Glucose concentration control method based on accumulated oxygen transfer rate and application thereof

Abstract

The invention provides a glucose concentration control method based on an accumulated oxygen transfer rate and application thereof. According to the method, conventional cell culture online control parameters such as dissolved oxygen level, ventilation rate, stirring speed, reactor geometric dimension and representative historical batch data are collected, accumulated oxygen mass transfer of reactor online detection data is calculated, and the model is corrected through model operation and historical data, so that accumulated sugar consumption is accurately predicted; and further calculating to obtain the concentration of the residual glucose in the current culture solution. According to the prediction method, only conventional on-line culture parameters are utilized, additional hardware equipment does not need to be purchased, the prediction precision is relatively sensitive, the whole process of cell culture can be predicted, and the method is suitable for small-scale and large-scale cell culture and has a wide application prospect.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a glucose concentration control method based on cumulative oxygen transfer rate and an application thereof. Background Art

[0002] Glucose is the main energy source for cells and is crucial in cell culture. On the one hand, if the glucose concentration in the cell environment is too low, insufficient glucose will cause a loss of cell energy supply, which will hinder cell growth. In addition, since glucose is also the raw material for cells to synthesize some important substances, such as nucleic acids, proteins, etc., insufficient glucose may also affect the synthesis of these substances, further affecting cell growth and division. On the other hand, when there is an excess of glucose in the cell environment, the cells will undergo glycolysis in large quantities and produce too much lactic acid, resulting in acidification of the culture environment. An overly acidic environment will have an adverse effect on various physiological activities of cells, and in severe cases may even lead to cell death. In addition, excessive glucose may also induce some cells to enter a state of metabolic disorders, thereby affecting their functions.

[0003] Therefore, in order to ensure the normal growth of cells and the quality and yield of products, it is necessary to accurately control the glucose concentration during the cell culture process and keep it at an appropriate level. This requires online monitoring and the addition or adjustment of glucose based on the monitoring results to ensure a continuous and stable supply of glucose.

[0004] There have been reports on the use of Raman technology to predict sugar concentration in fermentation broth. For example, CN109342393A discloses a method for detecting glucose content in cell culture medium using Raman spectroscopy. However, this method requires expensive Raman spectrometers and a large amount of complex spectral data machine learning and modeling.

[0005] Since cell growth and glucose metabolism require oxygen consumption, the total oxygen consumption is highly correlated with the total glucose consumption. The oxygen consumed by the bioreactor can be expressed by the cumulative OTR and calculated by the DO electrode and the oxygen mass transfer coefficient KLa. Recently, there have been reports on the use of cumulative OTR and cumulative sugar consumption to establish a mathematical model to predict the glucose concentration in the culture environment of the bioreactor. For example, in the literature "On-Line Control of Glucose Concentration in High-Yielding Mammalian Cell Cultures Enabled Through Oxygen Transfer Rate Measurements", the cumulative OTR and cumulative sugar consumption data of the last three days were used to fit a linear equation to predict the real-time cumulative sugar consumption. This method can only start prediction after obtaining the OTR and cumulative sugar consumption data of the first three experimental points, and iterate the model every day, but the prediction accuracy of this method is poor. The accuracy of the method is about ±3g / L through multiple cell line experiments. It cannot meet the requirements of process robustness control. How to further improve the prediction accuracy (<1.0g / L) to meet the actual process control needs requires further improvement of the model prediction method.

[0006] In summary, for glucose monitoring during cell culture, although some manufacturers have begun to use industrial robots or automated offline equipment for automatic sampling and automatic detection and analysis of glucose concentration, this method requires a lot of hardware investment and function development costs, and causes huge offline detection costs. On the other hand, prediction through mathematical model calculation has also received widespread attention. This method has the advantages of low cost and real-time monitoring, but how to develop a high-precision prediction model is still challenging. Summary of the invention

[0007] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a glucose concentration control method based on cumulative oxygen transfer rate and its application. The method can accurately predict the current sugar concentration and provide a basis for feedback of sugar supplementation; reduce the number of sampling and testing times of the product line, save operating steps and reduce the consumption of consumables and equipment; achieve stable sugar concentration in the cell culture environment, improve the consistency, continuity and robustness of the production process, and stabilize product quality.

[0008] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0009] In a first aspect, the present invention provides a glucose concentration control method based on cumulative oxygen transfer rate, the glucose concentration control method comprising:

[0010] S1. Collect a batch of historical process data of the same cell line and calculate the daily reference Gluc / OTR coefficient; the daily reference Gluc / OTR coefficient refers to the ratio of the difference between the cumulative Gluc and the cumulative Lac in the historical reference batch from day i to day i+1 to the corresponding cumulative OTR. ref_day(i) ;

[0011] S2. Collect online process data of cell culture, calculate cumulative OTR, and multiply the cumulative OTR by the correction factor fday(i) Obtain the corrected cumulative OTR;

[0012] S3. The daily reference Gluc / OTR coefficient is multiplied by the corrected cumulative OTR to obtain the difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment; the cumulative [Gluc] of the predicted batch at the current moment is the cumulative sugar consumption of the predicted batch from the i-th day to the i+t-th day; the cumulative [Lac] of the predicted batch at the current moment is the cumulative lactic acid consumption of the predicted batch from the i-th day to the i+t-th day;

[0013] The difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment is calculated according to Formula 12 and Formula 11, and the current moment is based on day i;

[0014] Formula 12:

[0015] (Accumulated [Gluc] target_day(i+t) – Accumulation [Gluc] target_day(i) )-(cumulative [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) )

[0016] =(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'×Ratio ref_day(i)

[0017] Among them, the accumulation of [Gluc] target_day(i+t) Indicates the instantaneous glucose concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Gluc] target_day(i) It indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch integrated over time and volume;

[0018] Accumulation [Lac] target_day(i+t) Indicates the instantaneous lactate concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Lac] target_day(i) It indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch is integrated over time and volume;

[0019] Accumulation [Gluc]target_day(i+t) -Accumulation [Gluc] target_day(i) It indicates that the instantaneous glucose concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative sugar consumption from the i-th day to the i+t-th day;

[0020] Accumulation [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) It indicates that the instantaneous lactic acid concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative lactic acid consumption from the i-th day to the i+t-th day;

[0021] (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR;

[0022] The current cumulative amount of the predicted batch [Lac] is calculated according to formula 11;

[0023] Formula 11:

[0024]

[0025] Lac ref_day(i+1) -Lac ref_day(i) It represents the change of lactic acid concentration from day i to day i+1 of the historical reference batch;

[0026] Time ref_day(i+1) -Time ref_day(i) It represents the time change of lactic acid concentration from day i to day i+1 of the historical reference batch, in days;

[0027] t represents the time span from the sampling and testing time of the batch to be predicted on the i-th day to the current time, in days;

[0028] S4. Calculate the current sugar concentration of the predicted batch using the current accumulated [Gluc] of the predicted batch. If the current sugar concentration of the predicted batch is lower than the target sugar concentration, add glucose until the concentration reaches the set value;

[0029] In a specific implementation, the accuracy is adjusted according to actual needs, such as a set value of ±0.1 g / L.

[0030] The predicted current sugar concentration of the batch is calculated according to formula 13;

[0031] Formula 13:

[0032]

[0033] In the formula, Gluc i+t Indicates the current sugar concentration;

[0034] V represents the system culture volume;

[0035] [Gluc] i represents the sugar concentration of the batch to be predicted when sampling and testing on the i-th day;

[0036] It represents the total amount of sugar supplement from day i to day i+t of the batch to be predicted;

[0037] It represents the total sugar loss of the batch to be predicted from the i-th day to the i+t-th day manually sampled.

[0038] In the present invention, the daily reference Gluc / OTR coefficient is continuously iterated every day and has different values.

[0039] In the present invention, by adding the sugar consumption and oxygen consumption ratio of historical batch reference as a basis, the ratio is further combined with the actual culture data of the current batch to be predicted for iterative correction. After monitoring the oxygen consumption of the current batch to be predicted, its sugar consumption level can be accurately calculated, and the remaining Gluc concentration can be further accurately predicted. After obtaining the current sugar concentration, the controller can be guided to perform automatic sugar replenishment operation to maintain a stable Gluc concentration, so that the entire cell culture process can be accurately predicted and controlled for sugar replenishment.

[0040] Preferably, in S1, the daily reference Gluc / OTR coefficient is calculated according to Formula 9;

[0041] Formula 9:

[0042]

[0043] Among them, the accumulation of [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i+1) represents the instantaneous glucose concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume;

[0044] Accumulation [Lac] ref_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the historical reference batch, integrated over time and volume; cumulative [Lac] ref_day(i+1) represents the instantaneous lactate concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume;

[0045] Accumulation [Gluc] ref_day(i+1) -Accumulation [Gluc] ref_day(i)represents the instantaneous glucose concentration from day i to day i+1 of the historical reference batch integrated over time and volume;

[0046] Accumulation [Lac] ref_day(i+1) -Accumulation [Lac] ref_day(i) It represents the instantaneous lactic acid concentration from day i to day i+1 of the historical reference batch integrated over time and volume;

[0047] Cumulative OTR ref_day(i+1) It represents the integration of instantaneous OTR over time and volume from day 0 to day i+1 of the historical reference batch;

[0048] Cumulative OTR ref_day(i) It represents the integration of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch.

[0049] Preferably, in S2, the cumulative OTR is the integral value of the instantaneous OUR in the time period from t0 to t, and the cumulative OTR is calculated according to Formula 6;

[0050]

[0051] Wherein, V represents the culture volume of the system.

[0052] Preferably, the instantaneous OUR is calculated according to Formula 5;

[0053] OTR=OUR=K L a×(DO * -DO) Formula 5

[0054] Where OUR represents the instantaneous oxygen transfer rate, K L a represents the gas mass transfer coefficient, DO* represents the saturated dissolved oxygen level, and DO represents the dissolved oxygen level.

[0055] Preferably, the DO is calculated according to Formula 4;

[0056]

[0057] Among them, v O2 is the oxygen flow rate, v air is the air flow rate.

[0058] Preferably, the K L a is calculated according to formula 3;

[0059]

[0060] Among them, k, α, β are constants of the empirical formula, Ws represents the apparent linear velocity, and P / V represents the output power per unit volume.

[0061] Preferably, the Ws Calculate according to formula 2;

[0062]

[0063] Among them, v O2 is the oxygen flow rate, v air is the air flow rate and Di is the reactor diameter.

[0064] Preferably, the P / V is calculated according to Formula 1;

[0065]

[0066] Among them, N p is the reactor stirring power number, Di is the reactor diameter, and N is the stirring speed.

[0067] Preferably, in S2, the correction coefficient f day(i) It represents the deviation correction coefficient between the real-time Gluc / OTR coefficient of the batch to be predicted and the Gluc / OTR coefficient of the reference batch.

[0068] In the present invention, by adding the cumulative OTR correction coefficient f day(i) Reduce the system error between different reactors to adapt to the conversion between different reactors. The significance of applying this correction factor is to eliminate the errors between different equipment and different culture batches. For example, due to the accuracy of the equipment MFC (gas mass flow meter), the detection deviation of the dissolved oxygen electrode, the sensitivity and other differences, the actual consumption of glucose per unit mass in the reactor may have a large difference in the corresponding oxygen consumption. After applying this coefficient, such errors can be greatly eliminated.

[0069] Preferably, the correction factor f day(i) Calculated according to formula 7;

[0070] Formula 7:

[0071]

[0072] Wherein, i is greater than j, j is selected from a positive integer less than 5; the number of days before and before the i-th batch of the Target are known.

[0073] Accumulation [Gluc] target_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch, integrating time and volume, and accumulating [Gluc] target_day(i-j) It indicates the instantaneous glucose concentration from day 0 to day ij of the predicted batch integrated over time and volume;

[0074] Accumulation [Lac] target_day(i)Indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch, integrated over time and volume, cumulative [Lac] target_day(i-j) It indicates the instantaneous lactate concentration from day 0 to day ij of the predicted batch integrated over time and volume;

[0075] Cumulative OTR target_day(i) It represents the integral of instantaneous OTR over time and volume from day 0 to day i of the batch to be predicted;

[0076] Cumulative OTR target_day(i-j) It represents the integral of instantaneous OTR over time and volume from day 0 to day ij of the batch to be predicted;

[0077] Accumulation [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day 0 to day ij of the historical reference batch integrated over time and volume;

[0078] Accumulation [Lac] ref_day(i) Indicates the instantaneous lactic acid concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Lac] ref_day(i-j) represents the instantaneous lactate concentration from day 0 to day ij of the historical reference batch integrated over time and volume;

[0079] Accumulation [Gluc] ref_day(i) -Accumulation [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day ij to day i of the historical reference batch integrated over time and volume;

[0080] Accumulation [Lac] ref_day(i) -Accumulation [Lac] ref_day(i-j) It represents the instantaneous lactic acid concentration from day ij to day i of the historical reference batch integrated over time and volume;

[0081] Cumulative OTR ref_day(i) represents the integral of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch;

[0082] Cumulative OTR ref_day(i-j) represents the integration of instantaneous OTR over time and volume from day 0 to day ij of the historical reference batch.

[0083] In the calculation scheme of the present invention, since the production of lactic acid in anaerobic respiration consumes glucose but not oxygen, the present invention deducts the influence of the accumulated amount of lactic acid and only associates the consumption of oxygen and glucose in aerobic respiration, which is beneficial to improve the prediction accuracy.

[0084] Preferably, the corrected cumulative OTR is calculated according to Formula 8;

[0085] Formula 8:

[0086] (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'

[0087] =(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )×f day (i)

[0088] Among them, (cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR.

[0089] The present invention collects conventional cell culture online control parameters (such as dissolved oxygen level, ventilation rate, and stirring speed, etc.), reactor geometry and representative historical batch data, etc., calculates the cumulative oxygen mass transfer of the reactor online detection data, and corrects the model through model operation and historical data, so as to accurately predict the cumulative sugar consumption, and further calculate the residual glucose concentration in the current culture solution. This prediction method only uses conventional online culture parameters without the need to purchase additional hardware equipment, has a more sensitive prediction accuracy, and the entire cell culture process is predictable. It is suitable for small-scale and large-scale cell culture and has broad application prospects.

[0090] In a second aspect, the present invention provides a model for online monitoring of glucose concentration in a cell culture process, the model comprising:

[0091] The historical process data collection unit is used to collect a batch of historical process data of the same cell line and calculate the daily reference Gluc / OTR coefficient; the daily reference Gluc / OTR coefficient refers to the ratio of the difference between the cumulative Gluc and the cumulative Lac in the historical reference batch from the i-th day to the i+1-th day to the corresponding cumulative OTR. ref_day(i) ;

[0092] The online process data collection unit is used to collect online process data of cell culture and calculate the cumulative OTR. The cumulative OTR is multiplied by the correction factor fday(i ) Obtain the corrected cumulative OTR;

[0093] The cumulative [Gluc] calculation unit of the predicted batch at the current moment, wherein the daily reference Gluc / OTR coefficient is multiplied by the corrected cumulative OTR to obtain the difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment; the cumulative [Gluc] of the predicted batch at the current moment is the cumulative sugar consumption of the predicted batch from the i-th day to the i+t-th day; the cumulative [Lac] of the predicted batch at the current moment is the cumulative lactic acid consumption of the predicted batch from the i-th day to the i+t-th day;

[0094] The difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment is calculated according to Formula 12 and Formula 11, and the current moment is based on day i;

[0095] Formula 12:

[0096] (Accumulated [Gluc] target_day(i+t) – Accumulation [Gluc] target_day(i) )-(cumulative [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) )

[0097] =(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'×Ratio ref_day(i)

[0098] Among them, the accumulation of [Gluc] target_day(i+t) Indicates the instantaneous glucose concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Gluc] target_day(i) It indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch integrated over time and volume;

[0099] Accumulation [Lac] target_day(i+t) Indicates the instantaneous lactate concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Lac] target_day(i) It indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch is integrated over time and volume;

[0100] Accumulation [Gluc] target_day(i+t) -Accumulation [Gluc] target_day(i) It indicates that the instantaneous glucose concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative sugar consumption from the i-th day to the i+t-th day;

[0101] Accumulation [Lac] target_day(i+t) -Accumulation [Lac] target_day(i)It indicates that the instantaneous lactic acid concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative lactic acid consumption from the i-th day to the i+t-th day;

[0102] (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR;

[0103] The current cumulative amount of the predicted batch [Lac] is calculated according to formula 11;

[0104] Formula 11:

[0105]

[0106] Lac ref_day(i+1) -Lac ref_day(i) It represents the change of lactic acid concentration from day i to day i+1 of the historical reference batch;

[0107] Time ref_day(i+1) -Time ref_day(i) It represents the time change of lactic acid concentration from day i to day i+1 of the historical reference batch, in days;

[0108] t represents the time span from the sampling and testing time of the batch to be predicted on the i-th day to the current time, in days;

[0109] a glucose concentration prediction calculation unit, which calculates the current sugar concentration of the predicted batch by using the current accumulated [Gluc] of the predicted batch, and if the current sugar concentration of the predicted batch is lower than the target sugar concentration, adds glucose until the concentration reaches the set value;

[0110] The predicted current sugar concentration of the batch is calculated according to formula 13;

[0111] Formula 13:

[0112]

[0113] In the formula, Gluc i+t Indicates the current sugar concentration;

[0114] V represents the system culture volume;

[0115] [Gluc] i represents the sugar concentration of the batch to be predicted when sampling and testing on the i-th day;

[0116] It represents the total amount of sugar supplement from day i to day i+t of the batch to be predicted;

[0117] It represents the total sugar loss of the batch to be predicted from the i-th day to the i+t-th day manually sampled.

[0118] Preferably, in the historical process data collection unit, the daily reference Gluc / OTR coefficient is calculated according to Formula 9;

[0119] Formula 9:

[0120]

[0121] Among them, the accumulation of [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i+1) represents the instantaneous glucose concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume;

[0122] Accumulation [Lac] ref_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the historical reference batch, integrated over time and volume; cumulative [Lac] ref_day(i+1) represents the instantaneous lactate concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume;

[0123] Accumulation [Gluc] ref_day(i+1) -Accumulation [Gluc] ref_day(i) represents the instantaneous glucose concentration from day i to day i+1 of the historical reference batch integrated over time and volume;

[0124] Accumulation [Lac] ref_day(i+1) -Accumulation [Lac] ref_day(i) It represents the instantaneous lactic acid concentration from day i to day i+1 of the historical reference batch integrated over time and volume;

[0125] Cumulative OTR ref_day(i+1) It represents the integration of instantaneous OTR over time and volume from day 0 to day i+1 of the historical reference batch;

[0126] Cumulative OTR ref_day(i) It represents the integration of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch.

[0127] Preferably, in the online process data collection unit, the cumulative OTR is the integral value of the instantaneous OUR in the time period from t0 to t, and the cumulative OTR is calculated according to Formula 6;

[0128]

[0129] Wherein, V represents the culture volume of the system.

[0130] Preferably, the instantaneous OUR is calculated according to Formula 5;

[0131] OTR=OUR=K L a×(DO * -DO) Formula 5

[0132] Where OUR represents the instantaneous oxygen transfer rate, K L a represents the gas mass transfer coefficient, DO* represents the saturated dissolved oxygen level, and DO represents the dissolved oxygen level.

[0133] Preferably, the DO is calculated according to Formula 4;

[0134]

[0135] Among them, v O2 is the oxygen flow rate, v air is the air flow rate.

[0136] Preferably, the K L a is calculated according to formula 3;

[0137]

[0138] Among them, k, α, β are constants of empirical formula, W s represents the apparent linear velocity, and P / V represents the output power per unit volume.

[0139] Preferably, the W s Calculate according to formula 2;

[0140]

[0141] Among them, v O2 is the oxygen flow rate, v air is the air flow rate and Di is the reactor diameter.

[0142] Preferably, the P / V is calculated according to Formula 1;

[0143]

[0144] Among them, N p is the reactor stirring power number, Di is the reactor diameter, and N is the stirring speed.

[0145] Preferably, in the online process data collection unit, the correction coefficient f day(i) It represents the deviation correction coefficient between the real-time Gluc / OTR coefficient of the batch to be predicted and the Gluc / OTR coefficient of the reference batch.

[0146] Preferably, the correction factor f day(i)Calculated according to formula 7;

[0147] Formula 7:

[0148]

[0149] Wherein, i is greater than j, and j is selected from a positive integer less than 5;

[0150] Accumulation [Gluc] target_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch, integrating time and volume, and accumulating [Gluc] target_day(i-j) It indicates the instantaneous glucose concentration from day 0 to day ij of the predicted batch integrated over time and volume;

[0151] Accumulation [Lac] target_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch, integrated over time and volume, cumulative [Lac] target_day(i-j) It indicates the instantaneous lactate concentration from day 0 to day ij of the predicted batch integrated over time and volume;

[0152] Cumulative OTR target_day(i) It represents the integral of instantaneous OTR over time and volume from day 0 to day i of the batch to be predicted;

[0153] Cumulative OTR target_day(i-j) It represents the integral of instantaneous OTR over time and volume from day 0 to day ij of the batch to be predicted;

[0154] Accumulation [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day 0 to day ij of the historical reference batch integrated over time and volume;

[0155] Accumulation [Lac] ref_day(i) Indicates the instantaneous lactic acid concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Lac] ref_day(i-j) represents the instantaneous lactate concentration from day 0 to day ij of the historical reference batch integrated over time and volume;

[0156] Accumulation [Gluc] ref_day(i) -Accumulation [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day ij to day i of the historical reference batch integrated over time and volume;

[0157] Accumulation [Lac] ref_day(i)-Accumulation [Lac] ref_day(i-j) It represents the instantaneous lactic acid concentration from day ij to day i of the historical reference batch integrated over time and volume;

[0158] Cumulative OTR ref_day(i) represents the integral of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch;

[0159] Cumulative OTR ref_day(i-j) represents the integration of instantaneous OTR over time and volume from day 0 to day ij of the historical reference batch.

[0160] Preferably, the corrected cumulative OTR is calculated according to Formula 8;

[0161] Formula 8:

[0162] (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'

[0163] =(Cumulative OTR target_day(i+t) – Cumulative OTR targetday(i) )×f day(i)

[0164] Among them, (cumulative OTR target_day(i+t) -Cumulative OTRt arget_day(i) )' represents the real-time corrected cumulative OTR.

[0165] In the present invention, the daily reference Gluc / OTR coefficient refers to the ratio of the cumulative Gluc consumption of the historical reference batch from day i to day i+1 excluding Lac to the corresponding cumulative OTR. ref_day(i) , that is, the slope Ratio generated by linear fitting of the cumulative Gluc consumption excluding Lac of the historical reference batch from day i to day i+1 and the corresponding cumulative OTR ref_day(i) ; Research has found that this ratio remains constant over a period of time, which is an important theoretical basis for the present invention.

[0166] In a third aspect, the present invention provides a computer-readable storage medium having a computer program / instruction stored thereon, which, when executed by a processor, implements the steps in the method for controlling glucose concentration based on cumulative oxygen transfer rate described in the first aspect.

[0167] In a fourth aspect, the present invention provides a computer device comprising a memory and a processor, wherein the memory stores a computer program / instructions, and when the computer program / instructions are executed by the processor, the steps in the glucose concentration control method based on cumulative oxygen transfer rate described in the first aspect are implemented.

[0168] The present invention also provides a method for controlling the glucose concentration in a cell culture process, the method comprising: using the glucose concentration control method based on cumulative oxygen transfer rate described in the first aspect and / or the model for online monitoring of glucose concentration in a cell culture process described in the second aspect, monitoring the glucose concentration in the cell culture process, adding glucose according to the predicted value calculated during the monitoring process, and controlling the glucose concentration.

[0169] The present invention also provides the application of the glucose concentration control method based on cumulative oxygen transfer rate described in the first aspect and / or the model for online monitoring of glucose concentration in cell culture process described in the second aspect in monitoring and controlling glucose concentration.

[0170] The present invention also provides a method for culturing cells, the method comprising: using the glucose concentration control method based on cumulative oxygen transfer rate described in the first aspect and / or the model for online monitoring of glucose concentration in the cell culture process described in the second aspect during the cell culture process, monitoring the glucose concentration in the cell culture process, and controlling the glucose concentration by adding glucose based on the calculation results.

[0171] Compared with the prior art, the present invention has the following beneficial effects:

[0172] (1) The present invention collects conventional cell culture online control parameters (such as dissolved oxygen level, ventilation rate, and stirring speed, etc.), reactor geometry and representative historical batch data, etc., calculates the cumulative oxygen mass transfer of the reactor online detection data, and adds the cumulative OTR correction coefficient to deduct the influence of the cumulative amount of lactic acid, which is conducive to improving the prediction accuracy; by adding historical batch reference data, the entire cell culture process can be accurately predicted, thereby accurately predicting the cumulative sugar consumption, and further calculating the residual glucose concentration in the current culture solution. The method can accurately predict the current sugar concentration and provide a basis for feedback sugar supplementation.

[0173] (2) The prediction method of the present invention only utilizes conventional online culture parameters without the need to purchase additional hardware equipment. It has a relatively sensitive prediction accuracy and can predict the entire cell culture process. It is suitable for small-scale and large-scale cell culture and has broad application prospects.

[0174] (3) The prediction method of the present invention can reduce the number of sampling and testing times of the product line in the experiment, save operating steps and reduce the consumption of consumables and equipment.

[0175] (4) Achieve stable sugar concentration in the cell culture environment, improve the consistency, continuity and robustness of the production process, and stabilize product quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0176] Figure 1 This is a flow chart of the application of the model of the present invention.

[0177] Figure 2 Flowchart for cumulative OTR calculation.

[0178] Figure 3 Diagram of the glucose monitoring interface for process control software.

[0179] Figure 4 The results of the reference scheme are reproduced, where Figure A is the prediction result and Figure B is the prediction deviation.

[0180] Figure 5 This is a diagram of the prediction results of the model of the present invention, wherein Figure A is the prediction result and Figure B is the prediction deviation.

[0181] Figure 6 This is a graph showing the prediction results of the model of the present invention at a 250 L culture scale, wherein Figure A is the prediction result and Figure B is the prediction deviation. DETAILED DESCRIPTION

[0182] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0183] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0184] The explanations of abbreviations in the present invention are shown in Table 1.

[0185] Table 1

[0186]

[0187]

[0188] Example 1

[0189] This example constructs a model for online monitoring of glucose concentration during cell culture. The model flow chart is as follows Figure 1 shown.

[0190] 1. Collect historical process data for a batch of the same cell line and calculate the daily reference Gluc / OTR coefficient.

[0191] 1.1 Materials and methods

[0192] The cell culture experiments corresponding to the historical process data are shown below:

[0193] The cell culture test used a CHO-K1 cell line that produces monoclonal antibodies. The culture conditions of this cell line in the seed stage shaker (Kuhner) were 36.5°C, 110rpm, 6% CO2 concentration, Gibco's CD CHO and Cytiva's Hyclone Actipro medium. The production culture stage used a 15L reactor with an initial culture volume of 8L, a culture temperature of 36.5°C, a pH setting of 6.95±0.25, a dissolved oxygen saturation of 40%, and an initial inoculation density of 0.4×10 6 cells / mL, and the feed medium was Cytiva's Hyclone Cell boost 7a / 7b (15% / 1.5%, 3% Cell boost 7a and 0.3% Cell boost 7b were added on days 3, 5, 7, 9, and 11, respectively). Sugar supplementation strategy: no sugar supplementation on days 0-4, sugar supplementation to 25 g / L on day 5, and no sugar supplementation on days 6-14.

[0194] 1.2 Offline detection and data collection

[0195] Samples were taken daily and the viable cell density was measured using Beckman's Vi-Cell XR, and the glucose, lactate, and target protein concentrations were measured using Roche's Cedex Bio analyzer.

[0196] 2. Collect online process data of cell culture and perform modeling, and obtain the glucose concentration control strategy based on the model.

[0197] 2.1 The cell culture experiment corresponding to the online process data refers to 1.1, the difference is that the sugar supplementation strategy is calculated based on the subsequent model.

[0198] 2.2 Modeling

[0199] 2.2.1 Collect real-time online data and calculate the cumulative OTR. Figure 2 The formula is used for calculation, Figure 2 Flowchart for cumulative OTR calculation.

[0200] pass Figure 2 P / V (power output per unit volume) is calculated by formula (1), where Np in formula (1) is the reactor stirring power number (constant), Di is the reactor diameter (constant), and N is the stirring speed (online monitoring parameter).

[0201] pass Figure 2 Ws (apparent linear velocity) is obtained from equation (2), where v O2 is the oxygen flow rate (online monitoring parameter), v airis the air flow rate (online monitoring parameter), Di is the reactor diameter (constant).

[0202] Will Figure 2 Substitute (1) and (2) into (3) to obtain K L a, where k, α, and β are constants of the empirical formula.

[0203] pass Figure 2 Formula (4) is used to obtain DO* and saturated dissolved oxygen level, where v O2 is the oxygen flow rate (online monitoring parameter), v air is the air flow rate (online monitoring parameter).

[0204] Will Figure 2 Substituting (3) and (4) into (5) can obtain the instantaneous OTR level, where DO is the dissolved oxygen level (online monitoring parameter).

[0205] pass Figure 2 The cumulative OUR from t0 to t can be obtained by integrating formula (6), where V is the culture volume of the system (online monitoring parameter or calculated value).

[0206] 2.2.2 Calculation of cumulative OTR correction coefficient

[0207] Usually, there may be slight differences in the assembly of different reactor accessories, and the MFG accuracy of different controllers may lead to slight deviations in the OTR calculation value in the reaction system. Therefore, the OTR of the target prediction reactor needs to be corrected to obtain an OTR with a comparable level as the reference controller. In this example, the data of the last 3 days are used to iteratively calculate the correction coefficient.

[0208] It indicates the offline monitoring data of the current prediction reactor within 3 days before the current prediction time.

[0209] Accumulation (Gluc-Lac) ref_da(i) -Accumulation (Gluc-Lac) ref_dat(i-3) This represents the reference data for the culture days equivalent to the predicted reactor, and so on. The same is true for the cumulative OTR.

[0210] Note 1: If the current day is the third day of cultivation, then the data will be the data for the previous two days, and so on.

[0211] Note 2: The concentrations of glucose and lactic acid in this formula are mass concentrations, i.e. “g / L”.

[0212] The correction factor f day(i) Calculated according to formula 7;

[0213] Formula 7:

[0214]

[0215] Wherein, i is greater than j, and j is selected from a positive integer less than 5;

[0216] Accumulation [Gluc] target_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch, integrating time and volume, and accumulating [Gluc] target_day(i-j) It indicates the instantaneous glucose concentration from day 0 to day ij of the predicted batch integrated over time and volume;

[0217] Accumulation [Lac] target_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch, integrated over time and volume, cumulative [Lac] target_day(i-j) It indicates the instantaneous lactate concentration from day 0 to day ij of the predicted batch integrated over time and volume;

[0218] Cumulative OTR target_day(i) It represents the integral of instantaneous OTR over time and volume from day 0 to day i of the batch to be predicted;

[0219] Cumulative OTR target_day(i-j) It represents the integral of instantaneous OTR over time and volume from day 0 to day ij of the batch to be predicted;

[0220] Accumulation [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day 0 to day ij of the historical reference batch integrated over time and volume;

[0221] Accumulation [Lac] ref_day(i) Indicates the instantaneous lactic acid concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Lac] ref_day(i-j) represents the instantaneous lactate concentration from day 0 to day ij of the historical reference batch integrated over time and volume;

[0222] Accumulation [Gluc] ref_day(i) -Accumulation [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day ij to day i of the historical reference batch integrated over time and volume;

[0223] Accumulation [Lac] ref_day(i) -Accumulation [Lac] ref_day(i-j) It represents the instantaneous lactic acid concentration from day ij to day i of the historical reference batch integrated over time and volume;

[0224] Cumulative OTR ref_day(i) represents the integral of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch;

[0225] Cumulative OTR ref_day(i-j) represents the integration of instantaneous OTR over time and volume from day 0 to day ij of the historical reference batch.

[0226] 2.2.3 Calculation of Corrected Cumulative OTR

[0227] The corrected cumulative OTR is calculated according to Formula 8;

[0228] Formula 8:

[0229] (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'

[0230] =(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )×f day(i)

[0231] Among them, (cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR.

[0232] 2.2.4 Calculate the daily reference Gluc / OTR coefficient.

[0233] Since glucose is directly converted into lactate without consuming oxygen, deducting lactate will help improve the accuracy and applicability of the model.

[0234] A certain amount of (sugar-lactic acid) is consumed per unit OTR, which can be obtained by referring to historical process data. Note that when the cell growth and metabolic state is relatively stable, the reference batch and the predicted batch do not necessarily need a strict correspondence in the number of culture days. For example, the proportion of the reference batch on day 10 can also be used to predict the predicted batch on day 9 or 11.

[0235] The daily reference Gluc / OTR coefficient is calculated according to Formula 9;

[0236] Formula 9:

[0237]

[0238] Among them, the accumulation of [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i+1)represents the instantaneous glucose concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume;

[0239] Accumulation [Lac] ref_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the historical reference batch, integrated over time and volume; cumulative [Lac] ref_day(i+1) represents the instantaneous lactate concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume;

[0240] Accumulation [Gluc] ref_day(i+1) -Accumulation [Gluc] ref_day(i) represents the instantaneous glucose concentration from day i to day i+1 of the historical reference batch integrated over time and volume;

[0241] Accumulation [Lac] ref_day(i+1) -Accumulation [Lac] ref_day(i) It represents the instantaneous lactic acid concentration from day i to day i+1 of the historical reference batch integrated over time and volume;

[0242] Cumulative OTR ref_day(i+1) It represents the integration of instantaneous OTR over time and volume from day 0 to day i+1 of the historical reference batch;

[0243] Cumulative OTR ref_day(i) It represents the integration of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch.

[0244] 2.2.5 Calculate the cumulative [Gluc] of the predicted batch at the current moment starting from day i, excluding the cumulative [Lac], that is, calculate the difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment, according to Formula 12.

[0245] Formula 12:

[0246] (Accumulated [Gluc] targetday(i+t) -Accumulation [Gluc] targetday(i) )-(cumulative [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) )

[0247] =(Cumulative OTR targetday(i+t) -Cumulative OTR target_day(i) )'×Ratio ref_day(i)

[0248] Among them, the accumulation of [Gluc] target_day(i+t) Indicates the instantaneous glucose concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Gluc] target_day(i)It indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch integrated over time and volume;

[0249] Accumulation [Lac] target_day(i+t) Indicates the instantaneous lactate concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Lac] target_day(i) It indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch is integrated over time and volume;

[0250] Accumulation [Gluc] target_day(i+t) -Accumulation [Gluc] target_day( i ) It indicates that the instantaneous glucose concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative sugar consumption from the i-th day to the i+t-th day;

[0251] Accumulation [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) It indicates that the instantaneous lactic acid concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative lactic acid consumption from the i-th day to the i+t-th day;

[0252] (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR;

[0253] Ratio ref_day(i) represents the daily reference Gluc / OTR coefficient.

[0254] 2.2.6 Estimate the cumulative lactic acid consumption of the current batch according to formula 11.

[0255] Formula 11:

[0256]

[0257] Lac ref_day(i+1) -Lac ref_day(i) It represents the change of lactic acid concentration from day i to day i+1 of the historical reference batch;

[0258] Time ref_day(i+1) -Time ref_day(i) It represents the time change of lactic acid concentration from day i to day i+1 of the historical reference batch, in days;

[0259] t represents the time span from the sampling and testing time of the batch to be predicted on the i-th day to the current time, in days;

[0260] 2.2.7 Calculate the cumulative sugar consumption of the current batch

[0261] Formula 10:

[0262] (Accumulated [Gluc] target_day(i+t) -Accumulation [Gluc] target_day(i) )=(cumulative [Lac] target_day(i+t) –Accumulation [Lac] target_day(i) )+(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'×Ratio ref_day(i)

[0263] Among them, the accumulation of [Gluc] target_day(i+t) -Accumulation [Gluc] target_day(i) Indicates the glucose consumption of the batch to be predicted from day i to day i+t;

[0264] Accumulation [Gluc] target_day(i+t) Indicates the integration of time and volume of the instantaneous (glucose concentration) from day 0 to day i+t of the batch to be predicted;

[0265] Accumulation [Gluc] target_day(i) Indicates the integration of time and volume of the instantaneous (glucose concentration) from day 0 to day i of the batch to be predicted;

[0266] Accumulation [Lac] target_day(i+t) Indicates the instantaneous (lactate sugar concentration) from day 0 to day i+t of the batch to be predicted, integrating the time and volume;

[0267] Accumulation [Lac] target_day(i) It represents the instantaneous (lactate sugar concentration) from day 0 to day i of the batch to be predicted, integrating time and volume.

[0268] 2.2.8 Calculate the current sugar concentration in the reactor

[0269] The current sugar concentration of the predicted batch is calculated using the accumulated [Gluc] of the predicted batch at the current moment.

[0270] The predicted current sugar concentration of the batch is calculated according to formula 13;

[0271] Formula 13:

[0272]

[0273] In the formula, Gluc i+t Indicates the current sugar concentration;

[0274] V represents the system culture volume;

[0275] [Gluc] irepresents the sugar concentration of the batch to be predicted when sampling and testing on the i-th day;

[0276] It represents the total amount of sugar supplement from day i to day i+t of the batch to be predicted;

[0277] It represents the total sugar loss of the batch to be predicted from the i-th day to the i+t-th day manually sampled.

[0278] 3. Online glucose monitoring

[0279] By developing process control software independently, we can obtain the data needed to build the above model through IOT technology, and implement feedback monitoring results for further feedback control. The software interface is as follows Figure 3 shown.

[0280] 4. Modeling results

[0281] Figure 4 These are the results of reproducing the reference scheme, where Figure A is the prediction result and Figure B is the prediction deviation. In the reproduction scheme, no prediction could be made in the first three days, and the maximum prediction error during the culture period was about 3 g / L.

[0282] Example 2

[0283] This example experimentally verifies the model described in Example 1 and analyzes the prediction effect of the constructed prediction model.

[0284] 1. The reactor culture process is as in Example 1.

[0285] 2. Open the process control software, such as Figure 3 , set basic parameters (initial culture volume, culture batch number, target sugar concentration, feed type, feed sugar concentration, historical batch data (cumulative OTR, lactate, and cumulative (sugar-lactic acid) information), turn on real-time monitoring of glucose concentration, and turn on online parameter acquisition, i.e., cumulative OTR calculation module.

[0286] 3. According to the process, timely input the feeding amount and time, sampling amount and time.

[0287] 4. If there is offline detection, enter the sugar and lactic acid concentrations of the offline detection in time to trigger model iteration and correct the predicted data to the offline monitoring value.

[0288] 5. Obtain the real-time model operation results, as shown in Table 2 below.

[0289] Table 2

[0290]

[0291]

[0292] 6. Based on real-time calculation data, the controller can be guided to perform automatic sugar replenishment operations.

[0293] 7. The verification results of this embodiment are as follows Figure 5 , where Figure A is the prediction result and Figure B is the prediction deviation. The entire cell culture process is predictable, and the predictions are all within 1g / L. Except for the 8th day, they are all within the error range of 0.5g / L.

[0294] Example 3

[0295] This example further uses a 250L disposable reactor to experimentally verify the model described in Example 1. The verification steps refer to Example 2.

[0296] The verification results are shown in Table 6, wherein Figure A is the prediction result and Figure B is the prediction deviation. It can be seen from Table 6 that the model constructed by the present invention can accurately predict the glucose concentration during the culture process.

[0297] In summary, the present invention collects conventional cell culture online control parameters (such as dissolved oxygen level, ventilation rate, and stirring speed, etc.), reactor geometry and representative historical batch data, etc., calculates the cumulative oxygen mass transfer of the reactor online detection data, and corrects the model through model operation and historical data, so as to accurately predict the cumulative sugar consumption, and further calculate the residual glucose concentration in the current culture solution. This prediction method only uses conventional online culture parameters without the need to purchase additional hardware equipment, has a more sensitive prediction accuracy, and the entire cell culture process is predictable. It is suitable for small-scale and large-scale cell culture and has broad application prospects.

[0298] The applicant declares that the above is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention shall fall within the protection scope and disclosure scope of the present invention.

Claims

1. A glucose concentration control method based on cumulative oxygen transfer rate, characterized in that: The glucose concentration control method comprises: S1. Collect a batch of historical process data of the same cell line and calculate the daily reference Gluc / OTR coefficient; the daily reference Gluc / OTR coefficient refers to the ratio of the difference between the cumulative Gluc and the cumulative Lac in the historical reference batch from day i to day i+1 to the corresponding cumulative OTR. ref_day(i) ; S2. Collect online process data of cell culture, calculate cumulative OTR, and multiply the cumulative OTR by the correction factor f day(i) Obtain the corrected cumulative OTR; S3. The daily reference Gluc / OTR coefficient is multiplied by the corrected cumulative OTR to obtain the difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment; the cumulative [Gluc] of the predicted batch at the current moment is the cumulative sugar consumption of the predicted batch from the i-th day to the i+t-th day; the cumulative [Lac] of the predicted batch at the current moment is the cumulative lactic acid consumption of the predicted batch from the i-th day to the i+t-th day; The difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment is calculated according to Formula 12 and Formula 11, and the current moment is based on day i; Formula 12: (Accumulated [Gluc] target_day(i+t) -Accumulation [Gluc] target_day(i) )-(cumulative [Lac] target_day(i+t) -Accumulation [Lac] targct_day(i) )=(Cumulative OTR targct_day(i+t) -Cumulative OTR target_day(i) )'×Ratio ref_day(i) Among them, the accumulation of [Gluc] targetday(i+t) Indicates the instantaneous glucose concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Gluc] targetday(i) It indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch integrated over time and volume; Accumulation [Lac] targetday(i+t) Indicates the instantaneous lactate concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Lac] target_day(i) It indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch is integrated over time and volume; Accumulation [Gluc] target_day(i+t) -Accumulation [Gluc] target_day(i) It indicates that the instantaneous glucose concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative sugar consumption from the i-th day to the i+t-th day; Accumulation [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) It indicates that the instantaneous lactic acid concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative lactic acid consumption from the i-th day to the i+t-th day; (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR; The current cumulative amount of the predicted batch [Lac] is calculated according to formula 11; Formula 11: Lac ref_day(i+1) -Lac ref_day(i) It represents the change of lactic acid concentration from day i to day i+1 of the historical reference batch; Time ref_day(i+1) -Time ref_day(i) It represents the time change of lactic acid concentration from day i to day i+1 of the historical reference batch, in days; t represents the time span from the sampling and testing time of the batch to be predicted on the i-th day to the current time, in days; S4. Calculate the current sugar concentration of the predicted batch using the current accumulated [Gluc] of the predicted batch. If the current sugar concentration of the predicted batch is lower than the target sugar concentration, add glucose until the concentration reaches the set value; The predicted current sugar concentration of the batch is calculated according to formula 13; Formula 13: In the formula, Gluc i+t Indicates the current sugar concentration; V represents the system culture volume; [Gluc] i It represents the sugar concentration of the batch to be predicted when sampling and testing on the i-th day; It represents the total amount of sugar supplement from day i to day i+t of the batch to be predicted; It represents the total sugar loss of the batch to be predicted from the i-th day to the i+t-th day manually sampled.

2. The glucose concentration control method based on cumulative oxygen transfer rate according to claim 1, characterized in that: In S1, the daily reference Gluc / OTR coefficient is calculated according to Formula 9; Formula 9: Among them, the accumulation of [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i+1) represents the instantaneous glucose concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the historical reference batch, integrated over time and volume; cumulative [Lac] ref_day(i+1) represents the instantaneous lactate concentration integrated over time and volume from day 0 to day i+1 of the historical reference batch; Accumulation [Gluc] ref_day(i+1) -Accumulation [Gluc] ref_day(i) represents the instantaneous glucose concentration from day i to day i+1 of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i+1) -Accumulation [Lac] ref_day(i) It represents the instantaneous lactic acid concentration from day i to day i+1 of the historical reference batch integrated over time and volume; Cumulative OTR ref_day(i+1) It represents the integration of instantaneous OTR over time and volume from day 0 to day i+1 of the historical reference batch; Cumulative OTR ref_day(i) It represents the integration of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch.

3. The glucose concentration control method based on cumulative oxygen transfer rate according to claim 1 or 2, characterized in that: In S2, the cumulative OTR is the integral value of the instantaneous OUR in the time period from t0 to t, and the cumulative OTR is calculated according to Formula 6; Wherein, V represents the culture volume of the system; Preferably, the instantaneous OUR is calculated according to Formula 5; OTR=OUR=K L a×(DO * -DO) Formula 5 Where OUR represents the instantaneous oxygen transfer rate, K L a represents the gas mass transfer coefficient, DO* represents the saturated dissolved oxygen level, and DO represents the dissolved oxygen level; Preferably, the DO is calculated according to Formula 4; Among them, v O2 is the oxygen flow rate, v air is the air velocity; Preferably, the K L a is calculated according to formula 3; Among them, k, α, β are constants of empirical formula, W s represents the apparent linear velocity, P / V represents the output power per unit volume; Preferably, the W s Calculate according to formula 2; Among them, v O2 is the oxygen flow rate, v air is the air flow rate, Di is the reactor diameter; Preferably, the P / V is calculated according to Formula 1; Among them, N p is the reactor stirring power number, Di is the reactor diameter, and N is the stirring speed.

4. The glucose concentration control method based on cumulative oxygen transfer rate according to any one of claims 1 to 3, characterized in that: In S2, the correction coefficient f day(i) Indicates the deviation correction coefficient between the real-time Gluc / OTR coefficient of the batch to be predicted and the Gluc / OTR coefficient of the reference batch; Preferably, the correction factor f day(i) Calculated according to formula 7; Formula 7: Wherein, i is greater than j, and j is selected from a positive integer less than 5; Accumulation [Gluc] target_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch, integrating time and volume, and accumulating [Gluc] target_day(i-j) It indicates the instantaneous glucose concentration from day 0 to day ij of the predicted batch integrated over time and volume; Accumulation [Lac] target_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch, integrated over time and volume, cumulative [Lac] target_day(i-j) It indicates the instantaneous lactate concentration from day 0 to day ij of the predicted batch integrated over time and volume; Cumulative OTR target_day(i) It represents the integral of instantaneous OTR over time and volume from day 0 to day i of the batch to be predicted; Cumulative OTR target_day(i-j) It represents the integral of instantaneous OTR over time and volume from day 0 to day ij of the batch to be predicted; Accumulation [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day 0 to day ij of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i) Indicates the instantaneous lactic acid concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Lac] ref_day(i-j) represents the instantaneous lactate concentration from day 0 to day ij of the historical reference batch integrated over time and volume; Accumulation [Gluc] ref_day(i) -Accumulation [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day ij to day i of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i) -Accumulation [Lac] ref_day(i-j) It represents the instantaneous lactic acid concentration from day ij to day i of the historical reference batch integrated over time and volume; Cumulative OTR ref_day(i) represents the integral of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch; Cumulative OTR ref_day(i-j) represents the integral of instantaneous OTR over time and volume from day 0 to day ij of the historical reference batch; Preferably, the corrected cumulative OTR is calculated according to Formula 8; Formula 8: (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )=(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )×f day(i) Among them, (cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR.

5. A model for online monitoring of glucose concentration in a cell culture process, characterized in that: The model includes: The historical process data collection unit is used to collect historical process data of a batch of the same cell line and calculate the daily reference Gluc / OTR coefficient; the daily reference Gluc / OTR coefficient refers to the ratio of the difference between the cumulative Gluc and the cumulative Lac in the historical reference batch from the i-th day to the i+1-th day to the corresponding cumulative OTR. ref_day(i) ; Online process data collection unit, used to collect online process data of cell culture, calculate cumulative OTR, and multiply the cumulative OTR by the correction factor f day(i) Obtain the corrected cumulative OTR; The cumulative [Gluc] calculation unit of the predicted batch at the current moment, wherein the daily reference Gluc / OTR coefficient is multiplied by the corrected cumulative OTR to obtain the difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment; the cumulative [Gluc] of the predicted batch at the current moment is the cumulative sugar consumption of the predicted batch from the i-th day to the i+t-th day; the cumulative [Lac] of the predicted batch at the current moment is the cumulative lactic acid consumption of the predicted batch from the i-th day to the i+t-th day; The difference between the cumulative [Gluc] and the cumulative [Lac] of the predicted batch at the current moment is calculated according to Formula 12 and Formula 11, and the current moment is based on day i; Formula 12: (Accumulated [Gluc] target_day(i+t) -Accumulation [Gluc] target_day(i) )-(cumulative [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) )=(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'×Ratio ref_day(i) Among them, the accumulation of [Gluc] targetday(i+t) Indicates the instantaneous glucose concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Gluc] target_day(i) It indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch integrated over time and volume; Accumulation [Lac] target_day(i+t) Indicates the instantaneous lactate concentration from day 0 to day i+t of the predicted batch integrated over time and volume; cumulative [Lac] target_day(i) It indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch is integrated over time and volume; Accumulation [Gluc] target_day(i+t) -Accumulation [Gluc] target_day(i) It indicates that the instantaneous glucose concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative sugar consumption from the i-th day to the i+t-th day; Accumulation [Lac] target_day(i+t) -Accumulation [Lac] target_day(i) It indicates that the instantaneous lactic acid concentration from the i-th day to the i+t-th day of the predicted batch is integrated with time and volume, and the integral value obtained is the cumulative lactic acid consumption from the i-th day to the i+t-th day; (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the corrected cumulative OTR; The current cumulative amount of the predicted batch [Lac] is calculated according to formula 11; Formula 11: Lac ref_day(i+1) -Lac ref_day(i) It represents the change of lactic acid concentration from day i to day i+1 of the historical reference batch; Time ref_day(i+1) -Time ref_day(i) It represents the time change of lactic acid concentration from day i to day i+1 of the historical reference batch, in days; t represents the time span from the sampling and testing time of the batch to be predicted on the i-th day to the current time, in days; a glucose concentration prediction calculation unit, which calculates the current sugar concentration of the predicted batch by using the current accumulated [Gluc] of the predicted batch, and if the current sugar concentration of the predicted batch is lower than the target sugar concentration, adds glucose until the concentration reaches the set value; The predicted current sugar concentration of the batch is calculated according to formula 13; Formula 13: In the formula, Gluc i+t Indicates the current sugar concentration; V represents the system culture volume; [Gluc] i It represents the sugar concentration of the batch to be predicted when sampling and testing on the i-th day; It represents the total amount of sugar supplement from day i to day i+t of the batch to be predicted; It represents the total sugar loss of the batch to be predicted from the i-th day to the i+t-th day manually sampled.

6. The model for online monitoring of glucose concentration in a cell culture process according to claim 5, characterized in that: In the historical process data collection unit, the daily reference Gluc / OTR coefficient is calculated according to Formula 9; Formula 9: Among them, the accumulation of [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i+1) represents the instantaneous glucose concentration from day 0 to day i+1 of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the historical reference batch, integrated over time and volume; cumulative [Lac] ref_day(i+1) represents the instantaneous lactate concentration integrated over time and volume from day 0 to day i+1 of the historical reference batch; Accumulation [Gluc] ref_day(i+1) -Accumulation [Gluc] ref_day(i) represents the instantaneous glucose concentration from day i to day i+1 of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i+1) -Accumulation [Lac] ref_day(i) It represents the instantaneous lactic acid concentration from day i to day i+1 of the historical reference batch integrated over time and volume; Cumulative OTR ref_day(i+1) It represents the integration of instantaneous OTR over time and volume from day 0 to day i+1 of the historical reference batch; Cumulative OTR ref_day(i) It represents the integration of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch.

7. The model for online monitoring of glucose concentration in cell culture process according to claim 5 or 6, characterized in that: In the online process data collection unit, the cumulative OTR is the integral value of the instantaneous OUR in the time period from t0 to t, and the cumulative OTR is calculated according to Formula 6; Wherein, V represents the culture volume of the system; Preferably, the instantaneous OUR is calculated according to Formula 5; OTR=OUR=K L a×(DO * -DO) Formula 5 Where OUR represents the instantaneous oxygen transfer rate, K L a represents the gas mass transfer coefficient, DO* represents the saturated dissolved oxygen level, and DO represents the dissolved oxygen level; Preferably, the DO is calculated according to Formula 4; Among them, v O2 is the oxygen flow rate, v air is the air velocity; Preferably, the K L a is calculated according to formula 3; Among them, k, α, β are constants of empirical formula, W s represents the apparent linear velocity, P / V represents the output power per unit volume; Preferably, the W s Calculate according to formula 2; Among them, v O2 is the oxygen flow rate, v air is the air flow rate, Di is the reactor diameter; Preferably, the P / V is calculated according to Formula 1; Among them, N p is the reactor stirring power number, Di is the reactor diameter, and N is the stirring speed.

8. The model for online monitoring of glucose concentration in a cell culture process according to any one of claims 5 to 7, characterized in that: In the online process data collection unit, the correction factor f day(i) Indicates the deviation correction coefficient between the real-time Gluc / OTR coefficient of the batch to be predicted and the Gluc / OTR coefficient of the reference batch; Preferably, the correction factor f day(i) Calculated according to formula 7; Formula 7: Wherein, i is greater than j, and j is selected from a positive integer less than 5; Accumulation [Gluc] target_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the predicted batch, integrating time and volume, and accumulating [Gluc] target_day(i-j) It indicates the instantaneous glucose concentration from day 0 to day ij of the predicted batch integrated over time and volume; Accumulation [Lac] target_day(i) Indicates the instantaneous lactate concentration from day 0 to day i of the predicted batch, integrated over time and volume, cumulative [Lac] target_day(i-j) It indicates the instantaneous lactate concentration from day 0 to day ij of the predicted batch integrated over time and volume; Cumulative OTR target_day(i) It represents the integral of instantaneous OTR over time and volume from day 0 to day i of the batch to be predicted; Cumulative OTR target_day(i-j) It represents the integral of instantaneous OTR over time and volume from day 0 to day ij of the batch to be predicted; Accumulation [Gluc] ref_day(i) Indicates the instantaneous glucose concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day 0 to day ij of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i) Indicates the instantaneous lactic acid concentration from day 0 to day i of the historical reference batch, integrated over time and volume, cumulative [Lac] ref_day(i-j) represents the instantaneous lactate concentration from day 0 to day ij of the historical reference batch integrated over time and volume; Accumulation [Gluc] ref_day(i) -Accumulation [Gluc] ref_day(i-j) represents the instantaneous glucose concentration from day ij to day i of the historical reference batch integrated over time and volume; Accumulation [Lac] ref_day(i) -Accumulation [Lac] ref_day(i-j) It represents the instantaneous lactic acid concentration from day ij to day i of the historical reference batch integrated over time and volume; Cumulative OTR ref_day(i) represents the integral of instantaneous OTR over time and volume from day 0 to day i of the historical reference batch; Cumulative OTR ref_day(i-j) represents the integral of instantaneous OTR over time and volume from day 0 to day ij of the historical reference batch; Preferably, the corrected cumulative OTR is calculated according to Formula 8; Formula 8: (Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )'=(Cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )×f day(i) Among them, (cumulative OTR target_day(i+t) -Cumulative OTR target_day(i) )' represents the real-time corrected cumulative OTR.

9. A computer-readable storage medium having a computer program / instruction stored thereon, characterized in that: When the computer program / instructions are executed by a processor, the steps in the glucose concentration control method based on cumulative oxygen transfer rate described in any one of claims 1 to 4 are implemented.

10. A computer device comprising a memory and a processor, wherein the memory stores a computer program / instruction, wherein: When the computer program / instructions are executed by a processor, the steps in the glucose concentration control method based on cumulative oxygen transfer rate described in any one of claims 1 to 4 are implemented.

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