Control of Polymer Solution Purification via Real-Time Multi-Angle Light Scattering

Abstract

The present disclosure describes a computer-implemented method, system, and computer program product for controlling the purification of a polymer solution via real-time multi-angle light scattering.

Description

Technical Field

[0001] (Related Art) This application claims the benefit of the filing date of U.S. Patent Application No. 17 / 855,791, filed on June 30, 2022, entitled "CONTROLLING THE PURIFICATION OF A MACROMOLECULE SOLUTION VIA REAL-TIME MULTI-ANGLE LIGHT SCATTERING", which is hereby incorporated by reference in its entirety.

Background Art

[0002] The present disclosure relates to multi-angle light scattering, and more specifically, to controlling the purification of a macromolecule solution by real-time multi-angle light scattering.

Summary of the Invention

[0003] The present disclosure describes a computer-implemented method, system, and computer program product for controlling the purification of a macromolecule solution via real-time multi-angle light scattering. In an exemplary embodiment, the computer-implemented method, system, and computer program product (1) receive, by a computer system, from a set of devices, a baseline multi-angle light scattering (MALS) signal value and a baseline ultraviolet (UV) signal value of a pure buffer flowing from a chromatographic purification system to the set of devices, the set of devices including a MALS device and a UV detector, and (2) in response to receiving the baseline MALS signal value from the MALS device, (a) receive, by the computer system, from the MALS device, scattering intensity values of a sample solution flowing from the chromatographic purification system to the MALS device over time, the sample solution including at least one type of macromolecule, (b) calculate, by the computer system, an average of the baseline MALS signal values, and (c) subtract, by the computer system, the average of the baseline MALS signal values from the scattering intensity values to obtain an excess scattering intensity value Iscatt of the intensity time seriesj obtaining; and in response to receiving a baseline UV signal value from a (3) UV detector, (a) the computer system receives, from the UV detector, the UV absorption values of the sample solution flowing from the chromatographic purification system to the UV detector over time, (b) the computer system calculates the average of the baseline UV signal values, (c) the computer system executes a set of logical operations to apply UV absorption alignment correction for UV absorption alignment parameters and UV absorption band broadening correction for UV absorption band broadening parameters to the UV absorption values over time for the UV detector and the MALS instrument to obtain corrected UV absorption values over time, (d) the computer system subtracts the average of the baseline UV signal values from the corrected UV absorption values to obtain the excess UV absorption values of the UV time series; and (4) the computer system determines the concentration value c of the sample solution with respect to the excess UV absorption values of the UV time series j and the absorption coefficient of the sample solution according to Beer's law; and (5) the computer system receives the pH value of the sample solution over time from a pH detector and the conductivity value of the sample solution over time from a conductivity detector, the set of instruments further including a pH detector and a conductivity detector; and (6) the computer system executes a set of logical operations to (a) apply pH alignment correction for pH alignment parameters to the pH values over time for the pH detector and the MALS instrument to obtain corrected pH values over time, and (b) apply conductivity alignment correction for conductivity alignment parameters to the conductivity values over time for the conductivity detector and the MALS instrument to obtain corrected conductivity values over time, to obtain a plurality of signals including the excess scattering intensity value Iscatt of the intensity time series j the concentration value c of the sample solution j the corrected pH value, and the corrected conductivity value; and (7) the computer system determines time ranges t1, t2,... t nPerforming a set of logical operations that average each of a plurality of signals, wherein the time series includes a time range, and the time range corresponds to a fraction of a sample solution collected by a fraction collector connected to a set of devices; performing to obtain average values [lscatt, c, pH, cond]1, [lscatt, c, pH, cond]2, ..., [lscatt, c, pH, cond] n Obtaining (8) aggregate content values %agg1, %agg2, ... %agg of fractions of the sample solution measured by the SEC-MALS instrument by a computer system n , and weight average molar mass values M1, M2, ... M n Receiving (9) by a computer system, the aggregate content values %agg1, %agg2, ... %agg n , and weight average molar mass values M1, M2, ... M n Storing, by the computer system, the aggregate content values %agg1, %agg2, ... %agg n , and weight average molar mass values M1, M2, ... M with respect to the average values [lscatt, c, pH, cond]1, [lscatt, c, pH, cond]2, ... [lscatt, c, pH, cond] in a macromolecular characterization look-up table (MCLUT).

Brief Description of the Drawings

[0004]

Figure 1A

Figure 1B

Figure 1C

Figure 2A

Figure 2B

Figure 2C

Figure 3

Figure 4

Figure 5

Best Mode for Carrying Out the Invention

[0005] The present disclosure describes a computer-implemented method, system, and computer program product for controlling the purification of a polymer solution via real-time multi-angle light scattering. In an exemplary embodiment, the computer-implemented method, system, and computer program product include: (1) receiving, by a computer system, from a set of devices, a baseline multi-angle light scattering (MALS) signal value and a baseline ultraviolet (UV) signal value of a pure buffer flowing from a chromatographic purification system to the set of devices, the set of devices including a MALS device and a UV detector; and (2) in response to receiving the baseline MALS signal value from the MALS device, (a) receiving, by the computer system, from the MALS device, scattering intensity values of a sample solution flowing from the chromatographic purification system to the MALS device over time, the sample solution including at least one type of polymer, (b) calculating, by the computer system, an average of the baseline MALS signal values, and (c) subtracting, by the computer system, the average of the baseline MALS signal values from the scattering intensity values to obtain an excess scattering intensity value Iscatt of the intensity time series jobtaining; in response to receiving a baseline UV signal value from a (3) UV detector, (a) a computer system receives, from the UV detector, the UV absorption values of the sample solution flowing from the chromatographic purification system to the UV detector over time, (b) the computer system calculates the average of the baseline UV signal values, (c) the computer system executes a set of logical operations for applying UV absorption alignment correction for UV absorption alignment parameters and UV absorption band broadening correction for UV absorption band broadening parameters to the UV absorption values over time for the UV detector and the MALS instrument to obtain corrected UV absorption values over time, (d) the computer system subtracts the average of the baseline UV signal values from the corrected UV absorption values to obtain the excess UV absorption values of the UV time series; (4) the computer system calculates the concentration value c of the sample solution j and the absorption coefficient of the sample solution according to Beer's law; (5) the computer system receives the pH value of the sample solution over time from a pH detector and the conductivity value of the sample solution over time from a conductivity detector, the set of instruments further including a pH detector and a conductivity detector; (6) the computer system executes a set of logical operations for (a) applying pH alignment correction for pH alignment parameters to the pH values over time for the pH detector and the MALS instrument to obtain corrected pH values over time, and (b) applying conductivity alignment correction for conductivity alignment parameters to the conductivity values over time for the conductivity detector and the MALS instrument to obtain corrected conductivity values over time, to obtain a plurality of signals including the excess scattering intensity value Iscatt of the intensity time series j of the sample solution, the concentration value c j of the sample solution, the corrected pH value, and the corrected conductivity value; (7) the computer system determines time ranges t1, t2,... t nexecuting a set of logical operations that average each of a plurality of signals over a time series that includes a time range corresponding to fractions of a sample solution collected by a fraction collector connected to a set of devices, to obtain average values [lscatt,c,pH,cond]1, [lscatt,c,pH,cond]2, ... [lscatt,c,pH,cond] n obtaining, by a computer system, aggregate content values %agg1, %agg2, ... %agg of fractions of a sample solution measured by an SEC-MALS instrument n and weight average molar mass values, M1, M2, ... M n receiving, by a computer system, the aggregate content values %agg1, %agg2, ... %agg n and the weight average molar mass values M1, M2, ... M n storing, by the computer system, the aggregate content values %agg1, %agg2, ... %agg n and the weight average molar mass values M1, M2, ... M with respect to the average values [lscatt,c,pH,cond]1, [lscatt,c,pH,cond]2, ... [lscatt,c,pH,cond] in a polymer characterization look-up table (MCLUT). In one embodiment, the absorption coefficient of the sample solution is a characteristic of the sample solution. In one embodiment, the absorption coefficient is received from a user input.

[0006] In one embodiment, the method, system, and computer program product improve the chromatographic purification of monoclonal antibodies and other protein-based biopharmaceuticals by real-time MALS by overcoming obstacles to identifying the elution of aggregate proteins that must be separated from monomeric proteins. In one embodiment, the method, system, and computer program product divert the process stream to "pool" when the aggregate level is low enough and to "waste" otherwise.

[0007] In one embodiment, a method, a system, and a computer program product utilize RT-MALS and additional real-time measurements to address the need to determine online and in real-time whether aggregates are present in the eluate in order to trigger optimal monomer harvesting in a pool. In one embodiment, the method, the system, and the computer program product address the complexities and interfering factors resulting from changes in protein concentration and buffer conditions.

[0008] Definition Particle Particles can be components of a liquid sample aliquot. Such particles can be various types and sizes of molecules, nanoparticles, virus-like particles, liposomes, emulsions, bacteria, and colloids. These particles can range in size from nanometers to microns.

[0009] Analysis of macromolecular or particle species in solution Analysis of macromolecular or particle species in solution can be achieved by preparing a sample in a suitable solvent and then injecting an aliquot thereof into a separation system such as a liquid chromatography (LC) column or a field flow fractionation (FFF) channel where different species of particles contained in the sample are separated into their various components. Once separated, generally based on size, mass, or column affinity, the sample can be subjected to analysis by light scattering, refractive index, ultraviolet absorption, electrophoretic mobility, and viscosity response.

[0010] Light scattering Light scattering (LS) is a non-invasive technique for characterizing macromolecules and a wide range of particles in solution. Two types of light scattering detection frequently used for macromolecule characterization are static light scattering and dynamic light scattering.

[0011] Dynamic light scattering Dynamic light scattering is also known as quasi-elastic light scattering (QELS) and photon correlation spectroscopy (PCS). In a DLS experiment, a high-speed photodetector is used to measure the time-dependent fluctuations of the scattered light signal. DLS measurements determine the diffusion coefficient of molecules or particles, which can then be used to calculate their hydrodynamic radius.

[0012] Static light scattering Static light scattering (SLS) includes various techniques such as single angle light scattering (SALS), dual angle light scattering (DALS), low angle light scattering (LALS), and multi-angle light scattering (MALS). SLS experiments generally involve measuring the absolute intensity of light scattered from a sample in a solution irradiated by a narrow beam of light. Such measurements are often used to determine the size and structure of sample molecules or particles for appropriate classes of particles / molecules and, when combined with knowledge of the sample concentration, to determine the weight-average molar mass. In addition, the non-linearity of the intensity of scattered light as a function of sample concentration can be used to measure inter-particle interactions and associations.

[0013] Multi-angle light scattering Multi - angle light scattering (MALS) is an SLS technique for measuring light scattered by a sample at multiple angles. This is used to determine both the absolute molar mass and the average size of molecules in solution by detecting how the molecules scatter light. Parallel light from a laser light source is most frequently used, in which case this technique can be referred to as multiangle laser light scattering (MALLS). The term “multi - angle” refers to the detection of scattered light at different discrete angles, measured, for example, by a single detector that is moved over a range that includes a particular selected angle, or an array of detectors fixed at particular angular positions.

[0014] MALS measurements require a set of auxiliary elements. Among the most important of these is a collimated or focused light beam (usually from a laser source that produces a collimated beam of monochromatic light) that irradiates the region of the sample. The beam is generally plane - polarized perpendicular to the measurement plane, although other polarizations can be used, particularly when examining anisotropic particles. Another necessary element is an optical cell for holding the sample being measured. Alternatively, a cell incorporating means to enable the measurement of a flowing sample can be used. When attempting to measure the scattering properties of single particles, means must be provided to introduce such particles one by one through the light beam at a point approximately equidistant from the surrounding detectors.

[0015] Most MALS - based measurements are carried out in a plane containing a set of detectors that are usually equidistant from the sample located at the center through which the irradiation beam passes, although three - dimensional versions have also been developed, in which case the detectors are on the surface of a sphere and the sample is controlled to pass through its center, intersecting the path of the incident light beam that passes along the diameter of the sphere. The MALS technique generally sequentially collects multiplexed data from the outputs of a set of discrete detectors. A MALS light scattering photometer generally has multiple detectors.

[0016] The different detectors within a MALS detector may (i) have slightly different quantum efficiencies and different gains, and (ii) view different geometric scattering volumes, so it may be necessary to normalize the signals captured by the photodetectors of the MALS detector at each angle. If these differences are not normalized, the results of the MALS detector will be meaningless and may be inappropriately weighted for different detector angles.

[0017] Concentration detector Differential refractive index detector A differential refractive index detector (differential refractive index, dRI), or differential refractometer, or refractive index detector (differential refractometer, RI or refractive index detector, RID) is a detector that measures the refractive index of an analyte relative to a solvent. They are often used as detectors for high performance liquid chromatography and size exclusion chromatography. dRI can detect those with a refractive index different from that of the solvent, but is considered a general-purpose detector because of its low sensitivity. When light exits one material and enters another, the light bends or refracts. The refractive index of a material is a measure of how much the light bends when it is incident.

[0018] The differential refractive index detector includes a flow cell having two parts, one for the sample and one for the reference solvent. The dRI measures the refractive index of both components. When only the solvent passes through the sample component, the measured refractive indices of both components are the same, but when the analyte passes through the flow cell, the two measured refractive indices are different. This difference appears as a peak in the chromatogram. The differential refractive index detector is often used for the analysis of polymer samples in size exclusion chromatography. The dRI can output a concentration detector signal value corresponding to the concentration value of the sample.

[0019] Ultraviolet-visible spectroscopy Ultraviolet-visible spectroscopy or ultraviolet-visible spectrophotometry (UV-Vis or UV / Vis) refers to absorption spectroscopy or reflectance spectroscopy in the ultraviolet-visible spectral region. Ultraviolet-visible detectors / ultraviolet-visible spectrophotometers use light in the visible and adjacent ranges, and the absorption or reflectance within the visible range directly affects the perceived color of the chemical substances involved. In this region of the electromagnetic spectrum, atoms and molecules undergo electronic transitions. Such absorption spectroscopy measures the transition from the ground state to the excited state. The ultraviolet-visible detector / ultraviolet-visible spectrophotometer measures the intensity of the light passing through the sample (I) and compares it with the intensity of the light before passing through the sample (I o ), where the ratio I / I o is called the transmittance and is usually expressed as a percentage (%T). The absorbance A is based on the transmittance according to the following equation. A = -log(%T / 100%).

[0020] The UV-visible spectrophotometer can also be configured to measure reflectance. The spectrophotometer measures the intensity of the light reflected from the sample (I) and compares it with the intensity of the light reflected from the reference material (I o ), and the ratio I / I o is called the reflectance and is usually expressed as a percentage (%R). The ultraviolet absorption detector can output a concentration detector signal value corresponding to the concentration value of the sample.

[0021] Current technology In the process of generating therapeutic proteins such as monoclonal antibodies (mAbs), the proteins are produced by the fermentation of cells within a bioreactor. Substantial purification from lysates, nucleic acids, and most host cell proteins is readily achieved, but the pure monomeric form of the protein needs to be further purified from unwanted aggregates. Often, such purification is accomplished by chromatographic means such as size exclusion chromatography, ion exchange chromatography, hydrophobic interaction chromatography, affinity chromatography, or membrane chromatography. In the process of chromatographic purification, the desired and undesired components are dissolved or suspended in solution and flow through a stationary phase such as a column or membrane, or are first loaded onto the column or membrane under one buffer condition (loading buffer) and then eluted from the column or membrane under one or more buffer conditions different from the loading buffer (elution buffer).

[0022] As a result of flowing through the stationary phase, various components such as monomers and aggregates are separated and elute at different times. However, in a typical production process, some portion of the aggregates elutes with the pure monomer at a ratio that varies with elution conditions such as time and / or buffer salt concentration. Specific fractions of the eluted material are collected and pooled under the assumption that they contain the desired components, in this case the pure monomer, and that the mixing of unwanted components is minimal. Optimizing the collection of the monomer and maintaining an accurate cut-off point to reduce the aggregate components within an acceptable level are important issues that affect yield and profitability.

[0023] To characterize the purification process, multiple test fractions are obtained and analyzed on separate off-line analytical instruments to determine the type and amount of solution components in each. These characteristics correlate with process parameters such as elution time and signal levels from concentration detectors such as UV absorption detectors. During the actual purification operation, pools of material are then collected according to the times and signal levels corresponding to those that produced optimal results with respect to purity and amount for the test fractions. The final pool may be considered sufficiently pure or may be processed through additional purification steps to remove further impurities. However, none of the information obtained online during the purification process is sufficiently definitive to determine whether aggregates are actually present in the pool or their amounts. Additional off-line tests must be performed to confirm that the correct fractions are being pooled. Unintentional variations in elution conditions, column aging, and other factors can change the monomer-aggregate co-elution characteristics relative to the initial characterization of the test fractions, leading to either unacceptable aggregate levels in the pool (requiring either rejection and repurification or lot discard) or loss of valuable monomers that should have been collected.

[0024] A variety of methods that meet this need, including dual-wavelength UV absorbance, Raman scattering, and static light scattering (the latter including MALS), have been proposed and tested. Most of these measurement techniques have generally not been found to be sufficiently sensitive to the presence of aggregates to meet industrial requirements. MALS provides a signal related to the product of the weight-average molar mass M w and the concentration c of the protein in solution, and in principle has sufficient sensitivity since the presence of aggregates causes M w to increase disproportionately and thus the light scattering amplitude to increase. However, under typical purification conditions (high protein concentrations that vary during elution and / or changing pH and salt concentrations), the influence of these parameters on the light scattering amplitude can be greater than the desired sensitivity to aggregates, reducing the suitability of MALS for application.

[0025] Therefore, in order to identify the optimal fractions to be collected upon elution with critical knowledge, it is advantageous to implement an apparatus and method for determining online and in real time whether aggregates are present in the eluate and their concentrations. Therefore, there is a need to control the purification of polymer solutions via real-time multi-angle light scattering.

[0026] In one embodiment, FIGS. 1A, 1B, and 1C depict the hardware / equipment configuration of a method, system, and computer product. Also, in one embodiment, FIG. 3 depicts times t1, t2, t3, t4, and t5 of a method, system, and computer product. In one embodiment, FIG. 4 depicts times t1, t2, t3, t4, and t5 of a method, system, and computer product.

[0027] Referring to FIG. 2, in an exemplary embodiment, a computer-implemented method, system, and computer program product include receiving, by a computer system, from a set of devices, a baseline multi-angle light scattering (MALS) signal value and a baseline ultraviolet (UV) signal value of a pure buffer flowing from a chromatography purification system to the set of devices, the set of devices including a MALS device and a UV detector, operation 110 of receiving; in response to receiving the baseline MALS signal value from the MALS device, (a) receiving, by the computer system, from the MALS device, scattering intensity values of a sample solution flowing from the chromatography purification system to the MALS device over time, the sample solution including at least one type of polymer, (b) calculating, by the computer system, an average of the baseline MALS signal values, and (c) subtracting, by the computer system, the average of the baseline MALS signal values from the scattering intensity values to obtain an excess scattering intensity value Iscatt of the intensity time series jThe operation 112 of obtaining, and in response to receiving the baseline UV signal value from the UV detector, (a) the computer system receives from the UV detector the UV absorption values of the sample solution flowing from the chromatographic purification system to the UV detector over time, (b) the computer system calculates the average of the baseline UV signal values, (c) the computer system executes a set of logical operations that apply UV absorption alignment correction for UV absorption alignment parameters and UV absorption band expansion correction for UV absorption band expansion parameters to the time-series UV absorption values for the UV detector and the MALS instrument to obtain corrected UV absorption values over time, (d) the operation 114 of the computer system subtracting the average of the baseline UV signal values from the corrected UV absorption values to obtain the excess UV absorption values of the UV time series, and the computer system calculates the concentration value c of the sample solution with respect to the excess UV absorption values of the UV time series j and the absorption coefficient of the sample solution according to Beer's law in the operation 116, and the computer system receives the pH values of the sample solution over time from the pH detector and the conductivity values of the sample solution over time from the conductivity detector, wherein the set of instruments further includes the pH detector and the conductivity detector, in the receiving operation 118, and the computer system (a) applies pH alignment correction for pH alignment parameters to the pH values over time for the pH detector and the MALS instrument to obtain corrected pH values over time, and (b) applies conductivity alignment correction for conductivity alignment parameters to the conductivity values over time for the conductivity detector and the MALS instrument to obtain corrected conductivity values over time, executes a set of logical operations to obtain the excess scattering intensity value Iscatt of the intensity time series j of the sample solution, the concentration value c j of the sample solution, the corrected pH value, and a plurality of signals including the corrected conductivity value in the operation 120, and the computer system determines the time ranges t1, t2,... t nexecuting a set of logical operations that average each of a plurality of signals over a time series that includes a time range corresponding to fractions of a sample solution collected by a fraction collector connected to a set of devices, to obtain average values [lscatt,c,pH,cond]1, [lscatt,c,pH,cond]2,... [lscatt,c,pH,cond] n an operation 122 to obtain n , and a computer system receives aggregate content values %agg1, %agg2,... %agg of fractions of a sample solution measured by an SEC-MALS device n , and weight average molar mass values, M1, M2,... M n in an operation 124, and the computer system stores the aggregate content values %agg1, %agg2,... %agg n , and the weight average molar mass values M1, M2,... M n with respect to the average values [lscatt,c,pH,cond]1, [lscatt,c,pH,cond]2,... [lscatt,c,pH,cond] n in a polymer characterization look-up table (MCLUT) in an operation 126, and is configured to perform

[0028] In an exemplary embodiment, the computer system is a stand-alone computer system such as computer system 500 shown in FIG. 5, a network of distributed computers where at least some of the computers are computer systems such as computer system 500 shown in FIG. 5, or a cloud computing node server such as computer system 500 shown in FIG. 5. In one embodiment, the computer system is a computer system 500 as shown in FIG. 5 that controls the purification of a polymer solution via at least a real-time multi-angle light scattering script or computer software application that executes the operations of the method. In one embodiment, the computer system is a computer system / server 512 as shown in FIG. 5 that controls the purification of a polymer solution via at least a real-time multi-angle light scattering script or computer software application that executes the operations of the method. In one embodiment, the computer system is a processing unit 516 as shown in FIG. 5 that controls the purification of a polymer solution via at least a real-time multi-angle light scattering script or computer software application that executes the operations of the method. In one embodiment, the computer system is a machine learning computer software / program / algorithm that performs the control of the purification of a polymer solution via real-time multi-angle light scattering.

[0029] In one embodiment, the computer system is a computer system 500 as shown in FIG. 5 that controls the purification of a polymer solution via a real-time multi-angle light scattering script or computer software application that executes at least operations 110, 112, 114, 116, 118, 120, 122, and 124. In one embodiment, the computer system is a computer system / server 512 as shown in FIG. 5 that controls the purification of a polymer solution via a real-time multi-angle light scattering script or computer software application that executes at least operations 110, 112, 114, 116, 118, 120, 122, and 124. In one embodiment, the computer system is a processing unit 516 as shown in FIG. 5 that controls the purification of a polymer solution via a real-time multi-angle light scattering script or computer software application that executes at least operations 110, 112, 114, 116, 118, 120, 122, and 124.

[0030] Use of MCLUT In a further embodiment, the computer-implemented method, system, and computer program product are as follows: (1) A computer system receives, from a second set of devices, a second baseline MALS signal value and a second baseline UV signal value of a pure buffer flowing from a second chromatographic purification system to the second set of devices, where the second set of devices includes a second MALS device and a second UV detector; (2) In response to receiving the second baseline MALS signal value from the second MALS device, (a) the computer system receives, from the second MALS device, second scattering intensity values of a second sample solution flowing from the second chromatographic purification system to the second MALS device over a second time series, where the second sample solution includes at least one type of polymer, (b) the computer system calculates an average of the second baseline MALS signal values, (c) the computer system subtracts the average of the second baseline MALS signal values from the second scattering intensity values to obtain an excess scattering intensity value Iscatt2 of the second intensity time series; (3) In response to receiving the second baseline UV signal value from the second UV detector, (a) the computer system receives, from the second UV detector, second UV absorption values of the second sample solution flowing from the second chromatographic purification system to the second UV detector over a second time series, (b) the computer system calculates an average of the second baseline UV signal values, (c) the computer system executes a set of logical operations that apply a second UV absorption alignment correction for a second UV absorption alignment parameter and a second UV absorption band broadening correction for a second UV absorption band broadening parameter to the second UV absorption values over the second time series for the second UV detector and the second MALS device to obtain second corrected UV absorption values over the second time series, (d) the computer system subtracts the average of the second baseline UV signal values from the second corrected UV absorption values to obtain an excess UV absorption value of the second UV time series; (4) The computer system determines a second concentration value c2 of the second sample solution and a second absorption coefficient of the second sample solution with respect to the excess UV absorption value of the second UV time series.Calculating according to Beer's law, and (5) receiving, by a computer system, a second pH value of a second sample solution over a time series from a second pH detector and a second conductivity value of the second sample solution over a time series from a second conductivity detector, wherein the second set of devices further comprises a second pH detector and a second conductivity detector; receiving; (6) performing, by the computer system, a set of logical operations to: (a) apply a second pH alignment correction for a second pH alignment parameter to the second pH value over a second time series for the second pH detector and the second MALS device to obtain a second corrected pH value over the second time series; and (b) apply a second conductivity alignment correction for a second conductivity alignment parameter to the second conductivity value over a second time series for the second conductivity detector and the second MALS device to obtain a second corrected conductivity value over the second time series, thereby obtaining a second plurality of signals including an excess scattering intensity value Iscatt2 of a second intensity time series, a second concentration value c2 of the second sample solution, the second corrected pH value, and the second corrected conductivity value; (7) retrieving, by the computer system, from the MCLUT, an aggregate content value %agg and a weight average molar mass M corresponding to the second plurality of signals via at least one of correlation, interpolation, and extrapolation; (8) performing, by the computer system, a set of logical operations to determine whether at least one characteristic of the second sample solution has reached an aggregate content limit with respect to at least one of the retrieved aggregate content value %agg, the retrieved weight average molar mass M, and the second concentration value c2 of the second sample solution; and (9) in response to determining that at least one characteristic of the second sample solution has reached the aggregate content limit, transmitting, by the computer system, a waste diversion command to divert the sample solution to a waste container. In one embodiment, the second absorption coefficient of the sample solution is a characteristic of the sample solution. In one embodiment, the second absorption coefficient is received from user input.,

[0031] In one embodiment, retrieving comprises, by a computer system, retrieving from the MCLUT, via interpolation, an aggregate content value, %agg, and a weight-average molar mass M corresponding to a second plurality of signals when Iscatt2, c2, a second pH value, and a second conductivity value are within the range of values in the MCLUT. In one embodiment, retrieving comprises, by a computer system, retrieving from the MCLUT, via extrapolation, an aggregate content value, %agg, and a weight-average molar mass M corresponding to a second plurality of signals when Iscatt2, c2, a second pH value, and a second conductivity value are outside the range of values in the MCLUT.

[0032] Reaching the aggregate content limit In one embodiment, determining that at least one characteristic of the second sample solution has reached the aggregate content limit comprises, by a computer system, determining that the retrieved aggregate content value %agg exceeds a maximum aggregate content value. In one embodiment, determining that at least one characteristic of the second sample solution has reached the aggregate content limit comprises, by a computer system, determining that the retrieved weight-average molar mass M exceeds a maximum weight-average molar mass value.

[0033] In one embodiment, determining that at least one characteristic of the second sample solution has reached the aggregate content limit comprises (a) a subset %agg of the aggregate content values retrieved from the MCLUT by a computer system k and a subset c2 of the second concentration values k with respect to which, according to the following formula, a cumulative aggregate content value %agg accumulated is calculated, where %agg accumulated =(Σ k %agg k ×c2 k ) / Σ k c2 k wherein, %agg k and c2 kcalculating, determined from the effective start of elution from the second chromatographic purification system, and (b) a cumulative content value %agg by a computer system accumulated executing a set of logical operations for determining that accumulated exceeds the maximum cumulative aggregate content value. In one embodiment, the effective start of elution from the second chromatographic purification system is when the sum of %agg accumulated starts.

[0034] Start of elution In one embodiment, the effective start of elution of the second sample solution from the second chromatographic purification system corresponds to the reception time of a trigger signal from the second chromatographic purification system by the computer system. In one embodiment, the trigger signal is from at least one of user programming and established methods. For example, such a trigger signal can be used in binding and elution gradient ion exchange chromatography, and it is known a priori that a specific time difference between the start of the gradient and the elution of the product contains only the desired protein and has at most a small %agg.

[0035] In one embodiment, the effective start of elution of the second sample solution from the second chromatographic purification system corresponds to the time when the concentration value of the second sample solution reaches the minimum allowable value. In one embodiment, the minimum allowable value is user-defined. For example, the minimum allowable value can be used when the concentration increases in the second chromatographic purification system. Also, for example, the minimum allowable value can be used as a finishing step in flow-through hydrophobic interaction chromatography, where it is known a priori that the pure monomer elutes as the first species at a significant concentration.

[0036] In one embodiment, the effective start of elution of the second sample solution from the second chromatographic purification system corresponds to the time when the weight average molar mass of the second sample solution is within the allowable range of molar mass values. In one embodiment, the allowable range of molar mass values is user-defined. For example, it is empirically known that the species eluting first consists mainly of undesired low molar mass species, and thus it may be advantageous to wait until the weight average molar mass of the solution approaches that of the monomer before diverting the solution to the pool. In ion exchange chromatography, the allowable range of molar mass values can be used. In certain embodiments, the second sample solution contains a protein, and the allowable range of molar mass values corresponds to the molar mass of the monomeric protein in the second sample solution. For example, the molar mass of the monomeric protein in the second sample solution can be determined using size exclusion chromatography, and it is empirically known that the species eluting first consists mainly of undesired aggregates, and thus it may be advantageous to wait until the weight average molar mass of the solution approaches that of the monomer before diverting the solution to the pool.

[0037] In one embodiment, the effective start of elution of the second sample solution from the second chromatographic purification system corresponds to at least two of (a) the time of receipt of a trigger signal from the second chromatographic purification system by a computer system, (b) the time when the concentration value of the second sample solution reaches a minimum allowable value, and (c) the time when the weight average molar mass of the second sample solution is within the allowable range of molar mass values. For example, two correspondences require that multiple conditions be met, and thus can be used in various chromatographic purification processes by reducing the probability of collecting impurities in the pool.

[0038] Monomer MALS signal and MCLUT adjustment In a further embodiment, a computer-implemented method, system, and computer program product include: (1) retrieving, by a computer system, predicted values of Iscatt from MCLUT according to measured values Iscatt_exp1 of c2, conductivity, and pH for a specific portion of a second sample solution flowing from a second chromatographic purification system; (2) calculating, by the computer system, an average value Iscatt_meas1 of Iscatt2 measured for a specific portion of the second sample solution flowing from the second chromatographic purification system; and (3) multiplying, by the computer system, the second time series of excess scattering intensity values Iscatt2 by the ratio of Iscatt_exp1 to Iscatt_meas1 to obtain an adjusted value of Iscatt2. In one embodiment, Iscatt2 is adjusted to account for calibration differences for a first MALS instrument.

[0039] In one embodiment, the second sample solution contains protein, and a specific portion of the second sample solution flowing from the second chromatographic purification system corresponds to monomeric protein. For example, the specific portion corresponding to monomeric protein corresponds when %agg is expected to be equal to zero.

[0040] Computer system In an exemplary embodiment, the computer system is computer system 500 as shown in FIG. 5. Computer system 500 is merely an example of a computer system and does not imply any limitation regarding the use or functionality scope of embodiments of the present invention. In any case, computer system 500 is implemented to execute and / or capable of executing any of the functions / operations of the present invention.

[0041] Computer system 500 includes a computer system / server 512 that can operate with a number of 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 use with computer system / server 512 include, but are not limited to, personal computer systems, server computer systems, thin clients, thick 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 above systems or devices.

[0042] Computer system / server 512 may be described in the general context of computer system-executable instructions, such as program modules, being executed by a computer system. Generally, program modules may include routines, programs, objects, components, logic, and / or data structures that perform particular tasks or implement particular abstract data types. Computer system / server 512 may be implemented in a distributed cloud computing environment where tasks are performed by linked remote processing devices via a communications network. In a distributed cloud computing environment, program modules may be located in both local and remote computer system storage media including memory storage devices.

[0043] As shown in FIG. 5, computer system / server 512 within computer system 500 is shown in the form of a general-purpose computing device. The components of computer system / server 512 may include, but are not limited to, one or more processors or processing units 516, a system memory 528, and a bus 518 that couples various system components including system memory 528 to processor 516.

[0044] Bus 518 represents one or more of several types of bus structures, including a memory bus or memory controller, a peripheral bus, an accelerated graphics port, and a processor or local bus using any of various bus architectures. By way of example and not limitation, such architectures include Industry Standard Architecture (ISA) bus, Micro Channel Architecture (MCA) bus, Enhanced ISA (EISA) bus, Video Electronics Standards Association (VESA) local bus, and Peripheral Component Interconnect (PCI) bus.

[0045] Computer system / server 512 typically includes various computer system readable media. Such media can be any available media that is accessible by computer system / server 512 and includes both volatile and nonvolatile media, removable and non-removable media.

[0046] System memory 528 can 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 further include other removable / non-removable, volatile / non-volatile computer system storage media. By way of example only, storage system 534 can be provided to read from and write to a non-removable non-volatile magnetic medium (not shown, typically called a “hard drive”). Although not shown, a magnetic disk drive for reading from and writing to a removable non-volatile magnetic disk (e.g., a “floppy (registered trademark) disk”), and an optical disk drive for reading from and writing to a removable non-volatile optical disk such as a CD-ROM, DVD-ROM, or other optical media can be provided. In such cases, each can be connected to bus 518 by one or more data media interfaces. As further depicted and described below, memory 528 can include at least one program product having a set (e.g., at least one) of program modules configured to execute the functions / operations of embodiments of the present invention.

[0047] A program / utility 540 having a set (at least one) of program modules 542 can be stored in memory 528, by way of example and not limitation. Exemplary program modules 542 can include an operating system, one or more application programs, other program modules, and program data. Each of the operating system, one or more application programs, other program modules, and program data, or some combination thereof, can include an implementation of a network environment. Program modules 542 generally execute the functions and / or methods of embodiments of the present invention.

[0048] The computer system / server 512 can also communicate with one or more external devices 514, such as a keyboard, a pointing device, a display 524, one or more devices that enable a user to interact with the computer system / server 512, and / or any device that enables the computer system / server 512 to communicate with one or more other computing devices (e.g., a network card, a modem, etc.). Such communication can be carried out via an input / output (I / O) interface 522. Further, 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 depicted, the network adapter 520 communicates with other components of the computer system / server 512 via a bus 518. Although not shown, it should be understood that other hardware and / or software components can be used 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 archival storage systems.

[0049] Computer program product The present invention can be a system, a method, and / or a computer program product. The computer program product can include a computer-readable storage medium (s) having computer-readable program instructions thereon for causing a processor to execute aspects of the present invention.

[0050] A computer-readable storage medium can be a tangible device that can hold and store instructions for use by an instruction execution device. The computer-readable storage medium can be, for example, an electronic storage device, a magnetic storage device, an optical storage device, an electromagnetic storage device, a semiconductor storage device, or any suitable combination of the foregoing, but is not limited thereto. A non-exhaustive list of more specific examples of computer-readable storage media includes portable computer 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 compact disc read-only memory (CD-ROM), digital versatile disk (DVD), memory stick, floppy disk, punch card, or mechanically encoded devices such as raised structures within grooves in which instructions are recorded, and any suitable combination of the foregoing. A computer-readable storage medium, as used herein, should not be construed to be a signal per se, such as radio waves or other freely propagating electromagnetic waves, electromagnetic waves propagating through a waveguide or other transmission medium (e.g., optical pulses passing through an optical fiber cable), or electrical signals transmitted through a wire, which are transient signals themselves.

[0051] The computer-readable program instructions described herein can be downloaded from a computer-readable storage medium to respective computing / processing devices, or to an external computer or external storage device via a network, such as, for example, the Internet, a local area network, a wide area network, and / or a wireless network. The network can include copper transmission cables, optical transmission fibers, 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 transfers the computer-readable program instructions for storage on a computer-readable storage medium within each respective computing / processing device.

[0052] Computer-readable program instructions for carrying out the operations of the present invention may be source code or object code written in any combination of one or more programming languages, including assembly instructions, instruction-set-architecture (ISA) instructions, machine instructions, machine-dependent instructions, microcode, firmware instructions, state-setting data, or object-oriented programming languages such as Smalltalk, C++, and conventional procedural programming languages such as the "C" programming language or similar programming languages. The computer-readable program instructions may be executed entirely on the user's computer, partly on the user's computer, as a stand-alone software package, partly on the user's computer and partly on a remote computer, or entirely on the remote computer or server. In the latter scenario, 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 connections may be made to external computers (e.g., via the Internet using an Internet service provider). In some embodiments, for example, an electronic circuit including a programmable logic circuit, a field-programmable gate array (FPGA), or a programmable logic array (PLA) may execute the computer-readable program instructions by utilizing the state information of the computer-readable program instructions to personalize the electronic circuit for performing aspects of the present invention.

[0053] Aspects of the present invention will be described herein with reference to flowchart illustrations and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of the invention. It will be understood that each block of 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.

[0054] These computer-readable program instructions may be provided to a processor of a general purpose computer, 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 / acts specified in one or more blocks of the flowchart and / or block diagram. These computer-readable program instructions may also be stored in a computer-readable storage medium that can direct a computer, a programmable data processing apparatus, and / or other devices to function in a particular manner, such that the computer-readable storage medium storing the instructions contains an article of manufacture including instructions which implement the function / act specified in one or more blocks of the flowchart and / or block diagram.

[0055] The 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 implement the functions / acts specified in one or more blocks of the flowchart and / or block diagram.

[0056] The flowcharts and block diagrams in the figures 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 block in the flowchart or block diagram may represent a module, segment, or portion of one or more executable instructions for implementing the specified logical function. In some alternative implementations, the functions noted in the blocks may be performed in an order different than that noted in the figures. For example, two blocks shown in succession may, in fact, be executed substantially simultaneously, or the blocks may sometimes be executed in the reverse order depending on the functionality involved. It should also be noted that each block of the block diagrams and / or flowchart illustrations, and combinations of blocks in the block diagrams and / or flowchart illustrations, can be implemented by a dedicated hardware-based system that performs the specified functions or operations, or a combination of dedicated hardware and computer instructions.

[0057] The descriptions of the various embodiments of the present disclosure have been presented for purposes of illustration, but are not intended to be exhaustive or limited to the disclosed embodiments. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The terms used herein were chosen to best explain the principles of the embodiments, the practical application to technologies found in the marketplace, or the technical improvement thereof, or to enable others of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A computer-implemented method, comprising: receiving, by a computer system, a baseline multi-angle light scattering (MALS) signal value and a baseline ultraviolet (UV) signal value of a pure buffer flowing from a chromatography purification system to a set of devices from the set of devices; wherein the set of devices includes a MALS device and a UV detector, and receiving; in response to receiving the baseline MALS signal value from the MALS device, receiving, by the computer system, from the MALS device, scattering intensity values of a sample solution flowing from the chromatography purification system to the MALS device over time; wherein the sample solution includes at least one type of polymer; calculating, by the computer system, an average of the baseline MALS signal values; subtracting, by the computer system, the average of the baseline MALS signal values from the scattering intensity values; The excess scattering intensity value Iscatt of the time series of intensity ｊ is obtained, and in response to receiving the baseline UV signal value from the UV detector, receiving, by the computer system, from the UV detector, UV absorption values of the sample solution flowing from the chromatography purification system to the UV detector over the time series; calculating, by the computer system, an average of the baseline UV signal values; executing, by the computer system, a set of logical operations that apply UV absorption alignment correction for UV absorption alignment parameters and UV absorption band expansion correction for UV absorption band expansion parameters to the UV absorption values over the time series for the UV detector and the MALS device to obtain corrected UV absorption values over the time series; subtracting, by the computer system, the average of the baseline UV signal values from the corrected UV absorption values; obtaining excess UV absorption values of the UV time series; The computer system calculates the concentration value c of the sample solution with respect to the excessive UV absorption value of the UV time series ｊ and the absorption coefficient of the sample solution according to Beer's law, receiving, by the computer system, pH values of the sample solution over the time series from a pH detector, and conductivity values of the sample solution over the time series from a conductivity detector; wherein the set of devices further includes the pH detector and the conductivity detector, and receiving; The computer system applies pH alignment correction for pH alignment parameters to the pH values over the time series for the pH detector and the MALS device to obtain corrected pH values over the time series, and applies conductivity alignment correction for conductivity alignment parameters to the conductivity values over the time series for the conductivity detector and the MALS device to obtain corrected conductivity values over the time series, and executes a set of logical operations, the excess scattering intensity value Iscatt of the intensity time series ｊ the concentration value c of the sample solution ｊ obtaining a plurality of signals including the corrected pH value and the corrected conductivity value The computer system performs a set of logical operations to average each of the plurality of signals over a time range t １ , t ２ ,... t ｎ , which involves executing a set of logical operations to average each of the plurality of signals over the time range The time series includes the time range, The time range corresponds to the fractions of the sample solution collected by a fraction collector connected to the set of devices, Perform the act of executing, Average value, [lscatt, c, pH, cond] １ , [lscatt, c, pH, cond] ２ ,..., [lscatt, c, pH, cond] ｎ to obtain The computer system receives the aggregate content value %agg of the fraction of the sample solution measured by the SEC-MALS device １ , %agg ２ ,... %agg ｎ , and the weight average molar mass value M １ , M ２ ,... M ｎ and The computer system determines the aggregate content value %agg １ , %agg ２ ,... %agg ｎ , and the weight average molar mass value M １ , M ２ ,... M ｎ as the average value [lscatt, c, pH, cond] １ , [lscatt, c, pH, cond] ２ ,..., [lscatt, c, pH, cond] ｎ A computer-implemented method including storing in a polymer characterization look-up table (MCLUT) for ｎ . **Claim 2** The computer system receives, from a second set of devices, a second baseline MALS signal value and a second baseline UV signal value of the pure buffer flowing from a second chromatographic purification system to the second set of devices, The second set of devices includes a second MALS device and a second UV detector, and perform the act of receiving, In response to receiving the second baseline MALS signal value from the second MALS device, The computer system receives, from the second MALS device, second scattering intensity values of a second sample solution flowing from the second chromatographic purification system to the second MALS device over a second time series, The second sample solution contains the at least one type of polymer, The computer system calculates an average of the second baseline MALS signal values, The computer system subtracts the average of the second baseline MALS signal values from the second scattering intensity values to Obtain an excess scattering intensity value Iscatt2 of a second intensity time series, In response to receiving the second baseline UV signal value from the second UV detector, The computer system receives, from the second UV detector, second UV absorption values of the second sample solution flowing from the second chromatographic purification system to the second UV detector over the second time series, The computer system calculates an average of the second baseline UV signal values, The computer system executes a set of logical operations to apply a second UV absorption alignment correction for a second UV absorption alignment parameter and a second UV absorption band broadening correction for a second UV absorption band broadening parameter to the second UV absorption values over the second time series for the second UV detector and the second MALS device, to obtain second corrected UV absorption values over the second time series, The computer system subtracts the average of the second baseline UV signal values from the second corrected UV absorption values, To obtain excess UV absorption values for a second UV time series, The computer system calculates a second concentration value c2 of the second sample solution and a second absorption coefficient of the second sample solution for the excess UV absorption values of the second UV time series according to Beer's law, The computer system receives a second pH value of the second sample solution over the time series from a second pH detector and a second conductivity value of the second sample solution over the time series from a second conductivity detector, The second set of devices further includes the second pH detector and the second conductivity detector, and receives, The computer system applies a second pH alignment correction for a second pH alignment parameter to the second pH values over the second time series for the second pH detector and the second MALS device to obtain second corrected pH values over the second time series, and applies a second conductivity alignment correction for a second conductivity alignment parameter to the second conductivity values over the second time series for the second conductivity detector and the second MALS device to obtain second corrected conductivity values over the second time series, by executing a set of logical operations, To obtain a second plurality of signals including the excess scattering intensity value Iscatt2 of the second intensity time series, the second concentration value c2 of the second sample solution, the second corrected pH value, and the second corrected conductivity value, The computer system retrieves an aggregate content value %agg and a weight average molar mass M corresponding to the second plurality of signals from the MCLUT via at least one of correlation, interpolation, and extrapolation, The computer system executes a set of logical operations to determine whether at least one characteristic of the second sample solution has reached an aggregate content limit with respect to at least one of the taken-out aggregate content value %agg, the taken-out weight average molar mass M, and the second concentration value c2 of the second sample solution. In response to determining that at least one characteristic of the second sample solution has reached the aggregate content limit, the computer system further includes transmitting a waste diversion command to divert the sample solution to a waste container. The method according to claim 1.

3. Said determining that at least one characteristic of the second sample solution has reached the aggregate content limit The method according to claim 2, including the computer system determining that the taken-out aggregate content value %agg exceeds the maximum aggregate content value.

4. Said determining that at least one characteristic of the second sample solution has reached the aggregate content limit The method according to claim 2, including the computer system determining that the taken-out weight average molar mass M exceeds the maximum weight average molar mass value.

5. Said determining that at least one characteristic of the second sample solution has reached the aggregate content limit By the computer system, the aggregate content value %agg extracted from the MCLUT ｋ and a subset of the second concentration value c2 ｋ with respect to a subset of, the cumulative aggregate content value %agg ａｃｃｕｍｕｌａｔｅｄ is to be calculated, %agg ｋ and c2 ｋ is determined from the effective start of elution from the second chromatographic purification system, to calculate, The computer system executes a set of logical operations for determining that the cumulative content value %agg ａｃｃｕｍｕｌａｔｅｄ exceeds the maximum cumulative aggregate content value, the method according to claim 2, comprising.

6. The effective start of the elution of the second sample solution from the second chromatographic purification system corresponds to the reception time of the trigger signal from the second chromatographic purification system by the computer system. The method according to claim 5.

7. The effective start of the elution of the second sample solution from the second chromatographic purification system corresponds to the time when the concentration value of the second sample solution reaches the minimum allowable value. The method according to claim 5.

8. The effective start of the elution of the second sample solution from the second chromatographic purification system corresponds to the time when the weight average molar mass of the second sample solution is within the allowable range of molar mass values. The method according to claim 5.

9. The second sample solution contains protein. The molar mass value within the allowable range corresponds to the molar mass of the monomer protein in the second sample solution. The method according to claim 8.

10. The effective start of the elution of the second sample solution from the second chromatographic purification system corresponds to at least two of (a) the reception time of a trigger signal from the second chromatographic purification system by the computer system, (b) the time when the concentration value of the second sample solution reaches a minimum allowable value, and (c) the time when the weight average molar mass of the second sample solution is within an allowable range of molar mass values, according to the method of claim 5.

11. Retrieving, by the computer system, from the MCLUT, an expected value Iscatt_exp1 of Iscatt according to measured values of c2, conductivity, and pH for a specific portion of the second sample solution flowing from the second chromatographic purification system; Calculating, by the computer system, an average value Iscatt_meas1 of Iscatt2 measured for the specific portion of the second sample solution flowing from the second chromatographic purification system; Multiplying, by the computer system, the second time series of excess scattering intensity values Iscatt2 by the ratio of Iscatt_exp1 to Iscatt_meas1, to obtain an adjusted value of Iscatt2, further comprising the method of claim 2.

12. The second sample solution contains a protein, The specific portion of the second sample solution flowing from the second chromatographic purification system corresponds to a monomeric protein, according to the method of claim 11.

13. A computer-implemented method, receiving, by a computer system, from a set of devices, a baseline MALS signal value and a baseline UV signal value of a pure buffer solution flowing from a chromatographic purification system to the set of devices, the set of devices including a MALS device and a UV detector, receiving; in response to receiving the baseline MALS signal value from the MALS device, receiving, by the computer system, from the MALS device, scattering intensity values of a sample solution flowing from the chromatographic purification system to the MALS device over time, the sample solution containing the at least one type of polymer, calculating, by the computer system, an average of the baseline MALS signal values, The computer system subtracts the mean of the baseline MALS signal values from the scattering intensity values to obtain the excess scattering intensity value Iscatt of the intensity time series, in response to receiving the baseline UV signal value from the UV detector, the computer system receives, from the UV detector, the UV absorption values of the sample solution flowing from the chromatography purification system to the UV detector over the time series, the computer system calculates the mean of the baseline UV signal values, the computer system executes a set of logical operations that apply UV absorption alignment correction for UV absorption alignment parameters and UV absorption band expansion correction for UV absorption band expansion parameters to the UV absorption values over the time series for the UV detector and the MALS device to obtain corrected UV absorption values over the time series, the computer system subtracts the mean of the baseline UV signal values from the corrected UV absorption values to obtain the excess UV absorption value of the UV time series, the computer system calculates the concentration value c of the sample solution and the absorption coefficient of the sample solution for the excess UV absorption value of the UV time series in accordance with Beer's law, the computer system receives the pH value of the sample solution over the time series from a pH detector and the conductivity value of the sample solution over the time series from a conductivity detector, wherein the set of devices further includes the pH detector and the conductivity detector, receiving, the computer system applies pH alignment correction for pH alignment parameters to the pH values over the time series for the pH detector and the MALS device to obtain corrected pH values over the second time series, and applies conductivity alignment correction for conductivity alignment parameters to the conductivity values over the time series for the conductivity detector and the MALS device to obtain corrected conductivity values over the time series, by executing a set of logical operations, obtaining a plurality of signals including the excess scattering intensity value Iscatt of the intensity time series, the concentration value c of the sample solution, the corrected pH value, and the corrected conductivity value The computer system retrieves the aggregate content value %agg and the weight average molar mass M corresponding to the plurality of signals from a polymer characterization lookup table (MCLUT) via at least one of correlation, interpolation, and extrapolation. The computer system executes a set of logical operations to determine whether at least one characteristic of the sample solution has reached an aggregate content limit with respect to at least one of the retrieved aggregate content value %agg of the second sample solution, the retrieved weight average molar mass M, and the concentration value c. In response to determining that the at least one characteristic of the sample solution has reached the aggregate content limit, the computer system transmits a waste diversion command to divert the sample solution to a waste container. A computer-implemented method including.

14. A system comprising a memory and a processor communicating with the memory, the processor receiving a baseline multi-angle light scattering (MALS) signal value and a baseline ultraviolet (UV) signal value of a pure buffer flowing from a chromatographic purification system to the set of devices from the set of devices, the set of devices includes a MALS device and a UV detector, and in response to receiving the baseline MALS signal value from the MALS device, receiving, over time series, the scattering intensity values of the sample solution flowing from the chromatographic purification system to the MALS device from the MALS device, the sample solution includes at least one type of polymer, the computer system calculates an average of the baseline MALS signal values, the computer system subtracts the average of the baseline MALS signal values from the scattering intensity values, The excessive scattering intensity value Iscatt of the intensity time series ｊ is obtained, and in response to receiving the baseline UV signal value from the UV detector, the computer system, from the UV detector, receiving the UV absorption values of the sample solution flowing from the chromatographic purification system to the UV detector over the time series, the computer system calculates an average of the baseline UV signal values, The computer system executes a set of logical operations that apply UV absorption alignment correction for UV absorption alignment parameters and UV absorption band broadening correction for UV absorption band broadening parameters to the UV absorption values over the time series for the UV detector and the MALS device, to obtain corrected UV absorption values over the time series. The computer system subtracts the average of the baseline UV signal values from the corrected UV absorption values. To obtain excess UV absorption values for the UV time series. The computer system calculates the concentration value c of the sample solution regarding the excessive UV absorption value of the UV time series ｊ and the absorption coefficient of the sample solution according to Beer's law, The computer system receives the pH value of the sample solution over the time series from the pH detector and the conductivity value of the sample solution over the time series from the conductivity detector. The set of devices further includes the pH detector and the conductivity detector, and receives. The computer system executes a set of logical operations that apply pH alignment correction for pH alignment parameters to the pH values over the time series for the pH detector and the MALS device to obtain corrected pH values over the time series, and apply conductivity alignment correction for conductivity alignment parameters to the conductivity values over the time series for the conductivity detector and the MALS device to obtain corrected conductivity values over the time series. The excess scattering intensity value Iscatt of the intensity time series ｊ , the concentration value c of the sample solution ｊ , obtaining a plurality of signals including the corrected pH value and the corrected conductivity value The computer system performs a set of logical operations to average each of the plurality of signals over a time range t １ , t ２ ,... t ｎ , wherein the time series includes the time range, and the time range corresponds to a fraction of the sample solution collected by a fraction collector connected to the set of devices, and performs the execution to obtain an average value [lscatt, c, pH, cond] １ , [lscatt, c, pH, cond] ２ ,..., [lscatt, c, pH, cond] ｎ to obtain The computer system determines the aggregate content value %agg of the fraction of the sample solution measured by the SEC-MALS instrument １ , %agg ２ ,... % agg ｎ and the weight-average molar mass value, M １ M ２ M... ｎ to receive The computer system determines the aggregate content value %agg １ , %agg ２ ,... %agg ｎ , and the weight-average molar mass value M １ , M ２ ,... M ｎ as the average value [lscatt, c, pH, cond] １ , [lscatt, c, pH, cond] ２ ,..., [lscatt, c, pH, cond] ｎ A system configured to perform a method that includes storing, in a macromolecule characterization look-up table (MCLUT), for ｎ .

Images