CN116337697A - A method and system for detecting the empty packet ratio of an adeno-associated virus vector - Google Patents

Abstract

The invention provides a method and a system for detecting the blank ratio of an adeno-associated virus vector, which are characterized in that the sedimentation coefficient value of the adeno-associated virus vector and the sedimentation distribution coefficient value corresponding to the sedimentation coefficient value are measured through an analytical ultracentrifuge, the sedimentation coefficient value and the sedimentation distribution coefficient value are normalized, and the blank ratio is obtained through the ratio of each sedimentation distribution coefficient value to the sum of the sedimentation distribution coefficient values. According to the method and the system for detecting the blank ratio of the adeno-associated virus vector, disclosed by the embodiment of the invention, the sedimentation coefficient value and the sedimentation distribution coefficient value of the adeno-associated virus vector are measured through the analytical ultracentrifuge, the sedimentation coefficient value and the sedimentation distribution coefficient value are normalized, the blank ratio is obtained through the ratio of each sedimentation distribution coefficient value to the sum of the sedimentation distribution coefficient values, and the detection effect on the blank ratio of the adeno-associated virus vector is greatly improved.

Description

A method and system for detecting the empty packet ratio of an adeno-associated virus vector

technical field

The invention relates to the field of gene detection, in particular to a method for detecting the empty packet ratio of an adeno-associated virus vector; in addition, the invention also relates to a system for detecting the empty packet ratio of the adeno-associated virus vector.

Background technique

In the field of gene therapy, adeno-associated virus vectors are considered to be one of the most promising gene delivery vectors. In the production process of adeno-associated virus vectors, product quality homogeneity, vector purity and batch consistency are important aspects of quality control. In addition to ensuring the titer, yield and biological activity of adeno-associated virus vectors, the most important thing is to improve Purity of the adeno-associated virus vector. Among them, the most important task is to remove defective adeno-associated virus vectors, that is, to remove empty capsid virus particles or virus vector particles containing DNA other than full-length genes. There is a lack of standard methods to ensure the quality and effectiveness of adeno-associated virus vector products at the physical, chemical and biological levels, and there is also a lack of more effective detection methods to detect the empty packet ratio of adeno-associated virus vectors in the prior art.

Contents of the invention

In order to solve the problems existing in the prior art, at least one embodiment of the present invention provides a method for detecting the empty packet ratio of the adeno-associated virus vector, which greatly improves the detection effect of the empty packet ratio of the adeno-associated virus vector . To this end, at least one embodiment of the present invention also provides a system for detecting the empty packet ratio of an adeno-associated virus vector.

In the first aspect, the embodiment of the present invention proposes a method for detecting the empty packet ratio of an adeno-associated virus vector, the method comprising the following steps:

The sedimentation coefficient value and the corresponding sedimentation distribution coefficient value of the adeno-associated virus vector are determined by an analytical ultracentrifuge;

Normalize the sedimentation coefficient value and the sedimentation distribution coefficient value;

The empty bag ratio is obtained by the ratio of each settlement distribution coefficient value to the sum of the settlement distribution coefficient values.

In some embodiments, the present invention provides a method for detecting the empty packet ratio of an adeno-associated virus vector. Obtaining the sedimentation coefficient, molar mass and diffusion coefficient value through the sedimentation curve measured by the analytical ultracentrifuge includes the following steps:

Use the c(s) analysis model of SEDFIT software, all components in the adeno-associated virus carrier solution have the same coefficient of friction ratio in this model, calculate the following formula:

f ₀ =6πηr ₀ ,

Wherein, f ₀ is the friction coefficient that macromolecule collapses into compact bead, η is solvent viscosity, r ₀ is hydrodynamic radius;

Transform the following formula:

Wherein, M is the molar mass, N _A is the number of moles, and _ρ is the particle density of the solution;

get:

where ν is the specific volume of the solute;

According to the Svedberg equation:

Where s is the value of the sedimentation coefficient;

Get the molar mass:

Substitute the sedimentation coefficient and the corresponding molar mass into the Svedberg equation to obtain the value of the diffusion coefficient:

Among them, D is the diffusion coefficient value, R is the gas constant, T is the absolute temperature, and ρ is the solvent density.

In some embodiments, the method for detecting the empty packet ratio of an adeno-associated virus vector provided by the present invention, obtaining the sedimentation coefficient value and the sedimentation distribution coefficient value includes the following steps:

The data measured by the analytical ultracentrifuge are processed by SEDFIT software;

The value of the sedimentation coefficient and the value of the sedimentation distribution coefficient are obtained by integrating the processed data through the data of each peak displayed by the GUSSI software.

In some embodiments, the present invention provides a method for detecting the empty envelope ratio of an adeno-associated virus vector, wherein the reference substance of the adeno-associated virus vector is the preparation buffer stored in the adeno-associated virus vector virus liquid.

In some embodiments, the present invention provides a method for detecting the empty envelope ratio of an adeno-associated virus vector, and the preparation buffer includes water, saline solution and cell culture medium.

In some embodiments, the present invention provides a method for detecting the empty packet ratio of an adeno-associated virus vector, and the detection temperature of the analytical ultracentrifuge is 15°C-25°C.

In some embodiments, the present invention provides a method for detecting the empty packet ratio of an adeno-associated virus vector, wherein the absorbance of the adeno-associated virus vector is 0.3-1.5.

In the second aspect, the embodiment of the present invention also provides an adeno-associated virus vector empty packet ratio detection system, including:

The determination module measures the sedimentation coefficient value of the adeno-associated virus vector and the corresponding sedimentation distribution coefficient value by an analytical ultracentrifuge;

A normalization processing module is used for normalizing the sedimentation coefficient value and the sedimentation distribution coefficient value;

The empty bag ratio calculation module is used to calculate the empty bag ratio through the ratio of each settlement distribution coefficient value to the sum of the settlement distribution coefficient values.

In some embodiments, an adeno-associated virus vector empty packet ratio detection system provided by the present invention further includes:

The parameter setting module is used for setting the parameters of the analytical ultracentrifuge.

In the third aspect, the embodiment of the present invention also provides an adeno-associated virus vector empty packet ratio detection device, including at least one processor; a memory coupled to the at least one processor, the memory stores executable instructions, and the executable instructions are executed At least one processor implements the steps of any one method of the above first aspect when executed.

In a fourth aspect, an embodiment of the present invention further provides a chip configured to execute the steps of the method in the first aspect above. Specifically, the chip includes: a processor, configured to invoke and run a computer program from the memory, so that the device installed with the chip is used to execute the steps of the method in the first aspect above.

In the fifth aspect, the embodiment of the present invention also provides a computer-readable storage medium, on which a computer program is stored, and when the computer program is executed by a processor, the steps of any one of the methods in the first aspect above are implemented.

In a sixth aspect, an embodiment of the present invention further provides a computer program product, including computer program instructions, where the computer program instructions cause a computer to execute the steps of the method in the first aspect above.

It can be seen that in the method and system for detecting the empty packet ratio of an adeno-associated virus vector according to the embodiment of the present invention, the sedimentation coefficient value and the sedimentation distribution coefficient value of the adeno-associated virus vector are measured by an analytical ultracentrifuge, and then normalized , the empty packet ratio is obtained by the ratio of each sedimentation distribution coefficient value to the sum of the sedimentation distribution coefficient values, which greatly improves the detection effect of the empty packet ratio of the adeno-associated virus vector.

Description of drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the accompanying drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only some of the present invention. Embodiments, for those of ordinary skill in the art, other drawings can also be obtained according to these drawings without paying creative labor.

Fig. 1 is a flow chart showing a method for detecting the empty packet ratio of an adeno-associated virus vector in an embodiment of the present invention;

Figure 2 shows a schematic diagram of the sedimentation curve of the adeno-associated virus vector stored at -80°C in the embodiment of the present invention;

Figure 3 shows a schematic diagram of the sedimentation curve of the adeno-associated virus vector in the embodiment of the present invention after being frozen and thawed three times;

Figure 4 shows a schematic diagram of the sedimentation curve of the adeno-associated virus vector in the embodiment of the present invention placed at room temperature for 72 hours;

Fig. 5 is a schematic diagram showing the empty packet ratio analysis of the adeno-associated virus vector in the embodiment of the present invention when the preservation detection wavelength λ = 220nm under four different conditions;

Figure 6 is a schematic diagram of the sedimentation curve of the adeno-associated virus vector in the first sedimentation cycle at 15000rpm in the embodiment of the present invention;

Figure 7 is a schematic diagram of the sedimentation curve of the adeno-associated virus vector at 20000rpm in the embodiment of the present invention;

Fig. 8 shows a schematic diagram of the sedimentation curve when the adeno-associated virus vector is diluted to the same multiple in the concentration of 15000 rpm and OD=0.3-1.5 in the embodiment of the present invention;

Figure 9 is a schematic diagram showing the empty packet ratio analysis of the adeno-associated virus vector at 15000rpm and 20000rpm in the embodiment of the present invention;

Fig. 10 is a schematic diagram of an empty packet ratio detection system for an adeno-associated virus vector in an embodiment of the present invention.

specific implementation plan

In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments It is a part of embodiments of the present invention, but not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

It should be noted that in this article, relative terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply these No such actual relationship or order exists between entities or operations. As used herein, the terms "comprises," "comprises," or any other variation thereof are intended to encompass a non-exclusive inclusion such that a process, method, article, or apparatus that includes a set of elements includes not only those elements, but also includes the elements not expressly included. other elements listed, or also include elements inherent in such a process, method, article, or device. Without further limitations, an element defined by the phrase "comprising a ..." does not preclude the presence of additional identical elements in the process, method, article, or apparatus comprising that element.

Adeno-associated virus is a kind of single-stranded linear DNA defective virus. Its genomic DNA is less than 5kb, without envelope, and its appearance is naked icosahedral particles. It can be understood that the adeno-associated virus has no special limitation on its outer envelope protein. Therefore, the adeno-associated viral vector according to the present invention may be of any serotype.

Adeno-associated virus is a liquid of virus stored in formulation buffer. Formulation buffer refers to a substance that, in this case, keeps the virus alive. Formulation buffers used in the context of the present invention are water, saline solutions, cell culture media, and the like.

Empty packet ratio can be understood as the ratio of empty capsid virus particles, virus particles packaged with part of the genome, complete adeno-associated virus vectors packaged with the required full-length target gene, and genome overloaded virus particles to the total number of virus particles.

The inventors of this solution found that in the prior art, there is a lack of more effective detection means to detect the empty packet ratio of the adeno-associated virus vector. Embodiments of the present invention provide the following solutions:

Sedimentation velocity experiments were performed using an Optima AUC analytical ultracentrifuge using a dual sector epoxy (epon) centerpiece with a 12 mm path length. Place the sample cell in an An-60Ti four-hole rotor or An-50 Ti eight-hole rotor. Typically, a sedimentation velocity curve scan is performed after equilibrating for a period of time without imparting rotational speed to the analytical ultracentrifuge. Subsequently, a rotor speed of 15,000-20,000 rpm was used for the sedimentation velocity experiment of the adeno-associated virus vector. Fill both sectors of the centerpiece with approximately 380-400 μl of sample and approximately 400-440 μl of formulation buffer as reference. All sedimentation experiments were carried out at a temperature of T = 15-25°C. Sedimentation curve scans were recorded at λ = 220-230 nm using an absorbance detection system.

As shown in Figure 1, in the first aspect, the embodiment of the present invention provides a method for detecting the empty packet ratio of an adeno-associated virus vector, the method includes the following steps:

The relevant parameters of the analytical ultracentrifuge were selected according to the adeno-associated virus vector sample, and the relevant parameters included rotor speed, distance between the sample and the center of the rotor, temperature, buffer, centrifugation time, detection time interval, sector and optical window characteristics.

The sedimentation coefficient value and the corresponding sedimentation distribution coefficient value of each component in the adeno-associated virus vector were determined by an analytical ultracentrifuge.

Sedimentation coefficient values are derived from measurements of the rate of movement of the sedimentation boundary in analytical ultracentrifuge experiments. The value of the sedimentation coefficient depends on the shape of the particle, where in particles whose shape deviates from a sphere, the friction ratio f/f0 is greater than 1 (friction coefficient f of the molecule in question/friction coefficient f0 of a smooth sphere with the same anhydrous volume and mass) sedimentation The coefficient further depends on the molecular weight/molar mass of the particles, with larger particles settling faster. Sedimentation coefficient values can be measured by looking at populations of subsiding species at suitable time intervals; as above, during subsidence a boundary is created that progresses towards higher radial values over time.

The sedimentation distribution coefficient represents the sedimentation interface of the corresponding component. Because the sample particles are very small, their sedimentation motion cannot be seen directly, so the interface moving speed of the sample particles during centrifugation is regarded as the average sedimentation speed of the sample particles. Interfacial sedimentation maps are typically recorded using the Schlieren effect and absorption optics. The sample interface in the sedimentation diagram generally shows a symmetrical peak, and the highest point of the peak represents the interface position. Points are not enough as the interface position, and the second-order distance method should be used to calculate the interface position.

Initial scans and changes in sedimentation curves during the sedimentation velocity experiments monitored at λ = 220 nm for adeno-associated viral vectors. Experiments were performed at 15,000 rpm. As shown in Figure 2, the adeno-associated virus vector was stored at -80°C. As shown in Fig. 3, the adeno-associated virus vector was frozen and thawed three times, and diluted the same times in the concentration of OD=0.3-1.5. The experimental curves were analyzed using the finite element method, combined with maximum entropy regularization. The model assumes that all components in a solution have the same ratio of coefficients of friction (f/f ₀ ). Obtaining the sedimentation coefficient, molar mass and diffusion coefficient values from the sedimentation curve determined by the analytical ultracentrifuge includes the following steps:

Use the c(s) analysis model of SEDFIT software, all components in the adeno-associated virus carrier solution have the same coefficient of friction ratio in this model, calculate the following formula:

f ₀ =6πηr ₀ ,

Wherein, f ₀ is the friction coefficient that macromolecule collapses into compact bead, η is solvent viscosity, r ₀ is hydrodynamic radius;

Transform the following formula:

Wherein, M is the molar mass, N _A is the number of moles, and _ρ is the particle density of the solution;

get:

where ν is the specific volume of the solute;

According to the Svedberg equation:

Where s is the value of the sedimentation coefficient;

Get the molar mass:

Substitute the sedimentation coefficient and the corresponding molar mass into the Svedberg equation to obtain the value of the diffusion coefficient:

Among them, D is the diffusion coefficient value, R is the gas constant, T is the absolute temperature, and ρ is the solvent density.

Sedimentation coefficient values have a distinct differential distribution, representing groups of subsiding species. As can be seen from the residual plot, only a few percent fluctuations are shown.

The set of experiments just described was performed with freshly received adeno-associated virus vectors after preparation and purification (almost no storage), and at room temperature for 72 hours, as shown in Figure 4 . When the detection wavelength λ=220nm, the dilution factor is the same as that of the sample preserved under the -80°C condition. There are four kinds of analysis of the empty envelope ratio of the adeno-associated virus vector preserved under different conditions, respectively after preparation and purification ( Almost no storage) freshly received adeno-associated virus vectors, adeno-associated virus vectors that have been tested once and tested again, adeno-associated virus vectors that have been frozen and thawed three times, and adeno-associated virus vectors that have been placed at room temperature for 72 hours, as shown in Figure 5 Show. Obtaining the sedimentation coefficient value and the sedimentation distribution coefficient value includes the following steps: the data measured by the analytical ultracentrifuge is processed by SEDFIT software; the processed data is obtained by integrating the data of each peak displayed by the GUSSI software to obtain the sedimentation coefficient value and the value of the sedimentation distribution coefficient.

The sedimentation coefficient value and the sedimentation distribution coefficient value are normalized. Map the sedimentation distribution coefficient value of each sedimentation coefficient value of the adeno-associated virus vector to the range of (0,1) for processing, and obtain the empty packet ratio by the ratio of each sedimentation distribution coefficient value to the sum of the sedimentation distribution coefficient values, that is, the adeno-associated virus The ratio of the sedimentation distribution coefficient value of the carrier at each sedimentation coefficient value to the sum of the sedimentation distribution coefficient values of the sedimentation coefficient value between 20-180.

An advantage of the analytical ultracentrifuge is that it preserves the mass balance in the closed sector cell solvent. As shown in Figure 6, the experiment in the first settling cycle at 15,000 rpm, after which the sample cell was removed from the centrifuge rotor and shaken to achieve resuspension of the settled material. Afterwards, another settling velocity experiment was performed under exactly the same conditions, followed by a corresponding settling analysis. The results obtained are very similar, demonstrating the reproducibility of the results obtained using the solutions at specific storage times and that the experiments are repeatable.

As shown in Figure 7, the adeno-associated virus vector was at 20,000rpm; as shown in Figure 8, at 15000rpm, and diluted the same multiple in the concentration of OD=0.3-1.5, the initial sedimentation velocity during the experiment monitored at λ=220nm Changes in sweep and sedimentation curves. The results of the empty packet ratio analysis are shown in Fig. 9, which also indicated that the sedimentation at 15000-20000rpm did not significantly affect the particle integrity and the estimated signal intensity of the AAV vector.

As shown in FIG. 10 , in the second aspect, the embodiment of the present invention also provides an adeno-associated virus vector empty packet ratio detection system, including a parameter setting module, a measurement module, a normalization processing module and an empty packet ratio calculation module. The parameter setting module is used to set the parameters of the analytical ultracentrifuge, the determination module is used to determine the sedimentation coefficient value of the adeno-associated virus vector and the corresponding sedimentation distribution coefficient value through the analytical ultracentrifuge, and the normalization processing module is used In order to normalize the sedimentation coefficient value and the sedimentation distribution coefficient value, the empty envelope ratio calculation module is used to calculate the empty envelope ratio through the ratio of each sedimentation distribution coefficient value to the sum of the sedimentation distribution coefficient values.

In the third aspect, the embodiment of the present invention also provides an adeno-associated virus vector empty packet ratio detection device, including:

At least one processor; a memory coupled to the at least one processor, the memory storing executable instructions, wherein the executable instructions, when executed by the at least one processor, cause the implementation of the method steps of the first aspect of the present invention.

In the device for detecting the empty packet ratio of an adeno-associated virus vector provided by an embodiment of the present invention, the processor and the memory can be set separately or integrated together.

For example, the memory may include random access memory, flash memory, read-only memory, programmable read-only memory, non-volatile memory or registers, and the like. The processor may be a central processing unit (Central Processing Unit, CPU) or the like. Or the image processor (Graphic Processing Unit, GPU) memory can store executable instructions. A processor may execute executable instructions stored in memory to implement the various processes described herein.

It can be understood that the memory in this embodiment may be a volatile memory or a nonvolatile memory, or may include both volatile and nonvolatile memories. Among them, the non-volatile memory can be ROM (Read-Only Memory, read-only memory), PROM (ProgrammableROM, programmable read-only memory), EPROM (ErasablePROM, erasable programmable read-only memory), EEPROM (Electrically Erasable Programmable Read-Only Memory) or Flash. The volatile memory may be RAM (Random Access Memory, random access memory), which is used as an external cache. By way of illustration and not limitation, many forms of RAM are available such as SRAM (Static RAM, Static Random Access Memory), DRAM (Dynamic RAM, Dynamic Random Access Memory), SDRAM (Synchronous DRAM, Synchronous Dynamic Random Access Memory), DDRSDRAM (DoubleDataRate SDRAM, double data rate synchronous dynamic random access memory), ESDRAM (Enhanced SDRAM, enhanced synchronous dynamic random access memory), SLDRAM (SynchlinkDRAM, synchronous connection dynamic random access memory) and DRRAM (DirectRambusRAM, direct memory bus random access memory). The memories described herein are intended to include, but are not limited to, these and any other suitable types of memories.

In some embodiments, the memory stores the following elements, upgrade packages, executable units or data structures, or a subset thereof, or an extended set thereof: an operating system and an application program.

Among them, the operating system includes various system programs, such as framework layer, core library layer, driver layer, etc., which are used to realize various basic services and process hardware-based tasks. The application program includes various application programs and is used to implement various application services. Programs for implementing the methods of the embodiments of the present invention may be contained in application programs.

In the embodiment of the present invention, the processor invokes the program or instruction stored in the memory, specifically, the program or instruction stored in the application program, and the processor is configured to execute the method steps provided in the first aspect.

In a fourth aspect, an embodiment of the present invention further provides a chip configured to execute the method in the first aspect above. Specifically, the chip includes: a processor, configured to call and run a computer program from the memory, so that the device installed with the chip is used to execute the method in the first aspect above.

In the fifth aspect, the embodiment of the present invention also provides a computer-readable storage medium, on which a computer program is stored, and when the computer program is executed by a processor, the steps of the method in the first aspect of the present invention are implemented.

For example, a machine-readable storage medium may include, but is not limited to, various known and unknown types of nonvolatile memory.

In a sixth aspect, an embodiment of the present invention further provides a computer program product, including computer program instructions, where the computer program instructions cause a computer to execute the method in the first aspect above.

To sum up, in the embodiment of the present invention, an adeno-associated virus vector empty packet ratio detection method and system uses an analytical ultracentrifuge to measure the sedimentation coefficient value and sedimentation distribution coefficient value of the adeno-associated virus vector, and normalizes them. In the normalization process, the empty packet ratio is obtained by the ratio of each sedimentation distribution coefficient value to the sum of the sedimentation distribution coefficient values, which greatly improves the detection effect of the empty packet ratio of the adeno-associated virus vector.

Those skilled in the art can understand that the units and algorithm steps of the examples described in conjunction with the embodiments disclosed herein can be implemented by electronic hardware, or a combination of software and electronic hardware. Whether these functions are implemented in hardware or software depends on the specific application and design constraints of the technical solution. Those skilled in the art may implement the described functions in different ways for each specific application, but such implementation should not be considered as exceeding the scope of the present application.

In the embodiments of the present application, the disclosed systems, devices and methods may be implemented in other ways. For example, the division of units is only a logical function division, and there may be other division methods in actual implementation. For example, multiple units or components may be combined or integrated into another system. In addition, the coupling between the respective units may be direct coupling or indirect coupling. In addition, each functional unit in the embodiment of the present application may be integrated into one processing unit, or may exist independently, etc. physically.

It should be understood that in various embodiments of the present application, the sequence numbers of the processes do not mean the order of execution, and the execution order of the processes should be determined by their functions and internal logic, rather than by the embodiments of the present application. The implementation process constitutes any limitation.

If the functions are realized in the form of software function units and sold or used as an independent product, they can be stored in a machine-readable storage medium. Therefore, the technical solution of the present application can be embodied in the form of a software product, and the software product can be stored in a machine-readable storage medium, which can include several instructions to make the electronic device execute the technical solution described in the embodiment of the present application. all or part of the process. The above-mentioned storage medium may include ROM, RAM, removable disk, hard disk, magnetic disk or optical disk, and other various media that can store program codes.

The above content is only a specific implementation manner of the present application, and the protection scope of the present application is not limited thereto. Those skilled in the art may make changes or substitutions within the technical scope disclosed in this application, and these changes or substitutions shall all be within the protection scope of this application.

Claims (12)

1. A method for detecting the empty ratio of an adeno-associated viral vector, comprising the steps of:

determining the sedimentation coefficient value of the adeno-associated virus vector and the sedimentation distribution coefficient value corresponding to the sedimentation coefficient value by an analytical ultracentrifuge;

normalizing the sedimentation coefficient value and the sedimentation distribution coefficient value;

and obtaining the blank ratio by the ratio of each sedimentation distribution coefficient value to the sum of sedimentation distribution coefficient values.

2. The method for detecting the void fraction of adeno-associated virus vector according to claim 1, wherein the sedimentation curve measured by the analytical ultracentrifuge is used for obtaining sedimentation coefficients, molar masses and diffusion coefficient values, comprising the steps of:

using the c(s) analytical model of the SEDFIT software, which provides that all components in the adeno-associated viral vector solution have the same coefficient of friction ratio, the following formula is calculated:

f ₀ ＝6πηr ₀ ，

wherein f ₀ The friction coefficient of the polymer collapsed into compact pellets, eta is the viscosity of the solvent, r ₀ Is a hydrodynamic radius;

the following formula is deformed:

wherein M is molar mass, N _A In terms of mole number ρ _p Is the density of solution particles;

the method comprises the following steps:

wherein v is the specific volume of the solute;

according to Svedberg equation:

wherein s is a sedimentation coefficient value;

the molar mass is obtained:

substituting the sedimentation coefficient and the corresponding molar mass into the Svedberg equation to obtain a diffusion coefficient value:

wherein D is a diffusion coefficient value, R is a gas constant, T is an absolute temperature, and ρ is a solvent density.

3. The method for detecting the void fraction of an adeno-associated viral vector according to claim 1, wherein obtaining the sedimentation coefficient value and the sedimentation distribution coefficient value comprises the steps of:

processing the data measured by the analytical ultracentrifuge through SEDFIT software;

and integrating the processed data by integrating the data of each peak displayed by GUSSI software processing to obtain the sedimentation coefficient value and the sedimentation distribution coefficient value.

4. The method for detecting the empty ratio of adeno-associated virus vectors according to claim 1, wherein the method comprises the steps of: the reference substance of the adeno-associated viral vector is a preparation buffer solution stored in the adeno-associated viral vector virus liquid.

5. The method for detecting the empty ratio of adeno-associated virus vectors according to claim 4, wherein the method comprises the steps of: the formulation buffer includes water, saline solution, and cell culture medium.

6. The method for detecting the empty ratio of adeno-associated virus vectors according to claim 1, wherein the method comprises the steps of: the measurement temperature of the analytical ultracentrifuge is 15-25 ℃.

7. The method for detecting the empty ratio of adeno-associated virus vectors according to claim 1, wherein the method comprises the steps of: the absorbance of the adeno-associated viral vector is 0.3-1.5.

8. A void fraction detection system for adeno-associated viral vectors, comprising:

a measurement module for measuring the sedimentation coefficient value of the adeno-associated virus vector and the sedimentation distribution coefficient value corresponding to the sedimentation coefficient value by an analytical ultracentrifuge;

the normalization processing module is used for carrying out normalization processing on the sedimentation coefficient value and the sedimentation distribution coefficient value;

and the blank ratio calculation module is used for calculating the blank ratio through the ratio of each sedimentation distribution coefficient value to the sum of the sedimentation distribution coefficient values.

9. The void fraction detection system of claim 8, further comprising:

and the parameter setting module is used for setting parameters of the analytical ultracentrifuge.

10. A void ratio detection device for adeno-associated viral vectors comprises at least one processor; a memory coupled to the at least one processor, the memory storing executable instructions, characterized by: the executable instructions, when executed by the at least one processor, cause the steps of the method according to any one of claims 1 to 7 to be carried out.

11. A chip, characterized in that: comprising a processor for calling and running a computer program from a memory, such that a device on which the chip is mounted performs the steps of the method according to any of claims 1 to 7.

12. A computer-readable storage medium having a computer program stored thereon, characterized in that: the computer program, when executed by a processor, implements the steps of the method of any of the preceding claims 1 to 7.

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