI ​​unit sends a "best harvest" signal to the central control system.

[0066] In this embodiment, a harvest signal is triggered 72 hours after transfection, and the central control system automatically opens the sterile diaphragm valve between the bioreactor and the fully enclosed primary harvest module to transfer the virus culture medium (approximately 40L) to the sterile harvest tank.

[0067] The fully enclosed primary harvesting module includes a sterile harvesting tank, which is connected to the bioreactor of the intelligent virus packaging module via sterile piping.

[0068] The processing unit is integrated inside the aseptic harvesting tank. The processing unit is either an ultrasonic crushing unit or a tangential flow filtration unit.

[0069] The ultrasonic disruption unit has an adjustable power range of 50-300W and an operating frequency of 20-40kHz; the tangential flow filtration unit is equipped with a hollow fiber membrane or flat sheet membrane with a molecular weight cutoff of 100-500kDa, and the membrane material is polyethersulfone or regenerated cellulose.

[0070] This module achieves cell lysis, virus release, and preliminary clarification through tangential flow filtration, with a completely closed-loop operation. The specific steps are as follows:

[0071] (1) Cell lysis: The central control system controls the agitator in the sterile harvesting tank to rotate at 30 rpm. At the same time, the culture medium is circulated between the harvesting tank and the membrane module through the circulation pump of the TFF unit. The transmembrane pressure is controlled at 1.5 bar. The cells are gently lysed by shear force to release virus particles.

[0072] (2) Preliminary clarification and concentration: The "cross-flow filtration" mode is adopted. The permeate (containing cell debris and small molecule impurities) is discharged to the waste tank through the membrane module, and the retained liquid (containing viruses) is returned to the harvest tank.

[0073] (3) The central control system monitors the volume of the retention liquid in real time. When the volume is concentrated to 8L (concentration factor 5 times), it switches to the "dialysis filtration" mode, adds 16L of PBS buffer (pH 7.4) to the harvest tank, and continues to circulate and filter to remove residual culture medium components.

[0074] (4) After clarification, the central control system opens the sterile valve between the harvest tank and the adaptive chromatography purification module to deliver the crude virus extract (about 10L) to the chromatography loading tank.

[0075] After this step, sampling and testing showed that the cell debris removal rate in the crude virus extract was ≥98%, and the virus recovery rate was ≥95%.

[0076] The adaptive chromatography purification module includes:

[0077] A series of chromatography columns connected in sequence, the chromatography column group includes at least an affinity chromatography column and a molecular sieve chromatography column, and may optionally be equipped with an ion exchange chromatography column;

[0078] An online detection unit is located at the injection and elution ends of the chromatography column. The online detection unit includes an ultraviolet-visible spectroscopy detector and a dynamic light scattering detector.

[0079] An electric diversion valve is installed at the elution end of the chromatography column assembly and is electrically connected to the central intelligent control system.

[0080] The UV-Vis spectroscopy detector has a monitoring wavelength range of 220-300 nm and is used to detect the protein absorption peaks in the eluent; the DLS detector has a particle size detection range of 10-1000 nm and is used to detect the particle size distribution in the eluent.

[0081] This module achieves fine virus purification through a two-step process of affinity chromatography followed by molecular sieve chromatography. Operating parameters are dynamically adjusted based on online detection data. The specific steps are as follows:

[0082] (1) Affinity chromatography purification

[0083] Column equilibration: The central control system controls the peristaltic pump to introduce PBS buffer (pH 7.4) into the affinity chromatography column at a flow rate of 10 mL / min until the absorbance of the UV-Vis detector at 280 nm stabilizes (fluctuation ≤ 0.005 AU).

[0084] Sample loading: The crude virus extract was loaded at a flow rate of 8 mL / min. The absorbance at 280 nm was monitored in real time by an online UV-Vis detector. When the absorbance exceeded 0.1 AU, the central control system recorded the loading start point. When the absorbance dropped back to 0.1 AU, the loading was stopped. A total of 10 L of sample was loaded.

[0085] Washing: Pour PBS buffer (pH 7.4) containing 0.5 M NaCl into the filter at a flow rate of 15 mL / min and wash until the absorbance of the UV-Vis detector returns to baseline (≤0.01 AU) to remove non-specifically bound proteins.

[0086] Elution: Switch to elution buffer (containing 0.1M glycine-HCl, pH 2.7), reduce the flow rate to 5 mL / min, and monitor the absorbance at 280 nm in real time with a UV-Vis detector. When the absorbance exceeds 0.05 AU, the central control system controls the electric split valve to switch to "collect" mode to collect the elution peak; when the absorbance drops back to 0.05 AU, the split valve switches to "waste liquid" mode.

[0087] In this step, approximately 200 mL of affinity eluent was collected. Simultaneous monitoring with a DLS detector showed that the particle size in the eluent was concentrated in the range of 90-110 nm (adenovirus characteristic particle size), and the content of impurity particles was ≤2%.

[0088] (2) Molecular sieve chromatography purification

[0089] Column equilibration: PBS buffer (pH 7.4) is bubbled into the molecular sieve chromatography column at a flow rate of 5 mL / min until the absorbance of the UV-Vis detector stabilizes;

[0090] Sample loading: Load the affinity eluent through the sample loop (20 mL) at a flow rate of 5 mL / min;

[0091] Washing and Collection:

[0092] The UV-Vis detector simultaneously monitors the absorbance at 260 nm (nucleic acid characteristic peak) and 280 nm (protein characteristic peak). When a characteristic peak with a 260 nm / 280 nm ratio of 1.3-1.5 (adenovirus characteristic ratio) appears, the DLS detector simultaneously verifies whether the particle size is 90-110 nm.

[0093] When both conditions are met, the central control system controls the diversion valve to open for collection; when the ratio deviates from the range or the particle size is abnormal, collection stops.

[0094] This step collected approximately 50 mL of the target virus fraction. Sampling and testing showed that the content of impurity proteins was ≤0.1% and the virus purity was ≥99%.

[0095] The central intelligent control system includes:

[0096] The data acquisition module is used to receive temperature, pH, dissolved oxygen, cell state, eluent spectrum and particle size distribution data from each module;

[0097] The process database contains a dataset of process parameters for at least 50 batches of qualified oncolytic virus production, including transfection conditions, culture parameters, and purification and elution procedures.

[0098] The algorithm processing module adopts a fusion algorithm of random forest and gradient boosting tree, and dynamically adjusts the transfection voltage, chromatography elution flow rate and the timing of the split valve opening and closing based on real-time data and process database.

[0099] The data recording module is used to store the entire process data for each batch of production, with a storage time of ≥5 years.

[0100] In this embodiment, the core function of the central intelligent control system is reflected in the following three aspects:

[0101] Real-time parameter adjustment:

[0102] Packaging stage: When the online microscopic imaging system detects a cell density growth rate of less than 0.02 × 10⁻⁶, 6 When cells are in use, the system automatically increases the dissolved oxygen concentration from 50% to 60% and the stirring speed from 50 rpm to 60 rpm to ensure the metabolic needs of cells are met.

[0103] Purification stage: Based on the slope of the UV-Vis absorbance curve during the affinity chromatography loading stage, the system determines that the virus concentration in the crude virus extract is 1×10¹. 0 The PFU / mL elution rate automatically reduces the elution flow rate from the preset 8 mL / min to 5 mL / min, extending the binding time between the virus and the packing material and improving elution efficiency.

[0104] Process data record:

[0105] The system automatically records over 1200 parameter points throughout the entire process, including: transfection voltage, culture temperature, CPE occurrence time, TFF concentration factor, and elution peak area. The data is stored in a MySQL database with a retention period of 6 years and can be traced at any time through the system interface.

[0106] Machine learning self-optimization:

[0107] After this batch of production is completed, the system compares the actual production data (such as a virus recovery rate of 85% and a purity of 99.2%) with historical data from 80 batches in the process database, and optimizes the parameters for the next batch using a random forest-gradient boosting tree model.

[0108] The transfection pulse width was adjusted from 50 μs to 45 μs (to reduce cell damage);

[0109] The affinity chromatography elution flow rate was increased from 15 mL / min to 18 mL / min (to shorten the purification time).

[0110] After optimization, the virus recovery rate of the next batch was increased to 90%, and the purification cycle was shortened by 10%.

[0111] The microfluidic electroporation unit includes:

[0112] The microfluidic chip has 3-5 parallel microchannels with a channel width of 50-200μm and a depth of 20-50μm.

[0113] Pulse power supply, output voltage range 100-800V, pulse width 10-100μs, pulse count 1-5 times;

[0114] The microfluidic chip's inlet end is connected to a plasmid solution storage tank and a packaged cell suspension storage tank, while its outlet end is connected to a bioreactor.

[0115] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A highly efficient and intelligent packaging and purification system for oncolytic viruses, characterized in that, It includes an intelligent virus packaging module, a fully enclosed primary harvesting module, an adaptive chromatography purification module, and a central intelligent control system; The intelligent virus packaging module is used to achieve efficient transfection of plasmids into packaging cells, virus packaging culture, and to determine the optimal time to harvest the virus by real-time monitoring of cell status. The fully enclosed primary harvest module is connected to the intelligent virus packaging module through sterile tubing and is used to receive virus culture medium, complete cell lysis, virus release and preliminary clarification of culture medium in a closed environment; The adaptive chromatography purification module is connected to the fully enclosed primary harvest module through sterile tubing. It is used to perform multi-step fine purification of the initially clarified crude virus extract and to accurately collect the target virus fraction based on real-time detection data. The central intelligent control system is connected to the intelligent virus packaging module, the fully enclosed primary harvesting module, and the adaptive chromatography purification module via wired or wireless communication, respectively. It is used to receive real-time operating data from each module and dynamically coordinate and optimize the process parameters of each module through a preset algorithm model. The intelligent virus packaging module, the fully enclosed primary harvesting module, and the adaptive chromatography purification module are connected in series with a sanitary diaphragm valve via a sterile quick-connect connector, forming a continuous and closed integrated production flow path.

2. The oncolytic virus high-efficiency intelligent packaging and purification system according to claim 1, characterized in that: The intelligent virus packaging module includes: The bioreactor is equipped with online sensors for temperature, pH, and dissolved oxygen concentration. An online microscopic imaging system integrated on top of the bioreactor includes an upright microscope module with 20×-40× objectives, a CMOS image sensor, and an AI image recognition unit. A transfection unit connected to the bioreactor via sterile tubing, wherein the transfection unit is a microfluidic electrotransfection unit or a high-throughput nanomaterial transfection unit; The AI ​​image recognition unit is used to analyze cell density, fusion degree and lesion effect in real time, and transmits the analysis results to the central intelligent control system.

3. The oncolytic virus high-efficiency intelligent packaging and purification system according to claim 1, characterized in that: The fully enclosed primary harvesting module includes: A sterile harvesting tank is connected to the bioreactor of the intelligent virus packaging module via sterile piping. A processing unit integrated inside the aseptic harvesting tank, wherein the processing unit is an ultrasonic crushing unit or a tangential flow filtration unit; The power of the ultrasonic disruption unit is adjustable from 50 to 300W, and the operating frequency is 20 to 40kHz. The tangential flow filtration unit is equipped with a hollow fiber membrane or flat sheet membrane with a molecular weight cutoff of 100-500kDa, and the membrane material is polyethersulfone or regenerated cellulose.

4. The oncolytic virus high-efficiency intelligent packaging and purification system according to claim 1, characterized in that: The adaptive chromatography purification module includes: A series of chromatography columns connected in sequence, wherein the chromatography column group includes at least an affinity chromatography column and a molecular sieve chromatography column, and may optionally be equipped with an ion exchange chromatography column; An online detection unit is provided at the injection end and elution end of the chromatography column assembly, the online detection unit including a UV-Vis spectroscopy detector and a dynamic light scattering detector; An electrically operated diversion valve is provided at the elution end of the chromatography column assembly, and the electrically operated diversion valve is electrically connected to the central intelligent control system. The UV-Vis spectrometer has a monitoring wavelength range of 220-300 nm and is used to detect the protein absorption peak of the eluent; the DLS detector has a particle size detection range of 10-1000 nm and is used to detect the particle size distribution of particles in the eluent.

5. The oncolytic virus high-efficiency intelligent packaging and purification system according to claim 1, characterized in that: The central intelligent control system includes: The data acquisition module is used to receive temperature, pH, dissolved oxygen, cell state, eluent spectrum and particle size distribution data from each module; The process database contains a dataset of process parameters for at least 50 batches of qualified oncolytic virus production, including transfection conditions, culture parameters, and purification and elution procedures. The algorithm processing module adopts a fusion algorithm of random forest and gradient boosting tree, and dynamically adjusts the transfection voltage, chromatography elution flow rate and the timing of the split valve opening and closing based on real-time data and process database. The data recording module is used to store the entire process data for each batch of production, with a storage time of ≥5 years.

6. The oncolytic virus high-efficiency intelligent packaging and purification system according to claim 2, characterized in that: The microfluidic electroporation unit includes: The microfluidic chip has 3-5 parallel microchannels with a channel width of 50-200μm and a depth of 20-50μm. Pulse power supply, output voltage range 100-800V, pulse width 10-100μs, pulse count 1-5 times; The microfluidic chip's inlet is connected to the plasmid solution storage tank and the packaged cell suspension storage tank, while its outlet is connected to the bioreactor.