Membrane Chromatography Systems

Abstract

In summary, the present invention provides a chromatography system configured to process a feed fluid containing a plurality of components, where at least one of the components of the feed fluid is a target component. The chromatography system includes a flow path including a plurality of fluid control components configured to control fluid flow, and a stationary phase, the stationary phase being at least one membrane adsorber connected to the flow path, the stationary phase configured to isolate the target component. The flow path is configured to optimize collection of the target component.

Description

[Technical Field]

[0001] The following description relates to a system for membrane chromatography. [Background technology]

[0002] Biopharmaceutical or pharmaceutical manufacturing involves the purification of solutions from which active pharmaceutical ingredients (APIs) are extracted. These solutions, also known as feeds, can be produced chemically or bioorganically. The feed contains multiple components that need to be separated from each other, such as one or more target components and impurities. Chromatography is the technique used to perform this separation process.

[0003] The details of the chromatographic separation process can be selected from multiple available options, for example, regarding the interaction mechanism, process type, and stationary phase. Based on the interaction mechanism, chromatography can be classified as, for example, ion exchange, hydrophobic interaction, affinity, or mixed-mode chromatography. The process type can relate to the time aspect of the process execution, such as batch or continuous operation, and / or the aspect of the interaction between the target product and the separation medium in the stationary phase, such as capture, binding, and elution or flow-through. Chromatographic stationary phases can be, for example, particle / resin-based, membrane-based, monolith-based, and fiber-based.

[0004] The greatest difference in chromatographic process steps arises from kinetically confined and non-kinetically confined stationary phases. Dynamically confined stationary phases are typically stationary phases, such as resin-based stationary phases, that require a residence time of greater than or about 2 minutes to bind, while non-kinetically confined stationary phases are stationary phases, such as membrane-based stationary phases, that require less than 2 minutes to bind.

[0005] A chromatography process typically consists of multiple cycles, each of which includes equilibration, loading, one or more wash steps, elution, regeneration, and cleaning-in-place (CIP). During equilibration, the stationary phase is prepared for the load step, during which the binding sites of each target substance must be freely accessible. The equilibration buffer removes residues from previous steps, such as CIP, that could adversely affect the binding properties of the stationary phase.

[0006] During loading, a feed containing a mixture of components is forced to interact with a stationary phase, where one or more components remain on the stationary phase while others migrate. For example, in bind-and-elute mode, the target components must bind to the stationary phase, and the components to be separated must pass through the stationary phase. Unbound components are rinsed away in a wash step to prevent carryover with the target components in the eluate during elution.

[0007] During elution, the target component is displaced from the binding sites of the stationary phase by the elution buffer due to changes in the physicochemical environment (pH value, conductivity) of the stationary phase, and then recovered. During regeneration, changes in the physicochemical properties of the environment on the stationary phase, which are more pronounced than those in the elution step, dissolve poorly soluble impurities (e.g., lipids, dyes, DNA) and restore the binding capacity of the stationary phase. A clean-in-place (CIP) step is performed to reduce bioburden and eliminate contaminants that may remain on the stationary phase even after the regeneration step. The CIP step is generally performed using a caustic soda solution.

[0008] The traditional chromatographic technique is column chromatography, where the stationary phase (also called the "separation medium") is a resin placed in a column or tube. The separation mechanism is determined by the physicochemical properties of the resin.

[0009] Two parameters that can be adjusted for column design are the column diameter and the packing height of the separation medium. These determine the volume of the stationary phase, which should be selected based on the volume of the feed being processed, as well as the concentration of the target components present therein and the binding capacity of the separation medium. Depending on influencing factors such as the cost of the separation medium relative to the process time and / or the cost of buffer consumption, the stationary phase volume is selected so that 2 to 20 cycles are performed to process the entire feed volume.

[0010] In particular, the packing height of the separation medium should be selected such that: - any non-uniformity in the column bed and the associated loss of separation efficiency, productivity and resolution are compensated for over the distance covered by the feed during its passage through the column; - the residence time of the feed on the stationary phase required to achieve the highest possible binding capacity is achieved; - The pressure drop / back pressure of the packed separation medium does not exceed a threshold for the specified residence time and resulting flow rate. For example, if a chromatography column is packed with a packing pressure of 3 bar, this pressure must not be exceeded during the process, otherwise the packed bed will compress and a headspace (i.e., a liquid-filled cavity between the top of the column and the bed of separation medium compressed by the overpressure) will form within the column, which will impair the process and the separation mechanism.

[0011] The column diameter should be selected as follows: - Depending on the filling height, the amount of volume of the filled separation medium is such that the process time is as short as possible, and the binding capacity is directly proportional to the volume of the filled separation medium; The cost for the required volume of separation medium does not exceed a given threshold, depending on the process time and number of cycles.

[0012] The duration of a column chromatography cycle is usually more than 10 minutes and often in the range of several hours. For example, in a Protein A column chromatography where an antibody-containing feed is processed, with a packing height of 20 cm, a column diameter of 25 cm, and a column volume of 9.8 L, the cycle takes approximately 4 hours, as shown in more detail in the table below: [Table 1]

[0013] The volume of each step, expressed in units of column volume, refers to the volume of a given fluid / media required for a given step.

[0014] An alternative to column chromatography is membrane chromatography, in which the stationary phase is provided by one or more membrane adsorber, i.e., microporous or macroporous membranes derivatized with functional groups similar to those on the resin. Exemplary membrane adsorber are disclosed in European Patent Application Publication No. EP 2 274 081 A1.

[0015] The residence time on the membrane adsorber is shorter than on the resin because convection is primarily responsible for mass transport, resulting in more efficient adsorption relative to the diffusion mechanism that predominates on the resin. Therefore, the productivity of the process, i.e., the amount of product obtained per unit of feed volume and per unit of time (usually expressed as g / (L × hr)), can be increased by a factor of more than three times over resin-based column chromatography. Furthermore, in contrast to column chromatography, the stationary phase is almost fully utilized in membrane chromatography. Summary of the Invention [Problem to be solved by the invention]

[0016] It is an object of the present invention to provide a membrane chromatography system with improved process quality and efficiency, for example, as quantified by one or more of the following factors: the amount of target component(s) obtained, the purity of the target component(s) obtained, the amount of time required to perform the process, and the amount of resources (such as buffers) required to perform the process. [Means for solving the problem]

[0017] The achievement of this object according to the invention is set forth in the independent claims. Further developments of the invention are the subject of the dependent claims.

[0018] According to one aspect, there is provided a chromatography system configured to process a feed fluid containing a plurality of components, wherein at least one of the plurality of components of the feed fluid is a target component. a flow path including a plurality of fluid control components configured to control fluid flow; a stationary phase, wherein the stationary phase is at least one membrane adsorber connected to the flow path, and the stationary phase is configured to isolate a target component; Equipped with Here, the flow path is configured to optimize collection of the target component.

[0019] A feed fluid (also referred to as a "feed") contains multiple components or substances, at least one of which is a component of interest, the target component. The goal of a chromatographic process is to harvest the target component, i.e., to separate it from other components of the feed, and then recover the target component as a product of the process.

[0020] The product is made substantially from the target component, which means that the degree of purity, i.e., the relative amount of the target component in the total product (e.g., by weight, mass, volume), approaches 100%, e.g., greater than 99%. Product purity is particularly important, for example, for pharmaceutical and biopharmaceutical applications.

[0021] The feed may contain more than one target component that is collected so that the chromatographic process may yield more than one product. Additionally, intermediates and / or by-products may also be obtained and recovered when performing the chromatographic process. In some cases, different concentrations of the target component may be recovered separately.

[0022] Components of the feed fluid that are not of interest (also referred to as "scrap components"), as well as other materials used during the chromatographic process, such as buffers, form waste products of the process and are recovered together.

[0023] By way of example, the feed fluid may be a solution, such as a protein-containing solution or a cell-containing solution. Examples of cell-containing solutions include vaccines or other solutions containing viruses, as well as solutions containing mammalian cells. Examples of protein-containing solutions include liquids containing therapeutic proteins (e.g., monoclonal antibodies, enzymes, hormones, etc.). In these cases, the target component may be a solute, such as a specific type of cell or a specific protein. Scrap components may include DNA, salts, and host cell proteins (HCPs).

[0024] A chromatography system includes a fluid path, which comprises means for permitting fluid flow, such as, for example, conduits and vessels for receiving, transporting, and / or containing fluids, and means for regulating fluid flow, such as valves, pumps, sensors, and filters. Thus, the fluid path comprises a number of fluid control components configured, among other things, to control fluid flow.

[0025] A flow path connects one or more entry points, where one or more fluids are introduced into the flow path, to one or more exit points, where one or more fluids are released from the flow path. A flow path may include different alternative paths connecting an entry point to an exit point.

[0026] The direction from the entry point(s) to the exit point(s) is the forward direction of the flow path, which is the general macroscopic direction of fluid flow. The phrase "X is located after Y" indicates that X is after Y in the forward direction of the fluid path, i.e., X is closer to the exit point(s) than Y and farther from the entry point(s) than Y. Similarly, "X is located before Y" indicates that X is before Y in the forward direction of the fluid path, i.e., X is closer to the entry point(s) than Y and farther from the exit point(s) than Y.

[0027] Fluids flowing through the flow path include one or more buffers, wash solutions, feed fluids (upstream of the stationary phase), and separated components of the feed fluid (downstream of the stationary phase). A buffer (or "buffer solution") is an aqueous solution that effectively resists and prevents large changes in pH following the addition of an acid or base. This is due to the presence of either a weak acid and its conjugate base or a weak base and its conjugate acid. Chromatography processes may require different buffers at different steps in the cycle.

[0028] A chromatography system includes a stationary phase. The stationary phase is the part of the chromatography system configured to separate the target component(s) from scrap components that are initially mixed together in a feed fluid. The feed fluid is optionally mixed with other substances (e.g., water for dilution), and a buffer represents the mobile phase. In particular, the stationary phase can isolate and then recover the target component(s) in the mobile phase.

[0029] Specifically, the stationary phase is one or more membrane adsorbers. In the case of multiple membrane adsorbers, they may be stacked on top of each other and / or arranged in series or parallel on two or more alternative paths of the flow path. The membrane adsorber(s) may adsorb a target component that is subsequently recovered, for example, by elution. Alternatively, the target component may flow through, i.e., not be adsorbed by the membrane adsorber, while other components of the feed fluid are adsorbed.

[0030] The membrane adsorber(s) may be made of, for example, stabilized reinforced cellulose with sulfonic acid or salt-tolerant anion exchanger or phenyl as the ligand. Other materials may be used for the membrane adsorber, and other substances may be used as the ligand. The membrane adsorber(s) may be provided in a capsule or frame, for example, made of plastic.

[0031] At least one membrane adsorber is connected to a flow path. When connected to a flow path, the membrane adsorber becomes part of the flow path in the sense that a fluid can flow through the membrane adsorber. Illustratively, the membrane adsorber(s) may be connected to the flow path by a valve called a "membrane valve." For example, a membrane adsorber (or a stack of membrane adsorbers) may be connected to the flow path by a pair of membrane valves, one placed before the membrane adsorber and one placed after the membrane adsorber. In this case, the flow path may include a path that passes through the membrane adsorber and an alternative path that does not pass through the membrane adsorber. Alternatively, the membrane adsorber may be connected directly (i.e., without a valve) to a pipe or other conveying means of the flow path, and there may be only one path that passes through the membrane adsorber.

[0032] The use of membrane adsorber as the stationary phase in a chromatography system allows for a larger operating flow rate range compared to column chromatography with an equivalent volume of stationary phase. Illustratively, the operating flow rate range for resins may be 0.05-2 CV / min, more preferably 0.1-1 CV / min, and most preferably 0.1-0.5 CV / min, while the operating flow rate range for membrane adsorber may be 0.5-40 MV / min, more preferably 1-30 MV / min, and most preferably 3-20 MV / min.

[0033] The flow path of the chromatography system is configured to optimize the collection of target components by the membrane adsorber(s). The flow path, and in particular its fluid control components, are adapted to the operation of the membrane adsorber(s). In other words, the dynamics of fluid control within the flow path are adapted to the highly dynamic characteristics / behavior of the membrane adsorber(s). In particular, the compositional / structural design of the flow path and / or the control of fluid flow provided by the flow path are optimally configured for membrane chromatography.

[0034] The chromatography system may include or be configured to be connected to a control system including a processor configured to manage fluid control components, such as, for example, a distributed control system, which may receive signals from one or more fluid control components, such as sensors, and may send signals to one or more fluid control components, such as valves and pumps.

[0035] In a particular example, the plurality of fluid control components includes: a first outlet valve configured to be connected to the at least one membrane adsorber and to a target component collection vessel; a second outlet valve connected to the at least one membrane adsorber and configured to be connected to a waste collection vessel; and Here, the first outlet valve and the second outlet valve have a switching time of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds.

[0036] The first and second outlet valves are arranged after the at least one membrane adsorber. In particular, the first and second outlet valves may be arranged at the ends of the flow paths.

[0037] The first and second outlet valves are fluidly connected to the at least one membrane adsorber in the sense that there is a fluid path from the at least one membrane adsorber to each of the first and second outlet valves. In other words, fluid can flow from the at least one membrane adsorber toward the outlet valve. For example, one or more pipes can connect the at least one membrane adsorber to the outlet valve.

[0038] In some examples, one or more other fluid control components may be interposed between the at least one membrane adsorber and the outlet valve, such as a sensor and / or another valve (eg, a membrane valve).

[0039] One of the outlet valves, for example the first outlet valve, is configured to be connected, for example by tubing, to a target component collection container. In other words, the first outlet valve is dedicated to the discharge of the target component from the flow path to an external container. The second outlet valve is configured to be connected, for example by tubing, to a waste collection container. In other words, the second outlet valve is dedicated to the discharge of waste (e.g., used buffer, DNA, ...) from the flow path to an external container.

[0040] Thus, depending on the fluid entering the outlet valve from the flow path, in particular from at least one membrane adsorber, one outlet valve is open and the other is closed. Each valve can be switched between an open and a closed position according to a control signal, for example, from a control system. Illustratively, the control signal can be based on a sensor that determines the content of (a portion of) the fluid flowing towards the outlet valve at any given time, for example, an adsorption detector such as an ultraviolet (UV) sensor positioned between the membrane adsorber and the outlet valve. If the fluid contains target component molecules or particles above a certain predetermined threshold, the fluid can be referred to as a "target component fluid"; otherwise, the fluid can be referred to as a "waste fluid."

[0041] The switching time of a valve is the amount of time required for the valve to go from an open position to a closed position, or vice versa. Both the first outlet valve and the second outlet valve have a switching time of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds. In other words, the first outlet valve and the second outlet valve are controlled to switch from an open / closed position to an open / closed position in a time of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds.

[0042] This relatively short switching time has two advantages, given the high flow rates at which membrane adsorber devices operate. First, the fact that the first outlet valve can switch from closed to open in such a short time helps eliminate or reduce potential losses of target component. Indeed, if the target component fluid reaches a point in the flow path where the first outlet valve is located while the first outlet valve is still closed, some of the target component fluid may not be properly collected in the target component collection vessel.

[0043] Second, the fact that both outlet valves have short switching times further reduces the potential loss of target components and also reduces or eliminates the problem of backmixing, which refers to the unwanted mixing of fluids due to fluid movement in a direction other than the forward direction of the flow path.

[0044] Backmixing of fluids downstream of the membrane adsorber, especially at the outlet, can result in undesirable mixing of the target component fluid with other fluids. This can affect the purity of the recovered target component and can increase, for example, the elution volume. The elution volume is the volume of the fraction containing the target component from the start of elution to the end of elution, and therefore must be as low as possible to not only increase the concentration but also reduce the effort of subsequent processes, such as storage, process time, or buffer consumption.

[0045] The problem of backmixing is more relevant to membrane chromatography than to column chromatography. One reason is that the volume of the stationary phase is larger in column chromatography; for example, depending on the process design, the equivalent column volume for a 150 mL membrane volume is 10 L. As a result, the relative change in elution volume is negligible in column chromatography. For example, if a 150 mL membrane adsorber has an elution volume of 200 mL and an equivalent 10 L chromatography column has an elution volume of 15 L, then when 50 mL of backmixing fluid is added, the increase in the membrane chromatography system is 25% and the increase in the column chromatography system is 0.3%.

[0046] Furthermore, due to the smaller stationary phase volume and lower binding capacity per cycle compared to conventional column chromatography, a greater number of membrane chromatography cycles are required to process a given volume of feed compared to column chromatography. Given the relatively large number of cycles, the adverse effects of backmixing are substantially increased, rendering the membrane chromatography process ineffective.

[0047] A short switching time of the outlet valve (especially from open to closed) reduces the amount of fluid that can backflow through the valve and also reduces or eliminates the overlap period during which both valves may be open, thereby allowing for faster and more precise changes in the configuration of the flow path to match the dynamic characteristics of the membrane adsorber(s).

[0048] It should be noted that in column chromatography systems, longer valve switching times are selected (ie, about 3 seconds or longer) to protect the packed column bed from rapid pressure surges.

[0049] Another means for minimizing backmixing is to minimize the dead volume of the system, where the dead volume is the volume of the flow path (i.e., the system without the stationary phase) and is therefore given by the sum of the volumes of the conduits of the valve and all other elements through which the fluid flows from the inlet to the outlet. Thus, illustratively, the ratio of dead volume to stationary phase volume (not taking into account the porosity of the stationary phase) can be less than 5, preferably less than 4, more preferably less than 3, and most preferably less than 2. A smaller dead volume also helps to shorten the cycle duration.

[0050] In some examples, the fluid control component of the flow path may further comprise one or more additional outlet valves disposed after at least one membrane adsorber, each of which may be dedicated to the discharge of a different component, e.g., a by-product, and may be configured to be connected to an external container.

[0051] The additional outlet valve(s) may also have a switching time of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds.

[0052] In another example, the second outlet valve may have a switching time of about 3 seconds or more, and the plurality of fluid control components may further include a check valve disposed after the second outlet valve. A check valve is a valve that allows fluid to flow in only one direction. In particular, a check valve only allows fluid, such as waste liquid, to flow toward a waste collection container, not in the reverse direction. The presence of the check valve reduces or eliminates the problem of backmixing.

[0053] In this example, the first outlet valve may have a switching time of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds. Alternatively, the first outlet valve may have a switching time of about 3 seconds or greater than about 3 seconds. In this case, optionally, the plurality of fluid control components may further comprise another check valve disposed after the first outlet valve.

[0054] In a particular example, the plurality of fluid control components comprises: a first inlet valve configured to be connected to a supply fluid source; a second inlet valve configured to be connected to a buffer source; may further comprise: Here, the first inlet valve and the second inlet valve have a switching time of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds.

[0055] The first and second inlet valves are arranged before the at least one membrane adsorber. In particular, the first and second outlet valves may be arranged at the beginning of the flow path.

[0056] The first and second outlet valves are fluidly connected to the at least one membrane adsorber in the sense that there is a fluid path from each of the first and second outlet valves to the at least one membrane adsorber. In other words, fluid can flow from the inlet valve toward the at least one membrane adsorber. For example, one or more pipes can connect the at least one membrane adsorber to the inlet valve.

[0057] In some examples, one or more other fluid control components may be interposed between the at least one membrane adsorber and the inlet valve, such as a pump, a filter, a sensor (e.g., a UV sensor), and / or another valve (e.g., a membrane valve).

[0058] The first inlet valve is configured to be connected to a source of feed fluid, for example by tubing. In other words, the first inlet valve is dedicated to input of a feed from an external source to the flow path. The second inlet valve is configured to be connected to a buffer source, for example by tubing. In other words, the second inlet valve is dedicated to input of a buffer from an external source to the flow path. The second inlet valve may be connected to multiple buffer sources, each providing a different buffer. Alternatively, the flow path may comprise multiple (second) inlet valves configured to be connected to multiple buffer sources, respectively.

[0059] In one example, the flow path can include two inlet lines, one beginning with a first inlet valve and the other beginning with a second inlet valve, where the two inlet lines represent two parallel branches of the flow path that meet before at least one membrane adsorber. In the case of multiple second inlet valves, the flow path can include three or more inlet lines. In another example, the flow path can include a single inlet line, at the beginning of which the first and second inlet valves are located.

[0060] Illustratively, the flow path may include at least one pump. Each inlet line may have a pump for driving the respective fluid flow at a defined flow rate. The pump may be specifically integrated into the chromatography system, i.e., may be a fixed element of the flow path, to avoid additional dead volumes due to connecting elements.

[0061] Backmixing is not only a problem downstream of the membrane adsorber(s). Backmixing of fluids before and / or above the membrane adsorber(s) results in changes in fluid properties, reducing the binding capacity and / or elution profile of the target components due to the small membrane volume. As previously described, upstream backmixing also has less adverse effect on the separation properties / binding capacity of the column compared to membrane adsorber(s).

[0062] The short switching time of the inlet valves reduces or eliminates backmixing problems upstream of the membrane adsorber(s). In another example, the first and second inlet valves may have switching times of about 3 seconds or greater than about 3 seconds, and the plurality of fluid control components may further comprise at least one inlet check valve disposed after the first and second inlet valves. Illustratively, when the flow path includes at least one pump, the at least one inlet check valve may be disposed after the at least one pump.

[0063] In instances where there are multiple inlet lines, there may be multiple corresponding check valves. In instances where there is only one inlet line, there may be only one inlet check valve.

[0064] More generally, all valves in a chromatography system may have switching times less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds, or alternatively, all valves in a chromatography system may be followed by check valves.

[0065] In the case of multiple inlet lines, the flow path may further comprise a mixing valve in each inlet line, for example a first mixing valve and a second mixing valve. The mixing valves may prevent back-mixing between the different input fluids.

[0066] In certain examples, the plurality of fluid control components may further comprise a filter configured to filter only the feed fluid. The filter may be used, among other things, for particle separation and bioburden reduction to avoid blocking of the stationary phase. The filter may be positioned before at least one membrane adsorber in the flow path. The filter may be positioned after an inlet valve configured to admit the feed fluid into the flow path.

[0067] Illustratively, the flow path may comprise a dedicated inlet line for the feed fluid ("feed inlet line"), i.e., an inlet line used only for feeding. In other words, only the feed fluid flows through the feed inlet line. A filter may be placed directly in this feed inlet line (i.e., without a valve) so that only the feed fluid flows through the filter.

[0068] In other examples, the filter may be placed in part of a flow path through which a different fluid flows. In this case, the filter may be connected to a conduit by two valves ("filter valves") to create a path through which only the feed can flow, while the flow path provides an alternative path for other fluids.

[0069] The presence of a filter through which only the feed fluid flows helps reduce or eliminate backmixing.

[0070] In a particular example, the at least one membrane adsorber has a first pore size and the filter has a second pore size, the second pore size being smaller than the first pore size. Illustratively, the first pore size may be in the range of about 3 μm to about 5 μm, while the second pore size may be in the range of about 0.2 μm to about 0.8 μm.

[0071] In this way, only gas bubbles can reach the stationary phase where they are sufficiently dispersed to flow through it, avoiding a reduction in binding capacity. Therefore, bubble traps, as typically implemented in column chromatography systems, can be omitted, thus avoiding additional dead volume.

[0072] In certain examples, the plurality of fluid control components may further comprise an adsorption detector positioned after the at least one membrane adsorber, wherein the sampling rate of the adsorption detector is less than about 0.7 seconds, preferably less than about 0.5 seconds, and more preferably about 0.3 seconds or less.

[0073] As described above, an adsorption detector can be used to monitor the target component. For example, a UV sensor can record adsorption at a defined wavelength (e.g., 280 nm). The elution or recovery of the target component is performed by opening a dedicated outlet valve (first outlet valve) controlled by this UV signal. The sampling rate of the UV sensor corresponds to the time interval between subsequent recordings of measurement points and directly correlates with the product recovery rate. A sampling rate of less than about 0.7 seconds, preferably less than about 0.5 seconds, and more preferably less than about 0.3 seconds ensures that yield losses are reduced or eliminated even at high flow rates in membrane chromatography.

[0074] In certain examples, the plurality of fluid control components may further comprise an adsorption detector, such as a UV sensor, positioned before the at least one membrane adsorber. The presence of an adsorption detector upstream of the at least one membrane adsorber prevents system malfunction and / or damage and enables predictive and adaptive process control, as described below.

[0075] If the UV signal changes above / below a predefined threshold, the process can be stopped (e.g., by stopping the pump) and the cause (e.g., heterogeneity, microbial contamination, incorrectness of connected intermediates) can be eliminated. Thus, product loss or changes in the physicochemical properties of the stationary phase, for example due to incorrect buffers, do not occur. This can also be relevant for interconnected processes, for example, for the elimination of feed fluids in the event of quality deviations.

[0076] Illustratively, when the process is stopped, the fluid present in the system may be discharged through an outlet valve different from the outlet valve dedicated to the target component to avoid contamination / dilution of the target component already recovered. Alternatively, all outlet valves may be closed when the process is stopped, e.g., when the pump is fully decelerated.

[0077] Similar steps can be taken if air gets in (e.g., despite the air sensor) or if the air sensor fails. The process can be stopped, the system can be vented, and / or the cause (e.g., empty buffer reservoir, loose hose connection) can be eliminated. Thus, excessive air input into the stationary phase can be avoided.

[0078] The UV detector also allows the system to immediately detect if the flow path upstream of the membrane adsorber(s) does not contain UV-active material, so that rinse and wash volumes can be optimized.

[0079] In addition, some process parameters may be directly controlled by the upstream UV signal. For example, if the composition of the feed solution changes, the residence time may be shortened or washing and elution steps may be adjusted depending on the concentration of the feed. In another example, the loading rate may be adjusted as a function of the UV signal in the event of product-related fluctuations in the membrane adsorber (due to prior input of the binding capacity and / or titer of the target component in the feed), e.g., during the first cycle or perfusion.

[0080] For example, if the concentration of a continuously operated perfusion bioreactor changes, this can be recorded by a UV sensor before the stationary phase and processed (e.g., by algorithms, multiple linear regression, neural networks, or artificial intelligence) to adjust process parameters to the changed conditions, ensuring the safety of the chromatographic process and / or providing early warning of possible deviations.

[0081] If there is a UV sensor after the membrane adsorber(s), a comparison of the UV signals from the pre-membrane adsorber and the post-membrane adsorber can provide an assessment of the quality of the separation process. Furthermore, adaptive automated process control, such as variable regeneration steps or CIP steps, can be enabled based on the yield reduction seen by comparing the peak areas before and after the membrane adsorber.

[0082] The pre-membrane UV sensor also provides the capability of filter monitoring, if a filter is present. For example, if the UV signal changes significantly within a given time span, this may indicate a change in the feed composition or a ruptured / failed filter. In this case, downstream fluids may be pumped to waste, the process may be interrupted, and the filter may be replaced.

[0083] Furthermore, if a filter is installed in the inlet line dedicated to the feed fluid, two-stage chromatography may be possible in the system: the first stage is obtained by the filter and UV sensor before the membrane adsorber(s), while the second stage is obtained by the membrane adsorber(s) and UV sensor afterwards.

[0084] Finally, a pre-membrane UV sensor can provide a control function for comparable systems.

[0085] Overall, this system configuration with sensor technology in front of the stationary phase allows for integrated process control along the entire value chain.

[0086] The characteristics of the chromatography system, particularly the flow path, described above improve the chromatography process in terms of efficiency and quality. Illustratively, the membrane chromatography system described thus far is particularly suited to performing fast cycling chromatography, e.g., with cycles having durations of between about 3 and about 8 minutes with residence times of between about 10 and about 60 seconds, in view of the highly dynamic control of fluid flow within the flow path and the minimal dead volume. [Brief explanation of the drawings]

[0087] Detailed descriptions of exemplary embodiments are provided below with reference to the exemplary drawings. Other features will become apparent from the description, drawings, and claims. However, even if the embodiments are described separately, it should be understood that single features of different embodiments may be combined into further embodiments. [Figure 1] FIG. 1 shows a schematic diagram of an exemplary chromatography system. [Figure 2] Figure 2 shows a conceptual diagram of the inlet line in the flow channel. [Figure 3] FIG. 3 shows a plot of the maximum volume of the membrane adsorber without product loss as a function of several characteristics of the flow path. [Figure 4] FIG. 4 shows the optimization routine for determining the optimal switching times. [Figure 5] FIG. 5 shows plots of UV detection signal versus volume for different sampling rates of a UV filter in a chromatography system. [Figure 6] FIG. 6 shows a magnified plot of UV detection signal versus volume for different sampling rates of the UV filter in a chromatography system. [Figure 7] FIG. 7 shows plots of UV detection signal versus time for different configurations of filters in a chromatography system. [Figure 8] FIG. 8 shows plots of conductivity signal versus volume for different configurations of filters in a chromatography system. [Figure 9] FIG. 9 shows plots of normalized area versus normalized volume for different configurations of filters in a chromatographic system. [Figure 10] FIG. 10 shows plots of UV detection signal and conductivity signal versus time for different configurations of filters in a chromatography system. [Figure 11] FIG. 11 shows a schematic diagram of an exemplary chromatography system. [Figure 12] FIG. 12 shows another schematic diagram of an exemplary chromatography system. DETAILED DESCRIPTION OF THE INVENTION

[0088] In the following, the embodiments will be described in detail with reference to the drawings. It should be understood that various modifications can be made to the embodiments. Unless otherwise specified, one or more elements of one embodiment can be combined and used in another embodiment to form a new embodiment.

[0089] 1 shows a conceptual diagram of an exemplary chromatography system 100. Chromatography system 100 includes a flow path 110 and at least one membrane adsorber 200 as a stationary phase. Flow path 110 includes a conduit, such as a pipe and / or tubing, through which a fluid can flow, which includes a plurality of fluid-control components configured to regulate the flow of the fluid. Chromatography system 100 includes a control system (not shown) configured to manage at least some of the fluid-control components. Chromatography system 100 may be particularly suitable for membrane chromatography in bind-and-elute mode.

[0090] The dashed elements in Figures 1 and 2 are optional. The flow path 110 may initially include one or more inlet lines, which merge into a main line in the case of multiple inlet lines. An exemplary inlet line includes at least one inlet valve 120 configured to direct an input fluid, such as a supply fluid, buffer, or cleaning solution, into the flow path, as shown in detail in Figure 2. If the flow path 110 includes only one inlet line, the inlet line includes at least two inlet valves 120, one dedicated to the supply fluid and one for other input fluids. Each inlet valve 120 is configured to be connected to at least one input fluid source.

[0091] The inlet line may further include a pump 125, and an air sensor 130 configured to detect air in the inlet tubing may be installed between the inlet valve(s) 120 and the pump 125. After the pump 125, the inlet line may include a check valve 140, for example, if the switching time of the inlet valve(s) 120 is about 3 seconds or greater than about 3 seconds. After the check valve 140, other sensors 130, such as a pressure sensor and a flow meter, may be installed to monitor the inlet line.

[0092] If the inlet line is a line dedicated to the feed fluid, the feed inlet line may include a filter 150 configured to filter the feed fluid, for example, to exclude some particles, after the sensor 130 following the pump 125. In particular, the filter can be inserted directly into the feed inlet inline, i.e., without a valve. Alternatively, the filter 150 may be located in the main line. An analysis of the impact of filter location is provided below with reference to Figures 7-10.

[0093] If the flow path comprises multiple inlet lines, each inlet line may be provided at its end, i.e., before joining the main line with the other inlet lines, with a mixing valve 145. The provision of a mixing valve separates the different fluids from one another and prevents back-mixing, as also described with reference to Figure 10 below.

[0094] Returning to FIG. 1 , the flow path 110 comprises an inlet line(s) followed by a main line to which the membrane adsorber(s) 200 are connected. In one example, a single membrane adsorber 200 or a stack of membrane adsorber 200 on top of each other can be connected to the flow path 110. In another example, two membrane adsorber 200 or a stack of two membrane adsorber 200 may be connected in parallel to the flow path 110. The use of two membrane adsorber 200 or two stacks in parallel may enable a combination of capture and flow-through modes or an enhancement of the membrane volume, and therefore the binding capacity. The term "membrane volume" refers to the volume of a single membrane adsorber 200 or a stack of membrane adsorber 200 taking into account porosity. For example, 1 L of membrane volume may be the result of a 200 mL membrane layer and 800 mL of porosity.

[0095] The membrane adsorber(s) 200 may be connected to the flow path 110 by a (membrane) valve. The main line may branch into a path without a membrane adsorber 200 and one or two paths to which the membrane adsorber(s) 200 can be connected.

[0096] If the filter 150 is not located in the feed inlet line, the main line of the flow path 110 may be equipped with a filter 150 upstream of the membrane adsorber(s) 200. The filter 150 may be connected to the flow path by a (filter) valve. Thus, the main line may have two alternative branches, one equipped with the filter 150 and the other without the filter 150.

[0097] The main line of the flow path 110 may include one or more sensors 160 before the membrane adsorber(s) 200 (and after the filter 150, if present). In particular, a UV sensor may be placed before the membrane adsorber(s) 200 to provide monitoring capabilities and enable adaptive control of the system. Other sensors 160 may include pressure sensors, conductivity sensors, and pH sensors.

[0098] The main line of the flow path 110 comprises one or more sensors 170 after the membrane adsorber(s) 200. In particular, at least a UV sensor 170 is disposed between the membrane adsorber(s) 200 and an outlet valve, where the UV sensor is configured to detect whether the fluid coming from the membrane adsorber(s) 200 contains the target component and should therefore be directed to a product collection vessel or other outlet, e.g., waste. Other sensors 170 may include a pressure sensor, a conductivity sensor, and a pH sensor.

[0099] The signal generated by the UV sensor 170 is used by a control system to control an outlet valve located at the end of the flow path after the UV sensor 170. The sampling rate of the UV sensor 170 may be less than about 0.7 seconds, preferably less than about 0.5 seconds, and more preferably about 0.3 seconds or less. A description of the sampling rate is provided below with reference to Figures 5 and 6.

[0100] Flow path 110 includes at least two outlet valves, 180 and 185, and optionally additional outlet valves, each configured to be connected to a collection container. Outlet valve 180 may be connected to a target component collection container (and thus referred to as the "target component outlet valve"), while outlet valve 185 may be connected to a waste collection container (and thus referred to as the "waste outlet valve").

[0101] Flow path 110 may include a check valve 195 after waste outlet valve 185 if the switching time of waste outlet valve 185 is about 3 seconds or greater than about 3 seconds. In some examples, flow path 110 may include a check valve after each outlet valve if the switching time of the corresponding outlet valve is about 3 seconds or greater than about 3 seconds.

[0102] With respect to the design of a chromatography system, the portion of the flow path between the (post-membrane) UV sensor 170 and the output valve is of particular interest. The maximum membrane volume V that can be operated without losses is MAis coupled to the volumetric flow rate, and for fast non-kinetically limited stationary phases, the chromatographic system can be designed accordingly. This relationship can be derived from Equations (1)-(4) below:

number

number

number

number

[0103] During the ceremony, TIFF0007753360000006.tif8170 is the maximum volumetric flow rate without losses, d is the diameter, L is the length of the pipe between the UV sensor 170 and the output valve, and t tot is the total signal transit time. The total signal transit time consists of all time delays in signal transit from the moment the UV sensor detects the passage of the target component until the signal is acted upon, i.e., the target component outlet valve 180 opens. Equation (5) expresses the time t in terms of the time required by the control system for signal transit, sensor sampling rate, and valve switching time. tot 1 shows an exemplary decomposition of

number

[0104] Maximum volume flow rate V * is the unit time V * =MV·V MA It can be expressed as the number of membrane volumes per unit volume.

[0105] FIG. 3 shows the relationship between the pipe diameter d, the pipe length L, and the target component outlet valve switching time t at a volumetric flow rate of 5 membrane volumes per minute (MV=5 / min). switch The maximum membrane volume without loss of product as a function of volume is shown.

[0106] Increasing the valve switching time and shortening the pipe length increases the maximum possible volume V MA With larger pipe diameters, the maximum volume of stationary phase can be achieved with shorter tubing lengths due to the reduced flow rate.

[0107] t at MV=5 / min and pipe length 0.25m cont Assuming a time of 0.25 seconds for MA A tot As the total signal transit time increases, the available stationary phase volume and therefore the operating range of the system without product loss decreases. [Table 2]

[0108] From all the above considerations, it is clear that the variable t tot It can be seen that there is an interaction between L, d, and MV. Therefore, the following optimization routine can be applied:

number

[0109] Figure 4 shows the length / diameter and t of the pipe. tot , and therefore presents an optimization routine to determine the optimal switching time for the applicable stationary phase volume.

[0110] As noted above, the sampling rate, in combination with other parameters, serves to optimize the system to avoid product loss. Figures 5 and 6 show the effect of sampling rate alone on the performance of a chromatographic system.

[0111] FIG. 5 shows plots of UV detection signal versus volume for different sampling rates of a UV filter in a chromatography system.

[0112] As mentioned above, the presence of product / target molecules downstream of the membrane adsorber is typically detected by checking whether a given condition (the "valve switching condition") is met, i.e., whether adsorption at a defined wavelength (e.g., 280 nm) exceeds a given threshold, e.g., 0.05 AU. As long as the adsorption, detected by, e.g., a UV sensor, remains above the threshold, the product is collected by keeping the target component outlet valve 180 open.

[0113] Figure 5 shows the elution peak of BSA obtained with Sartobind® Q and indicates the cut points on the elution curve for two different scan rate values, 1 second and 0.3 seconds, where the UV sensor "recognizes" that adsorption has exceeded a threshold.

[0114] For different scan rate values, the cut points occur at different times / volumes, a difference clearly shown in Enlarged Figure 6. This condition is already met at approximately 2525 mL, but the UV sensor detects it with a delay in both cases. However, for a 45 L / h flow rate, detection occurs after less than 4 mL at the faster sampling rate, while detection occurs after more than 12 mL at the 1.0 s sampling rate. Higher flow rates result in a higher volumetric distance between cut points.

[0115] The hatched areas indicate the volume intervals in which the product is collected. Product loss can be calculated by taking the difference between the integral of the entire elution peak and the integral of each of its hatched fractions. [Table 3]

[0116] Therefore, a reduction in scan rate results in a reduction in product loss.

[0117] In addition to sampling rate, another parameter that has been considered in combination with other parameters with reference to Figures 3 and 4 is the switching time of the target component outlet valve 180. The effect of valve switching time alone on the performance of a chromatographic system is shown below with reference to five tests.

[0118] Because acetone absorbs light at 280 nm wavelength similarly to proteins and is therefore a suitable model, product elution is simulated in Tests 1, 2, and 3 using water and water / acetone (2-5% v / v). In Tests 4 and 5, analysis is performed using a Sartobind® Q loaded with 1 L of bovine serum albumin (BSA) (c = 3 g / L) eluted with 0.5 M NaCl. The valve switching condition is 0.1 AU for all experiments. Each test was performed at least three times, and appropriate fractions were removed and analyzed. The results are shown in the table below. [Table 4]

[0119] In Test 1 and Test 2, the switching time of outlet valve 180 and outlet valve 185 was set to about 3 seconds. In Test 2, check valve 195 was placed after waste outlet valve 185. In Test 3, Test 4, and Test 5, the switching time of outlet valve 180 and outlet valve 185 was set to about 0.5 seconds.

[0120] Compared to Test 2 and Test 3, Test 1 shows significantly lower mean signal intensity for the collected fractions. Additionally, the deviation between the extracted fractions was the highest of all tests at 33.4%. Implementation of check valve 185 resulted in a significantly higher concentration of 0.890 AU and a relative deviation of 3.1%, which is significantly lower relative to Test 1. Comparable performance to Test 2 is seen in Tests 3, 4, and 5.

[0121] Therefore, the valve switching time has a significant impact on reproducibility and product concentration, where the shorter the switching time, the better. However, due to safety aspects such as pressure generation in the system when pumping liquid at high volumetric flow rates, the valve switching time should not be too low, i.e., too close to 0 seconds. Therefore, to reduce backmixing and product loss, a valve switching time of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds can be selected.

[0122] Thus, outlet valve 180 and outlet valve 185 of system 100, and any optional additional outlet valves, are configured to have switching times of less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds. Alternatively, outlet valve 180 and outlet valve 185 may have switching times of about 3 seconds or greater, with at least waste outlet valve 185 having a check valve 195 disposed thereafter. If additional outlet valves are present, they may also have check valves disposed thereafter. Optionally, target component outlet valve 180 may also have a corresponding check valve 190.

[0123] Additionally, input valves 120 may be controlled to have switching times less than about 3 seconds, preferably less than about 1 second, and most preferably equal to about 0.5 seconds. Alternatively, input valves 120 may have switching times greater than about 3 seconds, and each inlet line may be equipped with a check valve 140. The same concept applies to all valves in flow path 110 of system 100.

[0124] Another aspect that may be considered in the design of the system is the location, if any, of the filter 150. Depending on the location of the pre-filter, there are differences in backmixing, which are discussed below.

[0125] Figure 7 shows plots of UV detection signal versus time for different configurations of filters in a chromatography system. The three configurations are as follows: A) Filter connected to the main line by a filter valve with a switching time of approximately 3 seconds, with only the feed fluid flowing through (dotted line). B) Filter inserted directly into the main line, all fluid flow-through (solid line) C) Filter inserted directly into the supply fluid inlet line (dashed line).

[0126] The same test procedure was performed for each configuration, consisting of equilibration with water, loading with a 2-5% (v / v) water / acetone mixture, washing with water, accidental elution, and regeneration with water. Elution was performed by gradually increasing the water / acetone mixture until a signal of 0.2 AU was reached, and then regeneration was initiated.

[0127] Configuration A shows a signal drop at approximately 0.8 minutes. Further along in the test, an elution peak is identified at 3 minutes, and a further peak at 3.7 minutes. The concentration drop during the loading process can be explained by backmixing in the filter between the feed solution and water due to the volume of the piping between the pump and the filter and the switching time of the filter valve. This mixing results in an undesirable dynamic concentration profile during the loading step, which adversely affects the binding properties of the stationary phase.

[0128] In the case of configuration B, the signal does not decrease during loading, but does not reach a constant value. The elution peak is clearly broader than that of configuration A, and a second peak is again visible. As all fluids pass through the filter, each fluid mixes with residues from previously filtered fluids. This mixing results in concentration changes that adversely affect the performance of the system.

[0129] In the case of configuration C, the signal shows a sharp rise in the loading phase and a sharp peak in the elution phase. Again, a second peak is identified. All things considered, this configuration shows the best results for hydrodynamics / backmixing: the signal is always stable and shows a narrow elution peak.

[0130] Configuration C has the lowest backmixing volume, while configuration A has a narrow peak but also has the option of in-line dilution in the system. The performance of configuration A can be improved by reducing the switching time of the filter valve, as shown in Figure 8.

[0131] Figure 8 shows plots of conductivity signal versus volume for different configurations of filters in a chromatography system. In particular, the solid line represents configuration C, in which the filter is installed directly in the feed inlet line (also referred to as an "in-line filter"). The dashed line represents configuration A, in which the filter is connected to the main line by two filter valves (also referred to as an "on-line filter"), with the modification that the filter valve switching time is approximately 0.5 seconds. Finally, the dotted line also represents a configuration with an on-line filter and a switching time of approximately 0.5 seconds.

[0132] All curves were obtained by pumping different water and water / acetone mixtures at 45 L / hr to each filter location. It can be seen that the performance of the online filter with a 0.5 second valve switching time is comparable to that of the in-line filter. This can also be seen in Figure 9, which shows a plot of the conductivity-normalized area versus the signal area centroid-normalized volume for the in-line and online filters.

[0133] The poor performance of configuration B relative to configuration C can be theoretically explained using the equilibrium variance model expressed in Equation 7 below, where c i is the concentration of the component in the feed fluid, and u int is the linear velocity of the feed fluid, and D ax is the axial dispersion coefficient, which is the sum of the contributions of axial molecular diffusion and eddy diffusion.

[0134] In a typical velocity or equilibrium dispersion model, concentration varies with time, here calculated by the change in concentration across the length (i.e., the dimension in the direction of flow). The change in concentration over time is further separated into convective and diffusive / dispersive mass transfer. The convective term describes the concentrated flow into the next length interval by the linear velocity and the change in length. The diffusive / dispersive mass transfer is described by the axial dispersion coefficient. The size / effect of axial dispersion is described by the value of the axial dispersion coefficient and the change in concentration through the cross-sectional area, which is represented by the second derivative of the concentration with respect to length, which is comparable to the second Fick's law. The second derivative of the concentration with respect to length describes backmixing.

[0135] In other words, the change in concentration over time is due to convective transport via the linear velocity and backmixing via the axial dispersion coefficient. More precisely, the longer a volume is swept by the fluid, the greater the effect of backmixing. The locally considered concentration change ∂ 2 c i increases with increasing total temporal concentration change over the length times the axial dispersion coefficient, as shown by Equation 8.

number

number

[0136] When the filter is permanently and exclusively flushed by the feed, as in configuration C, the concentration change ∂c i Since / ∂t is zero in the filter, backmixing in the system is significantly reduced.

[0137] If the filter is not installed directly in an inlet line used exclusively for the feed, it should be flushed only by the feed. Furthermore, the filter should be permanently filled with the feed, i.e., during the entire cycle. Therefore, the switching time of the filter valve must be adjusted according to the volume of the feed returning to the filter valve to avoid concentration gradients.

[0138] As shown in Figure 7, a second elution peak is detected in all configurations A, B, and C. Figure 10 shows the conductivity signal (dashed line) in addition to the UV signal (solid line) for configuration A. The conductivity signal features a small peak at the beginning of loading, i.e., a salt signal that reduces the binding capacity of the stationary phase. This undesirable behavior occurs due to the merging of multiple inlet lines without effectively preventing backmixing.

[0139] Providing a mixing valve 145 in each inlet line creates a dedicated mixing point, separating the different media from each other, so that the concentration gradient is zero up to the mixing point, which means that backmixing is not possible (see Equation 7). In the case of an online filter, the presence of a mixing valve also allows for a reduction in the distance between the point where the inlet lines meet and the location of the filter.

[0140] A further means for reducing backmixing relates to the general design of the flow path from inlet to outlet: a flow path with minimal bends and turns, or in other words, a flow path that is as straight as possible, reduces the dead volume of the flow path and therefore reduces backmixing.

[0141] 11 and 12 show two examples of chromatography systems in which one or more of the previously illustrated means for optimizing a chromatography process are implemented.

[0142] 11 shows an exemplary implementation of a chromatography system 900. System 900 has three inlet lines, each containing multiple inlet valves 901 / 903 / 905, air sensors 907 / 909 / 911, pumps 913 / 915 / 917, check valves 919 / 921 / 923, pressure sensors 925 / 927 / 929, and flow meters 931 / 933 / 935. One inlet line is for feed, another inlet line is for water to perform in-line dilution, and the final inlet line is for buffer.

[0143] A filter 940 is provided along the flow path after the three inlet lines converge. In particular, the flow path is provided with two filter valves 942, 944 for connecting the filter 940. The flow path further includes a filter bypass valve 946 and a drain valve 948 that provide alternate paths for the incoming fluid.

[0144] Subsequently, two membrane adsorber (or two stacks) 960, 961 are each connected to the flow path by two membrane valves 962, 963 and 964, 965, respectively. The volume of each membrane adsorber / each stack is 150 mL. Sensor settings 950 and 970 for measuring pressure, conductivity, pH and adsorption are implemented before and after the membrane adsorber 960, 961. The flow path further comprises a membrane bypass valve 966.

[0145] Finally, four outlet valves 980, 985, 990, and 995 are provided to discharge waste, final product, and various process intermediates.

[0146] All valves in the system 900 are controlled to have switching times of less than about 3 seconds, except for a number of inlet valves 901, 903, 905, which are then implemented with check valves 919, 921, 923, respectively.

[0147] As noted above, due to the smaller stationary phase volume and lower binding capacity per cycle compared to conventional column chromatography, a greater number of membrane chromatography cycles are required to process a given volume of feed, however, each cycle is shorter, as can be seen from the table below for Protein A chromatography performed using system 900. [Table 5]

[0148] The above values ​​refer to a cycle of membrane chromatography in bind-and-elute mode with a fixed transition from one step to the next. The volume of each step is expressed in units of membrane volume, which is 150 mL. The residence time of the protein-containing solution on the membrane is less than 20 seconds, as convection is primarily responsible for mass transport, resulting in more efficient adsorption compared to the diffusion mechanism that predominates on the resin. The dynamic binding time is approximately 20 g / L.

[0149] A membrane chromatography cycle performed in system 900 of FIG. 9 lasts approximately 7 minutes, while a corresponding cycle of column chromatography, as previously mentioned, lasts over 4 hours.

[0150] 12 shows another exemplary implementation of a chromatography system 1000. The chromatography system 1000 includes two inlet lines, each containing multiple inlet valves 1001 / 1003, air sensors 1005 / 1007, pumps 1009 / 1011, pressure sensors 1013 / 1015, flow meters 1017 / 1019, and mixing valves 1021 / 1023. One inlet line is for the feed and the other is for the buffer.

[0151] A filter 1040 is provided along the flow path after the two inlet lines join. In particular, the flow path is provided with two filter valves 1042, 1044 for connecting the filter 1040. The flow path further includes a filter bypass valve 1046 and a drain valve 1048 that provide alternate paths for the incoming fluid.

[0152] Subsequently, two membrane adsorber (or two stacks) 1060, 1061 are each connected to the flow path by two membrane valves 1062, 1063 and 1064, 1065, respectively. The volume of each membrane adsorber / each stack is 150 mL. Sensor settings 1050 and 1070 for measuring pressure, conductivity, pH and adsorption are implemented before and after the membrane adsorber 1060, 1061. The flow path further comprises a membrane bypass valve 1066.

[0153] Finally, four outlet valves 1080, 1085, 1090, and 1095 are provided to discharge waste, final product, and various process intermediates.

[0154] All valves in the system 1000 are controlled to have switching times of less than about 3 seconds.

[0155] The duration of each phase of a Protein A chromatography cycle performed using System 1000 is reported in the following table: [Table 6]

[0156] The above values ​​refer to cycles of membrane chromatography in bind and elute mode with conditional transition from one step to the next, for example, transition to the next step when a given UV adsorption or conductivity value is reached.

[0157] A membrane chromatography cycle performed in the system 1000 of Figure 10 lasts approximately 4 minutes, and thus the system 1000 is faster than the system 900 shown in Figure 9. This is in part because, as previously explained, the presence of the mixing valve allows for a shorter flow path between the inlet line and the filter. Additionally, conditional transfers shorten the cycle.

[0158] The reduction in backmixing achieved by the structural design and / or flow control means discussed above results in lower peak broadening of the eluted fractions and therefore higher product concentrations in the eluate. Thus, there is no buildup of dilution over many cycles and the chromatographic process is improved in terms of efficiency and quality.

Claims

1. A chromatography system configured to process a feed fluid containing a plurality of components, wherein at least one component of the plurality of components of the feed fluid is a target component, and wherein the chromatography system: a flow path including a plurality of fluid control components configured to control fluid flow; a stationary phase, said stationary phase being at least one membrane adsorber connected to said flow path, said stationary phase configured to isolate said target component; Equipped with The plurality of fluid control components include: a first outlet valve connected to the at least one membrane adsorber and configured to be connected to a target component collection vessel; a second outlet valve connected to the at least one membrane adsorber and configured to be connected to a waste collection container; Equipped with A chromatography system, wherein the first outlet valve and the second outlet valve have a switching time of less than 3 seconds.

2. A chromatography system configured to process a feed fluid containing a plurality of components, wherein at least one component of the plurality of components of the feed fluid is a target component, and wherein the chromatography system: a flow path including a plurality of fluid control components configured to control fluid flow; a stationary phase, said stationary phase being at least one membrane adsorber connected to said flow path, said stationary phase configured to isolate said target component; Equipped with The plurality of fluid control components include: a first outlet valve connected to the at least one membrane adsorber and configured to be connected to a target component collection vessel; a second outlet valve connected to the at least one membrane adsorber and configured to be connected to a waste collection container; a check valve disposed after the second outlet valve; Equipped with A chromatography system, wherein the second outlet valve has a switching time of 3 seconds or greater.

3. The plurality of fluid control components include: a first inlet valve configured to be connected to a supply fluid source; a second inlet valve configured to be connected to a buffer source; Further provided with 3. The chromatography system of claim 1, wherein the first inlet valve and the second inlet valve have a switching time of less than 3 seconds.

4. The plurality of fluid control components include: a first inlet valve configured to be connected to a supply fluid source; a second inlet valve configured to be connected to a buffer source; at least one inlet check valve disposed after the first inlet valve and the second inlet valve; Further provided with 3. The chromatography system of claim 1, wherein the first inlet valve and the second inlet valve have a switching time of 3 seconds or more.

5. The chromatography system of any one of claims 1 to 4, wherein the plurality of fluid control components further comprises a filter configured to filter only the feed fluid.

6. 6. The chromatography system of claim 5, wherein the at least one membrane adsorber has a first pore size and the filter has a second pore size, the second pore size being smaller than the first pore size.

7. 7. The chromatography system of claim 1, wherein the plurality of fluid control components further comprises an adsorption detector disposed after the at least one membrane adsorber, and the sampling period of the adsorption detector is less than 0.7 seconds.

8. 8. The chromatography system of claim 1, wherein the plurality of fluid control components further comprises an adsorption detector positioned before the at least one membrane adsorber.

9. The chromatography system of claim 1 , wherein the first outlet valve and the second outlet valve have a switching time of less than 1 second.

10. 10. The chromatography system of claim 1, wherein the first outlet valve and the second outlet valve have a switching time equal to 0.5 seconds.

11. The chromatography system of claim 3 , wherein the first inlet valve and the second inlet valve have a switching time of less than 1 second.

12. 4. The chromatography system of claim 3, wherein the first inlet valve and the second inlet valve have a switching time equal to 0.5 seconds.

13. 8. The chromatography system of claim 7, wherein the adsorption detector has a sampling period of less than 0.5 seconds.

14. 8. The chromatography system of claim 7, wherein the adsorption detector has a sampling period of 0.3 seconds or less.

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