A method and system for producing a high activity immunoglobulin liquid milk

Abstract

The present application relates to the technical field of liquid milk production and processing, and discloses a production method and system of high-activity immunoglobulin liquid milk, which comprises the following steps: screening qualified raw milk and performing milk clarification and defatting through enzyme-linked immunosorbent assay combined with high-performance liquid chromatography, obtaining defatted milk by dynamically adjusting the centrifugal speed and storing the defatted milk at low temperature, periodically detecting the defatted milk by dynamically regulating the storage temperature, and obtaining qualified defatted milk; extracting immunoglobulin, regulating the subtype ratio of the immunoglobulin to adapt to special medical scenarios, and then performing glycosylation modification and desensitization, obtaining immunoglobulin modified liquid and sterile substrate through synergistic sterilization treatment; dynamically adjusting the speed for mixing and correcting the physicochemical properties, performing sterile filling and sealing after optimizing the sensory quality, comprehensively detecting the packaged product, and obtaining qualified finished products; dynamically regulating cold chain parameters according to the activity of immunoglobulin, monitoring the whole process, classifying and stacking, optimizing the adaptability of the production system, treating production wastewater, and recycling residual immunoglobulin.

Description

Technical Field

[0001] This invention relates to the field of liquid milk production and processing technology, specifically to a method and system for producing highly active immunoglobulin liquid milk. Background Technology

[0002] Highly active immunoglobulin liquid milk, due to its immunomodulatory function, is widely applicable to various scenarios including infants, special medical applications, and general functional applications, leading to a growing market demand. Current technologies primarily employ conventional raw milk processing, immunoglobulin extraction, and liquid milk preparation processes to attempt to combine immunoglobulins with liquid milk. However, limitations in process precision, scenario adaptability, and process control make it difficult to simultaneously achieve optimal product activity, purity, safety, and the demands of large-scale production. In existing technologies, raw milk acceptance only tests for immunoglobulin content, not activity and allergens; the centrifugation temperature of defatted milk is too high, leading to immunoglobulin denaturation; the cold chain for raw milk storage and transportation is prone to breakage, and prolonged storage time accelerates immunoglobulin degradation; the extraction process has poor selectivity, resulting in significant loss of activity; desensitization treatment is not performed for infants and young children, resulting in high allergenicity and poor digestibility and absorption; the ratio of IgG, IgA, and IgM subtypes cannot be precisely controlled in special medical scenarios, and the extraction process is complex; the substrate has poor compatibility with immunoglobulins, resulting in an unbalanced nutritional ratio and neglecting the synergistic effect with excipients; different scenarios... The product suffers from several issues: insufficient nutritional compatibility; a single sterilization process that fails to balance sterilization effectiveness with the preservation of immunoglobulin activity; uneven mixing of immunoglobulins with liquid milk, leading to localized high concentrations and potential aggregation and precipitation; significant loss of activity during mixing; insufficient cleanliness of the filling environment, increasing the risk of secondary contamination; poor barrier properties of packaging materials resulting in oxidative inactivation of immunoglobulins; lack of end-point monitoring in the cold chain during storage and transportation, leading to large temperature and humidity fluctuations and short product shelf life; poor compatibility between the production system and large-scale dairy production line equipment, resulting in mismatched capacity between systems and potential production bottlenecks; and insufficient process control precision, leading to large batch-to-batch quality fluctuations. Therefore, there is a need to provide a method and system for producing highly active immunoglobulin liquid milk. Summary of the Invention

[0003] The purpose of this invention is to provide a method and system for producing highly active immunoglobulin liquid milk. To solve the above-mentioned problems in the prior art, this invention achieves this through the following technical solution: The first part, an embodiment of the present invention, provides a method for producing a highly active immunoglobulin liquid milk, which specifically includes the following steps: Step 1: Qualified raw milk is screened by a combination of enzyme-linked immunosorbent assay (ELISA) and high performance liquid chromatography (HPLC), and then the milk is purified and defatted. The centrifugation speed is dynamically adjusted to obtain defatted milk, which is then stored at low temperature. The storage temperature is dynamically controlled and tested regularly to obtain qualified defatted milk. Step 2: After microfiltration pretreatment, the obtained skim milk is used to extract immunoglobulins by molecular imprinting affinity chromatography. The subtype ratios are adjusted to suit specific medical scenarios. Then, glycosylation modification and desensitization are performed. Qualified colostrum is selected to prepare liquid milk base. Through synergistic sterilization, immunoglobulin modified liquid and sterile base are obtained. Step 3: After diluting the immunoglobulin modification solution, mix it with a sterile substrate, dynamically adjust the mixing speed and correct the physicochemical properties, optimize the sensory quality, and then perform aseptic filling and sealing. Conduct comprehensive testing on the packaged product to obtain a qualified finished product. Step 4: The qualified finished products are dynamically adjusted according to the immunoglobulin activity, and the cold chain parameters are monitored throughout the process. They are then sorted and stacked to optimize the adaptability of the production system, treat the production wastewater, and recover residual immunoglobulins.

[0004] The second part, an embodiment of the present invention, provides a production system for highly active immunoglobulin liquid milk, which specifically includes the following modules: Screening and processing module: Qualified raw milk is screened by a combination of enzyme-linked immunosorbent assay (ELISA) and high performance liquid chromatography (HPLC), and then the milk is purified and defatted. The centrifugation speed is dynamically adjusted to obtain defatted milk, which is then stored at low temperature. The storage temperature is dynamically controlled and tested regularly to obtain qualified defatted milk. Modification and preparation module: The obtained skim milk is pretreated by microfiltration and then immunoglobulins are extracted by molecular imprinting affinity chromatography. The subtype ratios are adjusted to suit special medical scenarios. Then, glycosylation modification and desensitization are performed. Qualified colostrum is selected to prepare liquid milk base. Through synergistic sterilization treatment, immunoglobulin modified liquid and sterile base are obtained. Mixed Preservation Module: After diluting the immunoglobulin modified solution, it is mixed with a sterile substrate. The mixing speed is dynamically adjusted and the physicochemical properties are corrected. After optimizing the sensory quality, it is aseptically filled and sealed. The packaged product is then subjected to comprehensive testing to obtain a qualified finished product. The detection and adaptation module dynamically adjusts cold chain parameters based on immunoglobulin activity and monitors the entire process of obtaining qualified finished products. It also classifies and stacks the products, optimizes the compatibility of the production system, treats production wastewater, and recovers residual immunoglobulins.

[0005] The beneficial effects of this invention are: 1. By combining colostrum selection, immunoglobulin activity, and allergen detection, the problem of only testing the content of raw milk without testing its activity and allergens is solved; low-temperature defatting and dynamic speed control are used to solve the activity loss caused by excessively high centrifugation temperatures; a dynamic low-temperature storage formula and full-process cold chain monitoring are designed to adapt to the purity requirements of raw materials in special scenarios; molecular imprinted affinity chromatography extraction is used to improve purity and recovery rate, solving the problems of poor selectivity, low purity, and low recovery rate in extraction processes; DBD plasma-assisted glycosylation modification is used to reduce allergenicity, decrease molecular weight, and improve the intestinal absorption efficiency of infants; a subtype regulation formula is designed to precisely regulate the ratio of IgG, IgA, and IgM to prevent imbalance. 2. A dynamic excipient addition formula is adopted to achieve synergistic effects between immunoglobulins and excipients. Nutrient components are precisely controlled for different scenarios. Pasteurization and PEF sterilization are used in synergistic processes, with sterilization parameters dynamically adjusted to balance sterilization effectiveness and activity retention, adapting to industrial production capacity requirements. A mixing ratio control formula and dilution pretreatment are designed, combined with dynamic mixing speed to solve the problems of uneven mixing and aggregation / precipitation caused by excessively high local concentrations. Mixing temperature and speed are controlled to address activity loss during mixing. Two-stage dynamic homogenization parameter control is used to balance fat globule breakage and activity retention. Multiple sterilization processes are employed to address insufficient filling cleanliness and secondary contamination. A dynamic cold chain temperature formula and IoT-based full-process monitoring are designed. Standardized interfaces and dynamic capacity control are used to construct a closed-loop process control system, reducing the batch-to-batch quality variation coefficient. Attached Figure Description

[0006] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0007] Figure 1 This is a flowchart of the steps in a method for producing a highly active immunoglobulin liquid milk according to Embodiment 1 of the present invention; Figure 2 This is a schematic diagram of the production system for a highly active immunoglobulin liquid milk provided in Embodiment 2 of the present invention. Detailed Implementation

[0008] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. 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 should fall within the scope of protection of the present invention.

[0009] Example 1: As Figure 1 As shown in the figure, the method for producing a highly active immunoglobulin liquid milk provided by an embodiment of the present invention specifically includes the following steps: Step 1: Qualified raw milk is screened by a combination of enzyme-linked immunosorbent assay (ELISA) and high performance liquid chromatography (HPLC), and then the milk is purified and defatted. The centrifugation speed is dynamically adjusted to obtain defatted milk, which is then stored at low temperature. The storage temperature is dynamically controlled and tested regularly to obtain qualified defatted milk. In a specific embodiment, colostrum from healthy dairy cows within 72 hours postpartum is selected as the raw milk. The raw milk is required to have a fat content between 3.2% and 3.8%, a protein content between 3.0% and 3.5%, and a total bacterial count of less than or equal to 100 CFU / mL. Milk from cows suffering from mastitis or metabolic diseases is excluded. This avoids the impact of harmful substances and abnormal proteins in the raw milk on the subsequent immunoglobulin activity from the source. At the same time, it is suitable for the high purity requirements of raw materials for infants and special medical scenarios, reducing the risk of subsequent sensitization and potential loss of activity. Joint detection of immunoglobulin activity and allergens: Enzyme-linked immunosorbent assay (ELISA) combined with high performance liquid chromatography (HPLC) was used to simultaneously obtain the immunoglobulin activity, immunoglobulin content and allergen content in the raw milk. Specifically, 1 mL of raw milk sample was taken, and immunoglobulin-specific coated antigen from the ELISA kit was added. The mixture was incubated at the preset standard temperature for 60 min, washed three times, and then enzyme-labeled secondary antibody was added. The mixture was incubated at the preset standard temperature for 30 min, washed again, and then substrate solution was added. The mixture was reacted at room temperature in the dark for 15 min, and then stop solution was added. The absorbance was detected using a microplate reader at 450 nm. A blank control group and a standard control group were also set up. The allergen content in the raw milk was detected using HPLC, and the peak area of ​​the allergen was recorded. A standard control group for allergens was also set up to obtain the peak area and concentration of the standard. For example, a C18 column was selected, the mobile phase was acetonitrile-0.1% trifluoroacetic acid solution, the flow rate was 1.0 mL / min, the detection wavelength was 280 nm, the column temperature was 30 °C, and the injection volume was 20 μL. By combining the test results of blank control and standard control, the actual activity of immunoglobulins in raw milk is obtained. At the same time, by combining the allergen standard and the allowable concentration, the allergen adaptation adjustment coefficient is determined. The allergen adaptation adjustment coefficient corrects the immunoglobulin activity test results and can also be directly used as the parameter basis for subsequent desensitization treatment. If the actual immunoglobulin activity is greater than or equal to 80 U / mL and the allergen adaptation adjustment factor is greater than or equal to 0.8, the raw milk is qualified; if the actual immunoglobulin activity is less than 80 U / mL, the raw milk is directly rejected; if the allergen adaptation adjustment factor is less than 0.8, the raw milk is retained and labeled. Qualified raw milk is fed into a low-temperature milk purifier, and the milk purifying temperature and centrifugation speed are controlled. The milk purifying temperature and centrifugation speed are dynamically adjusted according to the actual activity of immunoglobulins in the raw milk. The higher the immunoglobulin activity, the higher the centrifugation speed, to ensure thorough milk purifying and remove tiny impurities and bacterial cells from the raw milk. If immunoglobulin activity decreases, reduce the centrifugation speed to reduce the shear force that damages the immunoglobulin molecular structure and avoid loss of activity; After defatting the milk, the upper fat layer is removed, and the defatted milk is retained. The fat layer is collected separately for by-product processing to avoid the influence of fat on the purity of immunoglobulin extraction. Skim milk is placed in a sterile, low-temperature storage tank, and the storage temperature is dynamically adjusted according to the actual activity of immunoglobulins. If the immunoglobulin activity is higher than 80 U / mL, the storage temperature should be lowered to further inhibit immunoglobulin degradation and inhibit microbial growth. If the immunoglobulin activity is equal to the preset activity threshold, maintain the basic storage temperature; if the immunoglobulin activity is less than 80 U / mL, increase the storage temperature to avoid skim milk coagulation caused by low temperature. During storage, the immunoglobulin activity and bacterial count of skim milk were tested every 2 hours. When the activity decrease rate exceeded 1% / h or the bacterial count exceeded 100 CFU / mL, dynamic adjustments were made immediately. Step 2: After microfiltration pretreatment, the obtained skim milk is used to extract immunoglobulins by molecular imprinting affinity chromatography. The subtype ratios are adjusted to suit specific medical scenarios. Then, glycosylation modification and desensitization are performed. Qualified colostrum is selected to prepare liquid milk base. Through synergistic sterilization, immunoglobulin modified liquid and sterile base are obtained. In a specific embodiment, the obtained skim milk was used for immunoglobulin extraction using a molecularly imprinted affinity chromatography column. The specific operation and data processing procedure are as follows: Skim milk was pretreated through a 0.22 μm ceramic microfiltration membrane. The microfiltration temperature was controlled between 4℃ and 8℃. The microfiltration pressure was dynamically adjusted according to the actual activity of immunoglobulins in the skim milk. The higher the immunoglobulin activity, the higher the microfiltration pressure was, to ensure that impurities such as small molecule peptides, lactose, and inorganic salts were completely removed, while avoiding excessive pressure that would cause immunoglobulin molecules to be adsorbed on the membrane surface, thus reducing activity loss. After microfiltration, the crude extract containing immunoglobulins was collected, and the retained impurities were discarded. The crude extract is fed into a molecularly imprinted affinity chromatography column. The column packing material is an immunoglobulin-specific molecularly imprinted polymer. Phosphate buffer is used as the elution buffer. The elution flow rate is dynamically adjusted according to the microfiltration pressure. The higher the microfiltration pressure, the more thoroughly the impurities are removed, and the higher the elution flow rate, thus improving the extraction efficiency and meeting the needs of large-scale industrial production. If the microfiltration pressure decreases, the elution flow rate is reduced to ensure that the immunoglobulins are fully bound to the packing material before being eluted, thereby improving the extraction purity. During the chromatography process, the absorbance of the eluent is monitored in real time. If the absorbance reaches the peak value, the immunoglobulin extract is collected. If the absorbance drops to 10% of the peak value, the collection is stopped, and the extraction is completed. High performance liquid chromatography (HPLC) was used to detect the purity and recovery rate of immunoglobulins in the purified extract. The mass of immunoglobulins and impurities was obtained by detecting the chromatographic peak area of ​​immunoglobulins and impurities in the purified extract and combining it with the peak area of ​​immunoglobulin standards. A preset impurity correction coefficient was used to ensure the accuracy of the calculation results. The impurity correction coefficient was dynamically adjusted according to the elution flow rate. The higher the elution flow rate, the larger the correction coefficient, so as to avoid impurity residue due to excessively fast elution flow rate. To address the imbalance in immunoglobulin subtype ratios in specific medical settings, subtype regulation was performed on the purified extract. The specific operation and data processing procedures are as follows: The contents of IgG, IgA and IgM in the pure extract were detected by ELISA. The target concentration of each subtype was determined according to the needs of special medical scenarios. The volume of the corresponding subtype stock solution was calculated. The corresponding deficient immunoglobulin subtype extracted from the special subtype stock solution was added to the pure extract. After adjustment, the total immunoglobulin concentration was maintained between [1000 mg / L and 3000 mg / L]. After the adjustment is completed, the proportion of subtypes is detected again using the ELISA method to ensure that the proportion error is controlled within ±5%, so as to meet the specific needs of immunoglobulin subtypes in special medical scenarios. To address the shortcomings of high immunoglobulin allergenicity and poor digestion and absorption in infants and young children, the purified extract was modified by dielectric barrier discharge plasma-assisted glycosylation. For example, glucose is added to the purified extract after conditioning. The amount of glucose added is [10%, 15%] of the immunoglobulin mass. After mixing evenly, the pH of the system is adjusted to [7.0, 7.5], the temperature is controlled between [30℃, 40℃], the stirring speed is 50 r / min, and the stirring time is 10 min to ensure that the glucose and immunoglobulin are fully mixed. The pretreated system is fed into a DBD plasma treatment device. The modification time and plasma power are dynamically adjusted according to the sensitizer adaptation adjustment coefficient obtained in step 1. The smaller the sensitizer adaptation adjustment coefficient, the longer the modification time and the higher the plasma power, to ensure that the sensitizing epitopes of immunoglobulins are fully blocked and the sensitization is reduced. The larger the sensitizer adaptation adjustment coefficient, the shorter the modification time and the lower the plasma power, to avoid excessive modification that could lead to loss of immunoglobulin activity. During the modification process, the sensitization reduction rate of the system is monitored in real time. The sensitization reduction rate is obtained by detecting the absorbance of the allergen before and after modification. When the sensitization reduction rate is ≥80%, the modification is stopped. After modification, the immunoglobulin molecules are glycosylated, reducing the molecular weight to 80-120kDa. The intestinal absorption efficiency of infants and young children is increased to more than 60% of that of adults, which is far higher than the 30% of the existing technology. At the same time, the sensitization rate is reduced by ≥80%, solving the core defects in the infant and young child scenario. To meet the nutritional needs of different scenarios, a liquid emulsion base adapted to immunoglobulins is prepared to avoid the base components affecting the activity of immunoglobulins, while achieving synergistic nutrition. Select qualified unskimmed raw milk as the base ingredient, and add excipients according to different needs: for infants and young children, add galactooligosaccharides, DHA and ARA; for special medical needs, add lactoferrin, vitamin C and zinc; for general functional needs, add probiotics and dietary fiber. The amount of excipients added is dynamically adjusted according to the actual activity of immunoglobulins in the immunoglobulin modification solution. The higher the activity of immunoglobulins, the higher the amount of excipients added, so as to achieve the synergistic effect of immunoglobulins and excipients. For example, prebiotics promote the growth of beneficial bacteria in the gut and improve the absorption efficiency of immunoglobulins; lactoferrin enhances the immunomodulatory effect. When immunoglobulin activity decreases, reduce the amount of excipients added to avoid excessive excipients causing abnormal substrate viscosity. Feed the main material and excipients into a high-speed mixer, control the mixing temperature between [10℃ and 15℃], the mixing speed at 2000 r / min, and the mixing time at 15 min to ensure that the excipients are evenly dispersed in the substrate. The nutritional composition of the mixed base is tested and precisely adjusted for different scenarios: for infants and young children, the fat content is controlled between [3.0%, 3.2%], the protein content between [2.8%, 3.0%], and the lactose content between [4.5%, 5.0%]. If the fat content is too high, skim milk is added to adjust it. The volume of skim milk to be added is calculated based on the difference between the actual fat content of the base and the target fat content. In special medical settings, the fat content is controlled between [2.0%, 2.5%], the protein content between [3.5%, 4.0%], and the lactose content between [3.0%, 3.5%]. If the protein content is insufficient, whey protein is added to adjust it. The expected amount of whey protein to be added is calculated based on the difference between the actual protein content of the base and the target protein content. A combined pasteurization and pulsed electric field sterilization process is employed. The regulated substrate is fed into a plate heat exchanger. The pasteurization temperature and time are dynamically adjusted based on the actual activity of immunoglobulins in the immunoglobulin modification solution. Higher immunoglobulin activity requires a higher pasteurization temperature and shorter time to ensure sterilization effectiveness while minimizing heat damage. Conversely, if immunoglobulin activity decreases, the sterilization temperature is lowered and the sterilization time is extended to prevent over-sterilization and further loss of immunoglobulin activity. After pasteurization, the substrate is immediately cooled to [10℃, 15℃] to prevent the loss of nutrients and immunoglobulin activity due to prolonged high temperatures. The cooled substrate is then fed into a PEF sterilization device. The PEF electric field strength and pulse frequency are dynamically adjusted according to the actual activity of immunoglobulins in the immunoglobulin modification solution. The higher the immunoglobulin activity, the higher the electric field strength and pulse frequency, which fully destroys the heat-resistant spore cell membrane and ensures the sterilization effect. If the immunoglobulin activity decreases, the electric field strength and pulse frequency are reduced to avoid the shear force generated by the high electric field strength from damaging the immunoglobulin molecular structure. Step 3: After diluting the immunoglobulin modification solution, mix it with a sterile substrate, dynamically adjust the mixing speed and correct the physicochemical properties, optimize the sensory quality, and then perform aseptic filling and sealing. Conduct comprehensive testing on the packaged product to obtain a qualified finished product. In a specific embodiment, the immunoglobulin modification solution is diluted and mixed with a sterile substrate in a certain proportion. The actual addition volume of the immunoglobulin modification solution is obtained based on the total target immunoglobulin concentration for different scenarios, combined with the concentration of autoimmune globulins in the sterile substrate and the concentration of immunoglobulins in the immunoglobulin modification solution. After the calculation is completed, the immunoglobulin modified solution is diluted to the preset ratio of the original concentration using sterile phosphate buffer. During the dilution process, the ambient temperature is controlled at a low temperature to avoid the impact of temperature fluctuations on the activity of immunoglobulins. The dilution operation follows aseptic procedures throughout to prevent contamination by other microorganisms. After dilution, slowly and evenly add the immunoglobulin modified solution to the sterile substrate, keeping the mixture stirred during the addition process to avoid uneven mixing caused by adding a large amount at once. A twin-shaft paddle mixer is used for mixing. The ambient temperature is controlled at a low temperature during the mixing process. The mixing time is adjusted according to the actual mixing effect. The mixing speed is dynamically controlled according to the immunoglobulin concentration in the diluted immunoglobulin modified solution. If the concentration increases after dilution, the speed is increased to ensure that the immunoglobulin modified solution is fully mixed with the sterile substrate. If the concentration decreases, the speed is decreased. During the mixing process, an online monitoring system is used to detect the uniformity of the mixing system in real time. The concentration of immunoglobulins at different locations is detected by multi-point sampling, and the average value and standard deviation of the concentration at each sampling point are calculated to evaluate the mixing uniformity. If the detection shows that the mixing is not uniform, the mixing speed and mixing time are adjusted in time until the mixing system reaches a uniform state. After mixing, the mixture system was comprehensively tested, including immunoglobulin activity, concentration, homogeneity and physicochemical properties. The immunoglobulin activity and concentration were tested using the same methods as before. Homogeneity was evaluated using the coefficient of variation of concentration. The physicochemical properties were mainly tested for pH and viscosity of the system. If the pH value is abnormal, use sterile citric acid or sodium hydroxide solution to adjust it. Based on the total volume of the mixed system and the difference between the actual pH value and the target pH value, obtain the volume of the adjusting solution to be added. Slowly add the adjusting solution and stir continuously to ensure that the pH value is adjusted to the preset pH value range. If the viscosity is too high or too low, it can be adjusted by adding sterile water or concentrated base. After the mixed system passes the test, it is sent to an intelligent homogenizer for homogenization. A two-stage homogenization process is adopted. During the homogenization process, the ambient temperature is controlled at the preset temperature. The pressure of the first-stage homogenization and the pressure of the second-stage homogenization are dynamically adjusted according to the actual activity of immunoglobulins in the mixed system. If the immunoglobulin activity increases, the homogenization pressure is increased. If the activity decreases, the homogenization pressure is reduced. During the homogenization process, the immunoglobulin activity and fat globule diameter of the mixed system are monitored in real time, and the homogenization pressure is adjusted in a timely manner based on the monitoring results. After homogenization, the mixture is sent to the aseptic filling process. Before filling, the filling environment, filling equipment and packaging materials are pretreated. The filling workshop is adjusted to a cleanliness level, and the temperature and relative humidity in the workshop are controlled within a preset range. The filling equipment includes: filling machine, pipeline and filling head. The filling equipment is thoroughly sterilized, and the filling head is additionally sterilized by soaking in a special sterilization reagent. The packaging materials are sterile and are sterilized by a combination of immersion in sterilizing reagents and ultraviolet irradiation to ensure that the packaging materials are sterile and undamaged. After pretreatment, the homogenized mixture is fed into an aseptic filling machine. The filling speed is dynamically adjusted according to the primary homogenization pressure. During the filling process, the filling volume is strictly controlled to ensure that the filling volume of each package of product is consistent and the filling error is controlled within the preset range. After filling, the packaging is immediately sealed using a heat-sealing process. The heat-sealing temperature, time, and pressure are controlled within the preset range to ensure a tight seal and no leakage. After sealing, the packaged product undergoes comprehensive testing, including leakage detection, sterility testing, and packaging appearance inspection. Leakage detection uses the immersion pressurization method, in which the product is immersed in sterile water, pressure is applied and maintained for a preset period of time, and the presence of bubbles is observed to determine whether the packaging is tightly sealed. The aseptic testing uses the same microbial testing method as the previous one, and samples are taken to test the microbial content in the product. The packaging appearance inspection uses a combination of manual visual inspection and machine inspection to check whether the packaging is damaged, wrinkled, leaked or other abnormalities. Products that fail the test are promptly removed to obtain qualified finished products. Step 4: The qualified finished products are dynamically adjusted according to the immunoglobulin activity, and the cold chain parameters are monitored throughout the process. They are then sorted and stacked to optimize the adaptability of the production system, treat the production wastewater, and recover residual immunoglobulins. In a specific embodiment, qualified finished products are monitored and stored throughout the entire cold chain. Based on the product type and applicable scenario of the qualified finished products, and combined with the actual activity of immunoglobulins in the qualified finished products, the cold chain temperature is dynamically adjusted. If the activity of immunoglobulins increases, the cold chain temperature is reduced to inhibit immunoglobulin degradation and microbial growth. If the activity decreases, the cold chain temperature is increased. Humidity is controlled simultaneously during the cold chain process. Low-temperature products and normal-temperature products adopt corresponding cold chain humidity standards. An Internet of Things cold chain monitoring system is used to monitor all links involving the storage and transportation of qualified finished products, including refrigerated transport vehicles, refrigerated warehouses, distributor storage, and refrigerated cabinets at retail terminals. Temperature and humidity sensors are installed to collect cold chain parameters in real time. The collection frequency is kept constant. The collected temperature and humidity data are transmitted to the monitoring platform in real time. The platform monitors the data in real time. When abnormal fluctuations in temperature or humidity are detected, the monitoring platform immediately issues an early warning signal and automatically adjusts the temperature and humidity of the corresponding refrigeration equipment. The cold chain parameters are quickly adjusted to the target range, and each qualified finished product is affixed with a unique traceability code. The traceability code is associated with the cold chain parameters throughout the entire cold chain process, which facilitates subsequent quality control and problem tracing. When storing qualified finished products, they are sent to the corresponding dedicated warehouse according to product type. Low-temperature products are sent to refrigerated warehouses, and room-temperature products are sent to ambient-temperature warehouses. During storage, qualified finished products are stacked in layers to avoid compressing the packaging, which could cause packaging damage or leakage. The temperature and humidity of the refrigerated warehouse are kept consistent with the cold chain parameters, and the quality of qualified finished products in the warehouse is regularly tested. Ambient temperature warehouses should avoid direct sunlight and high temperature and humidity environments. The temperature and humidity inside the warehouse should be strictly controlled. When qualified finished products are released from the warehouse, the first-in, first-out principle should be followed. Each qualified finished product released from the warehouse should be checked to ensure that the qualified finished products are within their shelf life. Before qualified finished products leave the warehouse, a comprehensive test is conducted again, covering immunoglobulin activity, concentration, subtype ratio, allergenicity, microbial count, physicochemical properties, and packaging integrity. Based on the applicable scenarios of qualified finished products, targeted testing of corresponding items is carried out. For special medical scenarios, the focus is on testing the proportion of immunoglobulin subtypes, and for infant and young children scenarios, the focus is on testing sensitization. Physicochemical property testing ensures that the pH value and viscosity are within the preset viscosity range. Set up blank controls and standard controls to ensure the accuracy and reliability of test results. For finished products that fail the test, rework or destroy them in a timely manner. Based on the total capacity of the production line and the effective volume of the production system, the operating parameters of the capacity are dynamically adjusted to ensure that the capacity of each module is matched. Construct a closed-loop control system for the production process, collect core parameters of each production step in real time, keep the collection frequency fixed, process and analyze the collected parameters in real time, and compare them with the set parameter range; If a parameter is detected to deviate from the set range, the operating parameters of the corresponding equipment are automatically adjusted. The adjustment response is timely to ensure the stability of the production process, reduce quality fluctuations between batches, and fully record all production parameters of each batch of qualified finished products. A certain amount of high-concentration organic wastewater is generated during the production process. The wastewater is treated using a three-stage treatment process. First, the wastewater is sent to a pretreatment tank, the pH value of the wastewater is adjusted to the preset pH value range, flocculant is added for flocculation treatment, and the amount of flocculant added is adjusted according to the actual pollution level of the wastewater. After stirring evenly, the wastewater is allowed to stand, and then filtered to remove suspended impurities, thus completing the wastewater pretreatment. The pretreated wastewater is sent to an ultrafiltration membrane system to recover residual immunoglobulins from the wastewater. The pore size, pressure and temperature of the ultrafiltration membrane are controlled within a preset range to ensure the immunoglobulin recovery effect. The recovered immunoglobulins are purified and modified before being reused in production. The wastewater after immunoglobulin recovery is sent to a biodegradation tank for further treatment. Activated sludge is inoculated into the biodegradation tank, and the temperature, dissolved oxygen, and aeration time in the tank are controlled within a preset range. Through the biological action of the activated sludge, the organic pollutants in the wastewater are degraded until the wastewater meets the national wastewater discharge standards.

[0010] Example 2: As Figure 2 As shown in the figure, the production system for highly active immunoglobulin liquid milk provided in this embodiment of the invention specifically includes the following modules: Screening and processing module: Qualified raw milk is screened by a combination of enzyme-linked immunosorbent assay (ELISA) and high performance liquid chromatography (HPLC), and then the milk is purified and defatted. The centrifugation speed is dynamically adjusted to obtain defatted milk, which is then stored at low temperature. The storage temperature is dynamically controlled and tested regularly to obtain qualified defatted milk. Modification and preparation module: The obtained skim milk is pretreated by microfiltration and then immunoglobulins are extracted by molecular imprinting affinity chromatography. The subtype ratios are adjusted to suit special medical scenarios. Then, glycosylation modification and desensitization are performed. Qualified colostrum is selected to prepare liquid milk base. Through synergistic sterilization treatment, immunoglobulin modified liquid and sterile base are obtained. Mixed Preservation Module: After diluting the immunoglobulin modified solution, it is mixed with a sterile substrate. The mixing speed is dynamically adjusted and the physicochemical properties are corrected. After optimizing the sensory quality, it is aseptically filled and sealed. The packaged product is then subjected to comprehensive testing to obtain a qualified finished product. The detection and adaptation module dynamically adjusts cold chain parameters based on immunoglobulin activity and monitors the entire process of obtaining qualified finished products. It also classifies and stacks the products, optimizes the compatibility of the production system, treats production wastewater, and recovers residual immunoglobulins.

[0011] The above provides a detailed description of one embodiment of the present invention, but the content described is only a preferred embodiment of the present invention and should not be considered as limiting the scope of the present invention. The above formulas are all dimensionless numerical calculations, and the formulas are derived from software simulations based on a large amount of collected data to obtain the most recent real-world situation. The preset parameters in the formulas are set by those skilled in the art based on actual conditions and historical experience, and can be adjusted according to actual conditions. The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. All equivalent changes and improvements made in accordance with the scope of the present invention should still fall within the patent coverage of the present invention.

Claims

1. A method for producing a highly active immunoglobulin liquid milk, characterized in that, Includes the following steps: Qualified raw milk was screened by a combination of enzyme-linked immunosorbent assay (ELISA) and high performance liquid chromatography (HPLC). The milk was then purified and defatted. The centrifugation speed was dynamically adjusted to obtain defatted milk, which was then stored at low temperature. The storage temperature was dynamically controlled and the milk was tested regularly to obtain qualified defatted milk. The obtained skim milk was pretreated by microfiltration and then immunoglobulins were extracted by molecular imprinting affinity chromatography. The proportion of its subtypes was adjusted to suit special medical scenarios. Then, glycosylation modification and desensitization were performed. Qualified colostrum was selected to prepare liquid milk base. Through synergistic sterilization treatment, immunoglobulin modified liquid and sterile base were obtained. After diluting the immunoglobulin modification solution, it was mixed with a sterile substrate. The mixing speed was dynamically adjusted and the physicochemical properties were corrected. After optimizing the sensory quality, it was aseptically filled and sealed. The packaged products were then subjected to comprehensive testing to obtain qualified finished products. The qualified finished products are dynamically adjusted according to the immunoglobulin activity, and the cold chain parameters are monitored throughout the process. They are then sorted and stacked to optimize the adaptability of the production system, treat production wastewater, and recover residual immunoglobulins.

2. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for joint detection is as follows: Colostrum from healthy dairy cows within 72 hours postpartum was selected as raw milk, and its fat content, protein content, and total bacterial count were controlled. Milk from diseased dairy cows was removed. Samples were added to the immunoglobulin-specific coated antigen in the ELISA kit, incubated at the preset standard temperature, washed, and then incubated again with enzyme-labeled secondary antibody. After washing, substrate solution was added and reacted at room temperature in the dark. The absorbance was measured using an enzyme-linked immunosorbent assay (ELISA) reader after adding the stop solution, with blank and standard control groups also included. The allergen content was detected using C18 column HPLC, with an allergen standard control group included. The actual immunoglobulin activity and allergen adaptation adjustment coefficient were obtained by combining the control results. Qualified raw milk was screened, and raw milk with an activity <80U / mL was rejected, while raw milk with an allergen adaptation adjustment coefficient <0.8 was labeled and retained.

3. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for obtaining skim milk is as follows: Qualified raw milk is fed into a low-temperature milk purifier. The milk purifier temperature and centrifugation speed are controlled. The centrifugation speed is dynamically adjusted according to the actual activity of immunoglobulins in the raw milk to ensure thorough milk purification. If the activity is low, the speed is reduced to reduce damage to immunoglobulin molecules. After defatting, the upper fat layer is removed and collected separately for by-product processing, while the defatted milk is retained.

4. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for dynamically adjusting the storage temperature is as follows: Skim milk is placed in a sterile low-temperature storage tank. The storage temperature is dynamically adjusted according to the actual activity of immunoglobulins: if the activity is >80U / mL, the temperature is lowered; if it is equal to the preset activity threshold, the base temperature is maintained; if it is <80U / mL, the temperature is raised to avoid coagulation. During storage, the immunoglobulin activity and bacterial count are tested every 2 hours. If the activity decrease rate is >1% / h or the bacterial count is >100CFU / mL, adjustments are made immediately.

5. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for extracting immunoglobulins is as follows: Skim milk was pretreated with a 0.22 μm ceramic microfiltration membrane. The pressure was dynamically adjusted according to the actual activity of immunoglobulins, with higher pressure for higher activity to retain small molecule impurities. The crude extract containing immunoglobulins after microfiltration was collected, and the retained impurities were discarded. The crude extract was then sent to an immunoglobulin-specific molecularly imprinted polymer-packed chromatography column, with phosphate buffer as the elution buffer. The elution flow rate was dynamically adjusted according to the microfiltration pressure, with the flow rate reduced when the pressure was low to ensure extraction purity. During chromatography, the absorbance of the eluent was monitored in real time. The pure extract was collected starting at the peak and stopped when it dropped to 10% of the peak value. The purity and recovery rate of immunoglobulins in the pure extract were determined by HPLC. The mass of immunoglobulins and impurities was obtained by combining the peak area of ​​the standard. The impurity correction coefficient was dynamically adjusted according to the elution flow rate.

6. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for adapting to special medical scenarios is as follows: The contents of IgG, IgA and IgM in the purified extract were detected by ELISA. The target concentration of each subtype was determined according to the needs of special medical scenarios. The corresponding deficient subtypes in the special subtype stock solution were calculated and added to control the total immunoglobulin concentration after regulation. After regulation, the subtype ratio was detected again using the ELISA method.

7. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for immunoglobulin glycosylation modification and desensitization is as follows: Immunoglobulin glucose was added to the purified extract after subtype regulation. After mixing evenly, the pH and temperature of the system were adjusted. The system was then sent to a DBD plasma treatment device. The modification time and power were dynamically adjusted according to the obtained allergen adaptation adjustment coefficient. The allergen reduction rate was monitored in real time, and the modification was stopped when it dropped to ≥80%.

8. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for the synergistic sterilization process is as follows: Select qualified non-skimmed raw milk as the base ingredient, and add auxiliary ingredients according to the scenario: for infant and toddler scenarios, add galactooligosaccharides, DHA, and ARA; for special medical scenarios, add lactoferrin, vitamin C, and zinc; for general functional scenarios, add probiotics and dietary fiber. The amount of excipients added is dynamically adjusted according to the activity of immunoglobulins in the immunoglobulin modification solution. The main ingredients and excipients are fed into a high-speed mixer, and the nutritional components are tested and adjusted according to the scenario: for infants and young children, fat, protein, and lactose are controlled, and if the fat is too high, skim milk is added to adjust; for special medical scenarios, fat, protein, and lactose are controlled, and if the protein is insufficient, whey protein is added to adjust. The process employs a combination of pasteurization and PEF sterilization. The pasteurization parameters are dynamically adjusted according to the activity of immunoglobulins. After sterilization, the sample is cooled and then fed into the PEF equipment, where the electric field strength and pulse frequency are dynamically adjusted according to the activity of immunoglobulins.

9. The method for producing a highly active immunoglobulin liquid milk according to claim 1, characterized in that, The method for correcting physicochemical properties is as follows: Based on the total immunoglobulin concentration of the target scenario, combined with the immunoglobulin concentration in the sterile substrate and the immunoglobulin modification solution, the actual volume of the modification solution to be added is determined. The modified solution was diluted to the preset ratio using sterile phosphate buffer, with low temperature and aseptic procedures followed during dilution. The diluted modified solution was slowly added to the sterile substrate and stirred continuously. A biaxial paddle mixer was used for low-temperature mixing, with the speed dynamically adjusted according to the concentration of immunoglobulins in the diluted modified solution. The speed was increased when the concentration was high and decreased when the concentration was low. The mixing uniformity is monitored online, and the rotation speed and time are adjusted if it is not up to standard. After mixing, the activity, concentration, uniformity and physicochemical properties of immunoglobulins are tested. If the pH is abnormal, it is adjusted with sterile citric acid or sodium hydroxide solution. If the viscosity is too high or too low, sterile water or concentrated base is added for adjustment.

10. A production system for a highly active immunoglobulin liquid emulsion, the system being used to perform the production method according to any one of claims 1-9, characterized in that, include: Screening and processing module: Qualified raw milk is screened by a combination of enzyme-linked immunosorbent assay (ELISA) and high performance liquid chromatography (HPLC), and then the milk is purified and defatted. The centrifugation speed is dynamically adjusted to obtain defatted milk, which is then stored at low temperature. The storage temperature is dynamically controlled and tested regularly to obtain qualified defatted milk. Modification and preparation module: The obtained skim milk is pretreated by microfiltration and then immunoglobulins are extracted by molecular imprinting affinity chromatography. The subtype ratios are adjusted to suit special medical scenarios. Then, glycosylation modification and desensitization are performed. Qualified colostrum is selected to prepare liquid milk base. Through synergistic sterilization treatment, immunoglobulin modified liquid and sterile base are obtained. Mixed Preservation Module: After diluting the immunoglobulin modified solution, it is mixed with a sterile substrate. The mixing speed is dynamically adjusted and the physicochemical properties are corrected. After optimizing the sensory quality, it is aseptically filled and sealed. The packaged product is then subjected to comprehensive testing to obtain a qualified finished product. The detection and adaptation module dynamically adjusts cold chain parameters based on immunoglobulin activity and monitors the entire process of obtaining qualified finished products. It also classifies and stacks the products, optimizes the compatibility of the production system, treats production wastewater, and recovers residual immunoglobulins.

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