Preparative chromatography system and method for chromatography separation

Abstract

To provide a preparative chromatography system and an operation method which overcome the disadvantages of a preparative chromatography system of a prior art.SOLUTION: There are provided preparative chromatography systems 200, 500 and 800, and chromatographic processing 400 and 700 which are adapted to an iteration cycle of a chromatography capacity. The systems 200, 500 and 800 include at least two upstream pumps 203a, 803a, 203b, and 803b, and a separate flow channel 220 to the chromatography devices 200, 500 and 800 from a treatment liquid source. The systems 200, 500 and 800 are configured to provide a separate treatment liquid to the chromatography device, and to prime one flow channel 220 with one treatment liquid, while minimizing hold-up capacities of the systems 200, 500 and 800.SELECTED DRAWING: Figure 2a

Description

Technical Field

[0001] The present invention relates to a preparative chromatography system, a method of operating a preparative chromatography system, and a chromatography process adapted to an iterative cycle of chromatography volume. In particular, the present invention relates to systems and methods optimized for relatively small volume chromatography devices, typically membrane chromatography devices.

Background Art

[0002] Chromatography is today a major industrial-scale method for providing high-purity substances, particularly high-purity proteins, in the pharmaceutical field. Historically, separation and purification have been achieved primarily by conventional packed-bed chromatography utilizing columns of porous beads. Porous bead-based systems have well-known drawbacks with respect to the high flow rates desirable for obtaining high system throughput, drawbacks resulting from separation processes in which diffusion is limited. At least in part to address these problems, alternatives to conventional porous bead-based systems have been introduced and developed over the past decade. These alternative technologies include the use of monoliths and membranes. Flow through such systems is typically convective rather than diffusive, and thus is less affected by flow than porous bead-based systems and can be run at considerably higher flow rates. However, until recently, the binding capacity of membrane-based systems has been a limiting factor. The introduction of fiber-based as a chromatography medium has significantly reduced the drawbacks of previous membrane-based systems. Patent Document 1 discloses a polymer fiber-based chromatography medium. Such documents, and all other references indicated herein (whether shown with respect to written disclosures, products, etc.), are incorporated by reference to the maximum extent permitted.

[0003] All chromatographic systems exhibit a "dead volume", defined as the extra volume experienced by the solute as it passes through the chromatographic system, particularly the non-swept volume exposed to the mobile phase flow. The term "dead volume" is used in various meanings and is not always correct and is regarded as an outdated term. The related term "hold-up volume" has been established as a more clearly defined alternative and is equal to the retention volume of non-retained compounds and is, in most cases, the most relevant parameter to consider. The hold-up volume includes the volumes contributed by the sample injector, detector, pump, valves, tubes, connectors, etc. If the hold-up volume is large compared to the volume of the chromatographic device, the peak resolution decreases and the waste of buffer and feed increases.

[0004] The control of the hold-up volume is a concern in the design of chromatographic systems and has generally been regarded as something to be carefully addressed rather than an overly complex design choice. However, with the emergence of new chromatographic media such as fibers, the volume of the active component, now called the chromatographic device, has become much smaller than the volume of the column / cartridge in conventional porous bead-based systems. As a result, the volume of the chromatographic device is of the same order as, or in some cases even smaller than, the volume of the tubes, valves, pumps, etc. that transport the feed and buffer to the chromatographic device. In these new chromatographic systems, the hold-up volume has emerged as an important factor. Furthermore, in preparative chromatographic systems where the advantages of new separation techniques are most effectively utilized, the process is repeated many times and the adverse effects of the current relatively large hold-up volume become even more pronounced. For example, the previous design principles and general considerations when selecting tubes are not sufficient for new small chromatographic devices. Therefore, there is a need for high-flow preparative chromatographic systems with a small hold-up volume that can handle high flow rates.

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] Pamphlet of International Publication No. 2018 / 011600 [Summary of the Invention] [Problems to be Solved by the Invention]

[0006] An object of the present invention is to provide a preparative chromatography system and an operation method that overcome the drawbacks of prior art preparative chromatography systems. [Means for Solving the Problems]

[0007] This is achieved by a method of operating a preparative chromatography system defined in claim 1 and a preparative chromatography system defined in claim 13.

[0008] According to one aspect of the present invention, there is provided a method of performing preparative chromatography for operating a preparative chromatography system. The preparative chromatography system is configured to operate with at least a first processing liquid and a second processing liquid. The preparative chromatography system includes at least one chromatography device, a first processing liquid source, and a second processing liquid source. The first upper flow path from the first processing liquid source to the chromatography device, the second upper flow path from the second processing liquid source to the chromatography device, a first pump provided in the first upper flow path and configured to provide the first processing liquid to the chromatography device, and a second pump provided in the second upper flow path and configured to provide the second processing liquid to the chromatography device. The first upper flow path merges with the second upper flow path at a mixing point. The mixing point is downstream of the first pump and the second pump and upstream of the chromatography device. The method includes: - Operating the first pump to prime the first upper flow path with the first processing liquid, and operating the first pump to provide the first processing liquid to the chromatography device after the priming step; - Operating the second pump to prime the second upper flow path with the second processing liquid, and after the priming step, operating the second pump to supply the second processing liquid to the chromatography apparatus; and the like.

[0009] According to an embodiment of the present invention, the preparative chromatography system is configured to operate with at least three processing liquids including a supply liquid, a cleaning liquid, and an eluent. The preparative chromatography system includes a supply source, a cleaning source, and an eluent source, and provides an upper supply flow path from the supply source to the chromatography apparatus, an upper cleaning flow path from the cleaning source to the chromatography apparatus, and an upper eluent flow path from the eluent source to the chromatography apparatus. The method - Priming the upper supply flow path with the supply liquid provided from the supply source; - Priming the upper cleaning flow path with the cleaning liquid provided from the cleaning source; - Priming the upper eluent flow path with the eluent provided from the eluent source; and includes a previous stage including the above. The method - Balancing the chromatography apparatus by providing a cleaning liquid from the cleaning source to the chromatography apparatus through the upper cleaning flow path; - Loading the supply liquid into the chromatography apparatus by providing the supply liquid from the cleaning source to the chromatography apparatus through the upper supply flow path; - Cleaning the chromatography apparatus by providing a cleaning liquid from the cleaning source to the chromatography apparatus through the upper cleaning flow path; - Performing elution in the chromatography apparatus by providing an eluent liquid provided from the eluent supply source to the chromatography apparatus through the upper eluent flow path; - Re-balancing the chromatography apparatus by providing a cleaning liquid from the cleaning source to the chromatography apparatus through the upper cleaning flow path; includes an active fractionation step including a processing cycle involving

[0010] According to one embodiment of the present invention, a preparative chromatography system is configured to operate with at least three processing fluids, namely a feed solution, a cleaning solution, and an eluent, and the preparative chromatography system - a feed source connected to a feed pump, a cleaning source connected to a cleaning pump, and an eluent source connected to an eluent pump, and - a first chromatography device associated with a first monitoring unit, a second chromatography device associated with a second monitoring unit, and a third chromatography device associated with a third monitoring unit, and - a first upper feed flow path provided from the feed source to the first chromatography device, a second upper feed flow path provided from the feed source to the second chromatography device, and a third upper feed flow path provided from the feed source to the third chromatography device, wherein the upper feed flow paths are bifurcated downstream of the feed pump, the first, second, and third upper feed flow paths, and - a first upper cleaning flow path provided from the cleaning source to the first chromatography device, a second upper cleaning flow path provided from the cleaning source to the second chromatography device, and a third upper cleaning flow path provided from the cleaning source to the third chromatography device, wherein the upper cleaning flow paths are bifurcated downstream of the cleaning pump, the first, second, and third upper cleaning flow paths, and - a first upper eluent flow path provided from the eluent source to the first chromatography device, a second upper eluent flow path provided from the eluent source to the second chromatography device, and a third upper eluent flow path provided from the eluent source to the third chromatography device, wherein the upper eluent flow paths are bifurcated downstream of the eluent pump, the first, second, and third upper eluent flow paths, and comprises The method is executed at least partially simultaneously - By operating the cleaning pump, one of the first, second, or third chromatography apparatuses is equilibrated or cleaned with the cleaning liquid provided from the cleaning source through one of the first upper cleaning flow paths; - By operating the supply pump, another one of the first, second, or third chromatography apparatuses is loaded with the supply liquid provided from the supply source through one of the upper supply flow paths; - By operating the eluent pump to provide the eluent from the eluent source through one of the upper eluent flow paths, performing elution in the remaining part of the first, second, or third chromatography apparatus, wherein the eluate is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit; including an active fractionation step.

[0011] According to one embodiment of the present invention, the fractionation chromatography system is configured to operate with at least three processing liquids of the supply liquid, the cleaning liquid, and the eluent, and the fractionation chromatography system a supply source, a cleaning source, and an eluent source; a first pump, a second pump, a first waste container, and a common valve connected to the chromatography apparatus, wherein the fractionation chromatography system has a first chromatography flow path that passes the first pump from at least one of the processing liquid sources to the common valve, and a second chromatography flow path that passes the second pump from at least one of the processing liquid sources to the common valve, the first pump, the second pump, the first waste container, and the common valve; a first waste flow path that passes the first pump from at least one of the processing liquid sources to the first waste container, and a second waste liquid flow path that passes the second pump from at least one of the processing liquid sources to the common valve; comprising; In the steps of the method, While the first pump is engaged in providing a processing liquid to the chromatography apparatus, the second pump is at least partially simultaneously engaged in providing the processing liquid to the waste container via one of the second waste flow paths.

[0012] According to an embodiment of the present invention, the fractionation chromatography system includes an upper monitoring unit configured in front of the flow direction to the first waste container, and the upper monitoring unit is configured to analyze the contents in the first waste flow path and the second waste flow path. In the steps of the method, the duration of at least one of the priming operations is determined based on the output of the measuring unit.

[0013] According to an aspect of the present invention, a fractionation chromatography system is provided. The fractionation chromatography system is configured to operate with at least a first processing liquid and a second processing liquid. The fractionation chromatography system includes at least one chromatography apparatus, a first processing liquid source, and a second processing liquid source. A first upper flow path from the first processing liquid source to the chromatography apparatus, a second upper flow path from the second processing liquid source to the chromatography apparatus, a first pump provided in the first upper flow path and configured to provide the first processing liquid to the chromatography apparatus, and a second pump provided in the second upper flow path and configured to provide the second processing liquid to the chromatography apparatus are provided. The first upper flow path merges with the second upper flow path at a mixing point. The mixing point is downstream of the first pump and the second pump and upstream of the chromatography apparatus. The system In the step of priming the first upper flow path with the first processing liquid that operates the first pump, and in the step performed after the priming step, the first pump is operated to provide the first processing liquid to the chromatography apparatus. In the step of priming the second upper flow path with the second processing liquid, the second pump is operated, and in the step performed after the priming step, the second pump is operated to provide the second processing liquid to the chromatography apparatus. is configured as follows.

[0014] According to one embodiment of the present invention, the preparative chromatography system is configured to operate with at least three processing liquids, namely a feed liquid, a cleaning liquid, and an eluent, and the preparative chromatography system includes a feed source, a cleaning source, and an eluent source, which provide an upper feed flow path from the feed source to the chromatography device, an upper cleaning flow path from the cleaning source to the chromatography device, and an upper eluent flow path from the eluent source to the chromatography device. The preparative chromatography system - a step of priming the upper feed flow path with the feed liquid provided from the feed source; - a step of priming the upper cleaning flow path with the cleaning liquid provided from the cleaning source; - a step of priming the upper eluent flow path with the eluent provided from the eluent source; is configured to operate the previous steps; The preparative chromatography system - a step of equilibrating the chromatography device by providing a cleaning liquid from the cleaning source to the chromatography device via the upper cleaning flow path; - a step of loading the feed liquid into the chromatography device by providing the feed liquid from the cleaning source to the chromatography device via the upper feed flow path; - a step of cleaning the chromatography device by providing a cleaning liquid from the cleaning source to the chromatography device via the upper cleaning flow path; - a step of performing elution in the chromatography device by providing an eluent liquid provided from the eluent supply source to the chromatography device via the upper eluent flow path; - a step of re-equilibrating the chromatography device by providing a cleaning liquid from the cleaning source to the chromatography device via the upper cleaning flow path; is configured to operate an active preparative step including a processing cycle having the above steps.

[0015] According to one embodiment, a preparative chromatography system includes a supply pump provided in an upper supply channel and configured to provide a supply liquid to a chromatography device, and an upper supply valve provided downstream of the supply pump in the upper supply channel, a cleaning pump provided in an upper cleaning channel and configured to provide a cleaning liquid to the chromatography device, and an upper cleaning valve provided downstream of the cleaning pump in the cleaning channel, an eluent pump provided in an upper eluent channel and configured to provide an eluent to the chromatography device, and an upper eluent valve provided downstream of the eluent pump in the eluent channel, a monitoring unit provided downstream of the chromatography device, and a lower valve provided downstream of the monitoring unit and configured to connect to either a waste container or a fraction collector. The preparative chromatography system is provided with: The preparative chromatography system includes - a step of priming the upper supply channel by controlling the supply pump and the upper supply valve to load a supply liquid into the supply channel, during which the upper cleaning valve and the upper eluent valve are closed; - a step of priming the upper cleaning channel by controlling the cleaning pump and the upper cleaning valve to load a cleaning liquid into the cleaning channel, during which the upper supply valve and the upper eluent valve are closed; - a step of priming the upper eluent channel by controlling the eluent pump and the upper eluent valve to load an eluent into the eluent channel, during which the upper cleaning valve and the upper supply valve are closed; and is configured to operate in the previous steps. The preparative chromatography system includes - Controlling a cleaning pump and an upper cleaning valve to load a cleaning liquid into and through a chromatography device to equilibrate the chromatography device, wherein the lower valve is set to a waste container and the upper supply valve and the upper eluent valve are closed during this step; - Controlling a supply pump and an upper supply valve to load a supply liquid into and through a chromatography device to load the supply liquid into the chromatography device, wherein the lower valve is set to a waste container and the upper cleaning valve and the upper eluent valve are closed during this step; - Controlling a cleaning pump and an upper cleaning valve to load a cleaning liquid into and through a chromatography device to clean the chromatography device, wherein the lower valve is set to a waste container and the upper supply valve and the upper eluent valve are closed during this step; - Controlling an eluent pump and an upper eluent valve to load an eluent liquid into and through a chromatography device to perform elution in the chromatography device, wherein the lower valve is set to either a waste container or a fraction collector, the setting depending on the content of the eluate detected by a monitoring unit, and the upper cleaning valve and the upper supply valve are closed during this step; - Controlling a cleaning pump and an upper cleaning valve to load a cleaning liquid into and through a chromatography device to re-equilibrate the chromatography device, wherein the lower valve is set to a waste container and the upper supply valve and the upper eluent valve are closed during this step; configured to operate an active fractionation stage including a processing cycle having the above.

[0016] According to one embodiment, the fractionation chromatography system is configured to operate with at least three processing liquids, namely a supply liquid, a cleaning liquid, and an eluent liquid. The fractionation chromatography system - A supply source connected to a supply pump, a cleaning source connected to a cleaning pump, and an eluent source connected to an eluent pump, - A first chromatography device related to a first monitoring unit, a second chromatography device related to a second monitoring unit, and a third chromatography device related to a third monitoring unit, - A first upper supply flow path provided from the supply source to the first chromatography device, a second upper supply flow path provided from the supply source to the second chromatography device, and a third upper supply flow path provided from the supply source to the third chromatography device, wherein the upper supply flow paths branch downstream of the supply pump, the first, second, and third upper supply flow paths, - A first upper cleaning flow path provided from the cleaning source to the first chromatography device, a second upper cleaning flow path provided from the cleaning source to the second chromatography device, and a third upper cleaning flow path provided from the cleaning source to the third chromatography device, wherein the upper cleaning flow paths branch downstream of the cleaning pump, the first, second, and third upper cleaning flow paths, - A first upper eluent flow path provided from the eluent source to the first chromatography device, a second upper eluent flow path provided from the eluent source to the second chromatography device, and a third upper eluent flow path provided from the eluent source to the third chromatography device, wherein the upper eluent flow paths branch downstream of the eluent pump, the first, second, and third upper eluent flow paths, comprising, The preparative chromatography system is, - By operating the cleaning pump, equilibrating or cleaning one of the first, second, or third chromatography devices with a cleaning liquid provided from the cleaning source via one of the first upper cleaning flow paths, - By operating the supply pump, loading a supply liquid provided from the supply source via one of the upper supply flow paths into another one of the first, second, or third chromatography devices, - By operating the eluent pump to provide eluent from an eluent source through one of the upper eluent flow paths, performing elution in the remaining part of the first, second, or third chromatography device, and directing the eluate to a waste container or fraction collector according to the content of the eluate detected by the monitoring unit. It is configured to operate an active fractionation step including a processing cycle having

[0017] According to one embodiment, the fractionation chromatography system - The supply pump is connected to the first four-way valve, the washing pump is connected to the second four-way valve, and the eluent pump is connected to the third four-way valve. - The first four-way valve is connected to the fourth, fifth, and sixth four-way valves. - The second four-way valve is connected to the fourth, fifth, and sixth four-way valves. - The third four-way valve is connected to the fourth, fifth, and sixth four-way valves, and - The fourth four-way valve is connected to the first chromatography device, the fifth four-way valve is connected to the second chromatography device, and the sixth four-way valve is connected to the third chromatography device. It is configured according to The fractionation chromatography system The supply liquid is from the supply source - By setting the first four-way valve to direct the flow to the fourth four-way valve and setting the fourth four-way valve to direct the flow to the first chromatography device, to the first chromatography device through the first upper supply flow path. - By setting the first four-way valve to direct the flow to the fifth four-way valve and setting the fifth four-way valve to direct the flow to the second chromatography device, to the second chromatography device through the second upper supply flow path, and - By setting the first four-way valve to direct the flow towards the sixth four-way valve and setting the sixth four-way valve to direct the flow towards the third chromatography device, the third chromatography device is supplied via the third upper supply channel, provided, with the cleaning liquid from the cleaning source - By setting the second four-way valve to direct the flow towards the fourth four-way valve and setting the fourth four-way valve to direct the flow towards the first chromatography device, the first chromatography device is supplied via the first upper cleaning channel, - By setting the second four-way valve to direct the flow towards the fifth four-way valve and setting the fifth four-way valve to direct the flow towards the second chromatography device, the second chromatography device is supplied via the second upper cleaning channel, and, - By setting the second four-way valve to direct the flow towards the sixth four-way valve and setting the sixth four-way valve to direct the flow towards the third chromatography device, the third chromatography device is supplied via the third upper cleaning channel, provided, with the eluent from the eluent source - By setting the third four-way valve to direct the flow towards the fourth four-way valve and setting the fourth four-way valve to direct the flow towards the first chromatography device, the first chromatography device is supplied via the first upper eluent channel, - By setting the third four-way valve to direct the flow towards the fifth four-way valve and setting the fifth four-way valve to direct the flow towards the second chromatography device, the second chromatography device is supplied via the second upper eluent channel, and, - By setting the third four-way valve to direct the flow towards the sixth four-way valve and setting the sixth four-way valve to direct the flow towards the third chromatography device, the third chromatography device is supplied via the third upper eluent channel, provided, and is configured as such.

[0018] According to one embodiment, the preparative chromatography system is configured to operate with at least three processing liquids, namely a supply liquid, a cleaning liquid, and an eluent, and the preparative chromatography system - A supply source, a cleaning source, and an eluent source, and - A first pump, a second pump, a first waste container, and a common valve connected to a chromatography device, and the preparative chromatography system includes - A first chromatography flow path through which the first pump passes from at least one of the processing liquid sources to the common valve, and a second chromatography flow path through which the second pump passes from at least one of the processing liquid sources to the common valve, and - A first waste flow path through which the first pump passes from at least one of the processing liquid sources to the first waste container, and a second waste liquid flow path through which the second pump passes from at least one of the processing liquid sources to the common valve. During the processing cycle, the preparative chromatography system is configured while the first pump is engaged in providing the processing liquid to the chromatography device, and the second pump is at least partially simultaneously engaged in providing the processing liquid to the waste container through one of the second waste flow paths.

[0019] According to one embodiment, the preparative chromatography system is configured to execute a predetermined chromatography device sequence that defines the order of the processing liquid passing through the chromatography device. In one step, one of the first pump and the second pump provides the processing liquid to the chromatography device, and at least partially simultaneously, the other of the first pump and the second pump primes the associated flow path with the processing liquid that orders the next processing liquid to be provided to the chromatography device according to the chromatography device.

[0020] According to one embodiment, the preparative chromatography system - Operating a first pump to provide a cleaning liquid from a cleaning source to a chromatography apparatus through a first cleaning chromatography flow path, thereby equilibrating the chromatography apparatus, and operating a second pump to provide a supply liquid from a supply source to a first waste container through a second supply waste flow path, thereby at least partially simultaneously priming the second pump with the supply liquid; - Operating a second pump to provide a supply liquid from a supply source to a chromatography apparatus through a second supply chromatography flow path, thereby loading a supply onto the chromatography apparatus, and operating a first pump to provide a cleaning liquid from a cleaning source to a first waste container through a first cleaning waste flow path, thereby at least partially simultaneously priming at least the first pump with the cleaning liquid; - Operating a first pump to provide a cleaning liquid from a cleaning source to a chromatography apparatus through a first cleaning chromatography flow path, thereby cleaning the chromatography apparatus, and operating a second pump to provide a cleaning liquid from a cleaning source to a first waste liquid container through a second cleaning waste liquid flow path, thereby at least partially simultaneously priming at least the second pump with the cleaning liquid; - Operating a second pump to provide a cleaning liquid from a cleaning source to a chromatography apparatus through a second cleaning chromatography flow path, thereby continuing to clean the chromatography apparatus, and operating a first pump to provide an eluent liquid from an eluent source to a first waste container through a first eluent waste flow path, thereby at least partially simultaneously priming at least the first pump with the eluent liquid; - Operating the first pump to provide an eluent to the chromatography apparatus from an eluent source through a first eluent chromatography flow path, thereby performing elution in the chromatography apparatus, and operating the second pump to provide a cleaning liquid to a first waste liquid container from a cleaning source through a second cleaning waste liquid flow path, thereby at least partially simultaneously priming at least the second pump with the cleaning liquid; - Operating the second pump to provide a cleaning liquid to the chromatography apparatus from a cleaning source through a second cleaning chromatography flow path, thereby re-equilibrating the chromatography apparatus, and operating the first pump to provide a cleaning liquid to a first waste container from a cleaning source through a first cleaning waste flow path, thereby at least partially simultaneously priming at least the first pump with the cleaning liquid; It is configured to operate by performing the above.

[0021] According to one embodiment, the preparative chromatography system is configured such that during the step of performing elution, the eluate from the chromatography apparatus is directed by a lower valve to a fraction collector or a second waste container based on the content of the eluate determined from the signal output from the monitoring unit.

[0022] According to one embodiment, the preparative chromatography system includes an upper monitoring unit configured in front of the flow direction to the first waste container, and the upper monitoring unit is configured to analyze the content in the first waste flow path and the second waste flow path.

[0023] Thanks to the present invention, the hold-up volume is reduced, which facilitates the use of small-volume chromatography devices in preparative chromatography systems. This is particularly important in preparative chromatography processes that utilize a large number of processing cycles. Compared to prior art preparative chromatography systems, the hold-up volume is reduced by a factor of 10 to 100. As a result, the resolution is improved, and the broadening of chromatography peaks, i.e., dispersion, can be significantly reduced. Furthermore, the consumption of the processing liquid is significantly reduced. Similarly, the processing time is significantly shortened.

[0024] One advantage of the present invention is that a preparative chromatography system may include a plurality of chromatography devices, and the advantageous low hold-up volume is maintained.

[0025] A further advantage is that the above methods and systems can be adapted to utilize more processing liquids, such as, for example, a plurality of different washing (buffer) liquids.

[0026] A further advantage of the present invention is that the system configuration provides a high degree of flexibility in designing the processing sequence and high efficiency in using the system.

[0027] Hereinafter, with reference to the accompanying drawings, the present invention will be described in more detail by way of non-limiting examples with respect to its embodiments.

Brief Description of the Drawings

[0028]

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[0029] The following description is for the purpose of illustration and explanation of the present invention only and is not intended to limit the present invention to the specific embodiments described.

[0030] Unless otherwise defined, technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0031] All references cited herein, including patent applications and publications, are incorporated by reference in their entirety.

[0032] Terms such as "upper", "upstream", "lower", and "downstream" are used herein in relation to the general liquid flow direction of a chromatography system and indicate the order of components and units of the system and are in no way intended to limit the present invention. The "upper" portion refers to the portion of the flow path of the chromatography system that is upstream of the central unit, i.e., the chromatography apparatus or apparatuses. Thus, "lower" refers to the portion downstream of the chromatography apparatus.

[0033] The systems and methods according to the present invention are described, in their different embodiments, using three basic processing fluids: an eluent, a cleaning fluid, and a supply fluid. This is intended as an exemplary example representing a minimum set of processing fluids, and those skilled in the art will understand that more and / or other processing fluids can be used and are often used. For example, typically many different buffers are used for cleaning and equilibrating chromatography devices. Also, a controlled change in the mobile phase composition during chromatography runs, so-called gradient chromatography, can be combined with the methods and systems according to embodiments of the present invention by adaptations obvious to those skilled in the art.

[0034] To explain the configuration and settings of the described preparative chromatography system, a certain degree of functional attenuation is used. The term "connected", such as "pump A connected to valve B", should be understood to mean that fluid communication is provided between two units, for example by a pipe or a channel. The phrase "set valve A to connect source Ba to container C" should be understood to mean that valve A has and is set to provide fluid communication between source B and container C. Such functional attenuation is only for the sake of brevity, and those skilled in the art should have no problem providing the configuration or the correct settings. Functional attenuation is also used with respect to specific parts of the system, such as valves. For example, a four-way valve can be realized using parts of a multi-way valve, or by combining, for example, a two-way valve and a three-way valve. Similarly, the function of a check valve may be provided by a control valve.

[0035] To facilitate understanding of the present invention, a prior art preparative chromatography system will be briefly described with reference to the schematic diagram of FIG. 1. The prior art preparative chromatography system 100 includes a source 101a, an eluent source 101b, and a cleaning liquid source 102b for providing a processing liquid, a supply liquid, an eluent, and a cleaning liquid. The source 101a, the eluent source 101b, and the cleaning source 102b are all connected to a common first valve 103 via appropriate pipe devices, and the valve 103 is connected to a common pump 104 via a first common pipe device 111. The common pump 104 is further connected to a chromatography device 105, such as a conventional packed bed column, via a second common pipe device 112. The first valve is arranged to direct the flow from any of the three sources to a common valve outlet. The chromatography device 105 is connected to a monitoring device 106 in relation to a lower valve 107 arranged to direct the flow to either a fraction collector 108 or a waste container 109. As shown in FIG. 1, the prior art system has a common flow path, indicated by a dotted line, extending from the common first valve 103, through the first common piping configuration 110, the common pump 104, and through the second common piping configuration to the chromatography device 105. The length of the common flow path and the number and type of units (e.g., pumps and valves) are directly related to the hold-up volume of the system 100, as will be recognized by those skilled in the art. It should be noted that not only is the volume formed within the pipes important, but the volume contained within valves, particularly more complex valves having multiple inlets and / or outlets, is typically a significant amount. However, in the illustrated prior art system 100, the hold-up volume due to a reasonably skilled selection of units and pipes is small compared to the volume of a conventional column containing a porous bead-based chromatography medium and is not a major concern regarding the overall performance of the system.

[0036] A preparative chromatography system 200 for chromatographic separation of biological or chemical samples according to the present invention is schematically shown in FIG. 2a. The preparative chromatography system 200 includes a source 201a, a cleaning source 201b and an eluent source 201c for providing a processing solution, a feed, a cleaning agent and an eluent. The source 201a is connected to the chromatography device 205 via a first supply pipe device 210a, a supply pump 203a, a second supply pipe device 211a, an upper supply valve 204a and a third supply pipe device 212a. The cleaning source 202b is connected to the chromatography device 205 via a first cleaning pipe arrangement 210b, a cleaning pump 203b, a second cleaning pipe arrangement 211b, an upper cleaning valve 204b and a third cleaning pipe arrangement 212b. The eluent source 201c is connected to the chromatography device 205 via a first eluent pipe arrangement 210c, an eluent pump 203c, a second eluent pipe arrangement 211c, an upper eluent valve 204c and a third eluent pipe arrangement 212c. The chromatography device 205 is preferably a low-volume device such as a membrane chromatography device, and even more preferably a fiber-based membrane chromatography device. The chromatography device 205 is connected to a monitoring unit 206 in relation to a lower valve 207 arranged to direct the flow to either a fraction collector 208 or a waste container 209. The monitoring unit 206 can be provided with a combination of detectors, for example, one detector, such as a UV detector, a photo diode array (PDA) detector, or a UV detector combined with a mass spectrometer (MS). As will be understood by those skilled in the art, a number of fraction collectors can be utilized, and also a number of separate waste containers can be utilized.

[0037] The fractionation chromatography system 200 further includes a control unit (not shown) communicatively connected to at least pumps 203a, 203b, 203c and a lower valve 207, which is typically a three-way control valve. When control valves are used, upper valves 204a, 204b, 204c are also connected to the control unit and can be controlled by the control unit. The control unit and the valves controlled by the control unit are also provided in other embodiments presented below.

[0038] As shown in FIG. 2b, the fractionation chromatography system 200 according to the present invention provides separate individual upper flow paths from each process liquid source to or near the chromatography device 205. Each individual upper flow path ends at a mixing point 221 defined as the first position in the flow direction, where the process liquids meet and are mixed with another process liquid. The supply upstream flow path 220a is shown by a dotted line, the cleaning upstream flow path 220b is shown by a dashed line, and the eluent upstream flow path 220c is shown by a chain double-dashed line. In FIG. 2a, the mixing point is at the upper part of the chromatography device 205. For example, if the supply upper flow path merges with the cleaning flow path upstream of where it merges with the eluent upper flow path, a system with multiple mixing points can be envisioned. By introducing separate pumps and valves into each upper flow path, the length of the common flow path is minimized, and in the embodiment shown in FIG. 2a, the common flow path is limited to the upper part of the chromatography device 205, and the process liquids are first mixed when changing from one process liquid to another. This can be compared with the longer flow paths discussed with reference to the prior art system of FIG. 1. Therefore, with the novel design, the hold-up volume can be significantly reduced. The novel design of the fractionation chromatography system 200 facilitates a standby mode in which the supply flow path, the eluent flow path, and the cleaning flow path are each filled with their respective process liquids. Therefore, during the processing cycle, when shifting from one process liquid, the next process liquid is always immediately available, minimizing unwanted mixing of the process liquids. This will be further discussed when explaining the operating method according to the present invention.

[0039] The chromatography device 205 is shown in FIG. 2a with a plurality of inlets, a supply inlet 213a, a cleaning inlet 213b, and an eluent inlet 213c. To control the flow of liquid within the chromatography device 205, an internal manifold 214a can be provided at the inlet portion of the chromatography device 205, which is an embodiment of the present invention schematically shown in FIG. 2c (only the relevant parts of the system are shown). The internal manifold 214a provides fluid communication from the supply inlet 213a, the cleaning inlet 213b, and the eluent inlet 213c to a common manifold outlet 215a arranged to supply the processing liquid to the active part of the chromatography device 205, i.e., the part that holds the chromatography medium. The internal manifold 214a ensures that all the processing liquids enter the active part of the chromatography device 205 at the same position. In this embodiment, the supply flow path shown by the dotted line, the eluent flow path shown by the dashed line, and the cleaning flow path shown by the chain double-dashed line extend from their respective sources to the branch point of the internal manifold 214b.

[0040] According to one embodiment schematically shown in FIG. 2d, the chromatography device 205 has a single inlet 213, and the system 200 includes an external manifold 214b that provides fluid communication from the supply inlet 213d, the wash inlet 213e, and the eluent inlet 213f to a common external manifold outlet 215b. The common external manifold outlet 215b is connected to the single inlet 213 of the chromatography device 205. In this embodiment, the supply flow path shown by the dotted line, the eluent flow path shown by the dashed line, and the wash flow path shown by the two-dot chain line each extend from their respective sources to the branch points of the external manifold 214b. Preferably, according to one embodiment of the present invention, each of the upper valves 204a, 204b, 204c is disposed near each inlet of the external manifold 214b to form a valve manifold assembly 214c. "Closing" should be understood as disposing the upper valve as close as possible to the external manifold 214b in consideration of the details of the structures of different objects. Functionally, the valve manifold assembly 214c can be regarded as a manifold equipped with an inlet valve that allows only downstream flow, for example, a check valve, so that a certain processing liquid cannot enter the upstream flow path of another processing liquid.

[0041] According to one embodiment schematically shown in FIG. 2e, the valve manifold assembly is realized by a multi-way valve 214d having three inlets and one outlet, for example, a four-way valve. For each valve position, the multi-way valve 214d opens between one of the inlets and the outlet, leaving the remaining inlets closed.

[0042] According to an embodiment of the present invention, the fractionation chromatography system 200 includes a purge device 216 schematically shown in FIGS. 2f-2g. According to one embodiment shown in FIG. 2f, fluid communication is preferably constituted by a piping device 217 between a purge outlet 218 of the chromatography device 205 and a waste container 209, provided in the vicinity of an inlet 213 or inlets 213a, 213b, 213c of the chromatography device 205. The purge valve 218 is a control valve connected to and controlled by a control unit, and regulates the flow of the processing liquid from the chromatography device 205 to the waste container 209.

[0043] In an alternative embodiment of the purge device 216 schematically shown in FIG. 2g, an external manifold 214b or a valve manifold assembly 214c is provided, and a purge valve 218b is provided between and connected to an external manifold outlet 215b and a chromatography device inlet 213. In this embodiment, the purge valve 218b is preferably a three-way control valve arranged to provide fluid communication to either the chromatography device 205 or the waste container 209, and is connected to and controlled by a control unit.

[0044] A further alternative embodiment of the purge device 216 is that purge valves are provided in each of a supply flow path 220a, a washing flow path 220b, and an eluent flow path 220c. This can be achieved by providing the upper valves 203a, 203b, 203c as three-way valves arranged to provide fluid communication to either the chromatography device 205 or the waste container 209, or to be in a closed position.

[0045] According to one embodiment of the present invention schematically shown in FIG. 2h, at least in the eluent flow path, means for circulating the eluent in the loop configuration 220 is provided. The loop configuration 220 includes an eluent pump 203c, a loop control valve 221, and a piping arrangement that connects a first eluent piping arrangement 210c upstream of the eluent pump 203c and a second eluent piping arrangement 211c downstream. A valve 220, preferably a control valve, is provided upstream of the connection between the loop piping and the first eluent piping configuration 210c. Alternatively, the connection of the first eluent pipe arrangement 210c of the loop pipe may be by a three-way valve. The loop configuration 220 provides continuous circulation of the eluent in the loop upstream of the upper eluent valve 204c, and the eluent pump 203b can operate continuously. The loop configuration 220 reduces the negative impact of the startup time and otherwise contributes to the hold-up time / volume of the system when shifting between different processing liquids. Also, corresponding loop configurations can be provided in the supply and washing flow paths.

[0046] The method of the present invention for operating a preparative chromatography system will be described with reference to the flow path diagrams of FIGS. 3a to 3h and the flowchart of FIG. 4. As an exemplary but non-limiting example of a preparative chromatography system suitable for carrying out the method of the present invention, reference is made to the preparative chromatography system 200 described with reference to FIGS. 2a to 2f. FIGS. 3a to 3h include an upper portion showing a preparative chromatography system having different settings for carrying out the steps of the method, and FIGS. 3d to 3g also include a lower portion which is a schematic graph showing the output from the monitoring unit 206, with the solid line showing the output related to the current step and the dashed line showing the output related to the previous step.

[0047] The method need not necessarily be carried out in the following order, but includes a preliminary step 400 with the preliminary steps shown in FIGS. 3a to 3c. 400:1: Prime the upper supply flow path 220a with the supply liquid provided from the supply source 201. According to one embodiment, control the supply pump 203a and the upper supply valve 204a to load the supply liquid into the supply flow path 220a, and close the upper cleaning valve 204b and the upper eluent valve 204c during the supply priming period. 400:2: Prime the upper cleaning flow path 220b with the cleaning liquid provided from the cleaning source 201b. According to one embodiment, control the cleaning pump 203b and the upper cleaning valve 204b to load the cleaning liquid into the cleaning flow path 220b, and close the upper supply valve 204a and the upper eluent valve 204c during this cleaning priming period. 400:3: Prime the upper eluent flow path 220c with the eluent provided from the eluent source 201c. According to one embodiment, control the eluent pump 203c and the upper eluent valve 204c to load the eluent into the eluent flow path 220c, and close the upper cleaning valve 204b and the upper supply valve 204a during this eluent priming period.

[0048] The previous step 400 thereby provides a standby mode with the processing liquid loaded up to at least the mixing point.

[0049] Following the previous step, the preparative chromatography system 200 transitions to the active preparative stage and performs the actual separation process (steps 410 to 450). The separation process is repeated or cycled a predetermined number of times, for a predetermined period, or until a predetermined criterion is met, for example, until a predetermined fraction volume is collected or until all the supply liquid is consumed. The order of the steps, the duration of the steps, and other processing parameters are stored in the processing file and executed by the control unit. The steps to be taken consecutively are 410: By providing the cleaning liquid from the cleaning source 201b to the chromatography device 205 via the upper cleaning flow path 220b, the chromatography device is equilibrated. According to one embodiment, by controlling the cleaning pump 203b and the upper cleaning valve 204b to load the cleaning liquid into the chromatography device 205 and through it, the lower valve 207 is set to the waste container 209, and during this step, the upper supply valve 204a and the upper eluent valve 204c are closed. 420: By providing the supply liquid from the supply source 201a to the chromatography device 205 via the upper supply flow path 220a, the supply liquid is loaded into the chromatography device. According to one embodiment, by controlling the supply pump 203a and the upper supply valve 204a to load the supply liquid into the chromatography device 205 and through it, the lower valve 207 is set to the waste container 209, and during this step, the upper cleaning valve 204b and the upper eluent valve 204c are closed. 430: By providing the cleaning liquid from the cleaning source 201b to the chromatography device 205 via the upper cleaning flow path 220b, the chromatography device is cleaned. According to one embodiment, by controlling the cleaning pump 203b and the upper cleaning valve 204b to load the cleaning liquid into the chromatography device 205 and through it, the lower valve 207 is set to the waste container 209, and during this step, the upper supply valve 204a and the upper eluent valve 204c are closed. 440: By providing the eluent provided from the eluent source 201c to the chromatography device 205 via the upper eluent flow path 220c, elution is performed within the chromatography device. According to one embodiment, by controlling the eluent pump 203c and the upper eluent valve 204c to load the eluent into the chromatography device 205 and through it, the lower valve 207 is set to either the waste container 209 or the fraction collector 208, and the setting depends on the content of the eluate detected by the monitoring unit 206. During this step, the upper cleaning valve 204b and the upper supply valve 204a are closed. 450: By providing the cleaning liquid from the cleaning source 201b to the chromatography device 205 through the upper cleaning flow path 220b, the chromatography device is re-equilibrated. According to one embodiment, the cleaning pump 203b and the upper cleaning valve 204b are controlled to load the cleaning liquid into and through the chromatography device 205, with the lower valve 207 set to the waste container 209, and during this step, the upper supply valve 204a and the upper eluent valve 204c are closed.

[0050] After the first cycle, the equilibration step 410 may be optional or combined with the re-equilibration step 450.

[0051] The duration of each step can preferably be dynamically determined using a so-called monitoring function that utilizes the output from the monitoring unit 206 to determine that the processing of the chromatography device 205, which is the purpose of the step, has achieved the desired effect. Alternatively, for certain steps that utilize the monitoring function, while for other steps, for example, when they are not so important for performance, the duration is a time-based parameter.

[0052] According to one embodiment of this method, one or more of the sub-steps of the previous stage include purging the individual upper flow paths by controlling the purge valve to open and direct the flow of the processing liquid to the waste container 209.

[0053] According to one embodiment, at least the upper eluent flow path 220c is provided with a loop configuration 220, and during the steps of the active separation stage, the eluent pump 203c is always controlled to be active, and when the upper eluent valve 204c is closed, that is, in all steps except the separation step of loading the eluent into the chromatography device 205, the loop control valve 221 is controlled to open.

[0054] One embodiment of the present invention schematically shown in FIGS. 5a to 5b is a preparative chromatography system 500 including, but not limited to, a plurality of chromatography devices, for example, a first chromatography device 205a, a second chromatography device 205b, and a third chromatography device 205c. Shown in FIG. 5a is the main flow path of the preparative chromatography system 500.

[0055] The preparative chromatography system 200 includes a supply source 201a, a cleaning source 202b, and an eluent source 201c for providing a processing liquid, a feed, a cleaning agent, and an eluent. Associated pumps, a supply pump 203a, a cleaning pump 203b, and an eluent pump 203c are arranged at each supply source as in the above-described embodiment. Downstream of the pumps, individual flow paths branch due to the arrangement of pipes and valves, and as a result, the following occurs.

[0056] A first upper supply flow path 220a-1 is provided from the supply source 201a to the first chromatography device 205a, a second upper supply flow path 220a-2 is provided from the supply source 201a to the second chromatography device 205b, and a third upper supply flow path 220a-3 is provided from the supply source 201a to the third chromatography device 205c indicated by a dotted line.

[0057] A first upper cleaning flow path 220b-1 is provided from the cleaning source 201b to the first chromatography device 205a, a second upper cleaning flow path 220b-2 is provided from the cleaning source 201a to the second chromatography device 205b, and a third upper cleaning flow path 220a-3 is provided from the cleaning source 201a to the third chromatography device 205c indicated by a broken line.

[0058] A first upper eluent flow path 220c-1 is provided from the eluent source 201c to the first chromatography device 205a, a second upper eluent flow path 220b-2 is provided from the eluent source 201a to the second chromatography device 205b, and a third upper eluent flow path 220c-3 indicated by a broken line is provided from the eluent source 201a to the third chromatography device 205c.

[0059] The preparative chromatography system 200 can operate according to a parallel hierarchy, for example, compared with a conventional system. For example, in various embodiments, a cleaning liquid can be provided to each of the chromatography devices 205a, 205b, 205c substantially simultaneously via the flow paths 220b-1, 220b-2, and 220b-3.

[0060] One monitoring unit 206a, 206b, 206c is provided downstream of the chromatography device and is connected to each chromatography device 205a, 205b, 205c. Through the arrangement of pipes and valves, the chromatography devices 205a, 205b, 205c are connected to either a common waste container 209 and a common fraction collector 208 (Figure 5a), or waste containers 209a, 209b, 209c are separated for each of the chromatography devices 205a, 205b, 205c and the common fraction collector 208 (Figure 5b), or a waste container and a separate fraction collector are separated for each chromatography device (not shown).

[0061] The embodiment described with reference to Figures 2a to 2f relates to purging and providing means for circulating one or more of the processing liquids, and is also related to a preparative chromatography system 500 that provides a plurality of chromatography devices.

[0062] According to one embodiment schematically shown in FIG. 5b, individual flow paths are realized by the arrangement of piping and four-way valves. In the following description, the term "connected" is used to indicate that a unit such as a four-way valve is fluidly connected to, for example, a chromatography device via an appropriate pipe, and the details of the pipe are not described in detail. The supply pump 203a is connected to the first four-way valve 501, the cleaning pump 203b is connected to the second four-way valve 502, and the eluent pump 203c is connected to the third four-way valve 503. The first four-way valve 501, the second four-way valve 502, and the third four-way valve 503 can be regarded as a first-level processing liquid distribution unit. The first four-way valve 501 is connected to the fourth four-way valve 504, the fifth four-way valve 505, and the sixth four-way valve 506. The second four-way valve 502 is connected to the fourth four-way valve 504, the fifth four-way valve 505, and the sixth four-way valve 506. The third four-way valve 502 is connected to the fourth four-way valve 504, the fifth four-way valve 505, and the sixth four-way valve 506. The fourth four-way valve 504 is connected to the first chromatography device 205a, the fifth four-way valve 505 is connected to the second chromatography device 205b, and the sixth four-way valve 506 is connected to the third chromatography device 205c. The fourth four-way valve 504, the fifth four-way valve 505, and the sixth four-way valve 506 can be regarded as a second-level processing liquid distribution unit, and the mixing point is defined as above as the first position in the flow path where mixing of different processing liquids occurs, and is at the fourth four-way valve 504, the fifth four-way valve 505, and the sixth four-way valve 506. That is, the mixing point is at the same level, the second level. The four-way valves are arranged such that when they are open from the inlet to the first outlet / connection, the remaining two outlets / connections are closed. Thereby, individual flow paths from each supply source to each chromatography device can be provided. For example, the supply source 201a is connected to the first four-way valve 501 via the supply pump 203a, and the first four-way valve 501 provides a flow path to the fourth four-way valve 504 at a certain position, and then, at a certain position, opens the inlet connected to the first four-way valve 501 and provides a flow path to the first chromatography device 205a.The second position of the first four-way valve 501 provides a flow path to the fifth four-way valve 504 and then, at a certain position, opens the inlet connected to the first four-way valve 501 to provide a flow path to the second chromatography device 205b.

[0063] One embodiment of the method according to the present invention can utilize a preparative chromatography system 500 equipped with a plurality of chromatography devices and will be described with reference to the flow path diagrams of FIGS. 6a - 6b and the flowchart of FIG. 7. As an exemplary but non-limiting example of a preparative chromatography system suitable for performing the method according to this embodiment, reference is made to the preparative chromatography system 500 described with reference to FIGS. 5a - 5b.

[0064] FIGS. 6a - 6b are examples of how the preparative chromatography system 500 can be operated, a) showing the system in standby mode, b) showing elution in the first chromatography device 205a and equilibration in the third chromatography device 205c, c) showing the cleaning of the first chromatography device 205a, d) showing the loading of the feed to the first chromatography device 205a, e) showing the loading of the feed solution to the first chromatography device 205a and the equilibration of the second chromatography device 205b, f) showing the cleaning of the first chromatography device 205a and the loading of the feed to the second chromatography device 205b, g) showing the loading of the eluate to the first chromatography device 205a, the loading of the feed solution to the second chromatography device 205b, and the equilibration of the third chromatography device 205c, h) showing the retention of the first chromatography device 205a, the cleaning of the second chromatography device 205b, and the loading of the feed solution to the third chromatography device 205c. i) Shows the equilibration of the first chromatography device 205a, the loading of the eluate into the second chromatography device 205b, and the loading of the feed solution into the third chromatography device 205c. j) Shows the loading of the feed solution into the first chromatography device 205a, the retention of the second chromatography device 205b, and the cleaning of the third chromatography device 205c. k) Shows the loading of the feed into the supply device 1 of the first chromatography device 205a, the cleaning of the second chromatography device 205b, and the loading of the eluate into the third chromatography device 205c.

[0065] Each separate chromatography device 505a - c experiences a sequence of processing liquids as shown in FIGS. 3c - 3h. For example, chromatography device 505a is one step ahead of chromatography device 505b, and chromatography device 505b is one step ahead of chromatography device 505c.

[0066] This method does not necessarily have to be executed in the following order, but includes a previous step 700 with the following steps. 700:1: Priming the first upper supply flow path 220a - 1 associated with the first chromatography device 205a with the feed solution provided from the supply source 201a, priming the second upper supply flow path 220a - 2 associated with the second chromatography device 205b with the feed solution provided from the supply source 201a, priming the third upper supply flow path 220a - 3 associated with the third chromatography device 205c with the feed solution provided from the supply source 201a, and priming with the feed solution engaging the supply pump 203a. 700:2: Priming the first upper cleaning channel 220b-1 associated with the first chromatography device 205a with the cleaning liquid provided from the cleaning source 201b, priming the second upper cleaning channel 220b-2 associated with the second chromatography device 205b with the cleaning liquid provided from the cleaning source 201a, priming the third upper cleaning channel 220a-3 associated with the third chromatography device 205c with the cleaning liquid provided from the cleaning source 201a, and priming with the cleaning liquid engaged with the cleaning pump 203b. 700:3: Priming the first upper eluent channel 220c-1 associated with the first chromatography device 205a with the eluent provided from the eluent source 201c, priming the second upper eluent channel 220c-2 associated with the second chromatography device 205b with the eluent provided from the eluent source 201c, priming the third upper eluent channel 220c-3 associated with the third chromatography device 205c with the eluent provided from the eluent source 201c, and priming with the eluent engaged with the eluent pump 203c.

[0067] The previous step 700 thereby provides a standby mode with the processing liquid loaded up to at least the mixing point associated with each chromatography device. The priming of the upper channels during the previous step is preferably performed simultaneously or at least partially with temporal overlap so that the respective processing fluids are loaded into the first upper supply channel 220a-1, the first upper cleaning channel 220b-1, and the first upper eluent channel 220c-1 at the same time.

[0068] Subsequent to the previous stage, the preparative chromatography system 500 proceeds to the active preparative stage and performs the actual separation process (Steps 710 - 730). The separation process is repeated or cycled a predetermined number of times, for a predetermined period, or until a predetermined criterion is met, for example, until a predetermined fraction volume is collected. The order of steps, the duration of steps, and other processing parameters are stored in a processing file, executed by the control unit, and the sub - steps of the main steps are at least partially temporally overlapping, 710:1 Step of equilibrating or washing the first chromatography device 205a with the cleaning liquid provided from the cleaning source 201b through the first upper cleaning flow path 220b - 1 by operating the cleaning pump 203b during the next step 710:2 Step of loading the supply liquid provided from the supply source 201a through the second upper supply flow path 220a - 2 into the second chromatography device 205b by operating the supply pump 203a, 710:3 Step of performing elution in the third chromatography device 205c by providing an eluent from the eluent source 201c through the third upper eluent flow path 220c - 3 by operating the eluent pump 203c, wherein the eluent is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206, or 710:2’ Step of loading the supply liquid provided from the supply source 201a through the third upper supply flow path 220a - 3 into the third chromatography device 205c by operating the supply pump 203a, 710:3’ Step of performing elution in the second chromatography device 205c by providing an eluent from the eluent source 201c through the second upper eluent flow path 220c - 2 by operating the eluent pump 203c, wherein the eluent is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206, at least one of which is executed 720:1 By operating the cleaning pump 203b, a step of equilibrating or cleaning the second chromatography device 205b with the cleaning liquid provided from the cleaning source 201b through the second upper cleaning flow path 220b-2 during the next step 720:2 By operating the supply pump 203a, a step of loading the supply liquid provided from the supply source 201 through the first upper supply flow path 220a-1 into the first chromatography device 205a 720:3 By operating the eluent pump 203c to provide an eluent from the eluent source 201c through the third upper eluent flow path 220c-3, a step of performing elution in the third chromatography device 205c, wherein the eluent is directed to a waste container or a fraction collector according to the content of the eluate detected by the monitoring unit 206 or 720:2’ By operating the supply pump 203a, a step of loading the supply liquid provided from the supply source 201 through the third upper supply flow path 220a-3 into the third chromatography device 205c 720:3’ By operating the eluent pump 203c to provide an eluent from the eluent source 201c through the first upper eluent flow path 220c-1, a step of performing elution in the first chromatography device 205a, wherein the eluent is directed to a waste container or a fraction collector according to the content of the eluate detected by the monitoring unit 206 at least one of which is executed 730:1 By operating the cleaning pump 203b, a step of equilibrating or cleaning the third chromatography device 205c with the cleaning liquid provided from the cleaning source 201b through the third upper cleaning flow path 220b-3 during the next step Step of loading the supply liquid provided from the supply source 201 through the first upper supply channel 220a-1 into the first chromatography device 205a by operating the supply pump 203a, Step of performing elution with the second chromatography device 205b by providing an eluent from the eluent source 201c through the second upper eluent channel 220c-2 by operating the eluent pump 203c, wherein the eluent is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206, Or, Step of loading the supply liquid provided from the supply source 201 through the second upper supply channel 220a-2 into the second chromatography device 205b by operating the supply pump 203a, Step of performing elution with the first chromatography device 205a by providing an eluent from the eluent source 201c through the first upper eluent channel 220c-1 by operating the eluent pump 203c, wherein the eluent is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206, At least one of them is executed.

[0069] The four-way valve should be interpreted as a valve or valve arrangement that provides at least one inlet and at least three outlets by changing the valve setting / position, and provides fluid communication between the inlet and any one of the three outlets.

[0070] According to an embodiment of the present invention, the steps of the previous stage 700 are, 700b:1: By setting the first four-way valve 501 to direct the flow towards the fourth four-way valve 504 and setting the fourth four-way valve 504 to direct the flow towards the first chromatography device 205a, prime the first upper supply channel 220a-1 related to the first chromatography device 205a with the supply liquid provided from the supply source 201, Set the first four-way valve 501 to direct the flow towards the fifth four-way valve 505, and set the fifth four-way valve 505 to direct the flow towards the second chromatography device 205b, thereby priming the second upper supply channel 220a-2 related to the second chromatography device 205b with the supply liquid provided from the supply source 201a. A step of setting the first four-way valve 501 to direct the flow towards the sixth four-way valve 506, and setting the sixth four-way valve 506 to direct the flow towards the third chromatography device 205c, thereby priming the third upper supply channel 220a-3 related to the third chromatography device 205c with the supply liquid provided from the supply source 201a. 700b:2: Set the second four-way valve 502 to direct the flow towards the fourth four-way valve 504, and set the fourth four-way valve 504 to direct the flow towards the first chromatography device 205a, thereby priming the first upper cleaning channel 220b-1 related to the first chromatography device 205a with the cleaning liquid provided from the cleaning source 201b. Set the second four-way valve 502 to direct the flow towards the fifth four-way valve 505, and set the fifth four-way valve 505 to direct the flow towards the second chromatography device 205b, thereby priming the second upper cleaning channel 220b-2 related to the second chromatography device 205b with the cleaning liquid provided from the cleaning source 201a. A step of setting the second four-way valve 502 to direct the flow towards the sixth four-way valve 506, and setting the sixth four-way valve 506 to direct the flow towards the third chromatography device 205c, thereby priming the third upper cleaning channel 220a-3 related to the third chromatography device 205c with the cleaning liquid provided from the cleaning source 201a. 700b:3: Set the third four-way valve 503 to direct the flow towards the fourth four-way valve 504, and set the fourth four-way valve 504 to direct the flow towards the first chromatography device 205a, thereby priming the first upper eluent channel 220c-1 related to the first chromatography device 205a with the eluent provided from the eluent source 201c. Set the third four-way valve 503 to direct the flow towards the fifth four-way valve 505, and set the fifth four-way valve 505 to direct the flow towards the second chromatography device 205b, thereby priming the second upper eluent flow path 220c-2 related to the second chromatography device 205b with the eluent provided from the eluent source 201c. A step of setting the third four-way valve 503 to direct the flow towards the sixth four-way valve 506, and setting the sixth four-way valve 506 to direct the flow towards the third chromatography device 205c, thereby priming the third upper eluent flow path 220c-3 related to the third chromatography device 205c with the eluent provided from the eluent source 201c. It is executed according to

[0071] In this embodiment, the steps of the active separation stage are executed according to steps 710b to 730b. 710b:1 A step of setting the second four-way valve 502 to direct the flow towards the fourth four-way valve 504, and setting the fourth four-way valve 504 to direct the flow towards the first chromatography device 205a, thereby equilibrating or cleaning the first chromatography device 205a with the cleaning liquid provided from the cleaning source 201b through the first upper cleaning flow path 220b-1. During 710b:2 A step of setting the first four-way valve 501 to direct the flow towards the fifth four-way valve 505, and setting the fifth four-way valve 505 to direct the flow towards the second chromatography device 205b, thereby loading the supply liquid provided from the supply source 201a into the second chromatography device 205b through the second upper supply flow path 220a-2. 710b:3 A step of setting the third four-way valve 503 to direct the flow towards the sixth four-way valve 506, and setting the sixth four-way valve 506 to direct the flow towards the third chromatography device 205c, thereby performing elution in the third chromatography device 205c by providing the eluent from the eluent source 201c through the third upper eluent flow path 220c-3, wherein the eluate is directed towards the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206. or 710b:2’ By setting the first four-way valve 501 to direct the flow toward the sixth four-way valve 506 and setting the sixth four-way valve 506 to direct the flow toward the third chromatography device 205c, the supply liquid provided from the supply source 201 through the third upper supply channel 220a-3 is loaded into the third chromatography device 205c, 710b:3’ By setting the third four-way valve 503 to direct the flow toward the fifth four-way valve 505 and setting the fifth four-way valve 505 to direct the flow toward the second chromatography device 205b, and by providing the eluent from the eluent source 201c through the second upper eluent channel 220c-2, the step of performing elution in the second chromatography device 205b, wherein the eluate is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206, at least one of which is executed, 720b:1 By setting the second four-way valve 502 to direct the flow toward the fifth four-way valve 505 and setting the fifth four-way valve 505 to direct the flow toward the second chromatography device 205b, the step of equilibrating or cleaning the second chromatography device 205b with the cleaning liquid provided from the cleaning source 201b through the second upper cleaning channel 220b-2, during the following step 720b:2 By setting the first four-way valve 501 to direct the flow toward the fourth four-way valve 504 and setting the fourth four-way valve 504 to direct the flow toward the first chromatography device 205a, the step of loading the supply liquid provided from the supply source 201 through the first upper supply channel 220a-1 into the first chromatography device 205a, 720b:3 By providing the eluent from the eluent source 201c through the third upper eluent channel 220c-3, the step of performing elution in the third chromatography device 205c, wherein the eluate is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206, or 720b:2’ By setting the first four-way valve 501 to direct the flow towards the sixth four-way valve 506 and setting the sixth four-way valve 506 to direct the flow towards the third chromatography device 205c, the supply liquid provided from the supply source 201 through the third upper supply flow path 220a-3 is loaded into the third chromatography device 205c. 720b:3’ By setting the third four-way valve 503 to direct the flow towards the fourth four-way valve 504 and setting the fourth four-way valve 504 to direct the flow towards the first chromatography device 205a, the eluent is provided from the eluent source 201c through the first upper eluent flow path 220c-1, and elution is performed in the first chromatography device 205a. The eluate is directed towards the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206. At least one of them is executed. 730b:1 By setting the second four-way valve 502 to direct the flow towards the sixth four-way valve 506 and setting the sixth four-way valve 506 to direct the flow towards the third chromatography device 205c, the third chromatography device 205c is equilibrated or washed with the cleaning liquid provided from the cleaning source 201b through the third upper cleaning flow path 220b-3. During 730b:2 By setting the first four-way valve 501 to direct the flow towards the fourth four-way valve 504 and setting the fourth four-way valve 504 to direct the flow towards the first chromatography device 205a, the supply liquid provided from the supply source 201 through the first upper supply flow path 220a-1 is loaded into the first chromatography device 205a. 730b:3 By setting the second four-way valve 502 to direct the flow towards the fifth four-way valve 505 and setting the fifth four-way valve 505 to direct the flow towards the second chromatography device 205b, the eluent is provided from the eluent source 201c through the second upper eluent flow path 220c-2, and elution is performed in the second chromatography device 205b. The eluate is directed towards the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206. Or 730b:2’ By setting the first four-way valve 501 to direct the flow towards the fifth four-way valve 505 and setting the fifth four-way valve 505 to direct the flow towards the second chromatography device 205b, the supply liquid provided from the supply source 201 through the second upper supply flow path 220a-2 is loaded into the second chromatography device 205b, 730b:3’ By setting the third four-way valve 503 to direct the flow towards the fourth four-way valve 504 and setting the fourth four-way valve 504 to direct the flow towards the first chromatography device 205a, the eluent is provided from the eluent source 201c through the first upper eluent flow path 220c-1, and elution is performed in the first chromatography device 205c. The eluate is directed towards the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit 206, At least one of them is executed.

[0072] According to an embodiment of the present invention schematically shown in FIGS. 8a - 8b, a preparative chromatography system 800 is configured such that two pumps operate in parallel. The preparative chromatography system 800 includes a supply source 801a, a cleaning source 801b, and an eluent source 801c for providing a processing solution, a feed, a cleaning solution, and an eluent. The preparative chromatography system 800 further includes a first pump 803a and a second pump 803b arranged to operate in a parallel configuration, which will be further discussed below. The first pump 803a is connected to the cleaning source 801b via a piping configuration 811a:a and a cleaning source valve 804b:a, and is connected to the eluent source 801c via a piping configuration 811b:a and an eluent source valve 804c:a. The second pump 803b is connected to the cleaning source 801b via a first piping configuration 811a:b and a cleaning source valve 804b:b, and is connected to the supply source 801a via a first piping configuration 811b:b and a supply source valve 804c:b. The source valves 804a,b are typically control valves. Alternatively, the system 800 comprises two sets of processing fluid sources (not shown) each including a supply source, a cleaning source, and an eluent source, such that each of the first and second pumps is connected to a separate set. The first pump 803a and the second pump 803b are downstream via second piping configurations 812a, 812b respectively connected to a first upper valve 818a and a second upper valve 818b, and the first upper valve 818a and the second upper valve 818b are downstream respectively via third pipe devices 813a, 813b connected to a common valve 870, a four-way valve, and via fourth pipe devices 814a, 814bb connected to a first waste container 815. The first upper valve 818a and the second upper valve 818b are typically three-way valves (one inlet and two outlets) and can be set respectively to direct the flow either to the waste container 815 or to the common valve 870. The common valve 870 is connected to the chromatography device 805, is arranged to direct the flow from the first or second upper valves 818a - 818b towards the chromatography device 805, and is arranged adjacent to the other of the upper valves 818a - 818b.This prevents the processing fluid from entering the other of the currently unused piping configurations 813a, 813b to direct the flow towards the chromatography apparatus 805. Alternatively, a manifold is used instead of the common valve 870. However, such a configuration needs to be used only when backflow in a temporarily "unused" tube arrangement is allowed. Alternatively, a manifold with a check valve at the inlet can also be used. Alternatively, the functions of the first upper valve 818a, the second upper valve 818b, and the common valve 870 can be provided by one large multi-way valve appropriately arranged. The chromatography apparatus 805 is connected to a monitoring unit 806 connected to a lower valve 807 arranged to direct the flow to either the fraction collector 808 or the second waste container 809. The first and second waste containers may be the same.

[0073] As previously pointed out, the use of three processing fluids represents an exemplary example of the minimum configuration of the preparative chromatography system 800, but in a typical configuration, more processing fluids are used. A person skilled in the art will have no problem modifying the configuration to handle more processing fluids according to the design principles described in the above embodiments. Other modifications such as the provision of the aforementioned loop and gradient techniques, but not limited to these, can also be utilized in the preparative chromatography system 800.

[0074] According to one embodiment, the preparative chromatography system 800 includes a plurality of cleaning fluids or buffer solutions, and each of the first pump 803a and the second pump 803b is arranged to handle a plurality of cleaning fluids provided from different liquid sources. According to one embodiment, only one of the first pump 803a and the second pump 803b is connected to the supply liquid source 801a, and the other pump is connected to the elution source 801c.

[0075] Shown in Fig. 8b is a general embodiment according to the same concept described with reference to Fig. 8a. The preparative chromatography system 800 according to the embodiment provides separate individual upper flow paths from the process liquid source to the common valve 870 via the respective first pump 803a and second pump 803b. The individual upper flow paths end at a mixing point 821 defined as a first position in the flow direction where the process liquid passing through the first pump 803a meets and mixes with the process liquid passing through the second pump 803b. The mixing point 821 is located within the common valve 870. Further, the separate flow paths associated with the first pump 803a and the second pump 803b end at the first waste container 815. The first chromatography flow paths 820b, 820c, which are controlled by the first chromatography flow paths 820b, 820c and pass the first pump 803a from the respective process liquid source, washing source 801b, and eluent source 801c to the common valve 870, are indicated by dotted lines. The first waste flow paths 821b, 821c from the respective process liquid source, washing source 801b, and eluent source, which are controlled by the first pump 803a and pass to the first waste container 815, are indicated by dashed lines. The second chromatography flow paths 822a, 822b from the respective process liquid source, supply source 801a, and washing source 801b, which are controlled by the second pump 803b and pass to the common valve 870, are indicated by double-dashed lines. The second waste flow paths 823a, 823b from the respective process liquid source, supply source 801a, and washing source 801b, which are controlled by the second pump 803a and pass to the waste container 815, are indicated by double-dashed lines.

[0076] According to an embodiment of the present invention, the preparative chromatography system 800 is provided with one or more upper monitoring units 816 provided in the waste flow path between the first upper valve 818a, the second upper valve 818b, and the first waste container 815. The upper monitoring unit 816 may be configured to provide a monitoring function indicating that the flow path currently being primed with one process liquid is sufficiently primed and ready for a change to the next process liquid.

[0077] In an embodiment of the fractionation chromatography system 800, while the first pump 803a is engaged in providing the processing liquid to the chromatography device 805, the second pump 803b is engaged in providing the processing liquid to the waste container 815, thereby facilitating and operating to prime at least a part of the associated chromatography flow path and vice versa. According to one embodiment, a chromatography device sequence of the processing liquid passing through the chromatography device 805 is established, and from the chromatography device sequence, separate sequences of the first pump 803a and the second pump 803b and the flow paths associated therewith can be generated. According to the sequence, in one step, the first pump 803a provides a processing liquid, such as a cleaning liquid, to the chromatography device 805, and at least partially simultaneously, the second pump 803b primes its associated flow path with a processing liquid, such as a supply liquid, which is the next processing liquid provided to the chromatography device 805 according to the chromatography device sequence. In the next step, the second pump 803b provides the processing liquid (supply liquid according to the example) used for priming in the previous step to the chromatography device 805, and at least partially simultaneously, the first pump 803a primes the next processing liquid according to the chromatography device sequence. As in this example, the next processing liquid provided to the chromatography device is a cleaning liquid, which indicates that the individual pumps may continue to send the same processing liquid according to their individual sequences, while the processing liquid provided to the chromatography device 805 shifts according to the chromatography device sequence. This will be further explained when describing the operation method according to the embodiment of the present invention described below.

[0078] According to one embodiment, the fractionation chromatography system 800 includes two or more chromatography devices 805, and the common valve 870 is a multi-way valve.

[0079] One embodiment of the method according to the present invention can utilize a preparative chromatography system 800 having a tandem pump configuration and is described with reference to the flow path diagram of FIG. 9. As an exemplary but non-limiting example of a preparative chromatography system suitable for performing the method according to this embodiment, reference is made to the preparative chromatography system 800 described with reference to FIGS. 8a-8b.

[0080] Focusing on the flow through the chromatography device 805, the method results in a sequence including equilibration, feed loading, washing, elution, and re-equilibration, as in the above-described embodiments. This separation process is repeated or cycled a predetermined number of times, for a predetermined period, or until a predetermined criterion is met, for example, until a predetermined fraction volume is collected. The order of steps, the duration of steps, and other processing parameters are stored in a processing file and executed by a control unit.

[0081] The method according to the embodiment includes the following steps, namely, 910: A step of equilibrating the chromatography device 805 by operating the first pump 803a to provide a cleaning liquid from the cleaning source 801b to the chromatography device 805 via the first cleaning chromatography flow path 820b, and at least partially simultaneously priming at least the second pump 803b with a supply liquid by operating the second pump 803b to provide a supply liquid from the supply source 801a to the first waste container 815 via the second supply waste flow path 823a. 920: A step of loading the feed into the chromatography device 805 by operating the second pump 803b to provide a supply liquid from the supply source 801a to the chromatography device 805 via the second supply chromatography flow path 822a, and at least partially simultaneously priming at least the first pump 803a with a cleaning liquid by operating the first pump 803a to provide a cleaning liquid from the cleaning source 801b to the first waste liquid container 815 via the first cleaning waste liquid flow path 821b. 930: A step of cleaning the chromatography device 805 by operating the first pump 803a to supply the cleaning liquid from the cleaning source 801b to the chromatography device 805 via the first cleaning chromatography flow path 820b, and operating the second pump 803b to supply the cleaning liquid from the cleaning source 801b to the first waste liquid container 815 via the second cleaning waste liquid flow path 823b, thereby at least partially simultaneously priming at least the second pump 803b with the cleaning liquid. 940: A step of continuing to clean the chromatography device 805 by operating the second pump 803b to supply the cleaning liquid from the cleaning source 801b to the chromatography device 805 via the second cleaning chromatography flow path 821b, and operating the first pump 803a to supply the eluent liquid from the eluent source 801c to the first waste container 815 via the first eluent waste flow path 821c, thereby at least partially simultaneously priming at least the first pump 803a with the eluent liquid. 950: A step of performing elution in the chromatography device 805 by operating the first pump 803a to supply the eluent from the eluent source 801c to the chromatography device 805 via the first eluent chromatography flow path 820c, and operating the second pump 803b to supply the cleaning liquid from the cleaning source 801b to the first waste liquid container 815 via the second cleaning waste liquid flow path 823b, thereby at least partially simultaneously priming at least the second pump 803b with the cleaning liquid. 960: A step of re - equilibrating the chromatography device 805 by operating the second pump 803b to supply the cleaning liquid from the cleaning source 801b to the chromatography device 805 via the second cleaning chromatography flow path 821b, and operating the first pump 803a to supply the cleaning liquid from the cleaning source 801b to the first waste liquid container 815 via the first cleaning waste liquid flow path 821b, thereby at least partially simultaneously priming at least the first pump 803a with the cleaning liquid. Including.

[0082] During step 950 of performing elution, the eluent from the chromatography device 805 is directed by the lower valve 807 to the fraction collector 808 or the second waste container 809 based on the content of the eluate determined from the signal output from the monitoring unit 806. During all other steps of the above steps, steps 910, 920, 930, 940, 960, the processing liquid output from the chromatography device 805, or typically the mixed processing liquid, is directed by the lower valve 807 to the second waste container 809.

[0083] The duration of priming with the processing liquid in the flow path upstream of the chromatography device 805 in steps 910-960 is determined from the output from the upper monitoring unit 816 or is a predetermined period according to the processing scheme. The durations of washing, equilibration, re-equilibration, and loading of the feed to the chromatography device 805 are preferably based on the output from the monitoring unit 806 (monitoring function). Alternatively, the duration is determined in advance by the processing scheme.

[0084] The preparative chromatography system 800 can be adapted to utilize a plurality of chromatography devices in a manner similar to methods performed in prior art systems.

[0085] One embodiment of a method of utilizing the tandem configuration of the preparative chromatography system 800 is 910’ - Opening the wash source valve 804b:a to connect the wash source 801b and the first pump 803a; - Setting the first upper valve 818a to connect to the common valve 870; - Setting the common valve 870 to connect the first upper valve 818a to the chromatography device 805; - Setting the lower valve 807 to connect to the second waste container 809; - Operating the first pump 803a to provide a cleaning liquid from the cleaning source 801b to the chromatography device 805 and through it; Balancing the chromatography device 805 by; At least partially simultaneously; - Closing the cleaning source valve 804b:b if it is open from the previous cycle; - Opening the supply source valve 804c:b to connect between the supply source 801c and the second pump 803b; - Setting the second upper valve 818b to connect to the first waste container 815; - Operating the second pump 803b to provide a supply liquid from the supply source 801a to the first waste container 815; Priming at least the second pump 803b with the supply liquid by. 920’ - Setting the second upper valve 818b to connect to the common valve 870; - Setting the common valve 870 to connect the second upper valve 818a to the chromatography device 805; - Operating the second pump 803b to provide a supply liquid from the supply source 801a to the chromatography device 805 and through it; Loading the chromatography device 805 with the supply liquid by; At least partially simultaneously; - Setting the first upper valve 818b to connect to the first waste container 815; - Operating the first pump 803a to provide a cleaning liquid from the cleaning source 801b to the first waste container 815; Priming at least the first pump 803a with the cleaning liquid by. 930’ - Setting the first upper valve 818a to connect to the common valve 870; - Setting the common valve 870 to connect the first upper valve 818a to the chromatography device 805; - Operating the first pump 803a to provide a cleaning liquid from the cleaning source 801b to the chromatography device 805, and thereby cleaning the chromatography device 805, at least partially simultaneously, - Closing the supply liquid source valve 804c:b; - Opening the cleaning source valve 804b:b to connect the cleaning source 801b and the second pump 803b; - Setting the second upper valve 818b to connect to the first waste container 815; - Operating the second pump 803b to provide a cleaning liquid from the cleaning source 801b to the first waste container 815, and thereby priming at least the second pump 803b with the cleaning liquid. 940’ - Setting the second upper valve 818b to connect to the common valve 870; - Setting the common valve 870 to connect the second upper valve 818a to the chromatography device 805; - Operating the second pump 803b to provide a cleaning liquid from the cleaning source 801b to the chromatography device 805, and thereby continuing to clean the chromatography device 805, at least partially simultaneously, - Closing the cleaning source valve 804b:a; - Opening the eluent source valve 804c:a to connect the eluent source 801c and the first pump 803a; - Setting the first upper valve 818a to connect to the first waste container 815; - Operating the first pump 803a to provide an eluent from the eluent source 801c to the first waste container 815, and thereby priming at least the first pump 803a with the eluent. 950’ - setting the first upper valve 818a to connect to the common valve 870; - setting the common valve 870 to connect the first upper valve 818a to the chromatography device 805; - operating the first pump 803a to provide an eluent from the eluent source 801c to the chromatography device 805; - based on the output from the monitoring unit indicating the content of the eluate received from the chromatography device 805, setting the lower valve 807 to connect to the fraction collector 808 during the passage of a specific eluate content, and resetting the setting to connect the lower valve 807 to the second waste container 809; by performing elution with the chromatography device 805, at least partially simultaneously, - setting the second upper valve 818b to connect to the first waste container 815; - operating the second pump 803b to provide a cleaning liquid from the cleaning source 801b to the first waste container 815; by priming at least the second pump 803b with the cleaning liquid. 960’ - setting the second upper valve 818b to connect to the common valve 870; - setting the common valve 870 to connect the second upper valve 818a to the chromatography device 805; - operating the second pump 803b to provide a cleaning liquid from the cleaning source 801b to the chromatography device 805; - closing the cleaning source valve 804b:b; by re-equilibrating the chromatography device 805, at least partially simultaneously, - closing the eluent source valve 804c:a; - opening the cleaning source valve 804b:a and connecting the cleaning source 801b and the first pump 803a; - setting the first upper valve 818a to connect to the first waste container 815; - operating the first pump 803a to provide a cleaning liquid from the cleaning source 801b to the first waste container 815; priming at least the first pump 803a with the cleaning liquid. It includes.

[0086] The preparative chromatography system configuration according to the above-described embodiments and the methods that can be utilized using these systems minimize the hold-up volume and shorten the time required for changing from one processing liquid to another. Figures 10a-10b are schematic chromatograms (arbitrary units) of UV absorbance (solid line) and pH (dashed line) versus time during one processing cycle for a) a typical prior art system, and b) a system according to an embodiment described with reference to Figures 2a-2h operated according to the method described with reference to Figures 3a-3h. The processing cycle consists of the stages of loading (1), cleaning (2), elution (3), cleaning / standing cleaning (4), and re-equilibration (5). Shown in Figure 10a is the dispersion period D that surrounds the peak and causes a significant spread. Compared with Figure 10b, the dispersion is significantly reduced, the detection / separation is improved, and the time until one processing cycle is completed is significantly shortened.

[0087] Figure 11 is a chromatogram showing UV absorbance and pH versus time using the preparative chromatography system and method of the embodiment described with reference to Figures 8-10 (solid line) compared to a conventional system (dashed line). It shows how the drawbacks of the conventional system, namely diffusion cleaning buffer transfer, not reaching the appropriate pH quickly enough during cleaning, and slow re-equilibration after the CiP (Cleaning-in-Place stage) cleaning step, all of which are alleviated by the system and method according to the present invention.

[0088] In terms of volume, the system of the present invention saves 10 column volumes per run. In terms of time savings, approximately 0.8 minutes are saved per sub-run, corresponding to 2.6 hours in 200 cycles (which represents the column life), and the execution time is reduced by 15%. See Table 1 below. "Tandem" refers to the preparative chromatography system 800 described with reference to FIGS. 8a - 8b and FIG. 9.

[0089] [Table 1]

[0090] Various embodiments of the present invention can incorporate fiber-based chromatography media and can be optimized for the efficiency of their operation. For example, various embodiments of the present invention can include chromatography components such as one or more Fibro™ / Fibro PrismA™ as commercially available from Cytiva™ Life Sciences.

[0091] Accordingly, various embodiments of the present invention can be provided that have reduced dead volume and / or buffer transfer between each processing step compared to prior art devices / systems. Thus, certain embodiments of the present invention operate faster than known devices / systems and / or have improved theoretical maximum operating efficiency. For example, various embodiments can be provided in which convection is provided to reduce or avoid time delays that occur mainly in diffusion-based devices / systems. Further, various embodiments of the present invention can be provided to have a reduced resin volume compared to known devices / systems. Certain embodiments can additionally or alternatively provide for recycle processing for each of a plurality of capture / elution steps therein (e.g., 200× capture / elution recycle processing can be used).

[0092] For example, in one known chromatography system, the default setting for the delay volume is 8 ml. However, in practice, depending on the mixer used, etc., the delay volume may be 4 - 6 ml. However, by using various embodiments of the present invention, the delay volume (e.g., between the column / injection valve and the chromatography unit such as the separation column) can be advantageously reduced to only 0.5 - 1.0 ml.

[0093] In a known chromatography system incorporating a fiber-based chromatography medium, the membrane volume (mV) therein provides a typical buffer transfer of 10 - 15 mV for a 0.4 ml component (having a 4 ml delay volume). In contrast, various embodiments of the present invention enable such buffer transfer to be reduced to only 3 - 6 mV.

[0094] The above embodiments should be understood as exemplary examples of the systems and methods of the present invention. It will be understood by those skilled in the art that various modifications, combinations, and changes can be made to the above embodiments. In particular, when technically possible, the solutions of different parts in different embodiments can be combined in other configurations.

[0095] [Item 1] A method of performing preparative chromatography by operating a preparative chromatography system (200, 500, 800), wherein the preparative chromatography system (200, 500, 800) is configured to operate with at least a first processing liquid and a second processing liquid, the preparative chromatography system comprising at least one chromatography device (205, 805), a first processing liquid source (201a, 801a), and a second processing liquid source (201b, 801b), a first upper flow path from the first processing liquid source to the chromatography device, a second upper flow path from the second processing liquid source to the chromatography device (205, 805), a first pump (203a, 803a) provided in the first upper flow path and configured to provide the first processing liquid to the chromatography device (205, 805), and a second pump (203b, 803b) provided in the second upper flow path and configured to provide the second processing liquid to the chromatography device (205, 805), the first upper flow path merging with the second upper flow path at a mixing point (221, 821), the mixing point (221, 821) being downstream of the first pump (203a, 803a) and the second pump (203b, 803b) and upstream of the chromatography device (205, 805), the method comprising: - operating the first pump (203a, 803a) to prime the first upper flow path with the first processing liquid, and after the priming step, operating the first pump (203a, 803a) to provide the first processing liquid to the chromatography device (205, 805); and - operating the second pump (203b, 803b) to prime the second upper flow path with the second processing liquid, and after the priming step, operating the second pump (203b, 803b) to provide the second processing liquid to the chromatography device (205, 805). A method comprising the above steps. [Item 2] The preparative chromatography system (200) is configured to operate with at least three processing liquids including a supply liquid, a cleaning liquid, and an eluent. The preparative chromatography system (200) includes a supply source (201a), a cleaning source (201b), and an eluent source (201c), and provides an upper supply flow path (220a) from the supply source (201a) to the chromatography device (205), an upper cleaning flow path (220b) from the cleaning source (201b) to the chromatography device (205), and an upper eluent flow path (220c) from the eluent source (201c) to the chromatography device (205). The method includes - (400:1) priming the upper supply flow path (220a) with the supply liquid provided from the supply source (201a); - (400:2) priming the upper cleaning flow path (220b) with the cleaning liquid provided from the cleaning source (201b); - (400:3) priming the upper eluent flow path (220c) with the eluent provided from the eluent source (201c); and includes a previous stage (400) including The method includes - (410) equilibrating the chromatography device by providing the cleaning liquid from the cleaning source (201b) to the chromatography device (205) via the upper cleaning flow path (220b); - (420) loading the supply liquid into the chromatography device by providing the supply liquid from the cleaning source (201a) to the chromatography device (205) via the upper supply flow path (220a); - (430) cleaning the chromatography device by providing the cleaning liquid from the cleaning source (201b) to the chromatography device (205) via the upper cleaning flow path (220b); - (440) Providing the eluent provided from the eluent source (201c) to the chromatography device (205) via the upper eluent flow path (220c) to perform elution in the chromatography device (205); - (450) Providing the cleaning liquid from the cleaning source (201b) to the chromatography device (205) via the upper cleaning flow path (220b) to re-equilibrate the chromatography device; including a separation step including a processing cycle involving The method according to claim 1. [Claim 3] The preparative chromatography system (200) includes a supply pump (203a) provided in the upper supply flow path (220a) and configured to provide a supply liquid to the chromatography device (205), and an upper supply valve (204a) provided downstream of the supply pump (203a) in the upper supply flow path (220a); a cleaning pump (203b) provided in the upper cleaning flow path (220b) and configured to provide a cleaning liquid to the chromatography device (205), and an upper cleaning valve (204b) provided downstream of the cleaning pump (203b) in the cleaning flow path (220b); an eluent pump (203c) provided in the upper eluent flow path (220c) and configured to provide an eluent to the chromatography device (205), and an upper eluent valve (204c) provided downstream of the eluent pump (203c) in the eluent flow path (220c); a monitoring unit (206) provided downstream of the chromatography device (205), and a lower valve (207) provided downstream of the monitoring unit (206) and configured to connect to either a waste container (209) or a fraction collector (208); and In the method, the previous step (400) - (400:1) Controlling the supply pump (203a) and the upper supply valve (204a) to load the supply liquid into the upper supply flow path (220a) to prime the upper supply flow path (220a), wherein during this supply priming period, the upper cleaning valve (204b) and the upper eluent valve (204c) are closed; - (400:2) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to prime the upper cleaning flow path (220b) by loading the cleaning liquid into the upper cleaning flow path (220b), wherein during this cleaning priming period, the upper supply valve (204a) and the upper eluent valve (204c) are closed; - (400:3) Controlling the eluent pump (203c) and the upper eluent valve (204c) to load the eluent into the upper eluent flow path (220c) to prime the upper eluent flow path (220c), wherein during this eluent priming period, the upper cleaning valve (204b) and the upper supply valve (204a) are closed; including; The active ingredient separation step is as follows: - (410) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to load the cleaning liquid into and through the chromatography device (205) to equilibrate the chromatography device, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper supply valve (204a) and the upper eluent valve (204c) are closed; - (420) Controlling the supply pump (203a) and the upper supply valve (204a) to load the supply liquid into the chromatography device (205) and through it, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper cleaning valve (204b) and the upper eluent valve (204c) are closed; - (430) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to load the cleaning liquid into the chromatography device (205) and through it, thereby cleaning the chromatography device, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper supply valve (204a) and the upper eluent valve (204c) are closed; - (440) Controlling the eluent pump (203c) and the upper eluent valve (204c) to load the eluent into the chromatography device (205) and through it, thereby performing elution in the chromatography device (205), wherein the lower valve (207) is set to either the waste container (209) or the fraction collector (208), and the setting depends on the content of the eluate detected by the monitoring unit (206), and during this step, the upper cleaning valve (204b) and the upper supply valve (204a) are closed; - (450) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to load the cleaning liquid into the chromatography device (205) and through it, thereby re - equilibrating the chromatography device, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper supply valve (204a) and the upper eluent valve (204c) are closed; including The method according to claim 1 or 2. [Item 4] The preparative chromatography system is configured to operate with at least three processing liquids, namely a supply liquid, a cleaning liquid, and an eluent, and the preparative chromatography system - a supply source (201a) connected to a supply pump (203a), a cleaning source (201b) connected to a cleaning pump (203b), and an eluent source (201c) connected to an eluent pump (203c); - A first chromatography device (205a) associated with a first monitoring unit (206a), a second chromatography device (205b) associated with a second monitoring unit (206b), and a third chromatography device (205c) associated with a third monitoring unit (206c), - A first upper supply flow path (220a-1) provided from the supply source (201a) to the first chromatography device (205a), a second upper supply flow path (220a-2) provided from the supply source (201a) to the second chromatography device (205b), and a third upper supply flow path (220a-3) provided from the supply source (201a) to the third chromatography device (205c), wherein the upper supply flow paths are bifurcated downstream of the supply pump (203a), the first, second, and third upper supply flow paths, - A first upper cleaning flow path (220b-1) provided from the cleaning source (201b) to the first chromatography device (205a), a second upper cleaning flow path (220b-2) provided from the cleaning source (201a) to the second chromatography device (205b), and a third upper cleaning flow path (220b-3) provided from the cleaning source (201a) to the third chromatography device (205c), wherein the upper cleaning flow paths are bifurcated downstream of the cleaning pump (203b), the first, second, and third upper cleaning flow paths, - A first upper eluent flow path (220c-1) provided from the eluent source (201c) to the first chromatography device (205a), a second upper eluent flow path (220b-2) provided from the eluent source (201a) to the second chromatography device (205b), and a third upper eluent flow path (220c-3) provided from the eluent source (201a) to the third chromatography device (205c), wherein the upper eluent flow paths are bifurcated downstream of the eluent pump (203c), the first, second, and third upper eluent flow paths, comprising, The method is at least partially executed simultaneously - (710:1) By operating the cleaning pump (203b), one of the first, second, or third chromatography apparatuses (205a; 205b; 205c) is equilibrated or cleaned with the cleaning liquid provided from the cleaning source through one of the first upper cleaning flow paths (220b-1; 220b-2; 220b-3). - (710:2) By operating the supply pump (203a), another one of the first, second, or third chromatography apparatuses (205a; 205b; 205c) is loaded with the supply liquid provided from the supply source (201a) through one of the upper supply flow paths (220a-1; 220a-2; 220a-3). - (710:3) By operating the eluent pump (203c) to provide the eluent from the eluent source (201c) through one of the upper eluent flow paths (220c-1; 220c-2; 220c-3), elution is performed in the remaining part of the first, second, or third chromatography apparatus (205a; 205b; 205c), and the eluate is directed to the waste container or the fraction collector according to the content of the eluate detected by the monitoring unit (206). Including an active fractionation step including The method according to any one of claims 1 to 3. [Claim 5] The method - (700:1) Priming the first upper supply flow path (220a-1) associated with the first chromatography apparatus (205a) with the supply liquid provided from the supply source (201a). Priming the second upper supply flow path (220a-2) associated with the second chromatography apparatus (205b) with the supply liquid provided from the supply source (201a), and priming the third upper supply flow path (220a-3) associated with the third chromatography apparatus (205c) with the supply liquid provided from the supply source (201a), where the supply pump (203a) is operating. - Step of priming the first upper cleaning channel (220b-1) related to the first chromatography device (205a) with the cleaning liquid provided from the cleaning source (201b), step of priming the second upper cleaning channel (220b-2) related to the second chromatography device (205b) with the cleaning liquid provided from the cleaning source (201a), and step of priming the third upper cleaning channel (220a-3) related to the third chromatography device (205c) with the cleaning liquid provided from the cleaning source (201a), wherein the cleaning pump (203b) is operating, and the step; - Step of priming the first upper eluent channel (220c-1) related to the first chromatography device (205a) with the eluent provided from the eluent source (201c), step of priming the second upper eluent channel (220c-2) related to the second chromatography device (205b) with the eluent provided from the eluent source (201c), and step of priming the third upper eluent channel (220c-3) related to the third chromatography device (205c) with the eluent provided from the eluent source (201c), wherein the eluent pump (203c) is operating, and the step; The method according to claim 4, including the previous step (700) involving the above. [Claim 6] The method is - (710:1) Step of equilibrating or cleaning the first chromatography device (205a) with the cleaning liquid provided from the cleaning source (201b) through the first upper cleaning channel (220b-1) by operating the cleaning pump (203b), - (710:2) Step of loading the second chromatography device (205b) with the supply liquid provided from the supply source (201a) through the second upper supply channel (220a-2) by operating the supply pump (203a); - Step of performing elution in the third chromatography device (205c) by operating the eluent pump (203c) to provide eluent from the eluent source (201c) through the third upper eluent flow path (220c-3), wherein the eluate is directed to a waste container or a fraction collector according to the content of the eluate detected by the monitoring unit (206). Or - Step of loading the supply liquid provided from the supply source (201a) through the third upper supply flow path (220a-3) into the third chromatography device (205c) by operating the supply pump (203a). - Step of performing elution in the second chromatography device (205c) by operating the eluent pump (203c) to provide eluent from the eluent source (201c) through the second upper eluent flow path (220c-2), wherein the eluate is directed to a waste container or the fraction collector according to the content of the eluate detected by the monitoring unit (206). Step in which at least one of the above is executed. - Step of equilibrating or cleaning the second chromatography device (205b) with the cleaning liquid provided from the cleaning source (201b) through the second upper cleaning flow path (220b-2) by operating the cleaning pump (203b). - Step of loading the supply liquid provided from the supply source (201a) through the first upper supply flow path (220a-1) into the first chromatography device (205a) by operating the supply pump (203a). - (720:3) By operating the eluent pump (203c) to provide an eluent from the eluent source (201c) through the third upper eluent flow path (220c-3), performing elution in the third chromatography device (205c), wherein the eluate is directed to a waste container or the fraction collector according to the content of the eluate detected by the monitoring unit (206). Or - (720:2’) By operating the supply pump (203a), loading the supply liquid provided from the supply source (201a) through the third upper supply flow path (220a-3) into the third chromatography device (205c). - (720:3’) By operating the eluent pump (203c) to provide an eluent from the eluent source (201c) through the first upper eluent flow path (220c-1), performing elution in the first chromatography device (205a), wherein the eluate is directed to a waste container or the fraction collector according to the content of the eluate detected by the monitoring unit (206). At least one of the above is executed. - (730:1) By operating the cleaning pump (203b), equilibrating or cleaning the third chromatography device (205c) with the cleaning liquid provided from the cleaning source (201b) through the third upper cleaning flow path (220b-3). - (720:2) By operating the supply pump (203a), loading the supply liquid provided from the supply source (201a) through the first upper supply flow path (220a-1) into the first chromatography device (205a). - (720:3) By operating the eluent pump (203c), providing an eluent from the eluent source (201c) through the second upper eluent flow path (220c-2), and performing elution in the second chromatography apparatus (205b), wherein the eluate is directed to a waste container or the fraction collector according to the content of the eluate detected by the monitoring unit (206). Or - (720:2’) By operating the supply pump (203a), loading a supply liquid provided from the supply source (201a) through the second upper supply flow path (220a-2) into the second chromatography apparatus (205b). - (720:3’) By operating the eluent pump (203c), providing an eluent from the eluent source (201c) through the first upper eluent flow path (220c-1), and performing elution in the first chromatography apparatus (205a), wherein the eluate is directed to a waste container or the fraction collector according to the content of the eluate detected by the monitoring unit (206). At least one of the above steps is executed. Including an active fractionation step. The method according to any one of claims 1 to 5. [Claim 7] The fractionation chromatography system - The supply pump (203a) is connected to a first four-way valve (501), the washing pump (203b) is connected to a second four-way valve (502), and the eluent pump (203c) is connected to a third four-way valve (503). - The first four-way valve (501) is connected to a fourth four-way valve (504), a fifth four-way valve (505), and a sixth four-way valve (506). - The second four-way valve (502) is connected to the fourth four-way valve (504), the fifth four-way valve (505), and the sixth four-way valve (506). - The third four-way valve (502) is connected to the fourth four-way valve (504), the fifth four-way valve (505), and the sixth four-way valve (506). - The fourth four-way valve (504) is connected to the first chromatography device (205a), the fifth four-way valve (505) is connected to the second chromatography device (205b), and the sixth four-way valve (506) is connected to the third chromatography device (205c). It is configured according to the following: In the previous step and / or the preparation step: From the supply source (201a) - By setting the first four-way valve (501) to direct the flow towards the fourth four-way valve (504) and setting the fourth four-way valve (504) to direct the flow towards the first chromatography device (205a), the first upper supply flow path (220a-1) is used to supply to the first chromatography device (205a). - By setting the first four-way valve (501) to direct the flow towards the fifth four-way valve (505) and setting the fifth four-way valve (505) to direct the flow towards the second chromatography device (205b), the second upper supply flow path (220a-2) is used to supply to the second chromatography device (205b), and - By setting the first four-way valve (501) to direct the flow towards the sixth four-way valve (506) and setting the sixth four-way valve (506) to direct the flow towards the third chromatography device (205c), the third upper supply flow path (220a-3) is used to supply to the third chromatography device (205c). The supply liquid is provided. From the cleaning source (201b) - By setting the second four-way valve (502) to direct the flow towards the fourth four-way valve (504) and setting the fourth four-way valve (504) to direct the flow towards the first chromatography device (205a), the first upper cleaning flow path (220b-1) is used to supply to the first chromatography device (205a). - By setting the second four-way valve (502) to direct the flow towards the fifth four-way valve (505) and setting the fifth four-way valve (505) to direct the flow towards the second chromatography device (205b), the second upper cleaning flow path (220b-2) is used to supply cleaning liquid to the second chromatography device (205b), and - By setting the second four-way valve (502) to direct the flow towards the sixth four-way valve (506) and setting the sixth four-way valve (506) to direct the flow towards the third chromatography device (205c), the third upper cleaning flow path (220b-3) is used to supply cleaning liquid to the third chromatography device (205c), cleaning liquid is provided, from the eluent source (201c) - By setting the third four-way valve (503) to direct the flow towards the fourth four-way valve (504) and setting the fourth four-way valve (504) to direct the flow towards the first chromatography device (205a), the first upper eluent flow path (220c-1) is used to supply eluent to the first chromatography device (205a), - By setting the third four-way valve (503) to direct the flow towards the fifth four-way valve (505) and setting the fifth four-way valve (505) to direct the flow towards the second chromatography device (205b), the second upper eluent flow path (220c-2) is used to supply eluent to the second chromatography device (205b), and - By setting the third four-way valve (503) to direct the flow towards the sixth four-way valve (506) and setting the sixth four-way valve (506) to direct the flow towards the third chromatography device (205c), the third upper eluent flow path (220c-3) is used to supply eluent to the third chromatography device (205c), eluent is provided, The method according to claim 6. [Claim 8] The preparative chromatography system (800) is configured to operate with at least three process liquids including a supply liquid, a cleaning liquid, and an eluent, and the preparative chromatography system (800) A supply source (801a), a cleaning source (801b), and an eluent source (801c), and a first pump (803a), a second pump (803b), a first waste container (815), and a common valve (870) connected to the chromatography device (805), wherein the preparative chromatography system (800) has a first chromatography flow path (820a, 820b, 820c) that passes the first pump (803a) from at least one of the treatment liquid sources to the common valve (870), and a second chromatography flow path (822a, 822b, 822c) that passes the second pump (803b) from at least one of the treatment liquid sources to the common valve (870), and the first pump (803a), the second pump (803b), the first waste container (815), and the common valve (870) that provide a first waste flow path (821a, 821b, 821c) that passes the first pump (803a) from at least one of the treatment liquid sources to the first waste container (815), and a second waste liquid flow path (823a, 823b, 823c) that passes the second pump (803b) from at least one of the treatment liquid sources to the common valve (870), and comprises In the step of the method, while the first pump (803a) is engaged in providing a treatment liquid to the chromatography device (805), the second pump (803b) is at least partially simultaneously engaged in providing a treatment liquid to the waste container (815) via one of the second waste flow paths (821a, 821b, 821c). The method according to any one of claims 1 to 7. [Claim 9] A predetermined chromatography apparatus sequence defines the order of process fluids passing through the chromatography apparatus (805), and in one step, one of the first pump (803a) and the second pump (803b) provides a process fluid to the chromatography apparatus (805), and at least partially simultaneously, the other of the first pump (803a) and the second pump (803b) primes its associated flow path with a process fluid that orders the next process fluid to be provided to the chromatography apparatus (805) according to the chromatography apparatus. The method according to item 8. [Item 10] - (910) A step of equilibrating the chromatography apparatus (805) by operating the first pump (803a) to provide a cleaning liquid from the cleaning source (801b) to the chromatography apparatus (805) via the first cleaning chromatography flow path (820b), and operating the second pump (803b) to provide a supply liquid from the supply source (801a) to the first waste container (815) via the second supply waste flow path (823a), thereby at least partially simultaneously priming the second pump (803b) with the supply liquid. - (920) A step of loading a supply to the chromatography apparatus (805) by operating the second pump (803b) to provide a supply liquid from the supply source (801a) to the chromatography apparatus (805) via the second supply chromatography flow path (822a), and operating the first pump (803a) to provide a cleaning liquid from the cleaning source (801b) to the first waste container (815) via the first cleaning waste flow path (821b), thereby at least partially simultaneously priming at least the first pump (803a) with the cleaning liquid. - (930) Operating the first pump (803a) to provide a cleaning liquid from the cleaning source (801b) to the chromatography device (805) via the first cleaning chromatography flow path (820b), thereby cleaning the chromatography device (805), and operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the first waste liquid container (815) via the second cleaning waste liquid flow path (823b), thereby at least partially simultaneously priming at least the second pump (803b) with the cleaning liquid. - (940) Continuing to clean the chromatography device (805) by operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the chromatography device (805) via the second cleaning chromatography flow path (821b), and operating the first pump (803a) to provide an eluent liquid from the eluent source (801c) to the first waste container (815) via the first eluent waste disposal flow path (821c), thereby at least partially simultaneously priming at least the first pump (803a) with the eluent liquid. - (950) Executing elution in the chromatography device (805) by operating the first pump (803a) to provide an eluent from the eluent source (801c) to the chromatography device (805) via the first eluent chromatography flow path (820c), and operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the first waste liquid container (815) via the second cleaning waste liquid flow path (823b), thereby at least partially simultaneously priming at least the second pump (803b) with the cleaning liquid. - Operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the chromatography device (805) through the second cleaning chromatography flow path (821b), thereby re-equilibrating the chromatography device (805), wherein the first pump (803a) is operated to provide a cleaning liquid from the cleaning source (801b) to the first waste container (815) through the first cleaning waste flow path (821b), and at least partially simultaneously, at least the first pump (803a) is primed with the cleaning liquid. The method according to claim 8 or 9, comprising. [Claim 11] During the step (950) of performing elution, the eluate from the chromatography device (805) is directed by the lower valve (807) to the fraction collector (808) or the second waste container (809) based on the content of the eluate determined from the signal output from the monitoring unit (806). The method according to claim 10. [Claim 12] The preparative chromatography system (800) includes an upper monitoring unit (816) configured in front of the flow direction to the first waste container (815), and the upper monitoring unit (816) is configured to analyze the content in the first waste flow paths (821a, 821b, 821c) and the second waste flow paths (823a, 823b, 823c). In the step of the method, the duration of at least one of the priming operations is determined based on the output of the measuring unit. The method according to claim 10 or 11. [Claim 13] A preparative chromatography system (200, 500, 800) configured to operate with at least a first processing liquid and a second processing liquid, the preparative chromatography system comprising at least one chromatography device (205, 805), a first processing liquid source (201a, 801a), and a second processing liquid source (201b, 801b), a first upper flow path from the first processing liquid source to the chromatography device, a second upper flow path from the second processing liquid source to the chromatography device (205, 805), a first pump (203a, 803a) provided in the first upper flow path and configured to provide the first processing liquid to the chromatography device (205, 805), and a second pump (203b, 803b) provided in the second upper flow path and configured to provide the second processing liquid to the chromatography device (205, 805), the first upper flow path merging with the second upper flow path at a mixing point (221, 821), the mixing point (221, 821) being downstream of the first pump (203a, 803a) and the second pump (203b, 803b) and upstream of the chromatography device (205, 805), the preparative chromatography system being - In the step of priming the first upper flow path with the first processing liquid that activates the first pump (203a, 803a), and in the steps performed after the priming step, activating the first pump (203a, 803a) to provide the first processing liquid to the chromatography device (205, 805), - In the step of priming the second upper flow path with the second processing liquid, activating the second pump (203b, 803b), and in the steps performed after the priming step, activating the second pump (203b, 803b) to provide the second processing liquid to the chromatography device (205, 805), A preparative chromatography system (200, 500, 800), characterized in that it is configured as described above. [Item 14] The preparative chromatography system (200) is configured to operate with at least three processing fluids, namely a feed solution, a cleaning solution, and an eluent. The preparative chromatography system (200) includes a feed source (201a), a cleaning source (201b), and an eluent source (201c), and provides an upper feed flow path (220a) from the feed source (201a) to the chromatography device (205), an upper cleaning flow path (220b) from the cleaning source (201b) to the chromatography device (205), and an upper eluent flow path (220c) from the eluent source (201c) to the chromatography device (205). The preparative chromatography system (200) is - (400:1) priming the upper feed flow path (220a) with the feed solution provided from the feed source (201a); - (400:2) priming the upper cleaning flow path (220b) with the cleaning solution provided from the cleaning source (201b); - (400:3) priming the upper eluent flow path (220c) with the eluent provided from the eluent source (201c), and is configured to operate the previous stage (400) in this way. The preparative chromatography system (200) is - (410) equilibrating the chromatography device by providing a cleaning solution from the cleaning source (201b) to the chromatography device (205) via the upper cleaning flow path (220b); - (420) loading a feed solution into the chromatography device by providing a feed solution from the feed source (201a) to the chromatography device (205) via the upper feed flow path (220a); - (430) cleaning the chromatography device by providing a cleaning solution from the cleaning source (201b) to the chromatography device (205) via the upper cleaning flow path (220b). - By providing the eluent liquid provided from the eluent source (201c) to the chromatography device (205) through the upper eluent flow path (220c), performing a step of elution in the chromatography device (205); - By providing a cleaning liquid from the cleaning source (201b) to the chromatography device (205) through the upper cleaning flow path (220b), performing a step of re-equilibrating the chromatography device; configured to operate an active fractionation stage including a processing cycle having; the fractionation chromatography system (200) according to claim 13. [Claim 15] a supply pump (203a) provided in the upper supply flow path (220a) and configured to provide a supply liquid to the chromatography device (205), and an upper supply valve (204a) provided downstream of the supply pump (203a) in the upper supply flow path (220a); a cleaning pump (203b) provided in the upper cleaning flow path (220b) and configured to provide a cleaning liquid to the chromatography device (205), and an upper cleaning valve (204b) provided downstream of the cleaning pump (203b) in the cleaning flow path (220b); an eluent pump (203c) provided in the upper eluent flow path (220c) and configured to provide an eluent to the chromatography device (205), and an upper eluent valve (204c) provided downstream of the eluent pump (203c) in the eluent flow path (220c); a monitoring unit (206) provided downstream of the chromatography device (205); a lower valve (207) provided downstream of the monitoring unit (206) and configured to connect to either a waste container (209) or a fraction collector (208); further comprising; the fractionation chromatography system (200) is - (400:1) Controlling the supply pump (203a) and the upper supply valve (204a) to load a supply liquid into the supply channel (220a) to prime the upper supply channel (220a), wherein during this supply priming period, the upper cleaning valve (204b) and the upper eluent valve (204c) are closed, and - (400:2) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to load a cleaning liquid into the cleaning channel (220b) to prime the upper cleaning channel (220b), wherein during this cleaning priming period, the upper supply valve (204a) and the upper eluent valve (204c) are closed, and - (400:3) Controlling the eluent pump (203c) and the upper eluent valve (204c) to load an eluent into the eluent channel (220c) to prime the upper eluent channel (220c), wherein during this eluent priming period, the upper cleaning valve (204b) and the upper supply valve (204a) are closed, and is configured to operate the previous step (400), The preparative chromatography system (200) is - (410) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to load a cleaning liquid into and through the chromatography device (205) to equilibrate the chromatography device, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper supply valve (204a) and the upper eluent valve (204c) are closed, and - (420) Controlling the supply pump (203a) and the upper supply valve (204a) to load a supply liquid into and through the chromatography device (205) to load a supply liquid into the chromatography device, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper cleaning valve (204b) and the upper eluent valve (204c) are closed, and - (430) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to load a cleaning liquid into and through the chromatography device (205) to clean the chromatography device, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper supply valve (204a) and the upper eluent valve (204c) are closed; - (440) Controlling the eluent pump (203c) and the upper eluent agent valve (204c) to load an eluent agent liquid into and through the chromatography device (205) to perform elution in the chromatography device (205), wherein the lower valve (207) is set to either the waste container (209) or the fraction collector (208), the setting depending on the content of the eluate detected by the monitoring unit (206), and during this step, the upper cleaning valve (204b) and the upper supply valve (204a) are closed; - (450) Controlling the cleaning pump (203b) and the upper cleaning valve (204b) to load a cleaning liquid into and through the chromatography device (205) to re - equilibrate the chromatography device, wherein the lower valve (207) is set to the waste container (209), and during this step, the upper supply valve (204a) and the upper eluent valve (204c) are closed; configured to operate an active fractionation stage including a processing cycle having the above; The fractionation chromatography system (200) according to claim 13 or 14. [Claim 16] The fractionation chromatography system (200) is configured to operate with at least three process liquids including a supply liquid, a cleaning liquid, and an eluent liquid, and the fractionation chromatography system (200) is - A supply source (201a) connected to a supply pump (203a), a cleaning source (201b) connected to a cleaning pump (203b), and an eluent source (201c) connected to an eluent pump (203c), - A first chromatography device (205a) associated with a first monitoring unit (206a), a second chromatography device (205b) associated with a second monitoring unit (206b), and a third chromatography device (205c) associated with a third monitoring unit (206c), - A first upper supply flow path (220a-1) provided from the supply source (201a) to the first chromatography device (205a), a second upper supply flow path (220a-2) provided from the supply source (201a) to the second chromatography device (205b), and a third upper supply flow path (220a-3) provided from the supply source (201a) to the third chromatography device (205c), wherein the upper supply flow paths branch downstream of the supply pump (203a), the first, second, and third upper supply flow paths, - A first upper cleaning flow path (220b-1) provided from the cleaning source (201b) to the first chromatography device (205a), a second upper cleaning flow path (220b-2) provided from the cleaning source (201a) to the second chromatography device (205b), and a third upper cleaning flow path (220b-3) provided from the cleaning source (201a) to the third chromatography device (205c), wherein the upper cleaning flow paths branch downstream of the cleaning pump (203b), the first, second, and third upper cleaning flow paths, - The first upper eluent flow path (220c-1) provided from the eluent source (201c) to the first chromatography device (205a), the second upper eluent flow path (220b-2) provided from the eluent source (201a) to the second chromatography device (205b), and the third upper eluent flow path (220c-3) provided from the eluent source (201a) to the third chromatography device (205c), wherein the upper eluent flow paths branch downstream of the eluent pump (203c), the first, second, and third upper eluent flow paths. Comprising The preparative chromatography system (200) - (710:1) By operating the cleaning pump (203b), one of the first, second, or third chromatography devices (205a; 205b; 205c) is equilibrated or cleaned with the cleaning liquid provided from the cleaning source through one of the first upper cleaning flow paths (220b-1; 220b-2; 220b-3). - (710:2) By operating the supply pump (203a), another one of the first, second, or third chromatography devices (205a; 205b; 205c) is loaded with the supply liquid provided from the supply source (201a) through one of the upper supply flow paths (220a-1; 220a-2; 220a-3). - (710:3) By operating the eluent pump (203c) to provide eluent from the eluent source (201c) through one of the upper eluent flow paths (220c-1; 220c-2; 220c-3), elution is performed in the remaining part of the first, second, or third chromatography device (205a; 205b; 205c), and depending on the content of the eluate detected by the monitoring unit (206), the eluate is directed to the waste container or the fraction collector. Configured to operate an active fractionation stage including a processing cycle having The preparative chromatography system (200) according to any one of claims 13 to 15. [Item 17] The fractionation chromatography system (500) is - The supply pump (203a) is connected to the first four-way valve (501), the cleaning pump (203b) is connected to the second four-way valve (502), and the eluent pump (203c) is connected to the third four-way valve (503). - The first four-way valve (501) is connected to the fourth four-way valve (504), the fifth four-way valve (505), and the sixth four-way valve (506). - The second four-way valve (502) is connected to the fourth four-way valve (504), the fifth four-way valve (505), and the sixth four-way valve (506). - The third four-way valve (502) is connected to the fourth four-way valve (504), the fifth four-way valve (505), and the sixth four-way valve (506), and - The fourth four-way valve (504) is connected to the first chromatography device (205a), the fifth four-way valve (505) is connected to the second chromatography device (205b), and the sixth four-way valve (506) is connected to the third chromatography device (205c). is configured according to The fractionation chromatography system (500) is The supply liquid is from the supply source (201a) - By setting the first four-way valve (501) to direct the flow toward the fourth four-way valve (504) and setting the fourth four-way valve (504) to direct the flow toward the first chromatography device (205a), to the first chromatography device (205a) via the first upper supply flow path (220a-1), - By setting the first four-way valve (501) to direct the flow toward the fifth four-way valve (505) and setting the fifth four-way valve (505) to direct the flow toward the second chromatography device (205b), to the second chromatography device (205b) via the second upper supply flow path (220a-2), and - Set the first four-way valve (501) to direct the flow towards the sixth four-way valve (506), and set the sixth four-way valve (506) to direct the flow towards the third chromatography device (205c). Thus, the third upper supply flow path (220a-3) is used to supply to the third chromatography device (205c) provided with the cleaning liquid from the cleaning source (201b) - Set the second four-way valve (502) to direct the flow towards the fourth four-way valve (504), and set the fourth four-way valve (504) to direct the flow towards the first chromatography device (205a). Thus, the first upper cleaning flow path (220b-1) is used to supply to the first chromatography device (205a) - Set the second four-way valve (502) to direct the flow towards the fifth four-way valve (505), and set the fifth four-way valve (505) to direct the flow towards the second chromatography device (205b). Thus, the second upper cleaning flow path (220b-2) is used to supply to the second chromatography device (205b), and - Set the second four-way valve (502) to direct the flow towards the sixth four-way valve (506), and set the sixth four-way valve (506) to direct the flow towards the third chromatography device (205c). Thus, the third upper cleaning flow path (220b-3) is used to supply to the third chromatography device (205c) provided with the eluent from the eluent source (201c) - Set the third four-way valve (503) to direct the flow towards the fourth four-way valve (504), and set the fourth four-way valve (504) to direct the flow towards the first chromatography device (205a). Thus, the first upper eluent flow path (220c-1) is used to supply to the first chromatography device (205a) - Set the third four-way valve (503) to direct the flow towards the fifth four-way valve (505), and set the fifth four-way valve (505) to direct the flow towards the second chromatography device (205b). Thus, the second upper eluent flow path (220c-2) is used to supply to the second chromatography device (205b), and - By setting the third four-way valve (503) to direct the flow toward the sixth four-way valve (506) and setting the sixth four-way valve (506) to direct the flow toward the third chromatography device (205c), the third chromatography device (205c) is connected via the third upper eluent flow path (220c-3). It is configured to be provided as follows. The preparative chromatography system (500) according to any one of claims 13 to 16. [Claim 18] The preparative chromatography system (800) is configured to operate with at least three processing liquids, namely a supply liquid, a cleaning liquid, and an eluent. The preparative chromatography system (800) - A supply source (801a), a cleaning source (801b), and an eluent source (801c); - A first pump (803a), a second pump (803b), a first waste container (815), and a common valve (870) connected to the chromatography device (805). The preparative chromatography system (800) - A first chromatography flow path (820a, 820b, 820c) that passes the first pump (803a) from at least one of the processing liquid sources to the common valve (870), and a second chromatography flow path (822a, 822b, 822c) that passes the second pump (803b) from at least one of the processing liquid sources to the common valve (870); - A first waste flow path (821a, 821b, 821c) that passes the first pump (803a) from at least one of the treatment liquid sources to the first waste container (815), and a second waste liquid flow path (823a, 823b, 823c) that passes the second pump (803b) from at least one of the treatment liquid sources to the common valve (870). During the treatment cycle, the preparative chromatography system (200) is configured while the first pump (803a) is engaged in providing the treatment liquid to the chromatography device (805), and the second pump (803b) is at least partially simultaneously engaged in providing the treatment liquid to the waste container (815) via one of the second waste flow paths (821a, 821b, 821c). The preparative chromatography system (800) according to any one of claims 13 to 17. [Claim 19] A predetermined chromatography device sequence defines the order of the treatment liquid passing through the chromatography device (805). In one step, one of the first pump (803a) and the second pump (803b) provides the treatment liquid to the chromatography device (805), and at least partially simultaneously, the other of the first pump (803a) and the second pump (803b) primes the associated flow path with the treatment liquid that orders the next treatment liquid provided to the chromatography device (805) according to the chromatography device. The preparative chromatography system (800) according to any one of claims 13 to 18. [Claim 20] The preparative chromatography system (800) is - (910) Operating the first pump (803a) to provide a cleaning liquid from the cleaning source (801b) to the chromatography device (805) via the first cleaning chromatography flow path (820b), thereby equilibrating the chromatography device (805), and operating the second pump (803b) to provide a supply liquid from the supply source (801a) to the first waste container (815) via the second supply waste flow path (823a), thereby at least partially simultaneously priming the second pump (803b) with the supply liquid. - (920) Operating the second pump (803b) to provide a supply liquid from the supply source (801a) to the chromatography device (805) via the second supply chromatography flow path (822a), thereby loading a supply onto the chromatography device (805), and operating the first pump (803a) to provide a cleaning liquid from the cleaning source (801b) to the first waste container (815) via the first cleaning waste flow path (821b), thereby at least partially simultaneously priming at least the first pump (803a) with the cleaning liquid. - (930) Operating the first pump (803a) to provide a cleaning liquid from the cleaning source (801b) to the chromatography device (805) via the first cleaning chromatography flow path (820b), thereby cleaning the chromatography device (805), and operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the first waste liquid container (815) via the second cleaning waste liquid flow path (823b), thereby at least partially simultaneously priming at least the second pump (803b) with the cleaning liquid. - (940) Operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the chromatography device (805) through the second cleaning chromatography flow path (821b), thereby continuing to clean the chromatography device (805), and operating the first pump (803a) to provide an eluent liquid from the eluent source (801c) to the first waste container (815) through the first eluent waste flow path (821c), thereby at least partially priming at least the first pump (803a) with the eluent liquid simultaneously. - (950) Operating the first pump (803a) to provide an eluent from the eluent source (801c) to the chromatography device (805) through the first eluent chromatography flow path (820c), thereby performing elution in the chromatography device (805), and operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the first waste liquid container (815) through the second cleaning waste liquid flow path (823b), thereby at least partially priming at least the second pump (803b) with the cleaning liquid simultaneously. - (960) Operating the second pump (803b) to provide a cleaning liquid from the cleaning source (801b) to the chromatography device (805) through the second cleaning chromatography flow path (821b), thereby re - equilibrating the chromatography device (805), and operating the first pump (803a) to provide a cleaning liquid from the cleaning source (801b) to the first waste container (815) through the first cleaning waste flow path (821b), thereby at least partially priming at least the first pump (803a) with the cleaning liquid simultaneously. configured to operate by performing the above. The preparative chromatography system (800) according to any one of claims 13 to 19. [Claim 21] During the step (950) of performing elution, the eluate from the chromatography apparatus (805) is directed by the lower valve (807) to the fraction collector (808) or the second waste container (809) based on the content of the eluate determined from the signal output from the monitoring unit (806), the preparative chromatography system (800) according to any one of claims 13 to 20. [Claim 22] The preparative chromatography system (800) includes an upper monitoring unit (816) configured in front of the flow direction to the first waste container (815), and the upper monitoring unit (816) is configured to analyze the content in the first waste flow path (821a, 821b, 821c) and the second waste flow path (823a, 823b, 823c), the preparative chromatography system (800) according to any one of claims 13 to 21. [Claim 23] A preparative chromatography system (200, 500, 800) incorporating at least one fiber-based chromatography medium and / or at least one membrane-based medium and configured to provide at least one convective fluid flow therein. [Claim 24] The preparative chromatography system (200, 500, 800) according to claim 23, configured to provide a delay volume / dead volume / hold-up volume of less than 4 ml between two components. [Claim 25] The preparative chromatography system (200, 500, 800) according to claim 24, configured to provide a delay volume / dead volume / hold-up volume of 0.5 ml to 4 ml between the two components. [Claim 26] The preparative chromatography system (200, 500, 800) according to claim 25, configured to provide a delay volume / dead volume / hold-up volume of 0.5 ml to 1.0 ml between the two components. [Claim 27] The preparative chromatography system (200, 500, 800) according to any one of claims 23 to 26, wherein the two components include a column injection valve and a separation column. [Claim 28] The preparative chromatography system (200, 500, 800) according to any one of claims 23 to 27, wherein at least one of the fiber-based chromatography component or the membrane-based component is configured to have a buffer transfer of less than 10 membrane volumes (mV) therein. [Claim 29] The preparative chromatography system (200, 500, 800) according to claim 28, wherein the buffer transfer is 3 to 10 mV. [Claim 30] The preparative chromatography system (200, 500, 800) according to claim 29, wherein the buffer transfer is 3 to 6 mV.

Explanation of Reference Numerals

[0096] 100, 200 Preparative chromatography system 101a, 201a Source 101b, 201c Eluent source 102b, 201b Cleaning solution source 103 First valve 111 First common pipe device 104 Common pump 112 Second common pipe device 105, 205 Chromatography device 106 Monitoring device 107 Lower valve 108 Fraction collector 109 Waste container 110 First common piping configuration 203a Supply pump 204a Upper supply valve 210a First supply pipe device 211a Second supply pipe device 212a Third supply pipe device

Claims

1. A preparative chromatography system (200, 500, 800) incorporating at least one fiber-based chromatography medium and / or at least one membrane-based medium and configured to provide at least one convective fluid flow therein.

2. The preparative chromatography system (200, 500, 800) according to claim 1, configured to provide a delay volume / dead volume / hold-up volume of less than 4 ml between two components.

3. The preparative chromatography system (200, 500, 800) according to claim 2, configured to provide a delay volume / dead volume / hold-up volume of 0.5 ml to 4 ml between the two components.

4. The preparative chromatography system (200, 500, 800) according to claim 3, configured to provide a delay volume / dead volume / hold-up volume of 0.5 ml to 1.0 ml between the two components.

5. The preparative chromatography system (200, 500, 800) according to any one of claims 1 to 4, wherein the two components include a column injection valve and a separation column.

6. The preparative chromatography system (200, 500, 800) according to any one of claims 1 to 5, wherein at least one of the fiber-based chromatography component or the membrane-based component is configured to have a buffer transfer of less than 10 membrane volumes (mV) therein.

7. The preparative chromatography system (200, 500, 800) according to claim 6, wherein the buffer transfer is 3 to 10 mV.

8. The preparative chromatography system (200, 500, 800) according to claim 7, wherein the buffer transfer is 3 to 6 mV.

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