Biopharmaceutical liquid medicine extracting and filtering method and system

Abstract

The invention relates to the technical field of biopharmacy, and discloses a biopharmacy liquid medicine extracting and filtering method and system.The impurity content of filtered liquid medicine is evaluated, and the adjustment necessity of the liquid medicine filtering process is judged according to the impurity content evaluation score; if adjustment necessity exists, performing linear correlation judgment on the centrifugal speed and impurity content evaluation score in the medicine liquid filtering process, and if linear correlation exists, calculating to obtain the first adjustment amount of the centrifugal speed; the filtered liquid medicine is obtained through the adjusted centrifugal speed, the biological activity is analyzed to obtain a biological activity score, and whether the biological activity of the liquid medicine meets the standard or not is judged. In addition, a double-protection mechanism is also arranged, and the filtering quality is prevented from being influenced by reverse adjustment by adjusting direction consistency judgment, for example, after the centrifugal speed is increased for the first time, if secondary adjustment needs to be reduced, automatic termination is performed, and the process stability is guaranteed.

Description

Technical Field

[0001] The present invention relates to the technical field of biopharmaceuticals, and particularly relates to a method and system for extracting and filtering liquid medicine for biopharmaceuticals. Background Art

[0002] Biological drugs refer to products used for prevention, treatment, and diagnosis, which are manufactured by using biological organisms, biological tissues, cells, body fluids, etc. by applying the research results of biology, medicine, biochemistry, etc. and comprehensively utilizing the principles and methods of disciplines such as physics, chemistry, biochemistry, biotechnology, and pharmacy. Biopharmaceuticals have high purity requirements and need to remove harmful substances or impurities through bioseparation and purification technologies without destroying the activity of the target product. However, in the current analysis and processing of the filtered liquid medicine in the biopharmaceutical process, attention is often paid to the effectiveness of two aspects: impurity content and biological activity. However, there is a lack of a dual-protection judgment for establishing adjustment between impurity control and activity retention, which may lead to adjustment contradictions during the production process. For example, reducing the centrifugation speed to retain activity may result in excessive impurities, and excessive impurities may then require an increase in speed, resulting in damage to the activity again, thereby affecting production efficiency. Summary of the Invention

[0003] The purpose of the present invention is to provide a method and system for extracting and filtering liquid medicine for biopharmaceuticals to solve at least one of the above-mentioned prior art problems.

[0004] In the first aspect, the present invention provides a method for extracting and filtering liquid medicine for biopharmaceuticals, including the following steps: Evaluate the impurity content of the filtered liquid medicine, and judge the necessity of adjustment during the liquid medicine filtration process through the impurity content evaluation score; If there is a need for adjustment, perform a linear correlation judgment on the centrifugation speed and the impurity content evaluation score during the liquid medicine filtration process. If there is a linear correlation, calculate the first centrifugation speed adjustment amount; Obtain the filtered liquid medicine using the adjusted centrifugation speed, analyze the biological activity to obtain the biological activity score, and judge whether the biological activity of the liquid medicine meets the standard; If the biological activity of the liquid medicine does not meet the standard, perform a linear correlation judgment on the centrifugation speed and the biological activity score during the liquid medicine filtration process. If there is a linear correlation, calculate the second centrifugation speed adjustment amount; Judge whether to perform a second adjustment on the centrifugation speed according to the adjustment directions of the second centrifugation speed adjustment amount and the first centrifugation speed adjustment amount.

[0005] In the second aspect, the present invention provides a system for extracting and filtering liquid medicine for biopharmaceuticals, and the system includes: Adjustment necessity analysis module: Evaluate the impurity content of the filtered liquid medicine, and judge the necessity of adjusting the liquid medicine filtration process through the impurity content evaluation score; First adjustment analysis module: If adjustment is necessary, judge the linear correlation between the centrifugal speed and the impurity content evaluation score in the liquid medicine filtration process. If there is a linear correlation, calculate the first adjustment amount of the centrifugal speed; Activity judgment module: Use the adjusted centrifugal speed to obtain the filtered liquid medicine, analyze the biological activity to obtain the biological activity score, and judge whether the biological activity of the liquid medicine meets the standard; Second adjustment analysis module: If the biological activity of the liquid medicine does not meet the standard, judge the linear correlation between the centrifugal speed and the biological activity score in the liquid medicine filtration process. If there is a linear correlation, calculate the second adjustment amount of the centrifugal speed; Adjustment direction analysis module: Judge whether to make a second adjustment to the centrifugal speed according to the adjustment directions of the second adjustment amount and the first adjustment amount of the centrifugal speed.

[0006] Advantages of the present invention: Through the closed-loop process of "impurity detection - centrifugal parameter adjustment - biological activity verification", the present invention improves the liquid medicine to meet both the impurity content standard and the biological activity requirement, increases the efficiency of the liquid medicine filtration process in biopharmaceuticals, and establishes a linear regression model of centrifugal speed with impurities and activity based on historical data to accurately calculate the adjustment amount, realizing parameter target optimization and reducing the trial-and-error cost; The present invention also sets up a double protection mechanism. Through the judgment of the adjustment direction consistency, it avoids the influence on the filtration quality caused by reverse adjustment. For example, after increasing the centrifugal speed for the first time, if the second adjustment needs to decrease, it will be automatically terminated to ensure the process stability. Description of the drawings

[0007] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0008] Figure 1 It is a flowchart of a method for extracting and filtering liquid medicine in biopharmaceuticals of the present invention; Figure 2 It is a schematic structural diagram of a system for extracting and filtering liquid medicine in biopharmaceuticals of the present invention. Detailed implementation manners

[0009] To enable those skilled in the art to better understand the solution of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the scope of protection of the present invention.

[0010] Embodiment 1: The centrifugation rate directly affects the separation efficiency of impurities in the liquid medicine and may also damage the protein structure through mechanical stress. Therefore, the centrifugation rate is the link connecting "impurity control" and "activity retention". As Figure 1 shown, a method for extracting and filtering liquid medicine for biopharmaceuticals provided by an embodiment of the present invention specifically includes steps S1 to S5, and these steps will be described in detail below: S1: Evaluate the impurity content of the filtered liquid medicine, and judge the necessity of adjusting the liquid medicine filtration process based on the impurity content evaluation score. In this embodiment, a sterile sampling valve is installed at the end of the liquid medicine filtration system for biopharmaceuticals to sample the filtered liquid medicine. Among them, the number of samples can be several, and the samples are marked with sub-numbers, time, and sampling points. Process the sampled samples. Among them, the processing process includes: if the sample is turbid, centrifuge to remove particulate impurities and place the sample in a refrigerator at 2-8°C. A preferred implementation manner of this step includes: detecting the impurity content of the liquid medicine through multiple indicators. Among them, the types of impurities in the liquid medicine include: physical impurities, chemical impurities, and biological impurities. Specifically, the physical impurity indicators include: particulate impurities and visible foreign matters; the chemical impurity indicators include: protein aggregates, DNA residues, and host cell proteins; the biological impurity indicators include: endotoxin. Furthermore, according to the types of impurities, multi-index joint detection is adopted, specifically: For the detection of physical impurity indicators: The light scattering method or the microscopic counting method is used for particle calculation; the lamp inspection method or machine vision detection is used for the inspection of visible foreign matters. For the detection of chemical impurity indicators: Size exclusion chromatography or dynamic light scattering (DLS) is used for the detection of protein aggregates; fluorescence quantitative PCR and DNA hybridization methods are used for the detection of DNA residues; enzyme-linked immunosorbent assay is used for the detection of host cell proteins. For the detection of biological impurity indicators: The limulus reagent method and the dynamic turbidity method are used for the detection of endotoxin. For the detected multi-index impurities, a fuzzy comprehensive evaluation method is used to obtain a multi-level comprehensive quantitative evaluation index. The specific process is as follows: Determine the evaluation levels, specifically: The first-level indicators of the evaluation levels are: U = {physical impurities, chemical impurities, biological impurities}; The second-level indicators of the evaluation levels are: Physical impurities U1 = {particle impurities, visible foreign matters}; Chemical impurities U2 = {protein aggregates, DNA residues, host cell proteins}; Biological impurities U3 = {endotoxin}; Set 5-level comments and assign quantitative scores: V = {V1 (excellent, 90 - 100), V2 (good, 80 - 90), V3 (qualified, 70 - 80), V4 (nearly unqualified, 60 - 70), V5 (unqualified, <60)}; According to the relationship between the detection value and the standard, calculate the membership degree of each index to each comment through the membership function; for the second-level indicators under each first-level indicator, form a matrix after calculating the membership degree; Determine the index weights by the method of expert scoring; Conduct fuzzy synthesis for each first-level indicator, that is, the weight to the membership degree vector of each comment; The membership degree matrix of the first-level indicators Is synthesized with the first-level weight W=(0.2, 0.3, 0.5) to obtain the final comprehensive evaluation vector: Select the comment level with the highest membership degree (for example, B=(0.2, 0.3, 0.4, 0.1, 0) corresponds to qualified); Use the weighted average method to calculate the comprehensive evaluation score as the impurity content evaluation score; If the impurity content evaluation score is greater than the score limit value, it indicates that the filtration process is qualified, and a signal of no adjustment necessity is generated; If the impurity content evaluation score is less than or equal to the score limit value, it indicates that the filtration process is unqualified, and a signal of high adjustment necessity is generated; The effect of evaluating the impurity content evaluation score of the filtered liquid medicine through multi-index comprehensive evaluation is that: if a single index is used, it is easy to ignore other types of impurities, and using fuzzy comprehensive evaluation quantifies the risk and reduces subjective judgment; S2: If a signal of high adjustment necessity is generated, make a linear correlation judgment on the centrifugal speed and the impurity content evaluation score in the liquid medicine filtration process. If there is a linear correlation, calculate the first adjustment amount of the centrifugal speed according to the correlation relationship; In this step, specifically, the process of making a linear correlation judgment on the centrifugal speed and the impurity content evaluation score in the liquid medicine filtration process is as follows: Based on historical data, obtain the centrifugal speed data sequence and the corresponding impurity content evaluation score sequence; Use the Pearson correlation coefficient to determine whether there is a linear correlation between the centrifugation speed and the evaluation score of impurity content. The specific process is as follows: The calculation formula for the Pearson correlation coefficient r is: ; where and respectively represent the i-th centrifugation speed and the evaluation score of impurity content, and respectively represent the mean value of the centrifugation speed and the mean value of the evaluation score of impurity content; Based on the calculated Pearson correlation coefficient, combined with hypothesis testing to judge whether the linear correlation is significant. Specifically: The null hypothesis, that is, assuming that there is no linear correlation between the centrifugation speed data sequence and the evaluation score sequence of impurity content, that is, r = 0, is the starting point of the test; The alternative hypothesis, which is opposite to the null hypothesis, that is, assuming that there is a linear correlation between the centrifugation speed data sequence and the evaluation score sequence of impurity content, that is, r ≠ 0. If there is sufficient evidence to reject the null hypothesis, it supports the alternative hypothesis, proving that there is a linear association between the data; Calculate the test statistic t. The calculation formula is: ; where n represents the number of data; Convert the Pearson correlation coefficient into a statistic that follows a t-distribution with degrees of freedom df = n - 2, which is used to measure whether the difference between r and 0 is significant; The degrees of freedom represent the number of independent variables when calculating the statistic. In correlation analysis, since two parameters (the slope and intercept of the regression line) need to be estimated, 2 is subtracted from the sample size n; Calculate the p-value through the t-distribution. According to the t-value and degrees of freedom, look up the critical value table of the t-distribution to find the corresponding probability range; Use statistical tools, such as the T.DIST.2T function in Excel and the scipy.stats.t.sf function in Python, input the t-value and degrees of freedom, and directly output the p-value; Compare the p-value with the significance level α. Among them, the significance level α is usually set to 0.05; If p ≤ α, it means that under the premise that the null hypothesis holds, the probability of observing the current data (or more extreme data) is extremely low (less than 5%). At this time, reject the null hypothesis, that is, there is a linear correlation between the centrifugation speed and the evaluation score of impurity content; Otherwise, it is judged that there is no linear correlation between the centrifugation speed and the evaluation score of impurity content; based on the non-existence of linear correlation, other parameters are analyzed; In this step, specifically, the process of calculating the centrifugation speed adjustment amount according to the correlation relationship includes: constructing a linear regression equation based on the centrifugation speed data sequence and the evaluation score sequence of impurity content, and then calculating the first centrifugation speed adjustment amount through the linear regression equation; Further, as a preferred implementation of this step, it includes: Taking the centrifugal speed as the X value and the impurity content evaluation score as the y value, performing linear regression on the centrifugal speed data sequence and the impurity content evaluation score sequence, and setting the linear regression equation as: ; Calculating the intercept a and slope b of the regression coefficient in the linear regression equation respectively; Specifically, the calculation formula for the slope b is: ; the calculation formula for the intercept a is: ; Constructing a linear regression equation based on the intercept a and slope b; Setting the target impurity content evaluation score, inputting the target impurity content evaluation score into the linear regression equation, and outputting to obtain the first adjustment target value of the centrifugal speed; Based on the current centrifugal speed and the first adjustment target value of the centrifugal speed, calculating the first adjustment amount of the centrifugal speed; Adjusting the centrifugal speed of the liquid medicine filtration process through the first adjustment amount of the centrifugal speed; In this embodiment, the effect of S2 lies in: through Pearson correlation analysis and linear regression model, analyzing the quantitative relationship between the centrifugal speed and the impurity content, and improving the adjustment efficiency; S3: Continuing to filter the liquid medicine using the adjusted centrifugal speed, analyzing the biological activity of the filtered liquid medicine to obtain a biological activity score, and judging whether the biological activity of the liquid medicine meets the standard; In this step, after the first adjustment of the centrifugal speed, sampling is carried out after filtering the liquid medicine, where the sampling point is the same as the sampling point in S1; The specific process of detecting the biological activity of the liquid medicine includes detecting two indicators: functional activity and structural integrity; First specifically, the detection process of functional activity includes: measuring the relative activity unit using a cell proliferation inhibition experiment; Selecting a cell line sensitive to the target drug and culturing it to the logarithmic growth phase in a suitable culture medium; Adjusting the cell density (such as ells / well), inoculating it into a 96-well plate, and culturing for 24 hours; Diluting the liquid medicine into gradient concentrations (such as 100 ng / mL, 10 ng / mL, 1 ng / mL), adding it to the wells, and setting 3 - 6 replicates for each group; At the same time, setting a negative control (without drug) and a positive control (standard drug); After culturing for 48 - 72 hours, adding a cell viability dye (such as CCK-8, MTT), and incubating for 2 hours; Measure the absorbance with a microplate reader (e.g., at a wavelength of 450 nm) and calculate the cell survival rate. The calculation formula for the cell survival rate is as follows: ; Plot the dose-response curve and fit to obtain the concentration at which the drug inhibits 50% of cell proliferation ( ). The calculation formula for the relative activity unit is as follows: ; Second specifically, the detection process of structural integrity includes: Detect the secondary structure by circular dichroism (CD). The process is as follows: Dilute the drug solution to 0.1 - 1 mg / mL and place it in a quartz cuvette; Scan the wavelength from 190 - 260 nm on a CD spectrometer and record the ellipticity (θ); Perform baseline correction (subtract the buffer background); Use software (such as CDPro) to fit the spectrum and calculate the percentage of each secondary structure; Perform weighted calculation on the relative activity unit and the percentage of secondary structure to obtain the bioactivity score; If the bioactivity is greater than or equal to the bioactivity limit value, the bioactivity of the filtered drug solution meets the standard; If the bioactivity is less than the bioactivity limit value, the bioactivity of the filtered drug solution does not meet the standard; S4: If the bioactivity of the drug solution does not meet the standard, perform a linear correlation judgment on the centrifugation speed and the bioactivity score during the drug solution filtration process. If there is a linear correlation, calculate the secondary adjustment amount of the centrifugation speed according to the correlation relationship; In this step, first specifically, the process of performing a linear correlation judgment on the centrifugation speed and the bioactivity during the drug solution filtration process is as follows: Based on historical data, obtain the centrifugation speed data sequence and the corresponding bioactivity score sequence; Use the Pearson correlation coefficient to judge whether there is a linear correlation between the centrifugation speed and the bioactivity score. The specific process is as follows: The calculation formula for the Pearson correlation coefficient r is as follows: ; where, and respectively represent the j-th centrifugation speed and bioactivity score, and respectively represent the mean value of the centrifugation speed and the mean value of the bioactivity score; Based on the calculated Pearson correlation coefficient, combine the hypothesis test to judge whether the linear correlation is significant. Specifically: The original hypothesis, that is, assume that there is no linear correlation between the centrifugation speed data sequence and the bioactivity score sequence, that is, r = 0, is the starting point of the test; The alternative hypothesis, which is opposite to the null hypothesis, assumes that there is a linear correlation between the centrifugation speed data sequence and the bioactivity score sequence, that is, r≠0. If there is sufficient evidence to reject the null hypothesis, the alternative hypothesis is supported, proving that there is a linear correlation between the data; Calculate the test statistic t, and the calculation formula is: ; where n represents the number of data; Convert the Pearson correlation coefficient into a statistic that follows a t-distribution with degrees of freedom df = n - 2, which is used to measure whether the difference between r and 0 is significant; The degrees of freedom represent the number of independent variables when calculating the statistic. In correlation analysis, since two parameters (the slope and intercept of the regression line) need to be estimated, 2 is subtracted from the sample size n; Calculate the p-value through the t-distribution. According to the t-value and degrees of freedom, look up the critical value table of the t-distribution to find the corresponding probability range; Use statistical tools, such as the T.DIST.2T function in Excel and the scipy.stats.t.sf function in Python, input the t-value and degrees of freedom, and directly output the p-value; Compare the p-value with the significance level α, where the significance level α is usually set to 0.05; If p ≤ α, it means that under the premise that the null hypothesis holds, the probability of observing the current data (or more extreme data) is extremely low (less than 5%). At this time, the null hypothesis is rejected, that is, there is a linear correlation between the centrifugation speed and the bioactivity score; Otherwise, it is judged that there is no linear correlation between the centrifugation speed and the bioactivity score; based on the absence of linear correlation, other parameters are analyzed; In this step, specifically, the process of calculating the centrifugation speed adjustment amount according to the correlation relationship includes: constructing a linear regression equation based on the centrifugation speed data sequence and the bioactivity score sequence, and then calculating the secondary centrifugation speed adjustment amount through the linear regression equation; Furthermore, as a preferred implementation manner of this step includes: Taking the centrifugation speed as the X value and the impurity content evaluation score as the y value, perform a linear regression on the centrifugation speed data sequence and the impurity content evaluation score sequence, and assume the linear regression equation is: ; Calculate the intercept c and slope d of the regression coefficient in the linear regression equation respectively; Specifically, the calculation formula for the slope d is: ; the calculation formula for the intercept c is: ; Construct a linear regression equation based on the intercept c and slope d; Set the target bioactivity score, input the target bioactivity score into the linear regression equation, and output the secondary adjustment target value of the centrifugation speed; Calculate the secondary adjustment amount of the centrifugal speed based on the first centrifugal speed and the target value of the secondary adjustment of the centrifugal speed. In this embodiment, the effect of S4 is as follows: After the first adjustment of the centrifugal speed, verify the effectiveness through bioactivity detection. If the activity does not meet the standard, initiate secondary adjustment analysis to form a closed-loop control of impurity control → activity verification → parameter optimization, so that the preparation of the liquid medicine meets both the impurity standard and the bioactivity requirement, thereby improving production efficiency. S5: Determine whether the adjustment directions of the secondary adjustment amount of the centrifugal speed and the first adjustment amount of the centrifugal speed are the same. If they are the same, perform a secondary adjustment on the centrifugal speed. In this step, identify the adjustment directions of the first adjustment amount of the centrifugal speed and the secondary adjustment amount of the centrifugal speed. If both the first adjustment amount of the centrifugal speed and the secondary adjustment amount of the centrifugal speed are increasing or both are decreasing, it indicates that the adjustment directions of the first adjustment amount of the centrifugal speed and the secondary adjustment amount of the centrifugal speed are consistent. Otherwise, it indicates that the adjustment directions of the first adjustment amount of the centrifugal speed and the secondary adjustment amount of the centrifugal speed are inconsistent. If they are inconsistent, adjust other parameters. If the adjustment directions of the secondary rotation rate adjustment amount and the primary rotation rate adjustment amount are consistent, readjust the rotation rate of the crusher according to the secondary rotation rate adjustment amount. In this example, the process of S5 is to determine whether the secondary adjustment amount of the centrifugal speed will affect the evaluation score of the impurity content after filtering the liquid medicine. The reason for the judgment is: When making the first adjustment of the centrifugal speed, the first adjustment amount is to make the evaluation score of the impurity content reach the target impurity evaluation score (i.e., the critical value of the impurity content standard). If the direction of the secondary adjustment amount of the centrifugal speed is different from that of the first adjustment amount of the centrifugal speed (such as the case where the first adjustment amount of the centrifugal speed increases and the secondary adjustment amount of the centrifugal speed decreases), if the centrifugal speed is continuously adjusted for the second time, it will affect the evaluation score of the impurity content and make it not meet the impurity content standard. The technical solution of this embodiment is as follows: After end sampling, detect through three major categories of physical, chemical, and biological indicators, calculate the impurity content score through fuzzy comprehensive evaluation, and judge whether to trigger adjustment; if adjustment is required, calculate the first adjustment amount of the centrifugal speed based on the Pearson correlation analysis and linear regression model of historical data, and target to improve the impurity score; after adjustment, detect the functional activity and structural integrity of the liquid medicine, and calculate the bioactivity score through weighting to verify the effectiveness of the process; if the activity does not meet the standard, analyze the correlation between the centrifugal speed and the activity again to generate a secondary adjustment amount; through the judgment of the adjustment direction consistency, decide whether to perform the secondary adjustment to avoid the risk of exceeding the impurity standard caused by reverse adjustment. Thus, through the closed-loop control of multi-dimensional impurity detection and bioactivity verification, the present invention combines a statistical model to adjust the centrifugation speed, improving the production efficiency of bio-pharmaceutical liquid filtration; covering 9 impurity indicators of physical, chemical, and biological properties in all dimensions, reducing the risk of missed detection; based on the Pearson correlation and linear regression models, achieving targeted optimization of the centrifugation speed for the first adjustment; and by judging the consistency of the adjustment directions of the first adjustment and the second adjustment, avoiding fluctuations in the impurity content of the liquid filtration caused by reverse adjustment.

[0011] Embodiment 2: Based on the above embodiment, as Figure 2 shown, a liquid extraction and filtration system for bio-pharmaceuticals provided by an embodiment of the present invention specifically includes: Adjustment necessity analysis module: Evaluate the impurity content of the filtered liquid medicine, and judge the necessity of adjustment in the liquid medicine filtration process through the impurity content evaluation score; In this embodiment, a sterile sampling valve is installed at the end of the liquid medicine filtration system for bio-pharmaceuticals to sample the filtered liquid medicine. Among them, the number of samples can be several, and the samples are marked with sub-numbers, time, and sampling points; Process the sampled samples. Among them, the processing process includes: If the sample is turbid, centrifuge to remove particulate impurities and refrigerate the sample at 2-8°C; Detect the impurity content of the liquid medicine through multiple indicators; Among them, the types of impurities in the liquid medicine include: physical impurities, chemical impurities, and biological impurities; Specifically, the physical impurity indicators include: particulate impurities and visible foreign matters; the chemical impurity indicators include: protein aggregates, DNA residues, and host cell proteins; the biological impurity indicators include: endotoxin; Furthermore, according to the type of impurities, multi-index joint detection is adopted, For the multi-index impurities obtained by detection, a fuzzy comprehensive evaluation method is used to obtain a multi-level comprehensive quantitative evaluation index, and then the impurity content evaluation score is calculated; If the impurity content evaluation score is greater than the score limit value, it means that the filtration process is qualified, and a signal of no adjustment necessity is generated; If the impurity content evaluation score is less than or equal to the score limit value, it means that the filtration process is unqualified, and a signal of high adjustment necessity is generated; First adjustment analysis module: If a signal of high adjustment necessity is generated, judge the linear correlation between the centrifugation speed and the impurity content evaluation score in the liquid medicine filtration process. If there is a linear correlation, calculate the first adjustment amount of the centrifugation speed according to the correlation relationship; In this embodiment, based on historical data, obtain the centrifugation speed data sequence and the corresponding impurity content evaluation score sequence; Use the Pearson correlation coefficient to determine whether there is a linear correlation between the centrifugation speed and the evaluation score of the impurity content; Based on the calculated Pearson correlation coefficient, combined with hypothesis testing to judge whether the linear correlation is significant, calculate the test statistic t; calculate the p-value through the t-distribution; Compare the p-value with the significance level α, where the significance level α is usually set to 0.05; If p ≤ α, it indicates that there is a linear correlation between the centrifugation speed and the evaluation score of the impurity content; Otherwise, it is judged that there is no linear correlation between the centrifugation speed and the evaluation score of the impurity content; based on the absence of linear correlation, other parameters are analyzed; Construct a linear regression equation based on the centrifugation speed data sequence and the evaluation score sequence of the impurity content; Set the target evaluation score of the impurity content, input the target evaluation score of the impurity content into the linear regression equation, and output the first adjustment target value of the centrifugation speed; Based on the current centrifugation speed and the first adjustment target value of the centrifugation speed, calculate the first adjustment amount of the centrifugation speed; Adjust the centrifugation speed of the liquid medicine filtration process through the first adjustment amount of the centrifugation speed; Activity judgment module: Use the adjusted centrifugation speed to continue filtering the liquid medicine, analyze the biological activity of the filtered liquid medicine to obtain the biological activity score, and judge whether the biological activity of the liquid medicine meets the standard; In this embodiment, after the first adjustment of the centrifugation speed, sampling is carried out after filtering the liquid medicine; The specific process of detecting the biological activity of the liquid medicine includes detecting two indicators of functional activity and structural integrity, and obtaining the relative activity unit and the percentage of secondary structure; Perform weighted calculation on the relative activity unit and the percentage of secondary structure to obtain the biological activity score; If the biological activity is greater than or equal to the biological activity limit value, the biological activity of the filtered liquid medicine meets the standard; If the biological activity is less than the biological activity limit value, the biological activity of the filtered liquid medicine does not meet the standard; Secondary adjustment analysis module: If the biological activity of the liquid medicine does not meet the standard, judge the linear correlation between the centrifugation speed and the biological activity score in the liquid medicine filtration process. If there is a linear correlation, calculate the secondary adjustment amount of the centrifugation speed according to the correlation relationship; In the embodiment, based on historical data, obtain the centrifugation speed data sequence and the corresponding biological activity score sequence; Use the Pearson correlation coefficient to determine whether there is a linear correlation between the centrifugation speed and the biological activity score, Based on the calculated Pearson correlation coefficient, combined with hypothesis testing to determine whether the linear correlation is significant, calculate the test statistic t; calculate the p-value through the t-distribution; Compare the p-value with the significance level α, where the significance level α is usually set to 0.05; If p ≤ α, it indicates that there is a linear correlation between the centrifugation speed and the bioactivity score; Otherwise, it is determined that there is no linear correlation between the centrifugation speed and the bioactivity score; based on the absence of linear correlation, other parameters are analyzed; Construct a linear regression equation based on the centrifugation speed data sequence and the bioactivity score sequence, and then calculate the quadratic adjustment amount of the centrifugation speed through the linear regression equation; Set the target bioactivity score, input the target bioactivity score into the linear regression equation, and output the quadratic adjustment target value of the centrifugation speed; Based on the initial centrifugation speed and the quadratic adjustment target value of the centrifugation speed, calculate the quadratic adjustment amount of the centrifugation speed; Adjustment direction analysis module: Determine whether the adjustment directions of the quadratic adjustment amount of the centrifugation speed and the initial adjustment amount of the centrifugation speed are the same. If they are the same, the centrifugation speed is adjusted quadraticly; In the embodiment, identify the adjustment directions of the initial adjustment amount of the centrifugation speed and the quadratic adjustment amount of the centrifugation speed; If both the initial adjustment amount of the centrifugation speed and the quadratic adjustment amount of the centrifugation speed are increasing or both are decreasing, the rotation speed of the crusher is adjusted again according to the quadratic rotation speed adjustment amount.

[0012] In the above embodiments, the descriptions of each embodiment have their own focuses. For parts not detailed or recorded in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0013] The above has described an embodiment of the present invention in detail, but the content described is only a preferred embodiment of the present invention and cannot be considered as limiting the scope of implementation of the present invention. All equivalent changes and improvements made according to the scope of the present invention application should still fall within the patent coverage scope of the present invention.

Claims

1. A method for extracting and filtering a liquid medicine for biopharmaceuticals, characterized in that, It includes the following steps: Evaluate the impurity content of the filtered liquid medicine, and judge the necessity of adjusting the liquid medicine filtration process through the impurity content evaluation score; If adjustment is necessary, judge the linear correlation between the centrifugal speed and the impurity content evaluation score in the liquid medicine filtration process. If there is a linear correlation, calculate the first centrifugal speed adjustment amount; Obtain the filtered liquid medicine using the adjusted centrifugal speed, analyze the biological activity to obtain the biological activity score, and judge whether the biological activity of the liquid medicine meets the standard; If the biological activity of the liquid medicine does not meet the standard, judge the linear correlation between the centrifugal speed and the biological activity score in the liquid medicine filtration process. If there is a linear correlation, calculate the second centrifugal speed adjustment amount; Judge whether to make a second adjustment to the centrifugal speed according to the adjustment directions of the second centrifugal speed adjustment amount and the first centrifugal speed adjustment amount.

2. The method for extracting and filtering the liquid medicine for biopharmaceuticals according to claim 1, wherein The process of judging the necessity of adjusting the liquid medicine filtration process is as follows: Detect the impurity content of the liquid medicine through multiple indicators; for the multiple - indicator impurities obtained from the detection, use the fuzzy comprehensive evaluation method to obtain the multi - level impurity content evaluation score; If the impurity content evaluation score is less than or equal to the score limit value, adjustment is necessary.

3. A method for extracting and filtering a liquid medicine for biopharmaceuticals according to claim 2, characterized in that, The multiple indicators include: particulate impurities and visible foreign matters; protein aggregates, DNA residues, host cell proteins, and endotoxins.

4. A method for extracting and filtering a liquid medicine for biopharmaceuticals according to claim 1, characterized in that, The process of judging the linear correlation between the centrifugal speed and the impurity content evaluation score in the liquid medicine filtration process is as follows: Based on historical data, obtain the centrifugal speed data sequence and the corresponding impurity content evaluation score sequence, and calculate the Pearson correlation coefficient; Based on the calculated Pearson correlation coefficient; Calculate the test statistic t, convert the Pearson correlation coefficient to follow a t - distribution; calculate the p - value through the t - distribution. The p - value is the probability of observing the current sample data result under the premise that the null hypothesis holds; Compare the p - value with the significance level α. If p ≤ α, it indicates that there is a linear correlation.

5. A method for extracting and filtering a liquid medicine for biopharmaceuticals according to claim 1, characterized in that, The process of calculating the first centrifugal speed adjustment amount is as follows: Construct a linear regression equation based on the centrifugal speed data sequence and the impurity content evaluation score sequence, input the target impurity content evaluation score into the linear regression equation, and output the first centrifugal speed adjustment target value; Based on the current centrifugal speed and the first centrifugal speed adjustment target value, calculate the first centrifugal speed adjustment amount.

6. A method for extracting and filtering a liquid medicine for biopharmaceuticals according to claim 1, characterized in that, The process of judging whether the biological activity of the liquid medicine meets the standard is as follows: The specific process of detecting the biological activity of the liquid medicine includes detecting two indicators: functional activity and structural integrity; Based on the monitoring results of the detection of the two indicators of functional activity and structural integrity, obtain the relative activity unit and the secondary structure percentage, and perform weighted calculation on the relative activity unit and the secondary structure percentage to obtain the biological activity score.

7. A method for extracting and filtering a liquid medicine for biopharmaceuticals according to claim 1, characterized in that, The process of judging the linear correlation between the centrifugal speed and the biological activity score in the liquid medicine filtration process is as follows: Based on historical data, obtain the centrifugal speed data sequence and the corresponding biological activity score sequence, and calculate the Pearson correlation coefficient; Based on the calculated Pearson correlation coefficient; Calculate the test statistic t, convert the Pearson correlation coefficient to follow a t - distribution; calculate the p - value through the t - distribution; Compare the p-value with the significance level α. If p ≤ α, it indicates the existence of a linear correlation.

8. A method for extracting and filtering a liquid medicine for biopharmaceuticals according to claim 1, characterized in that, The process of calculating the secondary adjustment amount of the centrifugation speed is as follows: Construct a linear regression equation based on the centrifugation speed data sequence and the bioactivity score sequence, input the target bioactivity score into the linear regression equation, and output the target value of the secondary adjustment of the centrifugation speed; Based on the initial centrifugation speed and the target value of the secondary adjustment of the centrifugation speed, calculate the secondary adjustment amount of the centrifugation speed.

9. A method for extracting and filtering a liquid medicine for biopharmaceuticals according to claim 1, characterized in that, The process of determining whether to perform a secondary adjustment on the centrifugation speed is as follows: Identify the adjustment directions of the initial adjustment amount of the centrifugation speed and the secondary adjustment amount of the centrifugation speed; If both the initial adjustment amount of the centrifugation speed and the secondary adjustment amount of the centrifugation speed are increasing or both are decreasing, then readjust the rotation speed of the crusher according to the secondary adjustment amount of the rotation speed.

10. A liquid extraction and filtration system for biopharmaceutical drugs, characterized in that, This system is used to execute the method described in any one of the above claims 1-9. This system includes: Adjustment necessity analysis module: Evaluate the impurity content of the filtered liquid medicine, and judge the necessity of adjusting the liquid medicine filtration process through the impurity content evaluation score; Initial adjustment analysis module: If there is a specific necessity for adjustment, judge the linear correlation between the centrifugation speed and the impurity content evaluation score in the liquid medicine filtration process. If there is a linear correlation, calculate the initial adjustment amount of the centrifugation speed; Activity judgment module: Obtain the filtered liquid medicine using the adjusted centrifugation speed, analyze the bioactivity to obtain the bioactivity score, and judge whether the bioactivity of the liquid medicine meets the standard; Secondary adjustment analysis module: If the bioactivity of the liquid medicine does not meet the standard, judge the linear correlation between the centrifugation speed and the bioactivity score in the liquid medicine filtration process. If there is a linear correlation, calculate the secondary adjustment amount of the centrifugation speed; Adjustment direction analysis module: Judge whether to perform a secondary adjustment on the centrifugation speed according to the adjustment directions of the secondary adjustment amount of the centrifugation speed and the initial adjustment amount of the centrifugation speed.

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