Measurement properties of samples via isolated viral gene delivery vectors

Abstract

This disclosure describes apparatus, methods, and systems for measuring the properties of isolated viral gene delivery vector samples. In one embodiment, the method, system, and computer program product include: performing a set of logical operations on a set of analytical instruments to analyze the viral gene delivery vector sample, wherein the set includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors, thereby obtaining the capsid protein mass m of the sample. A The mass m of the modifier in the sample B The molar mass M of the modifier for the sample B The molar mass M of the capsid protein in the sample was received from the capsid protein molar mass data source. A ; Receive the injection volume v of the sample from the injection volume data source; and perform calculation of the total VGDV particle concentration C of the sample. A A set of logical operations.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Patent Application Serial No. 16 / 991,016, filed on August 11, 2020. Technical Field

[0003] This disclosure relates to samples, and more specifically, to the measurement properties of samples delivered via isolated viral gene delivery vectors. Background Technology

[0004] US2015005369A1 discloses a method for gene delivery using a capsid-modified recombinant adeno-associated virus (rAAV) vector. Exemplary methods include using a vector with altered affinity for heparin or heparin sulfate, and a vector lacking functional VP2 protein expression but still possessing full virulence, an expression system, and rAAV viral particles. Summary of the Invention

[0005] This disclosure describes computer-implemented methods, systems, and computer program products for measuring properties of isolated viral gene delivery vector (VGDV) samples. In one exemplary embodiment, the computer-implemented methods, systems, and computer program products include: (1) performing a set of logical operations by a computer system to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors, thereby obtaining the capsid protein mass m of the sample. A The mass m of the modifier in the sample B And the molar mass M of the modifier in the sample B (2) Receive the molar mass M of the capsid protein of the sample from the capsid protein molar mass data source. A (3) Receive the injection volume v of the sample from the injection volume data source, and (4) Calculate the total VGDV particle concentration C of the sample by the computer system using the following formula. A A set of logical operations:

[0006] ,

[0007] Where N is the Avogrado number. In one embodiment, the sample is a lentiviral vector sample. In one embodiment, the sample is an adenoviral vector sample. In one embodiment, the sample is an adeno-associated virus (AAV) sample. In one embodiment, the mass m of the modifier of the sample... B This refers to the nucleic acid mass of the sample. In one embodiment, M is the molar mass of the modifier in the sample. B It is the molar mass of nucleic acid in the sample. Attached Figure Description

[0008] Figure 1A A flowchart according to an exemplary embodiment is depicted.

[0009] Figure 1B A block diagram is depicted according to an exemplary embodiment.

[0010] Figure 1C A flowchart according to an exemplary embodiment is depicted.

[0011] Figure 1D A flowchart according to an exemplary embodiment is depicted.

[0012] Figure 1E A flowchart according to an exemplary embodiment is depicted.

[0013] Figure 1F A flowchart according to an exemplary embodiment is depicted.

[0014] Figure 1G A flowchart according to an exemplary embodiment is depicted.

[0015] Figure 2A A flowchart according to one embodiment is depicted.

[0016] Figure 2B A flowchart according to one embodiment is depicted.

[0017] Figure 3A An apparatus according to one embodiment is described.

[0018] Figure 3B An apparatus according to one embodiment is described.

[0019] Figure 3C An apparatus according to one embodiment is described.

[0020] Figure 3D An apparatus according to one embodiment is described.

[0021] Figure 3E A typical set of instruments is described.

[0022] Figure 4A A diagram according to one embodiment is depicted.

[0023] Figure 4B A diagram according to one embodiment is depicted.

[0024] Figure 4C A diagram according to one embodiment is depicted.

[0025] Figure 5 A diagram according to one embodiment is depicted.

[0026] Figure 6A A diagram according to one embodiment is depicted.

[0027] Figure 6B A diagram according to one embodiment is depicted.

[0028] Figure 6C A diagram according to one embodiment is depicted.

[0029] Figure 7A A diagram according to one embodiment is depicted.

[0030] Figure 7B A diagram according to one embodiment is depicted.

[0031] Figure 7C A diagram according to one embodiment is depicted.

[0032] Figure 8 A computer system according to an exemplary embodiment is described. Detailed Implementation

[0033] This disclosure describes computer-implemented methods, systems, and computer program products for measuring the properties of isolated viral gene delivery vector (VGDV) samples. In one exemplary embodiment, the computer-implemented methods, systems, and computer program products include: (1) performing a set of logical operations by a computer system to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors, thereby obtaining the capsid protein mass m of the sample. A The mass m of the modifier in the sample B And the molar mass M of the modifier in the sample B (2) Receive the molar mass M of the capsid protein of the sample from the capsid protein molar mass data source. A (3) Receive the injection volume v of the sample from the injection volume data source, and (4) The computer system performs a set of logical operations to calculate the total VGDV particle concentration C of the sample by the following formula. A :

[0034] ,

[0035] Where N is the Avogrado number. In one embodiment, the sample is a lentiviral vector sample. In one embodiment, the sample is an adenoviral vector sample. In one embodiment, the sample is an adeno-associated virus (AAV) sample. In one embodiment, the mass m of the modifier of the sample... BThis refers to the nucleic acid mass of the sample. In one embodiment, M is the molar mass of the modifier in the sample. B It is the molar mass of nucleic acid in the sample.

[0036] definition

[0037] Particles

[0038] The particles can be components of a liquid sample aliquots. Such particles can be molecules of different types and sizes, nanoparticles, virus-like particles, liposomes, emulsions, bacteria, and colloids. The size range of these particles can be from nanometers to micrometers.

[0039] Analysis of the types of macromolecules or particles in solution

[0040] Analysis of macromolecules or particle species in solution can be achieved by preparing samples in a suitable solvent and then injecting aliquots of the sample into a separation system, such as a liquid chromatography (LC) column or field flow fractionation (FFF) channel, where different particle species contained in the sample are separated into different components. Once separated, typically based on size, mass, or column affinity, the sample can be analyzed by light scattering, refractive index, UV absorption, electrophoretic mobility, and viscometer response.

[0041] Light scattering

[0042] Light scattering (LS) is a non-invasive technique for characterizing macromolecules and various particles in solution. Two common types of light scattering detection used to characterize macromolecules are static light scattering and dynamic light scattering.

[0043] Dynamic light scattering

[0044] Dynamic light scattering, also known as quasi-elastic light scattering (QELS) and photon correlation spectroscopy (PCS), involves measuring the time-varying fluctuations in the scattered light signal using a fast photodetector. DLS measurements determine the diffusion coefficient of molecules or particles, which can then be used to calculate their hydrodynamic radii.

[0045] Static light scattering

[0046] Static light scattering (SLS) encompasses various techniques, such as single-angle light scattering (SALS), two-angle light scattering (DALS), small-angle light scattering (LALS), and multi-angle light scattering (MALS). SLS experiments typically involve measuring the absolute intensity of light scattered from a sample in solution illuminated by a narrow beam of light. This measurement is commonly used for appropriate classes of particles / molecules to determine the size and structure of sample molecules or particles, and, when combined with knowledge of sample concentration, to determine the weight-average molar mass. Furthermore, the nonlinearity of the scattered light intensity as a function of sample concentration can be used to measure interparticle interactions and association.

[0047] Multi-angle light scattering

[0048] Multi-angle light scattering (MALS) is a type of light scattering (SLS) technique used to measure light scattered by a sample at multiple angles. It is used to determine the absolute molar mass and average size of molecules in a solution by detecting how they scatter light.

[0049] Collimated light from a laser source is most commonly used, in which case the technique can be called multi-angle laser scattering (MALLS). The term "multi-angle" refers to the detection of scattered light at different discrete angles, such as by measuring a single detector that moves within a range including selected specific angles or by an array of detectors fixed at a specific angular position.

[0050] MALS measurements require a set of auxiliary components. The most important of these is a collimated or focused beam (typically from a laser source that produces the collimated monochromatic beam) illuminating the sample area. The beam is usually plane-polarized, perpendicular to the measurement plane, but other polarizations can be used, especially when studying anisotropic particles. Another essential component is an optical cell for containing the sample under test. Alternatively, a cell containing means that allow measurement of a flowing sample can be used. If the scattering properties of individual particles are to be measured, a method must be provided to introduce such particles one at a time through the beam at points substantially equidistant from surrounding detectors.

[0051] While most MALS-based measurements are performed in a plane containing a set of detectors typically equidistant from the sample at the center through which the illumination beam passes, three-dimensional versions have also been developed where the detectors are located on the surface of a sphere and the sample is controlled to pass through its center, where it intersects the path of the incident beam traveling along the diameter of the sphere. MALS techniques typically collect multiplexed data sequentially from the outputs of a set of discrete detectors. MALS light scattering photometers generally have multiple detectors.

[0052] It may be necessary to normalize the signals captured by the photodetectors of the MALS detector at each angle because different detectors in the MALS detector may (i) have slightly different quantum efficiencies and different gains, and (ii) may account for different geometric scattering volumes. Without normalizing these differences, the results of the MALS detector may be meaningless and incorrectly weighted for different detector angles.

[0053] Concentration detector

[0054] Differential refractive index detector

[0055] A differential refractive index detector (dRI), or differential refractometer, or refractive index detector (RI or RID), is a detector that measures the refractive index of an analyte relative to a solvent. They are commonly used in high-performance liquid chromatography (HPLC) and size exclusion chromatography. dRIs are considered universal detectors because they can detect any substance with a refractive index different from that of the solvent, but they have relatively low sensitivity. When light leaves one material and enters another, it bends or refracts. The refractive index of a material is a measure of how much light bends as it enters.

[0056] A differential refractive index detector (dRI) comprises a flow cell consisting of two parts: one for the sample and one for a reference solvent. The dRI measures the refractive indices of the two components. When only the solvent passes through the sample component, the refractive indices of both components are measured to be the same. However, when the analyte passes through the flow cell, the two refractive indices are measured to be different. The difference is displayed as peaks in the chromatogram. Differential refractive index detectors are commonly used for the analysis of polymer samples in size exclusion chromatography. The dRI can output a concentration detector signal value corresponding to the sample concentration.

[0057] UV-Vis Spectroscopy

[0058] Ultraviolet-visible spectroscopy, or ultraviolet-visible spectrophotometry (UV-Vis or UV / Vis), refers to the absorption or reflectance spectra in the ultraviolet-visible spectral region. UV-Vis detectors / UV-Vis spectrophotometers use light in the visible and adjacent ranges, where absorption or reflectance directly affects the perceived color of the chemical substance involved. In this region of the electromagnetic spectrum, atoms and molecules undergo electronic transitions. This absorption spectroscopy measures transitions from the ground state to an excited state. A UV-Vis detector / UV-Vis spectrophotometer measures the light intensity (I) passing through the sample and compares it to the light intensity (I0) before passing through the sample. o ) are compared, where I / I o The ratio of absorbance to light intensity is called transmittance, usually expressed as a percentage (%T). Absorbance A is based on transmittance according to the following formula:

[0059] A = - log (%T / 100%).

[0060] The UV-Vis spectrophotometer can also be configured to measure reflectance, wherein the spectrophotometer measures the intensity (I) of light reflected from the sample and compares it with the intensity (I0) of light reflected from a reference material. o The ratio I / I is compared. o This is called reflectance, and is usually expressed as a percentage (%R). A UV absorption detector outputs a concentration detector signal value corresponding to the sample concentration.

[0061] Adeno-associated virus

[0062] Adeno-associated virus (AAV) is a small virus (~20 nm) belonging to the Parvoviridae family that can infect humans but is believed not to cause any disease. Due to its small size, mild immune response, and ability to stably integrate its genome into the host cell genome at a specific site (AAVS 1 on human chromosome 19), AAV has become an attractive vector for gene therapy. With the recent FDA approval of Zolgesma® for the treatment of spinal muscular dystrophy and several promising ongoing clinical trials, such as trial number NCT00516477 (Clinicaltrials.gov), the AAV manufacturing process requires robust, reliable, and easily implemented characterization methods to meet regulatory requirements imposed by the FDA and other regulatory agencies. However, the characterization of viral vectors remains a challenge, and new methods must be developed to ensure safe and high-quality AAV vectors to advance AAV-related clinical research. Furthermore, the FDA has recently developed two reference standard materials (RSMs), recombinant AAV serotypes 2 and 8, which can be used as benchmark tools to demonstrate that characterization methods are properly controlled and for identifying internal reference materials. In establishing these RSMs, it was noted that there were significant differences in the determination of capsid particles and vector genomes among different institutions, which further underscores the need for reliable methods to determine the titers of AAV vectors in preclinical and clinical studies as the field develops so rapidly.

[0063] AAV characterization typically involves several distinct stages: particle titer, vector genome titer, transduction titer, infectivity titer, and determination of purity and identity. Particle titer quantification usually involves an ELISA (enzyme-linked immunosorbent assay) to detect the presence of a ligand (protein) in solution using an antibody against the target protein. Because this test requires a specialized laboratory, results typically take 24 hours to several weeks. The next step is to quantify the amount of viral genome. This is usually done using qPCR (quantitative polymerase chain reaction) after the viral capsid has lysed and the sample has been lost.

[0064] The purity and identity of the obtained AAV were assessed by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). The stoichiometry and size of the obtained viral capsid protein bands were evaluated. However, this is a relative method and requires the use of standards. The identity of the gene vector was determined by observing the electrophoretic band patterns and comparing them with a positive control (also a relevant technique).

[0065] Therefore, it is crucial to have robust and reproducible methods that can be easily implemented in QC during manufacturing. SEC-MALS allows for rapid sample analysis with run times under 30 minutes and can be used to determine key quality properties of AAV-based gene therapy agents, such as viral capsid concentration, the ratio of filled to unfilled particles, the absolute molar mass of proteins, and genomic composition. Properties of viral gene delivery vector (VGDV) samples need to be measured by separation.

[0066] refer to Figure 1A In one exemplary embodiment, a computer-implemented method, system, and computer program product are configured to perform: operation 110, by which a computer system performs a set of logical operations to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors, thereby obtaining the capsid protein mass m of the sample. A The mass m of the modifier in the sample B The molar mass M of the modifier for the sample B The molar mass M of the capsid protein in the sample was received from the capsid protein molar mass data source. A Operation 112; operation 114: receiving the injection volume v of the sample from the injection volume data source; and operation 116: a set of logical operations performed by the computer system, which calculate the total VGDV particle concentration C of the sample via the following formula. A :

[0067] .

[0068] Where N is the Avogrado number.

[0069] In an exemplary embodiment, the computer system is a standalone computer system, such as... Figure 8 The computer system shown is 800. It is a distributed computer network in which at least some of the computers are, for example, Figure 8 The computer system shown is a computer system 800, or a cloud computing node server, such as Figure 8 The computer system 800 shown. In one embodiment, the computer system is as follows: Figure 8The computer system 800 shown performs measurement of the properties of a viral gene delivery vector (VGDV) sample via a separation script or computer software application that implements at least the operations of method 100. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least the operations of method 100. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 100. In one embodiment, the computer system is the processor of an analytical instrument that performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 100.

[0070] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 110, 112, 114, and 116. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 110, 112, 114, and 116. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs the measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 110, 112, 114 and 116.

[0071] refer to Figure 1B In an exemplary embodiment, the computer-implemented method, system, and computer program product include an analyzer 120, a receiver 122, and a calculator 124. In one embodiment, the analyzer 120 is configured to perform a set of logical operations on a set of analytical instruments 130 to analyze a viral gene delivery vector (VGDV) sample 132, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors, thereby obtaining the capsid protein mass 140 m from the sample. A The mass of the modifier in the sample was 142 m. B The molar mass of the modifier in the sample was 144 M. B In one embodiment, the analyzer 120 includes a computer system that performs operation 110, such as... Figure 8 The computer system 800 is shown. In one embodiment, the analyzer 120 includes a computer system that performs operation 110, such as... Figure 8 The computer system / server 812 shown. In one embodiment, the analyzer 120 includes a computer system that performs operation 110, such as... Figure 8 The processing unit 816 is shown. In one embodiment, the analyzer 120 is implemented in a computer system such as Figure 8 Computer software executing on the computer system 800 shown causes the computer system to perform operation 110. In one embodiment, the analyzer 120 is implemented in a computer system such as Figure 8 The computer software executing on the computer system / server 812 shown causes the computer system to perform operation 110. In one embodiment, the analyzer 120 is implemented on the computer system such as Figure 8 The computer software executing on the processing unit 816 causes the computer system to perform operation 110. In one embodiment, the analyzer 120 performs operation 110 as computer software executing on the processor of the analyzer 120.

[0072] In one embodiment, receiver 122 is configured to receive the molar mass 136 M of the capsid protein of a sample from capsid protein molar mass data source 134. A In one embodiment, receiver 122 includes a computer system that performs operation 112, such as... Figure 8 The computer system 800 is shown. In one embodiment, receiver 122 includes a computer system performing operation 112, such as... Figure 8 The computer system / server 812 shown. In one embodiment, receiver 122 includes a computer system performing operation 112, such as... Figure 8 The processing unit 816 is shown. In one embodiment, the receiver 122 is implemented in a computer system such as Figure 8 Computer software executing on the computer system 800 shown causes the computer system to perform operation 112. In one embodiment, receiver 122 is implemented in the computer system such as Figure 8 Computer software executing on the computer system / server 812 shown causes the computer system to perform operation 112. In one embodiment, receiver 122 is implemented in the computer system such as Figure 8 The computer software executing on the processing unit 816 shown causes the computer system to perform operation 112. In one embodiment, the receiver 122 performs operation 112 as computer software executing on the processor of the receiver 122.

[0073] In one embodiment, receiver 122 is configured to receive an injection volume 139v of a sample from injection volume data source 138. In one embodiment, receiver 122 includes a computer system that performs operation 114, such as... Figure 8 The computer system 800 is shown. In one embodiment, receiver 122 includes a computer system performing operation 114, such as... Figure 8 The computer system / server 812 is shown. In one embodiment, the receiver 122 includes a computer system performing operation 114, such as... Figure 8 The processing unit 816 is shown. In one embodiment, the receiver 122 is implemented in a computer system such as Figure 8 The computer software executing on the computer system 800 shown causes the computer system to perform operation 114. In one embodiment, the receiver 122 is implemented as computer software executing on the computer system, such as... Figure 8 The computer system / server 812 shown causes the computer system to perform operation 114. In one embodiment, the receiver 122 is implemented in the computer system such as Figure 8 The computer software executing on the processing unit 816 shown causes the computer system to perform operation 114. In one embodiment, the receiver 122 performs operation 114 as computer software executing on the processor of the receiver 122.

[0074] In one embodiment, calculator 124 is configured to perform a calculation of the total VGDV particle concentration 146 C of the sample via the following formula. A A set of logical operations:

[0075] ,

[0076] Where N is an Avogrado number. In one embodiment, calculator 124 includes a computer system that performs operation 116, such as... Figure 8 The computer system 800 is shown. In one embodiment, the calculator 124 includes a computer system that performs operation 116, such as... Figure 8 The computer system / server 812 is shown. In one embodiment, the calculator 124 includes a computer system that performs operation 116, such as... Figure 8 The processing unit 816 is shown. In one embodiment, the calculator 124 is implemented in a computer system such as Figure 8 Computer software executing on the computer system 800 shown causes the computer system to perform operation 116. In one embodiment, the calculator 124 is implemented as a computer system such as Figure 8The computer software executing on the computer system / server 812 shown causes the computer system to perform operation 116. In one embodiment, the calculator 124 is implemented as a computer system (such as...) Figure 8 The computer software executing on the processing unit 816 (shown) causes the computer system to perform operation 116. In one embodiment, the calculator 124 performs operation 116 as computer software executing on the processor of the calculator 124.

[0077] instrument

[0078] Separation instruments

[0079] In one embodiment, at least one separation instrument includes at least one of size exclusion chromatography (SEC) unit, field flow fractionation (FFF) unit, and ion exchange chromatography (IEX) unit. In another embodiment, at least one separation instrument is at least one of SEC unit, FFF unit, and IEX unit.

[0080] Static light scattering instrument

[0081] In one embodiment, at least one static light scattering (SLS) instrument includes a multi-angle light scattering (MALS) instrument. In another embodiment, at least one static light scattering (SLS) instrument is a MALS instrument.

[0082] Concentration detector

[0083] UV-UV

[0084] In one embodiment, at least two concentration detectors include a first ultraviolet absorbance (UV) detector with a first wavelength λ1 and a second ultraviolet absorbance (UV) detector with a second wavelength λ2. In another embodiment, at least two concentration detectors are a first UV detector with a first wavelength λ1 and a second UV detector with a second wavelength λ2. In a particular embodiment, the first wavelength λ1 is 260 nm and the second wavelength λ2 is 280 nm.

[0085] UV-dRI

[0086] In one embodiment, at least two concentration detectors include an ultraviolet absorbance (UV) detector and a differential refractive index (dRI) detector with a wavelength of λ. In another embodiment, at least two concentration detectors are a UV detector and a dRI detector with a wavelength of λ. In a particular embodiment, the wavelength λ is one of 260 nm and 280 nm.

[0087] UV-FLD

[0088] In one embodiment, at least two concentration detectors include an ultraviolet absorbance (UV) detector and a fluorescence detector (FLD) with a wavelength of λ. In one embodiment, at least two concentration detectors are a UV detector and an FLD with a wavelength of λ. In a particular embodiment, the wavelength λ is one of 260 nm and 280 nm.

[0089] dRI-FLD

[0090] In one embodiment, at least two concentration detectors include a differential refractive index (dRI) detector and a fluorescence detector (FLD). In another embodiment, at least two concentration detectors are a dRI detector and an FLD.

[0091] Detector selection

[0092] The use of different concentration detectors may depend on the initial sample mass, concentration, and total volume available for analysis. The sensitivity of this method depends on... Figure 3A The factors summarized in the table below.

[0093] Typically, each combination of concentration detectors can be used as follows:

[0094] dRI-FLD – Used only when the sample has a fluorescent tag with known excitation and emission wavelengths;

[0095] UV-FLD - for sample concentrations of ~10¹⁰ particles / mL (approximately 10¹⁰ particles / mL).

[0096] UV-UV – for sample concentrations of ~10¹¹ particles / mL (approximately 10¹¹ particles / mL); and

[0097] UV-dRI is used for sample concentrations higher than 10¹² particles / mL.

[0098] A set of instruments

[0099] refer to Figure 3B In one embodiment, at least one separation instrument 310 is connected to a first concentration detector 312, wherein the first concentration detector 312 is connected to at least one SLS instrument 314, wherein at least one SLS instrument 314 is connected to a second concentration detector 316. (Reference) Figure 3C In one embodiment, at least one separation instrument 320 is connected to a first concentration detector 322, wherein the first concentration detector 322 is connected to a second concentration detector 324, wherein the second concentration detector 324 is connected to at least one SLS instrument 326. (Reference) Figure 3DIn one embodiment, at least one separation instrument 330 is connected to at least one SLS instrument 332, wherein at least one SLS instrument 332 is connected to a first concentration detector 334, wherein the first concentration detector 334 is connected to a second concentration detector 336. For example, Figure 3E A typical set of instruments is described.

[0100] Calculate the molar mass of capsid protein

[0101] In another embodiment, the computer-implemented method, system, and computer program product further include: (a) a set of logical operations performed by the computer system on the sample to obtain the molar mass M of the capsid protein of the sample. A (a) and (b) the molar mass M of the capsid protein of the sample A The data is stored in a capsid protein molar mass data source. In a further embodiment, the method, system, and computer program product further include: (a) executing a set of logical operations on the sample on analysis group 130 by a computer system to obtain the capsid protein molar mass 136 M of the sample. A (a) and (b) the molar mass of the capsid protein in the sample was 136 M. A It is stored in capsid protein molar mass data source 134.

[0102] Analytical Samples

[0103] In one embodiment, the analysis includes analyzing samples on the group using analytical techniques, wherein the analytical techniques are one of viral vector analysis, protein conjugate analysis, and copolymer composition analysis. In one embodiment, analytical operation 110 includes analyzing sample 132 on group 130 using analytical techniques, wherein the analytical techniques are one of viral vector analysis, protein conjugate analysis, and copolymer composition analysis.

[0104] VGDV concentration

[0105] In a further embodiment, the computer-implemented method, system, and computer program product further include: (a) receiving, by the computer system, the molar mass M of the fully modified agent within the full VGDV sample from a fully modified agent molar mass data source. Full (b) The total VGDV concentration C of the total VGDV sample is calculated by a computer system using the following formula. Full A set of logical operations:

[0106] ,and

[0107] (c) The empty VGDV concentration C of the full VGDV sample is calculated by a computer system using the following formula. Empty A set of logical operations:

[0108] .

[0109] refer to Figure 2A The computer-implemented method, system, and computer program product are further configured to execute: receiving, by the computer system, the molar mass M of the total modifier within the total VGDV sample from a total modifier molar mass data source. Full Operation 210; Operation 212, performed by the computer system, is a set of logical operations that calculate the total VGDV concentration C of the total VGDV sample via the following formula. Full :

[0110] ,

[0111] Operation 214, which is performed by a computer system, consists of a set of logical operations that calculate the empty VGDV concentration C of the full VGDV sample via the following formula. Empty :

[0112] .

[0113] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement of properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least the operations of method 200. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of the properties of a viral gene delivery vector (VGDV) sample via a separation script or computer software application that implements at least the operations of method 200. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 200. In one embodiment, the computer system is the processor of an analytical instrument that performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 200.

[0114] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that executes at least operations 210, 212, and 214. In one embodiment, the computer system is as follows: Figure 8The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 210, 212, and 214. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs measurement of the properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 210, 212 and 214.

[0115] In another embodiment, the method, system, and computer program product further include: (a) a set of logical operations performed by the computer system to analyze the whole VGDV sample on a group basis, thereby obtaining the molar mass M of the total modifier within the whole VGDV sample. Full and (b) the full VGDV sample M Full The molar mass M of the internal modifier Full The data is stored in a data source of the total modifier molar mass. In a further embodiment, the method, system, and computer program product further include: (a) a set of logical operations performed by the computer system to analyze the whole VGDV sample on group 130 to obtain the molar mass M of the total modifier in the whole VGDV sample. Full (b) The molar mass M of the total modifier in the full VGDV sample Full It is stored in a data source containing the total molar mass of the modifier. In one embodiment, the modifier is a nucleic acid.

[0116] Peak particle concentration of total VGDV

[0117] In another embodiment, the method, system, and computer program product further include: (a) a set of logical operations performed by a computer system to analyze the entire VGDV signal region of the sample on a group and to analyze the aggregated peak region of the sample on a group, thereby obtaining the entire peak protein mass m of the VGDV of the sample corresponding to the entire VGDV signal region of the sample. A,ent The mass m of the modifier corresponding to the entire VGDV signal region of the sample. B,ent The molar mass M of the protein at the entire peak of VGDV corresponding to the entire VGDV signal region of the sample. A,ent The molar mass M of the modifier corresponding to the entire VGDV signal region of the sample. B,ent The protein mass m of the VGDV aggregation peak corresponding to the aggregation peak region of the sample. A,agg The mass m of the VGDV aggregation peak modifier corresponding to the aggregation peak region of the sample. B,agg The molar mass M of the protein at the VGDV aggregation peak corresponding to the aggregation peak region of the sample.A,agg The molar mass M of the VGDV aggregation peak modifier corresponding to the aggregation peak region of the sample. B,agg (b) A set of logical operations is performed by the computer system, which calculates the total VGDV peak particle concentration C of the sample corresponding to the entire VGDV signal region of the sample via the following formula. A,ent :

[0118] ,and

[0119] (c) The computer system performs a set of logical operations to calculate the total VGDV peak particle concentration C of the sample corresponding to the peak aggregation region of the sample via the following formula. A,agg :

[0120] .

[0121] refer to Figure 2B The computer-implemented method, system, and computer program product are further configured to perform operation 222, which is a set of logical operations executed by the computer system. This set of logical operations analyzes the entire VGDV signal region of the sample on the set and analyzes the aggregated peak region of the sample on the set, thereby obtaining the entire peak protein mass m of the sample's VGDV corresponding to the entire VGDV signal region of the sample. A,ent The mass m of the modifier corresponding to the entire VGDV signal region of the sample. B,ent The molar mass M of the protein at the entire peak of VGDV corresponding to the entire VGDV signal region of the sample. A,ent The molar mass M of the modifier corresponding to the entire VGDV signal region of the sample. B,ent The protein mass m of the VGDV aggregation peak corresponding to the aggregation peak region of the sample. A,agg The mass m of the VGDV aggregation peak modifier of the sample corresponding to the aggregation peak region. B,agg The molar mass M of the protein at the VGDV aggregation peak corresponding to the aggregation peak region of the sample. A,agg The molar mass M of the VGDV aggregation peak modifier corresponding to the aggregation peak region of the sample. B,agg Operation 224, performed by the computer system, is a set of logical operations that calculate the total VGDV peak particle concentration C of the sample corresponding to the entire VGDV signal region of the sample via the following formula. A,ent :

[0122] ,and

[0123] Operation 226, performed by a computer system, involves a set of logical operations that calculate the total VGDV peak particle concentration C of the sample corresponding to the peak aggregation region, using the following formula. A,agg :

[0124] .

[0125] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that executes at least the operations of method 220. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of the properties of a viral gene delivery vector (VGDV) sample via a separation script or computer software application that implements at least the operations of method 220. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 220. In one embodiment, the computer system is the processor of an analytical instrument that performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 220.

[0126] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that executes at least operations 222, 224, and 226. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 222, 224, and 226. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs measurement of the properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 222, 224 and 226.

[0127] In another embodiment, the method, system, and computer program product further include: (a) a set of logical operations performed by a computer system to analyze the entire VGDV signal region of the whole viral gene delivery vector sample on the group and to analyze the aggregated peak region of the whole sample on the group, thereby obtaining the entire peak molar mass M of the VGDV of the whole modifier in the whole sample corresponding to the entire VGDV signal region of the whole sample.Full,ent And the molar mass M of the VGDV aggregation peak of the entire modifier within the entire sample corresponding to the aggregation peak region of the entire sample. Full,agg (b) A set of logical operations is performed by the computer system, which calculates the total peak VGDV concentration C of the whole sample corresponding to the entire VGDV signal region of the VGDV sample via the following formula. Full,ent :

[0128] ,

[0129] (c) A set of logical operations is performed by the computer system, which calculates the total VGDV concentration C of the whole sample corresponding to the aggregation peak region of the whole sample via the following formula. Full,agg :

[0130] ,

[0131] (d) A set of logical operations is executed by the computer system, which calculates the peak empty VGDV concentration C of the whole sample corresponding to the entire VGDV signal region of the whole sample via the following formula. Empty,ent :

[0132] ,and

[0133] (e) A set of logical operations is performed by the computer system, which calculates the empty VGDV concentration C of the full sample corresponding to the aggregation peak region of the full sample via the following formula. Empty,agg :

[0134] .

[0135] In another embodiment, the method, system, and computer program product further include: (a) a set of logical operations performed by the computer system to analyze the entire VGDV signal region of the whole viral gene delivery vector sample on group 130 and to analyze the aggregation peak region of the whole sample on group 130, thereby obtaining: the entire peak molar mass M of the VGDV of the whole modifier within the whole sample corresponding to the entire VGDV signal region of the whole sample. Full, ent And the molar mass M of the VGDV aggregation peak of the entire modifier within the entire sample corresponding to the aggregation peak region of the entire sample. Full,agg (b) A set of logical operations is performed by the computer system, which calculates the total peak VGDV concentration C of the whole sample corresponding to the entire VGDV signal region of the VGDV sample via the following formula. Full,ent :

[0136] ,

[0137] (c) A set of logical operations is performed by the computer system, which calculates the total VGDV concentration C of the whole sample corresponding to the aggregation peak region of the whole sample via the following formula. Full,agg :

[0138] ,

[0139] (d) A set of logical operations is executed by the computer system, which calculates the peak empty VGDV concentration C of the whole sample corresponding to the entire VGDV signal region of the whole sample via the following formula. Empty,ent :

[0140] ,and

[0141] (e) A set of logical operations is performed by the computer system, which calculates the empty VGDV concentration C of the full sample corresponding to the aggregation peak region of the full sample via the following formula. Empty,agg :

[0142] .

[0143] Approximate total VGDV particle concentration

[0144] In one exemplary embodiment, the computer-implemented method, system, and computer program product include: (1) a set of logical operations performed by a computer system to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least one concentration detector, thereby obtaining the mass m of the modifier of the sample. B Molar mass M of the modifier in the sample B (2) The sample’s capsid protein mass m is calculated by a computer system using at least one UV extinction coefficient relative to at least one refractive index increment value from at least one concentration detector. A And the molar mass M of the capsid protein of the sample A (3) Receive the injection volume v of the sample from the injection volume data source, and (4) The computer system performs a set of logical operations that calculate the total VGDV particle concentration C of the sample via the following formula. A :

[0145] ,

[0146] Where N is the Avogrado number. In one embodiment, at least one concentration detector is a dRI detector.

[0147] refer to Figure 1CIn an exemplary embodiment, the computer-implemented method, system, and computer program product are configured to perform: operation 152, which involves a computer system performing a set of logical operations to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least one concentration detector, thereby obtaining the mass m of the modifier for the sample. B Molar mass M of the modifier in the sample B and at least one UV extinction coefficient of the sample; operation 154, performed by a computer system, is a set of logical operations relative to at least one refractive index increment value from at least one concentration detector to calculate the capsid protein mass m of the sample. A And the molar mass M of the capsid protein of the sample A ; Operation 156 of receiving the injection volume v of the sample from the injection volume data source, and (4) Operation 158 of performing a set of logical operations by the computer system, which calculates the total VGDV particle concentration C of the sample via the following formula. A ,

[0148] ,

[0149] Where N is the Avogrado number.

[0150] In an exemplary embodiment, the computer system is: a standalone computer system, such as Figure 8 The computer system 800 shown; a distributed computer network, wherein at least some of the computers are such as Figure 8 The computer system shown is a computer system of computer system 800; or a cloud computing node server, such as Figure 8 The computer system 800 shown. In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement of the properties of a viral gene delivery vector (VGDV) sample via a separation script or computer software application that implements at least the operations of method 150. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least the operations of method 150. In one embodiment, the computer system is as follows: Figure 8The processing unit 816 shown performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 150. In one embodiment, the computer system is the processor of an analytical instrument that performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 150.

[0151] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 152, 154, 156, and 158. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 152, 154, 156, and 158. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 152, 154, 156 and 158.

[0152] Total VGDV particle concentration using a UV detector

[0153] In one exemplary embodiment, the computer-implemented method, system, and computer program product include: (1) a set of logical operations performed by the computer system to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and (2) a set of logical operations performed by the computer system relative to an ultraviolet absorbance value A collected from the sample at a first wavelength λ1. λ1 The ultraviolet absorbance value A collected from the sample at the second wavelength λ2 λ2 The extinction coefficient of the protein at the first wavelength λ1 The extinction coefficient of the protein at the second wavelength λ2 The extinction coefficient of the modifier in the sample at the first wavelength λ1 and the extinction coefficient of the modifier in the sample at the second wavelength λ2 To calculate the mass fraction X of protein in the sample A (3) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The extinction coefficient of the protein at the first wavelength And the extinction coefficient of the modifier in the sample at the first wavelength To calculate the extinction coefficient of the sample at the first wavelength. (4) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The extinction coefficient of the protein at the second wavelength and the extinction coefficient of the modifier in the sample at the second wavelength To calculate the extinction coefficient of the sample at the second wavelength. (5) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample. VGDV (6) A set of logical operations is executed by the computer system relative to the ultraviolet absorbance value A collected from the sample at wavelength λ. λ The mass fraction of protein in the sample, X A The extinction coefficient of the protein at wavelength λ The extinction coefficient of the modifier in the sample at wavelength λ To calculate the total protein mass m A and the total mass of the modifier m B Where wavelength λ is one of the first wavelength λ1 and the second wavelength λ2, and (7) is a set of logical operations performed by the computer system, which calculate the total VGDV particle concentration of the sample via the following formula:

[0154] ,

[0155] Where N is the Avogrado number, and M A This refers to the molar mass of the capsid protein in the sample, derived from a data source. In one embodiment, at least two concentration detectors include a first ultraviolet absorbance (UV) detector at a first wavelength λ1 and a second ultraviolet absorbance (UV) detector at a second wavelength λ2. In another embodiment, at least two concentration detectors are a first ultraviolet absorbance (UV) detector at a first wavelength λ1 and a second ultraviolet absorbance (UV) detector at a second wavelength λ2.

[0156] refer to Figure 1D and Figure 1EIn an exemplary embodiment, the computer-implemented method, system, and computer program product are configured to perform: operation 161, by which the computer system performs a set of logical operations to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and operation 162, by which the computer system performs a set of logical operations relative to an ultraviolet absorbance value A collected from the sample at a first wavelength λ1. λ1 The ultraviolet absorbance value A collected from the sample at the second wavelength λ2 λ2 The extinction coefficient of the protein at the first wavelength λ1 The extinction coefficient of the protein at the second wavelength λ2 The extinction coefficient of the modifier in the sample at the first wavelength λ1 and the extinction coefficient of the modifier in the sample at the second wavelength λ2 To calculate the mass fraction X of protein in the sample A Operation 163 is a set of logical operations performed by a computer system, which are relative to the mass fraction X of protein in the sample. A The extinction coefficient of the protein at the first wavelength And the extinction coefficient of the modifier in the sample at the first wavelength To calculate the extinction coefficient of the sample at the first wavelength. Operation 164 is a set of logical operations performed by a computer system, which are relative to the mass fraction X of protein in the sample. A The extinction coefficient of the protein at the second wavelength and the extinction coefficient of the modifier in the sample at the second wavelength To calculate the extinction coefficient of the sample at the second wavelength. Operation 165 is a set of logical operations performed by a computer system, which are relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample. VGDV Operation 166, performed by a computer system, is a set of logical operations relative to the ultraviolet absorbance value A collected from the sample at wavelength λ. λ The mass fraction of protein in the sample, X A The extinction coefficient of the protein at wavelength λ The extinction coefficient of the modifier in the sample at wavelength λ To calculate the total protein mass mA and the total mass of the modifier m B Where wavelength λ is one of a first wavelength λ1 and a second wavelength λ2; and operation 167, which is a set of logical operations performed by a computer system, calculates the total VGDV particle concentration C of the sample via the following formula. A :

[0157] ,

[0158] Where N is the Avogrado number, and where M A It is the molar mass of the capsid protein in the sample, derived from a data source of capsid protein molar mass.

[0159] In an exemplary embodiment, the computer system is: a standalone computer system, such as Figure 8 The computer system 800 shown; a distributed computer network, wherein at least some of the computers are such as Figure 8 The computer system shown is a computer system of computer system 800; or a cloud computing node server, such as Figure 8 The computer system 800 shown. In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement of the properties of a viral gene delivery vector (VGDV) sample via a separation script or computer software application that implements at least the operations of method 160. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least the operations of method 160. In one embodiment, the computer system is... Figure 8 The processing unit 816 shown performs the measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that performs at least the operations of method 160. In one embodiment, the computer system is the processor of an analytical instrument that performs the measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that performs at least the operations of method 160.

[0160] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 161, 162, 163, 164, 165, 166, and 167. In one embodiment, the computer system is as follows: Figure 8The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 161, 162, 163, 164, 165, 166, and 167. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 161, 162, 163, 164, 165, 166 and 167.

[0161] In one embodiment, the mass fraction X of protein in the sample is calculated. A This includes calculating the protein mass fraction X in the sample using the following formula. A :

[0162] ,

[0163] Calculate the extinction coefficient of the sample at the first wavelength. This includes calculating the extinction coefficient of the sample at the first wavelength using the following formula. :

[0164] ,

[0165] Calculate the extinction coefficient of the sample at the second wavelength. This includes calculating the extinction coefficient of the sample at the second wavelength using the following formula. :

[0166] ,

[0167] Calculate the refractive index increment (dn / dc) of the sample. VGDV This includes calculating the refractive index increment (dn / dc) of the sample using the following formula. VGDV :

[0168] ,

[0169] Calculate the total mass m of the protein A This includes calculating the total mass m of protein using the following formula. A :

[0170] ,

[0171] And calculate the total mass m of the modifier. B The total mass m of the modifier is calculated using the following formula. B :

[0172] .

[0173] In one embodiment, the mass fraction X of protein in the sample is calculated. A The calculation operation 162 includes calculating the mass fraction X of protein in the sample via the following formula. A :

[0174]

[0175] Calculate the extinction coefficient of the sample at the first wavelength. The calculation operation 163 includes calculating the extinction coefficient of the sample at the first wavelength via the following formula. :

[0176] ,

[0177] Calculate the extinction coefficient of the sample at the second wavelength. The calculation operation 164 includes calculating the extinction coefficient of the sample at the second wavelength via the following formula. :

[0178] ,

[0179] Calculate the refractive index increment (dn / dc) of the sample. VGDV The calculation operation 165 includes calculating the refractive index increment (dn / dc) of the sample via the following formula. VGDV :

[0180] ,

[0181] Calculate the total mass m of the protein A The computational operation 166 includes calculating the total mass m of the protein via the following formula. A :

[0182] ,

[0183] And calculate the total mass m of the modifier. B The calculation operation 167 includes calculating the total mass m of the modifier via the following formula. B :

[0184] .

[0185] In one embodiment, the first wavelength λ1 is 260 nm and the second wavelength λ2 is 280 nm. In one embodiment, the modifier is a nucleic acid.

[0186] Total VGDV particle concentration using UV detector and dRI detector

[0187] In one exemplary embodiment, the computer-implemented method, system, and computer program product include: (1) a set of logical operations performed by the computer system to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and (2) a set of logical operations performed by the computer system relative to an ultraviolet absorbance value A collected from the sample at wavelength λ. λ The refractive index coefficient (dn / dc) of the modifier in the sample. B The differential refractive index dRI of the solution containing the sample, and the extinction coefficient of the modifier at wavelength λ. The extinction coefficient of proteins at a given wavelength The refractive index coefficient of proteins (dn / dc) A To calculate the protein mass fraction X in the sample A (3) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The extinction coefficient of the protein at the first wavelength and the extinction coefficient of the modifier in the sample at wavelength To calculate the extinction coefficient of the sample at the specified wavelength. (4) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample. VGDV (5) A set of logical operations is executed by the computer system, which are relative to the differential refractive index dRI of the solution containing the sample and the mass fraction X of the protein in the sample. A The refractive index coefficient of proteins (dn / dc) A And the refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the total mass m of the protein A The total mass m of the modifier B (6) A set of logical operations is performed by the computer system to calculate the total VGDV particle concentration C of the sample via the following formula. A :

[0188] ,

[0189] Where N is the Avogrado number, and where M AThis refers to the molar mass of the capsid protein in the sample, derived from a data source. In one embodiment, at least two concentration detectors include a UV absorbance detector and a differential refractive index (dRI) detector at wavelength λ. In another embodiment, at least two concentration detectors are a UV detector and a dRI detector at wavelength λ.

[0190] refer to Figure 1F and Figure 1G In one exemplary embodiment, a computer-implemented method, system, and computer program product are configured to perform: operation 171, by which a computer system performs a set of logical operations to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and operation 172, by which the computer system performs a set of logical operations relative to an ultraviolet absorbance value A collected from the sample at wavelength λ. λ The refractive index coefficient (dn / dc) of the modifier in the sample. B The differential refractive index dRI of the solution containing the sample, and the extinction coefficient of the modifier at wavelength λ. The extinction coefficient of the protein at wavelength λ The refractive index coefficient of proteins (dn / dc) A To calculate the protein mass fraction X in the sample A Operation 173 is a set of logical operations performed by a computer system, which are relative to the mass fraction X of protein in the sample. A The extinction coefficient of proteins at a given wavelength and the extinction coefficient of the modifier in the sample at wavelength To calculate the extinction coefficient of the sample at the specified wavelength. Operation 174 is a set of logical operations performed by a computer system, which are relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample. VGDV Operation 175 is a set of logical operations performed by a computer system, relative to the differential refractive index dRI of the solution containing the sample and the mass fraction X of the protein in the sample. A The refractive index coefficient of proteins (dn / dc) A And the refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the total mass m of the protein A The total mass m of the modifierB Operation 176, performed by a computer system, involves a set of logical operations that calculate the total VGDV particle concentration C of the sample using the following formula. A :

[0191] ,

[0192] Where N is the Avogrado number, and M A It is the molar mass of the capsid protein in the sample, derived from a data source of capsid protein molar mass.

[0193] In an exemplary embodiment, the computer system is: a standalone computer system, such as Figure 8 The computer system 800 shown; a distributed computer network, wherein at least some of the computers are such as Figure 8 The computer system shown is a computer system of computer system 800; or a cloud computing node server, such as Figure 8 The computer system 800 shown. In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement of properties of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least the operations of method 170. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of the properties of a viral gene delivery vector (VGDV) sample via a separation script or computer software application that implements at least the operations of method 170. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 170. In one embodiment, the computer system is the processor of an analytical instrument that performs the measurement of viral gene delivery vector (VGDV) sample properties via a separation script or computer software application that implements at least the operations of method 170.

[0194] In one embodiment, the computer system is as follows: Figure 8 The computer system 800 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 171, 172, 173, 174, 175, and 176. In one embodiment, the computer system is as follows: Figure 8 The computer system / server 812 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 171, 172, 173, 174, 175, and 176. In one embodiment, the computer system is as follows: Figure 8 The processing unit 816 shown performs measurement of viral gene delivery vector (VGDV) samples via a separation script or computer software application that implements at least operations 171, 172, 173, 174, 175 and 176.

[0195] In one embodiment, the mass fraction X of protein in the sample is calculated. A This includes calculating the protein mass fraction X in the sample using the following formula. A :

[0196] ,

[0197] Calculate the extinction coefficient of the sample at the specified wavelength. This includes calculating the extinction coefficient of the sample at a given wavelength using the following formula. :

[0198] ,

[0199] Calculate the refractive index increment (dn / dc) of the sample. VGDV This includes calculating the refractive index increment (dn / dc) of the sample using the following formula. VGDV :

[0200] ,

[0201] Calculate the total mass m of the protein A This includes calculating the total mass m of protein using the following formula. A :

[0202] ,

[0203] And calculate the total mass m of the modifier. B The total mass m of the modifier is calculated using the following formula. B ,

[0204] .

[0205] In one embodiment, the wavelength λ is one of 260 nm and 280 nm. In one embodiment, the modifier is a nucleic acid.

[0206] Total VGDV particle concentration using UV detector and FLD

[0207] In one embodiment, a computer-implemented method, system, and computer program product includes: (1) a set of logical operations performed by a computer system to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and (2) a set of logical operations performed by the computer system relative to the area below the peak of fluorescence emission data collected from the sample at the excitation wavelength (FLD) and the ultraviolet absorbance value (A) collected from the sample at the ultraviolet wavelength (λ). λ The extinction coefficient of proteins at ultraviolet wavelength λ A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. The extinction coefficient of the modifier in the sample at ultraviolet wavelength λ A proportionality constant that relates the modifier concentration to the fluorescence intensity of FLD. To calculate the protein mass fraction X in the sample A (3) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The extinction coefficient of proteins at ultraviolet wavelengths The extinction coefficient of the modifier in the sample at ultraviolet wavelength. To calculate the extinction coefficient of the sample at ultraviolet wavelength. (4) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. and a proportionality constant relating the modifier concentration to the fluorescence intensity of FLD. To calculate the extinction coefficient of the sample at the excitation wavelength. (5) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample. VGDV (6) A set of logical operations is executed by the computer system relative to the area below the peak of the fluorescence emission data collected from the sample at the excitation wavelength (FLD) and the mass fraction of protein in the sample (X). A A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. and a proportionality constant relating the modifier concentration to the fluorescence intensity of FLD. To calculate the total mass m of the protein A The total mass m of the modifier B (7) A set of logical operations is performed by the computer system to calculate the total VGDV particle concentration C of the sample via the following formula. A :

[0208] ,

[0209] Where N is the Avogrado number, and where M A This refers to the molar mass of the capsid protein in the sample, derived from a data source. In one embodiment, at least two concentration detectors include an ultraviolet absorbance (UV) detector and a fluorescence detector (FLD) at a UV wavelength λ. In another embodiment, at least two concentration detectors are a UV detector and an FLD at a UV wavelength λ.

[0210] In one embodiment, a computer-implemented method, system, and computer program product are configured to perform: operations by a computer system executing a set of logical operations to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set of analytical instruments includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and operations by a computer system executing a set of logical operations relative to the area below the peak of fluorescence emission data collected from the sample at the excitation wavelength (FLD) and the ultraviolet absorbance value (A) collected from the sample at the ultraviolet wavelength (λ). λ The extinction coefficient of proteins at ultraviolet wavelength λ A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. The extinction coefficient of the modifier in the sample at ultraviolet wavelength λ A proportionality constant that relates the modifier concentration to the fluorescence intensity of FLD. To calculate the protein mass fraction X in the sample A A set of logical operations performed by a computer system, relative to the mass fraction X of protein in the sample. A The extinction coefficient of proteins at ultraviolet wavelengths The extinction coefficient of the modifier in the sample at ultraviolet wavelength. To calculate the extinction coefficient of the sample at ultraviolet wavelength. A set of logical operations performed by a computer system, relative to the mass fraction X of protein in the sample. A A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. and a proportionality constant relating the modifier concentration to the fluorescence intensity of FLD. To calculate the extinction coefficient of the sample at the excitation wavelength. A set of logical operations performed by a computer system, relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample. VGDV ; An operation performed by a computer system using a set of logical operations relative to the area below the peak of fluorescence emission data collected from the sample at the excitation wavelength (FLD) and the mass fraction of protein in the sample (X). A A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. and a proportionality constant relating the modifier concentration to the fluorescence intensity of FLD. To calculate the total mass m of the protein A The total mass m of the modifier B The computer system performs a set of logical operations to calculate the total VGDV particle concentration C of the sample using the following formula. A :

[0211] ,

[0212] Where N is the Avogrado number, and where M A It is the molar mass of the capsid protein in the sample, derived from a data source of capsid protein molar mass.

[0213] In one embodiment, the mass fraction X of protein in the sample is calculated. A This includes calculating the protein mass fraction X in the sample using the following formula. A :

[0214] ,

[0215] Calculate the extinction coefficient of the sample at ultraviolet wavelength. This includes calculating the extinction coefficient of the sample at ultraviolet wavelengths using the following formula. :

[0216] ,

[0217] Calculate the extinction coefficient of the sample at the excitation wavelength. This includes calculating the extinction coefficient of the sample at the excitation wavelength using the following formula. :

[0218] ,

[0219] And calculate the refractive index increment (dn / dc) of the sample. VGDV This includes calculating the refractive index increment (dn / dc) of the sample using the following formula. VGDV :

[0220] ,

[0221] Calculate the total mass m of the protein A This includes calculating the total mass m of protein using the following formula. A :

[0222] ,

[0223] And calculate the total mass m of the modifier. B The total mass m of the modifier is calculated using the following formula. B :

[0224] .

[0225] In one embodiment, the UV wavelength λ is one of 260 nm and 280 nm. In one embodiment, the modifier is a nucleic acid.

[0226] Total VGDV particle concentration using dRI and FLD

[0227] In one embodiment, a computer-implemented method, system, and computer program product includes: (1) a set of logical operations performed by a computer system to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and (2) a set of logical operations performed by the computer system relative to the area below the peak of fluorescence emission data collected from the sample at the excitation wavelength, the differential refractive index dRI of the solution containing the sample, and the refractive index coefficient (dn / dc) of the protein. A The refractive index coefficient (dn / dc) of the modifier in the sample. B A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. A proportionality constant that relates the modifier concentration to the fluorescence intensity of FLD. To calculate the mass fraction X of protein in the sample A (3) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample.VGDV (4) A set of logical operations is executed by the computer system, which are relative to the mass fraction X of protein in the sample. A A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. and a proportionality constant relating the modifier concentration to the fluorescence intensity of FLD. To calculate the extinction coefficient of the sample at the excitation wavelength. (5) A set of logical operations is performed by the computer system to calculate the total VGDV particle concentration C of the sample via the following formula. A :

[0228] ,

[0229] Where N is the Avogrado number, and m A It is the total mass of protein, and of which M A This refers to the molar mass of the capsid protein in the sample, derived from a data source. In one embodiment, at least two concentration detectors include a differential refractive index (dRI) detector and a fluorescence detector (FLD). In another embodiment, at least two concentration detectors are a dRI detector and an FLD.

[0230] In one embodiment, a computer-implemented method, system, and computer program product are configured to perform: operations by which a computer system performs a set of logical operations to analyze a viral gene delivery vector (VGDV) sample on a set of analytical instruments, including at least one separation instrument, at least one static light scattering (SLS) instrument, and at least two concentration detectors; and operations by which the computer system performs a set of logical operations relative to the area below the peak of fluorescence emission data collected from the sample at the excitation wavelength, the differential refractive index (dRI) of the solution containing the sample, and the refractive index coefficient (dn / dc) of the protein. A The refractive index coefficient (dn / dc) of the modifier in the sample. B A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. A proportionality constant that relates the modifier concentration to the fluorescence intensity of FLD. To calculate the mass fraction X of protein in the sample A A set of logical operations performed by a computer system, relative to the mass fraction X of protein in the sample. A The refractive index coefficient of proteins (dn / dc) A The refractive index coefficient (dn / dc) of the modifier in the sample. B To calculate the refractive index increment (dn / dc) of the sample. VGDVA set of logical operations performed by a computer system, relative to the mass fraction X of protein in the sample. A A proportionality constant that correlates protein concentration with fluorescence intensity relative to FLD. and a proportionality constant relating the modifier concentration to the fluorescence intensity of FLD. To calculate the extinction coefficient of the sample at the excitation wavelength. The computer system performs a set of logical operations to calculate the total VGDV particle concentration C of the sample using the following formula. A :

[0231] ,

[0232] Where N is the Avogrado number, and m A It is the total mass of protein, and of which M A It is the molar mass of the capsid protein in the sample, derived from a data source of capsid protein molar mass.

[0233] In one embodiment, the mass fraction X of protein in the sample is calculated. A This includes calculating the protein mass fraction X in the sample using the following formula. A :

[0234] ,

[0235] Calculate the refractive index increment (dn / dc) of the sample. VGDV This includes calculating the refractive index increment (dn / dc) of the sample using the following formula. VGDV :

[0236] ,

[0237] Furthermore, the extinction coefficient of the sample at the excitation wavelength was calculated. This includes calculating the extinction coefficient of the sample at the excitation wavelength using the following formula. :

[0238] . Example

[0239] For example, Figure 4A , Figure 4B and Figure 4C This method describes how to calculate particle concentration and the total VGDV particle concentration (C) of a sample. A The consistency of the first sample with the second sample is reflected in the "full:empty" ratio. Figure 4C This significantly demonstrates that the method can accurately calculate the capsid content (C) of a sample. p / V g ), (CA / C FULL Figure 4A shows representative data (chromatograms) collected by the SLS instrument at 90 degrees. Figure 4B This indicates that the method can calculate the total VGDV concentration of a sample.

[0240] Figure 5 illustrates the ability of this method to quantify the aggregate content of AAV samples. Figure 5 shows the total AAV particle concentration overlapping with the UV traces of the AAV samples. The peak eluted before the main peak (6.5 mL–7.5 mL) represents AAV aggregates. Using this method, the degree of aggregation is determined by calculating the particle concentration of all eluted data slices (chromatographic data slices).

[0241] Figure 6A Fluorescence detectors (FLDs) can also be used as one type of concentration detector. Figure 6A Representative data (chromatograms) collected by the SLS instrument, UV detector, and FLD at 90 degrees are shown. Figure 6B The method of analyzing the molar mass (MM) / molecular weight (MW) results of AAV samples using UV and FLD as concentration detectors is described, which is consistent with the expected molar mass / molecular weight results. Figure 6C The method of analyzing the molecular weight (MW) / molar mass (MM) results of AAV samples using dRI and FLD as concentration detectors is described, which is consistent with the expected molecular weight / molar mass values.

[0242] Figure 7A This shows the data collected using ion exchange chromatography (IEX) as the separation instrument. Figure 7A The data (chromatograms) collected by the SLS instrument at 90 degrees are shown. Figure 7B The data collected using a method employing field flow fractionation (FFF) as a separation instrument are shown. Figure 7B The data (chromatograms) collected by the SLS instrument at 90 degrees are shown. Figure 7C The method described is to analyze the molar mass (MM) of AAV samples by using UV and dRI as concentration detectors and FFF as separation system, which is consistent with the expected molar mass / expected molecular weight value.

[0243] Computer System

[0244] In an exemplary embodiment, the computer system is as follows: Figure 5The computer system 500 shown is merely an example of a computer system and is not intended to impose any limitation on the scope or functionality of the embodiments of the present invention. In any case, the computer system 500 can be implemented to perform and / or is capable of performing any of the functions / operations of the present invention.

[0245] Computer system 500 includes computer system / server 512, which operates in conjunction with many other general-purpose or special-purpose computing system environments or configurations. Examples of well-known computing systems, environments, and / or configurations that may be suitable for computer system / server 512 include, but are not limited to, personal computer systems, server computer systems, thin clients, fat clients, handheld or laptop devices, multiprocessor systems, microprocessor-based systems, set-top boxes, programmable consumer electronics, network PCs, minicomputer systems, mainframe computer systems, and distributed cloud computing environments that include any of the aforementioned systems or devices.

[0246] In general, a computer system / server 512 can be described as computer system executable instructions, such as program modules, executed by the computer system. Typically, program modules can include routines, programs, objects, components, logic, and / or data structures that perform specific tasks or implement specific abstract data types. The computer system / server 512 can be implemented in a distributed cloud computing environment, where tasks are performed by remote processing devices linked via a communication network. In a distributed cloud computing environment, program modules may reside on local and remote computer system storage media, including memory storage devices.

[0247] like Figure 5 As shown, the computer system / server 512 in computer system 500 is illustrated in the form of a general-purpose computing device. The components of the computer system / server 512 may include, but are not limited to, one or more processors or processing units 516, system memory 528, and a bus 518 that couples various system components, including system memory 528, to processor 516.

[0248] Bus 518 represents any one or more of several types of bus architectures, including memory buses or memory controllers, peripheral buses, accelerated graphics ports, and processor or local buses using any of the various bus architectures. As an example and not a limitation, such architectures include Industry Standard Architecture (ISA) buses, Micro Channel Architecture (MCA) buses, Enhanced ISA (EISA) buses, Video Electronics Standards Association (VESA) local buses, and Peripheral Component Interconnect (PCI) buses.

[0249] Computer system / server 512 typically includes a variety of computer system readable media. Such media can be any available media accessible to computer system / server 512, and includes both volatile and non-volatile media, as well as removable and non-removable media.

[0250] System memory 528 may include computer system readable media in the form of volatile memory, such as random access memory (RAM) 530 and / or cache memory 532. Computer system / server 512 may also include other removable / non-removable, volatile / non-volatile computer system storage media. By way of example only, storage system 534 may be provided for reading from and writing to non-removable, non-volatile magnetic media (not shown and generally referred to as "hard disk drives"). Although not shown, disk drives may be provided for reading from and writing to removable, non-volatile disks (e.g., "floppy disks"), and optical disk drives may be provided for reading from or writing to removable, non-volatile optical disks (such as CD-ROMs, DVD-ROMs, or other optical media). In this case, each may be connected to bus 518 via one or more data media interfaces. As will be further described and depicted below, memory 528 may include at least one program product having a set (e.g., at least one) of program modules configured to implement the functions / operations of embodiments of the invention.

[0251] A program / utility 540 having a set (at least one) of program modules 542 may be stored in memory 528 as an example, not as a limitation. Exemplary program module 542 may include an operating system, one or more applications, other program modules, and program data. Each of the operating system, one or more applications, other program modules, and program data, or some combination thereof, may include an implementation of a network environment. Program module 542 typically performs the functions and / or methods of embodiments of the present invention.

[0252] The computer system / server 512 can also communicate with one or more external devices 514, such as a keyboard, indicating devices, a display 524, one or more devices enabling a user to interact with the computer system / server 512, and / or any device enabling the computer system / server 512 to communicate with one or more other computing devices (e.g., a network interface card, a modem, etc.). This communication can occur through an input / output (I / O) interface 522. Furthermore, the computer system / server 512 can communicate with one or more networks, such as a local area network (LAN), a general wide area network (WAN), and / or a public network (e.g., the Internet), via a network adapter 520. As shown, the network adapter 520 communicates with other components of the computer system / server 512 via a bus 518. It should be understood that, although not shown, other hardware and / or software components can be used in conjunction with the computer system / server 512. Examples include, but are not limited to, microcode, device drivers, redundant processing units, external disk drive arrays, RAID systems, tape drives, and data archiving storage systems.

[0253] Computer program products

[0254] This invention can be a system, method, and / or computer program product. A computer program product may include a computer-readable storage medium (or medium) having computer-readable program instructions thereon for causing a processor to implement aspects of the invention.

[0255] Computer-readable storage media can be tangible devices capable of retaining and storing instructions for use by an instruction execution device. Computer-readable storage media can be, for example, but not limited to, electronic storage devices, magnetic storage devices, optical storage devices, electromagnetic storage devices, semiconductor storage devices, or any suitable combination of the foregoing. A non-exhaustive list of more specific examples of computer-readable storage media includes the following: portable computer floppy disks, hard disks, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), static random access memory (SRAM), portable optical disc read-only memory (CD-ROM), digital universal disk (DVD), memory sticks, floppy disks, mechanical encoding devices (such as punch cards or raised structures in recesses on which instructions are recorded), and any suitable combination of the foregoing. As used herein, computer-readable storage media should not be construed as transient signals themselves, such as radio waves or other freely propagating electromagnetic waves, electromagnetic waves propagating through waveguides or other transmission media (e.g., light pulses through fiber optic cables), or electrical signals transmitted through wires.

[0256] The computer-readable program instructions described herein can be downloaded from computer-readable storage media to various computing / processing devices, or downloaded via networks such as the Internet, local area networks, wide area networks, and / or wireless networks to external computers or external storage devices. Networks may include copper transmission cables, optical fiber transmission, wireless transmission, routers, firewalls, switches, gateway computers, and / or edge servers. A network adapter card or network interface in each computing / processing device receives the computer-readable program instructions from the network and forwards them to a computer-readable storage medium within the corresponding computing / processing device.

[0257] Computer-readable program instructions used to implement the operations of this invention may be assembly instructions, instruction set architecture (ISA) instructions, machine instructions, machine-dependent instructions, microcode, firmware instructions, state setting data, or source code or object code written in any combination of one or more programming languages, including object-oriented programming languages ​​such as Smalltalk, C++, or the like, and traditional procedural programming languages ​​such as the "C" programming language or similar programming languages. The computer-readable program instructions may execute entirely on the user's computer, partially on the user's computer as a standalone software package, partially on the user's computer and partially on a remote computer, or entirely on a remote computer or server. In the latter case, the remote computer may be connected to the user's computer via any type of network, including a local area network (LAN) or a wide area network (WAN), or may be connected to an external computer (e.g., via the Internet using an Internet service provider). In some embodiments, electronic circuitry including, for example, programmable logic circuitry, a field-programmable gate array (FPGA), or a programmable logic array (PLA) may execute the computer-readable program instructions to personalize the electronic circuitry in order to perform aspects of this invention by utilizing state information from the computer-readable program instructions.

[0258] This document describes aspects of the invention with reference to flowchart illustrations and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of the invention. It should be understood that each block in the flowchart illustrations and / or block diagrams, and combinations of blocks in the flowchart illustrations and / or block diagrams, can be implemented by computer-readable program instructions.

[0259] These computer-readable program instructions can be provided to a processor of a general-purpose computer, a special-purpose computer, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, create means for implementing the functions / actions specified in one or more blocks of the flowchart and / or block diagram. These computer-readable program instructions can also be stored in a computer-readable storage medium that can instruct a computer, programmable data processing apparatus, and / or other device to operate in a particular manner, such that the computer-readable storage medium storing the instructions includes an article of manufacture comprising instructions that implement aspects of the functions / actions specified in one or more blocks of the flowchart and / or block diagram.

[0260] Computer-readable program instructions may also be loaded onto a computer, other programmable data processing apparatus or other device to cause a series of operational steps to be performed on the computer, other programmable apparatus or other device to produce a computer-implemented process, such that the instructions, which execute on the computer, other programmable apparatus or other device, perform the function / action specified in one or more boxes of a flowchart and / or block diagram.

[0261] The flowcharts and block diagrams in the accompanying drawings illustrate the architecture, functionality, and operation of possible implementations of systems, methods, and computer program products according to various embodiments of the present invention. In this regard, each box in a flowchart or block diagram may represent a portion of a module, segment, or instruction, which includes one or more executable instructions for implementing one or more specified logical functions. In some alternative implementations, the functions indicated in the boxes may occur outside the order shown in the figures. For example, two boxes shown consecutively may actually be executed substantially simultaneously, or these boxes may sometimes be executed in reverse order, depending on the functions involved. It should also be noted that each box in the block diagrams and / or flowcharts, and combinations of boxes in the block diagrams and / or flowcharts, may be implemented by a dedicated hardware-based system that performs the specified function or action or implements a combination of dedicated hardware and computer instructions.

[0262] The various embodiments described herein have been presented for illustrative purposes and are not intended to be exhaustive or limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein has been chosen to explain the principles of the embodiments, their practical application, or technical improvements to technologies found in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A computer-implemented method for measuring properties of a viral gene delivery vector (VGDV) sample, comprising: A set of logical operations executed by a computer system to analyze a sample of the viral gene delivery vector VGDV on a set of analytical instruments. The amount of capsid protein m of the sample was obtained. A The mass m of the modifier in the sample B The molar mass M of the modifier in the sample B and the molar mass M of the capsid protein of the sample A The molar mass M of the capsid protein of the sample was determined. A The data is stored in a capsid protein molar mass data source, wherein the group includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors; The molar mass M of the capsid protein of the sample is received from the capsid protein molar mass data source. A ; Receive the injection volume v of the sample from the injection volume data source; and The computer system performs the calculation of the total VGDV particle concentration C of the sample using the following formula. A A set of logical operations: ， Where N is the Avogrado number.

2. The method of claim 1, wherein the analysis comprises analyzing the sample on the group via analytical techniques. The analytical technique described therein is one of viral vector analysis, protein conjugate analysis, and copolymer composition analysis.

3. The method according to claim 1, wherein the at least one separation instrument comprises at least one of a size exclusion chromatography unit, a field flow fractionation unit, and an ion exchange chromatography unit.

4. The method according to claim 1, wherein the at least one static light scattering instrument comprises a multi-angle light scattering instrument.

5. The method according to claim 1, wherein the at least two concentration detectors comprise a first ultraviolet absorbance detector at a first wavelength λ1 and a second ultraviolet absorbance detector at a second wavelength λ2.

6. The method according to claim 5, Wherein the first wavelength λ1 is 260nm, and The second wavelength λ2 is 280 nm.

7. The method of claim 1, wherein the at least two concentration detectors comprise an ultraviolet absorbance detector at wavelength λ and a differential refractive index detector.

8. The method according to claim 7, wherein the wavelength λ is one of 260 nm and 280 nm.

9. The method of claim 1, wherein the at least two concentration detectors comprise an ultraviolet absorbance detector and a fluorescence detector at wavelength λ.

10. The method of claim 9, wherein the wavelength λ is one of 260 nm and 280 nm.

11. The method of claim 1, wherein the at least two concentration detectors comprise a differential refractive index detector and a fluorescence detector.

12. The method according to claim 1, wherein the mass m of the modifier in the sample B This refers to the nucleic acid quality of the sample.

13. The method according to claim 1, wherein the molar mass M of the modifier in the sample is... B It is the molar mass of the nucleic acid in the sample.

14. The method according to claim 1, further comprising: The computer system receives the molar mass M of the total modifier in the entire VGDV sample from the total modifier molar mass data source. Full ; The computer system performs the calculation of the total VGDV concentration C of the total VGDV sample using the following formula. Full A set of logical operations: ;and The computer system performs the calculation of the empty VGDV concentration C of the full VGDV sample using the following formula. Empty A set of logical operations: 。 15. The method of claim 14, further comprising: The computer system executes a set of logical operations to analyze the full VGDV sample on the group. The molar mass M of the total modifier in the total VGDV sample was obtained. Full ;and The molar mass M of the fully modified agent in the full VGDV sample. Full It is stored in the total modifier molar mass data source.

16. The method of claim 14, wherein the modifier is a nucleic acid.

17. The method according to claim 1, further comprising: The computer system executes a set of logical operations to analyze the entire VGDV signal region of the sample on the group and to analyze the clustered peak region of the sample on the group. The total peak protein mass m of the VGDV of the sample corresponding to the entire VGDV signal region of the sample was obtained. A,ent The mass m of the VGDV peak modifier of the sample corresponding to the entire VGDV signal region of the sample. B,ent The molar mass M of the entire peak protein of the VGDV in the sample corresponding to the entire VGDV signal region of the sample. A,ent The molar mass M of the modifier for the entire VGDV peak of the sample corresponding to the entire VGDV signal region of the sample. B,ent The mass m of the VGDV aggregation peak protein of the sample corresponding to the aggregation peak region of the sample. A,agg The mass m of the VGDV aggregation peak modifier of the sample corresponding to the aggregation peak region of the sample. B,agg The molar mass M of the VGDV aggregation peak protein in the sample corresponding to the aggregation peak region of the sample. A,agg The molar mass M of the VGDV aggregation peak modifier of the sample corresponding to the aggregation peak region of the sample. B,agg ; and The computer system performs the following calculation to determine the total VGDV peak particle concentration C of the sample corresponding to the entire VGDV signal region of the sample. A, ent A set of logical operations: ； The computer system performs the following calculation to determine the total VGDV peak particle concentration C of the sample corresponding to the peak aggregation region of the sample. A,agg A set of logical operations: 。 18. The method of claim 17, further comprising: The computer system executes a set of logical operations to analyze the entire VGDV signal region of the whole viral gene delivery vector sample on the group and to analyze the aggregation peak region of the whole sample on the group. The peak molar mass M of the VGDV of the entire modifier within the entire sample, corresponding to the entire VGDV signal region of the entire sample, is obtained. Full,ent And the VGDV aggregation peak molar mass M of the entire modifier within the entire sample corresponding to the aggregation peak region of the entire sample. Full,agg ; The computer system performs the following calculation to determine the total peak VGDV concentration C of the whole sample corresponding to the entire VGDV signal region of the VGDV sample: Full,ent A set of logical operations: ； The computer system performs the following calculation to determine the total VGDV concentration C of the whole sample corresponding to the aggregation peak region of the whole sample via the following formula: Full,agg A set of logical operations: ； The computer system performs the following calculation to determine the peak empty VGDV concentration C of the whole sample corresponding to the entire VGDV signal region of the whole sample. Empty,ent A set of logical operations: ； The computer system performs the following calculation to determine the VGDV aggregation peak empty VGDV concentration C of the whole sample corresponding to the aggregation peak region of the whole sample. Empty,agg A set of logical operations: 。 19. A computer-implemented method for measuring properties of a viral gene delivery vector (VGDV) sample, comprising: A computer system executes a set of logical operations to analyze a sample of the viral gene delivery vector VGDV on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering instrument, and at least one concentration detector. The mass m of the modifier for the sample was obtained. B The molar mass M of the modifier in the sample B and at least one UV extinction coefficient of the sample; The computer system performs calculations of the capsid protein mass m of the sample relative to at least one refractive index increment value from the at least one concentration detector. A and the molar mass M of the capsid protein of the sample A A set of logical operations; Receive the injection volume v of the sample from the injection volume data source; as well as The computer system performs a set of logical operations to calculate the total VGDV particle concentration CA of the sample via the following formula: Where N is the Avogrado number.

20. A computer-implemented method for measuring properties of a viral gene delivery vector (VGDV) sample, comprising: A set of logical operations performed by a computer system to analyze a sample of the viral gene delivery vector VGDV on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors; The computer system performs the calculation of the protein mass fraction X in the sample relative to the following parameters. A A set of logical operations: the ultraviolet absorbance value A collected from the sample at the first wavelength λ1. λ1 The ultraviolet absorbance value A collected from the sample at the second wavelength λ2 λ2 The extinction coefficient of the protein at the first wavelength λ1 The extinction coefficient of the protein at the second wavelength λ2 The extinction coefficient of the modifier in the sample at the first wavelength λ1 and the extinction coefficient of the modifier in the sample at the second wavelength λ2 ; The computer system performs the calculation of the extinction coefficient of the sample at the first wavelength relative to the following parameters. A set of logical operations: the mass fraction of the protein in the sample X A The extinction coefficient of the protein at the first wavelength and the extinction coefficient of the modifier in the sample at the first wavelength. ; The extinction coefficient of the sample at the second wavelength is calculated by a computer system relative to the following parameters. A set of logical operations: the mass fraction X of the protein in the sample A The extinction coefficient of the protein at the second wavelength and the extinction coefficient of the modifier in the sample at the second wavelength. ; The computer system performs the calculation of the refractive index increment (dn / dc) of the sample relative to the following parameters. VGDV A set of logical operations: the mass fraction of the protein in the sample X A The refractive index coefficient (dn / dc) of the protein. A The refractive index coefficient (dn / dc) of the modifier in the sample. B ; The computer system performs the calculation of the total mass m of the protein relative to the following parameters. A The total mass m of the modifier B A set of logical operations: the ultraviolet absorbance value A collected from the sample at wavelength λ. λ The mass fraction X of the protein in the sample A The extinction coefficient of the protein at the wavelength λ The extinction coefficient of the modifier in the sample at the wavelength λ The wavelength λ is one of the first wavelength λ1 and the second wavelength λ2; and The computer system performs the calculation of the total VGDV particle concentration C of the sample using the following formula. A A set of logical operations: ， Where N is the Avogrado number. Where M A It is the molar mass of the capsid protein in the sample, derived from a data source of capsid protein molar mass.

21. The method according to claim 20, The mass fraction X of the protein in the sample is calculated therein. A This includes calculating the mass fraction X of the protein in the sample using the following formula. A : ， The calculation of the extinction coefficient of the sample at the first wavelength is described above. This includes calculating the extinction coefficient of the sample at the first wavelength using the following formula. : ， The calculation of the extinction coefficient of the sample at the second wavelength is described above. This includes calculating the extinction coefficient of the sample at the second wavelength using the following formula. : ， The calculation of the refractive index increment (dn / dc) of the sample is described above. VGDV This includes calculating the refractive index increment (dn / dc) of the sample using the following formula. VGDV : ， The total mass m of the protein is calculated as described above. A This includes calculating the total mass m of the protein using the following formula. A : ， The total mass m of the modifier is calculated as follows. B This includes calculating the total mass m of the modifier using the following formula. B : 。 22. The method according to claim 20, Wherein the first wavelength λ1 is 260 nm, and The second wavelength λ2 is 280 nm.

23. The method of claim 20, wherein the modifier is a nucleic acid.

24. A computer-implemented method for measuring properties of a viral gene delivery vector (VGDV) sample, comprising: A set of logical operations performed by a computer system to analyze a sample of the viral gene delivery vector VGDV on a set of analytical instruments, wherein the set includes at least one separation instrument, at least one static light scattering instrument, and at least two concentration detectors; The computer system performs the calculation of the protein mass fraction X in the sample relative to the following parameters. A A set of logical operations: the ultraviolet absorbance value A collected from the sample at wavelength λ. λ The refractive index coefficient (dn / dc) of the modifier in the sample. B The differential refractive index dRI of the solution containing the sample, and the extinction coefficient of the modifier at the wavelength λ. The extinction coefficient of the protein at the wavelength λ The refractive index coefficient (dn / dc) of the protein. A ; The computer system performs the calculation of the extinction coefficient of the sample at the wavelength relative to the following parameters. A set of logical operations: the mass fraction of the protein in the sample X A The extinction coefficient of the protein at the wavelength. and the extinction coefficient of the modifier in the sample at the wavelength. ; The computer system performs the calculation of the refractive index increment (dn / dc) of the sample relative to the following parameters. VGDV A set of logical operations: the mass fraction of the protein in the sample X A The refractive index coefficient (dn / dc) of the protein. A The refractive index coefficient (dn / dc) of the modifier in the sample. B ; The computer system performs the calculation of the total mass m of the protein relative to the following parameters. A The total mass m of the modifier B A set of logical operations: the differential refractive index dRI of the solution containing the sample, the mass fraction X of the protein in the sample. A The refractive index coefficient (dn / dc) of the protein. A and the refractive index coefficient (dn / dc) of the modifier in the sample. B ;as well as The computer system performs the calculation of the total VGDV particle concentration C of the sample using the following formula. A A set of logical operations: ， Where N is the Avogrado number. Where M A It is the molar mass of the capsid protein in the sample, derived from a data source of capsid protein molar mass.

25. The method according to claim 24, The mass fraction X of the protein in the sample is calculated therein. A This includes calculating the mass fraction X of the protein in the sample using the following formula. A : The calculation of the extinction coefficient of the sample at the wavelength is described above. This includes calculating the extinction coefficient of the sample at the wavelength using the following formula. : ， The calculation of the refractive index increment (dn / dc) of the sample is described above. VGDV This includes calculating the refractive index increment (dn / dc) of the sample using the following formula. VGDV : ， The total mass m of the protein is calculated as described above. A This includes calculating the total mass m of the protein using the following formula. A : ， The total mass m of the modifier is calculated as follows. B This includes calculating the total mass m of the modifier using the following formula. B : 。 26. The method of claim 24, wherein the wavelength λ is one of 260 nm and 280 nm.

27. The method of claim 24, wherein the modifier is a nucleic acid.

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