Method for developing and characterizing blending process of biological preparation stock solution and application of method
Through constant average power input and computational fluid mechanics simulation, the mixing process of biological preparation stock solution is optimized, and the problem of inconsistent quality and efficacy in the mixing process of biological preparation stock solution is solved, achieving efficient and reliable mixing effect and product consistency.
Patent Information
- Application Number
- CN202510452104.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-11
- Publication Date
- 2025-07-11
AI Technical Summary
The lack of systematic development and characterization strategies during the mixing of biological agent stock solutions, resulting in inconsistent drug quality and efficacy. The existing technology relies on rules of thumb and lacks scientific design, making it difficult to ensure uniformity and protect the structural integrity of sensitive biological materials.
A constant average power input strategy is adopted, combined with computational fluid mechanics simulation, and the biological preparation stock liquid mixing process is developed. By simulating liquid volume, temperature, viscosity and equipment changes, we ensure that the conductivity of each position in the mixing system is consistent. We use conductive solution as an indicator to optimize the sampling position and power input.
The uniformity and structural integrity of the biological preparation stock solution within the conventional mixing process time is achieved, the consistency of production efficiency and product quality is improved, the damage to biological materials by shear forces is reduced, and the cost and time is reduced.
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Figure CN120294277A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biological medicine technology, and in particular to a method for the development and characterization of a biological agent stock solution mixing process and its application. Background Art
[0002] Biological agents have become an important part of the current pharmaceutical industry. Among them, antibody drugs have shown significant efficacy in the treatment of various diseases including cancer, autoimmune diseases, and inflammatory diseases.
[0003] Stock solution mixing is a key step in the production process of biological agents. The stock solution is usually in a frozen state to improve its long-term storage stability and extend its shelf life. Therefore, the frozen stock solution needs to be thawed before the filling / finishing process. However, the components of the thawed stock solution are unevenly distributed in the storage container, so it must be transferred to a mixing system for mixing operations to ensure the uniform distribution of the stock solution. From laboratory scale to commercial production, the mixing process is the key to ensuring the quality and efficacy consistency of the final biological agent product.
[0004] In the production process of biological agents, different batches of stock solutions, different mixing systems, and stock solutions with different viscosities and temperatures are often involved. Different batch productions require the mixing system to give appropriate power input to ensure thorough mixing of the stock solution. At the same time, an appropriate mixing system needs to be selected because the geometric properties of the mixing system also affect the mixing efficiency. Meanwhile, high-viscosity or low-temperature stock solutions will bring more viscous resistance, hindering the movement of the stirring paddle in the mixing system. All of the above factors will affect the hydrodynamic behavior and homogeneity of the stock solution in the mixing system, thus affecting the product quality. Higher power input is beneficial to the mixing of the stock solution, but excessive power input will introduce higher shear forces. And biological agents have complex structures, which are crucial for their biological activities. Therefore, an appropriate stock solution mixing process can not only ensure the uniformity of the stock solution, but also maintain the structural integrity of these sensitive molecules, avoiding the loss of activity caused by excessive shear forces or improper handling.
[0005] A good stock solution mixing process can ensure the uniform distribution of all components throughout the batch, which is crucial for producing drugs with consistent quality and efficacy. The current development of the biological agent stock solution mixing process largely depends on empirical rules and lacks a clear development strategy. And in the characterization of the stock solution mixing process, multiple dense sampling points are usually set to characterize the mixing situation of the solution, but this approach often lacks a scientific design basis.
[0006] In summary, the development and characterization of the stock solution mixing process are important aspects of the research and production of biological products. Currently, there is a lack of a consensus and systematic strategy for the development and characterization of the stock solution mixing process in the biopharmaceutical industry, which poses challenges to ensuring the quality and efficacy consistency of drugs. Therefore, it is urgent to develop a strategy for the development and characterization of the stock solution mixing process applicable to biological preparations to ensure the homogeneity of the stock solution in the filling / finishing process, thereby producing drugs with consistent quality and efficacy. This not only helps improve production efficiency but also ensures that patients receive safe and effective treatments. Summary of the Invention
[0007] To solve the above technical problems, the present invention provides a method and its application for the development and characterization of the stock solution mixing process of biological preparations. The development of the stock solution mixing process of biological preparations is carried out by means of a strategy of constant average power input, taking into account different average power inputs, stock solutions with different viscosities, stock solutions at different temperatures, batch changes, equipment changes, etc. that may occur in the manufacture of biological preparations, and combining with the conventional production duration, and finally producing biological preparations with consistent quality and efficacy.
[0008] To achieve this purpose, the present invention adopts the following technical solutions:
[0009] In the first aspect, the present invention provides a method for the development and characterization of the stock solution mixing process of biological preparations, and the method for the development and characterization of the stock solution mixing process of biological preparations includes the following steps:
[0010] (1) Prepare a first simulated solution, a second simulated solution, and a conductive solution, where the viscosities of the first simulated solution and the second simulated solution are different;
[0011] (2) Place the first simulated solution in a mixing system, add the conductive solution, preset a unit volume average power input value, and operate the stirring system at the unit volume average power with the temperature of the simulated solution being 15 - 30 °C to stir and mix the mixing system, detect the conductivity, and record the mixing time until the conductivity values at each position tend to be consistent. If the mixing time is greater than the conventional mixing process duration (i.e., greater than 40 min), then change the preset value of the unit volume average power and repeat step (2);
[0012] (3) Replace the first simulated solution in step (2) with the second simulated solution and repeat step (2) again;
[0013] (4) Control the temperature of the simulated solution in step (2) at 2 - 8 °C and repeat step (2) again;
[0014] (5) Change the volume of the first simulated solution in the mixing system in step (2) and repeat step (2) again;
[0015] (6) Change the type of the mixing system in step (2), and perform step (2) again to obtain the average power input value per unit volume at this time, which is the average power per unit volume during the mixing of the biopreparation stock solution.
[0016] The specific point values of 15 - 30°C above can be 15°C, 16°C, 18°C, 20°C, 22°C, 24°C, 25°C, 26°C, 28°C or 30°C, etc.
[0017] The specific point values of 2 - 8°C above can be 2°C, 3°C, 4°C, 5°C, 6°C, 7°C or 8°C, etc.
[0018] In the biopreparation cell culture process, the development strategy based on the average power input has been widely applied. This strategy can ensure mixing uniformity and appropriate shear force to protect sensitive biological materials. The cell culture process involves the addition of culture medium and the introduction of oxygen, requiring a relatively high average power input (generally 10 - 40 W / m 3 ) to quickly mix the supplemented culture medium and oxygen within a short time and maintain the uniform distribution of components throughout the cell culture cycle (for example, when producing antibodies using Chinese hamster ovary cells, the culture cycle is generally 10 - 14 days).
[0019] The purpose of the stock solution mixing process is to mix the thawed stock solution with uneven components within the conventional mixing operation duration (such as 20 - 40 minutes). Therefore, compared with the cell culture process, the average power input can be appropriately reduced for stock solution mixing, while reducing the damage of shear force to biological materials during the process. Therefore, it is necessary to develop a suitable average power input range for the stock solution mixing process. When developing, different average power inputs, stock solutions with different viscosities, different stock solution temperatures, batch changes, equipment changes, etc. that may exist in actual production need to be considered to ensure that the developed average power input range can achieve the component uniformity of the thawed stock solution within the conventional mixing process duration.
[0020] The present invention conducts process development based on the average power input, guides multi - batch production (assuming comparability between batches), and is applicable to different average power inputs, stock solutions with different viscosities, different stock solution temperatures, batch changes, equipment changes, etc. that may occur in biopreparation production, and combines the conventional production duration to finally produce antibody drugs with consistent quality and efficacy.
[0021] Preferably, the first simulation liquid and the second simulation liquid each independently contain a buffer, a surfactant, and an adjuvant.
[0022] The stock solution cost of biologic agents is relatively high and the materials are relatively scarce. It is usually difficult to develop and characterize the mixing process using a large volume of the stock solution of biologic agents. Therefore, the present invention uses a simulated solution to develop and characterize the mixing process of the stock solution of biologic agents, and the simulated solution can be placed at room temperature or 2 - 8°C according to the actual production situation.
[0023] Preferably, the buffer solution includes any one or a combination of at least two of histidine buffer solution, citrate buffer solution, acetate buffer solution, succinate buffer solution, phosphate buffer solution, or tris(hydroxymethyl)aminomethane buffer solution.
[0024] Preferably, the surfactant includes any one or a combination of at least two of polysorbate 20, polysorbate 80, or poloxamer 188.
[0025] Preferably, the excipient includes any one or a combination of at least two of polyethylene glycol 4000, polyethylene glycol 6000, or polyethylene glycol 10000.
[0026] The present invention adds polyethylene glycol excipient to the simulated solution to simulate the viscosity conditions of the stock solution of biologic agents, and even higher viscosity situations.
[0027] Preferably, the excipient further includes any one or a combination of at least two of sucrose, sorbitol, or trehalose.
[0028] Preferably, the conductive solution contains an electrolyte.
[0029] Preferably, the electrolyte includes sodium chloride and / or arginine hydrochloride.
[0030] Preferably, after adding the conductive solution, the mixing system contains 5 - 100 mM histidine buffer solution, 0 - 600 mg / mL polyethylene glycol 6000, 0.01% - 0.2% w / v polysorbate 80, and 50 - 300 mM sodium chloride, and the pH is 4 - 8.
[0031] The specific point values of the above 5 - 100 mM can be 5 mM, 7 mM, 10 mM, 15 mM, 20 mM, 40 mM, 60 mM, 80 mM, or 100 mM, etc.
[0032] The specific point values of the above 0 - 600 mg / mL can be 0 mg / mL, 1 mg / mL, 5 mg / mL, 10 mg / mL, 50 mg / mL, 100 mg / mL, 200 mg / mL, 300 mg / mL, 400 mg / mL, 500 mg / mL, or 600 mg / mL, etc.
[0033] The specific point values of the above 0.01% - 0.2% can be 0.01%, 0.02%, 0.04%, 0.07%, 0.1%, 0.15%, 0.17% or 0.2%, etc.
[0034] The specific point values of the above 50 - 300 mM can be 50 mM, 60 mM, 80 mM, 100 mM, 150 mM, 200 mM, 250 mM or 300 mM, etc.
[0035] The specific point values of the above 4 - 8 can be 4, 5, 5.5, 6, 6.5, 7 or 8, etc.
[0036] Among them, 20 mM histidine buffer (pH 6.0) is a commonly used buffer system in marketed biological agents, sodium chloride and polysorbate 80 are commonly used excipients and surfactants in biological agents, and polyethylene glycol 6000 is a mixture formed by the polycondensation of ethylene oxide and water, which is used to increase the viscosity of the simulation liquid.
[0037] Preferably, the method for developing and characterizing the mixing process of the biological agent stock solution further includes: identifying the positions in the mixing system that are more difficult to mix and those that are easier to mix through computational fluid dynamics simulation.
[0038] The present invention uses computational fluid dynamics simulation technology to identify multiple key positions in the mixing system that are difficult to mix, and sets them as representative sampling points. By using a high - concentration conductive solution (such as sodium chloride solution) as an indicator, it is added to the simulation liquid from one of the positions that are more difficult to mix. At the same time, the constant average input power in the computational fluid dynamics simulation is maintained to set the mixing parameters matching the volume of the simulation liquid, sampling is carried out at the representative sampling positions, and the conductivity values of the solutions at each location are detected. When the conductivity values of the solutions at each location tend to be consistent, it indicates that the simulation liquid has achieved uniform distribution in the mixing system.
[0039] Preferably, when the mixing system is a cylindrical mixing system, the positions that are more difficult to mix include the side walls of the liquid surface, the center of the liquid surface, and the positions near the lower part of the two side walls, and the positions that are easier to mix include the vicinity of the stirring paddle.
[0040] Preferably, when the mixing system is a cubic - shaped mixing system, the positions that are more difficult to mix include the side walls of the liquid surface, the center of the liquid surface, the bottom corners, and the center of the side walls, and the positions that are easier to mix include the vicinity of the stirring paddle.
[0041] Preferably, the method for adding the conductive solution includes: adding the conductive solution from at least one position that is more difficult to mix.
[0042] Preferably, the method for detecting the conductivity includes: simultaneously detecting the conductivity of at least one position that is more difficult to mix and at least one position that is easier to mix.
[0043] Preferably, the types of the mixing system in the changing step (2) specifically include: changing the specifications of the mixing system and / or changing the geometric shape of the mixing system.
[0044] The types of the mixing system referred to in the present invention can change the geometric shape of the mixing system. For example, changing a cylindrical mixing system to a cubic mixing system; it can also change the specifications of the mixing system. Similarly, for a cylindrical mixing system, there are various specifications such as 50L / 100L / 200L / 500L / 1000L. For example, changing a 200L mixing system to a 500L mixing system.
[0045] In a second aspect, the present invention provides a model for the development and characterization of a biological agent stock solution mixing process, and the model for the development and characterization of a biological agent stock solution mixing process is used to execute the method for the development and characterization of a biological agent stock solution mixing process described in the first aspect.
[0046] Preferably, the model for the development and characterization of a biological agent stock solution mixing process includes:
[0047] Solution preparation module: used for preparing the first simulation solution, the second simulation solution and the conductive solution;
[0048] Solution addition module: used for placing the first simulation solution or the second simulation solution into the mixing system and adding the conductive solution;
[0049] Mixing system adjustment module: used for adjusting the temperature of the simulation solution, adjusting the volume of the first simulation solution in the mixing system, changing the types of the mixing system, and stirring the mixing system with a constant average power per unit volume;
[0050] Conductivity detection module: used for detecting the conductivity of the mixing system until the conductivity values at each position tend to be consistent, recording the mixing time. If the mixing time is longer than the conventional mixing process duration (i.e., longer than 40 min), then feedback to the mixing system module to change the average power per unit volume.
[0051] Preferably, the model for the development and characterization of a biological agent stock solution mixing process further includes a mixing system identification module, and the mixing system identification module is used to identify the positions in the mixing system that are difficult to mix and easy to mix through computational fluid dynamics simulation.
[0052] Preferably, the solution addition module is specifically used for placing the first simulation solution or the second simulation solution into the mixing system and adding the conductive solution into the mixing system from at least one position that is difficult to mix.
[0053] Preferably, the conductivity detection module is specifically configured to simultaneously detect the conductivities of at least one position that is difficult to mix evenly and at least one position that is easy to mix evenly among the identified positions until the conductivity values at each position tend to be consistent, record the mixing time. If the mixing time is greater than the conventional mixing process duration (i.e., greater than 40 minutes), then feedback to the mixing system module to change the average power per unit volume.
[0054] In a third aspect, the present invention provides the method for developing and characterizing the bio - preparation stock solution mixing process as described in the first aspect and / or the application of the bio - preparation stock solution mixing process development and characterization model as described in the second aspect in large - scale bio - preparation production.
[0055] Other specific point values within the above - mentioned numerical ranges can be selected, and will not be elaborated one by one here.
[0056] Compared with the prior art, the present invention has the following beneficial effects:
[0057] The development strategy of the bio - preparation stock solution mixing process proposed by the present invention is based on a constant average power input, ensuring the comparability between batches when the same bio - preparation is produced in different batches or by different mixing devices. Through computational fluid dynamics simulation, it accurately guides the setting of sampling positions. Compared with the traditional intensive sampling method, it significantly reduces time and cost, while improving the representativeness of sampling. This strategy not only optimizes the bio - preparation stock solution mixing process, but also provides a reliable guarantee for ensuring the consistency of product quality during the production process, thereby enhancing the production and R & D efficiency of bio - preparations and product quality control. Brief Description of the Drawings
[0058] Figure 1 Schematic diagram of positions that are difficult to mix evenly and easy to mix evenly in the cylindrical mixing system in Example 3.
[0059] Figure 2 Representative sampling position diagram of the cylindrical mixing system in the stock solution mixing process characterization in Example 3.
[0060] Figure 3 Schematic diagram of positions that are difficult to mix evenly and easy to mix evenly in the cubic - shaped mixing system in Example 4.
[0061] Figure 4 Representative sampling position diagram of the cubic - shaped mixing system in the stock solution mixing process characterization in Example 4.
[0062] Figure 5 For Example 5, the conductivity change trend diagram of each sampling position at an average power input of 3W / m 3
[0063] Figure 6 For Example 5, the conductivity change trend diagram of each sampling position at an average power input of 5W / m 3Conductivity change trend diagram for each sampling position.
[0064] Figure 7 It is the conductivity change trend diagram for each sampling position of the first simulated liquid in Example 6.
[0065] Figure 8 It is the conductivity change trend diagram for each sampling position of the second simulated liquid in Example 6.
[0066] Figure 9 It is the conductivity change trend diagram for each sampling position when the solution temperature is 4°C in Example 7.
[0067] Figure 10 It is the conductivity change trend diagram for each sampling position of the 200L simulated liquid in Example 8.
[0068] Figure 11 It is the conductivity change trend diagram for each sampling position of the 500L simulated liquid in Example 8. Detailed implementation manners
[0069] To further elaborate on the technical means and effects adopted by the present invention, the present invention will be further described below in conjunction with examples and drawings. It can be understood that the specific implementation manners described herein are only used to explain the present invention, rather than limiting the present invention.
[0070] For those not specifying specific techniques or conditions in the examples, they shall be carried out according to the techniques or conditions described in the literature in this field or according to the product specifications. For those reagents or instruments not specifying the manufacturer, they are all conventional products that can be obtained by purchasing through regular channels.
[0071] Example 1
[0072] This example provides a method for the development and characterization of a biological agent stock solution mixing process. The method for the development and characterization of the biological agent stock solution mixing process includes the following steps:
[0073] (1) Prepare the first simulated liquid, the second simulated liquid, and the conductive solution.
[0074] (2) Place the first simulated liquid in a mixing system, add the conductive solution, preset a unit volume average power input value, operate the stirring system at the unit volume average power at 25°C, stir and mix the mixing system, detect the conductivity until the conductivity values at each position tend to be consistent, record the mixing time. If the mixing time is greater than 40 min, change the preset value of the unit volume average power and repeat step (2);
[0075] After adding the conductive solution, the mixing system contains 20 mM histidine buffer, 180 mg / mL polyethylene glycol 6000, 0.02% w / v polysorbate 80, and 140 mM sodium chloride, with a pH of 6 and a viscosity of 10.01 cP at 20 °C.
[0076] (3) Replace the first simulation liquid in step (2) with the second simulation liquid and perform step (2) again.
[0077] After adding the conductive solution, the mixing system contains 20 mM histidine buffer, 0.02% w / v polysorbate 80, and 140 mM sodium chloride, with a pH of 6 and a viscosity of 2.15 cP at 20 °C.
[0078] (4) Control the temperature of the simulation liquid in step (2) to 4 °C and perform step (2) again.
[0079] (5) Change the volume of the first simulation liquid in the mixing system of step (2) and perform step (2) again.
[0080] (6) Change the type of the mixing system in step (2) and perform step (2) again to obtain the average power input value per unit volume at this time, which is the average power per unit volume during the mixing of the original biological preparation.
[0081] Before adding the conductive solution in steps (2) to (6), through computational fluid dynamics simulation, identify the positions in the mixing system that are difficult to mix and easy to mix, add the conductive solution from at least one position that is difficult to mix, and simultaneously detect the conductivity of at least one position that is difficult to mix and at least one position that is easy to mix.
[0082] Example 2
[0083] This example provides a model for the development and characterization of a mixing process for the original biological preparation based on Example 1, including the following modules:
[0084] Solution preparation module: used to prepare the first simulation liquid, the second simulation liquid, and the conductive solution;
[0085] Solution addition module: used to place the first simulation liquid or the second simulation liquid in the mixing system and add the conductive solution to the mixing system from at least one position that is difficult to mix;
[0086] Mixing system identification module: used to identify the positions in the mixing system that are difficult to mix and easy to mix through computational fluid dynamics simulation;
[0087] Mixing system adjustment module: used to adjust the temperature of the simulation liquid, adjust the volume of the first simulation liquid in the mixing system, change the type of the mixing system, and stir the mixing system with a constant average power per unit volume;
[0088] Conductivity detection module: used to detect the conductivity of at least one position that is difficult to mix evenly and at least one position that is easy to mix evenly simultaneously until the conductivity values of each position tend to be consistent, record the mixing time, and if the mixing time is greater than 40 minutes, feedback to the mixing system module to change the average power per unit volume.
[0089] Example 3
[0090] This example aims to reveal the positions that are difficult to mix evenly in a cylindrical mixing system through computational fluid dynamics simulation and give suggestions on the sampling positions in the mixing process characterization of the biopharmaceutical stock solution. Figure 1 For the computational fluid dynamics simulation results, Figure 2 For the sampling position suggestions. Positions 1 - 4 (Position 1 - 4) are the 4 positions that are difficult to mix evenly in the same plane of the cylindrical mixing system identified based on computational fluid dynamics simulation, namely the liquid surface sidewall, the liquid surface center, and the positions near the lower part of the two sidewalls. Position 5 (Position 5) is the position that is easy to mix evenly in the cylindrical mixing system, located near the stirring paddle. In the mixing process characterization of the stock solution, samples are taken regularly at positions 1 - 5. When the conductivity of the simulated liquid at all positions tends to be consistent, it is considered that the simulated liquid has reached the mixed state. It is recommended to take samples of positions 1 - 4 at different planes, such as the plane where positions 1 - 2 are located intersects perpendicularly with the plane where positions 3 - 4 are located.
[0091] Example 4
[0092] This example aims to reveal the positions that are difficult to mix evenly in a cubic mixing system through computational fluid dynamics simulation and give suggestions on the sampling positions in the mixing process characterization of the biopharmaceutical stock solution. Figure 3 For the computational fluid dynamics simulation results, Figure 4 For the sampling position suggestions. The liquid surface center (Top - 1), the liquid surface sidewall (Top - 2), the bottom corner (Bottom - 5), and the sidewall center (Middle - 3) are the 4 positions that are difficult to mix evenly in the cubic mixing system identified based on computational fluid dynamics simulation. The position near the stirring paddle (Bottom - 4) is the position that is easy to mix evenly in the cubic mixing system. In the mixing process characterization of the stock solution, samples are taken regularly at the above 5 positions. When the conductivity of the simulated liquid at all positions tends to be consistent, it is considered that the simulated liquid has reached the mixed state.
[0093] Example 5
[0094] Development of the mixing process of simulated liquids (solution temperature is 25°C) with the same volume and viscosity under different average input powers in a 50L Merck mixing system.
[0095] In this example, 50 L of simulated liquid was added to a 50 L Merck mixing system. Through computational fluid dynamics simulation, the addition position of the conductive solution and the conductivity detection position were determined. After adding the conductive solution, the mixing system contained 20 mM histidine buffer, 180 mg / mL polyethylene glycol 6000, 0.02% w / v polysorbate 80, and 140 mM sodium chloride, with a pH of 6 and a viscosity of 10.01 cP at 20 °C, being a Newtonian fluid. At 25 °C, for the mixing system to maintain average power inputs of 3 W / m 3 and 5 W / m 3 , stirring speeds of 250 rpm and 300 rpm were respectively required. Process development was carried out according to the simulated mixing speed, and the mixing situation of the solution was detected using conductivity as an indicator.
[0096] Figure 5 For controlling the average power input to 3 W / m 3 in a 50 L Merck mixing system for a simulated liquid with a viscosity of 10.01 cP (20 °C) (solution temperature 25 °C), it is the conductivity change trend graph at each sampling position, Figure 6 and for controlling the average power input to 5 W / m 3 in a 50 L Merck mixing system for a simulated liquid with a viscosity of 10.01 cP (20 °C) (solution temperature 25 °C), it is the conductivity change trend graph at each sampling position. The results showed that it took 25 minutes and 15 minutes respectively to achieve mixing, and the conductivity at each sampling position tended to be consistent. Based on this example, it is recommended that during the mixing process development of biopharmaceutical stock solutions, in combination with the conventional mixing process duration of production, the range of appropriate average power inputs be studied.
[0097] Example 6
[0098] In a 200 L Merck mixing system, for simulated liquids with different viscosities but the same volume (solution temperature 25 °C), the mixing process was developed while maintaining the same average input power.
[0099] This example aimed to develop a mixing process for stock solutions with different viscosities by controlling the viscosity of the simulated liquid. Two simulated liquids with different viscosities, the first simulated liquid and the second simulated liquid provided in Example 1, were placed in a 200 L Merck mixing system. The volume of the simulated liquid was 200 L, and the average power input was controlled at 10 W / m 3 , that is, a stirring speed of 280 rpm. The operation during the experiment was the same as that in Example 5.
[0100] Figure 7 For controlling the average power input to 10 W / m 3 in a 200 L Merck mixing system for a simulated liquid with a viscosity of 10.01 cP (20 °C) (solution temperature 25 °C), it is the conductivity change trend graph at each sampling position,Figure 8 For controlling the average power input to 10 W / m for a simulated liquid with a viscosity of 2.15 cP (at 20 °C) in a 200 L Merck mixing system (solution temperature is 25 °C). 3 The conductivity change trend graph at each sampling position when the power is input at this level. Figure 7 Among the 9 sampling positions in [], it includes the 5 sampling positions mentioned in Example 3, and additionally samples are taken at 2 positions at the liquid surface sidewall of the mixing system, the system centroid position, and 1 position at the bottom sidewall. Figures 8 - 11 The conductivity detection positions are the same as Figure 7 those described previously.
[0101] The results show that at this average power input, the 200 L stirring system can achieve mixing of simulated liquids with viscosities of 2.15 cP (at 20 °C) and 10.01 cP (at 20 °C) within 5 minutes, indicating that at this constant average power input, during the conventional mixing process duration, stock solutions with different viscosities can all achieve mixing. Therefore, in this example, 10 W / m 3 is set as the target value of the average power input.
[0102] When mixing stock solutions in biopharmaceuticals, there are stock solutions with different viscosities (routinely 1 - 50 cP, at 20 °C). It is time - consuming and laborious to investigate the appropriate average power input for each viscosity of stock solution. Therefore, it is recommended that during the development of the biopharmaceutical stock solution mixing process, the appropriate average power input be investigated so that stock solutions with conventional viscosities can all achieve mixing within the conventional mixing process duration, and the finally investigated appropriate average power input value be set as the production operation target value.
[0103] Example 7
[0104] For stock solutions with the same volume and viscosity at different temperatures in a 200 L Merck mixing system, the mixing process is developed while maintaining the same average input power.
[0105] The purpose of this example is to develop the process for mixing low - temperature stock solutions that may occur in production by controlling the temperature of the simulated liquid. In a 200 L Merck mixing system, for simulated liquids with the same volume and viscosity, the temperature is controlled at 25 °C or 4 °C, and the mixing time is investigated.
[0106] Maintaining the simulated liquid with the same volume and viscosity as in Example 6 (10.01 cP, at 20 °C) and the experimental operation process, the average input power is controlled at 10 W / m 3 . Figure 9 For controlling the average power input to 10 W / m for a simulated liquid with a viscosity of 10.01 cP (at 20 °C) in a 200 L Merck mixing system (solution temperature is 4 °C). 3 The conductivity change trend graph at each sampling position when the power is input at this level.
[0107] By comparing Figure 7 and Figure 9 it can be seen that when the solution is at 25°C or 4°C, it only takes 5 minutes to mix evenly, indicating that under the condition of maintaining the target average power input value set in Example 6, even if the stock solution temperatures vary among different production batches, it is possible to achieve even mixing within the conventional mixing process duration, ensuring comparability among batches. Therefore, when developing the mixing process for biopharmaceutical stock solutions, the influence of solution temperature on mixing uniformity should be investigated, and an appropriate target average power input value should be selected to ensure that stock solutions of the same type at different temperatures can all be evenly mixed within the conventional mixing process duration, guaranteeing comparability among batches.
[0108] Example 8
[0109] For simulated liquids with different volumes but the same viscosity (solution temperature is 25°C), the mixing process was developed in a 500L Merck mixing system while maintaining the same average input power.
[0110] This example aims to develop the mixing process for different batches of stock solutions in production by controlling the volume of the simulated liquid. In a 500L Merck mixing system, the average input power was controlled at 10W / m 3 , the composition of the simulated liquid was the same as that in Example 1, the viscosity was 10.01 cP (20°C), the experimental operations were the same, and the volumes of the simulated liquid were 200L and 500L respectively.
[0111] Figure 10 Figure showing the conductivity change trend at each sampling position when controlling the average power input to 10W / m 3 for a 200L simulated liquid with a viscosity of 10.01 cP (20°C) (solution temperature is 25°C) in a 500L Merck mixing system, Figure 11 Figure showing the conductivity change trend at each sampling position when controlling the average power input to 10W / m 3 for a 500L simulated liquid with a viscosity of 10.01 cP (20°C) (solution temperature is 25°C) in a 500L Merck mixing system. The results show that for simulated liquids with different volumes, they still achieved even mixing in 5 minutes under this parameter, indicating that under the condition of maintaining the target average power input value set in Example 6, with the same mixing equipment, even when the batch size changes, it is possible to achieve even mixing within the conventional mixing process duration, ensuring comparability among batches. It is recommended that when developing the mixing process for biopharmaceutical stock solutions, the influence of batch size change on mixing uniformity under the same equipment be investigated, and an appropriate target average input power be selected to ensure comparability among batches.
[0112] Example 9
[0113] In different mixing systems, simulation liquids (solution temperature is 25°C) with the same volume and the same viscosity are used to develop the mixing process while maintaining the same average input power.
[0114] This embodiment aims to develop the process for possible changes in mixing equipment during production by investigating the mixing performance of different mixing equipment.
[0115] Keep the simulation liquid (10.01 cP, 20°C) with the same components as in Example 1 and the experimental operation process, and control the average input power at 10 W / m 3 , with the volume of the simulation liquid being 200 L. Investigate the mixing time in a 200 L mixing system and a 500 L mixing system respectively. The results show that the 200 L simulation liquid can be mixed within 5 minutes in the 200 L mixing system in Example 6 ( Figure 7 ) and the 500 L mixing system in Example 8 ( Figure 10 ). This indicates that under the target average power input value set in Example 6, for the same volume of the original liquid, even if the mixing equipment is changed, it can still be mixed within the conventional mixing process duration, ensuring comparability between batches. It is recommended that when developing the mixing process of the biological agent original liquid, investigate the influence of equipment performance on mixing uniformity and select an appropriate target average input power to ensure comparability between batches.
[0116] In summary, the present invention develops the mixing process of the biological agent original liquid through the strategy of constant average power input, taking into account various situations that may occur in biological agent manufacturing, such as different average power inputs, original liquids with different viscosities, different original liquid temperatures, batch changes, and equipment changes, and combining with the conventional production duration, finally producing biological agents with consistent quality and efficacy. At the same time, the present invention combines computational fluid dynamics simulation to give suggestions on the representative sampling positions for characterizing the mixing process of the biological agent original liquid.
[0117] The applicant declares that the above is only the specific implementation manner 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 within the technical scope disclosed by the present invention fall within the protection scope and the disclosure scope of the present invention.
Claims
1. A method for the development and characterization of a homogenization process for a biological agent stock solution, characterized in that, The method for developing and characterizing the mixing process of the biopharmaceutical stock solution includes the following steps: (1) Prepare a first simulation solution, a second simulation solution, and a conductive solution, where the viscosities of the first simulation solution and the second simulation solution are different; (2) Place the first simulation solution in a mixing system, add the conductive solution, preset a unit volume average power input value, and operate the stirring system at the unit volume average power with the simulation solution temperature at 15 - 30 °C to stir and mix the mixing system, detect the conductivity, and record the mixing time until the conductivity values at each position tend to be consistent. If the mixing time is longer than the conventional mixing process duration, change the preset value of the unit volume average power and repeat step (2); (3) Replace the first simulation solution in step (2) with the second simulation solution and repeat step (2); (4) Control the temperature of the simulation solution in step (2) at 2 - 8 °C and repeat step (2); (5) Change the volume of the first simulation solution in the mixing system in step (2) and repeat step (2); (6) Change the type of the mixing system in step (2) and repeat step (2) to obtain the unit volume average power input value at this time, which is the unit volume average power during the mixing of the biopharmaceutical stock solution.
2. The method for the development and characterization of the mixing process of the biological agent stock solution according to claim 1, characterized in that, The first simulation solution and the second simulation solution each independently contain a buffer, a surfactant, and an excipient; Preferably, the buffer includes any one or a combination of at least two of histidine buffer, citrate buffer, acetate buffer, succinate buffer, phosphate buffer, or tris(hydroxymethyl)aminomethane buffer; Preferably, the surfactant includes any one or a combination of at least two of polysorbate 20, polysorbate 80, or poloxamer 188; Preferably, the excipient includes any one or a combination of at least two of polyethylene glycol 4000, polyethylene glycol 6000, or polyethylene glycol 10000; Preferably, the excipient further includes any one or a combination of at least two of sucrose, sorbitol, or trehalose; Preferably, the conductive solution contains an electrolyte; Preferably, the electrolyte includes sodium chloride and / or arginine hydrochloride; Preferably, after adding the conductive solution, the mixing system contains 5 - 100 mM histidine buffer, 0 - 600 mg / mL polyethylene glycol 6000, 0.01% - 0.2% w / v polysorbate 80, and 50 - 300 mM sodium chloride, with a pH of 4 - 8.
3. The method for the development and characterization of the mixing process of the biological agent stock solution according to claim 1 or 2, characterized in that, The method for developing and characterizing the mixing process of the biopharmaceutical stock solution further includes: identifying the positions in the mixing system that are difficult to mix and easy to mix through computational fluid dynamics simulation; Preferably, when the mixing system is a cylindrical mixing system, the positions difficult to mix include the liquid surface sidewall, the liquid surface center, and the positions near the lower part of the two sidewalls, and the positions easy to mix include near the stirring paddle; Preferably, when the mixing system is a cubic mixing system, the positions difficult to mix include the liquid surface sidewall, the liquid surface center, the bottom corners, and the center of the sidewalls, and the positions easy to mix include near the stirring paddle.
4. The method for the development and characterization of the mixing process of the biopreparation stock solution according to any one of claims 1 to 3, characterized in that, The adding method of the conductive solution includes: adding the conductive solution from at least one position that is difficult to mix; Preferably, the method for detecting the conductivity includes: simultaneously detecting the conductivity at at least one position that is difficult to mix evenly and at least one position that is easy to mix evenly.
5. The method for the development and characterization of the mixing process of the biopreparation stock solution according to any one of claims 1 to 4, characterized in that, The specific manner of changing the type of the mixing system in step (2) includes: changing the specifications of the mixing system and / or changing the geometric shape of the mixing system.
6. A biological agent stock solution mixing process development and characterization model, characterized in that, The biologic drug substance mixing process development and characterization model is used to execute the method for the development and characterization of the biologic drug substance mixing process according to any one of claims 1 to 5.
7. The biopreparation stock solution mixing process development and characterization model according to claim 6, characterized in that The biologic drug substance mixing process development and characterization model includes: A solution preparation module: used for preparing a first simulation solution, a second simulation solution, and a conductive solution; A solution addition module: used for placing the first simulation solution or the second simulation solution into the mixing system and adding the conductive solution; A mixing system adjustment module: used for adjusting the temperature of the simulation solution, adjusting the volume of the first simulation solution in the mixing system, changing the type of the mixing system, and stirring the mixing system with a constant average power per unit volume; A conductivity detection module: used for detecting the conductivity of the mixing system until the conductivity values at all positions tend to be consistent, recording the mixing time, and if the mixing time is greater than the conventional mixing process duration, feeding back to the mixing system module to change the average power per unit volume.
8. The biologic bulk mixing process development and characterization model according to claim 6 or 7, characterized in that The biologic drug substance mixing process development and characterization model further includes a mixing system identification module, and the mixing system identification module is used to identify the positions that are difficult to mix evenly and the positions that are easy to mix evenly in the mixing system through computational fluid dynamics simulation.
9. The biologic bulk mixing process development and characterization model according to any one of claims 6 to 8, characterized in that, The solution addition module is specifically used for placing the first simulation solution or the second simulation solution into the mixing system and adding the conductive solution into the mixing system from at least one position that is difficult to mix evenly; Preferably, the conductivity detection module is specifically used for simultaneously detecting the conductivity at at least one identified position that is difficult to mix evenly and at least one position that is easy to mix evenly until the conductivity values at all positions tend to be consistent, recording the mixing time, and if the mixing time is greater than the conventional mixing process duration, feeding back to the mixing system module to change the average power per unit volume.
10. The application of the method for the development and characterization of the biologic drug substance mixing process according to any one of claims 1 to 5 and / or the biologic drug substance mixing process development and characterization model according to any one of claims 6 to 9 in large-scale preparation of biologic drugs.