Method and device for evaluating effect of shear force on stability of biological preparation or for optimizing production process of biological preparation

Abstract

The invention belongs to the field of biological medicine and fluid mechanics, and particularly relates to a method for rapidly calculating shear force in steps of filtering, sterilizing, filling and the like in production of fluids such as biological agents by utilizing fluid mechanics priori knowledge. And a reasonable experiment is designed to complete the evaluation of the influence of the shearing force on the fluid stability in a production scene by adopting a small amount of samples in a short time (such as 3 hours).

Description

Technical Field

[0001] The present invention belongs to the fields of biomedicine and fluid mechanics, and specifically involves using prior knowledge of fluid mechanics to quickly calculate the shear force in operations such as filtration sterilization and filling in the production of biological preparations, and designing reasonable experiments to complete the evaluation of the impact of shear force on the stability of biological preparations in production scenarios in a relatively short period of time (for example, within 3 hours) and using a small amount of sample. Background Art

[0002] In the production of biological preparations (such as pharmaceutical preparations of proteins, peptides, nucleic acids or their derivatives), sterilization filtration and filling operations are usually required. During this operation, biological preparations are often subjected to shear forces. Shear stress (σ) has been considered to be one of the important causes of aggregation and denaturation of proteins. Higher shear forces can destroy the multi-level structure of proteins, causing them to lose their original conformation and thus lose their biological activity, affecting their efficacy and safety. Therefore, in the process of laboratory preparation process development or in the production preparation engineering batch, the shear forces in operations such as sterilization filtration and filling are usually simulated to examine the protein stability during the preparation production process.

[0003] When evaluating the effect of shear stress on protein stability, traditional laboratory formulation process development or engineering batches often use the same scale as formal production to investigate shear stress. That is, using silicone tubing and filling needles with the same diameter as formal production, the protein stability changes before and after filtration and filling are evaluated at a pump speed slightly higher than the production pump speed, thereby investigating the stability of the protein during the production process. Although these methods can more accurately assess the effect of shear stress on proteins in actual production, they generally require a large sample volume (e.g., 250ml or more), a large number of manpower (>3 people), and a long preparation and experimental time (>12h). In addition, when using silicone tubing and filling needles of the same or similar dimensions for experiments, the same or slightly higher flow rate as production is often used for simulation. Although this can generate a shear stress greater than that in actual production, it fails to accurately simulate the shear stress distribution at different locations within the tubing. Moreover, because the length of the silicone tubing and the time during the experiment are difficult to accurately control according to actual production conditions, it is possible that although the laboratory shear stress is higher than the actual production, the experimental time is much shorter than the production time, and thus the changes in the protein under production conditions are not fully reflected. Therefore, it is of great practical significance to develop a laboratory method that can quickly, concisely, and cost-effectively evaluate the effect of shear stress on the stability of biologics in production scenarios.

[0004] The present application aims to provide a method for evaluating the effect of shear force on the stability of biological preparations. The method is rapid and concise, consumes little sample, saves time and effort, and has strong reliability and repeatability. It can accurately evaluate the effect of shear force on the stability of biological preparations in production scenarios to guide or optimize the production of biological preparations. Summary of the Invention

[0005] Summary of the Invention

[0006] In one aspect, the present invention provides a method for evaluating the effect of shear force on the stability of a biological agent in the production of a biological agent. The method is based on a fluid mechanics model, taking into account the effect of shear force magnitude, distribution and duration of action on the stability of a biological agent. It can use a smaller sample amount (e.g., within 10 ml) and complete the evaluation of the effect of shear force on the stability of a biological agent in a shorter time (e.g., within 1 hour, or even within a few minutes), and requires less manpower input, and has strong reliability and repeatability. The method can be carried out on a laboratory scale to accurately evaluate the effect of shear force on the stability of a biological agent in a production scenario (e.g., sterilizing filtration and / or filling operations) to guide the production of biological agents and provide a scientific basis for optimizing the production process of biological agents.

[0007] In another aspect, the present invention provides a method for optimizing a biologics production process.

[0008] In another aspect, the present invention provides a laboratory device for evaluating the effect of shear forces on the stability of a biologic during its production or for optimizing a biologic production process.

[0009] Various aspects of the present invention and its embodiments, features and advantages are described in detail below. Without departing from the concept of the present invention, other aspects and its embodiments, features and advantages not specifically described herein can be determined by those skilled in the art based on the description and subsequent claims. Detailed Description of the Invention

[0010] In one aspect, a method for evaluating the effect of shear stress on the stability of a biologic during its production is provided, comprising:

[0011] (a) Obtaining geometric parameters of the production passage and operating parameters of the production pump in the production scene;

[0012] (b) determining geometric parameters of the experimental path and operating parameters of the experimental pump in the laboratory scenario based on the production scenario parameters obtained in step (a);

[0013] (c) determining the required sample usage and evaluation time based on the production scenario parameters obtained in step (a) and the laboratory scenario parameters determined in step (b);

[0014] (d) running the evaluation time determined in step (c) in a laboratory scenario having the laboratory scenario parameters determined in step (b) using the pharmaceutical liquid or a simulated liquid thereof for preparing the biological agent in the sample usage amount determined in step (c); and

[0015] (e) detecting changes in the stability of the liquid obtained in step (d) to evaluate the effect of shear force on the stability of the biological preparation.

[0016] In another aspect, the present invention provides a method for optimizing a biologics production process, comprising:

[0017] (a) Setting the geometric parameters of the production path and the operating parameters of the production pump in the production scenario as required;

[0018] (b) determining the geometric parameters of the experimental path and the operating parameters of the experimental pump in the laboratory scenario based on the production scenario parameters set in step (a);

[0019] (c) determining the required sample usage and evaluation time based on the production scenario parameters set in step (a) and the laboratory scenario parameters determined in step (b);

[0020] (d) running the evaluation time determined in step (c) in a laboratory scenario having the laboratory scenario parameters determined in step (b) using the drug liquid or a simulated liquid thereof for preparing the biological agent in the sample usage amount determined in step (c);

[0021] (e) detecting a change in the stability of the liquid obtained in step (d) to evaluate the effect of shear force on the stability of the biological preparation; and

[0022] (f) When the detection of step (e) shows that the effect of shear force on the stability of the biological agent is acceptable, the geometric parameters of the production path and the operating parameters of the production pump in the corresponding production scenario are determined as the production process of the biological agent; when the detection of step (e) shows that the effect of shear force on the stability of the biological agent is unacceptable, the geometric parameters of the production path and the operating parameters of the production pump in the production scenario set in step (a) are adjusted, and steps (b) to (e) are repeated until the detection of step (e) shows that the effect of shear force on the stability of the biological agent is acceptable or desired.

[0023] Typically, the cross-section of a passage (including a production passage and a test passage) is circular or approximately circular.

[0024] In some embodiments, the production pathway comprises a production pipeline for conveying liquid (e.g., liquid for preparing the biologic) and an optional internal bore of a filling needle for filling the liquid. In some embodiments, the production pathway is a production pipeline for conveying liquid (e.g., liquid for preparing the biologic), such as during a filtration sterilization operation. In some embodiments, the production pathway comprises a production pipeline for conveying liquid (e.g., liquid for preparing the biologic) and an internal bore of a filling needle for filling the liquid, such as during a filling operation, or during a filtration sterilization and filling operation. In some embodiments, the production pipeline is substantially composed of silicone tubing.

[0025] In some embodiments, the production passage (e.g., the production tubing for transporting liquids and the optional internal channel of the filling needle) has more than one radius value. In some embodiments, the production passage or production tubing has a single radius value, i.e., is uniform in thickness. In other embodiments, the production passage has more than two radius values.

[0026] In some embodiments, the experimental pathway is a laboratory tubing for transporting or circulating a liquid (a liquid used to prepare the biologic or a simulated liquid thereof). In some embodiments, the experimental pathway or laboratory tubing is substantially composed of silicone tubing. Preferably, the experimental pathway or laboratory tubing has a uniform radius, i.e., is uniform in thickness.

[0027] In some embodiments, the geometric parameters of the production passage include the length L of each of the production passages of different radii. i and radius R i The working parameters of the production pump include the production pump speed PR of the production channels with different radii. i and the ratio of production flow to production pump speed k i , where i is any integer from 1 to m, and m is the number of radius values ​​in the production path.

[0028] In some embodiments, the geometric parameters of the experimental path include the length L of the experimental path. lab and radius R lab The working parameters of the experimental pump include the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab .

[0029] In some embodiments, the geometric parameters of the experimental passage and the operating parameters of the experimental pump are determined based on the following conditions:

[0030] (i) Experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed klab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than the production pump speed PR of each production channel with different radius i and the ratio of production flow to production pump speed k i The product of divided by the radius R of the production channel i The quotient of the cube of (PR i ·k i ) / R i 3 The maximum value in

[0031]

[0032] (ii) Optionally, the length L of the experimental path lab With radius R lab The ratio L lab / R lab is the length L of the production channels with different radii i With radius R i The ratio L i / R i The sum of about 0.8 to about 10 times of;

[0033] wherein i is any integer from 1 to m, and m is the number of radius values ​​in the production path; and

[0034] Among them, the length of the experimental path L lab It refers to the length of the path that the liquid flows through in the laboratory background device. It can be understood that when the liquid circulates n times in the laboratory device, the length of the experimental path L lab It is numerically equal to the product of the length of the experimental silicone tube and the number of cycles n.

[0035] In some embodiments, for (i), the experimental pump rate PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 is the production pump speed PR of production channels with different radii i and the ratio of production flow to production pump speed k i The product of divided by the radius R of the production channel i The quotient of the cube of (PR i ·ki ) / R i 3 1 to about 100 times, for example, about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times the maximum value in .

[0036] In some embodiments, for (ii), the length L of the experimental path is lab With radius R lab The ratio L lab / R lab is the length L of the production channels with different radii i With radius R i The ratio L i / R i The sum of The amount of the active ingredient is about 0.8 to about 5 times, for example, about 0.8 to about 2 times, about 1 to about 1.5 times, or about 1 to about 1.2 times.

[0037] In some embodiments, the length L of each of the production passages of different radii is based on i and radius R i , the radius R of the experimental path lab , the experimental pump speed PR lab and the ratio k of the experimental flow rate to the experimental pump speed lab To determine the evaluation time t lab .

[0038] In some embodiments, the evaluation time t lab yes The amount of the present invention is about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 time.

[0039] In some embodiments, based on the length L of the experimental pathway lab and radius R lab The amount of sample used for the biological agent is determined based on the number of cycles n of the sample in the device in the laboratory scenario, wherein n is any integer or decimal greater than or equal to 1. In some embodiments, the minimum amount of sample used for the biological agent is (πR lab 2 L lab ) / n, where n is any integer or decimal greater than or equal to 1. Preferably, n is an integer greater than or equal to 1. It is understood that when n is a decimal greater than or equal to 1, it means that the sample or liquid completes an integer number of complete cycles in the laboratory device and then flows through an additional part of the pipeline but does not complete a complete cycle. It is also understood that when n is 1, the minimum sample usage of the biological agent is πR lab2 L lab .

[0040] When implementing the method of the present invention, the actual experimental sample usage amount used may be equal to or slightly higher than the minimum sample usage amount. In some embodiments, the actual sample usage amount of the biological preparation may also be slightly lower than or lower than the minimum sample usage amount.

[0041] Filtration sterilization operation

[0042] The evaluation method of the present invention can be used to evaluate the effect of shear force on the stability of a biological preparation during the filtration sterilization operation of the biological preparation. The optimization method of the present invention can be used to optimize the production process of the filtration sterilization operation of the biological preparation.

[0043] In some embodiments, the production pathway is a production line used to transport liquids during filtration and sterilization operations in the production of biologics, and the experimental pathway is an experimental line used to transport or circulate liquids.

[0044] In some embodiments, the production passage or the production pipeline for transporting liquid is uniform in thickness, that is, has the same radius. In some embodiments, the experimental passage is uniform in thickness, that is, has the same radius.

[0045] In some embodiments, the geometric parameters of the production passage include the length L of the production passage. filt and radius R filt The working parameters of the production pump include the production pump speed PR filt and the ratio of production flow to production pump speed k filt .

[0046] In some embodiments, the geometric parameters of the experimental path include the length L of the experimental path. lab and radius R lab The working parameters of the experimental pump include the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab .

[0047] In some embodiments, the geometric parameters of the experimental passage and the operating parameters of the experimental pump are determined based on the following conditions:

[0048] (i) Experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than production pump speed PRfilt and the ratio of production flow to production pump speed k filt The product of divided by the radius R of the production channel filt The quotient of the cube of (PR filt ·k filt ) / R filt 3 ,Right now

[0049] and

[0050] (ii) Optionally, the length L of the experimental path lab With radius R lab The ratio L lab / R lab is the length of the production channel L filt With radius R filt The ratio L filt / R filt about 0.8 to about 10 times of;

[0051] Among them, the length of the experimental path L lab It refers to the length of the path that the liquid flows through in the laboratory background device. It can be understood that when the liquid circulates n times in the laboratory device, the length of the experimental path L lab It is numerically equal to the product of the length of the experimental silicone tube and the number of cycles n.

[0052] In some embodiments, for (i), the experimental pump rate PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 is the production pump speed PR filt and the ratio of production flow to production pump speed k filt The product of divided by the radius R of the production channel filt The quotient of the cube of (PR filt ·k filt ) / R filt 3 The amount of the active ingredient is about 1 to about 100 times, for example, about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times.

[0053] In some embodiments, for (ii), the length L of the experimental path is lab With radius R lab The ratio L lab / R lab is the length of the production channel L filtWith radius R filt The ratio L filt / R filt The amount of the active ingredient is about 0.8 to about 5 times, for example, about 0.8 to about 2 times, about 1 to about 1.5 times, or about 1 to about 1.2 times.

[0054] In some embodiments, based on the length L of the production path filt and radius R filt , the radius R of the experimental path lab , the experimental pump speed PR lab and the ratio k of the experimental flow rate to the experimental pump speed lab To determine the evaluation time t lab .

[0055] In some embodiments, the evaluation time t lab yes The amount of the present invention is about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 time.

[0056] In some embodiments, based on the length L of the experimental pathway lab and radius R lab The amount of sample used for the biological agent is determined based on the number of cycles n of the sample in the device in the laboratory scenario, wherein n is any integer or decimal greater than or equal to 1. In some embodiments, the minimum amount of sample used for the biological agent is (πR lab 2 L lab ) / n, where n is any integer or decimal greater than or equal to 1. Preferably, n is an integer greater than or equal to 1. It is understood that when n is a decimal greater than or equal to 1, it means that the sample or liquid completes an integer number of complete cycles in the laboratory device and then flows through an additional part of the pipeline but does not complete a complete cycle. It is also understood that when n is 1, the minimum sample usage of the biological agent is πR lab 2 L lab .

[0057] When implementing the method of the present invention, the actual experimental sample usage amount used may be equal to or slightly higher than the minimum sample usage amount. In some embodiments, the actual sample usage amount of the biological preparation may also be slightly lower than or lower than the minimum sample usage amount.

[0058] In some embodiments, the production line for transporting the liquid can have different radii, for example, at least two radius values. In some embodiments, when the production line has at least two radius values, the liquid used to prepare the biological agent can be cycled through steps (a) to (d), with each cycle being for each production line having a different radius, and then proceeding to step (e). It will be understood that in multiple cycles of steps (a) to (d), step (e) is optionally performed after step (d), and if it is found that the shear force has caused the biological agent to be unstable, the subsequent cycle of steps (a) to (d) can be terminated.

[0059] Filling operation

[0060] The evaluation method of the present invention can also be used to evaluate the effect of shear force on the stability of a biological preparation during the filling operation of the biological preparation. The optimization method of the present invention can also be used to optimize the production process of the filling operation of the biological preparation.

[0061] In some embodiments, the production path includes a production line for conveying liquid and an internal channel of a filling needle for filling liquid during a filling operation of a biological preparation production, and the experimental path is an experimental line for conveying or circulating liquid.

[0062] In some embodiments, the production pipeline for transporting liquid is uniform in thickness, that is, has the same radius. In some embodiments, the experimental pipeline is uniform in thickness, that is, has the same radius.

[0063] In some embodiments, the geometric parameters of the production path include the length L of the production line. fill and radius R fill And the length of the filling needle L needle and radius R needle The working parameters of the production pump include the production pump speed PR fill and the ratio of production flow to production pump speed k fill .

[0064] In some embodiments, the geometric parameters of the experimental path include the length L of the experimental path. lab and radius R lab The working parameters of the experimental pump include the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab .

[0065] In some embodiments, the geometric parameters of the experimental passage and the operating parameters of the experimental pump are determined based on the following conditions:

[0066] (i) Experimental pump speed PR laband the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill The product of divided by the radius R of the production pipeline fill The quotient of the cube of (PR fill ·k fill ) / R fill 3 and filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·k fill ) / R fill 3 The maximum value in

[0067] and

[0068] (ii) Optionally, the length L of the experimental path lab With radius R lab The ratio L lab / R lab is the length of the production pipeline L fill and radius R fill The ratio of the filling needle length L needle and radius R needle The sum of the ratios L fill / R fill +L needle / R needle about 0.8 to about 10 times of;

[0069] Among them, the length of the experimental path L lab It refers to the length of the path that the liquid flows through in the laboratory background device. It can be understood that when the liquid circulates n times in the laboratory device, the length of the experimental path L lab It is numerically equal to the product of the length of the experimental silicone tube and the number of cycles n.

[0070] In some embodiments, for (i), the experimental pump rate PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / Rlab 3 Is the filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill The product of divided by the radius R of the production pipeline fill The quotient of the cube of (PR fill ·k fill ) / R fill 3 Filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·k fill ) / R needle 3 1 to about 100 times, for example, about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times the maximum value in .

[0071] In some embodiments, for (ii), the length L of the experimental path is lab With radius R lab The ratio L lab / R lab is the length L of the production pipeline fill and radius R fill The ratio of the filling needle length L needle and radius R needle The sum of the ratios L fill / R fill +L needle / R needle The amount of the active ingredient is about 0.8 to about 5 times, for example, about 0.8 to about 2 times, about 1 to about 1.5 times, or about 1 to about 1.2 times.

[0072] Usually, in the production filling equipment, the radius R of the production pipeline fill Is larger than the filling needle radius R needle Therefore, the above condition (i) can be simplified to: experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·kfill ) / R fill 3 ,Right now

[0073]

[0074] In some embodiments, for simplified condition (i), the experimental pump rate PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Is the filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·k fill ) / R needle 3 The amount of the active ingredient is about 1 to about 100 times, for example, about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times.

[0075] In some embodiments, based on the geometric parameters of the production path (including the length L of the production line), fill and radius R fill And the length of the filling needle L needle and radius R needle ), the radius R of the experimental path lab , the experimental pump speed PR lab and the ratio k of the experimental flow rate to the experimental pump speed lab To determine the evaluation time t lab .

[0076] In some embodiments, the evaluation time t lab yes The amount of the present invention is about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 time.

[0077] In some embodiments, based on the length L of the experimental pathway lab and radius R lab And the sample usage of the biological agent is determined based on the number of cycles n of the sample in the device of the laboratory scene, wherein n is any integer or decimal greater than or equal to 1. In some embodiments, the minimum sample usage of the biological agent is (πR lab 2 Llab ) / n, where n is any integer or decimal greater than or equal to 1. Preferably, n is an integer greater than or equal to 1. It is understood that when n is a decimal greater than or equal to 1, it means that the sample or liquid completes an integer number of complete cycles in the laboratory device and then flows through an additional part of the pipeline but does not complete a complete cycle. It is also understood that when n is 1, the minimum sample usage of the biological agent is πR lab 2 L lab .

[0078] When implementing the method of the present invention, the actual experimental sample usage amount used may be equal to or slightly higher than the minimum sample usage amount. In some embodiments, the actual sample usage amount of the biological preparation may also be slightly lower than or lower than the minimum sample usage amount.

[0079] Furthermore, the evaluation method of the present invention can be used to evaluate the effect of shear force on the stability of a biological preparation during the filtration sterilization and filling operations of the biological preparation. The optimization method of the present invention can be used to optimize the production process of the filtration sterilization and filling operations of the biological preparation.

[0080] In some embodiments, more than one pump may be connected in parallel in a laboratory setting to provide greater shear force. In this case, the experimental pump rate PR in condition (i) described herein is lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 represents the sum of the quotients of each pump, that is, and the evaluation time t described in this paper lab yes about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 times, wherein PR lab,i is the pump speed of the i-th pump in the laboratory, k lab,i is the pump flow / pump speed of the ith pump in the laboratory, num_pump is the number of pumps in the laboratory, L i and R i are the length and radius of production channels with different radii, and m is the number of radius values ​​in the production channel.

[0081] Shear force is a mechanical stress that is mainly caused by the friction between fluid particles caused by the viscosity of the fluid. Liquids used to prepare biological preparations are exposed to shear forces during filtration, sterilization and filling operations. Shear force is known to be an important cause of aggregation and denaturation of proteins, etc. In traditional laboratory evaluation methods, the same or slightly higher flow rate as production is usually used to simulate the shear force effects of actual production, but the shear force is different at different (cross-sectional) positions in the production pipeline. Traditional laboratory evaluation methods fail to accurately simulate the shear force distribution in the pipeline. The present invention proposes a scheme for shear force evaluation during sterilization filtration and / or filling based on a fluid mechanics model (see Figure 1 Specifically, the shear force (σ) at different locations inside the production pipeline during the filtration sterilization operation was calculated using formula (1). filt is the shear force at the calculated position in the production pipeline, Q filt is the sample flow rate in production (m 3 / s), μ is the sample viscosity, R filt is the radius of the production pipeline, and r is the distance from the calculated position to the center of the pipeline cross section):

[0082]

[0083] From formula (1), we can see that the shear force on the sample near the tube wall is the largest (see formula (2), σ filt,max The shear force in the center of the pipeline is 0 (r = 0), and the shear force distribution from the center of the pipeline to the pipe wall is linear (see Figure 2 ).

[0084]

[0085] The average shear stress (σ filt,avg ) is as follows (PR filt is the production pump speed, k filt is the production pump flow / pump speed):

[0086]

[0087] In the laboratory evaluation method, the shear force must be greater than or equal to the shear force in the actual production of the preparation to simulate the worst condition, that is, (σ lab,avg is the average shear force in the laboratory pipeline):

[0088] σ lab,avg ≥σ filt,avg Eq.(4)

[0089] Therefore, the laboratory evaluation method needs to satisfy formula (5) (where PR lab is the pumping speed of the laboratory pump, klab is the laboratory pump flow / pump speed, R lab is the laboratory pipeline radius):

[0090]

[0091] Right now

[0092] In addition, the effect of shear force on biological agents should take into account the duration of action. The production process time can be calculated by formula (7) (where L fill is the length of production pipeline):

[0093]

[0094] The effect of shear stress on biological agents during production is defined according to formula (8):

[0095]

[0096] Therefore, from formula (8), it can be seen that the laboratory evaluation method must also satisfy formula (9) (where L lab is the length of the experimental path, R lab is the radius of the experimental path):

[0097]

[0098] Right now

[0099] At the same time, in order to save costs and samples, the laboratory evaluation method can consider connecting a thinner and shorter pipe and then circulating it. Accordingly, it is necessary to consider controlling the experimental time to meet formula (11):

[0100]

[0101] Right now

[0102] At this time, the minimum sample requirement is

[0103] DS_amount min =πR lab 2 L lab Eq.(13)

[0104] Furthermore, the sample can be circulated in the laboratory device n times (n is any integer or decimal greater than or equal to 1), then the length of the experimental path L lab It is numerically equal to the length of the experimental silicone tube (L lab-硅胶管) multiplied by the number of cycles n, i.e., the length of the path that the sample travels in the laboratory device. In other words, the sample can be circulated n times in the laboratory device to reach a length of n·L. lab-硅胶管 The same effect as the experimental silicone tube. At this time, the minimum sample requirement is

[0105]

[0106] In summary, the experimental design can be found in Figure 3 .

[0107] Similarly, during the filling process, the average shear force on the sample in the filling production pipeline and filling needle can be calculated according to formulas (15) and (16) (σ fill,avg is the average shear force in the filling production pipeline, σ needle,avg is the average shear force in the filling needle, Q fill is the sample flow rate during filling (m 3 / s), R fill is the radius of the filling production pipeline, R needle is the filling needle radius, PR fill is the pump speed, k fill is flow rate / pump speed):

[0108]

[0109]

[0110] In the laboratory evaluation method, the average shear force must be greater than or equal to the maximum average shear force in actual production to simulate the worst conditions, that is, (σ lab,avg is the average shear force in the laboratory pipeline):

[0111] σ lab,avg ≥max(σ fill,avg ,σ needle,avg ) Eq.(17)

[0112] Therefore, the laboratory evaluation method needs to satisfy formula (18) (where PR lab is the pump speed of the experimental pump, k lab is the experimental pump flow / pump speed, R lab is the laboratory pipeline radius):

[0113]

[0114] Usually, the radius R of the filling pipe fill Greater than the radius R of the inner channel of the filling needle needle , so the formula can be expressed as:

[0115]

[0116] The time the sample is subjected to shear force is (L fill is the length of the filling production pipeline, L needle is the filling needle length):

[0117]

[0118]

[0119] The shear force experienced during the filling process is

[0120]

[0121] At this time, it is necessary to satisfy formula (23) (where L lab is the length of the experimental path, R lab is the radius of the experimental path):

[0122]

[0123] In order to save costs and samples, laboratory evaluation methods can consider connecting thinner and shorter pipes for circulation. Accordingly, in this case, it is necessary to consider controlling the experimental time to meet the following formula:

[0124]

[0125] Right now

[0126] Similarly, according to the length L of the experimental path lab and radius R lab The minimum sample requirement is

[0127] DS_amount min =πR lab 2 L lab Eq.(13)

[0128] Furthermore, the sample can be circulated n times in the laboratory device to achieve the same length as n·L. lab-硅胶管 The same effect as the experimental silicone tube. At this time, the minimum sample requirement is

[0129]

[0130] Wherein, n is any integer or decimal greater than or equal to 1, representing the number of times the sample circulates in the device in the laboratory scenario.

[0131] The evaluation and optimization methods of the present invention are also applicable to production scenarios where there are multiple different pipe radii in the pipeline. In particular, the evaluation and optimization methods of the present invention can be performed using a set of laboratory equipment or a set of laboratory parameters, which is convenient in operation, saves time and effort, and is cost-effective. For example, the average shear force to which the sample is subjected during the filtration, sterilization and filling process can be calculated according to the following formula:

[0132]

[0133]

[0134]

[0135] Formulas (1-1), (1-2) and (1-3) are for sterilization filtration (filt), filling (fill) and filling needle (needle), respectively, where σ 1,avg , σ 2,avg and σ 3,avg They represent the average shear force in the sterilizing and filtration production pipeline, the average shear force in the filling production pipeline and the average shear force in the filling needle respectively, Q is the sample flow rate, R1, R2 and R3 are the pipeline or channel radius respectively, PR1, PR2 and PR3 are the pump speeds respectively, k1, k2 and k3 are the pump flow rate / pump speed respectively, among which PR2 is consistent with PR3, and k2 is consistent with k3.

[0136] Furthermore, considering that different pipeline radii may exist during sterilization filtration and filling, the average shear force in channels with different radii can be calculated as follows:

[0137]

[0138] Wherein, i is a positive integer from 1 to m, and m is the number of radius values ​​in the production path.

[0139] If the laboratory setup uses one silicone tubing radius, then

[0140] σ lab,avg ≥max(σ 1,avg ,σ 2,avg ,σ 3,avg ,…,σ m,avg ) Eq.(2-1)

[0141] Right now,

[0142] At the same time, the total time for sterilization, filtration and filling is:

[0143]

[0144] The total effect of shear stress on biologics during production is:

[0145]

[0146] Therefore, the laboratory scenario can be designed as follows:

[0147] The experimental time can be controlled as follows:

[0148]

[0149] Right now,

[0150] Similar to the above, the minimum sample requirement is calculated as follows:

[0151]

[0152] Where n is any integer or decimal greater than or equal to 1, representing the number of times the sample circulates in the laboratory device.

[0153] In some embodiments, multiple pumps may be included in a laboratory setting. In this case, the pumps are typically connected in parallel to provide sufficient shear force. In this case, the laboratory shear force can be calculated using the following formula:

[0154]

[0155] PR lab,i is the pump speed of the i-th pump in the laboratory, k lab,i is the pump flow / pump speed of the i-th pump in the laboratory, and num_pump is the number of pumps in the laboratory.

[0156] The laboratory process needs to meet the shear force greater than the shear force in the production sterilization filtration and filling process, that is:

[0157]

[0158]

[0159]

[0160] Furthermore, combined with Eq. (2-2), we get the following formula:

[0161]

[0162] PR lab,i ·k lab,i The variables are eliminated, so the formula for the effect of shear force on protein preparations in production does not change. Therefore, similarly, the laboratory scenario can also be designed to meet the following formula:

[0163] The experiment run time can be set to:

[0164]

[0165] Similar to the above, the minimum sample requirement is calculated as follows:

[0166]

[0167] Where n is any integer or decimal greater than or equal to 1, representing the number of times the sample circulates in the laboratory pathway.

[0168] In this article, the meaning of the symbols in each formula can be understood in a universal sense. Therefore, even if the meaning of certain symbols is not given directly, those skilled in the art can clearly determine them according to the context. For example, in this article, σ is the shear force, Q is the sample flow rate, μ is the sample viscosity, L is the passage or pore length, R is the passage or pore radius, PR is the pump speed, k is the ratio of pump flow / pump speed, and t is the experimental evaluation time; the subscript filt indicates filtration sterilization, fill indicates the production pipeline for filling, needle indicates the filling needle, lab indicates the laboratory scene, and avg indicates the average value. Therefore, it can be understood that σ filt represents the shear force of filtration sterilization, σ filt,avg Represents the average shear force during filtration sterilization. Other symbols can be understood accordingly.

[0169] Biological preparations and their stability testing

[0170] In some embodiments, the biologic is susceptible to aggregation or denaturation under shear forces. In some embodiments, the biopharmaceutical molecules of the biologic are susceptible to aggregation or denaturation under shear forces. In some embodiments, the biopharmaceutical molecules of the biologic are selected from one or more of proteins, polypeptides, nucleic acids, or any derivatives thereof (e.g., fusions or conjugates), such as, but not limited to, antibodies (e.g., monoclonal antibodies or polyclonal antibodies) or antigen-binding fragments thereof, fusion proteins, biopharmaceutical conjugates (XDCs) such as antibody-drug conjugates (ADCs), polypeptides, and small nucleic acid drugs.

[0171] In some embodiments, the biologic can be a protein or conjugate preparation, a polypeptide or conjugate preparation, a nucleic acid or conjugate preparation. In some embodiments, the biologic is an antibody (such as a monoclonal antibody or polyclonal antibody) or its antigen-binding fragment, a fusion protein, a bioconjugate drug (XDC) such as an antibody-drug conjugate (ADC), a polypeptide, or a small nucleic acid drug. In some preferred embodiments, the biologic is a pharmaceutical preparation of an antibody or its antigen-binding fragment, such as a monoclonal antibody or polyclonal antibody or its antigen-binding fragment. In other preferred embodiments, the biologic is a bioconjugate drug (XDC) preparation, such as an antibody-drug conjugate (ADC) preparation.

[0172] In some embodiments, the biologic is an injectable biologic. In some embodiments, the biologic is a liquid formulation. In some embodiments, the biologic is a solution formulation, such as an aqueous solution formulation. In some embodiments, the biologic is in the form of a spray-dried powder or a freeze-dried powder.

[0173] In some embodiments, the liquid for preparing the biological preparation is a solution of the biopharmaceutical molecule comprising the biological preparation, such as an aqueous solution, which optionally includes one or more excipients. In some embodiments, the liquid for preparing the biological preparation is a liquid comprising the biopharmaceutical molecule of the biological preparation and all excipients. In other embodiments, the liquid for preparing the biological preparation is a liquid comprising a part of the excipients comprised by the biopharmaceutical molecule of the biological preparation and the biological excipient. Excipient is that those skilled in the art can routinely determine, and its non-limiting example includes buffer, surfactant, osmotic pressure regulator, stabilizer (such as sugar and polyol, complexing agent, cyclodextrin, etc.), solubilizer, cosolvent, antioxidant, adsorption inhibitor, preservative, local pain reliever, etc. In some embodiments, the liquid for preparing the biological preparation can additionally include one or more small molecule chemical drugs. In some embodiments, the liquid for preparing the biological preparation is the drug stock solution of the biological preparation.

[0174] In some embodiments, the liquid used in the methods of the present invention is a simulant of the liquid used to prepare the biological agent.

[0175] In some embodiments, the liquid for preparing a biologic, or a liquid simulating the same, has a viscosity of less than 1300 cp, such as less than 1000 cp, less than 500 cp, less than 100 cp, less than 50 cp, less than 25 cp, such as about 0.1-20 cp, 0.1-10 cp, or 0.1-5 cp. In some embodiments, the liquid for preparing a biologic, or a liquid simulating the same, is a Newtonian fluid. In some embodiments, the liquid for preparing a biologic, or a liquid simulating the same, is a non-Newtonian fluid.

[0176] In the methods of the present invention, for step (e), any item of biopharmaceutical stability can be used to assess the effect of shear stress on the stability of the biopharmaceutical. In some embodiments, the assessment is achieved by detecting changes in the physical stability and / or chemical stability of the biopharmaceutical molecule. In some embodiments, the assessment is achieved by detecting changes in the physical stability and / or chemical stability of the biopharmaceutical molecule before and after performing the methods of the present invention. In some embodiments, instability manifests as one or more of aggregation, denaturation, adsorption, precipitation, hydrolysis, degradation, and / or oxidation.

[0177] In some embodiments, non-limiting examples of biopharmaceutical stability test items include, but are not limited to, one or more of the following: appearance, including clarity or turbidity, color, visible particles; subvisible particles; pH; biopharmaceutical molecule concentration; biopharmaceutical molecule molecular weight; viscosity; biological activity; purity (e.g., SEC, CE-SDS); degradation product content; charge variants (iCIEF); for antibody-drug conjugates, average Dar value and / or free toxin (e.g., SEC, RP-HPLC, HIC-HPLC, etc.). It will be understood that other test items not specifically mentioned herein may also be used in the methods of the present invention. Test items and methods for biopharmaceutical molecule stability can be determined by those skilled in the art.

[0178] Laboratory equipment

[0179] On the other hand, the present invention provides a laboratory device for the method of the present invention for evaluating the effect of shear force on the stability of a biological agent in the production of a biological agent or the method of the present invention for optimizing the production process of a biological agent, the device comprising an experimental pipeline 1 and an experimental pump 2 that are liquid-connected to each other, and having a liquid inlet 3 and a liquid outlet 4 on the experimental pipeline, wherein the experimental pipeline 1 has geometric parameters determined according to the method of the present invention, and the experimental pump 2 is configured to have working parameters determined according to the method of the present invention. Figure 4 A schematic diagram of a laboratory setup that can be used for the evaluation method or optimization method of the present invention is shown. Figure 5A schematic diagram of another laboratory device that can be used for the evaluation method or optimization method of the present invention is shown, wherein the liquid can circulate n times in the pipeline, where n is any integer or decimal greater than or equal to 1.

[0180] In some embodiments, the cross-section of the experimental tubing is circular or approximately circular. In some embodiments, the experimental tubing is made of the same material as the production tubing. In some embodiments, the experimental tubing is essentially composed of silicone tubing.

[0181] In some embodiments, the experimental pump is a peristaltic pump. In some embodiments, the experimental pump is a peristaltic hose pump, which delivers liquid by squeezing and releasing an elastic hose, such as a silicone tube.

[0182] In some embodiments, the laboratory device of the present invention may optionally include one or more elements selected from the following: a liquid supply element, a liquid receiving element, a metering element, a timing element, a switching element, a parameter monitoring and / or control element (e.g., a pump speed monitoring and / or control element, a flow monitoring and / or control element, a shear force monitoring and / or control element, a viscosity monitoring and / or control element, a temperature monitoring and / or control element), a cycle number control and / or recording element, and a stability detection element.

[0183] In some embodiments, the laboratory device of the present invention can optionally include one or more liquid supply elements. In some embodiments, the liquid supply element includes a storage container. In some embodiments, the storage container can be selected from a tank, bottle, bucket, bag or other suitable container. In some embodiments, the liquid supply element is equipped with a liquid, such as a liquid for preparing the biological agent. In some embodiments, the liquid supply element is connected to a liquid inlet 3.

[0184] In some embodiments, the laboratory device of the present invention can optionally include one or more liquid receiving elements. In some embodiments, the liquid receiving element comprises a receiving container. In some embodiments, the receiving container can be selected from a plastic bottle, an ampoule, a vial, a test tube, a flask, a beaker, a bag, a bucket, a cylinder, a syringe, etc. In some embodiments, the liquid receiving element is connected to the liquid outlet 4.

[0185] In some embodiments, the laboratory device of the present invention can optionally include a metering element. In some embodiments, the metering element is a metering valve. In some embodiments, the metering valve can provide a metering volume of about 0.1 ml to about 50 ml, for example, about 1-40 ml, about 1-30 ml, about 1-20 ml, about 1-15 ml, about 1-10 ml, or about 1-5 ml. In some embodiments, the metering element is placed at the liquid inlet 3.

[0186] In some embodiments, the laboratory device of the present invention can optionally include a timing element. In some embodiments, the timing element is used to measure and / or control the running time of the liquid in the laboratory device. In some embodiments, the timing device is an automatic timer or stopwatch.

[0187] In some embodiments, the laboratory device of the present invention may optionally include a switch element. Preferably, the switch element is a timer switch element. When the liquid running in the laboratory pipeline reaches a determined evaluation time, the timer switch element can automatically stop the liquid from running.

[0188] In some embodiments, the laboratory device of the present invention may also optionally include parameter monitoring and / or control elements. In some embodiments, the parameters include pump speed, flow rate, shear force, liquid viscosity, temperature, etc. In some embodiments, the parameter monitoring and / or control elements include pump speed monitoring and / or control elements, flow rate monitoring and / or control elements, shear force monitoring and / or control elements, viscosity monitoring and / or control elements, temperature monitoring and / or control elements, etc.

[0189] In some embodiments, the laboratory-device of the present invention may optionally include a cycle time control and / or recording element.

[0190] In some embodiments, the laboratory device of the present invention may further optionally include a stability detection element, which is connected to the liquid outlet 4 so as to directly receive the liquid flowing out of the laboratory device for detection.

[0191] In some embodiments, the pipeline of the laboratory device of the present invention can be divided into multiple pipeline sections, which connect the laboratory pump and optional parameter monitoring and / or control elements, optional cycle number control and / or recording elements, etc. to each other so that liquid can flow therein once (see Figure 4 ) or loop n times (see Figure 5 ) passage.

[0192] Compared with the existing technology, the method and laboratory device of the present invention can accurately reflect the effect of shear force on liquid stability in production scenarios. The method is fast and concise, the device is simple, the sample consumption is small, it saves time and labor, and has strong reliability and repeatability.

[0193] It is understood that the various aspects and their embodiments, features, and advantages described herein are equally applicable to the other aspects of the present invention, including aspects not directly described, and their various embodiments, features, and advantages. Other aspects and their embodiments, features, and advantages not specifically described herein are readily ascertainable by those skilled in the art from the present disclosure and the claims that follow without departing from the scope of the present invention.

[0194] definition

[0195] It should be understood that the terms used herein are merely for the purpose of describing embodiments of the present invention and are not intended to limit the scope of the present invention. The terms used in this application have the meanings defined below, unless otherwise specified or the context clearly contradicts. Terms not clearly defined in this application have the meanings generally understood by those skilled in the art.

[0196] As used in this application, the terms "a," "an," "the" and similar referents are to be construed to cover both the singular and the plural, unless otherwise indicated or clearly contradicted by context.

[0197] The term "about" when used in conjunction with a numerical value is meant to encompass numerical values ​​within a range having a lower limit that is 5%, 4%, 3%, 2%, or 1% less than the specified numerical value and an upper limit that is 5%, 4%, 3%, 2%, or 1% greater than the specified numerical value. The term "about" when used in conjunction with a numerical range is meant to encompass a range whose lower limit is 5%, 4%, 3%, 2%, or 1% less than the lower limit of the numerical range and whose upper limit is 5%, 4%, 3%, 2%, or 1% greater than the upper limit of the numerical range.

[0198] As used herein, the term "and / or" means any one of the optional items or two or more or all of the optional items. For example, the expression "A and / or B" includes the situations of A, B, and A+B.

[0199] As used herein, the term "comprising" or "including" means including the stated elements, integers or steps, but not excluding any other elements, integers or steps. In this document, when the term "comprising" or "including" is used, unless otherwise indicated, the situation consisting essentially of the stated elements, integers or steps and the situation consisting of the stated elements, integers or steps are also encompassed.

[0200] The term "consisting essentially of" means that other than the defined components, no other components are present in significant amounts, e.g., the amount of said other components is less than 10% w / w, preferably less than 5% w / w, more preferably less than 3% w / w, most preferably less than 1% w / w. The term "w / w" means weight ratio.

[0201] The term "pathway" refers to the path through which liquid flows within a device, encompassing both the tubing used to transport liquid and the internal bore of a filling needle used to fill liquid. Specifically, the term "production pathway" refers to the path through which liquid flows within a device in a production setting, and the term "experimental pathway" refers to the path through which liquid flows within a device in a laboratory setting.

[0202] When the liquid circulates n times in the laboratory device, the "length" (L lab ) refers to the length of the path that the liquid flows through in the laboratory device, which is numerically equal to the length of the experimental pipeline such as silicone tube (L lab-管路 or L lab-硅胶管 ) multiplied by the number of loops n.

[0203] The term "pipeline" refers to a pipe for conveying liquid, such as a silicone tube, or a pipe made of other materials. Preferably, the cross section of the pipe is circular or approximately circular.

[0204] The phrase "consisting essentially of silicone tubing" means that the tubing is silicone tubing, but does not preclude the presence of connecting components between tubing segments or between the tubing and other components in the device. These connecting components may be made of materials other than silicone. The length of these connecting components is included in the length of the production or experimental pathways.

[0205] The term "pipe segment" refers to the length of a pipeline between two adjacent connection points (e.g., connection points to a component or device). A pipeline can include one or more pipe segments, which can have the same or different lengths and / or the same or different radii. Typically, the one or more pipe segments have the same radius.

[0206] The term "inner bore" of a filling needle refers to the hollow, lumen-like passage of the filling needle, which is used to deliver liquid into a liquid-receiving container, such as a vial, ampoule, or vial.

[0207] The term "pump" refers to a device used to move a fluid (e.g., a liquid) through a circuit. The term "production pump" refers to a pump used in production settings, while the term "laboratory pump" refers to a pump used in laboratory settings. Production pumps include "filling pumps," which are devices used to fill liquids into receiving containers during the filling process.

[0208] The term "peristaltic pump" refers to a peristaltic hose pump, which transports liquids by squeezing and releasing a flexible hose, such as silicone tubing. In a peristaltic pump, the liquid only comes into contact with the hose, not the pump itself. In this context, the length of the tubing includes the length of the pump tubing within the peristaltic pump; that is, the pump tubing is considered part of the tubing.

[0209] The term "biologic," also referred to as "biotech drug formulation," refers to a pharmaceutical formulation comprising a biopharmaceutical molecule, optionally comprising one or more excipients. The biologic can be in liquid or powder form (e.g., spray-dried powder or freeze-dried powder). Preferably, the biologic is an injectable biologic, for example, administered by subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intraperitoneal, abdominal, spinal, joint, intracardiac, thoracic, intrathecal, or intravitreal route.

[0210] The term "biopharmaceutical molecule" refers to a biotechnology drug molecule that can produce a therapeutic or preventive effect, for example, a protein, a polypeptide, a nucleic acid, or any derivative thereof (e.g., a fusion or conjugate). For example, the biopharmaceutical molecule can be selected from an antibody (e.g., a monoclonal antibody or a polyclonal antibody) or an antigen-binding fragment thereof, a fusion protein, a biopharmaceutical conjugate (XDC) such as an antibody-drug conjugate (ADC), a polypeptide, or a small nucleic acid drug. The biopharmaceutical molecule is susceptible to aggregation or denaturation under the action of shear forces.

[0211] The term "protein" refers to a biological macromolecule composed of amino acids linked by peptide bonds to form a polypeptide chain, which further folds and coils to form a specific spatial structure. Protein molecules are generally prone to aggregation or denaturation under the influence of shear forces.

[0212] As used herein, the term "antibody" is used in the broadest sense and encompasses a variety of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, as long as they exhibit the desired antigen-binding activity. A complete antibody will generally comprise at least two full-length heavy chains and two full-length light chains, but may comprise fewer chains in certain circumstances, e.g., antibodies naturally occurring in camels may comprise only heavy chains.

[0213] The term "antigen-binding fragment" means a molecule different from an intact antibody, which comprises a portion of an intact antibody and is capable of binding to the antigenic epitope to which the intact antibody binds. Examples of antigen-binding fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; single-chain antibodies (e.g., scFv); single-domain antibodies; camelid antibodies (heavy-chain antibodies). Antigen-binding fragments can be prepared by recombinant DNA technology, or by enzymatic or chemical cleavage of intact antibodies. Unless otherwise specified herein or clearly contradicted by the context, the term "antibody" herein is equivalent to "antibody or antigen-binding fragment thereof."

[0214] The term "antibody-drug conjugate" or "ADC" refers to a substance obtained by coupling a small molecule drug toxin (payload) to an antibody or its antigen-binding fragment (responsible for targeting function and sometimes also having biological activity) via a linker. As used herein, the term "antibody-drug conjugate" or "ADC" includes pharmaceutically acceptable salts or esters, solvates, isotopically labeled substances (such as deuterated derivatives), stereoisomers, tautomers, and other equivalent forms thereof, unless otherwise indicated or clearly contradicted by the context.

[0215] The term "simulated liquid" of the liquid used to prepare the biological agent means a substitute solution that is similar in physical and chemical properties to the liquid actually used to prepare the biological agent.

[0216] The term "stock solution" of the biological product generally refers to a relatively high concentration solution of the active substance used in the preparation of the drug, which is usually diluted and adjusted as needed during the preparation of the drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0217] Figure 1 The present invention shows a scheme for evaluating shear force during sterile filtration and / or filling.

[0218] Figure 2 The shear force distribution from the center of the pipe to the pipe wall is shown.

[0219] Figure 3 The experimental design of the present invention for the sterilizing filtration process is shown.

[0220] Figure 4 A laboratory device that can be used for the evaluation or optimization method of the present invention is shown, which includes: an experimental pipeline 1, an experimental pump 2, a liquid inlet 3, and a liquid inlet 4.

[0221] Figure 5 A laboratory device that can be used for the evaluation or optimization method of the present invention is shown, wherein liquid can circulate n times in the pipeline, which includes: an experimental pipeline 1, an experimental pump 2, a liquid inlet 3, and a liquid inlet 4. Example

[0222] The following examples are provided to further illustrate the present invention. It should be understood that these examples are provided solely to facilitate a better understanding of the present invention and are not intended to limit the scope of the present invention in any way. Any modifications or equivalent substitutions that do not depart from the spirit of the present invention are intended to be within the scope of the present invention. Unless otherwise specified, the materials, reagents, and apparatus used were obtained from commercial sources or prepared according to methods known in the art.

[0223] Example 1

[0224] In the production of biologics, the liquids used to prepare them need to be sterile-filtered and then filled into containers such as vials through production lines. This example used an aqueous solution of the antibody drug bevacizumab (25 mg / ml, 10 mM histidine-histidine hydrochloride buffer, 8% sucrose, 1 cp) to compare the sample usage and evaluation time of the present method with those of traditional laboratory evaluation methods.

[0225] First, the basic process parameters for sterilization filtration and filling in the biopharmaceutical production scenario were obtained, as shown below:

[0226]

[0227] The average shear force, action time and shear force effects during production are as follows:

[0228]

[0229] 1.1 Traditional laboratory evaluation methods

[0230] According to the proven traditional laboratory evaluation method, the tubing (including length and inner diameter), peristaltic pump, filling peristaltic pump and filling needle that are basically consistent with those in production are selected, and the pump speed is slightly higher than that of production. The specific parameters are as follows:

[0231]

[0232] The minimum sample volume, average shear force, action time, and shear force effect of the above traditional laboratory evaluation method are as follows:

[0233]

[0234] It can be concluded that using traditional laboratory evaluation methods, at least 140 mL of sample is required to complete the experiment, which is costly and time-consuming.

[0235] 1.2 Method of the present invention

[0236] The laboratory parameters of the evaluation method of the present invention are set as follows:

[0237]

[0238]

[0239] For the evaluation method of the present invention, the minimum sample requirement, average shear force, action time and shear force effects are as follows:

[0240] process Minimum sample volume required (mL) Average shear stress (Pa) Time(s) Shear force effect (Pa˙s) Sterile filtration 0.6 5.526 0.543 3.000 Filling 0.4 5.526 1.810 10.000

[0241] As can be seen, the present method only requires 1 mL of sample to fully assess shear forces during sterile filtration and filling, with an action time of just seconds, allowing the experiment to be kept within one minute. Compared to traditional laboratory evaluation methods, the present method significantly reduces sample volume and experiment time, requires fewer consumables, consumes less energy, and requires less manpower.

[0242] Example 2

[0243] Subvisible particles (particle size > 1 μm) are often used to assess protein stability. An increase in the number of subvisible particles generally indicates the possibility of protein aggregation. In this example, an aqueous solution of the antibody drug bevacizumab (same as in Example 1) was used to investigate the reliability and effectiveness of the assessment method of the present invention using subvisible particles.

[0244] According to the verified traditional laboratory evaluation method, process parameters that are basically consistent with the sterilization filtration in the production scenario (the production process parameters are the same as in Example 1) are adopted. At the same time, in order to be able to observe the changes in sub-visible particles, multiple cycles are performed to greatly extend the shear force action time, as shown in the following table.

[0245]

[0246] The laboratory parameter settings of the evaluation method of the present invention, as well as the average shear force and shear force effect are shown in the following table. Similarly, in order to observe the changes of sub-visible particles, multiple cycles were performed to greatly extend the shear force application time.

[0247]

[0248] The antibody solution that has undergone the above production conditions and the antibody solution that has undergone the above laboratory conditions are subjected to subvisible particle detection (see the "Chinese Pharmacopoeia" -0903 Insoluble Particle Test Method, or USP <787> Detection instrument: Tianjin Tianhe GWF-D1. Method: Light obscuration method; 5 injections were performed, the first injection was discarded, and the number of subvisible particles / mL in each of injections 2-5 was measured. The average value was calculated and rounded up, and the data were statistically analyzed. The results are shown in the following table:

[0249] The results of the variance analysis with a significance level of α = 0.05 on the data of sub-visible particles > 2 μm for both groups are as follows:

[0250] Group Number of stitches (after discarding the first stitch) Sum Average number of subvisible particles / mL variance Production Scenario 4 166402.5 41600.63 88803452 Laboratory scene of the present invention 4 195047.5 48761.88 56043.23

[0251] Source of Difference sum of squares F F crit Between groups 1.03E+08 2.308521 5.987378 Within the group 2.67E+08

[0252] The results of the variance analysis with a significance level of α = 0.05 on the data of sub-visible particles > 5 μm for both groups are as follows:

[0253] Group Number of stitches (after discarding the first stitch) Sum Average number of subvisible particles / mL variance Production Scenario 4 87887.5 21971.88 14813731 Laboratory scene of the present invention 4 75515 18878.75 671693.8

[0254] Source of Difference sum of squares F F crit Between groups 19134845 2.471336 5.987378 Within the group 46456273

[0255] From the above variance analysis, it can be seen that F is less than Fcrit, which confirms that the number of subvisible particles obtained in the laboratory scenario of the present invention has no significant difference with that in the production scenario.

[0256] It can be seen that the evaluation method of the present invention almost equivalently simulates the effect of shear force on antibodies in a production scenario, proving that the evaluation method of the present invention is reliable and effective.

[0257] Example 3

[0258] Subvisible particles (particle size > 1 μm) are also used to evaluate the stability of antibody-drug conjugates (ADCs). An increase in the number of subvisible particles is often accompanied by ADC aggregation, fragmentation, isoelectric point changes, and small molecule dropout. In this example, an aqueous solution of the antibody-drug conjugate T-DM1 (trastuzumab-mazine conjugate) (20 mg / ml, 10 mM succinic acid-sodium succinate buffer, 8% sucrose) was used to investigate the reliability and effectiveness of the evaluation method of the present invention using subvisible particles. At the same time, in order to be able to observe changes in subvisible particles, multiple cycles were performed to greatly extend the shear force application time.

[0259] According to the verified traditional laboratory evaluation method, the process parameters are basically the same as those in the production scenario, except that multiple cycles are performed to greatly extend the shear force application time, as shown in the following table

[0260]

[0261]

[0262] The laboratory parameter settings of the evaluation method of the present invention, as well as the average shear force and shear force influence are shown in the following table.

[0263]

[0264] The ADC solution that has undergone the above production conditions and the ADC solution that has undergone the above laboratory conditions are subjected to subvisible particle detection (see the Chinese Pharmacopoeia -0903 Insoluble Particle Test Method, or USP <787> Detection instrument: Tianjin Tianhe GWF-D1. Method: Light obscuration method; 4 injections were made, the first injection was discarded, and the number of subvisible particles / mL for injections 2-4 was measured. The average was calculated and rounded up. The results are shown in the following table:

[0265] The results of the variance analysis with a significance level of α = 0.05 on the data of sub-visible particles > 2 μm for both groups are as follows:

[0266] Group Number of stitches (after discarding the first stitch) Sum Average number of subvisible particles / mL variance Production Scenario 3 6140 2046.667 5877.778 Laboratory scene of the present invention 3 6293.333 2097.778 9181.481

[0267] Source of Difference sum of squares F F crit Between groups 3918.519 0.520413 7.708647 Within the group 30118.52

[0268] The results of the variance analysis with a significance level of α = 0.05 on the data of sub-visible particles > 5 μm for both groups are as follows:

[0269] Group Number of stitches (after discarding the first stitch) Sum Average number of subvisible particles / mL variance Production Scenario 3 2716.667 905.5556 414.8148 Laboratory scene of the present invention 3 3130 1043.333 7811.111

[0270] Source of Difference sum of squares F F crit Between groups 28474.07 6.923008 7.708647 Within the group 16451.85

[0271] From the above variance analysis, it can be seen that F is less than Fcrit, which confirms that the number of subvisible particles obtained in the laboratory scenario of the present invention has no significant difference with that in the production scenario.

[0272] As can be seen, the method of the present invention is also applicable to evaluating the stability of ADC under shear stress. This method almost equivalently simulates the effect of shear stress on ADC during production, demonstrating that the evaluation method of the present invention is reliable and effective.

[0273] All publications, patent applications, patents and other references mentioned herein are incorporated herein by reference in their entirety. Any or all features discussed above and throughout this application may be combined in various embodiments of the present invention. In addition, the materials, methods and examples described herein are merely illustrative and are not intended to be limiting.

Claims

1. A method for evaluating the effect of shear stress on the stability of a biological agent during production, comprising: (a) Obtaining geometric parameters of the production passage and operating parameters of the production pump in the production scene; (b) determining geometric parameters of the experimental path and operating parameters of the experimental pump in the laboratory scenario based on the production scenario parameters obtained in step (a); (c) determining the required sample usage and evaluation time based on the production scenario parameters obtained in step (a) and the laboratory scenario parameters determined in step (b); (d) running the evaluation time determined in step (c) in a laboratory scenario having the laboratory scenario parameters determined in step (b) using the drug liquid or a simulated liquid thereof for preparing the biological agent in the sample usage amount determined in step (c); and (e) detecting changes in the stability of the liquid obtained in step (d) to evaluate the effect of shear force on the stability of the biological preparation.

2. A method for optimizing a biological preparation production process, comprising: (a) Setting the geometric parameters of the production path and the operating parameters of the production pump in the production scenario as required; (b) determining the geometric parameters of the experimental path and the operating parameters of the experimental pump in the laboratory scenario based on the production scenario parameters set in step (a); (c) determining the required sample usage and evaluation time based on the production scenario parameters set in step (a) and the laboratory scenario parameters determined in step (b); (d) running the evaluation time determined in step (c) in a laboratory scenario having the laboratory scenario parameters determined in step (b) using the drug liquid or a simulated liquid thereof for preparing the biological agent in the sample usage amount determined in step (c); (e) detecting a change in the stability of the liquid obtained in step (d) to evaluate the effect of shear force on the stability of the biological preparation; and (f) When the detection of step (e) shows that the effect of shear force on the stability of the biological agent is acceptable, the geometric parameters of the production path and the operating parameters of the production pump in the corresponding production scenario are determined as the production process of the biological agent; when the detection of step (e) shows that the effect of shear force on the stability of the biological agent is unacceptable, the geometric parameters of the production path and the operating parameters of the production pump in the production scenario set in step (a) are adjusted, and steps (b) to (e) are repeated until the detection of step (e) shows that the effect of shear force on the stability of the biological agent is acceptable or desired.

3. The method according to claim 1 or 2, wherein The production path includes a production line for conveying liquid (e.g., liquid for preparing the biological agent) and an internal channel of an optional filling needle for filling the liquid; optionally, the production line is substantially composed of a silicone tube; and / or The experimental passage is an experimental pipeline for transporting or circulating liquid (eg, liquid for preparing the biological agent or a simulated liquid thereof); optionally, the experimental passage is substantially composed of a silicone tube.

4. A method according to any one of the preceding claims, wherein: The geometric parameters of the production passage include the length L of the production passages with different radii. i and radius R i The working parameters of the production pump include the production pump speed PR of the production channels with different radii. i and the ratio of production flow to production pump speed k i , where i is any integer from 1 to m, and m is the number of radius values ​​in the production path; and The geometric parameters of the experimental path include the length L of the experimental path lab and radius R lab The working parameters of the experimental pump include the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab .

5. A method according to any one of the preceding claims, wherein The geometric parameters of the experimental passage and the operating parameters of the experimental pump are determined based on the following conditions: (i) Experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than the production pump speed PR of each production channel with different radius i and the ratio of production flow to production pump speed k i The product of divided by the radius R of the production channel i The quotient of the cube of (PR i ·k i ) / R i 3 The maximum value in (ii) Optionally, the length L of the experimental path lab With radius R lab The ratio L lab / R lab is the length L of the production channels with different radii i With radius R i The ratio L i / R i The sum of about 0.8 to about 10 times of; wherein i is any integer from 1 to m, and m is the number of radius values ​​in the production path; and Among them, the length of the experimental path L lab It refers to the distance that a liquid travels in a device in a laboratory setting.

6. The method according to claim 5, wherein: For (i), the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 is the production pump speed PR of production channels with different radii i and the ratio of production flow to production pump speed k i The product of divided by the radius R of the production channel i The quotient of the cube of (PR i ·k i ) / R i 3 1 to about 100 times, e.g., about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times, the maximum value in ; and / or For (ii), the length of the experimental path L lab With radius R lab The ratio L lab / R lab is the length L of the production channels with different radii i With radius R i The ratio L i / R i The sum of The amount of the active ingredient is about 0.8 to about 5 times, for example, about 0.8 to about 2 times, about 1 to about 1.5 times, or about 1 to about 1.2 times.

7. A method according to any one of the preceding claims, wherein In step (c) The lengths L of the production passages of different radii are based on i and radius R i , the radius R of the experimental path lab , the experimental pump speed PR lab and the ratio k of the experimental flow rate to the experimental pump speed lab To determine the evaluation time t lab Preferably, the evaluation time t lab yes about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 times; and / or Based on the length L of the experimental path lab and radius R lab and determining the sample usage of the biological agent based on the number of cycles n of the sample in the device of the laboratory scene, wherein n is any integer or decimal greater than or equal to 1; optionally, the minimum sample usage of the biological agent is (πR lab 2 L lab ) / n, where n is any integer or decimal greater than or equal to 1.

8. A method according to any one of the preceding claims, wherein The evaluation method is used to evaluate the effect of shear force on the stability of a biological agent during a filtration sterilization operation of the biological agent, and the optimization method is used to optimize the production process of the filtration sterilization operation of the biological agent; wherein the production passage is uniform in thickness, and the experimental passage is uniform in thickness; optionally, the production passage and the experimental passage are substantially composed of silicone tubing; The geometric parameters of the production passage include the length L of the production passage. filt and radius R filt The working parameters of the production pump include the production pump speed PR filt and the ratio of production flow to production pump speed k filt The geometric parameters of the experimental path include the length L of the experimental path lab and radius R lab The working parameters of the experimental pump include the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab .

9. The method according to claim 8, wherein The geometric parameters of the experimental passage and the operating parameters of the experimental pump are determined based on the following conditions: (i) Experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than production pump speed PR filt and the ratio of production flow to production pump speed k filt The product of divided by the radius R of the production channel filt The quotient of the cube of (PR filt ·k filt ) / R filt 3 ,Right now and (ii) Optionally, the length L of the experimental path lab With radius R lab The ratio L lab / R lab is the length of the production channel L filt With radius R filt The ratio L filt / R filt about 0.8 to about 10 times of; Among them, the length of the experimental path L lab It refers to the distance that a liquid travels in a device in a laboratory setting.

10. The method according to claim 9, wherein: For (i), the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 is the production pump speed PR filt and the ratio of production flow to production pump speed k filt The product of divided by the radius R of the production channel filt The quotient of the cube of (PR filt ·k filt ) / R filt 3 about 1 to about 100 times, e.g., about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times; and / or For (ii), the length of the experimental path L lab With radius R lab The ratio L lab / R lab is the length of the production channel L filt With radius R filt The ratio L filt / R filt The amount of the active ingredient is about 0.8 to about 5 times, for example, about 0.8 to about 2 times, about 1 to about 1.5 times, or about 1 to about 1.2 times.

11. The method according to any one of claims 8 to 10, wherein: In step (c) Based on the length L of the production path filt and radius R filt , the radius R of the experimental path lab , the experimental pump speed PR lab and the ratio k of the experimental flow rate to the experimental pump speed lab To determine the evaluation time t lab Preferably, the evaluation time t lab yes about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 times; and / or Based on the length L of the experimental path lab and radius R lab and determining the sample usage of the biological agent based on the number of cycles n of the sample in the device of the laboratory scene, wherein n is any integer or decimal greater than or equal to 1; optionally, the minimum sample usage of the biological agent is (πR lab 2 L lab ) / n, where n is any integer or decimal greater than or equal to 1.

12. The method according to claim 1, wherein The evaluation method is used to evaluate the effect of shear force on the stability of a biologic during a filling operation of the biologic, and the optimization method is used to optimize the production process of the filling operation of the biologic; wherein the production pipeline for conveying liquid is uniform in thickness, and the experimental passage is uniform in thickness; and The geometric parameters of the production path include the length L of the production pipeline. fill and radius R fill And the length of the filling needle L needle and radius R needle The working parameters of the production pump include the production pump speed PR fill and the ratio of production flow to production pump speed k fill The geometric parameters of the experimental path include the length L of the experimental path lab and radius R lab The working parameters of the experimental pump include the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab .

13. The method according to claim 12, wherein: The geometric parameters of the experimental passage and the operating parameters of the experimental pump are determined based on the following conditions: (i) Experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill The product of divided by the radius R of the production pipeline fill The quotient of the cube of (PR fill ·k fill ) / R fill 3 and filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·k fill ) / R fill 3 The maximum value in and (ii) Optionally, the length L of the experimental path lab With radius R lab The ratio L lab / R lab is the length of the production pipeline L fill and radius R fill The ratio of the filling needle length L needle and radius R needle The sum of the ratios L fill / R fill +L needle / R needle about 0.8 to about 10 times of; Among them, the length of the experimental path L lab It refers to the distance that a liquid travels in a device in a laboratory setting.

14. The method according to claim 13, wherein: For (i), the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Is the filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill The product of divided by the radius R of the production pipeline fill The quotient of the cube of (PR fill ·k fill ) / R fill 3 Filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·k fill ) / R needle 3 1 to about 100 times, e.g., about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times, the maximum value in ; and / or For (ii), the length L of the experimental path lab With radius R lab The ratio L lab / R lab is the length L of the production pipeline fill and radius R fill The ratio of the filling needle length L needle and radius R needle The sum of the ratios L fill / R fill +L needle / R needle The amount of the active ingredient is about 0.8 to about 5 times, for example, about 0.8 to about 2 times, about 1 to about 1.5 times, or about 1 to about 1.2 times.

15. The method according to claim 13 or 14, wherein condition (i) is: the experimental pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Not less than filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·k fill ) / R fill 3 ,Right now Optionally, test pump speed PR lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 Is the filling pump speed PR fill and the ratio of filling flow to filling pump speed k fill divided by the filling needle radius R needle The quotient of the cube of (PR fill ·k fill ) / R needle 3 The amount of the active ingredient is about 1 to about 100 times, for example, about 1 to about 75 times, about 1 to about 50 times, or about 1 to about 25 times, or about 1 to about 10 times.

16. The method according to any one of claims 12 to 15, wherein: Based on the length L of the production pipeline fill and radius R fill And the length of the filling needle L needle and radius R needle , the radius R of the experimental path lab , the experimental pump speed PR lab and the ratio k of the experimental flow rate to the experimental pump speed lab To determine the evaluation time t lab Preferably, the evaluation time t lab yes about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 times; and / or Based on the length L of the experimental path lab and radius R lab and determining the sample usage of the biological agent based on the number of cycles n of the sample in the device of the laboratory scene, wherein n is any integer or decimal greater than or equal to 1; optionally, the minimum sample usage of the biological agent is (πR lab 2 L lab ) / n, where n is any integer or decimal greater than or equal to 1, representing the number of cycles of the sample in the experimental pathway.

17. The method according to any one of claims 1 to 16, wherein the laboratory background comprises more than one pump in parallel. Experimental pump speed PR in condition (i) lab and the ratio of the experimental flow rate to the experimental pump speed k lab The product of divided by the radius R of the experimental path lab The quotient of the cube of (PR lab ·k lab ) / R lab 3 represents the sum of the quotients of each pump, that is, and The evaluation time t lab yes about 0.8 to about 20 times, preferably about 0.8 to about 10 times, more preferably about 0.8 to about 5 times, still more preferably about 0.8 to about 1.5 times, for example about 1 time, in, PR lab,i is the pump speed of the i-th pump in the laboratory, k lab,i is the pump flow / pump speed of the ith pump in the laboratory, num_pump is the number of pumps in the laboratory, L i and R i are the length and radius of production channels with different radii, and m is the number of radius values ​​in the production channel.

18. The method according to any one of the preceding claims, wherein the biologic is susceptible to aggregation or denaturation under the action of shear force; optionally, the biopharmaceutical molecules of the biologic are susceptible to aggregation or denaturation under the action of shear force; optionally, the biopharmaceutical molecules of the biologic are selected from one or more of proteins, polypeptides, nucleic acids, or any derivatives thereof (e.g., fusions or conjugates), for example, one or more of antibodies (e.g., monoclonal antibodies or polyclonal antibodies) or antigen-binding fragments thereof, fusion proteins, bioconjugate drugs (XDCs) such as antibody-drug conjugates (ADCs), polypeptides, and small nucleic acid drugs.

19. The method according to any one of the preceding claims, wherein the biologic is an injectable biologic; optionally, the biologic is in the form of a liquid formulation or a spray-dried powder or a freeze-dried powder.

20. The method according to any one of the preceding claims, wherein the pharmaceutical liquid used to prepare the biologic is a solution comprising the biopharmaceutical molecules of the biologic, such as an aqueous solution, which optionally contains one or more excipients; or, the pharmaceutical liquid used to prepare the biologic is a pharmaceutical stock solution of the biologic.

21. The method according to any of the preceding claims, wherein in step (e), the assessment is achieved by detecting changes in physical stability and / or chemical stability of the biopharmaceutical molecule; optionally, the instability is manifested as one or more of aggregation, denaturation, adsorption, precipitation, hydrolysis, degradation and / or oxidation; optionally, the stability test items include one or more of the following: appearance, including clarity or turbidity, color, visible particles; sub-visible particles; pH value; biopharmaceutical molecule concentration; biopharmaceutical molecule molecular weight; viscosity; biological activity; purity; degradation product content; charge variants; for antibody drug conjugates, average Dar value and / or free toxins.

22. A laboratory device for use in the method according to any one of the preceding claims, comprising a laboratory line 1 and a laboratory pump 2 in fluid communication with each other, and having a liquid inlet 3 and a liquid outlet 4 on the laboratory line, wherein the laboratory line 1 has geometrical parameters determined according to the method according to any one of claims 1 to 17, and the laboratory pump 2 is configured to have operating parameters determined according to the method according to any one of claims 1 to 17; Optionally, the experimental tubing is made of the same material as the production tubing, for example, substantially consisting of silicone tubing, and the experimental pump is a peristaltic pump, for example, a peristaltic hose pump; Optionally, the laboratory device further comprises one or more elements selected from the following: a liquid supply element, a liquid receiving element, a metering element, a timing element, a switching element, a parameter monitoring and / or control element (e.g., a pump speed monitoring and / or control element, a flow monitoring and / or control element, a shear force monitoring and / or control element, a viscosity monitoring and / or control element, a temperature monitoring and / or control element), a cycle number control and / or recording element, and a stability detection element.

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