Pharmaceutical formulations in bioproduction

Abstract

A method for mixing solutions includes receiving a flowing first solution containing a biopharmaceutical and directing the flowing first solution along a flow path including a dilution position where a dilution device is in fluid communication with the flow path; introducing a second solution into the flowing first solution at the dilution position to form a flowing third solution; measuring the biopharmaceutical in the flowing first solution at a position upstream from the dilution position; measuring the biopharmaceutical in the flowing third solution at a position downstream from the dilution position; determining a relative relationship between the measurements of the biopharmaceutical or values ​​derived from the measurements of the biopharmaceutical at the upstream and downstream positions; and adjusting the flow rate of at least one of the first and second solutions based on the relative relationship.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 414,452, filed October 7, 2022, the entire contents of which are incorporated herein by reference.

[0002] The present disclosure relates to the production of pharmaceutical formulations in a variety of manufacturing processes, including continuous biomanufacturing. [Background technology]

[0003] Continuous bioproduction processes can be used to produce pharmaceutical products with a wide range of therapeutic effects. Typically, such pharmaceutical products are obtained in solution after production in a bioreactor and, optionally, after one or more purification or other steps have been performed. The final pharmaceutical product is then formulated according to established specifications, which typically includes adjusting the concentrations of the pharmaceutical product and optionally other components in the solution to meet previously established specifications. Excipient solutions can be used to adjust the pharmaceutical solution to meet specifications. Summary of the Invention [Means for solving the problem]

[0004] Certain existing batch pharmaceutical formulation processes use a volumetric flow-based approach to adding excipient solutions. Typically, such processes use a single flow meter or other volumetric measuring device to control the rate of excipient solution addition during the formulation step. In contrast, the present disclosure features methods and systems in which measurements of a biological production process solution are performed multiple times. In particular, measurements are performed both upstream and downstream from locations along the solution flow path, and the upstream and downstream measurements are used to control the addition of excipient solutions to ensure that the final pharmaceutical formulation meets previously established specifications for one or more pharmaceuticals in the process solution. The use of both upstream and downstream measurements can improve pharmaceutical quality, and in particular, the methods and systems described herein are compatible with various pharmaceutical purification steps that can be difficult to implement in traditional pharmaceutical formulation workflows due to the relatively small volumetric flow rates of pharmaceutical-containing solutions and the time-dependent fluctuations in the composition of the pharmaceutical-containing solutions.

[0005] In a first aspect, the disclosure features a method including receiving a flowing first solution containing a biopharmaceutical and directing the flowing first solution along a flow path including a dilution location where a dilution device is in fluid communication with the flow path; introducing a second solution into the flowing first solution at the dilution location to form a flowing third solution; measuring the biopharmaceutical in the flowing first solution at a location upstream from the dilution location; measuring the biopharmaceutical in the flowing third solution at a location downstream from the dilution location; determining a relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream locations; and adjusting a flow rate of at least one of the first and second solutions based on the relative relationship.

[0006] Embodiments of the methods of the present invention may include any one or more of the following features.

[0007] Measuring the biopharmaceutical at the upstream location may include obtaining a measured value of a parameter of the first solution, which may be the refractive index of the first solution, the conductivity of the first solution, the absorbance of the first solution, the transmittance of the first solution, the reflectance of the first solution, and / or the concentration of the biopharmaceutical in the first solution.

[0008] The parameter measurement may be taken at a single wavelength, which may be in the ultraviolet, visible, or infrared region of the spectrum.

[0009] Measuring the biopharmaceutical at the upstream location may include obtaining a plurality of measurements of the first solution. The plurality of measurements may correspond to spectral information of the first solution at a plurality of wavelengths. Each method may include analyzing the spectral information to determine a value derived from the spectral information. Analyzing the spectral information may include using a calibrated chemometric model to determine the value derived from the spectral information.

[0010] The value derived from the spectral information may be the concentration of the biopharmaceutical in the first solution or a quantity related to the concentration of the biopharmaceutical in the first solution. The spectral information may include an infrared spectrum of the first solution and / or an ultraviolet spectrum of the first solution and / or a Raman scattering spectrum of the first solution.

[0011] Each method may include measuring the attenuated total reflectance of incident infrared light to obtain an infrared spectrum. Each method may include measuring the refractive index of the first solution by measuring the attenuated total reflectance of incident infrared light from the first solution.

[0012] Measuring the biopharmaceutical at a downstream location can include obtaining a measurement of a parameter of the third solution, which can include at least one member of the group consisting of a refractive index of the third solution, a conductivity of the third solution, an absorptivity of the third solution, a transmittance of the third solution, a reflectance of the third solution, and a concentration of the biopharmaceutical in the third solution.

[0013] The measurement of the parameter of the third solution may be measured at a single wavelength, which may include a wavelength in the ultraviolet, visible, or infrared region of the spectrum.

[0014] Measuring the biopharmaceutical at a downstream location can include obtaining a plurality of measurements of the third solution, where the plurality of measurements can correspond to spectral information of the third solution at a plurality of wavelengths.

[0015] Each method may include analyzing the spectral information to determine a value derived from the spectral information. Analyzing the spectral information may include using a calibrated chemometric model to determine the value derived from the spectral information.

[0016] The value derived from the spectral information may be the concentration of the biopharmaceutical in the third solution or a quantity related to the concentration of the biopharmaceutical in the third solution. The spectral information may include at least one member of the group consisting of an infrared spectrum of the third solution, an ultraviolet spectrum of the third solution, and a Raman scattering spectrum of the third solution. Each method may include measuring the attenuated total reflectance of incident infrared light to obtain the infrared spectrum. Each method may include measuring the refractive index of the third solution by measuring the attenuated total reflectance of incident infrared light from the third solution.

[0017] Measuring the biopharmaceutical at an upstream location may include obtaining at least one of measured parameters and spectral information for a first solution, and measuring the biopharmaceutical at a downstream location may include obtaining at least one of measured parameters and spectral information for a third solution, where the measured parameters and at least one of the spectral information for the first solution may be measured using a different measurement technique than the measured parameters and at least one of the spectral information for the third solution. The measured parameters and at least one of the spectral information for the first solution may include a different type of information than the measured parameters and at least one of the spectral information for the third solution. The measured parameters and at least one of the spectral information for the first solution and the measured parameters and at least one of the spectral information for the third solution may each be independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, biopharmaceutical concentration, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

[0018] Measuring the biopharmaceutical at an upstream location may include obtaining at least one of a measured value of a parameter of a first solution and spectral information for the first solution, and measuring the biopharmaceutical at a downstream location may include obtaining at least one of a measured value of a parameter of a third solution and spectral information for the third solution, wherein the measured value of the parameter of the first solution and at least one of the spectral information for the first solution and the measured value of the parameter of the third solution and at least one of the spectral information for the third solution may be measured using a common measurement technique. The measured value of the parameter of the first solution and at least one of the spectral information for the first solution and the measured value of the parameter of the third solution and at least one of the spectral information for the third solution may each be independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, concentration of the biopharmaceutical, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

[0019] The second solution may be free of a biopharmaceutical. The flow rate of the first solution along the flow path may be less than 2 mL / min (e.g., less than 1 mL / min).

[0020] Determining the relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream locations may include obtaining a first value correlating with the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in a first solution at the upstream location, obtaining a second value correlating with the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in a third solution at the downstream location, calculating a comparison amount based on the first and second values, and adjusting the flow rate of at least one of the first and second solutions based on the comparison amount. The comparison amount may be a ratio of the first and second values. The comparison amount is a mathematical function of the first and second values.

[0021] Each method may include adjusting a flow rate of at least one of the first solution and the second solution until the value of the relative relationship falls within a range of target values. Each method may include adjusting a flow rate of at least one of the first solution and the second solution until the value of the comparison quantity falls within a range of target values.

[0022] The biopharmaceutical may be a protein. The protein may be an antibody, an antibody fragment, or may comprise a portion of an antibody. The biopharmaceutical in the first solution may be a drug substance, and the third solution may be a pharmaceutical product.

[0023] The biopharmaceutical may be a first biopharmaceutical, the dilution location may be a first dilution location, and the flow path may include a second dilution location downstream from the first dilution location, and each method may include introducing a fourth solution into the flowing third solution at a second dilution location downstream from a location where the first biopharmaceutical is measured in the flowing third solution to form a flowing fifth solution; measuring the second biopharmaceutical in the flowing fifth solution at locations upstream and downstream of the second dilution location; determining a relative relationship between the measurements of the second biopharmaceutical or values ​​derived from the measurements of the second biopharmaceutical at the upstream and downstream locations; and adjusting at least one of the flow rates of the third solution and the fourth solution based on the relative relationship with the second biopharmaceutical.

[0024] Measuring the second biopharmaceutical at a location downstream from the second dilution location can include obtaining a measurement of a parameter of the fifth solution, which can include at least one member of the group consisting of a refractive index of the fifth solution, a conductivity of the fifth solution, an absorptivity of the fifth solution, a transmittance of the fifth solution, a reflectance of the fifth solution, and a concentration of the second biopharmaceutical in the fifth solution.

[0025] The measurement of the fifth solution parameter may be measured at a single wavelength, which may include a wavelength in the ultraviolet, visible, or infrared spectral region.

[0026] Measuring the second biopharmaceutical at a location downstream from the second dilution location can include obtaining a plurality of measurements of the fifth solution, wherein the plurality of measurements can correspond to spectral information of the fifth solution at a plurality of wavelengths.

[0027] Each method may include analyzing the spectral information of the fifth solution to determine a value derived from the spectral information. Analyzing the spectral information of the fifth solution may include using a calibrated chemometric model to determine the value derived from the spectral information. The value derived from the spectral information may be the concentration of the second biopharmaceutical in the fifth solution or a quantity related to the concentration of the second biopharmaceutical in the fifth solution.

[0028] The spectral information of the fifth solution may include at least one member of the group consisting of an infrared spectrum of the fifth solution, an ultraviolet spectrum of the fifth solution, and a Raman scattering spectrum of the fifth solution. Each method may include measuring attenuated total reflectance of incident infrared light to obtain the infrared spectrum of the fifth solution. Each method may include measuring the refractive index of the fifth solution by measuring attenuated total reflectance of incident infrared light from the fifth solution.

[0029] The biopharmaceutical may be measured at locations upstream and downstream from the dilution location by measuring different types of spectral information corresponding to each of the first and third solutions, and the measurements of the second biopharmaceutical or values ​​derived from the measurements of the second biopharmaceutical at the upstream and downstream locations may be of different types.

[0030] Each method may include obtaining measurements of the second biopharmaceutical or values ​​derived from measurements of the second biopharmaceutical at upstream and downstream locations using different measurement techniques.

[0031] The fourth solution may be free of the second biopharmaceutical. The fourth solution may be free of the first biopharmaceutical.

[0032] The flowing first solution can be received from a purification unit of the bioproduction system, which can include a tangential flow filtration unit.

[0033] The measured parameter of the first solution can be the osmolality of the first solution. The measured parameter of the third solution can be the osmolality of the third solution. Measuring the biopharmaceutical at an upstream location can include obtaining an osmolality value of the first solution, and measuring the biopharmaceutical at a downstream location can include obtaining an osmolality value of the third solution.

[0034] Each method can include adjusting the flow rate of the second solution based on the relative relationship, which can be a ratio of the osmolality values ​​of the first and third solutions. Embodiments of each method can also include any other feature described herein, and can include any combination of features, including combinations of features described individually in different embodiments, unless otherwise specified.

[0035] In another aspect, the disclosure provides a system including a flow conduit including an inlet, a reservoir connected to the flow conduit at a dilution position, at least one flow regulator connected either between the reservoir and the dilution position or between the inlet and the dilution position, a first sensor disposed on the flow conduit at an upstream position between the inlet and the dilution position, a second sensor disposed on the flow conduit at a downstream position between the outlet and the dilution position, and a controller connected to the first and second sensors and regulators, wherein the first sensor is configured to measure a biopharmaceutical in a flowing first solution entering the inlet, the reservoir is configured to introduce a second solution into the flow conduit at the dilution position to form a flowing third solution, the second sensor is configured to measure the biopharmaceutical in the flowing third solution, and the controller is configured to determine a relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream positions and adjust the at least one regulator to control a flow rate of at least one of the first and second solutions based on the relative relationship.

[0036] Embodiments of the systems of the present invention may include any one or more of the following features.

[0037] The first sensor may be configured to obtain a measurement of a parameter of the first solution. The first sensor may be a refractive index sensor configured to measure the refractive index of the first solution. The first sensor may be a conductivity sensor configured to measure the conductivity of the first solution. The first sensor may be an absorbance sensor configured to measure the absorbance of the first solution. The first sensor may be a transmittance sensor configured to measure the transmittance of the first solution. The first sensor may be a reflectance sensor configured to measure the reflectance of the first solution. The first sensor may be a concentration sensor configured to measure the concentration of a biopharmaceutical in the first solution.

[0038] The first sensor may be configured to measure the value of the parameter at a single wavelength, which may be in the ultraviolet spectral region, or in the visible spectral region, or in the infrared spectral region.

[0039] The first sensor may be configured to obtain a plurality of measurements of the first solution. The plurality of measurements may correspond to spectral information of the first solution at a plurality of wavelengths. The controller may be configured to analyze the spectral information to determine a value derived from the spectral information. The controller may be configured to analyze the spectral information by using a calibrated chemometric model to determine a value derived from the spectral information. The value derived from the spectral information may be a concentration of the biopharmaceutical in the first solution or a quantity related to the concentration of the biopharmaceutical in the first solution.

[0040] The first sensor may be configured to obtain an infrared spectrum of the first solution and / or an ultraviolet spectrum of the first solution and / or a Raman scattering spectrum of the first solution. The first sensor may be configured to measure attenuated total reflection of incident infrared light to obtain the infrared spectrum. The first sensor may be configured to measure the refractive index of the first solution by measuring attenuated total reflection of incident infrared light from the first solution.

[0041] The second sensor may be configured to obtain a measurement of a parameter of the third solution, and may be configured to measure at least one member of the group consisting of a refractive index of the third solution, a conductivity of the third solution, an absorptivity of the third solution, a transmittance of the third solution, a reflectance of the third solution, and a concentration of the biopharmaceutical in the third solution.

[0042] The second sensor may be configured to measure the value of the parameter of the third solution at a single wavelength, which may include a wavelength in the ultraviolet, visible, or infrared spectral region.

[0043] The second sensor may be configured to obtain a plurality of measurements of the third solution. The plurality of measurements may correspond to spectral information of the third solution at a plurality of wavelengths. The controller may be configured to analyze the spectral information to determine a value derived from the spectral information. The controller may be configured to analyze the spectral information by using a calibrated chemometric model to determine a value derived from the spectral information. The value derived from the spectral information may be a concentration of the biopharmaceutical in the third solution or a quantity related to the concentration of the biopharmaceutical in the third solution.

[0044] The second sensor may be configured to acquire spectral information including at least one member of the group consisting of an infrared spectrum of the third solution, an ultraviolet spectrum of the third solution, and a Raman scattering spectrum of the third solution. The second sensor may be configured to measure attenuated total reflection of incident infrared light to acquire the infrared spectrum. The second sensor may be configured to measure the refractive index of the third solution by measuring attenuated total reflection of incident infrared light from the third solution.

[0045] The first and second sensors may be configured to measure the biopharmaceutical in the first and third solutions using different measurement techniques. At least one of the measured parameter of the first solution and the spectral information of the first solution may include a different type of information than at least one of the measured parameter of the third solution and the spectral information of the third solution. At least one of the measured parameter of the first solution and the spectral information of the first solution and at least one of the measured parameter of the third solution and the spectral information of the third solution may each be independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, biopharmaceutical concentration, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

[0046] The first and second sensors may be configured to measure the biopharmaceutical in the first and third solutions using a common measurement technique, and the measured parameter of the first solution and / or the spectral information of the first solution and the measured parameter of the third solution and / or the spectral information of the third solution may each be independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, biopharmaceutical concentration, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

[0047] The second solution may be free of biopharmaceuticals.

[0048] The first and second sensors may be of different types.

[0049] The controller may be configured to determine the relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream locations by obtaining a first value correlating with the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in a first solution, obtaining a second value correlating with the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in a third solution, calculating a comparative amount based on the first and second values, and adjusting at least one regulator to control the flow rate of at least one of the first and second solutions based on the comparative amount. The controller may be configured to calculate the comparative amount as a ratio of the first and second values. The controller may be configured to calculate the comparative amount as a mathematical function of the first and second values.

[0050] The controller may be configured to adjust the at least one regulator to control the flow rate of at least one of the first solution and the second solution until the value of the relative relationship falls within a range of target values. The controller may be configured to adjust the at least one flow regulator to control the flow rate of at least one of the first solution and the second solution until the value of the comparison quantity falls within a range of target values.

[0051] The biopharmaceutical may be a protein, which may be an antibody, an antibody fragment, or may comprise a portion of an antibody.

[0052] The biopharmaceutical in the first solution may be an active pharmaceutical ingredient, and the third solution may be a pharmaceutical product.

[0053] Each system may include a purification unit for use in a bioproduction system, the purification unit being in fluid communication with the inlet. The purification unit may include a tangential flow filtration unit.

[0054] Embodiments of each system may also include any other feature described herein and, unless otherwise specified, may include any combination of features, including combinations of features described separately in different embodiments.

[0055] In a further aspect, the disclosure features a method including receiving a flowing first solution characterized by a biopharmaceutical and directing the flowing first solution along a flow path including a dilution location where a dilution device is in fluid communication with the flow path; introducing a second solution into the flowing first solution at the dilution location to form a flowing third solution; measuring a value of an attribute of the second solution at a location upstream from the dilution location; measuring a value of an attribute of the third solution at a location downstream from the dilution location; determining a relative relationship between the measured attribute values ​​of the second and third solutions; and adjusting a flow rate of the second solution based on the relative relationship.

[0056] Embodiments of each method may include any one or more of the following features.

[0057] The measure of the attribute of the second solution may be the osmolality of the second solution. The measure of the attribute of the second solution may be the refractive index of the second solution. The measure of the attribute of the second solution may be the conductivity of the second solution. The measure of the attribute of the second solution may be the absorbance of the second solution. The measure of the attribute of the second solution may be the transmittance of the second solution. The measure of the attribute of the second solution may be the reflectance of the second solution.

[0058] The measurement of the attribute of the second solution may be measured at a single wavelength. Measuring the value of the attribute of the second solution may include obtaining multiple measurements of the second solution. The multiple measurements may include spectral information of the second solution at multiple wavelengths.

[0059] Each method may include analyzing the spectral information to determine a value of an attribute of the second solution. Analyzing the spectral information may include using a calibrated chemometric model to determine a value of the attribute of the second solution. The value of the attribute may be the osmolality of the second solution.

[0060] The spectral information may include a Raman scattering spectrum of the second solution. The spectral information may include an infrared spectrum of the second solution. The spectral information may include an ultraviolet spectrum of the second solution.

[0061] The measure of the attribute of the third solution may be the osmolality of the third solution. The measure of the attribute of the second solution may be the osmolality of the second solution, and the measure of the attribute of the third solution may be the osmolality of the third solution. The measure of the attribute of the third solution may be at least one member of the group consisting of the refractive index of the third solution, the conductivity of the third solution, the absorptivity of the third solution, the transmittance of the third solution, and the reflectance of the third solution.

[0062] The measurement of the attribute of the third solution may be measured at a single wavelength. Measuring the value of the attribute of the third solution may include obtaining multiple measurements of the third solution. The multiple measurements may include spectral information of the third solution at multiple wavelengths. Each method may include analyzing the spectral information to determine the value of the attribute of the third solution. Analyzing the spectral information may include using a calibrated chemometric model to determine the value of the attribute of the third solution. The value of the attribute may be the osmolality of the third solution.

[0063] The spectral information may include a Raman scattering spectrum of the third solution. The spectral information may include an infrared spectrum of the third solution. The spectral information may include an ultraviolet spectrum of the third solution.

[0064] The values ​​of the attributes of the second and third solutions may be measured using different measurement techniques. The attributes of the second and third solutions whose values ​​are measured may be different.

[0065] The second solution may be free of a biopharmaceutical. The flow rate of the first solution along the flow path may be less than 2 mL / min.

[0066] Determining the relative relationship between the measured attribute values ​​of the second and third solutions may include calculating a comparative quantity between the measured attribute values, which may be a ratio of the measured attribute values, or which may be a mathematical function of the measured attribute values.

[0067] Each method may include adjusting the flow rate of the second solution until the value of the relative relationship falls within a target range. Each method may include adjusting the flow rate of the second solution until the value of the comparison quantity falls within a target range.

[0068] The biopharmaceutical may be a protein. The protein may be an antibody, an antibody fragment, or a portion of an antibody. The biopharmaceutical in the first solution may be a drug substance, and the third solution may be a pharmaceutical product.

[0069] Embodiments of each method may also include any other feature described herein and, unless otherwise specified, may include any combination of features, including combinations of features described separately in different embodiments.

[0070] In another aspect, the disclosure features a system including a flow conduit featuring an inlet, a reservoir connected to the flow conduit at a dilution position, at least one flow regulator connected between the reservoir and the dilution position, a first sensor disposed between the reservoir and the dilution position, a second sensor disposed downstream between an outlet of the flow conduit and the dilution position, and a controller connected to the first and second sensors and the flow regulators, wherein the flow conduit is configured to receive a flowing first solution including a biopharmaceutical through the inlet, the reservoir is configured to introduce a second solution into the flow conduit at the dilution position to form a flowing third solution, the first sensor configured to measure an attribute value of the second solution and the second sensor configured to measure an attribute value of the third solution, and the controller is configured to determine a relative relationship between the measured attribute values ​​of the second and third solutions and adjust the at least one flow regulator to control a flow rate of the second solution based on the relative relationship.

[0071] Embodiments of each system may include any one or more of the following features.

[0072] The first sensor may be a Raman scattering sensor configured to measure Raman scattered light from the second solution. The first sensor may be a refractive index sensor configured to measure the refractive index of the second solution. The first sensor may be a conductivity sensor configured to measure the conductivity of the second solution. The first sensor may be an absorbance sensor configured to measure the absorbance of the second solution. The first sensor may be a transmittance sensor configured to measure the transmittance of the second solution. The first sensor may be a reflectance sensor configured to measure the reflectance of the second solution.

[0073] The first sensor may be configured to obtain a plurality of measurements of the second solution. The plurality of measurements may include spectral information of the second solution at a plurality of wavelengths. The controller may be configured to analyze the spectral information to determine a measured attribute value of the second solution. The controller may be configured to analyze the spectral information by using a calibrated chemometric model to determine the measured attribute value of the second solution from the spectral information. The first sensor may be configured to obtain a Raman scattering spectrum of the second solution.

[0074] The second sensor may be a Raman scattering sensor configured to measure Raman scattered light from the third solution. The second sensor may include at least one member of the group consisting of a refractive index sensor configured to measure the refractive index of the third solution, a conductivity sensor configured to measure the conductivity of the third solution, an absorbance sensor configured to measure the absorbance of the third solution, a transmittance sensor configured to measure the transmittance of the third solution, and a reflectance sensor configured to measure the reflectance of the third solution.

[0075] The second sensor may be configured to obtain a plurality of measurements of the third solution. The plurality of measurements may include spectral information of the third solution at a plurality of wavelengths. The controller may be configured to analyze the spectral information to determine a measured attribute value of the third solution from the spectral information. The controller may be configured to analyze the spectral information by using a calibrated chemometric model to determine the measured attribute value of the third solution. The second sensor may be configured to obtain a Raman scattering spectrum of the third solution.

[0076] The first and second sensors may be configured to measure attribute values ​​of the second and third solutions using different measurement techniques. The attributes of the second and third solutions whose values ​​are measured may be different. The second solution may not contain a biopharmaceutical. The first and second sensors may be of different types.

[0077] The controller may be configured to calculate a comparison amount based on the measured attribute values ​​of the second and third solutions and adjust at least one regulator to control the flow rate of the second solution based on the comparison amount. The controller may be configured to calculate the comparison amount as a ratio of the measured attribute values. The controller may be configured to calculate the comparison amount as a mathematical function of the measured attribute values.

[0078] The controller may be configured to adjust the at least one regulator to control the flow rate of the second solution until the value of the relative relationship falls within a range of target values. The controller may be configured to adjust the at least one flow regulator to control the flow rate of the second solution until the value of the comparison quantity falls within a range of target values.

[0079] The biopharmaceutical may be a protein. The protein may be an antibody, an antibody fragment, or a portion of an antibody. The biopharmaceutical in the first solution may be a drug substance, and the third solution may be a pharmaceutical product.

[0080] Each system may include a purification unit in fluid communication with the inlet. The purification unit may include a tangential flow filtration unit.

[0081] Embodiments of each system may also include any other feature described herein, and may include any combination of features, including combinations of features described separately in different embodiments, unless otherwise specified. As used herein, the terms "excipient" and "excipient solution" refer interchangeably to a substance, typically (but not always) in a liquid state (i.e., a pure liquid, a solution containing one or more solvents and one or more solutes, a homogeneous or heterogeneous suspension of one or more components in one or more solvents), that is generally added to a solution to form a final pharmaceutical solution. Typically, but not always, the solution to which the excipient is added contains one or more pharmaceuticals from a bioproduction step, and the addition of the excipient does not alter the chemical nature of the one or more pharmaceuticals in the final pharmaceutical solution. An excipient may include, for example, one or more substances that aid in the stabilization, packaging, and / or delivery of one or more pharmaceuticals in the final pharmaceutical solution. Typically, but not always, the substances in an excipient do not chemically react with the one or more pharmaceutical agents in the solution to which the excipient is added. Excipients may be added to a pharmaceutical-containing solution, for example, to adjust the properties of one or more pharmaceutical agents in the final pharmaceutical solution to meet established specifications for the final pharmaceutical solution. Examples of substances that may be present in an excipient include, but are not limited to, buffers, preservatives, bulking agents, chelating agents, colorants, stabilizers / scavengers, and solvents.

[0082] As used herein, the terms "regulator" and "flow regulator" refer to a device or element of a device that can be adjusted to regulate the flow of a fluid through a conduit. Many different types of fluid valves are commercially available and can be used as flow regulators. Additionally, a wide variety of pumps can be electronically controlled and have adjustable pump speeds to regulate the flow of fluid therethrough. Also, many other devices that respond to an active control signal (e.g., an electronic signal) can function as a flow regulator. Typically, a flow regulator controls the rate at which fluid passes through it by adjusting the cross-sectional area of ​​an opening in the flow regulator, or by adjusting the pump speed, or both.

[0083] As used herein, a "mathematical function" corresponding to a comparative quantity is a function of two variables that generates an output value that represents the relative magnitude of the two variables. For example, the mathematical function may be a ratio of the values ​​of the two variables, with the ratio value representing the relative magnitude of the two variables. More generally, the mathematical function corresponding to the comparative quantity may have any functional form that generates an output value that can be used as a feedback indicator to adjust the relative flow rates of the fluids, as described in more detail below. Examples of such functions include, but are not limited to, logarithmic functions, exponential functions, power law functions, polynomial functions, hyperbolic functions, and any combination thereof.

[0084] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the subject matter herein, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. Additionally, the materials, methods, and examples are illustrative only and are not intended to be limiting of the present invention.

[0085] The details of one or more implementations are set forth in the accompanying drawings and the description below. Other features and advantages will be apparent from the description, drawings, and claims. [Brief explanation of the drawings]

[0086] [Figure 1] FIG. 1 is a schematic diagram showing a method for adding excipients to a drug-containing solution in a controlled manner. [Figure 2] FIG. 1 is a schematic diagram illustrating another method for adding excipients to a drug-containing solution in a controlled manner. [Figure 3] FIG. 1 is a schematic diagram of an exemplary system for formulating a pharmaceutical product in a bioproduction operation. [Figure 4] FIG. 1 is a flow diagram illustrating an exemplary series of steps that can be performed to implement controlled pharmaceutical formulation. [Figure 5] FIG. 1 is a schematic diagram of another exemplary system for formulating a pharmaceutical product in a bioproduction operation. [Figure 6] FIG. 1 is a schematic diagram of an example of a multi-stage system for formulating pharmaceutical products in a bioproduction operation. [Figure 7] 1 is a graph showing pharmaceutical formulation behavior over a portion of a 30-day period. [Figure 8] 1 is a graph showing the variation of several measured quantities over time during the execution of a pharmaceutical formulation process. [Figure 9A] 1 is a graph showing measured protein drug concentration in the final formulation as a function of time using two different excipient addition control strategies. [Figure 9B] 1 is a graph showing osmolality measurements as a function of time for final formulations using two different excipient control strategies. [Figure 10] FIG. 2 is a schematic diagram of an example controller of the system described herein. DETAILED DESCRIPTION OF THE INVENTION

[0087] Like reference symbols in the various drawings indicate like elements.

[0088] Preface Batch manufacturing operations have traditionally been used to provide a wide range of biopharmaceuticals, including therapeutic agents and other drugs. More recently, continuous bioproduction processes have replaced traditional batch methods, as continuous operations offer significant improvements over batch processes. For example, continuous bioproduction operations can generally be performed in smaller vessels than batch operations to achieve the same drug yield over time. The smaller processing volumes handled by such vessels allow for shorter mixing times and / or the elimination of mixing hardware, such as impellers, from the vessel. This, in turn, can lead to improved drug product quality due to the reduction in vortices and mechanical shear forces caused by the absence of mechanical disturbances to the solution within the production vessel that would occur if mixing hardware were present.

[0089] Continuous biomanufacturing operations also result in shorter residence times of pharmaceutical products in vessels compared to batch operations. Quality and other attributes of pharmaceutical products and production conditions can be measured in real time or near real time, and this information can be used to dynamically adjust conditions during production operations in ways that may be more difficult or inconsistent in batch operations. As a result, pharmaceutical product yields can be improved, and pharmaceutical lots can be produced in a more predictable and consistent manner.

[0090] Bioproduction operations typically involve a complex series of operations. Pharmaceutical products are produced in bioreactors and then extracted for further processing. Post-extraction processing steps typically include purification in multi-column chromatography systems, buffer and salt adjustment, further polishing via multi-column chromatography, single or multiple stage tangential filtration, and one or more ultrafiltration and diafiltration steps.

[0091] An important aspect of a continuous bioproduction operation generally involves the resulting pharmaceutical formulation after the above-described stages have been performed. Pharmaceutical formulation generally involves adjusting the properties of the pharmaceutical-containing solution to meet established specifications for the pharmaceutical solution to produce a final pharmaceutical product (e.g., a formulated drug substance). For example, the pharmaceutical-containing solution can be adjusted to match the concentration of one or more pharmaceuticals in the solution to the specifications for the solution in the final formulation. Alternatively, or additionally, the pharmaceutical-containing solution can be adjusted to match the concentration of one or more additional substances in the final pharmaceutical solution, such as, but not limited to, bulking agents, buffers, chelating agents, preservatives, colorants, stabilizers / scavengers, and other delivery agents. Further alternatively, or additionally, the pharmaceutical-containing solution can be adjusted to match one or more physical or chemical properties of the solution, such as, but not limited to, pH, viscosity, phase properties, osmolality, and surface tension, to meet specifications.

[0092] Pharmaceutical formulation processes typically include adding one or more excipient solutions ("excipients") to a drug-containing solution to form a final drug solution. Excipients may include substances intended for addition to the final drug solution to adjust the concentration of the substance and / or the properties of the final drug solution. Excipients may include one or more solvents to adjust the concentration of the drug in the final drug solution. For example, by increasing the volume of solvent through the addition of excipients, the concentration of the drug in the final drug solution can be reduced to meet established specifications for the drug formulation.

[0093] In continuous biomanufacturing operations, the pharmaceutical formulation process occurs continuously, i.e., the pharmaceutical-containing solution is adjusted on the fly to produce a final pharmaceutical solution. Various methods can be used to determine the amount of excipient to add to the pharmaceutical-containing solution to produce a final pharmaceutical solution that meets target specifications. For example, some methods use volumetric flow measurements to adjust the addition of excipients to the pharmaceutical-containing solution.

[0094] Figure 1 is a schematic diagram illustrating a method for adding excipients to a drug-containing solution in a controlled manner. In Figure 1, drug-containing solution 112 is obtained from a purification unit 102 (e.g., a single-pass tangential flow filtration unit). Drug-containing solution 112 flows through a flow meter 104, which measures the volumetric flow rate of solution 112. Solution 112 enters a fluid junction 118.

[0095] An excipient 114 is stored in a reservoir 110 and is pumped by a pump 108 through a second flow meter 106. Flow measurements from the flow meters 104, 106 are used to adjust the rate at which the pump 108 pumps the excipient 114. The excipient 114 is transferred to a fluid junction 118 where it combines with the drug-containing solution 112 to form a final drug solution 116.

[0096] 1, the volumetric flow rate of the drug-containing solution 112 is used to adjust the flow rate of the excipients 114 to adjust the properties of the final drug solution 116. For example, if the concentration of the drug in solution 112 is to be reduced in the final drug solution 116, then based on the volumetric flow rate of the solution 112, an appropriate amount of excipients is continuously added to the solution 112 to achieve the desired dilution of the solution 112, thereby producing a final drug solution 116 in which the concentration of the drug in the drug solution 116 meets the target drug concentration specification.

[0097] The method illustrated in FIG. 1 is effective when the flow rate of the solution 112 is sufficiently high so that the flow meter 104 can accurately measure the flow rate. However, in certain bioproduction operations, the flow rate of the solution 112 (and the corresponding flow rate of the excipient 114 measured by the flow meter 106) may be sufficiently low that accurate measurement of the flow rate is difficult. For example, if the purification unit 102 is a single-pass tangential flow filtration unit, the flow rate of the solution 112 exiting the filtration unit may be consistently less than 2 mL / min. Under these conditions, the flow rate of the excipient 114 added to the solution 112 may be less than 0.3 mL / min, for example. Currently available flow meters typically have relatively high measurement errors at flow rates below about 1 mL / min. As a result, the method illustrated in FIG. 1 is prone to errors resulting from both the measurement of the flow rate of the solution 112 and the measurement of the flow rate of the excipient 114. These errors make it difficult to accurately add the excipient 114 to the solution 112 to achieve target specifications, potentially resulting in unacceptable variability in some situations. Due to these limitations, unacceptable variations in osmolality and drug concentration in the final drug solution 116 have been observed experimentally.

[0098] An alternative method for regulating the addition of excipients to a drug-containing solution is shown in Figure 2. Certain elements shown in Figure 2 are similar to elements in Figure 1 and are labeled with the same reference numbers. In Figure 2, drug-containing solution 112 flows out of purification unit 102, and the flow rate of solution 112 is measured by flow meter 104. Solution 112 flows through flow meter 104 into fluid junction 118.

[0099] Reservoir 110 contains excipients 114 that are pumped from reservoir 110 by pump 108 and transferred to fluid junction 118. Excipients 114 mix with solution 112 in fluid junction 118 to produce a final drug solution 116 that contains the same drug as solution 112. Based on the flow rate of solution 112 measured by flow meter 104 and the expected concentration of drug in solution 112, an appropriate flow rate for excipients 114 is determined such that the concentration of drug in solution 116 following addition of excipients to solution 112 in junction 118 meets the target specification for solution 116.

[0100] A sensor 202 is positioned to measure the concentration of the drug in the solution 116 following the addition of the excipient 114. The measured concentration of the drug in the solution 116 is then used to adjust the flow rate of the excipient 114 via adjustment of the pump 108. In this manner, controlled adjustments of the excipient flow rate can be made to more precisely achieve the target concentration of the drug in the solution 116.

[0101] 2 is based on the assumption that the concentration of the pharmaceutical agent in solution 112 remains relatively constant over time as the solution 112 flows out of filtration unit 102. In reality, this assumption may not hold in many situations. In particular, the concentration of the pharmaceutical agent in solution 112 may change significantly over time due to upstream changes in other processing and / or purification steps. It has been observed experimentally that in some situations, a single sensor 202 may be insufficient to reliably achieve the target specification for the pharmaceutical agent concentration in solution 116, even when the pharmaceutical agent concentration in solution 112 fluctuates significantly.

[0102] This disclosure features several methods and systems that can be used to improve control of pharmaceutical formulation steps in continuous biomanufacturing operations. Specifically, each method and system uses a flow-based control strategy and multiple sensor measurements to mitigate the impact of flow rate and drug concentration fluctuations on excipient addition control. By measuring the drug-containing solution both before and after excipient addition, the resulting control scheme can dynamically adapt to a wide range of process variations. Furthermore, by using many different types of sensors, a robust control strategy can be implemented to reliably achieve many different target specifications for the pharmaceutical formulation.

[0103] Pharmaceutical preparation control method and system 3 is a schematic diagram illustrating an example of a system 300 configured to formulate a drug-containing solution into a final drug solution that achieves target specifications (i.e., one or more target properties) for the final drug solution. The system includes a flow meter 302, a first sensor 304, a fluid junction 306, a second sensor 308, a pump 310, a reservoir 312, and a controller 326. The controller 326 is connected to and in communication with the flow meter 302, the sensors 304, 308, and the pump 310. Conduits 314, 316, and 318 establish a flow path for the solution within the system 300.

[0104] As shown in Figure 3, system 300 is configured to add excipients to a drug-containing solution as described above. The excipients (represented by arrow 322) are contained in reservoir 312 and are transported by pump 310 to fluid junction 306. The drug-containing solution (represented by arrow 320) enters system 300 from an upstream source (e.g., a filtration unit, a conduit, or another element or stage of a continuous biological production system) and is mixed with excipients 322 at fluid junction 306. The resulting final drug solution (represented by arrow 324) flows from fluid junction 306 into a conduit from which it can be further transported to another portion of the continuous biological production system, undergo quality control validation, or be packaged.

[0105] The system 300 includes two sensors 304, 308 located upstream and downstream, respectively, of the location where the solution 320 is mixed with the excipients 322. During operation of the system 300, a controller 326 receives measurements from the flow meter 302 and the sensors 304, 306 and determines the appropriate flow rate of the excipients 322 to ensure that the attributes of the final pharmaceutical solution 324 meet the target specifications for the final pharmaceutical solution. The controller 326 sends control commands to adjust the pump 310, thereby controlling the rate at which the excipients 322 are transported to the fluid junction 306 to be mixed with the solution 320.

[0106] As explained above, the addition of excipients in continuous bioproduction processes generally occurs after the drug product has been produced and purified as part of the drug product formulation process. The specific properties of the excipients depend on the specifications of the final drug product. In a typical drug product formulation process, for example, excipients include concentrated buffer solutions added in amounts sufficient to meet the target specifications of the final drug product. Because continuous bioproduction processes typically produce drug-containing solutions at relatively low flow rates, excipient addition generally occurs at low flow rates (e.g., 10–20% of the flow rate of the drug-containing solution 320). As discussed above, flow meter errors are likely to occur at typical flow rates in such processes, and volumetric flow-based control methods can be inaccurate. Furthermore, the components and concentrations of the drug-containing solution may fluctuate over time, and single-measurement feedback control strategies for excipient addition often fail to adequately respond to such fluctuations, potentially resulting in drug product formulations that are out of specification.

[0107] In contrast, the control scheme implemented by controller 326 uses both pre-formulation and post-excipient addition measurements of the solution (i.e., upstream and downstream from fluid junction 306) to dynamically control the flow rate of excipients 322, ensuring that the final drug solution 324 meets target specifications even if the properties of drug-containing solution 320, such as the concentration of drug in the solution, vary over time, for example, as upstream process conditions change. The drug formulation process implemented by system 300 responds automatically to such changes.

[0108] Notably, because sensors 304, 308 are not flow meters, they provide accurate solution measurements even when the solution flow rate is very low (or when the solution is not flowing at all). Thus, the control scheme described herein is less susceptible to errors that typically result from using flow meters at low solution flow rates. Furthermore, because the control scheme relies on a ratio of measurements, changes in the composition of the pharmaceutical-containing solution are reflected in both measurements, and therefore the ratio of the measurements is not affected by such changes.

[0109] 4 is a flow diagram illustrating an example of steps that can be used to implement the control scheme described herein. In step 402, the flow rate of the drug-containing solution 320 (F) is measured by the flow meter 302. Then, in step 404, the controller 326 receives the measurement information and calculates the flow rate of the excipient (I SP ) is determined according to the following formula:

number

[0110] In step 406, a drug-containing solution 320 (M i ) and final drug solution 324 (M f ) are obtained by sensors 304, 308, respectively. In general, the measurements may correspond to a wide variety of different types of measurements. In some embodiments, for example, the measurements correspond to measurements of concentrations of components in drug-containing solution 320 and final drug solution 324, such as the concentration of a drug in those solutions.

[0111] Controller 326 receives these measurements and determines the ratio of the measurements R in step 408 according to the following equation:

number

[0112] Next, in step 410, the controller 326 determines a value for the adjustment factor α for the flow rate of the excipient 322 based on the ratio of the measured value R to a set point S that represents a desired ratio between the measured values ​​of the drug-containing solution 320 and the final drug solution 324. As an example, if the measured values ​​of the two solutions represent concentrations of components of a solution such as a drug, the set point S represents a desired ratio between the concentrations of the components in the drug-containing solution 320 and the final drug solution 324. The adjustment factor α is therefore calculated according to the following formula:

Number

[0113] Next, in step 412, the controller 426 adjusts the flow rate of the excipient 322 to the fluid junction 306 based on the adjustment coefficient α. To perform this adjustment, the controller 426 calculates the pharmaceutical αI SP and scales the initial set point I SP of the flow rate of the excipient 322 according to the adjustment coefficient. The controller 426 then sends appropriate control commands to the pump 310 to adjust the flow rate of the excipient 322 to this scaled value.

[0114] In the decision step 414, if the pharmaceutical formulation is complete, the control proceeds to step 416 and the procedure ends. However, if the pharmaceutical formulation is not complete, the control returns to step 406, and further measured values of the pharmaceutical-containing solution 320 and the final pharmaceutical solution 324 are obtained by the sensors 304, 308, respectively. A new value of the adjustment coefficient α is obtained, and the flow rate of the excipient 322 is further adjusted based on the new value of the adjustment coefficient. Thus, the control system dynamically responds to changes in the composition and other properties of the pharmaceutical-containing solution 320 as it flows into the system 300.

[0115] In system 300, pump 310 is used as a flow regulator to adjust the flow rate of excipient 322 to junction 306. Pump 310 receives a control signal from controller 326 to adjust the throughput of pump 310. In general, pump 310 may be implemented in the form of any of a variety of adjustable pumps responsive to external control signals. Alternatively, or additionally, the flow regulator may be implemented in system 300 as a controllable valve that receives and responds to a control signal from controller 326. For example, a fixed or variable flow pump may be used as a flow regulator in system 300 in combination with a valve to adjust the rate at which excipient 322 is introduced to junction 306. More generally, any combination of pumps, valves, and other flow restriction and / or flow adjustment elements may be used to adjust the flow rate of excipient 322, provided that the combination provides an adjustable flow rate. In the following discussion, the term "flow regulator" is used to refer to all such combinations of elements that allow for controllable fluid flow.

[0116] In the above description of system 300, controller 326 uses a flow regulator to regulate the flow rate of excipient 322. However, in some embodiments, controller 326 can regulate the flow rate of solution 320 to junction 306 as an alternative to regulating the flow rate of excipient 322. In the control schemes described above, it is the relative flow rates of the drug-containing solution and excipient that are regulated. Such regulation can be accomplished by maintaining the flow rate of one solution constant and adjusting the other.

[0117] A flow regulator (e.g., corresponding to any combination of the elements described above) can be positioned between the system inlet and junction 306 to regulate the flow of solution 320 to junction 306. Figure 5 is a schematic diagram illustrating an example system 500 including a flow regulator 502 positioned to regulate the flow of solution 320 to junction 306. The flow regulator 502 is connected to a controller 326 and adjusts the flow rate of solution 320 in response to control signals from the controller 326. The flow regulator 502 may be implemented as a valve alone (e.g., through which solution 320 is already flowing), as a pump, as a combination of one or more pumps and one or more valves, or as any other combination of components that controllably regulate the flow of solution 320.

[0118] Additionally, in certain embodiments, the systems described herein are capable of actively adjusting the flow rates of both solution 320 and excipient. As discussed above, it is the relative flow rates of these solutions that underpin the control scheme. As such, the methods described herein may be performed by a system that includes a flow regulator (e.g., flow regulator 502 in FIG. 5 ) positioned to regulate the flow of solution 320 to junction 306 and a flow regulator (e.g., where pump 310 is located in FIG. 3 ) positioned to regulate the flow of excipient 322 to junction 306.

[0119] As described above, a variety of different types of measurements can be measured by sensors 304, 308. In some embodiments, the measurements correspond to concentrations or amounts of components of solutions 320, 324 that are not present in excipients. For example, the components may be pharmaceuticals produced during a continuous bioproduction process. Such pharmaceuticals may include, but are not limited to, proteins and fragments thereof, antibodies and fragments thereof, enzymes (e.g., therapeutic enzymes), blood factors, multispecific antibodies, nanobodies, and viral vectors.

[0120] In certain embodiments, the measurements correspond to measurements related to determining the concentration or amount of a component of the solution 320, 324. For example, the measurements may correspond to physical or chemical properties of the solution such as, but not limited to, absorbance, transmittance, reflectance, refractive index, light scattering intensity, electrical conductivity, fluorescence intensity, and Raman scattering intensity related to the presence of the component in the solution 320, 324.

[0121] More generally, in certain embodiments, the measurements correspond to measurements of properties of the solutions 320, 324 for which target specifications have been established in the target formulation. Such properties include, but are not limited to, pH, viscosity, conductivity, and osmolality. Such properties may also include, but are not limited to, pharmaceutical quality attributes, including any of the pharmaceutical quality attributes described in U.S. Patent Application Publication No. 2019 / 0272894, the entire contents of which are incorporated herein by reference. Chemical measurement-based methods, such as those described in U.S. Patent Application Publication No. 2019 / 0272894, can be used to determine the value of the pharmaceutical quality attribute from the corresponding spectral information.

[0122] The sensors 304, 308 may generally be implemented in a variety of different configurations. In certain embodiments, the sensors 304, 308 may be sensors that measure optical and / or non-optical parameters (e.g., absorbance, transmittance, reflectance, fluorescence, refractive index, Raman scattering intensity, conductivity) of the solutions 320, 324. Particular sensors may be configured to measure values ​​of these measurands at a single wavelength (e.g., a single-point measurement) or multiple wavelengths (e.g., a spectral or multi-point measurement). Sensors that perform optical measurements at multiple wavelengths are generally referred to herein as "spectral sensors."

[0123] Sensors configured to measure optical parameters can obtain measurements within a variety of wavelength ranges, including but not limited to the ultraviolet range (e.g., 150 nm to 400 nm), the visible range (e.g., 400 nm to 780 nm), and the infrared range (e.g., 780 nm to 3 μm).

[0124] In some embodiments, sensors 304, 308 are implemented as the same type of sensor. However, more generally, sensors 304, 308 need not be the same type of sensor, but may be implemented as different types of sensors measuring different quantities to obtain information about solutions 320, 324. For example, sensor 304 can measure UV absorbance of solution 320, while sensor 308 can measure fluorescence of solution 324. As another example, sensor 304 can measure infrared absorbance or transmittance at multiple wavelengths, while sensor 308 can measure Raman scattering intensity at multiple wavelengths. As yet another example, sensor 304 can measure refractive index, while sensor 308 can measure UV transmittance at a single wavelength. It will be understood that the above are merely examples, and sensors 304, 308 may be selected from any combination measuring any of the above quantities at one or more wavelengths, whether the same or different, in certain embodiments of the control schemes described herein.

[0125] Suitable sensors for measuring each of the different parameters mentioned above are commercially available. To measure refractive index, sensor 304 and / or sensor 308 may be implemented, for example, as a Pall Ampath reflectometer (available from Pall Corporation, New York, NY). To measure absorbance or transmittance in the ultraviolet region of the spectrum, sensor 304 and / or sensor 308 may be implemented, for example, as an Optek AF46 sensor (available from Optek International, Largo, FL) or as a PendoTECH UV photometer (available from Pendotech, Princeton, NJ). Alternatively, to measure absorbance or transmittance, sensor 304 and / or sensor 308 may be implemented as a variable pathlength spectrometer, such as a FlowVPE system (available from Repligen Corporation, Waltham, MA). To measure Raman scattering intensity, sensor 304 and / or sensor 308 may be implemented as a MarqMetrix system (available from MarqMetrix, Seattle, WA).

[0126] The above examples of sensors are not exhaustive, and in general, the control strategies described herein may be implemented with a wide variety of different sensors that measure many different properties of the solutions 320, 324. Preferably, the sensors used to implement the control strategies take measurements in real time or near real time (e.g., individual measurements are taken within a time window of 30 seconds or less).

[0127] 3 and 4, the flow rate of excipient 322 is adjusted according to measurements obtained from sensors 304 and 308. However, more generally, the flow rate of solution 320 can be adjusted in addition to, or instead of, adjusting the flow rate of excipient 322. A flow regulator, such as an adjustable valve, can be positioned upstream of fluid junction 306 to adjust the flow rate of solution 320, and controller 326 can send appropriate control commands to the flow regulator to adjust the flow rate of solution 320 entering fluid junction 306. By controlling the flow rate of excipient 322, controlling the flow rate of solution 320, or controlling the flow rates of both excipient 322 and solution 320, controller 326 can adjust the properties of final pharmaceutical solution 324. As such, any of these different flow rate adjustments can be used in the control schemes described herein.

[0128] In some embodiments, sensors 304 and / or 308 obtain single-point measurements of solutions 320 and / or 324. Such measurements may correspond directly to physical or chemical properties of the solutions (e.g., such measurements may correspond directly to concentrations of components in the solutions and may be transmitted to controller 326 as concentration values). Alternatively, such measurements may be converted by controller 326 into measurements of physical or chemical properties of the solutions. For example, the physical or chemical properties of the solutions may be calculated by controller 326 as mathematical functions of the measurements of the solutions. The mathematical functions may generally take a wide variety of forms. For example, the value of the physical or chemical property of the solutions may be determined as a linear or nonlinear mathematical function of the measurements obtained for the solutions. The nonlinear functions may be any of a wide variety of different types of functions, including, but not limited to, exponential functions, logarithmic functions, polynomial functions, hyperbolic functions, and combinations of any two or more types of functions.

[0129] In certain embodiments, sensors 304 and / or 308 take multi-point measurements of solutions 320 and / or 324. Such measurements may include, for example, measurements at different times that can be averaged, integrated, or otherwise combined to generate an output measurement that is sent to controller 326. Alternatively, or additionally, such measurements may include measurements at multiple different wavelengths, i.e., spectral measurements.

[0130] When spectral measurements are obtained, a physical or chemical property of the solution can be calculated as a mathematical function of the measurements at different individual wavelengths. Such a mathematical function may include multiple dependent variables corresponding to two or more spectral measurements, and may be a linear or nonlinear function of each of the multiple dependent variables, as described above.

[0131] In some embodiments, when spectral measurements are taken by sensors 304 and / or 308, a chemical measurement-based method can be used to obtain a property value of the solution from which the measurements were taken. Chemical measurement-based methods are generally applicable to a wide variety of different types of measurements and can be used to determine the value of many different types of solution properties. Chemical measurement-based methods are described, for example, in U.S. Patent Application Publication No. 2022 / 0101953, the entire contents of which are incorporated herein by reference.

[0132] 3 and 4 illustrate a control scheme that can be implemented to ensure that the final drug solution 324 meets a target specification for a particular parameter (e.g., drug concentration or amount in solution 324, or another physical or chemical property of solution 324), and it should also be noted that multiple stages of the control scheme can be implemented sequentially to ensure that the output solution meets target specifications for multiple parameters. For example, solution 324 can be directed into another system similar to system 300, allowing solution 324 to function as a drug-containing solution.

[0133] 6 is a schematic diagram of an example system 600 including two stages, each implementing the control scheme described herein. The first stage 650 includes elements similar to those of system 300. In general, the first stage 650 may include any different combination of the features, implementations, and embodiments described above.

[0134] Solution 324 exits first stage 650 and enters second stage 660, which includes fluid junction 606, third sensor 608, pump 610, and reservoir 612 containing second excipient 622. Pump 610 and reservoir 612 are connected to the fluid junction by conduit 618.

[0135] During operation of system 600, first stage 650 functions as described above to produce solution 324 that meets target specifications for a particular property or component (e.g., a first pharmaceutical agent present in solution 324). Solution 324 flows into second stage 660, which also functions as described above but introduces second excipient 622 into solution 324. Sensor 608 measures the solution after addition of excipient 622, and the measurement is transmitted to controller 326. Controller 326 adjusts the relative rates at which excipient 622 and solution 324 flow into fluid junction 606 to ensure solution 624 meets another target specification, i.e., a different target specification than that of first stage 650. As an example, first stage 650 and second stage 660 can operate to ensure that the concentrations of two different pharmaceutical agents initially present in solution 320 meet the target specifications in solution 624. As another example, first stage 650 and second stage 660 may be operable to ensure that the concentration of pharmaceutical agent and other chemical or physical properties of solution 624 meet target specifications.

[0136] In general, both the first stage 650 and the second stage 660 can include any of the elements, implementations, and embodiments described herein, including any of the flow regulators and sensors described (and different combinations thereof). The two stages can regulate any different components and combinations of properties of the incoming solution 320. Furthermore, in certain embodiments, some elements described herein can be omitted. For example, in system 600, sensor 608 functions as a downstream sensor in the second stage 660, while sensor 308 functions as a downstream sensor in the first stage 650 and an upstream sensor in the second stage 660. Alternatively, in some embodiments, the second stage 660 can include another sensor disposed between sensor 308 and fluid junction 606, connected to controller 326 and functioning as an upstream sensor for the second stage 660. This additional sensor can be any of the different types of sensors described herein.

[0137] Each system described herein is not limited to only two stages as shown in Figure 6. In general, each system may include any number of stages (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 8 or more, 10 or more, or more) arranged in series. Multiple target specifications can be met by implementing series of stages similar to system 600.

[0138] Hardware and Software Implementation 10 illustrates an example of a controller 326 that may be used in the systems and methods disclosed herein. The controller 326 may include one or more processors 1002, memory 1004, storage devices 1006, and an interconnecting interface 1008. The processor(s) 1002 may process instructions executed within the controller, including instructions stored in the memory 1004 or storage devices 1006. For example, these instructions may instruct the processor 1002 to perform any of the analysis and control steps disclosed herein.

[0139] The memory 1004 can store executable instructions for the processor 1002, information about system parameters such as excitation and detection wavelengths, and measured image information. The storage device 1006 can be a computer-readable medium such as a floppy disk drive, a hard disk drive, an optical disk drive, or a tape drive, a flash memory or other similar solid-state memory device, or an array of devices including devices in a storage area network or other configuration. The storage device 1006 can store instructions executable by the processor 1002, as described above, and any other information that can be stored in the memory 1004.

[0140] In some embodiments, controller 326 may include a graphics processing unit that displays graphical information (e.g., using a GUI or text interface) on an external input / output device such as display 1016. The graphical information may be displayed by a display device (e.g., a cathode ray tube (CRT) or liquid crystal display (LCD) monitor) that displays any information, such as the measured and calculated spectra and images disclosed herein. A user may provide input to controller 326 using an input device (e.g., a keyboard, pointing device, touch screen, voice recognition device). In some embodiments, one or more of these devices may be part of controller 326.

[0141] A user of any of the systems described herein can provide a variety of different types of commands and information to controller 326 via an input device. The commands and information can include, for example, target specifications for any drug substance produced via the bioproduction operations and systems, information about excipients, measurement information and selection, and calibration information for any elements of the systems and method steps described herein. Controller 326 can use any of these different types of information to perform each of the methods and functions described herein. It should also be noted that any of these types of information can be stored (e.g., in storage device 1006) and recalled as needed by controller 326.

[0142] The methods and individual steps disclosed herein can be performed by the controller 326 by executing instructions in one or more computer programs executable and / or interpretable by the controller 326. These computer programs (also known as programs, software, software applications, or code) include machine instructions for a programmable processor and may be implemented in a high-level procedural and / or object-oriented programming language, and / or an assembly / machine language. For example, a computer program may include instructions stored in the memory 1004, the storage device 1006, and / or a tangible computer-readable medium, as described above, and executed by the processor 1002. As used herein, the term "computer-readable medium" refers to any computer program product, apparatus, and / or device (e.g., magnetic disk, optical disk, memory, programmable logic device (PLD), ASIC, etc.) that includes a machine-readable medium for receiving machine instructions, which is used to provide machine instructions and / or data to a programmable processor.

[0143] By executing the above-described instructions (which may optionally be part of controller 326), the controller may be configured to perform any one or more of the various steps described in connection with any of the control strategies described herein. For example, controller 326 may receive measurements from sensors and flow meters, calculate any of the quantities described herein (and other quantities as well), determine actions based on the quantities and other decision criteria, and send control commands to any of the system elements described herein.

[0144] application The control strategies described herein are particularly applicable to formulation processes in continuous biological production operations, but can also be applied to pharmaceutical formulation processes in other biological production operations, such as batch operations. Furthermore, while the control strategies are described herein in the context of pharmaceutical formulation processes, the strategies can also be applied to additional upstream steps in biological production operations (e.g., prior to pharmaceutical formulation). For example, the strategies can be applied to the addition of salts, solvents, detergents, and other agents, as well as dilution of process fluids, and more generally to any operation or step where target specifications for an output solution or fluid stream are established.

[0145] For example, in some embodiments, the control schemes described herein can be applied to intermediate processes in which a process fluid is diluted to reduce the concentration of a component in the fluid (e.g., a process performed after extraction of a pharmaceutical agent from a bioreactor but prior to pharmaceutical formulation). Any of the various types of measurements described above can be used to adjust the relative flow rates of the fluids that perform the dilution. The measurements can correlate to the pharmaceutical agent in the fluid, or to another non-pharmaceutical species in the fluid, since all components of the fluid are similarly diluted by the addition of another fluid. The fluid added for dilution purposes can include any of the components described herein in connection with excipients, and additional components can be added to the process fluid in addition to diluting the process fluid. For example, the added solution can contain one or more salts such that the process fluid is simultaneously diluted using the methods and systems described herein to increase the salt content of the fluid. As another example, the fluid added to the process fluid can contain a detergent that is added to the process fluid simultaneously with dilution.

[0146] The dilution operation described above can be performed at several different stages of a bioproduction operation. For example, dilution can be used as a load adjustment step during a purification operation to ensure that the chromatography system is sufficiently loaded but not overloaded. Dilution operations can also be implemented as precursors to other unit operations, such as polishing and filtration steps. [Example]

[0147] Example 1: Continuous bioproduction The above control strategy was implemented over a 30-day period in a continuous bioproduction operation carried out in a 500 L bioreactor, where a process stream containing a protein pharmaceutical was produced and the protein concentration in the final pharmaceutical solution was adjusted to meet target specifications.

[0148] Figure 7 is a graph illustrating the performance of the control scheme over a portion of a 30-day period. In Figure 7, trace 702 corresponds to the target concentration of protein in the final pharmaceutical solution, and trace 704 corresponds to the measured protein concentration in the solution. As shown in Figure 7, the measured protein concentration generally meets the target protein concentration until the time corresponding to the vertical line where the measured concentration of protein falls below the target concentration. If the flow rate of the excipient (trace 708) were maintained at a flow rate determined from the volumetric flow measurement (e.g., determined solely based on the flow rate measured by flow meter 302), the excipient flow rate, represented by trace 706, would consistently be too high and the protein concentration in the final pharmaceutical solution would consistently be too low.

[0149] However, by implementing the control scheme described herein, the measured protein concentration was successfully adjusted. At the time indicated by the vertical line in Figure 7, the adjustment coefficient α (trace 710) calculated by controller 326 decreased, causing controller 326 to reduce the flow rate of excipients added to the drug-containing solution. As a result, the measured concentration 704 of protein in the final drug solution increased to meet the target protein concentration 702 within 30 minutes of the initial deviation of the protein concentration from the target concentration.

[0150] FIG. 8 is a graph showing the variation of several measured quantities over time during a pharmaceutical formulation process performed with the control scheme described herein. In FIG. 8, trace 802 shows the measured protein concentration in the solution (e.g., as measured by sensor 304) before the addition of excipients, trace 804 shows the measured protein concentration (e.g., as measured by sensor 308) after the addition of excipients, and trace 806 shows the measured protein concentration in the pharmaceutical formulation. Trace 808 shows the measured flow rate of the drug-containing solution (e.g., as measured by flow meter 302), and trace 810 shows the flow rate of the excipients added to the drug-containing solution. As can be seen from the figure, each of these quantities exhibited measurable fluctuations over time. Nevertheless, as shown by trace 806, which corresponds to the concentration of protein in the pharmaceutical formulation, the control scheme was able to compensate for these fluctuations over time, resulting in a final formulation in which the protein concentration remained substantially constant over the time window shown in the figure.

[0151] Figure 9A is a graph showing measured values ​​of protein drug concentration in the final drug solution as a function of time using a control strategy based solely on the measured volumetric flow rate of the drug-containing solution (solid circles), as described above in connection with Figure 1, and using the control scheme described in connection with Figures 3 and 4 (hatched circles). Figure 9B is a graph showing measured values ​​of osmolality of the final drug solution as a function of time using a control strategy based solely on the measured volumetric flow rate of the drug-containing solution (solid circles), as described above in connection with Figure 1, and using the control scheme described in connection with Figures 3 and 4 (hatched circles). It is clear from the data shown in these figures that, by using the two-sensor control scheme described herein, fluctuations in both protein concentration and osmolality of the final drug solution were significantly reduced, with less than 5% variation over the time window shown, compared to the more than 20% fluctuations in measured protein concentration and osmolality observed when using a control strategy based solely on the measured volumetric flow rate of the drug-containing solution.

[0152] Example 2: Osmotic Pressure-Based Control In this example, osmolality measurements were used to control excipient addition during the bioproduction process. Osmolality is a typical essential quality attribute in a drug product because it can confirm solute content and can be used to detect deviations in the formulation. As used herein, osmolality is defined as the total concentration of all osmotically active dissolved solutes present in a given volume of solution. Osmolality can be measured using an osmometer, which determines the freezing point of an aqueous solution containing one or more dissolved solutes. Osmolality can be determined from measuring the freezing point of a solution because dissolved solutes depress the freezing point of the solution relative to the corresponding pure solvent.

[0153] Conventional osmotic pressure measurements are performed offline due to the solidification of the solution during the process. Therefore, to implement osmotic pressure-based control strategies in continuous bioproduction, a sensor that can measure osmotic pressure at a higher measurement frequency (e.g., in real time or near real time without solidifying the solution) is desired.

[0154] Raman spectroscopy is widely used in bioprocessing and has previously been used in both upstream and downstream applications. Raman spectroscopy involves measuring inelastic light scattering by a sample that is excited with light (e.g., laser light), typically in the visible, near-infrared, or near-ultraviolet regions. The Raman scattered light is typically energy shifted relative to the excitation light, and the energy shift provides insight into the vibrational modes of the sample. By resolving the frequency of the Raman scattered light (e.g., by directing the scattered light through a monochromator to spatially separate the frequencies of the scattered light for detection by a detector), various sample attributes can be determined, including, but not limited to, the type and / or concentration of various components of the sample.

[0155] Raman scattering measurements can be performed quickly and are therefore well suited to be performed in any of the methods described herein. As described above, multiple process parameters can be extracted from the Raman spectrum using univariate or multivariate analytical techniques. These process parameters can be used for automated recipe progression and / or closed-loop control in any of the methods described above.

[0156] In this example, a Raman scattering sensor was used to obtain Raman scattering intensity measurements to quantify the osmolality of a drug-containing solution over a range of target osmolalities during a sequential formulation excipient addition step. To evaluate the effectiveness of using Raman scattering for osmolality-based control, each expected formulation component (e.g., mAb protein, excipient 1, excipient 2) of the drug-containing solution was evaluated for Raman activity at concentrations expected to be observed during normal processing. Test solutions of each formulation component were dosed in water at several concentration levels, along with excipient buffer and test solutions of the target drug substance. Each of these target solutions was then measured using a Raman spectrometer (obtained from MarcMetri, Seattle, WA) with an 180 μL flow cell. The spectrometer laser power was 450 mW, and exposure time was optimized to achieve a detector saturation level of 50–80%. The number of scans was selected to achieve sufficient spectral quality and the desired measurement time. Taking into account the control scheme described herein, various measurement conditions were selected to achieve a measurement time of 20 seconds or less.

[0157] The acquired Raman spectra were overlaid to identify formulation components that were visible within the measurement wavelength range. A target spectrum for the formulated drug product (e.g., drug substance) was constructed based on the identified Raman visible components. It was observed that the Raman scattering method can measure the following types of formulation components: stabilizing salts and tonicity modifiers (i.e., arginine), sugars, cryoprotectants, and solubility excipients (i.e., sucrose).

[0158] Other formulation components had low visibility in the measured Raman spectra. These included surfactants (i.e., PS80) and chelating agents (i.e., EDTA). Without being bound by theory, the reason for the low visibility of these components is due to their relatively low target concentrations in the drug-containing solution.

[0159] To develop a Raman model for osmotic pressure measurements that allows for the monitoring and automatic feedback control described herein, a calibration dataset of Raman spectra and offline reference measurements was obtained by creating a synthetic flow channel that mimicked unit operating conditions. The flow channel began with a concentrated buffer and protein solution, and excipient buffer solutions were added stepwise over time. During flow channel operation, the Raman spectra were measured at 25-3500 cm under conditions similar to those described above. -1 Off-line samples were also collected and tested for osmolality by conventional measurements using an osmometer.

[0160] Models were developed using the multivariate modeling software SIMCA® by first trimming the spectra to spectral regions previously identified during measurement of the test solutions. After spectral trimming, first derivative preprocessing, Savitzky-Golay smoothing, and standard normal variate (SNV) normalization were applied. The preprocessed spectra were matched to offline reference measurements to form PLS models of osmolality. Latent variables for each model were selected based on leave-one-out cross-validation and selection of the minimum mean squared error of calibration and cross-validation. Generally, models selected for use had low minimum mean squared errors of calibration and cross-validation and a low R 2 and Q 2 Values ​​were greater than 90%, indicating accurate estimation of new data. Furthermore, the selected model covers the range of osmolality values ​​likely to be encountered during continuous formulation operation, helping to ensure that the model does not have to predict osmolality values ​​by extrapolating outside the calibration range (e.g., osmolality values ​​range from 60 to 700 mOsm / kg H2O, with a target osmolality of 383 mOsm / kg H2O).

[0161] To control the continuous pharmaceutical formulation process, Raman spectra were acquired using a MarqMetrix Raman spectrometer as a sensor. The spectrometer contained a 180 μL flow cell connected to the MarqMetrix Raman suite. The Raman spectral information acquired by the sensor was sent to synTQ, a process analysis and technical data management tool, using an OPC communication protocol. This tool converted the Raman spectral information into an osmolality value using SIMCA® QP. This value was sent by the synTQ tool to control the system hardware and was input into a specific proportional-integral-derivative (PID) controller-based loop for process control.

[0162] In this example, Raman measurements were taken from the drug-containing solution immediately after excipient addition to continuously measure osmolality and drug concentration. When the Raman-determined osmolality was used for control in this step, a target osmolality setpoint was input to the controller, and the excipient pump flow rate was adjusted using a feedback loop employing PID-based control. During this step, model performance was assessed by periodically removing and analyzing offline samples and comparing the in-line and offline measurements. Model performance was evaluated by calculating the root mean square error of prediction (RMSEP). An RMSEP of less than 5 mOsm / kg HO was considered acceptable for Raman-based osmolality predictions. Nevertheless, an RMSEP of up to 20 mOsm / kg HO was considered acceptable based on typical historical process variation in formulations.

[0163] In many embodiments, the process control methods described herein include determining a ratio of measurements of specific attribute values ​​and adjusting the relative flow rates of excipient and / or pharmaceutical solutions based on the ratio of measurements. As noted above, control based on osmolality ratio was also tested in this example.

[0164] Typically, when excipients are added to a drug-containing solution as part of a pharmaceutical formulation, the excipient buffer is concentrated at a specific ratio relative to the final drug stream. Because this excipient ratio is fixed, the ratio of the osmolality of the excipient buffer to the excipient-added drug stream is also the same. The osmolality of the excipient buffer and the excipient-added drug stream was continuously determined from Raman measurements using a model based on chemical measurements of both solutions, and the ratio of the osmolality values ​​of the excipient buffer and the excipient-added drug stream was used to adjust the flow rate of the excipient pump.

[0165] Similar to the method described above for controlling based on attribute ratios such as protein concentration ratio, a buffer:drug adjustment factor was continuously calculated to adjust the excipient pump flow rate for process control. If the adjustment factor is less than 1, the excipient pump flow rate will decrease and the drug flow will approach the target osmolality value, whereas if the adjustment factor is greater than 1, the opposite is true.

[0166] Note that in some embodiments, if there is a significant contribution from the protein to the osmolality value of the post-excipient drug-containing solution, a baseline value for the protein osmolality can be subtracted from the post-excipient osmolality determined as described above, or a third measurement of osmolality can be obtained for the pre-excipient drug-containing solution, and this osmolality value can be sequentially subtracted based on the performance of previous unit operations. The osmolality value of the pre-excipient drug-containing solution can generally be obtained using any of the methods described herein, including performing Raman scattering measurements and applying a multivariate model to the obtained Raman spectral information to extract the osmolality value.

[0167] Raman scattering measurements can generally be used to obtain osmolality values ​​for any solution within a continuous biological production system. In this example, Raman scattering measurements are used to measure the osmolality after excipient addition, before excipient addition, and in the excipient buffer. However, it should be understood that Raman scattering measurements can also be used to obtain osmolality measurements for other process solutions, including, but not limited to, process solutions obtained after one or more filtration, dilution, mixing, separation, and / or polishing steps. Such solutions may include, for example, filtered solutions, elution solutions from chromatography columns, solutions obtained from mixing vessels, solutions obtained from centrifugation, and solutions obtained from process sampling at many different locations within the biological production system.

[0168] As mentioned above, osmolality is a typical critical quality attribute during formulation execution and can be used to detect deviations in the formulation. In this example, osmolality was measured using Raman scattering measurements. However, other types of measurements and corresponding sensors may be used to obtain osmolality measurements for use in the methods described herein. For example, infrared and / or ultraviolet absorbance or reflectance measurements can be used to obtain a spectrum of the solution, and a multivariate chemical measurement-based method can be applied to the spectrum to obtain an osmolality value. As another example, a conductivity sensor can be used to measure the conductivity of the solution, and the osmolality value of the solution can be determined from the measured conductivity.

[0169] In this example, process control was performed by measuring the osmolality of the excipient and the post-excipient solution to adjust based on the ratio of excipient flow rates. It should be noted that while osmolality is a useful attribute for feedback control, measurements of any other attribute described herein for the excipient and post-excipient solution can similarly be used for adjustment based on the ratio of excipient flow rates. That is, adjustment of excipient flow rates based on the ratio of attribute values ​​of the excipient and post-excipient solution is not limited to the osmolality attribute alone, but can be performed based on measurements of a wide variety of different attribute values ​​of the solutions, including, but not limited to, any of the attributes generally described herein.

[0170] Other embodiments Although the present disclosure describes specific implementation examples, these should not be construed as limiting the scope of the disclosure, but rather as descriptions of features in specific embodiments. Features described in the context of separate embodiments can also generally be implemented in combination in a single embodiment. Conversely, various features described in the context of a single embodiment can also be implemented in multiple embodiments separately or in any suitable subcombination. Furthermore, while multiple features may be present in a particular combination and initially claimed as such, one or more features from a claimed combination can generally be excluded from that combination, and a claimed combination may refer to a subcombination or a variation of a subcombination.

[0171] In addition to the embodiments explicitly disclosed herein, it will be understood that various modifications may be made to the above-described embodiments without departing from the spirit and scope of the present disclosure. Accordingly, other implementations are within the scope of the following claims.

Claims

1. receiving a flowing first solution containing a biopharmaceutical; directing the flowing first solution along a flow path including a dilution location where a dilution device is in fluid communication with the flow path; introducing a second solution into the flowing first solution at the dilution location to form a flowing third solution; measuring the biopharmaceutical in the flowing first solution at a location upstream from the dilution location; measuring the biopharmaceutical in the flowing third solution at a location downstream from the dilution location; determining a relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream locations; and adjusting a flow rate of at least one of the first solution and the second solution based on the relative relationship.

2. 10. The method of claim 1, wherein measuring the biopharmaceutical at the upstream location comprises obtaining a measurement of a parameter of the first solution.

3. The method of claim 2 , wherein the measured value of the parameter is the refractive index of the first solution.

4. The method of claim 2 , wherein the measured value of the parameter is the conductivity of the first solution.

5. The method of claim 2 , wherein the measured value of the parameter is the absorbance of the first solution.

6. The method of claim 2 , wherein the measured value of the parameter is the transmittance of the first solution.

7. The method of claim 2 , wherein the measured value of the parameter is the reflectance of the first solution.

8. 3. The method of claim 2, wherein the measured value of the parameter is the concentration of the biopharmaceutical in the first solution.

9. The method of claim 2 , wherein the measurement of the parameter is measured at a single wavelength.

10. 10. The method of claim 9, wherein the single wavelength is in the ultraviolet region of the spectrum.

11. The method of claim 9 , wherein the single wavelength is in the visible spectral region.

12. 10. The method of claim 9, wherein the single wavelength is in the infrared spectral region.

13. 10. The method of claim 1, wherein measuring the biopharmaceutical at the upstream location comprises obtaining multiple measurements of the first solution.

14. 14. The method of claim 13, wherein the plurality of measurements comprises spectral information of the first solution at a plurality of wavelengths.

15. 15. The method of claim 14, further comprising analyzing the spectral information to determine a value derived from the spectral information.

16. 16. The method of claim 15, wherein analyzing the spectral information comprises using a calibrated chemometric model to determine a value derived from the spectral information.

17. 16. The method of claim 15, wherein the value derived from the spectral information is the concentration of the biopharmaceutical in the first solution or a quantity related to the concentration of the biopharmaceutical in the first solution.

18. 15. The method of claim 14, wherein the spectral information comprises an infrared spectrum of the first solution.

19. 15. The method of claim 14, wherein the spectral information comprises an ultraviolet spectrum of the first solution.

20. 15. The method of claim 14, wherein the spectral information comprises a Raman scattering spectrum of the first solution.

21. 20. The method of claim 18, further comprising measuring attenuated total reflectance of incident infrared light to obtain the infrared spectrum.

22. 4. The method of claim 3, further comprising measuring the refractive index of the first solution by measuring attenuated total reflection of incident infrared light from the first solution.

23. 10. The method of claim 1, wherein measuring the biopharmaceutical at the downstream location comprises obtaining a measurement of a parameter of the third solution.

24. 24. The method of claim 23, wherein the measured values ​​of the parameters of the third solution include at least one member of the group consisting of: refractive index of the third solution, conductivity of the third solution, absorptivity of the third solution, transmittance of the third solution, reflectance of the third solution, and concentration of a biopharmaceutical in the third solution.

25. 24. The method of claim 23, wherein the measurement of the parameter of the third solution is measured at a single wavelength.

26. 26. The method of claim 25, wherein the single wavelength comprises a wavelength in the ultraviolet, visible, or infrared spectral region.

27. 10. The method of claim 1, wherein measuring the biopharmaceutical at the downstream location comprises obtaining multiple measurements of the third solution.

28. 28. The method of claim 27, wherein the plurality of measurements comprises spectral information of the third solution at a plurality of wavelengths.

29. 30. The method of claim 28, further comprising analyzing the spectral information to determine a value derived from the spectral information.

30. 30. The method of claim 29, wherein analyzing the spectral information comprises using a calibrated chemometric model to determine a value derived from the spectral information.

31. 30. The method of claim 29, wherein the value derived from the spectral information is the concentration of the biopharmaceutical in the third solution or a quantity related to the concentration of the biopharmaceutical in the third solution.

32. 30. The method of claim 28, wherein the spectral information includes at least one member of the group consisting of an infrared spectrum of the third solution, an ultraviolet spectrum of the third solution, and a Raman scattering spectrum of the third solution.

33. 33. The method of claim 32, further comprising measuring attenuated total reflectance of incident infrared light to obtain the infrared spectrum.

34. 25. The method of claim 24, further comprising measuring the refractive index of the third solution by measuring attenuated total reflection of incident infrared light from the third solution.

35. measuring the biopharmaceutical at the upstream location; obtaining at least one of measurements of a parameter of the first solution and spectral information of the first solution; measuring the biopharmaceutical at the downstream location includes obtaining at least one of measurements of a parameter of the third solution and spectral information of the third solution; 2. The method of claim 1, wherein at least one of the measured parameter of the first solution and the spectral information of the first solution is measured using a measurement technique that is different from at least one of the measured parameter of the third solution and the spectral information of the third solution.

36. 36. The method of claim 35, wherein at least one of the measured parameter of the first solution and the spectral information of the first solution includes a different type of information than at least one of the measured parameter of the third solution and the spectral information of the third solution.

37. 36. The method of claim 35, wherein the measured parameter of the first solution and / or the spectral information of the first solution, and the measured parameter of the third solution and / or the spectral information of the third solution are each independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, concentration of the biopharmaceutical, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

38. measuring the biopharmaceutical at the upstream location; obtaining at least one of measurements of a parameter of the first solution and spectral information of the first solution; measuring the biopharmaceutical at the downstream location includes obtaining at least one of measurements of a parameter of the third solution and spectral information of the third solution; 2. The method of claim 1, wherein at least one of the measured parameter of the first solution and the spectral information of the first solution, and at least one of the measured parameter of the third solution and the spectral information of the third solution are measured using a common measurement technique.

39. 39. The method of claim 38, wherein the measured parameter of the first solution and / or the spectral information of the first solution, and the measured parameter of the third solution and / or the spectral information of the third solution are each independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, concentration of the biopharmaceutical, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

40. The method of claim 1 , wherein the second solution does not contain the biopharmaceutical.

41. 10. The method of claim 1, wherein the flow rate of the flowing first solution along the flow path is less than 2 mL / min.

42. 42. The method of claim 41, wherein the flow rate is less than 1 mL / min.

43. determining a relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream locations, obtaining a first value that correlates to the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in the first solution at the upstream location; obtaining a second value that correlates to the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in the third solution at the downstream location; calculating a comparison quantity based on the first and second values; and adjusting a flow rate of at least one of the first solution and the second solution based on the comparison amount.

44. 44. The method of claim 43, wherein the comparison amount is a ratio of the first value to the second value.

45. 44. The method of claim 43, wherein the comparison amount is a mathematical function of the first and second values.

46. 10. The method of claim 1, further comprising adjusting a flow rate of at least one of the first solution and the second solution until the value of the relative relationship falls within a range of target values.

47. 44. The method of claim 43, further comprising adjusting a flow rate of at least one of the first solution and the second solution until the value of the comparison quantity falls within a range of target values.

48. The method of claim 1 , wherein the biopharmaceutical is a protein.

49. 49. The method of claim 48, wherein the protein is an antibody, an antibody fragment, or comprises a portion of an antibody.

50. 10. The method of claim 1, wherein the biopharmaceutical in the first solution is an active pharmaceutical ingredient and the third solution is a pharmaceutical product.

51. the biopharmaceutical is a first biopharmaceutical, the dilution location is a first dilution location, the flow path includes a second dilution location downstream from the first dilution location, and the method further comprises: introducing a fourth solution into the flowing third solution at a second dilution location downstream from the location where the first biopharmaceutical is measured in the flowing third solution to form a flowing fifth solution; measuring the second biopharmaceutical in the flowing fifth solution at locations upstream and downstream from the second dilution location; determining a relative relationship between the measurements of the second biopharmaceutical at the upstream and downstream locations or between values ​​derived from the measurements of the second biopharmaceutical; and adjusting at least one of the flow rate of the third solution and the flow rate of the fourth solution based on the relative relationship of the second biopharmaceutical.

52. 52. The method of claim 51, wherein measuring the second biopharmaceutical at a location downstream from the second dilution location comprises obtaining a measurement of a parameter of the fifth solution.

53. 53. The method of claim 52, wherein the measured values ​​of the parameters of the fifth solution include at least one member of the group consisting of: refractive index of the fifth solution, conductivity of the fifth solution, absorptivity of the fifth solution, transmittance of the fifth solution, reflectance of the fifth solution, and concentration of the second biopharmaceutical in the fifth solution.

54. 53. The method of claim 52, wherein the measurement of the parameter of the fifth solution is measured at a single wavelength.

55. 55. The method of claim 54, wherein the single wavelength comprises a wavelength in the ultraviolet, visible, or infrared spectral region.

56. 52. The method of claim 51 , wherein measuring the second biopharmaceutical at a location downstream from the second dilution location comprises obtaining a plurality of measurements of the fifth solution.

57. 57. The method of claim 56, wherein the plurality of measurements comprises spectral information of the fifth solution at a plurality of wavelengths.

58. 58. The method of claim 57, further comprising analyzing the spectral information of the fifth solution to determine a value derived from the spectral information.

59. 60. The method of claim 58, wherein analyzing the spectral information of the fifth solution comprises using a calibrated chemometric model to determine a value derived from the spectral information.

60. 59. The method of claim 58, wherein the value derived from the spectral information is the concentration of the second biopharmaceutical in the fifth solution or a quantity related to the concentration of the second biopharmaceutical in the fifth solution.

61. 58. The method of claim 57, wherein the spectral information of the fifth solution includes at least one member of the group consisting of an infrared spectrum of the fifth solution, an ultraviolet spectrum of the fifth solution, and a Raman scattering spectrum of the fifth solution.

62. 62. The method of claim 61, further comprising measuring attenuated total reflectance of incident infrared radiation to obtain an infrared spectrum of the fifth solution.

63. 54. The method of claim 53, further comprising measuring the refractive index of the fifth solution by measuring attenuated total reflection of incident infrared light from the fifth solution.

64. 10. The method of claim 1, wherein the biopharmaceutical is measured at locations upstream and downstream from the dilution location by measuring different types of spectral information corresponding to each of the first and third solutions.

65. 52. The method of claim 51 , wherein the measurements of the second biopharmaceutical, or values ​​derived from the measurements of the second biopharmaceutical at the upstream and downstream locations, are of different types.

66. 52. The method of claim 51, comprising obtaining the measurements of the second biopharmaceutical at the upstream and downstream locations, or values ​​derived from the measurements of the second biopharmaceutical, using different measurement techniques.

67. 52. The method of claim 51, wherein the fourth solution does not contain the second biopharmaceutical.

68. 68. The method of claim 67, wherein the fourth solution does not contain the first biopharmaceutical.

69. 10. The method of claim 1, wherein the flowing first solution is received from a purification unit of a bioproduction system.

70. 70. The method of claim 69, wherein the purification unit comprises a tangential flow filtration unit.

71. a flow tube including an inlet; a reservoir connected to the flow tube at a dilution position; at least one flow regulator connected between the reservoir and the dilution position and / or between the inlet and the dilution position; a first sensor disposed in the flow conduit at an upstream position between the inlet and the dilution location; a second sensor disposed downstream of the flow tube between the outlet and the dilution location; a controller connected to the first and second sensors and the flow regulator, the first sensor is configured to measure a biopharmaceutical in a flowing first solution entering the inlet; the reservoir is configured to introduce the second solution into the flow tube at the dilution location to form a flowing third solution; the second sensor is configured to measure the biopharmaceutical in the flowing third solution; The controller: determining a relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream locations; The system is configured to adjust at least one flow regulator to control a flow rate of at least one of the first solution and the second solution based on the relative relationship.

72. 72. The system of claim 71, wherein the first sensor is configured to obtain a measurement of a parameter of the first solution.

73. 73. The system of claim 72, wherein the first sensor is a refractive index sensor configured to measure the refractive index of the first solution.

74. 73. The system of claim 72, wherein the first sensor is a conductivity sensor configured to measure the conductivity of the first solution.

75. 73. The system of claim 72, wherein the first sensor is an absorptivity sensor configured to measure the absorptivity of the first solution.

76. 73. The system of claim 72, wherein the first sensor is a transmittance sensor configured to measure the transmittance of the first solution.

77. 73. The system of claim 72, wherein the first sensor is a reflectance sensor configured to measure the reflectance of the first solution.

78. 73. The system of claim 72, wherein the first sensor is a concentration sensor configured to measure the concentration of the biopharmaceutical in the first solution.

79. 73. The system of claim 72, wherein the first sensor is configured to measure the value of the parameter at a single wavelength.

80. 80. The system of claim 79, wherein the single wavelength is in the ultraviolet region of the spectrum.

81. 80. The system of claim 79, wherein the single wavelength is in the visible spectral region.

82. 80. The system of claim 79, wherein the single wavelength is in the infrared spectral region.

83. 72. The system of claim 71, wherein the first sensor is configured to obtain a plurality of measurements of the first solution.

84. 74. The system of claim 73, wherein the plurality of measurements comprises spectral information of the first solution at a plurality of wavelengths.

85. 75. The system of claim 74, wherein the controller is configured to analyze the spectral information to determine a value derived from the spectral information.

86. 86. The system of claim 85, wherein the controller is configured to analyze the spectral information by using a calibrated chemometric model to determine a value derived from the spectral information.

87. 86. The system of claim 85, wherein the value derived from the spectral information is the concentration of the biopharmaceutical in the first solution or a quantity related to the concentration of the biopharmaceutical in the first solution.

88. 85. The system of claim 84, wherein the first sensor is configured to obtain an infrared spectrum of the first solution.

89. 85. The system of claim 84, wherein the first sensor is configured to obtain an ultraviolet spectrum of the first solution.

90. 85. The system of claim 84, wherein the first sensor is configured to obtain a Raman scattering spectrum of the first solution.

91. 89. The system of claim 88, wherein the first sensor is configured to measure attenuated total reflectance of incident infrared light to obtain the infrared spectrum.

92. 74. The system of claim 73, wherein the first sensor is configured to measure the refractive index of the first solution by measuring attenuated total reflection of incident infrared light from the first solution.

93. 72. The system of claim 71, wherein the second sensor is configured to obtain a measurement of a parameter of the third solution.

94. 94. The system of claim 93, wherein the second sensor is configured to measure at least one member of the group consisting of the refractive index of the third solution, the conductivity of the third solution, the absorptivity of the third solution, the transmittance of the third solution, the reflectance of the third solution, and the concentration of a biopharmaceutical in the third solution.

95. 94. The system of claim 93, wherein the second sensor is configured to measure the value of the parameter of the third solution at a single wavelength.

96. 96. The system of claim 95, wherein the single wavelength comprises a wavelength in the ultraviolet, visible, or infrared spectral region.

97. 72. The system of claim 71, wherein measuring the second sensor is configured to obtain a plurality of measurements of the third solution.

98. 98. The system of claim 97, wherein the plurality of measurements comprises spectral information of the third solution at a plurality of wavelengths.

99. 99. The system of claim 98, wherein the controller is configured to analyze the spectral information to determine a value derived from the spectral information.

100. 100. The system of claim 99, wherein the controller is configured to analyze the spectral information by using a calibrated chemometric model to determine a value derived from the spectral information.

101. 100. The system of claim 99, wherein the value derived from the spectral information is the concentration of the biopharmaceutical in the third solution or a quantity related to the concentration of the biopharmaceutical in the third solution.

102. 99. The system of claim 98, wherein the second sensor is configured to acquire spectral information including at least one member of the group consisting of an infrared spectrum of the third solution, an ultraviolet spectrum of the third solution, and a Raman scattering spectrum of the third solution.

103. 103. The system of claim 102, wherein the second sensor is configured to measure attenuated total reflection of incident infrared light to obtain the infrared spectrum.

104. 95. The system of claim 94, wherein the second sensor is configured to measure the refractive index of the third solution by measuring attenuated total reflection of incident infrared light from the third solution.

105. 72. The system of claim 71, wherein the first and second sensors are configured to measure the biopharmaceutical in the first and third solutions using different measurement techniques.

106. 106. The system of claim 105, wherein at least one of the measured parameter values ​​of the first solution and the spectral information of the first solution includes a different type of information than at least one of the measured parameter values ​​of the third solution and the spectral information of the third solution.

107. 106. The system of claim 105, wherein the measured value of a parameter of the first solution and / or the spectral information of the first solution, and the measured value of a parameter of the third solution and / or the spectral information of the third solution are each independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, concentration of the biopharmaceutical, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

108. 72. The system of claim 71, wherein the first and second sensors are configured to measure the biopharmaceutical in the first and third solutions using a common measurement technique.

109. 109. The system of claim 108, wherein the measured value of a parameter of the first solution and / or the spectral information of the first solution, and the measured value of a parameter of the third solution and / or the spectral information of the third solution are each independently selected from the group consisting of refractive index, conductivity, absorbance, transmittance, reflectance, concentration of the biopharmaceutical, infrared spectrum, ultraviolet spectrum, and Raman scattering spectrum.

110. 72. The system of claim 71, wherein the second solution does not contain the biopharmaceutical.

111. 72. The system of claim 71, wherein the first and second sensors are of different types.

112. The controller determines the relative relationship between the biopharmaceutical measurements or values ​​derived from the biopharmaceutical measurements at the upstream and downstream locations by: obtaining a first value that correlates to the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in the first solution; obtaining a second value that correlates to the concentration of the biopharmaceutical or an amount related to the concentration of the biopharmaceutical in the third solution; calculating a comparison quantity based on the first and second values; 72. The system of claim 71, configured to determine by adjusting at least one regulator to control a flow rate of at least one of the first solution and the second solution based on the comparison amount.

113. 113. The system of claim 112, wherein the controller is configured to calculate the comparison amount as a ratio of the first value and the second value.

114. 113. The system of claim 112, wherein the controller is configured to calculate the comparison amount as a mathematical function of the first and second values.

115. 72. The system of claim 71, wherein the controller is configured to adjust the at least one regulator to control a flow rate of at least one of the first solution and the second solution until the value of the relative relationship falls within a range of target values.

116. 113. The system of claim 112, wherein the controller is configured to adjust the at least one flow regulator to control the flow rate of at least one of the first solution and the second solution until the value of the comparison amount falls within a target value range.

117. 72. The system of claim 71, wherein the biopharmaceutical is a protein.

118. 118. The system of claim 117, wherein the protein is an antibody, an antibody fragment, or comprises a portion of an antibody.

119. 72. The system of claim 71, wherein the biopharmaceutical in the first solution is an active pharmaceutical ingredient and the third solution is a pharmaceutical product.

120. 72. The system of claim 71, further comprising a purification unit for use in a bioproduction system, the purification unit being in fluid communication with the inlet.

121. 121. The system of claim 120, wherein the purification unit comprises a tangential flow filtration unit.

122. 3. The method of claim 2, wherein the measured value of the parameter of the first solution is the osmolality of the first solution.

123. 24. The method of claim 23, wherein the measured value of the parameter of the third solution is the osmolality of the third solution.

124. 2. The method of claim 1, wherein measuring the biopharmaceutical at the upstream location comprises obtaining an osmolality value of the first solution, and measuring the biopharmaceutical at the downstream location comprises obtaining an osmolality value of the third solution.

125. 125. The method of claim 124, comprising adjusting the flow rate of the second solution based on the relative relationship.

126. 125. The method of claim 124, wherein the relative relationship is the ratio of the osmolality values ​​of the first and third solutions.

127. receiving a flowing first solution containing a biopharmaceutical; directing the flowing first solution along a flow path including a dilution location where a dilution device is in fluid communication with the flow path; introducing a second solution into the flowing first solution at the dilution location to form a flowing third solution; measuring a value of an attribute of the second solution at a location upstream from the dilution location; measuring an attribute value of the third solution at a location downstream from the dilution location; determining a relative relationship between the measured attribute values ​​of the second and third solutions; and adjusting the flow rate of the second solution based on the relative relationship.

128. 128. The method of claim 127, wherein the measured value of the attribute of the second solution is the osmolality of the second solution.

129. 128. The method of claim 127, wherein the measured value of the attribute of the second solution is the refractive index of the second solution.

130. 128. The method of claim 127, wherein the measured value of the attribute of the second solution is the conductivity of the second solution.

131. 128. The method of claim 127, wherein the measurement of the attribute of the second solution is the absorbance of the second solution.

132. 128. The method of claim 127, wherein the measurement of the attribute of the second solution is the transmittance of the second solution.

133. 128. The method of claim 127, wherein the measurement of the attribute of the second solution is the reflectance of the second solution.

134. 128. The method of claim 127, wherein the measurement of the attribute of the second solution is measured at a single wavelength.

135. 128. The method of claim 127, wherein measuring the value of the attribute of the second solution comprises obtaining a plurality of measurements of the second solution.

136. 136. The method of claim 135, wherein the plurality of measurements comprises spectral information of the second solution at a plurality of wavelengths.

137. 137. The method of claim 136, further comprising analyzing the spectral information to determine a value of the attribute of the second solution.

138. 138. The method of claim 137, wherein analyzing the spectral information comprises using a calibrated chemometric model to determine a value of the attribute of the second solution.

139. 139. The method of claim 138, wherein the value of the attribute is the osmolality of the second solution.

140. 137. The method of claim 136, wherein the spectral information comprises a Raman scattering spectrum of the second solution.

141. 137. The method of claim 136, wherein the spectral information comprises an infrared spectrum of the second solution.

142. 137. The method of claim 136, wherein the spectral information comprises an ultraviolet spectrum of the second solution.

143. 128. The method of claim 127, wherein the measured value of the attribute of the third solution is the osmolality of the third solution.

144. 128. The method of claim 127, wherein the measured value of the attribute of the second solution is the osmolality of the second solution and the measured value of the attribute of the third solution is the osmolality of the third solution.

145. 128. The method of claim 127, wherein the measured value of the attribute of the third solution comprises at least one member of the group consisting of a refractive index of the third solution, a conductivity of the third solution, an absorptivity of the third solution, a transmittance of the third solution, and a reflectance of the third solution.

146. 128. The method of claim 127, wherein the measurement of the attribute of the third solution is measured at a single wavelength.

147. 128. The method of claim 127, wherein measuring the value of the attribute of the third solution comprises obtaining a plurality of measurements of the third solution.

148. 148. The method of claim 147, wherein the plurality of measurements comprises spectral information of the third solution at a plurality of wavelengths.

149. 149. The method of claim 148, further comprising analyzing the spectral information to determine a value of the attribute of the third solution.

150. 150. The method of claim 149, wherein analyzing the spectral information includes using a calibrated chemometric model to determine a value of the attribute of the third solution.

151. 151. The method of claim 150, wherein the value of the attribute is the osmolality of the third solution.

152. 149. The method of claim 148, wherein the spectral information comprises a Raman scattering spectrum of the third solution.

153. 149. The method of claim 148, wherein the spectral information comprises an infrared spectrum of the third solution.

154. 149. The method of claim 148, wherein the spectral information comprises an ultraviolet spectrum of the third solution.

155. 128. The method of claim 127, wherein the values ​​of the attribute of the second and third solutions are measured using different measurement techniques.

156. 156. The method of claim 155, wherein the attributes of the second and third solutions for which values ​​are measured are different.

157. 128. The method of claim 127, wherein the second solution does not contain the biopharmaceutical.

158. 128. The method of claim 127, wherein the flow rate of the flowing first solution along the flow path is less than 2 mL / min.

159. 128. The method of claim 127, wherein determining the relative relationship between the measured attribute values ​​of the second and third solutions comprises calculating a comparative quantity between the measured attribute values.

160. 160. The method of claim 159, wherein the comparison quantity is a ratio of the measured attribute values.

161. 160. The method of claim 159, wherein the comparison quantity is a mathematical function of the measured attribute value.

162. 128. The method of claim 127, further comprising adjusting the flow rate of the second solution until the value of the relative relationship falls within a range of target values.

163. 160. The method of claim 159, further comprising adjusting the flow rate of the second solution until the value of the comparison amount falls within a target value range.

164. 128. The method of claim 127, wherein the biopharmaceutical is a protein.

165. 165. The method of claim 164, wherein the protein is an antibody, an antibody fragment, or comprises a portion of an antibody.

166. 128. The method of claim 127, wherein the biopharmaceutical in the first solution is an active pharmaceutical ingredient and the third solution is a pharmaceutical product.

167. a flow tube including an inlet; a reservoir connected to the flow tube at a dilution position; at least one flow regulator connected between the reservoir and the dilution location; a first sensor disposed between the reservoir and the dilution position; a second sensor disposed downstream between the outlet of the flow tube and the dilution location; a controller connected to the first and second sensors and the flow regulator, the flow tube is configured to receive a flowing first solution containing a biopharmaceutical through the inlet; the reservoir is configured to introduce a second solution into the flow tube at the dilution location to form a flowing third solution; the first sensor is configured to measure a value of an attribute of the second solution; the second sensor is configured to measure a value of an attribute of the third solution; The controller: determining a relative relationship between the measured attribute values ​​of the second and third solutions; The system is configured to adjust at least one flow regulator to control the flow rate of the second solution based on the relative relationship.

168. 168. The system of claim 167, wherein the first sensor is a Raman scattering sensor configured to measure Raman scattered light from the second solution.

169. 168. The system of claim 167, wherein the first sensor is a refractive index sensor configured to measure the refractive index of the second solution.

170. 168. The system of claim 167, wherein the first sensor is a conductivity sensor configured to measure the conductivity of the second solution.

171. 168. The system of claim 167, wherein the first sensor is an absorbance sensor configured to measure the absorbance of the second solution.

172. 168. The system of claim 167, wherein the first sensor is a transmittance sensor configured to measure the transmittance of the second solution.

173. 168. The system of claim 167, wherein the first sensor is a reflectance sensor configured to measure the reflectance of the second solution.

174. 168. The system of claim 167, wherein the first sensor is configured to obtain multiple measurements of the second solution.

175. 175. The system of claim 174, wherein the plurality of measurements comprises spectral information of the second solution at a plurality of wavelengths.

176. 176. The system of claim 175, wherein the controller is configured to analyze the spectral information to determine the measured attribute value of the second solution.

177. 177. The system of claim 176, wherein the controller is configured to analyze the spectral information by using a calibrated chemometric model to determine the measured attribute value of the second solution from the spectral information.

178. 176. The system of claim 175, wherein the first sensor is configured to acquire a Raman scattering spectrum of the second solution.

179. 168. The system of claim 167, wherein the second sensor is a Raman scattering sensor configured to measure Raman scattered light from the third solution.

180. 168. The system of claim 167, wherein the second sensor comprises at least one element of the group consisting of a refractive index sensor configured to measure the refractive index of the third solution, a conductivity sensor configured to measure the conductivity of the third solution, an absorptivity sensor configured to measure the absorptivity of the third solution, a transmittance sensor configured to measure the transmittance of the third solution, and a reflectance sensor configured to measure the reflectance of the third solution.

181. 168. The system of claim 167, wherein the second sensor is configured to obtain multiple measurements of the third solution.

182. 182. The system of claim 181, wherein the plurality of measurements includes spectral information of the third solution at a plurality of wavelengths.

183. 183. The system of claim 182, wherein the controller is configured to analyze the spectral information to determine the measured attribute value of the third solution from the spectral information.

184. 184. The system of claim 183, wherein the controller is configured to analyze the spectral information by using a calibrated chemometric model to determine the measured attribute value of the third solution.

185. 183. The system of claim 182, wherein the second sensor is configured to acquire a Raman scattering spectrum of the third solution.

186. 168. The system of claim 167, wherein the first and second sensors are configured to measure attribute values ​​of the second and third solutions using different measurement techniques.

187. 187. The system of claim 186, wherein the attributes of the second and third solutions for which the values ​​are measured are different.

188. 168. The system of claim 167, wherein the second solution does not contain the biopharmaceutical.

189. 68. The system of claim 67, wherein the first and second sensors are of different types.

190. The controller: calculating a comparison quantity based on the measured attribute values ​​of the second and third solutions; 68. The system of claim 67, configured to adjust the at least one regulator to control a flow rate of the second solution based on the comparison amount.

191. 191. The system of claim 190, wherein the controller is configured to calculate the comparison amount as a ratio of the measured attribute values.

192. 191. The system of claim 190, wherein the controller is configured to calculate the comparison quantity as a mathematical function of the measured attribute value.

193. 168. The system of claim 167, wherein the controller is configured to adjust the at least one regulator to control the flow rate of the second solution until the value of the relative relationship falls within a target value range.

194. 191. The system of claim 190, wherein the controller is configured to adjust the at least one flow regulator to control the flow rate of the second solution until the value of the comparison amount falls within a target value range.

195. The system of claim 167, wherein the biopharmaceutical is a protein.

196. 196. The system of claim 195, wherein the protein is an antibody, an antibody fragment, or comprises a portion of an antibody.

197. 168. The system of claim 167, wherein the biopharmaceutical in the first solution is an active pharmaceutical ingredient and the third solution is a pharmaceutical product.

198. 168. The system of claim 167, wherein the purification unit further comprises a purification unit in fluid communication with the inlet.

199. 200. The system of claim 198, wherein the purification unit comprises a tangential flow filtration unit.

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